February 15-18, 2009 Washington State Convention Center Seattle ...

February 15-18, 2009
Washington State
Convention Center
Seattle, Washington
THE NATIONAL CONFERENCE & EXPOSITION OF
Associate Sponsors
Health Sponsor
AFIA - Aquaculture Committee Global Aquaculture Alliance
American Tilapia Association International Association of Aquaculture
American Veterinary Medical Association Economics and Management
Aquacultural Engineering Society Latin American Chapter WAS
Aquaculture Association of Canada Striped Bass Growers Association
Catfish Farmers of America US Shrimp Farming Association
Fish Culture Section, AFS US Trout Farmers Association
World Aquatic Veterinary Medical Association

PREFACE
Welcome to Aquaculture America 2009 in the impressive Seattle Convention Center. We last met in this facility in
1997 and those of you who attended that meeting will notice that the facility has undergone a significant remodel
and been nicely upgraded since our previous visit. As a result, all of our meetings and activities will occur on a
single floor this year, making it easier to move from one session to another to hear the presentations you find most
valuable.
Aquaculture has also undergone a kind of remodelling since the last meeting in Seattle, with new challenges, new
opportunities, and new sensibilities emerging simultaneously. However, one thing has not changed. Aquaculture
flourishes because it provides valuable products to the consumer. Accordingly we decided not to use a catchy
slogan for the meeting this year but we do have a theme, namely “Healthy aquaculture products for the consumer.”
Stories regularly appear in the media that describe high contaminant loads in seafood products, suggesting that
consumers are risking their health if they consume seafood. Quite the opposite is true, as our keynote speaker,
Dr. Jay Wheaton will explain in his Plenary Presentation “Fishy Business: Aquaculture, Omega-3 fats and Health.”
Seafood products contain an array of nutrients that promote good health, and the benefits of consuming seafood
outweigh health risks by a factor of 100 to 1, according to reliable estimates by health professionals. Aquaculture
products have a potential advantage over seafood products from capture fisheries because the inputs can be
controlled. Thus, the nutritional benefits of farmed fish products to the consumer can be enhanced and contaminant
levels can be reduced simply by altering the feed used to rear fish. In addition to our Plenary Presentation, there
will be a number of technical presentations at this meeting that illustrate this potential.
Seattle is a seafood-lovers paradise, not only because of its rich history as the center of the seafood industry in the
Northern Pacific, but also because of the influence of diverse Asian communities. You will find some interesting
seafood dishes in Seattle based on specific cultures but also Asian fusion dishes that blend the gastronomic influences
of Asian cultures. Take a free bus ride to Uwajimaya, an enormous, modern supermarket at the southern end of
downtown Seattle, to experience the dynamic and diverse Asian influences that gives Seattle its unique flavor.
Consume some seafood and boost your health. When you are full, remember wireless internet and excellent coffee
are always close by in Seattle.
Dr. Ronald W. Hardy, Conference Chair
Dr. Wendy Sealey, Technical Program Chair
Betsy Hart, Industry Program Chair
2

TABLE OF CONTENTS
Preface..........................................................................2
aquaculture.america.2009.abstracts........................5
abstract.addendum..................................................386
To find abstracts for a specific author or subject, use
the pdf search features built into Adobe Acrobat.
DISCLAIMER
The World Aquaculture Society prints abstracts in this Abstract Book exactly as they are submitted
without editing or confirmation of material contained in the abstract. The World Aquaculture Society has
no responsibility for the information contained in the abstracts. The World Aquaculture Society is not
responsible for authors or contact information contained in the abstracts.
The World Aquaculture Society does not have any liability for problems or damages caused by the use of
the information in the abstracts published in the Abstract Book. Any one using these abstracts needs to
verify the information in the abstracts on their own before utilizing such information and will have full
liability for results of using such information.
3

ABSTRACTS
4

STUDY On MORTALITY EGG Of Acipenser persicus DURING INCUBATION
Abedi .M., Ph.D
Faculty member and Head of Mazandaran Azad Higher Education Institute
P.O.Box:19735-184
Tehran, Iran
Drmabedi@yahoo.com
In the farms of artificial in reproduction and fostering, considering that the perish percentage of egg, has an important aspect
in creating good order in affairs and thrift in power of labour and investment; therefore in this research, take action to appoint
perish percentage egg of Acipenser persicus.
For this work after the artificial reproduction, spilling the eggs in the incubators, in 3 stages:
1- stage of the second division of mitosis
2- stage of final gastrula
3- the stage before division
Then, sampling from the eggs and after fixing them in %10 formalin, they studied under the loop, and healthy and unhealthy
eggs were divided. The results of the perish percentage of eggs from Acipenser persicus , in the mentioned stages, were %30 ,
%34 and %40 respectively, that of course these numbers are under the impression of several factors and for this reason , they
are different , in different conditions . According of this result, we can appoint that the eggs incubation is economical or not?
And is it better to evacuate incubators from the eggs with high perish percentage?

ADULT RED SWAMP CRAYFISH (PROCAMBARUS CLARKII): EFFECTS OF SELENIUM
ENRICHED DIET
Maria Cesarina Abete*, Antonia Concetta Elia, Ilaria Giorgi, A.J. Martin Dörr, Stefania Squadrone, Renata Tarasco,
Maria I. Taticchi and Marino Prearo
State Veterinary Institute
Via Bologna
148 - 10154 Turin –Italy
Selenium (Se) as selenocysteine, is a component of selenoproteins, it has an important enzymatic function in animals because
essential for the proper functioning of the antioxidant enzyme glutathione peroxidase. Many studies on selenium have been
conducted in order to investigate dietary requirements, elimination and uptake rates from water and diet sources in organisms
and toxic levels in water. To our knowledge few data are reported for P. clarkii fed with diets enriched in selenium (Elia
et al., 2007; Dörr et al., 2008). The aim of the present study was to evaluate the effect of selenium on the enzymatic activities
of catalase (CAT), glutathione peroxidase (Se-GPx), and glutathione S-transferase (GST) in hepatopancreas of adult P. clarkii
for both sexes, when treated for 15 days with different diets and the Se accumulation in this tissue.
P. clarkii were fed ad libitum with two diets (0.30 and 1.21 mg/kg Se). After 15 days treatment at specific Se concentrations, 10
specimens from each experimental and control tank were transferred immediately live to the laboratory. For the whole sample
hepatopancreas were removed and immediately stored at –80 °C. The determination of selenium was carried out by atomic
absorption spectrometry. The quantification limit (LOQ) is 0.01 ppm (mg/kg). The same samples (0.5 g) were analyzed for
enzyme activities (CAT, Se-GPx and GST) as reported in the literature.
The concentration of selenium in hepatopancreas was 0.47 ppm in treated females and 0.31 ppm in the control, while in treated
males selenium was 0.55 ppm versus 0.37 ppm in control. Crayfish showed a depletion of CAT activity in females and was
about 70% lower compared to the respective untreated samples. An apposite trend was displayed by Se-GPx, lowered only
in treated males (about 40%), compared to the untreated ones. The Se-GPx and CAT are the two enzymes responsible for the
detoxification of hydrogen peroxide (H 2 O 2 ) in biological systems. The results obtained in the present research indicate that both
sexes of P. clarkii responded differently under a high selenium treatment.
Therefore, the marked low activities of CAT in females and GPx in males might be the result of a compromised oxidative
stress status with a consequent decline of the protection toward the prooxidant effect of Se in both sexes. Most probably the
reduction of these activities could be correlated with the different regulation of enzymes expression between sexes under Se
treatment. GST enzyme, which has the role to detoxify from electrophylic compounds, was not affected by the high selenium
diet treatment. Even if higher activity was recorded in both treated sexes, it was not statistically different compared to their
own controls. Therefore, GST activity was not a sensible marker to detect the Se-prooxidant effect at this experimental condition.
The antioxidant response under selenium exposure might be largely variable depending on species, experimental times,
selenium species and concentrations. In conclusion, these preliminary results indicate that males and females of adult P. clarkii
show a different susceptibility to Se-dietary treatment. More data will be necessary to make more clear the effect of selenium
on these antioxidant enzymes in crayfish.

HEAVY METALS IN THE BIVALVE MOLLUSC FROM NORTH-EAST ITALY
Daniela Marchis, Maria Cesarina Abete*, Monica Caffara, Ilaria Giorgi, Renata Tarasco, Maria Letizia Fioravanti
and Marino Prearo
State Veterinary Institute
Via Bologna
148 - 10154 Turin – Italy
Bivalve molluscs are supposed to be very good pollution markers, as they can tolerate a wide range of environmental factors
and can concentrate some chemical contaminants, as heavy metals. From February to May 2006, periodic samplings of Tapes
philippinarum, in Goro-Porto Garibaldi area, near Ferrara, and of Chamelea gallina, along the coast nearby Rimini (North-East
Italy, Emilia Romagna region) were carried out. 240 C. gallina individuals in 4 different samplings and 540 T. philippinarum
individuals in 9 different samplings were collected.
The aim of this study was to evaluate the amount of some heavy metals which are ruled by Regulations EC 1881/2006 and EC
629/2008 (Cadmium, Lead and Mercury). Moreover, the Chromium level was also detected, as this trace element, which at the
moment has no maximum residue limit in food, appears to be widespread in the environment. Following microwave digestion,
all the samples were analyzed for Cadmium, Lead and Chromium by atomic absorption spectrometry with Zeeman background
corrector. Limits of quantification (LOQ) were 0.01 mg/kg for Cadmium, 0.04 mg/kg for Lead and 0.05 for Chromium. Mercury
was detected using Direct Mercury Analyzer (TDA-AAS) and LOQ was 0.07 mg/kg. C. gallina samples showed a Cadmium
mean concentration of 0.014 mg/kg (SD 0.013), a Chromium mean concentration of 0.4 mg/kg (SD 0.0028), and a Lead mean
level of 0.07 mg/kg (SD 0.021). In all samples Mercury concentration was below the LOQ.
T. philippinarum individuals showed to be contaminated as follow: Cadmium mean concentration 0.007 mg/kg (SD 0.003);
Chromium mean concentration 1.26 mg/kg (SD 0.65), and Lead mean level 0.145 mg/kg (SD 0.064). Mercury appeared to
show a mean concentration above the LOQ (0.11 mg/kg) in one pool only, whilst in all the other samples it was below the LOQ
value.
All samples were compliant to the EU legislation, as all concentrations resulted below the maximum residue limits. Therefore,
these clams meet the requirements for consumers. Nevertheless, as Chromium contamination appears to be constant in both
species, further investigations on water and on other aquatic organisms should be carried out.

PARTIAL AND FULL REPLACEMENT OF MENHADEN FISHMEAL PROTEIN BY
SOYBEAN MEAL PROTEIN SUPPLEMENTED WITH OR WITHOUT AMINO ACIDS AND
ATTRACTANTS IN THE DIETS OF JUVENILE BLACK SEA BASS Centropristis striata
Md. Shah Alam*, Wade O. Watanabe, Katherine B. Sullivan and Walker D. Wright-Moore
University of North Carolina Wilmington
Center for Marine Science
601, S. College Road
Wilmington, NC, 28403-5927 USA
alamm@uncw.edu
A series of experiments were conducted to test the effect of varying dietary levels of solvent-extracted soybean meal supplemented
with or without amino acids and attractants in the diets of juvenile black sea bass. In experiment 1, seven diets were
formulated replacing menhaden fishmeal protein (FP) by soybean meal protein (SP) at 0, 10, 20, 30, 40, 50, and 60% without
supplementing amino acids but adding 1% attractants. In experiment 2, six diets were formulated replacing FP by SP at 0, 60,
70, 80, 90 and 100% without supplementing amino acids but adding 1% attractants. Experiment 3 and 4 were designed the
same as experiments 1 and 2, respectively, but without attractants. Experiment 5 was designed as experiment 4, but supplementing
coated L-methionine and L-lysine. In each experiment, diets were formulated to have about the same protein (44%
crude protein) and lipid level (10% lipid). All diets contained 7.5% squid meal and 5% krill meal. Each test diet was fed to
triplicate groups of juvenile black sea bass held at 15 fish per 74-L recirculating sea water tank for 6 weeks in experiments 1, 3
and 4, 5 and 10 weeks in experiment 2. The fish were fed two times a day up to apparent satiation.
After the feeding trials, survival was more than 85% in all experiments, and there were no significant differences in survival
among treatments in all five experiments. In experiment 1, growth results showed no significant differences (P

THE POTENTIAL OF “LOST CROPS” IN AQUAFEED, THEIR TOXINS AND METHODS
FOR ENHANCING THEIR NUTRITIVE VALUE
Alegbeleye, W.O.*; George, F.O.A.; Obasa, S.O and Olude, O.O.
University of Agriculture
P.M.B., 2240
Abeokuta, Nigeria
segunalgbeleye@yahoo.com
This paper reviews the nutritive potential of “lost crops” as dietary protein sources in aquafeeds and reports studies on the enhancement
of their inherent composition. The crops include oilseeds (castor oil seed, Ricinus communis, Moringa oleifera and
wing bean), legumes (Lima bean, Albizia gygia, jack bean (Canavalia ensiformes)a, Pterocarpus santalinoides and Mucuna
uriens).
These plants have inherent antinutritional factors that limit their inclusion in aquafeeds, including protease inhibitors, hemaglutinnins,
allergins, toxic amino-acids (mimosine), glycocides (goitrogens, cyanogens, saponnins, phytoestrogens), phenols
(gossypol and tannins), phytic acids and poisonous fatty acids. Cheap and adaptive processing techniques (oil extraction, thermal
methods and fermentation) with potentials for inactivating and improving the nutritive value of these plant products were
examined and discussed.

10
LABORATORY CULTURE OF THE DIATOM Chaetoceros gracilis USING EFFLUENT FROM
AN INTENSIVE MARINE RECIRCULATING AQUACULTURE SYSTEM FOR BLACK SEA
BASS Centropristis striatus
Dustin M. Allen* and Wade O. Watanabe
University of North Carolina Wilmington
Center for Marine Science
601 S. College Rd.
Wilmington, NC 28403-5927 USA
DMA5903@uncw.edu
Concerns of effluent management, land availability and access to water resources for marine finfish production have increased
attention to recirculating aquaculture systems (RAS), which conserve water by reusing greater than 90% of its total daily water
volume, but produce an effluent that is highly concentrated. The objective of this study was to evaluate the effluent from a
marine RAS for black sea bass (BSB, Centropristis striatus) as a nutritive base for production of the marine diatom, Chaetoceros
gracilis, a valuable species that is commonly used for the culture of marine bivalves. An experiment was conducted to
evaluate growth of C. gracilis on RAS effluent in 2-L flasks under controlled laboratory conditions (26.3 ± 1.5 ◦ C and 24 L :
0 D). Four treatment media were compared: RAS effluent with silica added, RAS effluent without silica, Guillards f/2 media,
and a seawater control (32 g/L). Media was filtered, sterilized, and then inoculated with C. gracilis at 1 x 10 6 cells/mL. RAS
effluent was filtered (0.45 μm) before use in the experiment to simulate the incorporation of geotube technology in the water
renovation process. Geotubes are used in wastewater treatment to remove solids by passing the effluent through a geotextile
fabric bag containing a polymer to accelerate flocculation.
Cell density (cells/mL) in the control treatment remained near initial levels, fluctuating between 7.11 x 10 5 and 1.21 x 10 6 from
d 0 to d 9. The f/2 media produced better growth, with cell densities increasing steadily to a plateau of 3.42 x 10 6 by d 7. The
effluent without silica produced faster growth than the f/2, with cell densities reaching 4.42 x 10 6 by d 6. The effluent with
silica added produced similar growth to effluent without silica through d 6, but cell densities plateaued at a higher level of 4.98
x 10 6 by d 7. Increasing cell densities were associated with a reduction in nitrogen (N) levels in the culture media. In the effluent
plus silica treatment, mean (sd) nutrient concentrations (mg/L) of TN, NO 3 /NO 2 , PO 4 and TP declined from initial levels
of 14.1 (0.59), 7.43 (0.22), 0.47 (0.03), and 0.81 (0.08), respectively, to 8.13 (0.48), 0.13 (0.08), 0.00 (0.00) and 0.00 (0.00)
by d 8. NH 4 oscillated between 0.61 (0.15) and 1.03 (0.12). Nondetectable levels of phosphorus in the media by d 3 strongly
suggested a phosphate-based limitation to cell growth. These results showed that effluent from an intensive marine RAS for
BSB was an effective nutrient base for growth of C. gracilis under laboratory conditions. These data will be used to scale up
C. gracilis culture to larger bioreactors and to develop techniques for integrated marine finfish-microalgae-shellfish polyculture
as a means of reducing the nutrient loading associated with fish culture and of producing bivalves for habitat restoration and
mitigation projects.

DIETARY CARBOHYDRATE LEVEL AFFECTS EXPRESSION OF HEPATIC GLUCOSE
TRANSPORTERS IN RAINBOW TROUT (Oncorhynchus mykiss): IS RAINBOW TROUT
GLUT2 DIFFERENT FROM MAMMALIAN GLUT2?
Jon J. Amberg*, Gordon K. Murdoch, Barrie D. Robison, Madison S. Powell, Kenneth J. Rodnick, Rodney A. Hill
and Ronald W. Hardy
Department of Forestry and Natural Resources
Purdue University
West Lafayette, IN 47907 USA
jamberg@vandals.uidaho.edu
Little is known about the hepatic facultative glucose transporters (GLUT1 and GLUT2) in rainbow trout (Oncorhynchus
mykiss). Both, GLUT1 and GLUT2 are bi-directional transporters and are transcriptionally regulated by glucose. In mammals,
GLUT1 has been referred to as the basal glucose transport and GLUT2 senses blood glucose levels and then transports glucose
into hepatocytes and β-cells, which leads to the release of insulin. Thus, these transporters are significant factors in regulating
blood glucose levels in mammals but little is known about their role in fish. Rainbow trout are generally considered intolerant
to high levels of dietary carbohydrates and are inefficient at regulating blood glucose levels. This study used quantitative realtime
PCR (qRT-PCR) to evaluate GLUT1 and GLUT2 expression in response to different dietary carbohydrate levels as well as
using an in silico approach to compare amino acid sequences of GLUT2 proteins with other fish and mammalian species.
Rainbow trout were fed diets containing 0, 15, 25 or 35% starch for 12 weeks. Expression patterns of GLUT1 and GLUT2 in
fish were compared between dietary treatment groups using real-time PCR. To compare amino acid sequences of the GLUT2
proteins, we used software packages to identify putative transmembrane domains, glycosylation and phosphorylation sites
and compared the known functional residues and their locations in the amino acid sequences of mammalian and other piscine
GLUT2 with the rainbow trout GLUT2 sequence.
GLUT1 mRNA content was not affected by dietary carbohydrate. However, GLUT2 was down-regulated in fish fed the 25 or
35% carbohydrate diets. These diets resulted in significantly increased plasma glucose levels and high liver glycogen content
in the fish. Thus, when both plasma glucose levels and liver glycogen content are elevated, the rainbow trout liver decreases
GLUT2 expression that would minimize the uptake of glucose by the liver. This differs from the expression pattern of mammalian
GLUT2. Mammalian hepatic GLUT2 expression correlates with blood glucose levels, which allows for increased
hepatic glucose uptake and increased utilization of glucose as an energy source when blood glucose levels increase. These
results suggest there may be functional differences of rainbow trout GLUT2 than mammalian GLUT2. Preliminary in silico
analysis indicates that differences between the GLUT2 proteins may help to explain the rainbow trout’s intolerance to high
dietary carbohydrates.
11

12
FACTORS THAT INFLUENCE PUBLIC PERCEPTIONS OF AQUACULTURE AND
CONSUMPTION OF FARM-RAISED SEAFOOD IN THE PACIFIC NORTHWEST
Shannon M. Amberg* and Troy E. Hall
Department of Forestry and Natural Resources
Purdue University
195 Marsteller Street
West Lafayette, IN 47907-2033 USA
ambergs@purdue.edu
Aquaculture is one of many rapidly developing food technologies that hold promise to provide high quality, reasonably priced
foods for a growing world population. Like many recent food technologies, however, aquaculture has received mixed attention
in the media (Amberg & Hall, 2008). Little is known about what drives public attitudes toward aquaculture, specifically
those factors that lead to positive or negative attitudes or influence purchase behavior. This study investigates consumers’
attitudes toward aquaculture and the factors that influence perceptions of farm-raised seafood and as well as their purchasing
behavior. Researchers conducted a mail survey that was administered to a random selection of residents (≥18 years) in Idaho
and Washington (n=1215). Consumers were asked about a number of topics including their general beliefs about seafood, their
seafood eating habits, factors that are important in their choices about seafood, frequency of consuming various farm-raised
products, their beliefs about aquaculture, awareness of issues and positive or negative media attention regarding aquaculture,
sources of information about seafood, and appropriate socio-demographic questions. Results show that many survey respondents
find seafood to be a healthy food option and the majority (70%) indicated that they eat fish or seafood at least 2-3 times
per month. Results also show that, on average, respondents indicated a preference for wild seafood over farm-raised products.
For example, 26% said they consumer farm-raised salmon at least once per month, while 50% reported selecting wild salmon.
This preference is clarified in consumers’ responses regarding their beliefs about aquaculture, in that respondents moderately
agreed that wild fish are healthier and have less contamination because the live in a natural environment and eat a natural diet,
whereas they believe that farm-raised fish usually have color or hormones added to them and the crowded conditions on farms
is unhealthy. The impact of media information on consumers’ seafood choices and the correlation between consumers’ awareness
of media reports and general issues regarding aquaculture and their beliefs is also discussed.

EFFICIENT AND RELIABLE PROTOCOLS FOR THE PRODUCTION OF LIVE FEEDS FOR
LARVAL FLORIDA POMPANO
Meghan L. Anderson*, Charles R. Weirich and Fernando G. Cavalin
USDA Agricultural Research Service and Center for Aquaculture and Stock Enhancement
Harbor Branch Oceanographic Institute
Florida Atlantic University
5600 US Highway 1 North
Ft. Pierce, FL 34946 USA
mander62@hboi.fau.edu
As is the case with most marine finfish species, production of live feed organisms represents the majority of time and labor
associated with larviculture operations. As a byproduct of establishing a reproduction and larviculture research program at
our facility, procedures for the production and enrichment of rotifers and Artemia for the successful rearing of larval Florida
pompano have been improved and streamlined. Our established live feeds culture protocols are efficient, reliable, and easily
support both research and commercial-scale larviculture of this species.
Rotifers are batch cultured using a series of four fiberglass tanks containing 125 L of water. Water temperature and salinity are
maintained at 27-28 C and 20-25 g/L, respectively. Each tank is supplied with gentle blown air aeration at four points with a
one point source of pure oxygen to maintain dissolved oxygen above 100% saturation. Rotifers in each culture tank are fed
concentrated Nannochloropsis paste via peristaltic pump every 4 h. Culture Selco ® is fed to each tank at 1000 and 2200 hours.
One tank is harvested daily (three days after stocking). Rotifer density at harvest ranges from 700-1,000 rotifers/ml. Once
enumerated, rinsed rotifers are restocked into another tank containing sterilized water at a rate of 250 rotifers/ml. Enrichment
occurs in buckets containing a final volume of 15 L (salinity = 28-32 g/L). Rotifers are stocked to achieve a density of 1,500
rotifers/ml. Buckets receive blown air and pure oxygen to maintain dissolved oxygen levels above 100% saturation. The enrichment
diet (Ori-Green) is then applied and rotifers are allowed to feed for 3 h at a temperature of 26-27 C. Once enrichment
is completed, rotifers are rinsed to remove residual enrichment diet and are placed into a clean bucket containing a final volume
of 15 L. Ice bottles are then added as needed to lower the temperature to 10 C over a two hour period. Buckets are then placed
in a refrigerated cooler (with ports for blown air aeration) for cold storage up to 24 h at 5-10 C.
Artemia cysts are decapsulated on site using standard procedures and cysts are stored at 0-3 C in a hypersaline solution until
needed. Hatching occurs in two fiberglass tanks containing 175 L of sterilized water. Water temperature and salinity are
maintained at 30 C and 28-32 g/L, respectively. Each tank is supplied with vigorous blown air aeration and pure oxygen to
maintain dissolved oxygen above 100% saturation. Prior to the addition of cysts, a disinfectant (Sanocare ® ), sodium bicarbonate,
and an antifoaming agent are added. Hatching is complete in 12-14 h with typical harvest amounts of 50-90 million 1 st
instar nauplii. Artemia are then rinsed and cold stored (at a density not exceeding 2,500 Artemia/ml) as described for rotifers
or they are stocked at a rate not exceeding 300 Artemia/ml into one of two available 175 L enrichment tanks containing sterilized
water (temperature = 26 C, salinity = 28-32 g/L). Artemia are enriched for 20 h with the addition of equal amounts of DC
DHA Selco ® at T0 and T10. Harvested enriched 2 nd instar nauplii are then rinsed thoroughly and cold stored at a density not
exceeding 2,500 Artemia/ml.
13

14
RT-LAMP-IMMUNOCHROMATOGRAPHY (ICT) FOR DETECTION OF INFECTIOUS
MYONECROSIS VIRUS (IMNV) IN RESOURCE-POOR DIAGNOSTIC SETTINGS
Thales P. D. Andrade*, Kathy Tang and Donald V. Lightner
Department of Veterinary Science and Microbiology
University of Arizona
1117 E. Lowell St., Bldg.90, Rm 106
Tucson, Arizona 85721 USA
thpa@email.arizona.edu
Delays and misdiagnosis of infectious shrimp diseases are most common in marine shrimp culture zones because the equipment
necessary for pathogen detection is beyond what most diagnostic sites can afford. We report the development of an
IMNV RT-LAMP-immunochromatographic (ICT) diagnostic method for detection of Infectious myonecrosis virus (IMNV) in
resource-poor and point-of-care diagnostic settings. The RT-LAMP-ICT method combines simplified nucleic acid extraction, a
reverse-transcription isothermal amplification platform, and one-step visual colorimetric confirmation of the IMNV amplified
sequences using a generic ICT qualitative detection test strip.
IMNV-specific RT-LAMP primers were designed for detecting the structural capsid protein gene of IMNV. For purpose of comparison,
efforts were made to design RT-LAMP primers that amplify the region between 365-568 bp, which encompass the region
amplified by the real-time RT-PCR (412-545bp) (Andrade et al., 2007). The sensitivity of RT-LAMP (using two and three
primer pairs) and nested RT-LAMP (using three primer pairs) was compared by Real-time RT-PCR using TaqMan probe. The
RT-LAMP using three primers pairs was 10-fold more sensitive than when using two primers pairs (10 -4 ), and the ladders 10 -3
and 10 -2 dilutions were more intensely amplified by RT-LAMP by using three primers pairs than two primer pairs. The addition
of loop primers (use of the third primers pair) to the reaction tube increased sensitivity and specificity, and this same pattern of
results was obtained in others assays. Even though the sensitivity of the nested RT-LAMP is equivalent to that of the Real-time
RT-PCR (10 -6 ), it was not selected for adaptation to ICT because of increased time processing and aberrant amplifications.
The detection of IMNV RT-LAMP products was accomplished by using an ICT test strip. The biotin-labeled IMNV RT-LAMP
amplicons were hybridized with an FITC-labeled probe specific for the IMNV capsid gene. The hybridization product was
dotted on the spot for sample application of the ICT strip, the IMNV RT-LAMP immune complexes were captured and a red
test band was generated. Labeled immune complexes not captured on the test line overflowed the control band and were immobilized
by species-specific antibodies resulting in an intense control band.
These results clearly demonstrate that the RT-LAMP-ICT method is specific, sensitive and user- friendly. The RT-LAMP-ICT
method also can shorten the time for analysis and has potential application for IMNV diagnosis in resource-poor and point-ofcare
diagnostic settings. Additional studies are under way to determine the feasibility for this method to become commercially
available.

MULITIVARIATE ANALYSIS OF THE GROWTH PERFORMANCE OF INDIAN CARPS FED
DIETS CONTAINING DIFFERENT PROTEIN SOURCES AND LEVELS
G.V.S.R. Anjaneyulu and S.V.Sharma
Department of Statistics
Acharya Nagarjuna University
Nagarjuna Nagar – 522 510
Andhra Pradesh
India
Aquaculture is the fastest growing food production sector in recent years globally. In India Carp farming is an important
aquaculture activity and its contribution to National Inland production is about 67 percent. In recent times, statistical analysis
of production systems in India is receiving increasing attention in view of the multiple ecological and biological factors that
influence production. Fish production in ponds ranges from 2 to12MT per hectare depending on the management techniques in
practice. The present communication highlights the use of different statistical analytical techniques such as Multivariate Analysis
and Step-wise Regression in associating different parameters to productivity. This presentation is prepared in two parts. The
first case is that of fry rearing of Indian major carps under different feed regimes and their rates of growth on six types of feeds
with different protein levels. The second part is a presentation on the production ponds of different sizes, species combinations,
feeds, and water parameters. These two scenarios are analyzed using different statistical techniques to arrive at defining better
management practices for carp culture in a tropical ecosystem. Response Surface Methodology techniques are also employed in
estimating the production of fish under presently adapted farming practices by the farmers to identify what factors/ independent
variables influence the dependent variable – the Production.
1

1
REVERSE GAVAGE: A NEW STANDARDIZED TECHNIQUE FOR INFECTING SHRIMP
SUITABLE FOR CHALLENGE TESTS
Fernando Aranguren* and Donald Lightner
Department of Veterinary Science and Microbiology
University of Arizona
1117 E Lowell St.
Tucson, AZ 85721 USA
lfarangu@email.arizona.edu
Experimental infection using reverse gavage was tested in Litopenaeus vannamei through challenge studies with some of the
most current infectious diseases affecting shrimp industry (WSSV, YHV, IMNV, NHP-B and Spiroplasma penaei) were run to
determine if infection in L. vannamei could be induced through this technique.
Reverse gavage, a modified gavage or force feeding method was used is this experiment. This technique consisted of introducing
a suspension containing infectious material through the shrimp’s anus into the region of the anterior midgut and its junction
with the hepatopancreas in order to provoke an infection and reproduce River’s and Koch’s postulates in the population
tested.
Tissues infected with WSSV, YHV, IMNV, NHP-B and S. penaei were used to infect shrimp using reverse gavage. Specific
Pathogen Free SPF L. vannamei Kona line shrimp of 1.5 g were used. The inoculum was obtained by grinding the infected
tissue with saline in a ratio of 1:1 g of infected tissue: ml of saline. Then the homogenized was centrifugated to 6000 x g per
one minute. The supernatant collected in a micro tube was stained with a commercial red dye commonly used as a food color
in ratio of 3:1000, red dye:solution. Twenty uL of inoculum was introduced by reverse gavage into each shrimp by using a 20
uL automatic pipette. In the Table 1 the challenge test results for each disease are shown.
These results show that reverse gavage challenge technique can be used to induce shrimp disease with the pathogens tested. As
expected, acute mortalities were obtained with WSSV and YHV. Chronic mortalities were obtained when using NHP, S. penaei
and with IMNV. In comparison with other challenge methods, the use of reverse gavage allows infection of shrimp with a very
low amount of inoculum. Also, when compared with oral gavage, reverse gavage is less traumatic because the midgut, hindgut
and anus form a strait tube while the oral region is blocked by the hard mandibles and an esophagus that bends at a sharp angle
at its juncture with the anterior stomach. In addition, the inoculum volume can easily be adjusted using this technique.

NEW CONCEPTS FOR STUDY ON SURVIVAL, GROWTH AND REPRODUCTIVE
CHARACTERISTICS OF BRINE SHRIMP Artemia (CRUSTACEA: ANOSTRACA)
Alireza Asem
Protectors of Urmia Lake National Park Society (NGO)
Urmia, Iran
alireza_1218@yahoo.com
Many studies on survival, growth and reproductive characteristics of Artemia populations have been done with biological and
economical approach. But there is very serious problem to determine about optimum conditions for getting best biological results
according to these studies. Sometimes data and results don’t match together. For example; in some study, it was observed
that survival decrease in a definite salinity though percentage of maturity increase or offspring rise in a specific salinity but
growth reduce and etc. Therefore some new concepts and their formulae are suggested here for solve these problems. Survival
is an important item for economical and biological decisions so Coefficient of Survival is proposed as factor.
1) Coefficient of Survival: CS =
No. of remained Artemia
total No. of cultured Artemia
Now this factor can qualify growth and reproductive characteristics for any comparison.
2) Maturity Index = CS *
total No. of maturated Artemia
No. of remained Artemia
3) Growth Index = mean of total length * CS
Because all animal reproduce after maturation and for any study on reproductive
characteristics of Artemia used maturated samples so CS must calculate when all
individuals are maturated (CS%100).
4) CS%100 =
No. of maturated Artemia
Total cultured No.
5) Offspring Index = CS%100 * total offspring
6) Nauplii produced Index = CS%100 * total nauplii produced
7) Cyst produced Index = CS%100 * total cysts produced
1

1
FIRST REPORT OF MORPHOLOGICAL CHARACTERIZATION OF RARE MALES AND
SEXUAL DIMORPHISM IN PARTHENOGENETIC Artemia (CRUSTACEA: ANOSTRACA):
AN EVOLUTIONARY APPROACH
Behrooz Atashbar, Alireza Asem and Nasrullah Rastegar-Pouyani
Artemia and Aquatic Animals Institute
Urmia University, Iran
atashbarb@yahoo.com
Parthenogenetic populations of Artemia have two important characters; their reproductive mechanism is only via parthenogenesis
and also the males seldom are produced in these specimens, so, the male samples are unproductive. This is the first study on
morphological and biometrical characterization of the males and sexual dimorphism in parthenogenetic populations of Artemia
carried out in the Urmia Lake, Iran. Thirty male samples were collected during five months with less than 0.4% frequency and
were compared with thirty female samples. The morphological structure of frontal knob in the males of the parthenogenetic
population from the Uremia Lake is recognized by its subspherical shape Also the structure of furca is represented by having
two lobed with many setae (Fig 1). Twelve morphometric common characters between males and females were measured.
Principal Components Analysis shows that male and female groups are separated completely. The first and second components
show 66.45% and 14.56% of the total variation respectively; in total the two components show 81.02% of variation. Discriminant
Function Analysis confirms 100% of the original groupings. Female is bigger than the male; so, This result can prove; at
less parthenogenetic population of Artemia from Urmia Lake could keep its ancestral and evolutionary characters; because the
parthenogenetic Artemia populations are new specimens that had been separated from bisexual species of Artemia.
a) b)
Figure 1. The structure of frontal knob: subspherical shape (left) and furca: tow lobed with
many setae (right) in parthenogenetic population Artemia from the Urmia Lake.

FORMULATION AND APPLICATON OF PROBIOTIC AS BIOCONTROL AND BIOSAFETY
FOR ORGANIC AQUACULTURE SYSTEM
Dr. Ohnmar Aung* and Dr. San San Yu
Aquaculture Biotechnology Research Center
Department of Biotechnology
Mandalay Technological University
Myanmar
In this research, three species of beneficial bacteria Bacillus mesentericus, Nitrobacter winogradskyi and Streptococcus faecium
were isolated from different sources. Four species of beneficial bacteria Rhodopseudomonas palustris, Pseudomonas
fluorescens, Pseudomonas stutzeri and a yeast Saccharomyces cerevisiae were obtained form the Department of Biotechnology.
In this study, in vitro disease inhibition test of probiotic bacteria were tested with V. harveyi, A. hydrophila, V. parahaemolyticus
and E. coli. A new probiotic was formulated by using these seven beneficial microbes. They were inoculated in liquid carrier.
The initial bacterial population in liquid carrier was 8.8 ×10 9 CFU/ml. After three months, the bacterial population stored at
room temperature and refrigerator were 6.4×10 7 CFU/ml and 3.6×10 8 CFU/ml respectively. Six trials (i) 5ml/ton(probiotics-
P) (ii) 10ml/ton(P) (iii) 15ml/ton(P) (iv) 20ml/ton(P) (v) antibiotic (vi) without treatment with two replicate of each were
conducted as experiment I in 60 liter tanks and as experiment II in 1 ton tanks for larval rearing of giant freshwater prawn
(Macrobrachium rosenbergii). Survival rates, emergence time of post larvae, completion of post larvae development, and water
quality were found to be better in all trial with probiotic. Moreover, Vibrio counts in the larval rearing tanks were at minimum in
probiotic treatments. The microbial species diversity in hatchery tanks can be adjusted by adding selected beneficial bacteria to
displace deleterious bacteria. The formulated probiotic was found to have beneficial effect on water quality and disease control
as well as survival rates (%) in these experiments. Therefore the new formulated probiotic will be useful as biological control
and biosafety for organic aquaculture system.
1

20
2008 FARM BILL: IMPLICATIONS FOR THE UNITED STATES FARM-RAISED CATFISH
INDUSTRY
Jimmy L. Avery and Gary Jensen
National Warmwater Aquaculture Center
Mississippi State University
Stoneville, MS 38776 USA
javery@drec.msstate.edu
The United States Congress passed the Food, Conservation and energy Act of 2008 which was eventually enacted into law. This
legislation, commonly referred to as the 2008 Farm Bill, contains numerous provisions which are specific to aquaculture. Some
of these provisions have the potential to significantly impact the United States farm-raised catfish industry. This presentation
will highlight some of these key provisions.
Currently, the USDA Food Safety Inspection Service (FSIS) provides continuous inspection of processing of all meat, poultry
and eggs. The 2008 Farm Bill amends the Federal Meat Inspection Act authorizing FSIS to include catfish in the list of inspected
products. In addition to processing, the FSIS will take into account the conditions under which catfish are raised and
transported to the processing facility. This program is at no cost to the processor. The primary impact to the industry is that no
product FSIS classifies as “catfish” will be allowed to be imported into the United States until an equivalent inspection program
can be certified in the exporting country.
The USDA Agriculture Marketing Service (AMS) provides a grading program for meat and poultry products that has widespread
consumer acceptance. Examples include USDA Grade A eggs and USDA Prime beef. The 2008 Farm Bill establishes
within the USDA a voluntary fee based grading program for catfish. Catfish commodity organizations supported this provision
because they feel it will be an effective marketing tool for those processors that elect to participate.
The 2008 Farm Bill extends the authority of USDA Farm Services Agency to enroll commercial pond-raised aquaculture ponds
In the Conservation reserve Program (CRP). Any catfish pond which has been in operation during one of the past 5 years is eligible.
The 15-year rental contract would convey an estimated $60-$80 an acre rental rate to ponds which discontinue operation.
The program would also extend a $100 per acre, on-time, signing incentive payment and cost sharing on vegetative plantings
and field modifications. It is estimated that as much as 15,000 to 25,000 acres could be enrolled in Mississippi alone.

ALTERNATIVE USES FOR RETIRED CATFISH ACREAGE IN MISSISSIPPI
Jimmy L. Avery and Craig S. Tucker
National Warmwater Aquaculture Center
Mississippi State University
Stoneville, MS 38776 USA
javery@drec.msstate.edu
Mississippi’s commercial catfish acreage has been in decline since its peak of 113,500 acres in 2001. This decline has recently
become acute with acreage decreasing from 96,000 acres in 2006 to 80,400 acres in 2008. This unprecedented loss in acreage
is a result of low fish prices due to increased import pressure coupled with rising costs of energy and grain-based feeds. The
2008 Farm Bill tasked the USDA Farm Services Agency to increase the amount of catfish acreage eligible for enrollment in the
Conservation Reserve Program (CRP). This presentation will describe current alternative uses for commercial catfish ponds
and an estimate of potential CRP participation in Mississippi.
Currently, retired catfish acreage is either being left fallow, used to produce other agricultural commodities, or developed
into hunting operations. Fallow ponds remain flooded or are dried and disked to prevent vegetation growth. The soils in pond
bottoms contain only slightly higher levels of nitrogen, phosphorus and organic matter than the same soils (alligator/sharkey
clays) used for row-crop agriculture. Record prices for corn, soybeans, and rice have provided farmers incentive to plant these
traditional row crops in pond bottoms. A significant percentage of retired catfish ponds has been developed into personal hunting
opportunities or have been sold for the development of duck hunting operations.
Catfish ponds being converted to CRP acreage will require modification prior to vegetative planting. After draining, the existing
drainage structure will need to replaced or modified to facilitate lower water levels. Excessive vegetation will need to be
controlled by chemical application or disking. Earthen ramps for equipment access to the pond bottom will have to be constructed.
Although, it is difficult to characterize all pond situations, estimated preparation costs prior to planting could be $125
to $200 per acre.
If the proposed CRP program proves to be economically attractive, ponds that have been left fallow, have low commodity
yields, or were being used for personal hunting opportunities are likely to be enrolled. It is doubtful whether acreage that has
had significant investment in conversion to crop land or developing waterfowl habitat will be enrolled in the program. Some
catfish farmers may decide to enroll active acreage that is difficult to farm due to a remote location, poor site conditions, or high
bird predation. Of the 33,100 acres that have been retired in Mississippi since 2001, it is estimated that 13,000 to 15,000 acres
have potential to be enrollment. An additional 5,000 to 7,000 acres of active ponds may also be enrolled.
21

22
EXPERIMENTAL STUDIES ON THE PATHOGENICITY OF BACTERIAL ISOLATES FROM
TILAPIA
Sayed Awad
60 Coldeer Street Giza
Giza 12211, Egypt
sayed_awad11@hotmail.com
In this experiment an 80 clinically normal growing Tilapia (Average weight 50.0 + 10.0g.) Were divided into 8 groups each of
10 fish. The frist 3 groups were injected intra muscular each of which was injected by one isolate where each fish was received
intra muscularly 0. 2ml of 24 hrs old cytophaga broth culture containing 2x105 cytophaga clumnaris microorganism.
The fourth and fifth groups were injected intra musculary each of which was injected by one isolate where each fish was
injected intra musculary by 0.2 ml of 24 hrs old A. hydrophila broth culture containing 4x105 A. hydrophila microorganism.
The sixth group fish were injected intra musculary by 0.2 ml of 24 hrs. old p. flurscence broth culture containing 2x105
organism.
The seventh group included 10 fish was used as control group which were inoculated (I.M.) with 0.2 ml stril broth.
The last group was used also as a control group it inclouded 10 fish each of which my inoculated with 0.2ml strile cytophaga
broth intramuscularly in the back region.
Results of clinical studies:
The clinical picture of experimentally inoculated fish in the first three group nearly similar but varied only in severity.
Fading of fish colour,poor appetite sluggish rotatory movment, faliar of response to external stimuliand death. There were
oedematous swelling at the site of ocuolation which was congested and surrounded by hyperemic zone . most of fish showed
congestion and haemorrhage especially at caudal peduncl , in more developed cases the site of inoculation erupted leaving
Regarding the clinical flat ulcer
Regarding the clinical picture in fish groups 4,5,6, these could be sumerized as following .
The clinical abnormalities and death began to appear 24 hours post-inoculation, thtse included sluggish movment ,poor appetite
,impairment of equilibrium and finally loss of reflexes just prior to death.
Regarding the clinical abnormalities,there were haemorrhagic patches started as spots on the skin especially venteral abdominal
area ,caudalpeduncle and on the body fins especially at there bases. As the disease progressed. There was whitish opcity at the
free margin of the body fin especially the caudal ones which began to slough out. In addition to these symptoms, there were
also abdominal distintion of varying degree and severely inflamed, edematous.

EFFECT OF FEED RATE ON FOOD CONSUMPTION BY THE SEA URCHIN Lytechinus
variegatus
Adrian Bandolon*, Joe M. Fox, Addison L. Lawrence, John M. Lawrence and Stephen A. Watts
Texas AgriLife Research
Mariculture Laboratory
1300 Port Street
Port Aransas, Texas 78373-4200 USA
abandolon@islander.tamucc.edu
Increased demand for sea urchin roe that has led to diminished wild stocks. To alleviate fishing pressure on these species,
aquaculture production should be considered. Successful culture of sea urchins requires an in-depth understanding of their
nutritional requirements. This study was conducted to determine optimal feed rates for small (17.40g ± 1.66, n = 48) and large
(23.46g ± 1.30, n = 48) Lytechinus variegatus. A standard reference diet was offered daily at four feed rates (3.0, 2.5, 2.0, 1.5%
wet body weight as feed; bwd) to urchins over a nine-day period and unconsumed feed was collected 24 h post feeding. New
rations were estimated based upon weight determinations made every three days. Uneaten feed was quantified by weight after
drying at 90°C for 16 h. A second trial was conducted with small and large urchins to evaluate feed consumption within a 24
h period. For this trial, uneaten feed was collected at 1, 3, 6, 9, 12 and 24 h post feeding.
Results from the first trial showed that after 9 d, both small and large urchins fed at the 1.5% bwd feed rate consumed feed at
a significantly lower rate compared to those fed at other feed rates (P

24
THE EFFECT OF VITAMIN PREMIX IN EXTRUDED PLANT BASED AND FISH MEAL
BASED DIETS ON GROWTH EFFICIENCY AND HEALTH OF RAINBOW TROUT,
Oncorhynchus mykiss
Frederic T. Barrows*, T. Gibson Gaylord, Wendy M. Sealey, Lucas Porter and Charlie E. Smith
USDA - Agricultural Research Service
Hagerman Fish Culture Experiment Station
3059-F National Fish Hatchery Road
Hagerman, ID 83332 USA
Rick.Barrows@ars.usda.gov
Trout diet formulations have changed considerably in recent years and todays diets reflect changing ingredient markets, feed
processing technologies, and faster growing strains of trout. In contrast, most of the vitamin requirements for rainbow trout
were determined decades ago and whether these requirements are appropriate for modern diet formulations are unclear. For
these reasons, a study was conducted to determine the effect of vitamin levels and protein source in extruded feeds on fish
performance.
A 2 by 4 factorial treatment design was used with four vitamin premixes and two protein sources, fish meal and plant meals.
All vitamin premixes contained the same vitamins but at different levels and included; 1) 100% of NRC (1993) recommended
vitamin level (NRC), 2) NRC levels with each vitamin independently increased for potential extrusion losses (variable retention
,VR), 3) NRC levels with all vitamins increased to assume equal retention of 40% after extrusion (ER), and 4) negative control
with no vitamin premix added (None). Each of the 8 diets was fed to groups of 35 rainbow trout (4.8 g) in 150-L fiberglass
tanks (5 replicate tanks per diet, except for the negative control diets with 3 replicates) for 15 weeks. Each tank was supplied
with 6 L/min of untreated, constant temperature (14.5ºC), spring water.
Signs of a pantothenic acid deficiency were observed within 6 weeks for the fish fed the plant-based diet without vitamin
premix. Signs of a vitamin E deficiency were observed for the trout fed the fish meal-based diet without vitamin premix after
6 weeks of feeding. The two un-supplemented treatments were terminated after 9 weeks. Among the supplemented diets,
vitamin premix did significantly affect survival, feed intake, protein retetntion efficeicny (PRE), energy retention efficiency,
hematocrit and HSI, but not weight gain or FCR. Significant interactive effects between vitamin premix and protein source
were observed for survival, hematocrit, HSI, and PRE. Vitamin retentions after extrusion were similar to reported values with
the exception of vitamin A, folic acid, and thiamin. Results indicate that the vitamin levels recommended by NRC do not appear
to be adequate for young, fast growing trout fed extruded feeds. Adjustment of individual vitamins in the premix to account
for vitamin destruction during the extrusion process will maintain levels in the final feed at target levels while reducing
vitamin costs associated with over-supplementation.

UTILIZATION OF FISH PROCESSING BYPRODUCTS
P.J. Bechtel
USDA- Agricultural Research Service
245 O’Neill Building
University of Alaska
Fairbanks AK 99775 USA
bechtel@sfos.uaf.edu
The term “waste” has often been used for materials that were left over after primary food processing. If no other alternatives are
available, these materials must be disposed of in a satisfactory manner, which creates an expense for the processor. However,
a closer look finds that this “waste” can be used as the raw materials to make other valued products. Because these materials
are not now viewed as waste material they are commonly referred to as byproduct. There are many examples where yesterdays
“waste“ became a valued raw material for further processing. Enhancing the utilization of byproduct can generate additional
profit and possibly reduce environmental concerns.
The largest source of fish processing byproducts in the United States comes from the processing of wild marine fish in Alaska.
In Alaska large amounts of pollock, salmon, cod and flat fish are harvested annually resulting in over 1million metric tones of
byproducts, which can be utilized to produce a number of products including meals and oils. The processing of aquaculture
reared animals is another source of fish byproducts that can be used to make higher valued food and feed ingredients. Major
byproducts from the fish processing industry include heads, viscera, frames and skin. However, these products can be further
fractionated into organs such as gonads, livers, stomachs and other tissues, which can be used as raw materials to produce
unique and potentially valued food and feed ingredients and a host of other products.
Enhancing the utilization of fish processing byproducts can generate additional profit from these varied raw materials. There
are bio resources form wild and aquaculture harvests that can be potentially utilized for the production of interesting products.
The array or byproducts available and their potential utilization will be discussed.
2

2
HORMONE DOSE AFFECTS EGG AND LARVAL QUALITY DURING INDUCED SPAWNING
OF BLACK SEA BASS Centropristis striata WITH PELLETED LHRHa
Christopher D. Bentley*, Troy C. Rezek and Wade O. Watanabe
University of North Carolina Wilmington
Center for Marine Science
601 S. College Road
Wilmington, NC 28403-5927 USA
Bentleyc@uncw.edu
Black sea bass (BSB) Centropristis striata is an important commercial and recreational fishery species in the southeastern
US and a promising candidate for intensive aquaculture. Methods for controlled spawning of BSB are well developed, but
additional work is needed to improve the reliability of spawning and the quality of eggs produced as a basis for commercial
cultivation. The objectives of this study were to compare the effects of different doses of luteinizing hormone releasing
hormone-analog (LHRHa) on spawning performance and egg quality in BSB. Adult broodstock (N = 140) were held in twelve
1.8-m dia. controlled environment tanks in sea water (35 g/L). Broodstock were maintained at ambient photothermal conditions
until the spawning season (Apr-Jul 2008) when constant temperature (mean = 19.7) and photoperiod (13 L:11 D) were
maintained. Vitellogenic females (N = 33) were implanted with a slow release (95% cholesterol, 5% cellulose) LHRHa pellet at
mean nominal doses of 0.0 (N = 4), 5.0 (N = 10), 24.9 (N = 10), and 50.7 (N = 9) μg/kg bw. Implanted females were monitored
36 h post implant (p.i.) for ovulation using a cannula. Females containing > 50% ovulated oocytes were stripped manually
and fertilized in-vitro. Females with < 50% ovulated eggs were checked again at 40 h p.i. Spawned eggs were fertilized with
pooled milt from three running males exhibiting > 75% motility.
Female response (% of females with hydrated eggs at 36-40 h p.i.) was approximately 50% in all treatments, except for the placebo
implanted controls (0 μg/kg bw) which showed no response. Mortalities were observed in the 5 (30%), 25 (30%), and 50
(67%) ug/kg treatments. Total egg production and mean numbers of fertilized eggs produced per female were highest in the 5
μg/kg bw treatment (120.5 x10 3 , 78.7 x10 3 , respectively), intermediate in the 50 μg/kg bw treatment (96.6 x10 3 , 57.0 x10 3 ), and
lowest in the 25 μg/kg bw treatment (74.7 x10 3 , 49.8 x10 3 ). Mean numbers of yolk-sac larvae and number of larvae surviving
to 36 h post hatch per female were highest in the 5 μg/kg bw treatment (47.3 x10 3 , 31.8 x10 3 ), intermediate in the 25 μg/kg bw
treatment (11.4 x10 3 , 8.5 x10 3 ), and lowest in the 50 μg/kg bw treatment (6.9 x10 3 , 1.5 x10 3 ). Relative total egg production and
numbers of fertilized eggs produced (eggs/kg bw) were lowest in the 25 μg/kg bw treatment (77.9 x10 3 , 47.8 x10 3 ), intermediate
in the 5 μg/kg bw treatment (85.9 x10 3 , 55.7 x10 3 ), and highest in the 50 μg/kg bw treatment (107.4 x10 3 , 63.1 x10 3 ). Relative
mean number of yolk-sac larvae produced (larvae/kg bw) was highest in the 5 μg/kg bw treatment (120.5 x10 3 ), intermediate in
the 25 μg/kg bw treatment (96.6 x10 3 ), and lowest in the 50 μg/kg bw treatment (74.7 x10 3 ). Relative mean number of larvae
surviving to 36 h post hatching per female (larvae/kg bw) was significantly (P = 0.03) higher in the 5 μg/kg bw treatment (21.8
x10 3 ) than in the 50 μg/kg bw treatment (1.8 x10 3 ), while the 25 μg/kg bw treatment was intermediate (7.0 x10 3 ). The results
indicated that, within a dose range of 5 to 50 μg/kg bw pelleted LHRHa, lower hormone doses improved egg and larval quality
during induced strip spawning of BSB, with best results achieved at the lowest dose of 5 μg/kg bw.

STUDY OF LIMNOLOGICAL VARIABLES IN NURSERIES OF Colossoma macropomum
(CUVIER, 1818) AND Pseudoplatystoma corruscans (AGASSIZ, 1829)
Katia Santos Bezerra, Miguel Arcanjo Santos Neto, Mônica M. C. A Souza, Juliana F. Q. S. Santos,
João B. Pereira Neto, Alfredo Oliveira Gálvez and Luis Otavio Brito
Pernambuco Federal Rural University
Fisheries and Aquaculture Department
Dois Irmãos, 52171-900
Recife, Pernambuco, Brazil
engpescalo@hotmail.com
Ecosystem’s aquatic metabolism comprises three main levels: production, consumption and decomposition. For the aquaculture
fundament the knowledge and accompaniment of the biotic and abiotic factors for a sustainable production.
The study evaluated the dynamics of variables limnologics in ponds: Colossoma macropomun (CM) and Pseudoplatystoma
corruscans (PC), during a 24hs. The work was accomplished in the Station of Fish farming of Chesf, Paulo Afonso-BA.
The variables (O2), (T°C), pH were measured every two hours, shines and transparency. Samples of water were analysis ammonia,
nitrite, nitrate, inorganic and organic phosphate, total phosphors, chlorophyll a, identification of the plankton and phytobenthos
were collected at 09:00, 15:00 and 21:00hs.
All the variables physical-chemistries were inside of the ideal for culture species. The group phytoplankton most frequent no
CM was Cyanophyta and Bacillariophyta. while in PC was Xanthophyta.
The group phytobentic most frequent no CM was Xanthophyta, while in PC was Bacillariophyta. The group of Copepods was
the most evident zooplankton in CM and there was absence of Rotifers in PC.
Low stock density contributed to maintain the good water quality as well as the low existing renewal. In the CM treatment (tambaqui)
which receives aloctone material with great frequency, higher chlorophyll “a” concentration was detected. The presence
of copepods at this nursery can be related to the high phytoplankton’s availability in this environment.
2

2
BIOMETRIC STUDY OF MERCURY EXPOSURE ON THE EARLY LARVAL STAGES
OF BRINE SHRIMP, Artemia spp. (CRUSTACEA: ANOSTRACA)
Dola Bhattacharjee and Alireza Asem and Amin Eimanifar
Centre of Advanced Study in Marine Biology
Annamalai University
Parangipettai
India
ad_plank@hotmail.com
Artemia is being used as a rewarding tool in toxicology. Usually; the brine shrimp larvae are tested to determine the effect
of heavy metals on biological systems. The present study assessed the secondary effect of mercury at various concentrations
on the biometry of nauplii and meta-nauplii of Artemia sp. Generally, the total length and abdominal width were limited by
increasing the mercury concentrations (Fig 1. and Fig 2.). However, hormetic responses were also observed under certain
concentrations.
Fig. 1. The effect of mercury concentrations on total length
of nauplii and meta-nauplii of brine shrimp
Fig. 2. The effect of mercury concentrations on abdominal
width of nauplii and meta-nauplii of brine shrimp

GROW-OUT PERFORMANCE OF NILE TILAPIA (Oreochromis niloticus L.) FINGERLINGS
FROM DIFFERENT HATCHING SYSTEMS
Remedios B. Bolivar, Eddie Boy T. Jimenez, Roberto Miguel V. Sayco, Reginor Lyzza B. Argueza,
Hernando L. Bolivar and Russell J. Borski
Freshwater Aquaculture Center - College of Fisheries
Central Luzon State University
Science City of Muñoz
Nueva Ecija 3120, Philippines
rbolivar@mozcom.com
Tilapia hatcheries in the Philippines use various techniques in fry production that includes ponds, hapas and tanks. This study
was conducted to evaluate the growth performance of Nile tilapia (Oreochomis niloticus L.) fingerlings in earthen ponds that
were produced from three hatching systems, artificial incubation units, hapas and ponds. Treatments used in the study were
based on the source of the fry. Treatments were the following: I –artificial incubation-hatched fry, II – hapa-hatched fry, III
– pond-hatched fry and IV – combination of hatched fry by stocking 33.3% from each hatching source. Each treatment was
replicated three times. Twelve (12) 500 m 2 ponds were used for rearing and each was stocked at 4 fish m -2 or 2000 fingerlings
pond -1 . Fingerlings for treatments I, II, III and IV had an average body weight of 0.292 g, 0.308 g, 0.320 g and 0.307 g,
respectively. The average body length measured were 2.56 cm, 2.63 cm, 2.66 cm and 2.50 cm for Treatments I, II, III and IV,
respectively. Analysis of variance showed no significant difference among groups with respect to initial size.
After four months of rearing in ponds, final weight and length did not differ among treatments (P>0.05). Total harvested fish
per replicate per treatment were counted for determination of percentage survival. Treatment IV had the highest survival with
59.30% followed by Treatment II with 58.63%, Treatment I with 56.63% and Treatment III with 56.30% survival. There
were no significant differences in survival among groups. Artificial incubation-hatched fry had the highest extrapolated yield
(kg/ha) with 3890.67 kg/ha followed by hapa-hatched fry with 3797.49 kg/ha, combination of hatched fry with 3488.96 kg/ha
and pond-hatched fry with 2995.15 kg/ha. However, these differences were not statistically insignificant. It is concluded that
fingerlings reared from different hatching systems (artificial incubation units, hapas and ponds) do not significantly affect final
length and weight, survival and yield of Nile tilapia during pond growout.
2

30
STRESS EFFECTS IN CHANNEL CATFISH (Ictalurus punctatus) FRY ON POND SURVIVAL
Natha J. Booth and Brian C. Peterson
Catfish Genetics Research Unit, ARS-USDA
141 Experiment Station Rd.
P.O. Box 38
Stoneville, MS 38776 USA
natha.booth@ars.usda.gov
Results from previous studies suggested that channel catfish (Ictalurus punctatus) fry that were housed in very high traffic areas
from hatch until reaching 9 g in size were able to survive experimental challenge with Edwardsiella ictaluri better than siblings
who were hatched and raised in very low traffic areas. In order to determine if physical stress in channel catfish fry could affect
survival in ponds, three week old channel catfish fry from three different families were divided into two groups. One group
was physically stressed daily for 14 days, while the other was left undisturbed except for routine feeding. Whole body cortisol
measurements were taken at Day 0, Day 7, and Day 14 in both the stressed fish and the un-treated controls. On Day 15, the
fish from individual tanks were transferred to individual cages (1.2 m by 20.3 cm cylinders) in three separate ponds where they
remained for 21 days. Weights of fish from each treatment were taken at the time of transfer to cages and again at the end of
21 days. At the time of transfer to cages, the unstressed control fish were significantly higher in weight than the stressed fish (P
< 0.05). There were no significant differences in final weight of the fish. Overall weight gain at the end of the study, however,
was significantly higher in the stressed fish (P < 0.05). Overall percent survival for the two groups of fish was not significantly
different: 97.1% and 96.7% survival for the stressed and non-stressed groups, respectively. Channel catfish fry must be able
to tolerate physical stress resulting from handling during transfer from hatcheries to the ponds. This study demonstrates that
channel catfish fry are able to tolerate long term physical stress with no deleterious effects, and once the stress is removed are
able to grow at rates greater than that of their unstressed siblings.

INNOVATIVE USES FOR UNDER-UTILIZED SALMON TISSUES IN ALASKA
Cindy Bower* and Katie Hietala
USDA Agricultural Research Service
PO Box 757200
Fairbanks, AK 99775-7200 USA
Cindy.Bower@ars.usda.gov
The increased utilization of high quality fish by-products is receiving more attention as processors look for ways to expand
their markets. Fish meals and specialized fractions that can act as palatability enhancers and attractants for aquaculture feeds
are an important use, but fish by-products might also attract interest as functional foods affecting vitality and health in humans.
Potential exists for innovative products enriched with high-value fish proteins and “omega-3s” obtained from non-fillet components
of fish. However, to ensure success, underutilized fish by-products must be safely preserved. In this study, heads from
pink salmon (Oncorhynchus gorbuscha) were smoke-processed and/or fermented using food-grade lactic acid bacteria (LAB),
and the resultant oils and tissues were characterized.
Preservation of ground salmon heads through pH reduction using LAB was found to confer antimicrobial activity against
Gram-negative bacteria. Control samples did not receive LAB inoculations, relying entirely on endogenous proteolytic enzymes
to break down the tissues without acidification. Bacterial cell counts and lactic acid concentrations were recorded as a
measure of LAB viability. All samples were analyzed for moisture, ash, and lipid composition. Control samples were unable to
prevent spoilage of salmon by-products, whereas fermented salmon tissues stabilized at pH 4.5 for 120 days.
Oils extracted from smoked salmon heads retained valuable long-chain polyunsaturated fatty acids (PUFAs) in quantities equal
to frozen controls despite the high temperatures (95 °C) and extended heating times (5 hours) associated with smoking fish.
Additionally, these smoke-processed oils contained fewer products of oxidation than their unprocessed counterparts, suggesting
that natural antioxidants are imparted during the smoking process.
Salmon heads were also preservedusing a combination of smoke-processing and acidification with LAB. Wood smoke contains
com-pounds that lower the pH of salmon tissues. A further decrease resulted from LAB addition.
Increased utilization of processing discards promises environmental and economic benefits while con-serving valuable fish
resources.
31

32
USE OF AQUAFLOR TO CONTROL MORTALITY DUE TO COMMON FISH DISEASES IN
FRESHWATER-REARED SALMONIDS
James D. Bowker
U. S. Fish and Wildlife Service
Aquatic Animal Drug Approval Partnership Program
4050 Bridger Canyon Road
Bozeman, MT 59715 USA
jim_bowker@fws.gov
Aquaflor ® (50% active florfenicol; Type A medicated feed, sponsor – Schering Plough Animal Health Corporation, Summit NJ)
has recently been approved for use in U. S. aquaculture for the following claims: Administer in feed at a dose of 10 mg florfenicol/kg
fish body weight for 10 consecutive days to control mortality due to (1) enteric septicemia of catfish (causative agent,
Edwardsiella ictalurid) in catfish, and (2) coldwater disease (causative agent, Flavobacterium psychrophilum) and furunculosis
(causative agent, Aeromonas salmonicida) in freshwater-reared salmonids. It’s anticipated that Aquaflor ® will be approved for
additional claims in the U. S., including use to control mortality due to F. columnare in freshwater-reared salmonids. Approvals
were the result of contributions from several public data-generating partners, including the USFWS’s Aquatic Animal Drug
Approval Partnership (AADAP) Program. The AADAP Program and their partners conducted seven controlled field effectiveness
studies in support of current and future approvals for use of Aquaflor ® to control mortality due to common fish diseases
in freshwater-reared salmonids.The field efficacy studies were conducted at hatcheries that experience recurring, somewhat
predictable outbreaks of the diseases of concern (e.g., coldwater disease, furunculosis, columnaris disease). Studies comprised
a short acclimation period, a 10-d treatment period, and a 14-d posttreatment period. Results from each study showed that mean
cumulative mortality at the end of the posttreatment period was significantly lower (P < 0.05) in treated tanks than in control
tanks and that the concentration of florfenicol in feed was within ±20% of the target concentration.
Table 1. Mean percent cumulative
mortality (MCM) in treated and control
tanks at the end of each field
effectiveness study. Note, Fur =
furunculsis; CWD = coldwater disease;
col = columnaris.
Fish
species Disease
MCM
treated
(%)
MCM
control
(%)
Coho Fur 11.1 30.3
Chinook Fur 16.2 94.4
Coho Fur 17.4 30.0
Steelhead CWD 2.0 5.0
Cutthroat CWD 75.0 94.0
Coho Col 47.3 66.7
Rainbows Col 18.4 30.4
a
P-value = 0.0551
a

STATISTICAL DESIGN AND ANALYSIS OF STUDIES OF EFFECTS OF FERTILIZERS AND
OTHER AMENDMENTS ON PRODUCTION AND WATER QUALITY IN PONDS
Claude E. Boyd
Department of Fisheries and Allied Aquacultures
Auburn University, AL 36849 USA
boydce1@auburn.edu
Research on fertilizers and other pond water quality amendments usually seeks the optimum application rate for these materials.
Traditionally, aquacultural investigators have borrowed the agronomy field plot technique of establishing two or more
application rates and a control and replicating each several times. The results usually are tested for significance by analysis of
variance and range tests. In agronomy, it is possible to establish any number of field plots in areas with different types of soil
at a minimum cost. Ponds for aquaculture research are comparatively expensive. There are relatively few aquaculture research
stations, and at most, competition by different projects for ponds is great. It seldom is possible to design an experiment with
many treatments and replications. Moreover, ponds on a given station tend to be constructed on similar soil and filled from a
common water supply. In spite of this, variation in production and water quality among fertilized ponds at the same location
is high. Coefficients of variation for fish production in fertilizer experiments typically are 25 to 30% as compared to 5 to 10%
in agronomic fertilizer trials.
Recently, an alternative of establishing a large number (6 to 12) of un-replicated or duplicate treatments in ponds and treating
the results by regression analysis was evaluated. In a single experiment conducted during one growing season, the optimum
phosphorus fertilization rate in sunfish ponds at the E. W. Shell Fisheries Center at Auburn University was determined to be
3 kg/ha per application. This is essentially the same result that had taken several years to establish in typical, replicated experiments.
Similar findings resulted when this alternative approach to design and statistical analysis was applied to nitrogen
and potassium fertilization trials. It also was found that the procedure was equally useful in evaluating the influence of pond
amendments on water quality.
33

34
ENERGY USE FOR MECHANICAL AERATION OF AQUACULTURE PONDS
Claude E. Boyd
Department of Fisheries and Allied Aquacultures
Auburn University
Auburn, AL 36849 USA
boydce1@auburn.edu
Pond culture of channel catfish and marine shrimp has followed similar patterns. Production was increased through use of
feeds, and resulting problems with low dissolved oxygen (DO) concentration were initially solved through water exchange
(shrimp) and application of emergency aeration with tractor-powered aerators (channel catfish). Floating, electric aerators were
found to allow better control over DO concentration and permit greater production. Mechanical aeration use has increased
steadily, and at some intensive farms, pond aeration rates are as high as 6 to 8 hp/ha for channel catfish and 18 to 24 hp/ha for
marine shrimp. Mechanical aerators for channel catfish farming are based on a highly-efficient design developed at Auburn
University, but shrimp farmers mostly use less efficient aerators. Moreover, aerators tend to be operated nearly continuously in
shrimp ponds, but mainly at night in channel catfish ponds. Typically, aerators are operated about 1,000 hr per crop in catfish
ponds and 2,000 hr per crop in shrimp ponds, and net production is around 8,000 kg/ha for channel catfish and 14,000 kg/ha for
shrimp. Energy input is approximately 4,500-6,000 kW•hr/ha (0.56 to 0.75 kW•hr/kg) for channel catfish and 27,000 to 36,000
kW•hr/ha (1.93 to 2.57 kW•hr/kg) for marine shrimp. Energy use for aeration in shrimp culture could be reduced by 25 to 50%
by use of more efficient aerators. It also might be possible to lessen energy use in shrimp culture by reducing aerator operation
when DO concentrations are near or above saturation between mid morning and mid afternoon. In both types of aquaculture,
and especially in shrimp culture, some farms are investigating the use of heterotrophic systems in which bacterial floc largely
replaces phytoplankton. Around 30 to 40 hp/ha of aeration are used in “floc” systems for shrimp, and production of 20,000 to
24,000 kg/ha or more per crop is possible. The energy requirement per unit of shrimp production using Asian-type paddlewheel
aerators in “floc” systems is about the same as in “green-water” systems.
In both “green water” and “floc” systems, wastes are oxidized in ponds, and this consumes a portion of the DO supplied by
aerators. A possible innovation would be to remove wastes from culture systems to lessen DO requirements, and oxidize waste
externally using less energy-intensive technology than mechanical aeration.

SEEPAGE LOSSES FROM EARTHEN AQUACULTURE PONDS
Claude E. Boyd
Department of Fisheries and Allied Aquacultures
Auburn University, AL 36849 USA
boydce1@auburn.edu
Earthen aquaculture ponds seep through their bottoms, beneath embankments, and around water control structures. Several
methods have been used to measure seepage as follows: water balance (water level change measured with a staff gauge – evaporation
during periods without inflow); seepage meters; water level change in vertical pipes extended into the pond bottom and
capped; aerodynamic technique (extrapolation of the regression line of daily water level decline (Y) and the product of average
daily wind velocity and vapor pressure deficit (X) to the Y-intercept or zero evaporation). The seepage meter and capped pipe
methods give highly variable results from one place to another in pond bottoms, and they can only be used to measure seepage
through pond bottoms. The aerodynamic technique is tedious. The water balance method seems more applicable than other
procedures for use in aquaculture, but data are needed from a class A evaporation pan located in the vicinity. It also usually
requires 3 to 5 days to obtain a reliable estimate of water level decline with a staff gauge. An modification of the water balance
method was developed that relies on a small, floating, evaporation pan and measurement with a hook gauge of pond water level
decline and evaporation from the floating pan. This modification can be used to obtain a seepage estimate after 24 hr.
A literature review of seepage estimates from 39 ponds revealed a range of 1.5 to 176.5 cm/month. Average seepage was 23.1
cm/month with a standard deviation of 33.7 cm/month. The data were categorized as follows: normal seepage, 40 cm/month. Methods for reducing seepage will be summarized.
3

3
AMMONIA EXCRETION AND OXYGEN CONSUMPTION RATES OF JUVENILE
POMPANO ON SIX EXPERIMENTAL POULTRY BY-PRODUCT DIETS
Seth Bradley, Tim Pfeiffer, Ron Malone and Marty Riche
Louisiana State University
Department of Civil and Environmental Engineering
Baton Rouge, LA 70808 USA
sbradl3@lsu.edu
Florida pompano, Trachinotus carolinus, is a high market value marine finfish. Recirculating aquaculture systems present a
sustainable method for growing high densities of pompano inland in a low-salinity environment. This study determined the
24-hr ammonia excretion and oxygen consumption rates for juvenile pompano feed a pelleted feed, of which five contained
poultry by-product meal (PBM) which is a cheaper and more readily available protein by-product than fish meal. Eighteen
experimental systems with temperature control, biofiltration, UV sterilization, and supplemental aeration and oxygenation for
137 liter tanks were each initially stocked with 75 juvenile pompano weighing approximately 7-9 g. each. The pompano were
maintained on a 2.0 mm, 50-15 slow sinking finfish starter diet (Ziegler Brothers, Hardners, PA) at approximately 18-25 g daily.
Three randomly chosen units were fed one experimental diet for 5 days before the fish in the units were evaluated over a 24 hr
period for ammonia excretion and oxygen consumption. Every four hours the recirculating water flow and aeration / oxygen
supplementation in the three systems were shut down. Dissolved oxygen and ammonia concentration were measure at the start
of the shut down period and thirty minutes later. At the end of the 24 hr period the pompano in each of the systems were counted
and weighed. Oxygen consumption and ammonia excretion rates were determined in terms of g O 2 consumed per hr-g fish and
g TAN excreted per hr-g fish for each four hour time period. Results of O 2 consumption and ammonia excretion rates of the
experimental and control diets for the juvenile pompano will be presented.

EVALUATION OF BODY AND FILLET COMPOSITION, AND HEMATO-IMMUNE
PARAMETERS FOR SUNSHINE BASS FED PRACTICAL DIETS CONTAINING SOYBEAN
MEAL AND POULTRY BY-PRODUCT MEAL AS TOTAL REPLACEMENTS OF FISH MEAL
AT VARIOUS (32, 36, AND 40%) PROTEIN LEVELS
Yolanda J. Brady *, Linda S. Metts, Kenneth R. Thompson and Carl D. Webster
Fisheries & Allied Aquacultures
Auburn University
Auburn, Al 36849 USA
bradyyj@auburn.edu
A feeding trial was conducted in 12, 0.04-ha earthen ponds, with juvenile (mean individual weight 35 g) sunshine bass (Morone
chrysops X M. saxatilis) to evaluate body composition, fillet composition, and immune function responses when fed practical
diets containing soybean meal (SBM) and poultry by-product meal (PBM) as complete replacements for fish meal (FM) and
simultaneously reducing dietary protein (32%, 36%, and 40%) compared with fish fed a commercial high-quality finfish diet
containing 30% FM, and 40% crude protein (Diet 4).
After 459 days, the intraperitoneal fat (IPF) and percent hepatosomatic index (HSI) in sunshine bass was not significantly
different (P > 0.05) among treatments, averaging 5.9% and 2.53, respectively. However, fillet yield (PFW) of fish fed diet 2
(30.3%) and diet 3 (30.3%) was significantly (P < 0.05) higher than of fish fed diets 1 (28.3%) and 4 (29.2%). There were no
significant differences in moisture, protein, lipid, and ash (as-is-basis) of fillet muscle among treatments and averaged 74.3%,
21.9%, 4.7%, and 1.2%, respectively. Likewise, immune function responses were measured and no significant differences
were found in total serum protein, total immunoglobulin, lysozyme activity, natural hemolytic complement activity, and percent
hematocrit among treatments and averaged 57.8 mg/mL, 5.1 mg/mL, 93.4 ug/mL, 96.0 Unit/mL, and 60.6%, respectively
(Table 1).
Results indicate that there were little differences in fillet yield percentage, and no differences in IPF percentage, percent HSI,
proximate fillet muscle, and immune function responses of sunshine bass fed a diet containing 0% menhaden FM, 25% SBM,
24% PBM, and 32% CP compared to fish fed a high-quality commercial finfish diet. This is the first study to report that complete
substitution of menhaden FM is possible when sunshine bass are grown in ponds to market- size if alternative protein
sources, i.e., SBM and PBM are provided in the diet, and dietary CP levels can be reduced to 32%. Results from this study may
help reduce diet costs for sunshine bass producers and feed mills, thereby increasing profitability.
37

3
OPTIMIZING SURVIVAL AND GROWTH OF FIRST-FEEDING COLORADO RIVER
CUTTHROAT TROUT – DIET, TEMPERATURE, AND DENSITY IMPLICATIONS
Mandi M. Brandt*, Christopher A. Myrick and F. T. Barrows
Department of Fish, Wildlife, and Conservation Biology
Colorado State University
Fort Collins, CO 80523-1474 USA
Mandi.Brandt@colostate.edu
Cutthroat trout formerly occupied an expansive area of western North America. However, due to habitat loss, over-fishing, and
introduction of non-native trout they have experienced population declines leading to threatened/endangered status or extinction.
Propagation programs are now necessary for recovery or restoration of these subspecies. Optimal culture conditions for
cutthroat trout are largely unknown, so these fish are often reared using rainbow trout diets and culture techniques leading to
poor hatchery performance relative to these other salmonids. This study is designed to identify diets, rearing temperatures, and
rearing densities that produce optimal growth, condition, and survival of Colorado River cutthroat trout (CRCT; Oncorhynchus
clarkii pleuriticus), which may be applicable to other inland cutthroat trout subspecies.
We separately determined the effects of diet and rearing temperature on CRCT performance. Colorado River cutthroat trout
were reared in 10ºC water and fed one of five commercial diets, two experimental diets, or a commercial diet combined with
Artemia in a 119-day blinded feeding trial. Fish fed the Artemia-supplemented diet were 17% heavier, and had 6% higher
survival than fish fed non-supplemented diets. There were also significant differences amongst non-supplemented diets, with
soft-moist diets yielding lower performance and high protein, low moisture diets producing larger fish. Skretting Bio-Vita Fry,
a premium salmon fry diet produced the highest growth. There were significant differences in survival (60-86%) amongst the
diets.
A 120-day temperature study used the top two performing diets
(Skretting Bio-Vita Fry and Rangen Regular Trout, both supplemented
with Artemia) from the diet experiment in conjunction
with temperatures of 12.5-20ºC. The optimal diet-temperature
combination (Skretting Bio-Vita Fry – 16.4°C) for growth was
determined using quadratic regression (Figure 1). Survival was similar
between diets but decreased with increasing temperature.
The optimal diet-temperature combination is being used in a 120day
rearing density experiment (150, 300, 450, and 600 fish/tank).
Day 60 results show no statistically significant differences in growth,
condition, or survival among treatments.

EFFECTS OF PHEROMONAL STEROIDS OR PROSTAGLANDIN GIVEN TO FEMALE
CHANNEL CATFISH Ictalurus punctatus ON ATTRACTING MALE BLUE CATFISH Ictalurus
furcatus OR MALE CHANNEL CATFISH
Jason S. Broach*, Shelley E. England and Ronald P. Phelps
Department of Fisheries and Allied Aquaculture
Auburn University
Auburn, AL 36849 USA
jzb0007@auburn.edu
Hybrid catfish, female channel catfish (Ictalurus punctatus) X male blue catfish (Ictalurus furcatus), exhibit better traits for
commercial aquaculture such as faster growth, better feed conversion, increased disease tolerance, etc, than do channel catfish.
However, the shortage and inconsistency of seed production of these hybrids has hindered its adoption by the industry. Traditional
methods of spawning catfish such as open pond and pen spawning has had limited success with hybrid spawning due to
some type of behavioral or environmental inhibiting factor. One of the primary reasons thought to cause this low success is a
lack of the male blue’s interest and attraction to the female channel. This study was conducted to determine if a blue and channel
male’s attraction to channel females could be increased via pheromonal steroid injections of either 17α, 20β-dihydroxy-4-pregenene-3-one
(17,20β-P) or its glucorinated form (17,20β-P-Glucosiduronate), or prostaglandin F-2α (PGF-2α).
Two trials were conducted using blue and channel males with average weights of 8.2 ± 0.91 kg and 3.9 ± 0.92 kg respectfully,
stocked into two separate 644 m² ponds at 20 males per pond. In each trial, four sets of six channel females with an average
weight of 4.63 ± 1.0 kg were injected intraperitoneally with an ethanol solution containing either 0.0 mg/kg or 0.5 mg/kg of
one of the following: 17,20β-P; 17,20β-P-Glucosiduronate; or PGF-2α. Females were held separately in large traps constructed
from 121 L plastic barrels spaced 6.4 ± 0.65 m apart in the ponds for a total of 12 females per pond. Traps were checked at 6,
12, 24, 48, 72, and 96 hours post-injection, and the number of males caught per female recorded. Logistic regression analysis
was used to generate maximum likelihood estimates of a male being captured based upon male species, female strain, hormone
injected, and post-injection time.
Channel males were attracted to PGF-2α injected females on 22.9% of their given opportunities, and were attracted to control
females only 1.4% of the time. Blue males responded to 17,20β-P-Glucosiduronate and PGF-2α injected females 8.3% of their
given opportunities for each treatment, but were not attracted to the control females. Males were 9.8 times more likely to be
captured at 48 hours post-injection than at either 12 or 24 hours post-injection. There was significant variation in response
among individual fish. One PGF-2α injected female attracted 66.7% of the channel males trapped in that trial. Only 15% of blue
males used were attracted to channel females, but those blue males were attracted on multiple occasions. These results suggest
that PGF-2α injections into channel females may enhance a blue and channel male’s attraction to channel females.
3

40
LARVAL DEVELOPMENT AND YOLK UTILIZATION OF THE COCAHOE MINNOW
Fundulus grandis IN RESPONCE TO VARIATIONS OF TEMPERATURE AND SALINITY
Charles A. Brown*, Craig Gothreaux and Christopher C. Green
Aquacuture Research Station
Louisiana State University
Baton Rouge, LA 70820 USA
CABrown@agcenter.lsu.edu
The marine baitfish industry holds incredible economic potential for the coastal regions in the U.S., providing an opportunity
for millions of dollars in revenue. Currently most marine baitfish are harvested from the wild, with a relatively low number
of aquaculture facilities providing bait to marine and estuary anglers. Research centered on marine baitfish is developing, with
many aspects of farming still needing to be refined for optimal and efficient conditions. Cocahoe minnows (Fundulus grandis)
are a preferred baitfish when fishing for popular sportfish; however, the majority of supplies that come from wild harvests are
inconsistent and unable to meet the demands that arise in late summer and fall.
This study serves to determine the influence of incubation temperature and salinity on egg-yolk utilization and size at hatch
within the shortest incubation time. Variables such as body length, body depth and yolk area at hatch have been viewed as critical
aspects for larval survival at first feeding and beyond. Incubation temperatures were maintained at 20, 22, 24, 26, and 28° C
using incubation chambers. Incubation trays filled with 3-cm of water at salinity concentrations maintained at 5g/L and 10g/L.
Newly fertilized eggs were placed in the incubation trays and checked daily for the presence of newly hatched larva. The larva
were collected and placed in 10% formalin solution and dated. Each individual was placed on their sagittal axis in a propylene
glycol solution and photographed with a digital camera mounted on a dissecting microscope interfaced with a computer. Each
image was analyzed by image analysis software to determine the length, body depth and yolk area of each larva.
We anticipate that higher temperatures will produce larva with the lowest incubation time and greatest yolk volume. However,
if the larva are not developed enough, i.e. lacking a fully formed mouth or underdeveloped tail musculature to support the body
during swimming, a faster hatch time may not be beneficial to the larva. These data will support later investigations of development
in this species to better facilitate larval rearing strategies.

EFFECTS OF DIETARY PREBIOTICS ON MICROBIAL COMPOSITION AND INTESTINAL
STRUCTURE OF JUVENILE RED DRUM Sciaenops ocellatus FED SOYBEAN-BASED DIETS
J. Alejandro Buentello*, Camilo Pohlenz, Gary Burr and Delbert Gatlin, III
Department of Wildlife and Fisheries Sciences
Texas A&M University System
College Station, Texas 77843-2258 USA
abuentello@tamu.edu
Prebiotics are non-digestible functional ingredients that beneficially affect the host by stimulating the growth and/or activity
of one or a limited number of bacteria in the gastrointestinal (GI) tract, thus improving the host’s enteric health. However,
prebiotic use also has proven to benefit the rapidly changing intestinal morphology of juvenile vertebrates by preventing villous
atrophy and eliciting cyto-protective effects on sloughing cells of the GI mucosa. Prior experiments in our laboratory
have demonstrated desirable effects of dietary prebiotic supplementation on red drum including enhanced nutrient availability,
improved growth performance, feed efficiency and disease resistance. Therefore, this experiment was conducted to assess
the effects of fructooligosaccharide (FOS) in the form of inulin, mannanoligosaccharide (MOS), transgalactooligosaccharide
(TOS), and GroBiotic ® -A (GBA) on gut structure and intestinal changes of microbial populations of juvenile red drum fed
soybean-based diets.
Experimental fish (10.9 ± 0.2 g/fish, initial weight) were acclimated for 2 weeks to the trial conditions and fed a basal diet in
which half of the dietary protein (40%) was provided by soybean meal with the other half from menhaden fishmeal. Experimental
diets incorporated FOS, MOS, TOS or GBA at 1% by weight to the basal diet. These diets were formulated to satisfy
all known nutrient requirements of red drum, provided equal digestible energy, and were fed to apparent satiation, twice daily
to triplicate groups of fish. Brackish water (10 ppt) was recirculated through 110-L aquaria at 1 L/min and kept at 26 ± 1 C,
with approximately 100% oxygen saturation and < 0.5 mg/L unionized ammonia. After a 4-week period, proximal and distal
intestine samples from three fish per aquarium were collected for determination of selected histometric indicators. Each sample
was externally and internally washed with NaCl 0.9% to remove the intestinal contents. Samples were individually transferred
to jars containing 10% buffered formalin, for fixation. After a 15-day fixation period, samples were embedded in paraffin,
sectioned to a 2 to 5-μm thickness, mounted on glass slides, and stained with hematoxylin and eosin. Denaturing gradient gel
electrophoresis (DGGE) analysis including DNA sequencing and real-time polymerase chain reaction analyses were also utilized
to characterize changes in GI tract microbiota upon prebiotic treatment. Data on microvilli height, width and density of
proximal and distal intestine are under analytical review. Based on preliminary assessment of these data, it appears that GBA,
MOS and TOS significantly altered the microbial population structure in the GI tract of juvenile red drum. Also, prebiotic
inclusion appeared to increase microvillous height and density.
Taken together, these results suggest that inclusion of these prebiotics at 1% of diet is adequate to elicit structural changes while
stimulating villi growth in the fish intestine. Likewise, prebiotic treatments appear to selectively stimulate the proliferation
of certain bacterial species. This information is likely to facilitate development of alternative strategies, including prebiotics,
probiotics and synbiotics, to improve fish growth and health management.
41

42
FISH MEAL REPLACEMENT BY PLANT PROTEIN INGREDIENTS IN SALMONID FEEDS:
A META-ANALYSIS OF PUBLISHED STUDIES TAKING INTO ACCOUNT NUTRITIONAL
ADEQUACY, GROWTH PERFORMANCE, AND NUTRIENT UTILIZATION
K. Hua and D.P. Bureau*
UG/OMNR Fish Nutrition Research Laboratory
Dept. of Animal and Poultry Science
University of Guelph
Guelph, ON, N1G 2W1, Canada
dbureau@uoguelph.ca
Economical and environmental constraints are inciting feed manufacturers to replace fish meal by plant protein ingredients in
salmonid fish feeds. Numerous published studies have shown that a large proportion of the fish meal can be replaced by plant
protein sources without deleterious effects on growth performance, nutrient utilization, and health of salmonid fish. However,
other studies have resulted in significantly lower growth performance of the fish, despite the fact that the experimental diets
appeared to be nutritionally adequate. The goal of this project is to re-evaluate the results of published studies on fish meal
replacement by plant protein sources through a systematic and statistically sound meta-analysis approach.
An extensive survey of literature on the use of plant protein ingredients in fish feeds was conducted. A sub-set of studies conducted
with rainbow trout and deemed to be scientifically sound were selected. Information on diet composition, growth performance
and nutrient deposition of the fish was compiled into a database (referred to as “Modeling Dataset”). A “Feed Evaluation
Model” was constructed by incorporating components of an “Ingredient Composition Database” into a diet formulation
spreadsheet. Up-to-date, “state-of-the-art”, information on essential amino acids, essential fatty acids, and minerals requirements
of rainbow trout was included in the Feed Evaluation Model. Digestibility sub-models for starch, lipids and phosphorus
were developed and incorporated into the Feed Evaluation Model to account for the chemical state and levels of these nutrients,
interactions between nutrients, and the effect of environment factors (e.g. water temperature).
The digestible nutrient contents of the experimental diets, as estimated by the Feed Evaluation Model, will be assessed for
nutritional adequacy against the up-to-date information on nutrient requirements of rainbow trout. The relationship between
growth and/or nutrient (protein, lipid and phosphorus) deposition of the fish and various factors (e.g. sources and levels of plant
proteins, predicted nutritional adequacy or degree of deficiency, initial body weight of the fish, etc.) will be examined statistically.
This approach will help determine if nutritional deficiencies or other unforeseen or overlooked factors (e.g. putative
nutrient deficiencies or effect of anti-nutritional factors, poor growth, etc.) could explain the results of the various studies. The
meta-analysis will help establish reliable nutritional and feed formulation guidelines when using high levels of plant ingredients
in salmonid fish feeds.

SOLDIER FLY PREPUPAE AS AN ALTERNATIVE FEED FOR CHANNEL CATFISH
G. J. Burtle*, D. C. Sheppard and G. L. Newton.
Animal & Dairy Science
University of Georgia
Tifton, GA 31793 USA
gburtle@uga.edu
Black soldier fly, Hermitia illucens, produce a harvestable prepupae that has potential as a substitute for fish meal or other
animal protein sources in diets for fish, such as channel catfish. Dried black soldier fly prepupae contains 42% CP (Met 0.83%,
Lys 2.21%, Cys 0.31%) and 35% lipid (lauric 53.4%, linoleic 8.8%). Calcium and phosphorus are 5.0 and 0.88% respectively
when grown on swine manure. Lipid and mineral composition change considerably as the soldier fly larval diet is changed.
For example, omega three fatty acids can be increased by adding materials that contain these lipids to the larval diet. Soldier fly
larvae can utilize plant products to provide a high quality ingredient for carnivorous fish and crustaceans as outlined in Goal 5
of the PPA Strategic Plan. Channel catfish (10 g) were fed diets containing 0, 7.5, 15, 22.5 or 30% soldier fly Prepupae Meal
(SF) with the 0% SF diet containing 8% menhaden fish meal and soybean meal, wheat middlings, corn, and soy oil varied to
make the diets similar in energy (DE, 2,934+ 75 kcal/kg) and protein (32.09+ .09% CP). Growth after eight weeks was similar
(P

44
FULL FAT SOYBEAN MEAL FOR BAITFISH DIETS
G. J. Burtle
Animal & Dairy Science
University of Georgia
Tifton, Georgia 31793 USA
gburtle@uga.edu
A diet made entirely of full fat soybean meal is proposed for baitfish, especially golden shiners Notemegonus chrysoleucas, due
to the need for lipid that has been shown previously for baitfish. Full fat roasted soybean meal (FFS) contains approximately
18% lipid and may be of benefit to golden shiners that have been reported to benefit from diets containing at least 13% lipid.
Soybeans are roasted in order to reduce the effect of anti-nutritional components. The meal is ground to a particle size that is
small enough for the baitfish to consume and slowly sinks when applied to water. Diets in the meal form, that contain water
soluble nutrients, may leach nutrients when applied to water and before fish can consume diet particles. This study compared
FFS to a control diet containing a variety of ingredients and supplemental vitamins and minerals. Aquaria in a recirculating
water system were stocked with 10 golden shiners per tank in triplicate tanks. Golden shiners in each tank were fed to satiation
by observing left over feed in the bottom of each tank. After 12 weeks, growth was similar (P

ECONOMICS OF A SOFT-SHELL CRAYFISH OPERATION
Nathan Bussen* and Siddhartha Dasgupta
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
Nate.bussen@kysu.edu
Soft-shell crayfish production is a small-scale enterprise practiced in states such as Louisiana, Indiana, Kentucky, etc. Softshell
crayfish are crayfish such as Procambarus clarkii, or Orconectes rusticus, or Orconectes immunus that have been induced
to molt. Farmers obtain adult crayfish from aquaculture sources or commercial fishermen. These starter crayfish are fed in
tanks and induced to molt by controlling feed, water quality and photoperiod. This is followed by chilling the soft-shell crayfish
into a hibernating state in preparation for sales activities.
In Kentucky and Indiana soft-shell crayfish production usually occurs in a flow-through tank system and facilities usually include
a climate-controlled building and water supply sources via ponds and wells. Production occurs in summer during which
much labor is required to monitor crayfish for signs of molting. Each batch of crayfish is kept from 14-21 days for molting to
occur. Interviews with farmers have indicated that soft-shell crayfish, frozen and/or live, are sold to restaurants as food and as
bait for sport fishing.
Initial capital investment of $175,000 is necessary for a relatively small-scale operation. Annual operating cost was approximately
$129,896, of which 8% is for starter crayfish, 2% for feed, and 38% for hired labor. At a production level of 400 softshell
crayfish/day (per batch survival rate = 50%) a profit of $71,248 per year can be expected. However, production is dependent
on how well the farmer manages the organisms: for example, a 20% survival rate will result in a loss of $52,520/year; a
40% survival rate would cause a $29, 511 annual profit; and a 60% survival rate will cause a $111, 542 annual profit.
Clearly soft-shell crayfish production is relatively unique because of its potential to realize profit from the first year, in conjunction
with a minimal land requirement, and the opportunity to dual market a product as both bait and food. However, there
are caveats that determine its feasibility; for example, this business requires a well-trained labor force which usually favors
producers with adequate family labor as opposed to hired labor, this business requires constant management during a 120 day
production cycle which makes it difficult for part-time farmers, and this business is dependent on a steady supply of starter
crayfish that is not always reliable because of the lack of aquaculture production or regulatory issues that might reduce the effectiveness
of trapping wild crayfish
4

4
LIMITATIONS TO NUTRIENT REMOVAL IN A FLOW-THROUGH AQUAPONICS SYSTEM
Karen M. Buzby*, Todd P. West, Xinchao Wei and Kenneth J. Semmens
Department of Civil and Environmental Engineering
West Virginia University
Morgantown, WV 26506 USA
kmbuzby@mail.wvu.edu
Aquaponics, the simultaneous culture of fish and plants, can increase profits due to crop diversification as well as mitigate
environmental impacts through reductions in nutrient release. In recirculating fish production systems, aquaponics can reduce
nitrate concentrations by greater than 90% while phosphate concentrations can be reduced 50 - 99% depending on the system.
Aquaponics used in conjunction with flow-through fish production systems, such as those used for trout production, have much
lower nutrient removal rates averaging 12-15% for nitrate and phosphate.
Greater volumes of effluent are generated in flow through systems. In addition, the effluent had colder water temperatures and
lower nutrient concentrations which may affect plant nutrient uptake. An experiment examined the effects of increased water
temperatures, increased nutrient concentrations, increased contact time between plants, and effluent and supplemental light in
an effort to improve nutrient removal. The fish raised were brook trout while lettuce and watercress were used as the plant
component.
Figure 1. Nitrate removal by watercress and lettuce

ADVANCES IN THE CULTURE OF THE LANE SNAPPER Lutjanus synagris
Carina Farías, Jesús Rosas, Aide Velasquez, Jesús León and Tomás Cabrera*
ECAM, Universidad de Oriente
Porlamar 788
Nueva Esparta, Venezuela.
tom3171@telcel.net.ve
Fish culture is an activity under continuous expansion and growth in the Caribbean and tropical areas of the Americas. The fish
family Lutjanidae (snappers) comprises several species that have considerable potential for commercial aquaculture. Among
these species, the lane snapper, (Lutjanus synagris) has shown much potential in preliminary trials. It has an attractive market
demand, and the its relatively small size at sexual maturity is easy to handle during broodstock management. Preliminary
spawning induction and larvae rearing trials were recently conducted for this species at the Institute for Scientific Research,
University of Oriente (IIC/UDO) in Venezuela.
In the spawning trials conducted, ten females with an average weight of 478 g and males with average weight of 518 g. were
used. Fish were maintained in 1 m 3 tanks at a ratio of 2:1 male:female. Water temperature was 58 – 29 o C and salinity was 37
ppt.
Human Chorionic Gonadotropin (HCG) by injection was used for induction, at 15 µg/kg of fish for females and 7.5 µg/kg of
fish for males. After hatching, the larvae were maintained in six tanks using the green-water technique (200,000 cells/ml) with
Isochrysis galbana, Chlorela sp. and Tetraselmis chuii each of them in two different tanks. Rotifers (Brachionus plicatilis)
and Artemia were used as live feeds.
Broodstock fish began spawning 9 hours 30 minutes after the HCG induction. Fertilization rate varied between 87 and 91%,
and the hatching rate 24 hours later was was 66%. Larvae were maintained at a density of 30 individuals/liter in six 400-l tanks,
with filtered seawater at 28 – 29 o C and 37 ppt. A mass mortality was observed after 96 hours in the larvae from the tanks with
T. chuii, and at 120 hours from the tanks with Chlorella sp. The survival rate of the larvae from the tanks maintained with I.
galbana was 1.2% after 15 days, reaching a size of 6.81 mm.
The main problem detected appeared to be related to nutrition and the use of T. chuii and Chlorella sp. in tank rearing water.
Similar problems have previously been detected in the rearing of this species using Chaetoceros ceratosporum at the IIC/UDO
in Venezuela. Further research needs as well as the prospects for development of commercial-scale snapper culture operations
in Venezuela, an area of high priority for the public and private sectors, are discussed.
4

4
DEVELOPMENT AND CHARACTERIZATION OF RIFAMPICIN RESISTANT Flavobacterium
psychrophilum STRAINS AND THEIR POTENTIAL AS LIVE ATTENUATED VACCINE
CANDIDATES
Kenneth D. Cain*, Benjamin R. LaFrentz, Scott E. LaPatra and Douglas R. Call
Department of Fish and Wildlife Resources
and the Aquaculture Research Institute
University of Idaho
P.O. Box 441136
Moscow, ID 83844-1136 USA
kcain@uidaho.edu
Two rifampicin resistant strains of Flavobacterium psychrophilum, 259-93A.16 and 259-93B.17, were generated by routine
passage on TYES plates containing increasing concentrations of rifampicin. Electrophoretic analysis of whole-cell lysates
prepared from the parent and resistant strains identified numerous differences between the 259-93B.17 strain and parent strain,
while there were no differences identified between the 259-93A.17 and parent strains. The LPS banding patterns were identical
between all three strains.
Bacterial challenges of rainbow trout (Oncorhynchus mykiss Walbaum) with the resistant strains demonstrated attenuation of
the 259-93B.17 strain and reduced virulence of the 259-93A.16 strain at the challenge doses tested (Figure 1). Immunization
of rainbow trout with the attenuated live 259-93B.17 strain by intraperitoneal injection (Figure 2) and immersion resulted in
significant protection against challenge with the virulent parent F. psychrophilum strain. Fish were also found to have elevated
specific antibody titers at the time of challenge (data not shown). The results demonstrate that the attenuated 259-93B.17 strain
may serve as an effective live vaccine for the prevention of F. psychrophilum infections.
Figure 1. Virulence comparison of attenuated and wild
type Flavobacterium psychrophilum strains.
Figure 2. Protection observed following immunization
with the attenuated 259-93B.17 strain.

DEVELOPMENT OF INTENSIVE CULTURE METHODS FOR BURBOT Lota lota
Kenneth D Cain, Nathan Jensen, Susan Ireland, John Siple and Matt Neufeld
Department of Fish and Wildlife Resources
and the Aquaculture Research Institute
University of Idaho
P.O. Box 441136
Moscow, ID 83844-1136 USA
kcain@uidaho.edu
Burbot Lota lota are freshwater cod native to Idaho, USA and near demographic extinction from Idaho’s Kootenai River and
British Columbia’s Kootenay River/Lake. Idaho burbot were denied federal listing in 2000 and remain a species of concern.
The Kootenai Tribe of Idaho, the University of Idaho and the British Columbia Ministry of Environment collaborated ca. 2003
and brought wild adult burbot into captivity to develop suitable rearing systems and fundamental hatchery methods as an option
should hatchery burbot be needed to replenish the Kootenai(y) stocks. Over the last five years, observational studies into
adult gender segregation and hormone analog (sGnRha) use have occurred in an attempt to synchronize and control volitional
spawning. An optimal egg incubator design was determined and larval weaning from live diets to commercial larval diets was
evaluated from 2004-2006. Since 2003, facility renovations have occurred annually. The current burbot culture systems in use
consists of full or partial recirculation water systems designed to conserve water and maintain critical water temperatures for
spawning (2-5˚C), egg incubation (3-5˚C), larval feeding and juvenile grow-out (8-20˚C). Additionally, six 8000L fiberglass
tanks are kept outdoors for semi-intensive pond style rearing. Results of spawning observations revealed that volitional (in
tank) spawning is common and fine mesh screens (0.5mm) are needed to keep eggs (typically 1 mm in diameter) within adult
spawning tanks. The optimal egg incubator design is 1L conical bottom upwelling incubators suspended overtop a screened
(0.5mm) 1m circular tank where larvae hatch and collect, develop and begin feeding on live prey. Live prey feeding begins with
brackish (10ppt NaCl) rotifers Brachionus plicatilis mass produced in closed recirculation systems and is followed by Artemia
hatched in 19L water containers (5ppt NaCl). When rotifer feeding ends commercial larval weaning diet (200-600 micron)
feeding begins. Weaning larval burbot to commercial diets, while keeping the rearing environment clean, is the foremost bottle
neck to successful production of healthy juveniles. Therefore, semi-intensive extensive rearing methods are being developed.
All burbot culture system designs and method developments discussed will be applied to create a burbot hatchery manual and
used to design a future conservation breeding program facility aimed at revitalizing burbot populations in Idaho’s Kootenai
River and British Columbia’s Kootenay River/Lake. Such techniques have implications for recovery of burbot populations
elsewhere in the world and have the potential for future commercial aquaculture development of this species.
4

0
SHRIMP AQUACULTURE IN CHINA-CASE STUDY IN HAINAN PROVINCE
Ling Cao* and James S Diana
School of Natural Resources and Environment,
University of Michigan
Dana Building, 440 Church Street
Ann Arbor, MI 48109-1041 USA
caoling@umich.edu
China has become world’s second largest shrimp producers since 1988. Chinese shrimp farms are distributed along almost
18,000 km of coastline from Hainan Province in the tropics to Liaoning Province in the temperate zone. There are about 100
penaeid shrimp species in China’s coastal waters. Main shrimp types cultured in China are Penaeus chinensis, P. monodon,
P. japonicus, P. merguinsis, P. penicillatus, Metapenaeus ensis and P. vannamei, which were introduced from outside of China,
such as Hawaii, USA. Hainan province (Hainan Island), located in the southern part of China, is the major shrimp farming area
in China. From 1995 to 2006, total shrimp production increases from 3,698 to 123,000 metric tones. Although the industry
suffered disease outbreaks and environmental problems, shrimp farming is still expanding rapidly. This study provides background
study of shrimp aquaculture in Hainan Province, China. The objectives are characterizing the operating and management
from hatchery-shrimp farms-shrimp processing plants-market. Problems related to current shrimp aquaculture are addressed.
Emphasis is placed on the shrimp farming impact on the environment. The ecological footprint of shrimp aquaculture
is investigated through life cycle assessment.
Yu, 2007).
TABLE 1. Historical shrimp aquaculture in Hainan Province, China (Hainan Ocean
Fisheries Department, 2008)
Year 1995 2000 2001 2002 2003 2006
Culture areas (km 2 ) 29.4 77.6 92.0 99.3 101.0 119.3
Production
tones)
(metric 3,698 23,250 48,291 67,910 84,016 123,000
FIGURE 1. Cultured brackish water shrimp production in Thailand, China, Indonesia,
Taiwan province of China, Ecuador and globally from 1970 to 2000 (Xie &

RECIRCULATING AQUACULTURE SYSTEM FOR PILOT COMMERCIAL PRODUCTION
OF MARINE FINFISH IN NORTH CAROLINA: PRIVATE-PUBLIC PARTNERSHIP FOR
RESEARCH, EDUCATION AND TECHNOLOGY TRANSFER
Patrick M. Carroll*, Wade O. Watanabe, Thomas M. Losordo, Dennis P. DeLong, J.P. McCann, Glenn Hargett
and Shawn Longfellow
University of North Carolina Wilmington (UNCW)
Center for Marine Science
601 South College Rd.
Wilmington, NC 28403-5927 USA
carrollp@uncw.edu
An environmental education center (Riverworks at Sturgeon City) is under development in Jacksonville, NC. The center’s mission
is the adaptive reuse of a former waste water treatment plant that had discharged into Wilson Bay for more than 40 years
as a headquarters for restoring that habitat and educating citizens to avoid environmental mistakes made in the past. UNCW is
collaborating with the city of Jacksonville to retrofit facilities associated with this wastewater treatment plant for a pilot-scale
recirculating aquaculture system (RAS) for marine finfish and to demonstrate activities that link economic development with
environmental conservation and restoration. Under joint support by MARBIONC (Marine Biotechnology in North Carolina)
and the City of Jacksonville, a pilot RAS was designed by North Carolina State University (NCSU) and UNCW and assembled
on site. The RAS system consists of six 4.62 m-dia. fish culture tanks supported by state-of-the-art RAS components, including
a microscreen drum filter for solids removal, moving bed biofilters for conversion of ammonia to nitrate-nitrogen, foam fractionators
for removal of fine and dissolved organic particulates, ultraviolet sterilizers for disinfection of water, and a geotextile
tube system (“geotube”) for sequestration and management of solid wastes. The RAS is enclosed in an agricultural shelter and
is designed to conduct controlled, replicated studies on the production of high value marine finfish species such as southern
flounder (Paralichthys lethostigma) and black sea bass (Centropristis striata) at high stocking densities and to test economic
viability. In collaboration with a commercial partner (Blue Ocean Farms, LLC), a pilot commercial demonstration project was
recently initiated on site. The commercial partner is operating the pilot RAS as a business, while receiving hands-on training in
cooperation with a multi-disciplinary team from UNCW and NCSU. The project involves undergraduate and graduate students
from local colleges in fish production activities and integrates UNCW’s goals of research, education and technology transfer
through direct cooperation with the commercial end user. A unique aspect of this near commercial scale RAS is that it will
integrate the culture of marine finfish with the culture of microalgae and bivalves in municipal waste drying beds adjacent to the
fish barn. Research will evaluate the use of nutrient-laden effluent from the RAS as a nutritive base to produce microalgae (e.g.
Chaetocerous gracilis), which will then be used to support the substantial feed demands of marine bivalves, e.g. Eastern oyster
(Crassostrea virginica). Oysters produced in this polyculture system can then be utilized for outplanting into Wilson Bay
where they may continue to reduce nutrient levels and improve water quality. Converting expensive nitrogen and phosphorus
into a valuable product will increase profit for the fish farmer, diversify production and reduce pollution.
1

2
PHOTOPERIOD, AGGRESSIVENESS AND ANDROGENS IN CICHLID FISH Tilapia rendalli
Thaís B. Carvalho*, Eliane Gonçalves-de-Freitas and Rui F. Oliveira
UNESP, Universidade Estadual Paulista
IBILCE, São José do Rio Preto, Brasil
thaisbillalba@gmail.com
Photoperiodic manipulation is emerging as an
acceptable approach of practical application for
increasing growth and reproduction in fish farming.
However, light/dark cycles also can increase
aggressive behavior and androgen levels in fish.
This high aggressiveness is harmful in aquaculture
systems, because it can increases wounds and
mortality of fish.
We tested the effect of photoperiod in aggressive
behavior and the plasma androgen levels (testosterone
and 11-ketotestosterone) in adult males of Tilapia
rendalli. We compared two photoperiods: longday
(16L:08D) and short-day (08L:16D).
Animals were grouped for 20 days in each photoperiod.
After that, animals were separated in two
conditions. Condition I: animals were isolated for 4
days and the basal androgen levels was determined
(long-day: n=14; short-day: n=9); Condition II:
animals were isolated for 3 days and in 4 th day they
were submited to territorial intrusion for 1 hour.
The androgen levels in social challenge was given
(long-day: n=12; short-day: n=10).
The aggressive behavior of winner, loser and pair
were stimulated for long-day (Figure 1). The testosterone
and 11-ketotestosterone levels were similar
between two photoperiods. The androgens levels
were not correlated with aggressive interactions.
Thus, photoperiod interfere in social interactions
in T. rendalli, but this result can not be modulated
by androgens. Moreover, the effect of light phase
in aggressiveness shows that artificial photoperiod
manipulations in fish farming must be made with
caution, because light can modify fish behavior.

EVALUATION OF LARVAL FLORIDA POMPANO Trachinotus carolinus FED THE
CALANOID COPEPOD Pseudodiaptomus pelagicus
Eric J. Cassiano*, Cortney L. Ohs, Charles R. Weirich, B. Denise Petty and Jeffrey E. Hill
University of Florida
School of Forest Resources and Conservation
Program in Fisheries and Aquatic Sciences
Indian River Research and Education Center
2199 South Rock Road
Ft. Pierce, FL 34945 USA
ericcass@ufl.edu
The Florida pompano (Trachinotus carolinus) is a high-value marine fish species which plays an important role in recreational
and commercial fisheries of the south Atlantic and gulf coasts of the United States. The culture of Florida pompano (Trachinotus
carolinus) larvae has recently been refined and there is growing interest in its production.
One of the major bottlenecks to marine fish production, including Florida pompano, has been low survival during the larval
phase. Rotifers (Brachionus spp.) and Artemia spp., the traditional live feeds during this phase, are not nutritionally complete
and require use of enrichments to achieve acceptable survival. Recently, the use of copepods as a primary or supplemental live
feed has been evaluated with many species and shown to greatly increase growth, survival, and resistance to stress. Copepods
are the natural food of larval fish species in the wild, have a high nutritional value without enrichment, and show promise in
their ability to be cultured for mass production. By instituting copepod nauplii as a food source during this first feeding phase;
the survival, growth, and subsequent condition of Florida pompano larvae should be improved.
Three 9 day rearing trials were conducted to evaluate feeding regimes in 12 L, 50 L, and 110 L tanks. The feeding of rotifers
(Brachionus sp.), copepod nauplii, mixed diets of each, and a mesocosm of Pseudodiaptomus pelagicus were evaluated. In
each trial, four replicate tanks were used for each treatment. Fish larvae were stocked at a rate of 50/L and fed four times daily
at a rate of 2.5 individuals / mL / feeding from day 2 to day 9 post hatch. Water quality parameters and culture conditions were
monitored daily. Percent survival, growth, and stress resistance were compared for all treatments. Survival was assessed at 9
days post hatch and morphometric data was calculated from samples taken at 0, 3, 6, and 9 days post hatch using image analysis
software.
3

4
DETERMINING RIPENESS IN WHITE STURGEON FEMALES: A NONLETHAL METHOD
TO MAXIMIZE YIELD AND QUALITY OF CAVIAR
Anna G. Cavinato and Sarah A. Servid
Department of Chemistry and Biochemistry
Eastern Oregon University
One University Blvd.
La Grande, OR 97850 USA
acavinat@eou.edu,
The long-term goal of this project is to optimize caviar yield and quality from white sturgeon (Acipenser transmontanus) at
harvest. Currently, the only means to assess ripeness and properly time harvest of caviar is measurement of oocyte polarization
index (PI) which requires surgical biopsy.
We report a rapid and non invasive method based on short wavelength near infrared spectroscopy (SW-NIR) that has the ability
to detect the onset of atresia and could possibly replace oocyte PI by providing a less invasive and more reliable predictor of
maturity.
In September 2007 15 late-vitellogenic females were sampled and moved from a warm water site to a cold water site. Fish
were serially sampled in November, January and March. Atresia was induced by moving these females back to warm water.
Abdominal scans were collected non invasively by SW-NIR on anesthetized females using a diffuse reflectance fiber optic
probe. Approximately 200-300 ovarian follicles were then removed by catheter via a 1 cm incision in the abdomen and further
scanned by SW-NIR. Spectral data were analyzed by Soft Independent Modeling of Class Analogy (SIMCA). Best prediction
of onset of atresia through abdominal scans was achieved by building a model on mature fish with 66% of atretic fish correctly
classified. Best prediction of onset of atresia through direct scans of egg follicles was achieved by building a model on atretic
eggs with 80% of atretic eggs correctly identified. Further studies are currently being performed on a larger fish population to
test the predictive ability of these models on new individuals.

DEVELOPMENT OF A SELECTION INDEX ON BLUE CATFISH FOR OPTIMIZED SPERM
PRODUCTION IN THE CHANNEL X BLUE HYBRID CATFISH PRODUCTION
Nagaraj G. Chatakondi* and Kenneth B. Davis
Eagle Aquaculture, Inc.
339 Aqui Lane
Pike Road, AL 36064 USA
nchatakondi@eagleaqua.com
Channel x blue hybrid catfish is a phenomenal fish for pond culture. This fish represents a major improvement in catfish production
efficiency due to higher growth rates, lower feed conversion efficiency and improved disease resistance and processing
yield compared to the commonly raised channel catfish.
Paucity of hybrid fry in commercial quantities is a limiting factor for the large scale adoption of hybrid catfish in the industry.
Improvements in hybrid fry production has been made through nutritional preparation, temperature manipulation, spawning
protocols and hatchery management addressing the female parent, channel catfish in producing over 90 million fish in 2006-
2008.
Little or no research is focused on the male parent (blue catfish) even though both the gametes are needed for successful fry
production.
Sperm quality is a measure of the ability of sperm to successfully fertilize an egg. Sperm quality in farmed fish may be affected
by different components of bloodstock husbandry, genetics, collection and storage of sperm, fertilization procedures and
hatching protocols. This study proposes to determine the predictive factors for optimum sperm production in blue catfish in
the production of hybrid catfish.
In 2008 spawning season, 139 mature blue catfish were randomly selected representing the four strains and varying age groups
(4 – 10 years). Each male was staged to determine the gonadal maturity, weight (kg), length (cm), plasma testosterone (ng) and
sperm production (10 8 )/kg of body weight (Figure 1).
The maturity of blue catfish exhibited a linear relationship with the sperm production of blue catfish male (y=1.971x – 1.915,
r2=0.8371). Three independent factors : condition factor, plasma testosterone and maturity were regressed to predict sperm
production in blue catfish (r 2 = 0.61). This index will be used to improve the efficiency of the hybrid catfish production and
further to develop methods to raise blue catfish for improved sperm production.

CHANNEL CATFISH FRY PRODUCTION : HATCHERY SPAWNING REPLACE POND
SPAWNING?
Nagaraj G. Chatakondi
Eagle Aquaculture, Inc.
339 Aqui Lane
Pike Road, AL 36064 USA
nagaraj.chatakondi@eagleaqua.com
US farm raised channel catfish Ictalurus punctatus, production relies almost entirely upon natural pond spawning, producing
approximately 2 billion catfish fry annually. Catfish spawning is unpredictable and often varies between 10 – 80% depending
on broodstock, farm management, and environmental factors. The unpredictable and wide variation in reproductive
performance is a limiting factor in the fingerling production under natural pond spawning conditions. The inability to identify
spawning and non-spawning broodfish has hindered selection for reproductive success in genetic improvement programs in
channel catfish.
Superiority of hybrid catfish compared to the commonly grown channel catfish in research and commercial ponds in this decade
will be presented. Higher growth rate, survival and processing yield coupled with lower feed conversion of hybrid catfish
results in lower cost of production.
However, producing commercial quantities of this superior genotype in the last 35 years has hindered its large scale application
in the catfish industry. Recent advances in broodstock preparation and technological innovations in hormone induced spawning
of channel catfish to produce Channel X Blue hybrid catfish production in commercial quantities is presently feasible
(Table1.)
The potential of hatchery spawning to replace the traditional natural pond spawning of channel catfish will be highlighted.
Strategies to produce predictable and efficient fry production under commercial conditions will be discussed.

EXPERIMENTAL USE OF ISOMETAMIDIUM CHLORIDE HYDROCHLORIDE
(TRYPAMIDIUM 1 ) TO CONTROL MORTALITY OF ADULT CHINOOK SALMON
Oncorhynchus tshawytscha DUE TO Cryptobia salmositica
Martin F. Chen*, Henry Y.W. Cheng, Denis Popochock, Brian Russell, Jim Bertolini and Jan Gleckler.
Washington Dept. Fish and Wildlife
600 Capitol Way North
Olympia, WA 98501 USA
chenmfc@dfw.wa.gov
Cryptobia salmositica is a protozoan parasite of blood that infects all five Pacific salmon species and rainbow trout. At Solduc
Hatchery in Washington, prespawning losses of adult spring chinook salmon due to cryptobia have been 50-60% annually for
most of the preceding two decades. Isometamidium chloride (IMC) was injected into adult spring chinook salmon at Solduc
Hatchery in 2007 to control mortality due to Cryptobia. In May 2007, 29 adult salmon were injected (dorsal sinus) twice with 1
mg/kg IMC at 5 week intervals with equivalent controls. Survival to spawning in September 2007 was 38% in the treated group
vs 21% in controls (significant at 95% cl). Eggs were obtained from 4 treated females but no untreated females survived.
Ten adult salmon were injected in July once with 2 mg/kg with an equal number of controls. Survival to spawning was 8/10
treated vs 7/10 controls. Gamete quality from 2 mg/kg injected adults was equivalent or better than controls, measured in survival
of fertilized eggs and juveniles to the first feeding stage.
For 2008, adults were injected once with 2 mg/kg. As of mid-July 2008, the mortality in IMC treated fish was 11% vs 27% in
sham-injected controls. An INAD exemption (file # 011685) has been received from FDA/CVM which permits the release of
progeny of injected adults into public waters.

REVIEW OF COMPUTER-ASSISTED SPERM ANALYSIS (CASA) IN FISH AND SHELLFISH
Ying Chen*, Huiping Yang and Terrence R. Tiersch
Aquaculture Research Station
Louisiana State University Agricultural Center
2410 Ben Hur Road
Baton Rouge, Louisiana 70820 USA
ychen31@lsu.edu
Gamete quality has become a focus in aquaculture due to its impacts on fertilization success and larval survival. Computerassisted
sperm analysis (CASA), initially developed for mammals, is a rapid quantitative method to assess sperm quality by
recording and measuring velocity and trajectory. To date, CASA has been applied to fish and shellfish for sperm quality analysis
in 26 species in 33 published studies (Table 1). Most studies focused on sperm motility changes after cryopreservation, or after
exposure to certain agents such as toxic chemicals or hormones. The species studied were mostly aquaculture fishes such as
catfish, carp, flounder, and trout. Different from that of mammals, fish sperm usually swim for seconds to minutes after activation,
and motility gradually decreases. Therefore, extreme care should be used when setting up the parameter profiles in CASA
systems, especially the timing for initiation of image capture.
Table 1. Broad categories of application by use of computer-assisted sperm analysis for
estimating sperm quality in fish and shellfish in a total of 33 publications from 1995 to 2008.
Sperm motility after cryopreservation
Sturgeon (1996); Muskellunge (1999); Common carp (2000); African catfish (2001); Atlantic
halibut (2006); Red sea bream (2007)
Sperm motility changes after exposure to toxic chemicals
Metal and Tributyltin: Catfish (1996); Catfish and common carp (2002)
Sewage: Goldfish (2002)
Nitrous oxide: Minnow (1998)
Mercury: Goldfish (2003)
Ultraviolet radiation and oxidation: Rainbow trout (2005)
Cypermethrin: Bloch (2008)
Copper: Blue mussel (2008)
Sperm motility changes after exposure to hormone treatments
GnRH injection: Atlantic halibut (2004); European eel (2005)
Endocrine disruption: Flounder (2004)
Xenoestrogen exposure: Spottail shiners (2004)
Sperm motility characteristics
Velocity and motility: Guppy (2006); Amphioxus (2006); Zebrafish (2007); Mussel (2008)
Monthly or seasonal change: Common carp (1996); Atlantic halibut (2006)
Osmotic shock, different buffer, or pH: Carp (1997); Lake sturgeon (1997);
Three-spined stickleback (2004); European eel (2004); European smelt (2006);
After addition of ovarian fluid: Arctic charr (2005)
Between precocious and non-precocious males: European sea bass (2006)
Sperm enzymology
Transferrin function: Carp (2007)
Metabolic enzyme activity: Bluegill sunfish (2005)

CHARACTERIZATION OF 14kDa APOLIPOPROTEIN IN JAPANESE EEL Anguilla japonica
AND DETERMINATION OF ITS HOMOLOGY WITH MAMMALIAN APOLIPOPROTEIN
A-II
Malay Choudhury* and Seiichi Ando
Science of Marine Resources
United Graduate School of Agricultural Science
Kagoshima University
Korimoto, Kagoshima 890-0065, Japan
malayc@gmail.com
Apolipoproteins are lipid binding proteins synthesized mainly in liver and intestine and play an important role in dietary lipid
transport through the circulatory system. Fish in contrast to mammals, utilize lipids in preference to carbohydrate as their
main source of energy. Japanese eel, Anguilla japonica is a fatty fish and useful animal model for studying lipid transport.
14kDa apolipoprotein is one of major protein component of low density lipoprotein (LDL) and high density lipoprotein (HDL)
responsible for lipid transport in Japanese eel. So the proposed work has been undertaken to study in detail about 14 kDa
apolipoprotein in Anguilla japonica and determination of its homology with mammalian apolipoprotein A-II.
Cultured Japanese eel plasma lipoproteins were separated by sequential ultracentrifugation.Plasma lipoprotein electrophoresis
was conducted by gradient PAGE and was transferred to Sequi-Blot polyvinylidene difluoride membrane. The membrane corresponding
to an apolipoprotein with Mr14kDa was sequenced.cDNA was synthesized from the total RNA isolated from the
livers of Japanese eel by 5’and 3’RACE PCR.
The cDNA contained 930bp nucleotides including 87bp of 5’-untranslated region, 426bp of open reading frame, and 417bp of
3’-untranslated region. The coding region of the sequence was translated into 142 amino acids which included a signal peptide
of 20 amino acids. Mature apo-14kDa sequences from Japanese eel satisfied the common structural features depicted for the
exchangeable apolipoproteins: 33-codon block comprised of 3 units of 11 amino acids, and internal repeats of 11 or 22 amino
acid residues predicted to form amphipathic α-helical secondary structures. Human apoA-II consisted of 33-codon block,
followed by internal repeats 4 and 5, while three more internal repeats 6-8 were found in Japanese eel. Japanese eel mature
apo-14kDa sequences showed 28% identities to human apoA-II.From the phylogenetic tree it showed that apoC-II, apoC-III
and apoA-II diverged from a common ancestor before the fish/vertebrate divergence. Apo-14kDa proteins from 13 fish species
including Japanese eel and rainbow trout fell into one group which was much closer to mammalian apoA-II than apoC-II and
apoC-III. The phylogenetic tree was sufficient to consider fish apo-14kDa to be the homologue of mammalian apoA-II and it
was termed apoA-II-like protein.
59

60
CULTURE OF ALLIGATOR GAR Atractosteus spatula IN RECIRCULATING SYSTEMS
Timothy A. Clay*, Mark D. Suchy, Allyse M. Ferrara, Quenton C. Fontenot and Wendell J. Lorio
Bayousphere Research Laboratory
Nicholls State University
Thibodaux, LA 70310 USA
tclay1@gmail.com
Juvenile alligator gar Atractosteus spatula were exposed for 21 days to four trials: (1) fluctuating ambient temperatures
(28.7±1.5 C) and fed 4% body weight split equally among three daily feedings; (2) controlled average temperature(27.5±0.5
C) and fed 4% body weight split equally among three daily feedings; (3) fluctuating temperatures(28.4±2.6 C) and fed 4% body
weight split between two daily feedings; (4) feed amounts of 8, 4, 2, or 1% body weight per day split equally among three daily
feedings. All gar were fed 2.4mm Aquamax extruded floating pellets (45% protein/16% lipid). Fish were stocked at 0.5 fish/l
in 95 liter circular tanks with 60 liters of water at 4 ppt in either individual or four tank recirculating systems. Total biomass
was not different (α=0.05) between the ambient and fluctuating temperatures or between different feeding frequencies. Fish
fed 8% body weight (36.8 g) were larger than fish fed lower amounts. Survival was not different among any treatments, though
there was a general increase in survival in higher feed tanks. Condition was not significantly different among temperature and
feeding frequency trials. Gar fed 4% feed had the highest condition (0.40) and lowest feed conversion rates (2.15). Increasing
feed amounts resulted in significantly increased total-ammonia-N and decreased DO levels, with gar fed 8% feed experiencing
total ammonia-N (9.31±8.9 mg/l) and DO (3.1±1.6 mg/l). The results of this study suggest juvenile alligator gar should be fed
three times a day at 4% body weight.

EFFECT OF FEED DEPRIVATION AND INSULIN-LIKE GROWTH HORMONE ON INDICES
OF PROTEIN DEGRADATION IN RAINBOW TROUT (Oncorhynchus mykiss)
Beth M. Cleveland*, Gregory M. Weber, Kenneth P. Blemings and Jeffrey T. Silverstein
National Center for Cool and Cold Water Aquaculture
ARS-USDA
11861 Leetown Rd.
Kearneysville, WV 25430 USA
beth.cleveland@ars.usda.gov
Insulin-like growth factor-I (IGF-I) is a hormone that promotes growth by both increasing protein synthesis and decreasing
protein degradation. This study utilizes a comparative slaughter approach to determine the effect of feed deprivation and IGF-I
treatment on weight loss and indices of protein degradation. Knowing how proteolytic pathways are affected by nutrient deprivation
and IGF-I will contribute to the understanding of the impact and regulation these pathways have on protein turnover.
Two yearling fish from each of eight rainbow trout families were stocked into 35 30-gallon tanks that were randomly assigned
to five treatment groups (n=7). One treatment group was harvested on day 1 and fish in 14 tanks were implanted with an osmotic
pump containing either recombinant human IGF-I (25 ug/kg/d) or saline. All fish that received impants (2 groups) and an
additional treatment group of fish were harvested after 14 days of feed deprivation. The final treatment group was harvested after
28 days of feed deprivation. For each tank one fish from each family was kept whole for proximate analysis while liver and
white muscle samples were snap frozen for gene expression analysis and blood was collected for 3-methylhistine detection.
Fish treated with IGF-I lost less weight than untreated fish after 14 days of feed deprivation, however, dry weight and fat loss
were not significantly affected by IGF-I treatment (Table 1). A reduction in the mRNA abundance of genes in the proteosome
and cathepsin pathways was observed in IGF-I treated fish (Table 2), indicating that the IGF-I mediated reduction in weight
loss during starvation may involve regulation of these two pathways. Genes in all proteolytic pathways were affected by the
duration of feed deprivation (Table 2). These data indicate that nutrient availability and IGF-I can regulate specific proteolytic
pathways in rainbow trout.
1

2
MECHANISMS CONTRIBUTING TO FAMILY VARIATIONS IN FEED CONVERSION AND
GROWTH IN RAINBOW TROUT (Oncorhynchus mykiss)
Beth M. Cleveland*, Gregory M. Weber and Jeffrey T. Silverstein
National Center for Cool and Cold Water Aquaculture
ARS-USDA
11861 Leetown Rd.
Kearneysville, WV 25430 USA
beth.cleveland@ars.usda.gov
Feed costs often represent greater than 50% of the operating expenses in aquaculture production. Therefore, the efficiency at
which fish convert feed into biomass directly impacts the profitability of a farm. Understanding the physiological and genetic
mechanisms contributing to family variations in feed efficiency will enhance how we use genetic selection to improve feed
conversion in rainbow trout. The rate of protein turnover is a mechanism that affects nutrient retention and, consequently, the
conversion of feed into biomass. This study investigates how indices of protein degradation correlate to family variations in
feed conversion and growth.
Yearling fish (n=7) from each of eight families were individually stocked into 6- or 9-L tanks in a closed recirculation system.
Water was exchanged twice daily and ammonia, nitrite, and nitrate levels were monitored twice weekly. On day 1 of the
feeding trial fish weights were recorded and for five weeks fish were fed twice daily to satiation. Final weights and total feed
consumption were determined at the conclusion of the trial. Also on day 1, 14 fish from each family were harvested from 7 30gallon
tanks (n=2 fish/family/tank). One fish per family was kept whole for proximate analysis while liver and white muscle
samples were removed from the remaining fish and snap frozen for gene expression analysis.
Family feed conversion ratio was correlated with the mRNA abundance of several proteolytic transcripts in both liver and
muscle. The mRNA abundance of the ubiquitin ligase, MAFbx, was positively correlated with feed conversion ratio (g fed/g
gain) in both liver (P=0.0092) and white muscle (P=0.03) and negatively correlated with body weight in liver (P=0.04). These
data suggest that variations in regulation of the ubiquitin-proteosome pathway can impact feed efficiency and growth. Additionally,
there was a positive trend between liver cathepsin L (P=0.060) and caspase 3 (P=0.058) mRNA abundance and family
feed conversion, indicating that these proteolytic pathways can also impact performance. A significant family effect (P

WRITING “SMART” OBJECTIVES FOR EXTENSION PROGRAM DEVELOPMENT
David Cline
203 Swingle Hall
Auburn University, AL 36849 USA
clinedj@auburn.edu
Many of us are called upon to write objectives for various projects. They are an integral part of effective Extension program
planning and evaluation. Strong objective statements also play a key role in successful grant writing. There are a number
of components in a successful objective statement. A simple acronym used to set objectives is called SMART objectives.
SMART stands for:
1. Specific – Objectives should specify what they want to achieve.
2. Measurable – You should be able to measure whether you are meeting the objectives or not.
3. Achievable - Are the objectives you set, achievable and attainable?
4. Realistic – Can you realistically achieve the objectives with the resources you have?
5. Time – When do you want to achieve the set objectives?
The benefit of going through the SMART objective process is that it helps clarify exactly what you are trying to accomplish and
provides a built-in management tool to measure your success.
3

4
ONLINE SURVEY TOOLS – NEW OPTIONS FOR MARKETING AND PLANNING
David Cline
203 Swingle Hall
Auburn University, AL 36849 USA
clindj@auburn.edu
Surveying was once a complex and expensive proposition for researchers. While it still takes significant skill and knowledge
to design a non-biased and easy to analyze survey the distribution and response collection process has improved a great deal.
It is now possible to create, distribute and analyze small or large surveys efficiently and inexpensively with a variety of on-line
tools and services. There are a number of entities providing these services including Survey Monkey and Zoomerang. We will
briefly examine some of these tools, how they work and how to get started.
DEVELOPING A NICHE MARKETING PLAN: THE 5 P’S
David Cline
203 Swingle Hall
Auburn University, AL 36849 USA
clinedj@auburn.edu
Innovative approaches to marketing are usually the key to financial success or failure of small-scale aquaculture producers.
Regardless of the size or type of venture, marketing is an essential component and requires a plan. Most producers would like
to sell to one or two high-volume buyers such as a processing plant or distributor. This is a good marketing strategy if you are
producing large quantities of fish. However, small-scale producers cannot take advantage of the economies of scale of larger
producers and must therefore sell for a higher price to remain profitable. Their best option is to establish niche markets for
their products. Considerable time is required to analyze and develop niche markets and a number of critical issues should be
examined before marketing begins. These issues include marketing’s 5 P’s of Product, Price, Promotion, Place and Planning.

AQUACULTURE ONLINE: HOW PRODUCTION OF ONLINE VIDEO BASED COURSES
BENEFIT PRODUCERS, STUDENTS & INSTITUTIONS
Nathan J. Cochran*, Charles Weibel and Tod Porter
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
nathan.cochran@kysu.edu
Online courses are becoming commonplace on university campuses, although, at this time, few online aquaculture courses are
available. At Kentucky State University’s Aquaculture Research Center, three aquaculture courses are now offered online:
Principles of Aquaculture, Basics of Fish Diseases and Fish Genetics. Two of the three courses are video based giving the
distance learning student more than text-oriented literature. With the use of video lectures that include PowerPoint illustrations
and recorded laboratory demonstrations, students receive a visual understanding of the principles needed to comprehend the
course content.
Class lectures and laboratory demonstrations are videotaped simultaneously in two formats: DVD and mini DV, and are edited
with Adobe Premier Pro software. PowerPoint slides are converted to JPEG images and placed within the video for better
resolution. After extracting the edited video, classes are converted into Flash format using Sorenson Squeeze software. The
courses are hosted on Blackboard, the online platform for KSU and many other universities. Distance-learning students can
access the course online through this platform, take exams, complete written assignments and communicate with the professor.
The introduction of podcasting allows students to download KSU’s aquaculture classes from iTunes University for viewing on
portable video players such as iPods. This method of viewing courses away from a computer monitor provides extra convenience
for students who do not have access to a PC at their leisure. This frees the student to watch classes when time is most
available, whether they are waiting in an airport, commuting to work or on lunch break.
Online courses give non-traditional students and persons from around the world the chance to take classes that would otherwise
be unavailable to them due to location or work schedules. Over the past three years, students from 31 states and 15 countries
have enrolled in KSU’s online aquaculture courses. With increased gas prices and laborious work schedules, many people,
including students and aquaculture producers, are prevented from attending a class on campus. Online aquaculture courses
allow the same educational experience without the student ever leaving home or work.

GENETIC ANALYSIS OF MICROSATELLITE MARKERS LINKED TO SPAWNING TIME
QTLs IN BROODSTOCKS OF RAINBOW TROUT Oncorhynchus mykiss WITH TWICE-
ANNUALLY SPAWNING TIME
Nelson Colihueque*, Rosy Cárdenas, Lorena Ramírez, Francisco Estay and Cristian Araneda
Departamento de Ciencias Básicas
Universidad de Los Lagos
P.O. Box 933, Osorno, Chile
ncolih@ulagos.cl
Broodstocks with a double annual reproductive cycle (DARC) that determine twice-annually spawnings are present in the rainbow
trout. The genetic basis for this behavior is unknown. Nevertheless, it is presumed that it would be related to the spawning
time character (SPT), which is controlled by quantitative trait loci (QTLs). Several microsatellite markers linked to QTLs
for SPT have been described (Sakamoto et al., 1999; O´Malley et al., 2002). This information is useful in order to know the
existence of QTLs for SPT in different broodstock populations, through marker-character association studies. This study was
undertaken to determine the existence of association between microsatellite markers linked to QTLs for SPT with the DARC
trait, present in female broodstock of rainbow trout population, cultured in southern Chile.
A stock, carrier of the DARC trait (twice-spawner, Whyteville 02 strain), was studied together with another non-carrier stock of
this trait (once-spawner, Steelhead strain). Samples of specimens of these two populations were genotyped with six microsatellite
markers linked to QTLs for SPT, and four that were unlinked. Microsatellites were amplified by PCR and the genotypes
determined by polyacrylamide gel electrophoresis. The interpopulational allelic frequency heterogeneity was determined, as
well as the Choulakian and Mahdi (2000) marker-character association index.
Three microsatellite loci linked to QTLs for SPT (One3ASC, One19ASC and One112ADFG) presented allelic heterogeneity
and a highly significant marker-character association with the DARC character (Table 1). On the other hand, none of the microsatellite
loci that were not linked to QTLs for SPT presented this pattern. The results indicate that the DARC trait in rainbow
trout would be based on the existence of QTLs for SPT. The present study contributes to knowledge of the genetic basis of
the DARC trait in this species and to its potential use in salmon farming. This study was financed by the grant FONDECYT
1060623.

ENERGY USE, RESOURCE CONSUMPTION, AND SUSTAINABILITY OF AQUACULTURE
SYSTEMS – AN INTRODUCTION
John Colt
National Oceanic and Atmospheric Administration
Northwest Fisheries Science Center
2725 Montlake Blvd. East
Seattle, WA 98112 USA
john.colt@noaa.gov
Sustainable development implies that current demands will not compromise the ability of future generations to meet their own
needs and that food production systems should be as efficient as possible and minimize environmental impacts. Different production
systems will have different labor, energy, and physical components. To compare these systems from a sustainability
perspective, it is necessary to be able to evaluate the components of each system in terms of some type of “common currency.”
The four commonest types of sustainability analysis are:
Net Energy Analysis. Energy analysis (or net energy analysis) is a form of energy accounting that considers both direct and
indirect energy inputs to a given process. The major aquaculture inputs are material inputs (feed, pure oxygen, calcium carbonate),
energy inputs (electrical, gasoline/diesel, and natural gas), labor, and gametes. Fixed capital components such as concrete,
steel, aluminum, fiberglass, and plastics in the facility are considered. Each input or output may have three types of energy:
direct energy, indirect energy, and transportation energy. Direct energy is the amount of heat (∆H) that is released if the compound
is burned in a bomb calorimeter. Indirect energy is the amount of energy that was needed to produce a unit weight of a
given compound. The transportation energy is the energy required to transport material to and from the facility. Energy analysis
is based on estimation of energy density (MJ/kg) and the mass of the compound used.
Greenhouse Gas Emissions. Greenhouse gas emissions (carbon dioxide, methane, nitrous oxides, and fluorocarbon) are of
current interest because their potential impact on global warming and ocean acidification. Greenhouse gas emissions are based
an emission factors (kg/kg) and the mass of the compound consumed.
Life Cycle Analysis. Life cycle assessment is a “cradle-to-grave” approach for assessing industrial systems. The impacts are
classified into global (climate change, ocean acidification, ozone depletion, resource depletion), regional (photochemical smog,
acidification), and local (human health, terrestrial toxicity, aquatic toxicity, eutrophication, land use, water use). Because LCA
considers all three media (air, land, and water), it helps to prevent shifting environmental problems from one media to another,
or from one impact category to another. LCA methodology has been standardized by the International Standards Organization
(ISO) 14000 series. The amount of information needed to conduct an LCA is immense and typically requires the purchase of
commercially available LCA software packages.
Ecological Footprint Analysis. Ecological footprint analysis estimates the surface areas of the ocean and land needed to support
a given facility. This included area to produce the feed and process the wastes generated by the facility. Small ecological
footprints are assumed better than large footprints and are typically expressed in terms of the area of the aquaculture facility
(i.e., ha of ocean per ha of facility).

ENERGY AND RESOURCE CONSUMPTION OF LAND-BASED ATLANTIC SALMON
SMOLT HATCHERIES IN THE PACIFIC NORTHWEST (USA)
John Colt * , Steve Summerfelt, Tim Pfeiffer, Sveinung Fivelstad and Michael Rust
National Oceanic and Atmospheric Administration
Northwest Fisheries Science Center
2725 Montlake Blvd. East
Seattle, WA 98112 USA
john.colt@noaa.gov
Conventional economic analysis does not typically include societal costs associated with ecological or environmental impacts
and may significantly underestimate production costs and ecosystem impacts. Different production systems will have different
labor, energy, and physical components. To compare these systems from a sustainability perspective, it is necessary to be able
to evaluate the components of each system in terms of some type of “common currency.”
Two potential analytical tools for sustainability analysis include (a) energy analysis and (b) greenhouse gas emissions. Energy
analysis is a form of energy accounting that considers both direct and indirect energy inputs to a given process. Greenhouse gas
emissions are of current interest because their potential impact on global warming and ocean acidification.
This paper evaluates the resource and energy requirements of six different types of land-based, hatchery production systems
located in the U.S. Pacific Northwest: flow-through with a gravity water supply, flow-through with a pumped water supply,
flow-through with pure oxygen, partial reuse system, partial reuse with heating, and a reuse system for the production of Atlantic
salmon (Salmo salar) smolts. Key parameters used in the evaluation include direct energy, indirect energy, transportation
energy, greenhouse gas emissions, and pollutant discharges.
Power (electricity and natural gas) and feed energy accounted for the majority of the required energy for all the rearing option
evaluated. The sum of the fixed capital and chemicals components accounted for less than 2-12% of the total energy budget for
any rearing option. The energy efficiency (energy output/energy input) of the six options ranges from 0.97% for flow-through
with pumped supply to 3.49% for the flow-through with gravity supply. The rearing options with the three highest energy efficiencies
were flow-through with gravity supply (3.49%), partial reuse (2.75%), and reuse (2.64%).
On a kg of smolt produced basis, the six rearing options showed a wide range in performance. The reuse system had the lowest
water (2 m 3 kg -1 ) and land (0.13 m 2 kg -1 ) requirements and the third lowest total energy requirement (288 MJ kg -1 ). The partial
reuse system had the second lowest total power requirement (276 MJ kg -1 ), a low land requirement (0.21 m 2 kg -1 ), and moderate
water requirements (33 m 3 kg -1 ). The partial reuse with temperature control had the second highest total power requirement (657
MJ kg -1 ) and land and water requirements similar to the partial reuse system without temperature control. The flow-through
system with pumped water supply had the highest water (289 m 3 kg -1 ), land (2.19 m 2 kg -1 ), and energy requirements (786 MJ
kg -1 ) of any of the rearing options. By comparison, the flow-through system with gravity water supply had the lowest energy
requirement (218 MJ kg -1 ), a moderate land requirement (0.78 m 2 kg -1 ), and a high water requirement (214 m 3 kg -1 ). The ranking
of the six rearing options based capital and operating costs are likely to be quite different from those based on energy, water,
and greenhouse gas emissions.

COMPARISON OF ENERGY AND RESOURCE CONSUMPTION OF COMMERCIAL SIZED
ATLANTIC SALMON PRODUCTION SYSTEMS IN BRITISH COLUMBIA, CANADA
John Colt
National Oceanic and Atmospheric Administration
Northwest Fisheries Science Center
2725 Montlake Blvd. East
Seattle, WA 98112 USA
john.colt@noaa.gov
There is increased interest in closed containment production systems that may have reduced impacts on wild stocks and the
overall marine ecosystem. While closed containment systems are thought to have desirable characteristics, their overall energy
and resource consumption is poorly known, especially in relationship to conventional netpen culture. This talk presents the
resource and energy requirements of five types of marine culture systems: (a) conventional netpen, (b) floating closed containment
systems with rigid walls, (c) floating closed containment systems with flexible walls, (d) land-based flow-through system,
and (e) a land-based complex reuse system.
The production capacity of the systems was assumed to be 2,500 tonnes/cycle for all systems. The complex reuse system was
stocked with smolts three times a year; all other systems were stocked once and all the fish harvested at the end of the production
cycle followed by a 60 day fallow period. The energy use of these ten culture options was evaluated in terms of direct, indirect,
and transportation energy. The energy required for feeding, maintenance, pumping, water treatment, aeration, temperature
adjustment, and transportation was estimated. The contribution of fixed capital was allocated to each option based on the mass
of steel, aluminum, plastics, and wood contained, expected life, and energy density values. The mass of greenhouse gases was
estimated from the energy consumption for each component and greenhouse emission factors. Key performance parameters
included (a) energy consumption, (b) greenhouse gas emissions, (c) discharge of solids and nutrients, (d) water usage, (e) water
and land area needed, and (f) overall energy efficiency.
For a single farm, the total energy used ranged from 199 to 1,576 TJ/cycle. The total greenhouse gas emission varied from
11,000 to 104,000 tonnes/cycle. The amount of solids/cycle varied from 57,000 kg for the complex reuse system to 1,067,000
for the conventional netpen.
In terms of energy efficiency, the three best performing systems were the conventional netpen (9.13%), the closed containment
system with flexible walls, pure oxygen aeration (8.34%), and the closed containment system with rigid walls, pure oxygen
aeration (7.03%). The land-based system with influent aeration and on grade had the lowest energy efficiency (1.22%).
The performance of the best system resulted from low power requirements for life support and waste treatment. Based on
the discharge of solids and nutrients, the complex reuse system was superior to all the other systems, but ranked 7 th in terms
of energy efficiency. The greenhouse gas emissions performance tracked the energy efficiency rankings. The selection of the
“best” system may require trade-offs between the different performance measures selected for this analysis. The ranking of the
ten rearing options based on capital and operating costs are likely to be quite different from those based on energy, water, and
greenhouse gas emissions. In addition, environmental risks, social/economic impacts, and environmental justice issues have
not considered in this analysis.

0
IMPACT OF ATTACHED ALGAE, SUSPENDED BACTERIA, AND RE-AERATION ON
OXYGEN AND CARBON DIOIXIDE BALANCES IN SERIES RACEWAYS
John Colt, Barnaby Watten and Michael Rust
National Oceanic and Atmospheric Administration
Northwest Fisheries Science Center
2725 Montlake Blvd. East
Seattle, WA 98112 USA
john.colt@noaa.gov
In the design of raceways, the impact of algae, bacteria, and oxygen transfer is commonly ignored. In lightly loaded single-pass
salmon and trout systems, these processes may not important. Their impact has not been clearly documented in heavy loaded
serial reuse systems that are used in commercial facilities.
The impact of these parameters were evaluated in a 10-pass serial reuse system operated at a cumulative oxygen consumption
of 16.7 mg/L, elevation = 1,500 m, water temperature = 10°C, and water flow = 2,500 lpm. Oxygen consumption of suspended
bacteria was based on a kinetic constant (K 20C ) of 0.10 and a theta value equal to 1.135. The influent BOD 5 was assumed equal
to 2 mg/L and the BOD 5 production rate equal to 50 g/kg feed. The oxygen production and respiration rate of attached algae
were assumed to be 3.0 g oxygen/(m 2 •d) and 0.103 g carbon dioxide/(m 2 •d), respectively. The reaeration coefficient for oxygen
was based on the O’Connor-Dobbins film penetration model. The reaeration coefficients ratio for CO 2 /O 2 was equal to 0.81. A
triangular notch weir was used between raceways with a r 20 = 1.923.
The impact of attached algae, suspended bacteria, re-aeration, and weir aeration on oxygen and carbon dioxide balances are
presented below for the effluent from raceways 1 and 10 for both day and night conditions: Compared to gas transfer across the
air-water interface, the weir is a much more significant mechanism for gas transfer in raceways. During the day, attached algae
is a significant source of oxygen but has little impact at night. Attached algae significantly reduces the CO 2 concentrations during
the day, resulting in reduced metabolic pH depression and elevated un-ionized ammonia concentrations. Field testing and
verification is needed to confirm these findings.
Oxygen Balance in Raceways 1 and 10
Time #
Influent
DO
Fish Algae Bacteria
Re-
Aeration
Effluent
DO
Weir
To Next
Raceway
Day 1 9.456 -1.667 0.664 -0.0185 0.016 8.450 0.508 8.957
10 8.307 -1.667 0.664 -0.0330 0.051 7.332 0.980 8.302
Night 1 9.456 -1.667 -0.0084 -0.0185 0.026 7.855 0.785 8.573
10 7.432 -1.667 -0.0084 -0.0330 0.088 5.812 1.612 7.424
Carbon Dioxide Balance in Raceways 1 and 10
Time #
Influent
CO2 Fish Algae Bacteria
Re-
Aeration
Effluent
CO2 Weir
To Next
Raceway
Day 1 0.746 2.292 -0.9125 0.0254 -0.008 1.407 -0.177 1.285
10 3.479 2.292 -0.9125 0.0454 -0.082 4.504 -1.025 3.539
Night 1 0.746 2.292 0.0116 0.0254 -0.017 2.127 -0.374 1.818
10 5.634 2.292 0.0116 0.0454 -0.147 7.534 -1.856 5.723

DEVELOPMENT OF STANDARDS FOR FISHERIES AND AQUACULTURE – AN UPDATE
ON THE INTERNATIONAL STANDARD ORGANIZATION –TECHNICAL COMMITTEE 234
John Colt and Scott Cedarquist
Northwest Fisheries Science Center
National Oceanic and Atmospheric Administration
2725 Montlake Blvd. East
Seattle, WA 98112 USA
John.colt@noaa.gov
The International Standard Organization (ISO) develops international standards for products, services, processes, materials and
systems, and for conformity assessment, managerial and organization practices. The ISO organization consists of a network
of national standards bodies from all regions of the world, working in partnership with international organizations such as the
United Nations and the World Trade Organization. The U.S. is represented in ISO by the American National Standards Institute
(ANSI).
On February 2007, the ISO Technical Management Board passed a resolution to approve the formation of the new ISO Technical
Committee 234 (TC234) on Fisheries and Aquaculture and to assign the international secretariat responsibilities of this
TC to Norway. ANSI has delegated the administration of this technical committee to the American Society of Agricultural and
Biological Engineers (ASABE) because of its extensive experience in agricultural standards development.
The plenary meeting of TC234 occurred October 9-10, 2008 in Bergen, Norway. The U.S. delegation consisted of John Colt
(NOAA; head of delegation), Melvin Myers (consultant), Robert McKnight (University of Kentucky), Scott Cedarquist (Director
of Standards, ASABE), and Barbara Montwill (FDA).
TC 234 is organized into participating countries (P countries), observing countries (O countries), and liaisons with international
organizations. Only P members vote on Technical Committee actions. The current participating countries are: Canada, Fiji,
France, Iceland, India, Malaysia, New Zealand, South Africa, Spain, Thailand, USA, United Kingdom, Viet Nam, Sweden,
and Norway. The observing countries are: Argentina, Belgium, Bosnia and Herzegovina, Brazil, Croatia, Cyprus, Denmark,
Finland, Germany, Israel, Italy, Japan, Malta, Montenegro, The Netherlands, Poland, and Ukraine.
In June 2008, the TC234 Secretariat proposed four new work item proposals (NWIP):
• Traceability of fishery products - Specification on the information to be recorded in captured fish distribution chains.
• Traceability of fishery products - Specification on the information to be recorded in farmed fish distribution chains.
• Environmental monitoring of marine fish farms.
• Cage fish farms – Requirements for design, dimensioning, production, installation and operation. (N029)
In September 2008, the P countries will have to vote support or oppose the development of each of these potential standards.
For the standard process to proceed, 11 countries must suppose a particular standard. The next TC234 meeting will be held
November 11-12, 2008 in Madrid, Spain.
1

2
ANALYSIS OF REPEATED MEASURES DATA – JUST WHAT ARE MIXED MODELS?
Loveday L. Conquest
University of Washington
School of Aquatic & fishery Sciences
1122 NE Boat St.
Seattle WA 98195-5020 USA
conquest@u.washington.edu
Analysis of repeated measures data is a problem often faced by aquaculture scientists. This occurs when each of several subjects
is followed through time. The subjects could be individual organisms, or Petri dishes, or tanks of fish. There is often more
than one treatment being compared. An incorrect analysis is to consider this as a two-way analysis of variance (ANOVA),
with Treatment as one factor, Time as the second factor, and the number of data points in each Treatment x Time combination
as independent “replicates”, upon which the error term is based. However, a simple two-way ANOVA commits the error of
temporal pseudoreplication, as this approach does not take into account the fact that each subject produces a “time trace”. Any
analysis of repeated measures data should specifically incorporate the notion of the time trace. This is precisely what mixed
effects models do, as they provide a statistical model with a correlation structure. The term “mixed effects models” arises from
the fact that they ultimately combine fixed effects (which influence only the mean of the response variable) and random effects
(which influence only the variance of the response variable).
This session will introduce the audience to the use of mixed effects models in analyzing data that involve time traces, where
temporal autocorrelation from repeated measures on the same individuals is expected to occur.

THE EFFECTS OF STOCKING DENSITY AND ARTEMIA DENSITY ON GROWTH AND
SURVIVAL OF LARVAL LARGEMOUTH BASS
Shawn Coyle, Leigh Anne Bright, Kyle Schneider and James H. Tidwell
Kentucky State University
Aquaculture Research Center
Frankfort, KY 40601 USA
shawn.coyle@kysu.edu
The intensification of larval rearing techniques for largemouth bass (Micropterus salmoides) have been identified as a priority
area for commercialization of the species. A series of experiments investigating techniques required to raise largemouth bass
fry from first feeding until fully habituated to a commercial dry diet have indicated that largemouth bass swim-up fry readily
consume Artemia nauplii. However, in these experiments Artemia was fed to excess and optimal bass stocking and prey densities
were not determined.
Two independent feeding trials were conducted to determine the optimal fry and Artemia densities for the first two weeks of
largemouth bass larviculture. The duration of each trial was 14 days. First day swim-up fry were collected from individual
spawning mats harvested within 24 hours of spawning and incubated individually to ensure cohorts were at least paternal siblings.
Experiment 1 was a 3X3 factorial and evaluated largemouth bass fry (9 mg) at stocking densities of 20, 40 and 80 fry/L
and Artemia nauplii feed rates of 5, 10 and 20 nauplii/ml administered twice daily, while Experiment 2 evaluated fry stocking
densities of 10, 20 and 40 fry/L and Artemia nauplii feed rates of 20, 30 and 40 nauplii/ml administered twice daily.
In Experiment 1, ANOVA on average harvest weight indicated a significant interaction (P

4
SPERM VITRIFICATION OF A LIVE-BEARING FISH, THE GREEN SWORDTAIL
Xiphophorus helleri
Rafael Cuevas-Uribe*, S.P. Leibo and Terrence R. Tiersch
Aquaculture Research Station
Louisiana State University Agricultural Center
Baton Rouge, LA, 70820 USA
ruribe@agcenter.lsu.edu
Vitrification is an alternative method of cryopreservation where the sample (including the extracellular and intracellular
fractions) forms glass (non-crystalline ice). The probability of successful vitrification depends on interactions of three
major factors: sample viscosity, cooling and warming rates, and sample volume. The first step in protocol development is the
selection of a suitable and least toxic vitrification solution. Sperm vitrification is new in fish and there is little knowledge of
cryoprotectants that vitrify at non-toxic concentrations. The goal of the project was to develop streamlined protocols that could
be integrated into a standardized approach for vitrification of aquatic species germplasm. The specific objectives were to:
1) measure acute toxicity of cryoprotectants at varied concentrations; 2) test various solutions for vitrification; 3) evaluate
different warming methods, and 4) evaluate the ability to fertilize eggs.
Sperm were collected from the green swordtail Xiphophorus helleri by crushing of dissected testis and diluting to a final concentration
of 1 x 10 8 cells per ml using Hanks’ balanced salt solution at 500 mOsmol/kg (HBSS 500). The acute toxicity of
11 cryoprotectants, 50 vitrification solutions, and 5 commercial vitrification solutions were tested. For each cryoprotectant,
concentrations were evaluated ranging from 5 to 50% (Figure 1). In Experiment 2, the glass formation of 44 solutions was
evaluated using 5-mm nichrome loops. After plunging the solutions in liquid nitrogen, glass formation was identified by a transparent
state (crystallization produced a milky appearance; Figure 1). In Experiment 3, four different warming solutions (fresh
water, HBSS 300, HBSS 500, and 7% ethylene glycol + 7% 1,2 propanediol) at two temperatures (24 and 37 °C) were tested.
In Experiment 4, 20 females of X. maculatus were artificially inseminated with two replicates of vitrified sperm from each of
10 males. In addition, five albino females of X. helleri were inseminated with vitrified sperm.
From 226 treatments evaluated for acute toxicity, 44 were selected for further testing, and of these, 21 formed glass. The average
motility of sperm in these vitrifying solutions was low (0-20%; control 60%). The best warming solution was found to be
HBSS 300, and there was no difference between 24 °C and 37 °C. The best post-thaw motility (≤ 10%) was found in 40%
glycerol, and in 20% ethylene glycol + 20% glycerol with an exposure time of ≤ 2 min before cooling. For artificial insemination,
the loop was warmed in 5-µl of HBSS 300 at 24 °C. A 5-µl volume was injected into the females. Vitrification can be
simple, fast, and cheap, does not require expensive equipment and can be performed in the field. This is important for fishes
such as Xiphophorus which are valuable biomedical models, yet many are threatened species in the wild.

EVALUATION OF DIFFERENT ORGANIC FERTILIZERS AND STOCKING GRADED
JUVENILES AS MANAGEMENT PRACTICES FOR LOW INPUT PRODUCTION OF
FRESHWATER PRAWNS Macrobrachium rosenbergii IN EARTHEN PONDS
Louis R. D’Abramo* and Terrill R. Hanson
Department of Wildlife and Fisheries
Mississippi State University
Box 9690
Mississippi State University
Ldabramo@cfr.msstate.edu
Juvenile freshwater prawns were stocked at a density of 24,100/ha into 0.05 hectare earthen ponds that two weeks prior had
been fertilized with a one time application of cottonseed meal at 224.2 kg/ha followed by additions of 16.8 kg/ha every other
day. There were five treatments, five replicates (ponds) per treatment. All ponds contained vertically orientated substrate (orange
plastic fencing) supplied at a rate of 25 % of the bottom surface area. One group of three treatments were designed to compare
the efficacy of the different organic fertilizers, range cubes, alfalfa pellets, and soy hull pellets, using ungraded juveniles
with a mean stocking weight of 0.215 g. The effect of stocking ungraded juveniles (mean weight = 0.215 g) versus juveniles
from the same population graded into two groups (mean weight 0.275 g and 0.123 g) was also evaluated using another group
of three treatments in which ponds were fertilized with range cubes. The total amount of organic fertilizer applied to all ponds
for the entire growing season (128 days) was calculated based on a 3:1 ratio of fertilizer to anticipated yield at harvest. Daily
application rates of the organic fertilizers ranged from 13.1 to 33.6 kg/ha/day and were calculated from proportions of the total
amount assigned to different time intervals (weeks) during the growing season. The effects of fertilizer type and grading on
survival, mean harvest weight, and total production, and the results of a corresponding economic analysis will be presented.

USE OF INSECT MEALS AS COMPLETE SUBSTITUTES FOR MENHADEN FISH MEAL IN
DIETS FED TO JUVENILE HYBRID STRIPED BASS
Louis R. D’Abramo* and Ernie Papadoyianis
Department of Wildlife and Fisheries
Mississippi State University
Box 9690
Mississippi State University
Ldabramo@cfr.msstate.edu
The nutritional value of two meals derived from mass rearing of insects was evaluated as a complete substitution for menhaden
fish meal in diets fed to juvenile hybrid striped bass. Six juvenile hybrid striped bass, mean weight ranging from 15.0 to 17.9g,
were stocked into individual aquaria (working volume = 72.5 L) that were part of a flow through system. Water temperature
and dissolved oxygen were recorded twice daily and ranged from 22.5 to 25.3 C and from 5.2 to 9.5 ppm, respectively. There
were five dietary treatments, a control diet containing menhaden fish meal, and four experimental dietary treatments in which
menhaden fish meal was completely substituted with meals from insect A, insect B, a combination of A and B (50:50), or insect
A supplemented with methionine and menhaden fish oil. The amount of insect meal added to each diet was based upon maintaining
the dietary level of crude protein at 44.0 %, and all diets were formulated to have similar EPA:DHA ratios. The percent
composition of crude lipid in the diets ranged from 15.5 to 21.2 %. Fish were fed twice daily to satiation until the experiment
was terminated at 45 days. Fish fed diets containing insect meal grew at a rate of approximately 72 % of those fed the control
(fishmeal) diet. Fish fed the control diet showed a mean percent weight gain of 327.3 % which was significantly higher than
those of fish fed diets containing the insect meals (range = 150.4 to 240.8 %). Mean survival did not significantly differ among
fish fed the different diets and ranged from 95.2 to 100.0 %. Feed conversion ratios among dietary treatments ranged from 1.1
to 1.3 and protein efficiency ratios ranged from 1.68 to 1.98, respectively, with fish fed the control diet showing the highest efficiencies.
The lower growth rates observed for fish fed the insect meal containing diets may be the result of insufficient levels
of HUFAs and the quality and availability of essential amino acids. Results do suggest a high potential for the use of insect
meal as an alternative feedstuff for the aquaculture of carnivorous species of fish, particularly if the quality of lipid in the insect
meal can be modified by the diet used to feed the insects.

RECLAIMED AQUACULTURE EFFLUENT FOR USE ON CUCUMBERS Cucumis sativus cv
Eureka GROWN IN THE U.S. VIRGIN ISLANDS
Jason J. Danaher* and James E. Rakocy
University of the Virgin Islands
Agricultural Experiment Station
RR 1 Box 10,000
Kingshill, VI 00851 USA
Experiments have demonstrated aquaculture effluent to be an excellent nutrient source for agronomic crops; thus, effluents
should be treated as a resource and not as a disposal problem. Technologies now exist to reclaim aquaculture effluent and
separate the solid fraction from the liquid portion; hence, giving more options for land application. A previous study at UVI
found a Geotube ® retained high concentrations of total suspended solids in aquaculture effluent and created filtrate that passed
through T-tape ® irrigation.
The objective of the experiment was to compare reclaimed aquaculture effluent using a Geotube ® with a commercial, inorganic
fertilizer. A Randomized Block Design was used with three treatments and three replicates each. The control used 13-13-13
Osmocote ® and was irrigated with rainwater. Treatment two used retained solids from the Geotube ® and was irrigated with
rainwater (GSRW). Treatment three also used Geotube ® solids, but was irrigated using Geotube ® filtrate (GSGF). Fertilizer
was applied at 107 kg/ha N. Soil analysis was compared at planting and at harvest. Plant tissue analysis was performed before
fruit set. Average fruit weight, total number of fruit, total yield and marketable yield were compared between treatments using
ANOVA.
Results found GSGF and GSRW had significantly higher (P < 0.05) total number of fruits, total yield, and marketable yield
compared to the Control (Table 1). There was no significant difference (P ≥ 0.05) in the average fruit weight between treatments.
There was no significant difference (P ≥ 0.05) in plant tissue analysis between treatments (Table 2); however, plant
tissue nutrient levels were below recommended ranges in all treatments. A second trial will be conducted using incrementally
increased rates of Geotube ® retained solids based on soil analysis and a recommended rate of application.
77

A DISCRETE CHOICE EXPERIMENT TO UNDERSTAND THE PREFERENCES OF
AUSTRALIAN CONSUMERS IN BUYING PRAWNS
Dr Nick Danenberg, Dr Simone Mueller and Dr Hervé Remaud
Ehrenberg-Bass Institute for Marketing Science
University of South Australia
GPO Box 2471
Adelaide – SA – 5001 – Australia
nick.danenberg@unisa.edu.au
A Discrete Choice Experiment was conducted to survey and investigate the preferences of Australian consumers for prawns
(shrimp) in relation to various attributes. The attributes included for analysis were: price ($12.50-$32), region of origin (Australia,
Spencer Gulf (AUS), China and Thailand), method of production (farmed vs. wild caught), method of storage (fresh vs.
frozen), health claims (rich in Omega-3, low in fat, none), and sustainability claim. Respondents were presented choices in
a graphical format using the internet, which very closely simulated the real-life prawn retail purchase environment. A Latent
Class choice analysis was conducted, able to model consumer heterogeneity. This analysis is simultaneously estimating individuals’
attribute level part-worth, the relative importance of the selected product attributes and cluster membership based on
respondents’ preference data.
The study found that at the aggregate level, region of origin is the biggest driver for the purchase of prawns, representing over
60% of the consumer decision making process, followed by price at 20%. At the segment level, however, the cluster analysis
additionally revealed that there were five distinct, substantial groups of consumers. One such segment (30% of consumers)
valued low prices more than region; and another segment (9% of consumers) predominantly valued freshness. Contrary to the
elsewhere reported importance to consumers of the promotion of health benefits and environmental sustainability, this study
revealed that overwhelmingly, consumers did not place much value on either health or sustainability claims.
When trying to understand drivers of consumer purchase, marketers face a critical issue: consumers don’t necessarily do what
they say. Most market research methods employ direct questioning approaches, where consumers are asked (in different ways)
about what products or features they like. Choice Modeling, in particular, Discrete Choice Experiments (DCE), differs from
this in that it analyses consumers’ choices from controlled alternatives and thereby simulates the decision making process that
consumers apply in the real world.
Through having respondents repeatedly make choices, or trade-offs, in selecting one offering from a competitive set of close
alternatives, a DCE allows a researcher to determine the importance of given product attributes (or, features) and the favourability
of different levels on these attributes—e.g., ‘is low in fat, as a level of the attribute health claim favoured over rich in
Omega-3?’. DCEs have proven to provide valid predictions of real world sales of a very diverse range of products and services,
including food products.
We report on a survey conducted in early 2008 that analyzed Australian consumers’ choice behavior for prawns. An experimental
design was employed to construct the choice sets based on the attributes and levels stated above. The survey with 1,276
respondents was administered over the internet, which allowed for a realistic simulation of a characteristic retail environment
for prawn purchasing.
As the DCE reveals the underlying importance of attributes and the preference for attribute levels, these results can be used to
simulate a market with competing products, assuming that these only vary in the attributes controlled in the experiment. Such
a Decision Support System (DSS) predicts market shares of products with different characteristics. For instance, it can forecast
the market share of a regional Australian brand compared to national and imported brands if the producer is accredited ‘sustainable
fishery’ or uses a health claim such as ‘rich in omega-3’, and so on. Such a DSS allows the seafood industry to assess their
strategic options before introducing them into the real market.

THE SYNERGISTIC USE OF DIETARY BIOTIC SUPPLEMENTS FOR ENHANCING THE
GROWTH AND SURVIVAL IN THE LARVAL STAGES OF EUROPEAN LOBSTER Homarus
gammarus
Carly L. Daniels*, Simon J. Davies and Dom. Boothroyd
The University of Plymouth
Devon, PL8 8BL, UK
carly.daniels@nationallobsterhatchery.co.uk
With a growing realisation that prevention is better than cure the use of prebiotics and probiotics, as dietary supplements, is developing
as an essential in aquaculture, world and species wide. We tested the effects of commercial prebiotics and probiotics
on survival, growth performance, post larval quality and feed/cost benefit in Zoeal European Lobster, Homarus gammarus.
Four replicate 30 day feeding trials were conducted at the National Lobster Hatchery, Cornwall, UK. In a single trial four larval
cones (80l) were each stocked with 440-1080 zoeal I lobsters (dependent on replicate) and offered enriched Artemia feed at
5 per ml. Cones were offered with one of four diets containing prebiotic (MOS - Bio-Mos ® ) and/or probiotic (Bacillus spp.
- Sanolife ® ) (Control, Diet 1 (165ppt Bacillus spp.), Diet 2 (20ppt MOS), Diet 3 (165ppt Bacillus spp. and 20ppt MOS)) for 30
days. Five lobsters from each cone were measured (weight and carapace length) on day 1, 3, 9, 13, 18 of development. Moulting
success was also recorded from day 14 to 30 with survival calculated at day 30. After 18 days larval H. gammarus lobsters fed
a diet simultaneously supplemented with prebiotics and probiotics (Diet 3) had significantly (P

0
EVALUATION OF POTASSIUM PERMANGANATE AGAINST AN EXPERIMENTAL
SUBACUTE INFECTION OF Flavobacterium columnare IN CHANNEL CATFISH, Icatlurus
punctatus
Ahmed M. Darwish*, Andrew J. Mitchell and David L. Straus
Harry K. Dupree-Stuttgart National Aquaculture Research Center
Agricultural Research Service
U.S. Department of Agriculture
Stuttgart, Arkansas 72160 USA
Ahmed.Darwish@ars.usda.gov
The efficacy of potassium permanganate (KMnO 4 ) as a prophylactic and therapeutic treatment for subacute infection of Flavobacterium
columnare was demonstrated in experimentally infected channel catfish, Ictalurus punctatus. Catfish experimentally
infected with F. columnare to mimic a subacute infection, were dosed with KMnO 4 at 2.0 mg/L above the potassium
permanganate demand for 2 h duration.
The survival of the group challenged and simultaneously treated with KMnO 4 (99%) was significantly higher than the positive
control (78%), challenged with F. columnare and not treated, and was not significantly different from the negative control
groups, not challenged. Fish treated with KMnO 4 post-challenge had 7% higher survival than the positive control (85% compared
to 78%) but that difference was not statistically significant. The results demonstrate that KMnO 4 has a clear prophylactic
value but probably a marginal therapeutic value once the infection has established.
BUYER PREFERENCES FOR SMOKED FRESHWATER FISH
Siddhartha Dasgupta*, Steven D. Mims and Jaime Greene
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
siddhartha.dasgupta@kysu.edu
Smoked fish is a value-added product that could open new markets to Kentucky’s aquaculture industry. This study was based
upon data from a USDA-funded marketing study of smoked freshwater fish. The relevant species used in this study were channel
catfish, hybrid striped bass, and paddlefish. Wholesale buyers, fish retailers, and restaurants were surveyed in Kentucky
and the Great Lakes area to evaluate their perceptions of the three types of smoked fish. Results showed a significant difference
in perceptions based upon the two geographic locations of the respondents. More Great Lakes respondents habitually purchase
smoked fish from individual suppliers, while all smoked fish buyers from Kentucky purchase from wholesale suppliers.
With respect to product ratings, smoked catfish compared poorly to the other products, while hybrid striped bass and paddlefish
were rated relatively highly. Most buyers considered the appearance of whole-smoked paddlefish to be superior to the other
fish types, and they felt that the paddlefish compared well with more traditional smoked fish such as sturgeon.

SELLING AQUACULTURE PRODUCTS IN KENTUCKY’S FARMER’S MARKETS:
RESULTS FROM MARKETING RESEARCH
Siddhartha Dasgupta*, Angela Caporelli and Kelly R. Probst
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
Siddhartha.dasgupta@kysu.edu
Kentucky has 111 farmers’ markets which are excellent outlets for locally-produced agricultural products. A survey of Kentucky’s
farmers’ markets had shown that almost 80% allow different meat and fish products to be sold. Kentucky State University,
in collaboration with the Kentucky Department of Agriculture, investigated the potential of selling different meat and
aquaculture products in Kentucky’s farmers’ markets. In this study channel catfish, tilapia, and freshwater prawns were evaluated
by farmers’ market customers.
Results showed the percentage of respondents who were familiar with aquaculture products in farmers’ markets (Table 1). Other
results showed that 52% and 50% of respondents were willing to buy prawn tails and whole prawns, respectively. However,
48% and 22% of respondents were willing to buy catfish fillets and whole catfish, respectively. Price was an important attribute
for 29% and 35% of respondents for tilapia and prawns, respectively. Similarly, 26% and 31% of respondents considered having
locally-grown tilapia and prawns, respectively, to be important.
Respondents picked different product attribute levels to construct and “ideal” tilapia product. The results show that most respondents
considered “ready-to-cook tilapia fillets, packed in 2 lb sections, priced at $6/lb or more” as ideal. With respect to
prawns, respondents considered “marinated whole animals in 2-lb packages, priced from $8-$10/lb” as ideal.
Table 1. Percentage of respondents who have seen the following products sold in Kentucky’s farmers’ markets.
Product Farmers’ market a
Ash Cor Lex Lou Fra Law Pad Erl Own
Catfish 0 33 42 15 15 20 26 0 4
Freshwater prawn 0 25 33 8 22 20 39 0 9
Number of observations 20 12 12 26 27 5 31 10 23
a Identification code for farmers’ markets (FM): Ash (Ashland FM), Cor (Corbin FM), Lex (Lexington FM), Fra
(Frankfort FM), Law (Lawrenceburg FM), Pad (Paducah FM), Erl (Erlanger FM), Own (Owensboro FM).
1

2
THE EFFECTS OF AQUACULTURE PRODUCTION NOISE ON THE GROWTH,
FEED CONVERSION, CONDITION FACTOR AND, SURVIVAL OF RAINBOW TROUT
Oncorhynchus mykiss
John Davidson*, Julie Bebak and Patricia Mazik
The Conservation Fund’s Freshwater Institute
1098 Turner Road
Shepherdstown, WV 25443 USA
j.davidson@freshwaterinstitute.org
Intensive aquaculture systems, particularly recycle systems, often require noise producing equipment such as pumps, blowers,
and filters that increase sound levels within the culture environment. These systems can be very effective at culturing healthy,
fast growing fish. However, the potential impacts of underwater noise on fish within intensive aquaculture systems have not
been evaluated. Field and laboratory studies have shown that fish behavior and physiology can be negatively impacted by noise.
Therefore, chronic exposure to aquaculture production noise could cause increased stress, reduced growth rates, and decreased
survival.
To further investigate, a study was conducted to evaluate the effects of aquaculture production noise on the growth, condition
factor, feed conversion efficiency, and survival of rainbow trout, Oncorhynchus mykiss. The study complemented previous
research, Wysocki et al. (2007), which evaluated the effects of production noise (115, 130, and 150 dB re 1 µPa RMS) on
rainbow trout. The present study evaluated the effects of the 115 and 150 dB treatments with increased tank replication, which
provided more statistical power, and analyzed additional growth parameters, including growth rates of individually PIT tagged
fish. These treatments represented sound levels lower than those recorded in an intensive recycle system and sound levels near
the upper limits known to occur in recycle systems.
Rainbow trout were cultured in experimental tanks (1.4 m 3 ) within a flow through system. To begin, mean fish weights in the
115 and 150 dB tanks were 40 and 39 g, respectively. After five months of noise exposure no significant differences existed
between treatments for growth, condition factor, feed conversion, or survival. Mean final weights for the 115 and 150 dB treatments
were 641 ± 3 and 631 ± 10 g, respectively. However, analysis of growth rates of individually tagged fish indicated that
trout from the 150 dB tanks grew significantly slower over the first month of noise exposure.
This study further suggests that rainbow trout growth and survival are unlikely to be affected over the long term by continuous
noise levels common to intensive aquaculture systems, but indicates that rainbow trout growth could be briefly inhibited by
sudden increases in noise. These results should not be generalized to all species since various species of fish have different hearing
abilities. Hearing specialist species such as catfish, carp, and minnows could be affected much differently by noise levels
common to intensive aquaculture systems.

WATER QUALITY AND TREATMENT EFFICIENCY IN REPLICATED RECIRCULATING
SYSTEMS OPERATED WITH LOW FLUSHING CONDITIONS WITH AND WITHOUT
OZONE
John Davidson*, Christopher Good, Carla Welsh and Steven Summerfelt
The Conservation Fund’s Freshwater Institute
1098 Turner Road
Shepherdstown, WV 25443 USA
j.davidson@freshwaterinstitute.org
Anecdotal evidence from previous un-replicated experiments at the Freshwater Institute indicates that rainbow trout mortality
increases and health declines with low water exchange rates and feed loading rates of 1.3-2.0 kg/d per m 3 /d of make-up water
flow. The decline in fish health appeared to be ameliorated by ozonation and was unrelated to an infectious disease. Typical
water quality parameters were also within safe limits. To further investigate, a series of controlled studies are being conducted
in six replicated recirculating systems (RAS) using rainbow trout.
This study compared rainbow trout performance and water quality criteria between three RAS operated with ozone and three
RAS without ozone. All systems were operated at low water exchange rates. Rainbow trout were randomly stocked, 1000
fish/system. To begin the study ozone was turned on and low water exchange rates (0.26 % of the total recycled flow) were
established. At this time fish in the ozone and no ozone RAS were 294 ± 1 and 296 ± 2 g, respectively. Each system recycled
380 L/min of water, which allowed a 15 min tank turnover rate. Each system contained a fluidized-sand biofilter, CO 2 stripper,
low head oxygenator (LHO), LHO sump, single 5.3 m 3 culture tank, drum filter, particle trap, and a pump sump. A 24-hr photoperiod
was provided. During a one week period when fish were at maximum feed levels (6.6 kg/day/tank) and densities (80
kg/m 3 ), water samples were collected across all unit processes to compare water quality and unit process removal efficiencies.
Mean feed loading rates for all systems were approximately 4.66 kg/d per m 3 make-up water flow. All tanks were fed equal
portions during the first month of the study and during the water sampling event. Thereafter, feeding rates were adjusted based
on satiation and waste feed observations.
Ozone addition resulted in substantially lower BOD, TSS, color, total particles, heterotrophic bacteria, higher UV transmittance,
and also appears to have removed dissolved copper (Table 1). Removal efficiencies across all unit processes were also
better for the ozone treatment. Statistical analysis for water quality is pending and will be presented. Ozone provided conditions
for improved growth. After five months, rainbow trout in the ozone and no ozone RAS were 1161 ± 6 and 993 ± 12 g, respectively
(p = 0.001). Cumulative mortality for ozone and no ozone was 7 ± 2 vs. 17 ± 5 morts, respectively (p = 0.145). Survival
was not statistically different but was better for the ozone treatment.
3

4
GREENHOUSE GAS EMISSIONS ASSOCIATED WITH DIRECT ENERGY INPUTS FOR A
WARMWATER LOW-SALINITY RECIRCULATING AQUACULTURE SYSTEMS
Max Davis*, Timothy J. Pfeiffer and Paul Wills
Florida Atlantic University
Boca Raton, FL. 33431 USA
bdavis35@FAU.edu
Recirculating aquaculture systems offer a solution to declining natural finfish stocks while limiting nutrient outputs and eutrophication
of effluent waters. Because water is reused in these systems, the solids must be efficiently removed by mechanical
treatment devices otherwise the solids buildup in these systems cause the cultured fish to be unpalatable and more importantly
unmarketable. This study evaluated particle size distribution and removal efficiency (RE) of three paired tank clarifiers in
three 43 m³ systems used in rearing juvenile red drum for stock enhancement purposes. The three clarifiers being tested for
removal efficiency were the 0.6 m³ swirl separator, 0.6 m³ static bed filter and the 0.6 m³ moving bed torrus filter. Water samples
were collected from the influent and effluent lines of each filter and serial fractionated through sieves ranging from 200 to 55
microns. Total suspended solid analysis was conducted on each set of influent and effluent samples to determine the particle
size distribution and percent removal efficiency of each filter type. The static bed filter had the highest removal efficiency for
total suspended solid particles (44% RE), and greater than 45% removal efficiency of all particles in the 200 through 55 micron
size range. The swirl separator had removal efficiency of over 45% for particles greater than 200 um. The moving bead torrus
filter had the least removal efficiency of particles between 200 to 55 microns but removed 9% of particles less than 55 microns.
Over 90% of solids in the influent and effluent samples were less than 55 microns in size the static bed demonstrated the best
removal of particles in this size range. The static bed clarifier removed 23% of particles in the less than 55 micron size range.
Overall, the static bed had the best removal efficiency of particles of all size ranges and was the most efficient clarifier. The
moving bed torrus filter had the second best total removal efficiency of 16% of which half of those particles were less than 55
microns in size.

ALTERNATIVE FEEDING STRATEGIES TO IMPROVE MILKFISH PRODUCTION
Evelyn Grace T. de Jesus-Ayson* and Russell J. Borski
Aquaculture Department
Southeast Asian Fisheries Development Center
Tigbauan, Iloilo, Philippines
edjayson@seafdec.org.ph
In the Philippines, cage culture of milkfish in marine environments is increasing. The practice uses high stocking densities,
with significantly greater inputs of artificial feeds which more often than not, has led to excessive feeding and consequently
excessive nutrient loading in receiving waters, exacerbating problems with pollution. These could have contributed to occurrence
of periodic fish kills in areas of marine milkfish culture clusters.
Sixty percent of milkfish farming expenses are attributable to feed costs. A series of experiments were conducted in an attempt
to develop alternative feeding strategies that will reduce feed inputs without compromising growth and resulting production.
In the first experiment, growth was compared in milkfish fed daily, on alternate days and on alternate 2-week or 4-week starvation
and refeeding cycles in a tank environment provided with flow-through water system. Results show, that milkfish fed on
alternate days do not grow as well as milkfish that are fed daily. Feed restriction for 2 weeks followed by 2 weeks of refeeding
elicited a compensatory growth response such that average body weight (ABW) of fish was not significantly different from
ABW of fish fed daily. Another experiment compared growth of fish given a ration equivalent to 10% of BW (usual practice)
or 7.5% of BW. Results show no significant effect on growth or final ABW or biomass. Thus, a lower feeding ration can be
given to milkfish without compromising yield. Results of the tank experiments will be tested in a simulated cage environment
in order to refine the feeding strategy and will be verified in actual marine cage and brackishwater pond production systems.

OVERVIEW OF PROPAGATION AND CULTURE OF PADDLEFISH BY STATE AND
FEDERAL HATCHERIES
Jan C. Dean*, Bobby C. Reed and Kristi C. Butler
Natchitoches National Fish Hatchery
615 South Drive
Natchitoches, LA 71457 USA
jan_dean@fws.gov
Spawning of wild paddlefish (Polyodon spathula) was observed and documented from the Osage River, Missouri, USA in
the spring of 1960. The first propagation of paddlefish occurred the following year with wild broodstock from the same area.
Further studies were done in the following years, and the first large-scale production of paddlefish was made at Blind Pony
State Fish Hatchery in Missouri in the early 1970s. Back then, culturists induced spawning using pituitary glands taken from
angler-caught paddlefish. The switch to induction via LHRHa took place in the early to mid 1980s based upon its success with
sturgeon spawning.
A survey was made of USA state and federal fish hatcheries regarding their techniques for propagation and culture of paddlefish.
State hatcheries produced paddlefish for sportfishing, recovery and educational purposes. Federal hatcheries did the same
for conservation, restoration and recovery. Broodstock sources included wild fish most often and also included hatchery reared
fish from the wild in areas of no prior paddlefish; two hatcheries received fertilized eggs from other sources. Spawning induction
was via LHRHa injections, with various rates for males. Egg collection included traditional stripping, Caesarian section
and the MIST procedure. Sperm collection generally was via a syringe with tubing. Most hatcheries used the semi-dry fertilization
method, whereas one used the dry method. Fullers Earth was used for egg deadhesion, and embryos were incubated
in McDonald jars primarily at 17-19 °C. Incubation water sources included wells most commonly though tap water and water
from a river and a creek was used. For the survey, production was divided into Phase I fish less than 7.5 cm length and Phase II
fish greater than this size. Phase I paddlefish were most often reared in raceways and less so in other tanks. They were offered
zooplankton and prepared diets. Phase II fish were reared in raceways and ponds primarily with some reared in circular tanks.
They were fed prepared diets most often. Harvest size ranged from 20 to 50 cm.
More specific information will be given for culture conditions at some hatcheries including Natchitoches National Fish Hatchery
and Booker-Fowler State Fish Hatchery, both of Louisiana. Stocking densities and growth rates will be included.

EVALUATION OF SOLVENT EXTRACTED, DEHULLED SOYBEAN MEAL FED IN
PRACTICAL DIETS TO LARGEMOUTH BASS Micropterus salmoides
William DeBoer*, Jeffery Weber, Bruce Cooper and Paul Brown
Department of Forestry and Natural Resources
Purdue University
West Lafayette, IN 47907 USA
wdeboer@purdue.edu
The purpose of this study was to evaluate the effects of graded levels of soybean meal in practical diets fed to largemouth bass
Micropterus salmoides. Solvent extracted, dehulled soybean meal was substituted into the diets for an isonitrogenous amount
of fish meal. Soybean meal concentrations were in increments of 10% of the diet and ranged from 0 to 60 %. Ten fish were
stocked into each of 21 tanks and held at 25 o C. Triplicate groups were randomly assigned one of seven different dietary treatments.
Each tank was fed to satiation twice daily for 8 weaeks. At the end of the study, fish were euthanized, and serum,
muscle, liver, and intestines from three fish per tank were collected and immediately flash frozen in liquid nitrogen and stored
in a -80 o C freezer. The corresponding tissue and serum samples were then used for metabolomic profiling. Additional intestine
samples were preserved in 10% neutral buffered formalin for histological examination.
Mean consumption of diet 1 (0% SBM) was significantly higher than diet 4 (30% SBM). There were no other significant differences
among consumption values. Weight gain of fish fed diets 2 and 3 (10 and 20% SBM respectively) were significantly
higher than in fish fed diet 4 (30 % SBM), but there were no other significant difference among treatments. Hepatosomatic
index of fish fed diet 2 (10% SBM) was significantly higher than fish fed diet 4 (30% SBM) and diet 5 (40% SBM). Feed
conversion ratio, specific growth rate, and visceral fat index were not statistically significant among treatments. Results of the
data indicate that largemouth bass can tolerate high levels of soybean meal in practical diets.

ANALYSIS OF CATFISH SUPPLY, DEMAND AND TRADE IN USA: BASELINE MODEL,
ESTIMATION STRATEGY AND PRELIMINARY RESULTS
Madan Mohan Dey*, Kehar Singh, Carole Engle and Abed Rabbani
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
1200 N University Drive, Mail slot 4912
Pine Bluff, AR 71601 USA
mdey@uaex.edu
The USA is the world’s largest producer of Channel catfish. The US catfish industry was one of the fastest growing businesses
in the 1980s and 1990s. But it has been on a contracting course over the last few years, and its production, imports and prices
scenarios are changing rapidly. The industry is facing higher feed price and strong competition from importers of catfish and
similar products like tilapia. Decision makers at different levels are facing with a number of what-if questions. What will
happen to the domestic prices of catfish if the production of catfish and competing products in the importing countries will
change? What if import unit value will change due to policy interventions? What if cost of production in the importing country
change? Quantitative modeling of catfish supply, demand and trade in the USA is a useful tool for analyzing recent structural
changes in the US catfish industry and for comparative analysis of policy options. The Aquaculture and Fisheries Center of the
University of Arkansas at Pine Bluff has recently constructed a model for the US catfish industry to analyze these issues. This
paper presents the structural version of the model, outlines the estimation strategy for specifying its parameters, and reports
preliminary results.
The model is composed of a production, consumption and trade core, and is specified to accommodate special features of the
US catfish industry. It integrates common features of partial equilibrium and gravity models. The model allows for changes
in tariffs and other policy instruments to be shocked to determine the resulting domestic price and quantity levels. It enables us
to assess effects on prices and quantities demanded and supplied under a variety of alternative policy options. The model also
provides us with forecast for catfish market prices, domestic demand and supply, and import demand and supply. To check the
accuracy of the forecast, the model uses Mean Absolute Forecast Errors worked out at different points of forecast.

WILL DECREASE IN POND BANK PRICES INCREASE THE CATFISH FARMERS’
INCOME IN U.S.? AN ANALYSIS OF PROCESSORS’ DEMAND BEHAVIOR
Madan Mohan Dey*, Kehar Singh, Carole Engle and Abed Rabbani
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
1200 N University Drive, Mail slot 4912
Pine Bluff, AR 71601 USA
mdey@uaex.edu
Pond bank prices of channel catfish have fluctuated substantially over the last two decades and there is a great deal of concern
related to how far prices may fall in future. There are many rumors and many speculations. Among others, there are comments
being made that pond bank prices must come down to increase the volume of catfish processed by processing companies. The
specific objectives of this paper are to : a) study the relationship between pond bank prices and processors’ prices, and pond
bank price and processing volume, b) analyze the nature of processors’ demand for catfish and the factors that affect it, and c)
analyze the impact of alternate pond bank and processors’ prices on fish farmers’ and processors’ income. The paper uses various
graphical, descriptive and econometric analyses. The analysis of processors ’ demand is based on a ‘partial adjustment
model’ assuming that the processors cannot totally adjust their demand to the desired level due to various constraints and rigidity
in the market. The model, estimated following Cochrane-Orcutt AR(1) regression, has explained 96.20 per cent variation
in the processors’ demand.
Analysis shows that there is a significant decline in average nominal pond bank price (by 4.3%) during the period from January
2000 to April 2008 as compared to the period before January 2000. However, the processors’ sale price (nominal) has decreased
only marginally over the period. The volatility in pond bank price has increased after 2000, but processors’ price has remained
more stable. The increase in the Consumer Price Index has been drastically higher than that of catfish prices (pond bank prices
and processors’ sale prices), indicating that the real price of catfish has decreased over the last decade or so. The estimated price
transmission function shows that the farmers receive about 94% of the processors’ sale price. The ratio has remained more or
less unchanged over the period, indicating that processors are not extracting a greater share from the farmers over time.
Econometric results reveal that the pond bank price, imports of catfish, basa & tra, and fuel price have significant negative
influences on the processors’ demand for catfish. The price elasticity of processors’ demand is highly inelastic, i.e., if there is a
1% decrease in pond bank prices, other things remaining the same, the processors ’ quantity demanded will increase, on average,
by 0.07 and 0.42% in the short run and long run, respectively. Decreases in pond bank price will cause further decreases
in farmer’s revenue, but processor will benefit marginally in terms of revenue gain. A 5% decrease in pond bank price (other
things remaining constant) will cause, on average, 3% revenue loss to farmers and 2% revenue gain to processors.

0
CHARACTERIZATION OF THE TAURA SYNDROME VIRUS ISOLATE ORIGINATING
FROM THE 2004 TEXAS EPIZOOTIC IN CULTURED SHRIMP
Arun K. Dhar, Dilip K. Lakshman, Keenan Amundsen, Refugio Robles-Sikisaka, Krista N. Kaizer, Sribash Roy,
Kenneth W. Hasson and F. C. Thomas Allnutt
Advanced BioNutrition Corporation
7155-H Columbia Gateway Drive
Columbia, Maryland 21046 USA
ADhar@abn-corp.com
Taura syndrome virus (TSV) is a major viral pathogen of penaeid shrimp worldwide. A comprehensive investigation of the
Texas isolate of TSV that caused epizootics in shrimp farms in Texas in 2004 (Us04Pv1) revealed that the virus was highly virulent
in laboratory bioassays causing severe symptom development and rapid mortality. Histopathology of moribund animals
demonstrated epithelial necrosis within the stomach, appendages, general body cuticle and gills. Animals that survived lethal
infection demonstrated moderate to numerous lymphoid organ spheroids (LOS), suggestive of chronic phase TSV infection.
In situ hybridization with a TSV-specific probe detected the virus in the LOS. Transmission electron microscopy of the virus
purified from infected shrimp showed that the virions are 31 nm in diameter with icosahedral symmetry. The genome of the
Us04Pv1 isolate was cloned, sequenced in its entirety and compared to five other geographically distinct TSV isolates for which
complete genome sequences are available in the GenBank database. Comparative genome analysis at the nucleotide and amino
acid levels showed that the Us04Pv1 isolate is more closely related to TSV isolates from Thailand and China and only distantly
related to the Venezuelan TSV isolate. Computational prediction indicated that the tertiary structures of the IAP and protease
domains of all six TSV isolates were very similar. However, the data from our 3-D modeling showed that the RdRp of Us94Pv1
has significant structural differences from the respective domains of five other TSV isolates. It remains to be determined if the
structural differences of TSV RdRps play a significant role in virulence or ecological adaptability of this isolate.

OUTREACH, ACCEPTANCE, AND SUCCESS OF POND AQUACULTURE IN PROMOTING
RURAL ECONOMY AND SOCIAL STABILITY
James S. Diana
School of Natural Resources and Environment
University of Michigan
Ann Arbor, MI 48109-1041 USA
jimd@umich.edu
Aquaculture has importance in promoting a rural economy and social stability in developing nations. The great majority of
the world’s aquaculture production comes from developing nations in Asia, where seafood production is important to both the
economy and local consumption. While export of seafood generated $42 billion for developing countries in 2006, still 75%
of all harvested seafood products were consumed in country. Aquaculture, which produced nearly 50% of the seafood harvest
for human consumption, is also important in both economic development as well as local consumption. The purpose of this
presentation is to review impacts of three aquaculture systems on social stability and rural economy in Thailand.
The first case, for prawn farming in Thailand, is focused on a local production system used to produce income. Prawn farming
in Thailand is mainly done to supply a local market; virtually none of the prawns are exported. Most farmer knowledge is
transmitted from farmer to farmer, with limited intervention of government or NGO extension agents. Prawn farmers generally
have small farms (averaging 6 ha), yet produce an income of about US $25,000 per year. They use intensive production with
aeration, frequent water exchange, and prepared feeds. While they have some problems in culture, 94% prefer prawn farming
to their former employment and all would like to make prawn farming more sustainable.
The second case is on tilapia farming in northeast Thailand as adoption of technology for increased food in the home. In this
case, considerable extension has been done to promote green pond (HiG) systems. Once again, farms were generally small (5
ha), with mixed farming of tilapia and other crops. Farmers mainly used tilapia to supplement their household consumption,
while some also sold surplus fish. People trained in HiG culture, or people who had been included in on-farm trials of HiG,
produced higher yields than those who learned of farming techniques from their neighbors. However, all groups valued their
fish farming activities and its contribution to their diet.
The final case focuses on shrimp farming and abandonment of shrimp ponds. Shrimp farmers mainly learn their techniques
from extension provided by CP, a feed company. Most farmers have a few ponds, lease their ponds, and grow shrimp for
export. Pond abandonment, due to disease or market failure, has been a big concern. However, only 14% of ponds had been
abandoned, while most were still in use for shrimp, polyculture, fish culture, or had been converted to housing. Farmers and
village leaders both agreed that shrimp farming has helped the local economy and provided jobs that are preferred over alternatives
of working in factories or salt farms, and they would like to see expansion of local jobs to keep families in their rural areas.
Shrimp farming has had a significant impact in the quality of life for rural farmers.
1

2
PHYSIOLOGICAL EVALUATION OF A CANDIDATE MARINE BAITFISH Fundulus seminolis
FOLLOWING SEAWATER ACCLIMATION
Matthew A. DiMaggio*, Cortney L. Ohs and B. Denise Petty
University of Florida
School of Forest Resources and Conservation
Fisheries and Aquatic Sciences Program
Indian River Research and Education Center
2199 South Rock Rd.
Fort Pierce, FL 34945 USA
mdimaggi@ufl.edu
US baitfish production had a 2005 farm gate value of $38 million. Aquaculture of marine baitfish species is still in its relative
infancy and increasing coastal property values are forcing marine aquaculture inland. Osmoregulatory limitations of candidate
marine aquaculture species need to be evaluated to determine feasibility of production and marketing. The seminole killifish,
Fundulus seminolis, is a naturally stenohaline freshwater killifish which has recently emerged as a candidate for marine baitfish
aquaculture.
Research specimens were collected from a local water body and screened for pathogens, preceding introduction into the research
system. Treatment groups were subjected to a gradual salinity change from 0 to 32 g/L over predetermined time periods
with freshwater controls for each corresponding time period. Acclimation times of 24, 48, 72, and 96 h were chosen with approximate
salinity increases of 5.3, 2.7, 1.7, and 1.3 g/L, respectively, every 4 h. The final salinity of 32 g/L was attained 4 h
prior to the termination of each treatment. Once the specified acclimation time had been reached the treatment group and its
corresponding control were sacrificed and blood and tissues were collected. Plasma osmolality, sodium, potassium, chloride,
hematocrit, change in body weight, and muscle water content were subsequently determined. Temperature, dissolved oxygen,
pH, TAN, nitrite, total alkalinity, and total hardness were monitored throughout the experiment.
Plasma sodium, chloride, and osmolality values were all significantly (p < 0.0001) elevated for all seawater acclimated fish
when compared with their corresponding freshwater controls. Body weight change revealed variable weight loss across all
control and saline acclimated groups; however, seawater acclimated body weight change values were significantly higher (p
< 0.0001) when compared with values obtained from control fish. Muscle water content percentages corroborated the body
weight change data, showing significantly less water (p < 0.0001) in the muscle samples of seawater acclimated fish compared
with their freshwater counterparts. The hypertonic seawater exposure significantly (p = 0.0007) elevated plasma potassium
concentrations in all but the 48 h acclimation treatment (p = 0.1000). Conversely, hematocrit percentages were variable and
only the 96 h acclimation treatment differed significantly (p = 0.0337) from the 96 h control.

EVALUATION OF A POINT OF CARE BLOOD ANALYZER FOR USE IN DETERMINATION
OF SELECT HEMATOLOGICAL INDICES IN Fundulus seminolis
Matthew A. DiMaggio*, Cortney L. Ohs and B. Denise Petty
University of Florida
School of Forest Resources and Conservation
Fisheries and Aquatic Sciences Program
Indian River Research and Education Center
2199 South Rock Rd.
Fort Pierce, FL 34945 USA
mdimaggi@ufl.edu
Conventional methods of hematocrit and electrolyte determination are gradually being replaced by new technologies. Point-ofcare
blood analyzers are both efficient and user friendly. As the use of such technologies becomes more pervasive in current literature,
investigations into the accuracy and reliability of point-of-care analyzers for evaluating hematological indices of fish is
warranted. There is currently little research validating point-of-care analyzers against conventionally accepted instrumentation
for use in fish. The purpose of this study was to evaluate a point-of-care blood analyzer (i-STAT®), and chosen cartridge (E3+)
(Heska Corp., Fort Collins, CO, USA) against conventionally accepted instrumentation for use in determination of hematocrit,
sodium, potassium, and chloride values in Fundulus seminolis.
The E3+ cartridge is designed to measure sodium, potassium, chloride, hematocrit, and hemoglobin from whole blood aliquots.
Hemoglobin measurements were not examined in this study because hemoglobin is not directly measured by the i-STAT®.
The E3+ cartridge was chosen because it is one of the most basic cartridges produced by the manufacturer yet its analyzed
parameters have application across numerous disciplines. Whole blood was used to measure parameters of interest in an initial
experiment. A second experiment implemented a whole blood heparin dilution prior to analysis with the point-of-care unit.
A final experiment examined the use of undiluted plasma in the point-of-care unit to determine electrolyte concentrations of
interest. The Bland-Altman method for assessing agreement between two methods of clinical measurement was used (Bland
and Altman, 1986) as well as calculations of correlation coefficients. A two tailed paired student t-test was also used to compare
values generated by both methods of analysis.
Generally, mean values obtained from conventionally accepted instrumentation were higher when compared with the i-STAT®
using whole blood, heparin diluted blood, and plasma. Significant differences (p≤0.05) were observed between all mean parameter
values analyzed by t-tests. Bias and limits of agreement values varied considerably throughout the experiments. None
of the blood parameters analyzed by the i-STAT® in this experiment could be considered reliable. Methodological validation
must be considered paramount for the introduction of new technologies in research applications.
3

4
EVALUATION OF THE USE OF ULTRASOUND TO STAGE MATURE WHITE STURGEON
Acipenser transmontanus FOR CAVIAR PRODUCTION
Brian C. Donahower*, Steve DuMond, Leo Ray, Linda Lemmon, Gary Fornshell, Terry Patterson, Jodi Rockett,
Madison S. Powell and Wendy M. Sealey
Hagerman Fish Culture Experiment Station
University of Idaho
3059F National Fish Hatchery Road
Hagerman, ID 83332 USA
donahower@uidaho.edu
Currently, the only means to assess ripeness of white sturgeon females and properly time harvest is measurement of oocyte
polarization index (PI). This technique requires minor abdominal surgery, is time consuming, and not an effective tool for
handling a large number of fish. Hence, oocyte PI is generally used only once in the fall to predict the timing of harvest of females
later in the winter and spring. This often results in an improper timing of harvest, decreasing caviar yield and quality, and
increasing the rate of follicular atresia (resorption of eggs). New methods to stage sturgeon for caviar production are needed to
replace the use of oocyte PI. These methods should be non-invasive, if possible, and quick.
Ultrasound has been employed in the medical field and terrestrial animal agriculture for several decades to non-invasively assess
soft tissue within live animals. Ultrasound is defined as acoustic energy produced by a transducer at megahertz frequencies.
As the wave energy passes through tissues, energy is scattered, absorbed or reflected back to the transducer depending on the
properties of the tissue and the relative changes of those properties between tissues.
To assess the utilization of ultrasound, 54 adult female white sturgeon and 4 adult male white sturgeon were scanned along the
abdomen in the transverse and sagittal planes. Ultrasonic images were captured at multiple sections along the left and right abdominal
walls between the pelvic and pectoral fins. Using ultrasound for initial evaluation, roughly half of the female sturgeon
were identified as having a distinct, mature egg mass on one or both sides of the abdomen. These findings were confirmed by
biopsy. Additionally, egg size was measured using the ultrasonic measurement tool and following biopsy with mixed results.
Female sturgeon not displaying clear signs of mature eggs using ultrasound were proven to have immature or atritic eggs.
While the number of male sturgeon examined was substantially less than female sturgeon, the presence of testes was unambiguous,
though the occurrence of large fat deposits surrounding the gonads made the detection of testes more difficult compared to
detection of female sex tissue. Preliminary results indicate that use of ultrasound can be an effective and non-invasive method
of sexing mature sturgeon and ascertaining the maturity of ovarian tissue in adult female sturgeon, though further analysis and
examination is warranted.

REPLICATION IN FIELD AND LAB EXPERIMENTS
Sara Duke
USDA- ARS SPA
2881 F&B Road
College Station TS 77845 USA
Sara.duke@ars.usda.gov
It is typically the goal of applied biological research to apply the results of controlled studies to a broader context than only the
local conditions of an experimental location. The issue of independent experimental replication is often confused with littleto-no
true replication in both large scale studies and experiments with restricted resources. Sub-sampling of an experimental
unit that has received a treatment is inadequate to estimate the experimental variability and the issue of pseudo-replication often
results. In order to make statistical comparisons to determine a meaningful treatment effect adequate replication is necessary.
Without reasonable replication, probabilistic statements of interpretation are limited to the very specific conditions or must be
only described for the experiment at hand; being conservative with the presentation of results. With low replication levels the
confidence intervals for effect size may be very broad help researchers understand their data better. Designing experiments
with adequate local replication and conducted at multiple-location are necessary to increase the scope of inference of experimental
results. By focusing on specific hypotheses that correspond with the original objective of a study, issues of multiple
comparison techniques and statistical power can be resolved and improved.
FISH DISEASES ONLINE COURSE IN VIDEO FORMAT
Robert M. Durborow* and Boris Gomelsky
Aquaculture Division
Kentucky State University
Frankfort, Kentucky 40601 USA
robert.durborow@kysu.edu
Online courses provide subject matter that may not be available at a learning institution where the student is enrolled. The
online approach is also convenient for the student who has limited time and may not be able to attend class at a particular hour.
Basics of Fish Diseases is an online course that is offered by Kentucky State University for 3 credit hours to undergraduate and
graduate students.
The course is presented on Blackboard ® in a recorded video lecture series. These video lectures can be viewed as often as the
student desires and can be downloaded as Pod Casts that can be transported on devices such as a video iPod. Blackboard ® allows
reading assignments, photographs and video clips to be posted in the Course Documents section. The incorporation of
video into the course is especially beneficial to students. The students are able to view movie clips of pathogens and diseased
fish. Video footage of sick fish swimming in a characteristic pattern and video microscopy of pathogens are more valuable to
students compared to still photographs.

MANAGEMENT OF AQUATIC WEEDS IN PUBLIC WATERS OF ARKANSAS- SPECIES AND
CURRENT CONTROL TECHNIQUES
Patricia Eklund*, Carole Engle and Mark Oliver
Aquaculture and Fisheries Center
University of Arkansas at Pine Bluff
Pine Bluff, AR 71602 USA
peklund@uaex.edu
Arkansas has an abundance of lakes and streams that provide excellent fishing and recreational opportunities to the American
people. The “Natural State” has over 90,000 miles of streams and rivers, over 600,000 acres of lakes, numerous bayous and
creeks, and some of the world’s most renowned trout fishing locations. People travel from all over the country to compete
in fishing tournaments and participate in other water-based recreational activities. Two of the most important tourist lakes in
Arkansas, Lakes Ouachita and DeGray, provide $3.6 billion annually to the state’s tourism industry alone. Native and non-native
aquatic weeds, although providing desirable game fish habitat, can restrict access for boating, swimming, and fishing in
public water bodies.
In Arkansas, the lead state agencies involved with maintaining public
waters are the Arkansas Game and Fish Commission (AGFC) and
the U.S. Army Corps of Engineers (USACE). A survey of the AGFC
District Fisheries Biologists (DFB’s) and a personal interview of a wildlife
biologist for the USACE were conducted to identify 1) the most
common aquatic weed problems in the respective AGFC districts, and
2) current management techniques used to control excessive growth in
public waters. Mail surveys were distributed to each DFB and a follow
up telephone interview was completed three months after the survey was
returned.
Eight AGFC districts of ten in total actively manage aquatic weeds in
AGFC-owned water bodies and the USACE actively manages weeds
in Lakes Ouachita and DeGray. Half of the AGFC Districts spend
greater than 90% of weed management efforts on native plant species,
specifically American lotus (Nelumbo lutea) and coontail (Ceratophyllum
demersum) (Figure 1). The USACE spend all of their management
efforts on the non-native Hydrilla (Hydrilla verticilata). Diploid grass
carp (Ctenopharyngodon idella) are the most common biological control
agent used by the AGFC for managing coontail and hydrilla while alligatorweed
flea beetles (Agasicles hygrophila) and Pakistani flies (Hydrellia
pakistanae) are used to control alligatorweed and hydrilla. Roughly
80,000 diploid grass carp stocked by the AGFC in 2007 into AGFC-
managed water bodies.

TWICE-ANNUALLY SPAWNING RAINBOW TROUT FEMALES IN A CULTURED
POPULATION FROM SOUTHERN CHILE
Francisco Estay*, Nelson Colihueque and Cristián Araneda
Piscícola Huililco Ltda.
Pucón, IX Región, Chile
fjestay@ctrnet.cl
Rainbow trout is characterized for being an annual spawner species, however it has been described that eventually it can experience
a twice annually spawning, determined by a double anually reproductive cycle (DARC), without photoperiodic manipulation.
This study was developed in the rainbow trout breeding center “Ojos del Caburgua” owned by Piscícola Huililco Ltda.,
located in Pucón, Chile.
About 1000 females three years aged of an autumn strain (Whyteville) were used during its first maturation time during the
2005 season. 934 spawns were registered during the autumn, and 321 specimens of the same group (36.2%), had an additional
spawn, during the next spring (Fig. 1). The reproductive performance of the normal and the additional cycle, was comparatively
evaluated (Table 1).
The average weight of the spawning females, the total
fecundity and the relative fecundity, exhibited significant
differences in favor of the additional cycle. Nevertheless,
the eggs’s diameter, the fertility rate and the
eyed eggs survival rate, were significantly higher in
the normal cycle, although, the eggs and fry obtained
from additional cycle, exhibited an acceptable performance,
compatible with the productive use of eggs.
On the other hand, the average eyed eggs per female,
were significantly higher in the additional cycle.
The additional reproductive cycle length, (LCRa) that
is to say, the period time from autumn spawning date
to spring spawning date, showed a statistically significant
increase, while the spring spawning season progressed.
The first spring spawner females registered
a LCRa average of 149 days (14/10/05) and the last
in spawning showed a LCRa average of 217 days
(03/02/06). Eggs diameter also registered a statistically
significant increases while the spring spawning
season progressed.
Macroscopic and histological aspect of ovary development
stages of mono and double-cyclers are also
showed and comparatively analyzed. Our aim is to
continue the research of the DARC character for improving
both the biological knowledge that supports
this character, and it industrial potential application.
Table 1. Comparative reproductive performance between normal and
additional spawn
Normal autumn Additional spring
spawn
spawn
Average ± S D Average ± S D
(n = 965)
(n = 321)
Female weight (g) 1,153 ± 217 a
1,512 ±176 b
Fertilization rate (%) 94.5 ± 7.5 a
77.8 ± 24.9 b
Total fecundity 2,691 ± 600 a
3,680 ± 509 b
Relative fecundity 2,271 ± 420 a
2,461 ± 354 b
Eggs diameter (mm) 4.89 ± 0.16 a
4.58 ± 0.23 b
Eyed eggs per female 2,341 ± 616 a
2,608 ± 678 b
Eyed eggs survival rate (%) 84.73 ±11.80 a
Different letters means statistical significative differences (P < 0.05)
68.84 ± 15.95 b

A PRELIMINARY STUDY ON THE USE OF CONCENTRATED ALGAE AS A
REPLACEMENT FOR LIVE ALGAE IN MARINE FINFISH LARVICULTURE
Elizabeth A. Fairchild*, Nathan Rennels and W. Huntting Howell
Department of Biological Sciences
University of New Hampshire
Durham, New Hampshire 03824 USA
elizabeth.fairchild@unh.edu
The addition of live microalgae into larval fish culture water to enhance fish growth and survival is standard practice in the
aquaculture industry. Microalgae or “greenwater” provides an added nutritional supplement to larval fish, as well as to the cultured
zooplankton fed to these fish. In addition, the green color change in the culture water provides a contrast which may aide
these young, developing fish in locating their prey. However, growing and maintaining live cultures of microalgae is a costly
and time consuming process. With the advances in algae processing, using concentrated solutions of microalgae for greenwater
are now possible. If larval fish growth and survival are as high using these greenwater products as they are with normal live
microalgae, the ramifications for the aquaculture industry could be significant.
To determine if concentrated algae products promote better growth and survival in larval winter flounder Pseudopleuronectes
americanus than live algae, and thus would be a suitable replacement, a preliminary experiment was conducted to test three
greenwater treatments: 1) control (no microalgae in culture water), 2) Nannochloropsis Instant Algae (liquid concentrate) from
Reed Mariculture, Inc., and 3) live Nannochloropsis grown on-site. Growth and survival of fish in the different treatments were
calculated for three weeks which coincides with the normal duration of greenwater use. Fish were monitored for another four
weeks to determine if the type of greenwater treatment affected long-term growth, survival, and pigmentation development.
Fish in all greenwater treatments grew each week and there was a lot of similarity in these growth rates. However, after three
weeks, fish from one of the Instant Algae tanks were significantly longer (p

CAN CROWDING DENSITY AFFECT ON BODY COMPOSITIO OF PERSIAN STURGEON
LARVAE?
Bahram Falahatkar, Mahmoud Shakoorian, Anooshirvan Ebrahimzadeh and Fereshteh Zahedi
Fisheries Department,
University of Guilan, Faculty of Natural Resources
Sowmeh Sara
P.O. Box 1144
Guilan, Iran
falahatkar@guilan.ac.ir
A 2-week culture of Persian sturgeon (Acipenser persicus) larvae was conducted with different densities in circular tanks. The
initial densities were 3500, 4000, 4500, 5000, and 5500 larvae/m 2 . Start weight of fish was 46.7 ± 6.8 mg with 18.3 ± 1.4 mm
length. Fish were fed with Artemia nauplii and then Daphnia every 3 h. At the end of the experiment, there were significant
differences in final weight among treatments, whereas the highest (167 ± 11.8 mg) and the lowest weight (129.2 ± 4.8 mg)
were observed at the lowest and highest density, respectively (Fig. 1). According to number of mortality, no significant differences
were observed between various densities. Proximate analysis of whole body showed no significant difference concerning
to moisture, protein, fat and ash contents. The Results of this study showed that high density of fish during larval rearing of
Persian sturgeon might affect on growth and some abnormalities but not on proximate composition during this short period of
rearing.
Weight (mg)
200
180
160
140
120
100
80
60
40
20
0
1 2 3 4 5
Larval Treatments
Fig. 1. Final weight of Persian sturgeon larvae held on different
stocking density after 2 weeks.

100
SEX IDENTIFICATION OF ASP (Aspius aspius taneatus) USING LAPAROSCOPIC METHOD
Bahram Falahatkar and Mohammad H. Tolouei Gilani
Fisheries Department
University of Guilan, Faculty of Natural Resources
Sowmeh Sara
P.O. Box 1144
Guilan, Iran
falahatkar@guilan.ac.ir
Knowledge of the sex of individual fish is useful for application to aquaculture or conservation management. However, the
determination of the sex of individuals with minimal handling or disruption of their ongoing activities can be a challenge.
Unfortunately, there is no external characteristic for determination of sex in some fishes such as Asp. The present study describes
laparoscopic methods for determining sex with observations of the gonads in Asp broodstocks. Six fish with 1270-3020
g weight, 52.5-73.6 cm length and 2-5 years age selected then anesthetized by MS 222 for testing gender via laparoscope with
inserting of cystoscope sheath into the body cavity with a small (1 cm) incision in the left part of abdominal wall with Hasson
technique and watched gonads onto the monitor. 100% of fish was correctly identified sex with the minimum duration of operation.
This study showed that the use of laparoscopy as a minimally invasive technique in this kind of fish could be efficient
method for sex identification and maturity stage in artificial propagation and stocks rehabilitation both for researchers and
aquaculturists.
Fig. 1. Using laparoscopy for sex determination of Asp
broodstock

CHANGES OF RELATIVE EGG WEIGHT DURING INCUBATION PERIOD OF PERSIAN
STURGEON, Acipenser persicus
Samaneh Poursaeid and Bahram Falahatkar*
Fisheries Department
University of Guilan
Faculty of Natural Resources
Sowmeh Sara
P.O. Box 1144
Guilan, Iran
falahatkar@guilan.ac.ir
The Persian sturgeon, Acipenser persicus, is an anadromous species endemic to the southern water of the Caspian Sea. In recent
years, more studies have been conducted in regards to identification of egg necessity of the Persian sturgeon in incubation
period. After fertilization the chorion of the egg stiffen, that is known as hardening, which serves to protect the developing
embryo also has an important role in fertilization. The egg weight increase, chorion hardening, is a completed mechanism based
on osmotic gradient (Alderdice, 1988) in which the action of different hardening enzymes like transglutaminase (Zotin, 1958),
protease and peroxidase (See Chang-Rak & Ichiro, 1998) are responsible for hardening. The increase in relative egg weight
could be used for evaluation of egg viability, understanding of changes during over-ripening and estimation of the best time
for egg enrichment via some micronutrients solution. Therefore, the aim of present study is to determine egg quality, trends of
relative egg weight, and the peak time of water absorption in Persian sturgeon during incubation period.
Experimental eggs were obtained from 20 females of Persian sturgeon caught from southern parts of the Caspian Sea. Each
female was stripped and coelomic fluid was removed from eggs before insemination. The subsamples were weighed for determination
of the weight of non-water-hardened eggs. Eggs were fertilized with a mixture of sperm from 2 males, checked sperm
activities before fertilization. The average activity of sperm was 85.6 ± 7.3%. Because of stickiness of eggs after fertilization,
silt solution was used according to Kohnehshahri and Azari (1974). Fertilized eggs was rinsed with hatchery water and then
transferred to the Youshchenko incubators for incubation. The eggs were sampled 5 min (after fertilization time), 60 min (after
adhesive removing), 2, 3, 4, 5, 6, and 7 h after fertilization (during incubation). Finally, the number of eggs was counted for
each subsample (1-2g) to calculate the absolute egg weight. Data was analyzed with SPSS software using one-way ANOVA
and Tukey’s as post-hoc test. Data are presented as mean ± S.D.
The results of this study showed that the highest water uptake was observed during 3h after fertilization (AF), whereas no
significant differences was found until 7h AF (P>0.05). Egg weight increased significantly during fertilization and water absorbance
up to 60.6% compare to 3h AF (P

102
THE ANESTHETIC EFFECT OF TRICAINE METHANSULFONATE ON PERSIAN
STURGEON LARVAE (Acipenser persicus)
Bahram Falahatkar*, Nesa Heshmat Solati, Sobhan Ranaye Akhavan and Samaneh Poursaeid,
Fisheries Department
University of Guilan, Faculty of Natural Resources
Sowmeh Sara
P.O. Box 1144
Guilan, Iran
falahatkar@guilan.ac.ir
Anesthesia is generally defined as a state caused by an applied external agent resulting in a loss of sensation against external
and internal stimulants inhibition of reflex activity, and reduced skeletal muscle tone through depression of the nervous system.
Anesthesia plays an important role in both fisheries research and aquaculture, being used to reduce stress in fish for high
density, catching, transportation etc to delicate handling operation. However, inappropriate using of anesthetics could cause
to deleterious effects. Tricaine (3 aminobenzoic acid ethyl ester methanesulfonate or MS 222 ) has been shown to be useful for
anesthetizing larval fish. The objective of this study was to evaluate the best dose, anesthetic and recovery time in Persian
sturgeon larvae.
For determining the best dose of anesthetic, different concentrations of tricaine methanesulfonate including 0, 25, 50, 75, 100,
150, 200 mg/L with three replicates were compared on Persian sturgeon (Acipenser persicus) larvae. Thus, 10 larvas with mean
total length and body weight 3.63 ± 0.54 cm and 241.06 ± 7.01 mg, respectively, were put on 100 mL of MS 222 solution. Afterward,
recovery was done by means of putting of fish into 200 mL clean water. Dissolved oxygen and temperature were 7.11 ±
1.4 mg/L and 20.26 ± 0.05, respectively.
Based on the results of mortality rate, duration of anesthesia, recovery time, reaction to the anesthetic, it is recommended that
the concentration of 75 mg/L is the best dose for anesthesia in Persian sturgeon larvae. These results showed that MS 222 is an
effective anesthetic material for this unique species.

CONSUMER SAFETY ASSESSMENTS FOR CULTURED SPECIES AT UNBRANDED
MARKETS
José Fernández-Polanco, Ladislao Luna and José Luis Fernández Sánchez
Research Group in Economic Management for Sustainable Development of the Primary Sector
University of Cantabria
Av. de los Castros
s.n. 39005 Santander, Spain
polancoj@unican.es
lunal@unican.es
fernandezjl@unican.es
1. Introduction
Brands and other identifications are common extrinsic cues used by consumers to assess quality of purchased foods. Effects on
food purchase decisions have been proven in the seafood literature (Wessells & Anderson, 1995; Wessells et al., 1999). Safety
has been treated as an aspect of the perceived quality in food, so branding also contributes to assure it. This paper studies factors
affecting consumer safety assessments when there are no brands to use as cues, as happens in the Spanish markets with most
aquaculture species. Alternative sources of information are tested as causes of variations in the safety scores of three cultured
marine species.
2. Methodology
A perceived safety index is computed from the scores provided by cultured sea bream, sea bass and turbot consumers, extracted
from a sample of 3.200 Spanish seafood consumers conducted in November 2005. Lickert scales relating to consumer beliefs
about aquaculture, official generic advertising campaigns credibility and importance of shop and seller in the food choice decisions
were answered by a total of 220 respondents. Scales linked with usual consumer processes with branded foods were also
included in questionnaire. A path analysis model is performed using a distribution free algorithm for test causal effects over
safety index.
3. Results
Usual behaviour models for branded products fail to explain the
perceived safety of cultured species in the referred market. Brands
absence prevents the search for special product characteristics and
the willingness to pay a premium. Instead, a model considering
other extrinsic information sources satisfactory fits (Table 1).
Perceived safety of aquaculture products is positively affected by
the health benefits expected by consumer and confidence in the
fishmonger prestige. In the opposite, confidence in seller suggestions
adversely affects perceived safety. Last is higher in the traditional
seafood marketing channels.
The beliefs on potential benefits for health arising from
cultured species consumption are positively determined, in turn, by
general belief about the safety of aquaculture methods and the generic
advertising campaigns credibility.
4. Conclusions
Brands absence at Spanish seafood markets hampers the development of strategies seeking for a better product appraisal.
Greater importance of the distribution channel agents on the consumer safety perception derives from results. Consumers
will consider the cultured species more safety as greater the shop prestige. Seller suggestions favour other larger margins fish
sources, penalizing aquaculture.
103

104
LC50 DETERMINATION OF ENDOSULFAN FOR THE Rutilus frissi Kutum FRY IN IRAN
Firouzbakhsh, Farid
Fish Disease laboratory
Department of Fisheries
Sari University of Agricultural Science & Natural Resources
Mazandaran- Sari-Iran
f.firouzbakhsh@umz.ac.ir
Endosulfan is a chlorinated pesticide widely used in Iran. The LC50 of endosulfan for the Ruilus frissi kutum fry with mean
body weight 150 mg was determined. The study was conducted in Mazandaran province, north of Iran. The LC50 96h value of
endosulfan was 0.156 micro liters per liter. All of important physicochemical parameters of water were monitored and maintained
at a constant value so that temperature was 22 ± 1 degrees centigrade, PH was between 7.2 – 7.8 and dissolved oxygen
was around 6 mg/l. Three treatments and three replicates were run for each treatment. The software used was LC50 Model
System version 1.0 that let estimation of LC50 values by Spearman-Karber rule with (P≤ 0.05).

SEAFOOD AT ITS BEST: A FOUR-LESSON CURRICULUM ENCOURAGING EVERYONE
TO EAT MORE FISH
Gary Fornshell and Rhea Lanting
University of Idaho Extension
Twin Falls, Idaho 83301 USA
gafornsh@uidaho.edu
rhlantin@uidaho.edu
Current per capita consumption of all seafood is approximately 142 g raw [seafood] per week, only about 63% of the American
Heart Association recommendation of 227 g raw. Low seafood consumption can be explained in part by concerns and confusion
about seafood safety, handling, and cooking. Many of these concerns result from inaccurate and sensationalized media reports.
American consumers are also confused by conflicting information from governmental agencies and private organizations.
Several prominent organizations recommend increased consumption of seafood for the many important health benefits. These
include, the American Heart Association (at least two 113 g serving per week), the 2005 Dietary Guidelines Advisory Committee
(two 113 g servings of fatty fish per week), the American Diabetes Association (2-3 servings per week), and the World
Health Organization (1-2 servings per week equivalent to 200-500 mg of EPA+DHA).
Improving and maintaining health is one of the most important factors consumers cite when considering their reasons for eating
seafood. A complex informational environment influences food choice. Factors that influence choosing seafood are similar to
choices of other foods, such as taste, price, convenience, or ease of preparation. Consumers of seafood do not think they have
enough information about seafood. Those enrolled in this curriculum will increase their knowledge of the following:
• Where seafood comes from;
• Benefits and risks of eating seafood; and
• How to select, handle, store and prepare seafood.
Increased knowledge can lead to better informed consumers and increased consumption of seafood.
The target audience for this curriculum includes: family and consumer sciences educators, nutritionists, dieticians, food service
personnel, and nutrition outreach professionals. Seafood At Its Best provides an opportunity for teaching the 2005 Dietary
Guidelines for Americans recommendation for “choosing fats wisely for good health,” using seafood as an example of a food
source low in fat yet containing the good fats. The curriculum is designed to educate clientele about seafood, often poorly understood
in terms of selection, handling, storage, preparation, health benefits, and actual risks relative to perceived risks.
10

10
EFFECT OF DIETARY STEARIDONIC ACID-CONTAINING SOYBEAN OIL ON GROWTH
AND FATTY ACID COMPOSITION OF PACIFIC WHITE SHRIMP Litopenaeus vannamei
Ian P. Forster, Warren G. Dominy, Leonard Obaldo, Gary F. Hartnell, Martin C. Ruebelt, Tim C. Hickman
and Ernest Sanders
Oceanic Institute
41-202 Kalanianaole Hwy.
Waimanalo, Hawaii 96795 USA
iforster@oceanicinstitute.org
Typically, shrimp diets are supplemented with 5-9% fish oils containing n-3 fatty acids to ensure maximal growth and development.
A study was undertaken to ascertain the effect of feeding diets containing stearidonic acid (SDA)-containing soybean oil
on growth performance and sensory characteristics of shrimp L. vannamei. Six diets (35% crude protein, 8.4% ether extract)
were prepared with 3 levels of soybean meal (SBM) in replacement of menhaden fish meal (FM), with supplemental menhaden
fish oil (FO) or SDA modified soybean oil. Juvenile shrimp (1.7 g) from the breeding program at the Oceanic Institute were
stocked into 24 black fiberglass cylindrical tanks (100 shrimp per tank) filled with ca. 1,000 L of seawater. Flow-through water
was supplied at the rate of 4 L/min. Each diet was fed for 12 weeks to 4 tanks of shrimp to apparent satiation. Performance data
were analyzed by 2-way ANOVA with a 5% error rate. Fatty acid profiles of the whole body and tail muscle samples of shrimp
in each treatment were compared with those of the corresponding diet.
Shrimp fed the diets containing FM had the highest growth overall (1.58 g/week), with shrimp fed diet with the SDA oil performing
as well as those fed the FO diet. At increasing levels of SBM addition, however, shrimp fed the SDA oil exhibited
lower final weight and growth rate than those fed the corresponding FO diet. The feed conversion ratios (FCR) of shrimp fed
the SDA oil-containing diets were not different compared to the corresponding FO diets, except that FCR was lower in the 50%
FM replacement groups with SDA oil, relative to the FO group. Survival was not affected by treatment.
In general, the fatty acid (FA) composition of the shrimp muscle and whole body reflected the FA composition of the corresponding
diets. In the shrimp fed diets containing FO, the SDA was much lower than in the shrimp fed the diets with the SDAcontaining
soybean oil. Among the shrimp fed the FO, there was little variation in the range of whole body and tail muscle. In
contrast, in the shrimp fed the SDA-containing soybean oil, the mean whole body and tail muscle EPA and DHA were greatly
reduced as the soy oil was increased. The reduction in EPA and DHA in shrimp fed the soy oil containing diets, was much less
than the increase in SDA in the tail muscle and whole body of the shrimp. Levels of EPA and DHA in shrimp fed the 0 SBM
diet were similar whether the diet contained FO or the SDA containing soy oil.
It is concluded that SDA-containing oil can replace FO in diets for juvenile shrimp cultured in clear water with no reduced
growth, when there is sufficient FM in the diet. Under conditions of significant replacement of FM by SBM, replacing FO with
SDA results in reduced shrimp growth, although the effect on FCR is less clear. In this trial, SDA-containing soy oil is unable
to completely meet the needs of shrimp for n3HUFA suggesting that conversion of SDA to HUFA is limited in this species.

OPTIMIZATION OF SHRIMP DIETS USING MIXTURE MODEL METHODOLOGY
Ian P. Forster*, Warren G. Dominy, Zhi Yong Ju, Addison L. Lawrence and Susmita Patnaik
Oceanic Institute
41-202 Kalanianaole Hwy.
Waimanalo, Hawaii 96795 USA
iforster@oceanicinstitute.org
Meeting the nutritional needs of animals in culture conditions generally requires assembling a selection of ingredients. The
mission of the nutritionist is to develop diets that meet the needs of animals efficiently at lowest cost and with lowest level of
waste. Many strategies are used to accomplish this, including measuring the effect of several combinations of ingredients in
diets. By carefully selecting which combinations of ingredients to test in this way, it is possible to efficiently optimize diets
for specific purposes. Using a factorial approach to examine one or two ingredients at a time can be very resource intensive in
relation to the information generated. Design of experiments using mixture model methodology is an alternative method that
enables optimization of ingredient combinations using fewer resources than a factorial approach. Under mixture model design,
diets are produced in which the ingredients to be examined comprise a fixed level of all diets, with the remaining portion of
each diet held constant. Using this approach, it is possible to examine the ability of ingredient combinations to maximize desired
response (e.g., growth, feed efficiency), not only at fixed levels, but also at different nutrient class levels.
A series of trials was run using mixture model methodology to optimize diets for Pacific white shrimp Litopenaeus vannamei,
using krill meal, menhaden fishmeal, squid meal and soy protein isolate. Combinations of squid meal and soy protein isolate
were considered at different protein levels and lipid sources were considered at different lipid inclusion levels. An important
element of the research in this study was the use of base mixes which were balanced for known and presumed requirements for
protein (except in one trial), lipid (except in one trial), calcium and phosphorus. Balancing the base mixes resulted in all diets
being balanced. The diet trials were run with juvenile shrimp in flow through water, in indoor culture tanks for five week duration,
with 10 or 12 tanks (depending on trial) assigned to each diet. Final weight (g), growth (g/week) and survival (%) were
applied to the mixture model determine the interactive
It was found that optimum final weight and growth of the shrimp was achieved with a diet containing soy protein isolate 15.4%
and squid meal 30.8%, with a dietary crude protein of 40%. Both krill and fish meals were eliminated from this diet.
10

10
CAN DIETARY LINOLENIC ACID ALONE SUPPORT GROWTH OF THE FRESHWATER
CATFISH Rhamdia quelen?
Débora M. Fracalossi*, Rodrigo J. Vargas and Alexandre S. Garcia
Aquaculture Department
Federal University of Santa Catarina
Florianópolis, SC - Brazil
deboraf@cca.ufsc.br
Jundiá is a siluriform with wide geographical distribution and commercial importance in Southern Brazil. Linolenic acid (18:3
n-3, LNA) is an essential fatty acid for freshwater fish but its dietary requirement was not yet established for jundiá. Previous
studies showed that jundiá fingerlings fed different dietary lipids have a high capacity of elongation and desaturation and are
able to utilize dietary LNA to produce fatty acids with 20 and 22 carbons. Therefore, a study was carried out to evaluate the
effects of five dietary concentrations of LNA (0.01%, 0.10%, 0.23%, 0.36%, and 0.43% dry basis) on growth of jundiá fingerlings.
Such LNA concentrations were achieved by mixing different vegetable oils to an isonitrogenous (~40% crude protein)
and isoenergetic (~11.5 kJ/g) casein-based basal diet. These diets did not contain detectable amounts of 20:5 n-3 (EPA) nor
22:6 n-3 (DHA). Additionally, the basal diet supplemented with refined fish oil, was used as a control. Diets were randomly
assigned to 18 groups of jundiá fingerlings housed into 70-L tanks on a water recirculation system (29 + 1.2ºC) at a stocking
density of 0.6 g/L. Jundiá fingerlings, with an initial body weight of 1.5 g + 0.04, were fed twice a day to apparent satiation for
70 days.
Fish survival was superior to 94% for all diets. A positive correlation (R 2 =0.721, p=0.0005) was found between dietary LNA
concentration and weight gain (Figure 1). A similar correlation was found between dietary LNA concentration and specific
growth rate (SGR=3.16+0.65LNA+2.14LNA 2 , R 2 =0.66, p=0.0015). A comparison between the control and the experimental
diets showed that fish fed a mixture of vegetable oils providing 0.36 to 0.46 LNA (% diet) for 70 days had similar growth to
fish fed refined fish oil (Figure 1). These findings suggest that long-chain highly-unsaturated fatty acids are not required for
jundiá growth at high water temperature. Further studies for longer feeding periods and at lower water temperatures will help
to better understand the role of dietary LNA on growth of jundiá fingerlings.

EVALUATING POND AQUACULTURE EFFLUENTS THROUGH BIOLOGICAL
ASSESSMENT OF FISH AND BENTHIC MACROINVERTEBRATE ASSEMBLAGES IN
RECEIVING STREAMS
Emmanuel A. Frimpong* and Steve Amisah
Department of Fisheries and Wildlife Sciences (0321)
Virginia Polytechnic Institute and State University
156 Cheatham Hall
Blacksburg, VA 24061 USA
frimp@vt.edu
Aquaculture is a promising alternative to capture fisheries, especially in developing countries where most capture fisheries are
already over-exploited and the value of small streams and headwaters as prime protection for large, downstream productive
rivers is often threatened by unsustainable agricultural practices. Some of the most intimate relationships between farmers and
streams are found in the use of streams as a source of clean water for filling earthen ponds and as conduits of effluents from
ponds. As a supporting system for pond aquaculture, streams are vital to the sustainability of aquaculture industries, but are also
directly threatened by harmful effluents in the absence of best management practices.
Developing countries need to develop inexpensive methods for monitoring pond effluent impacts on receiving waters and adopt
management practices that maintain the quality of streams to support aquaculture development. This study gives an overview
of ongoing research work in Ghana in partnership with Virginia Polytechnic Institute and State University and the Aquaculture
and Fisheries Collaborative Research Support Project to develop ecologically-based effluent monitoring protocols for aquaculture.
A vital first step in the development of a biological monitoring protocol is standardization of sampling techniques to collect
representative samples of biota. While standardized stream fish and macroinvertebrate sampling is now routinely done is temperate
and developed regions (for example, in the United States), tropical streams present unique sampling challenges for biological
assessment and little is known about the best way to collect representative samples of tropical stream biota. We present
preliminary results of the performance of different configurations of seining and kick-sampling and benthic cores as standard
methods for fish and macroinvertebrate sampling, respectively, in tropical streams. Methods for developing assessment metrics
based on the two types of biological assemblages will also be discussed. Biological assessment and best management practices
provide a viable, low-cost balance to the growth of aquaculture in developing countries.
10

110
AN IMPLANTED MINITATURE ACOUSTIC TAG EVALUATION ON SUB-YEARLING FALL
CHINOOK SALMON Oncorhynchus tshawytscha
Deborah Frost*, R. Lynn McComas and B. Sandford
NOAA Fisheries
Northwest Fisheries Science Center
Manchester Research Station
7305 Beach Drive E.
Port Orchard, Washington 98366 USA
Debbie.Frost@noaa.gov
Implantable acoustic tags are often used to assess salmonid smolt behavior, survival, habitat use and predation risk. However,
the relatively large size of some acoustic tags limits their use in juvenile fish. A miniature implantable acoustic tag prototype
(21 mm by 7 mm by 6 mm and 0.9 g in air) was evaluated in three separate groups of Chinook salmon Oncorhynchus tshawytscha
smolts for impacts on 1) growth and survival, 2) predation risk and 3) behavioral differences. Growth rate over a 30-day
period was significantly affected by the tag (P = 0.003 for length and 0.000 for weight, Table 1). Tag to body weight ratio
ranged from 2.6 to 5.9% at tagging. Survival was not affected, however, partial to complete tag expulsion was noted in 37% of
the tagged fish in the growth evaluation group. Implantation of an active miniature prototype acoustic tag did not significantly
affect predation risk (P = 0.255). Non-significant differences in behavior involving activity level (P = 0.467) and position in
the water column (P = 0.298) (Figure 1) were also seen, however the small sample sizes in both the predation and behavior
evaluations suggested trends toward an affect by either the tag or the surgical procedure. These evaluations showed a tendency
for growth and tag retention to affect the overall successful use of this size of tag in this size of fish by impeding growth and
potentially contributing to false mortality with expulsed tags. The study emphasizes the need for careful evaluation for longterm
effects prior to field use.

EVALUATION OF STRESS-INDUCED CORTISOL RESPONSE FOR USE IN A Morone
SELECTIVE BREEDING PROGRAM
S. Adam Fuller* and Matthew McEntire
United States Department of Agriculture
Agricultural Research Service
Harry K. Dupree Stuttgart National Aquaculture Research Center
Stuttgart, AR 72160 USA
Adam.Fuller@ars.usda.gov
Harry K. Dupree Stuttgart National Aquaculture Research Center (HKDSNARC) has initiated a selective breeding program
aimed at improving production traits for sunshine bass, Morone chrysops X M. saxatilis, via multi-trait selection in the two
parental species, white bass, Morone chrysops (Rafinesque), and striped bass, Morone saxatilis (Walbaum). We are interested
in understanding the physiological and genetic impact of many production trait phenotypes that are currently being characterized
in white bass and striped bass for the hybrid striped bass National Breeding Program. Cortisol is a hormone steroid with
many biological effects, including gluconeogenesis and immunosupression. If selective breeding could be used to reduce the
severity of the physiological stress response it would be a benefit to those culturing hybrid striped bass.
A cortisol dose response curve was created using three
year-old white bass exposed to a low water stressor for
0, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, or 80 minutes.
At each time interval, a blood sample was taken from
each of five fish with a heparinized syringe using no
anesthetic, the blood centrifuged and the plasma stored
frozen. Plasma cortisol concentrations were determined
by radioimmunoassay (RIA) (Figure 1). Relative stress
response was then determined for three year-old white
bass broodstock representing 80 crosses produced at
the North Carolina State University’s Pamlico Aquaculture
Field Laboratory. Male (n = 49) and female
(n = 85) white bass were segregated, tagged, then
weighed, measured, and stocked into 700-L tanks at a
rate of ten same sex fish per tank. Fish were exposed to
a low-water stress event for 40-minutes then captured
and bled without anesthesia. This was done once per
month for two consecutive months. Cortisol concentrations
were determined as described.
There was a significant difference in mean cortisol/g
body weight response between the male (0.409 ± 0.020)
and female (0.224 ± 0.016) white bass (P < 0.0001)
(Figure 2). The lowest-responding 25% of females
(cortisol/g < 0.329) and males (cortisol/g < 0.548) will
be selectively bred to determine heritability of low cortisol
response.
111

112
ANALYSES OF VARIABLE NUMBERS OF TANDEM REPEATS IN ORF 94 OF WHITE
SPOT SYNDROME VIRUS (WSSV) ISOLATES FROM OUTBREAKS IN MEXICAN SHRIMP
FARMS
Lucio Galaviz-Silva*, José Reyes González-Galavíz, José C. Ibarra-Gámez and Zinnia J. Molina-Garza
Facultad de Ciencias Biológicas
Universidad Autónoma de Nuevo León
Nuevo León, México
lgs12167@yahoo.com
White spot syndrome virus (WSSV) causes the most serious losses to commercial shrimp farming worldwide. Since its discovery
in Fujian, China (1991) the WSSV spread quickly from South-East-Asia to United States in 1995; Central, South America
and Mexico since 1999; and in 2002 the virus was also reported in France and Iran. Sequence comparisons of genomic regions
have shown very little variation in widely separated geographical isolates of WSSV. Recent publications have revealed a variable
numbers of tandem repeats (VNTRs) in the region encoding ORF94. In all WSSV isolates from Thailand or Vietnam,
ORF94 has tandem repeat units (RUs) of 54 bp, with a single nucleotide polymorphisms at position 48 (G or T). This genetic
variation may be useful as a genetic marker for strain identification and epidemiological studies and could be correlated with
the spread of WSSV.
The origin of the collected WSSV-infected shrimps (Litopenaeus vannamei) was shrimp ponds experiencing outbreaks of
WSSV, located in Sinaloa (Tarahumara Duran) and Sonora (Gutierrez Acuicola, Acuaproductiva and Remanentes). Infected tissues
were homogenized in TN buffer and DNA was isolated with proteinase K and sarkosyl, followed by phenol-chlorophorm
extraction. The primers used were derived from ORF94 of WSSV (Wongteerasupaya et al. 2003. DAO 54:253-257). PCR products
were purified and cloned in the pGEM-T easy vector system (Promega). Plasmids were prepared with DNA purification
system (Promega) and sequenced using an automated ABI 3130 Sequenced system (Applied Biosystems). Sequence alignment
and phylogenetic analysis were conducted using Clustal W and GeneDoc software. Our results from 4 outbreaks ponds shrimp
indicated the evolutionary variability of the WSSV. Based on this test, we assumed that each pond outbreak was caused by 4
different isolates of WSSV. An example is showing in Table 1 for fragments representing 2, 4, 6 and 7 repeats. Six and seven
RUs of 54 bp have been reported in Thai isolates, but the presence of 2 and 4 RU´s could suggest two WSSV isolates natives
from Mexico, in accordance a phylogenetic analysis (Fig. 1).
Table 1. Number of ORF94 repeats found in DNA extracts of samples from 4 WSSV
outbreaks ponds.
Samples Amplicon
size (bp)
No. of
repeats
T. DURAN 235 2
ACUAP 398 4
REM 506 6
GEZ
ACUICOLA
560 7
Fig 1. Bootstrap analysis of unrooted
phylogenetic trees of an isolate of WSSV from
Sonora (508) was located in a separated cluster.

PERFORMANCE OF Litopenaeus vannamei (BOONE, 1931) CULTURED PROBIOTIC WHEN
SUBMIT TO INFECTION BY Vibrio harveyi
Danielli Dantas, Wanessa Costa, Marcelo Rego, Roberta Soares, Silvio Peixoto and Alfredo Gálvez *
Universidade Federal Rural de Pernambuco
Departamento de Pesca e Aqüicultura
R. Dom Manoel de Medeiros
s/n, Recife-PE, CEP 52171- 900, Brasil
alfredo_oliv@yahoo.com
The fast expansion of farmed shrimp industry during the last decades has been offering high profitable deal to many countries.
However, diseases appearing have caused lots of serious economic damage on productive sector. The effect of probiotic in
Litopenaeus vannamei juveniles when submitted to the infection by Vibrio harveyi was evaluated in the present study.
The exposure to V. harveyi was carried out during 15days using individuals reared for one month in a closed recirculation system
with a commercial probiotic (Bacillus sp.) The Probiotics were added to water and food (PWF), only in the water (PW),
only in food (PF) and without probiotic (WP).
The food consumption did not differ significantly among the treatments, but in the WP treatment a significant reduction occurred
in the second week (Table 1). The PWF and PW treatments showed inferior and more constants values in the total hemocits
concentrations during the experimental period. The highest growth rates were observed in the PWF (0.34 ± 0.19 g/week)
and SP (0.45 ± 0.04 g/week) treatments, but did not differ statistically within these treatments. Shrimp survival values were
higher than 98% in all treatments.
The results suggest that the addition of Bacillus sp. to the water or food, keeps the food consumption during a longer period of
time in shrimp infected by V. harveyi and interfere positively in the immune response of L. vannamei.
Table 1. Mean and standard deviation of the Litopenaues vannamei shrimp feed consume (gram of feed/food) in closed system with Vibrio harveyi
inoculation *
Period PWF PW PF WP
7º day 0,79 ± 0,27 a
14º day 0,84 ± 0,33 a
* Different letters indicates signicant different (p

114
OXYGEN CONSUPTION OF Litopenaeus vannamei JUVENILES IN A SUPER INTENSIVE
ZERO WATER EXCHANGE RACEWAY CULTURE
Luis Vinatea, Alfredo O. Gálvez*, Jesus Venero, John Leffler and Craig Browdy
Universidade Federal Rural de Pernambuco, Departamento de
Pesca e Aqüicultura, R. Dom Manoel de Medeiros
s/n, Recife-PE, CEP 52171- 900, Brasil
alfredo_oliv@yahoo.com
It was studied the rate of dissolved oxygen (DO) consumption of Litopenaeus vannamei (13.2±1.42 g) juvenile maintained in
a microbial flock raceway system at high density (1 shrimp/liter). Each of two groups of 6 4-liter cylinders was placed either
in the dark or under light conditions of an enclosed greenhouse. The cylinders were filled out with raceway water and sealed
without receiving aeration. Four shrimp were assigned to each of three cylinders maintained in the dark and to three cylinders
maintained in the light. The remaining cylinders did not receive shrimp (control). Initial DO was determined for each treatment
(4.1±0.13 mg O 2 /L). Thereafter, DO content was determined every 10 min for a period of 30 minutes with a polarographic
digital oxygen meter. The oxygen consumption for shrimp maintained in the dark was 1.38±0.42, 1.08±0.12 and 0.55±0.28
mg O 2 /L for the 10, 20 and 30 min readings, respectively. The oxygen consumption for shrimp maintained under light conditions
was 1.14±0.37, 0.80±0.23 y 0.79±0.14 mg O 2 /L for 10, 20 and 30 minutes respectively. The oxygen consumption rate
was significantly different (P≤0.05) at 10 and 20 min between shrimp maintained in the dark and those maintained under light
conditions. When oxygen consumption was analyzed over time in presence of light, there were significant (P≤0.05) differences
between 10 and 20 min and between 10 and 30 min (Table 1). Under dark conditions there were only significant differences
in oxygen consumption between 20 and 30 min. It was concluded that anoxic conditions would be reached in less than 1 hour
either during the day (sunlight) or at night (dark) in the accidental absence of aeration/oxygenation.

THE NEAH BAY GEODUCK AQUACULTURE AND COMPARISON WITH GEOCHEMICAL
STUDIES IN HOOD CANAL
Yongwen Gao
Makah Fisheries Management
PO Box 115
Neah Bay, WA 98357 USA
gaoy@olypen.com
The Makah Fisheries Management (MFM) started a feasibility test on intertidal geoduck aquaculture in Neah Bay beaches
since 2006. To provide technical and scientific support for future commercial geoduck aquaculture in the area, MFM conducted
a geochemical monitoring project from the beginning of tube installation and systematically collected samples from seawater,
geoduck shell and flesh, and environmental parameters (e.g., temperature, salinity, dissolved oxygen, eelgrass, macro-algae,
and sand size and distribution) on a daily basis. Preliminary data showed that the 18 O/ 16 O variations were consistent with seawater
temperature changes in the geoduck beds whereas the Sr/Ca and Fe/Mn ratios were more related to water quality conditions.
The monitoring results appear in agreement with stable isotopic and trace elemental analyses on geoduck shell and seawater
samples from Hood Canal of Puget Sound. If successful, the comparison study may provide the first-hand information about
the geoduck growth, survival strategy, and ocean climate changes in both intertidal and subtidal settings. These geochemical
data and outcomes may also be helpful in estimating the population dynamics of wild geoduck stocks, and make a positive
contribution to the Washington geoduck aquaculture industry and scientific community.
11

11
GROWTH AND SURVIVAL OF Macrobrachium americanum Bate, 1868 JUVENILE PRAWNS
(CRUSTACEA, DECAPODA, PALAEMONIDAE) STOCKED IN TANKS AT DIFFERENT
SIZES
Marcelo U. García-Guerrero
Acuacultura de crustáceos
CIIDIR-IPN Unidad Oaxaca
Calle Hornos No.1003 C.P. 71230
Santa Cruz Xoxocotlan, Oaxaca
Apdo Postal 674 y 608
Oaxaca, Oax, Mexico
mgarciagu@ipn.mx
This essay appraises the growth and survival
of M. americanum juveniles, a prawn from
the west coast of America, based on the
effect of stocking size on wild specimens
maintained in tanks. Trial lasted 84 days
with a final survival from 55 to 72% depending
on their stocking size. Significant differences
in growth throughout the trial were
obtained.
Table 1. Survival (%) along time per group of
Macrobrachium americanum juveniles
stocked at different sizes and maintained
during 84 days. Treatment 1, 0.06 to 0.10 g;
Treatment 2, 0.11 to 0.20 g; Treatment 3,
0.21 to 0.30g. Different letters means
statistical differences among different
stocking sizes by one-way ANOVA (p≤0.05).
Treatment 1 2 3
Day
14 84.35a 91.74a 95.64a
28 66.52a 87.88b 92.01b
42 61.30a 76.03b 85.96b
56 58.70a 73.55b 77.72b
70 57.39a 69.97b 74.82b
84 55.22a 68.60b 72.40b
Table 2. Weight increase during the trial of
Macrobrachium americanum juveniles of the three
tested trials (different letters means statistical
differences within the same stocking size by one-way
ANOVA (p≤0.05). Trial 1, 0.06 to 0.10 g; Trial 2, 0.11
to 0.20 g; Trial 3, 0.21 to 0.30g.
Day/Trial 1 2 3
1 0.08±0.007a 0.15±0.012a 0.25±0.013a
14 0.17±0.014b 0.27±0.036b 0.48±0.071b
28 0.34±0.049c 0.41±0.074c 0.79±0.069c
42 0.67±0.071d 0.76±0.065c 1.25±0.095d
56 1.11±0.174d 1.24±0.098d 1.61±0.121e
70 1.90±0.271e 2.11±0.195e 2.26±0.184f
84 2.82±0.335e 3.26±0.214f 3.58±0.289g

POST-KATRINA MARINE BAIT DEVELOPMENTS FOR SOUTHEAST LOUISIANA
Albert P. Gaudé III
Louisiana Sea Grant Program/ LSU AgCenter
Louisiana State University
479 F. Edward Hebert Boulevard
Belle Chasse, Louisiana 70037 USA
agaude@agcenter.LSU.edu
With the catastrophic events that came with Hurricanes Katrina and Rita during the fall of 2005, virtually all marine bait infrastructure
was either heavily damaged or fully demolished. The slow pattern of recovery from these events has changed the
interwoven fabric of marine bait collection and distribution supporting the premier recreational fishing opportunities along the
entire southeast coast of Louisiana.
Pre-Katrina marine bait availability was based on a system of traditional bait harvesters, providing an erratic stream of live bait
shrimp and finfish to retail and private facilities that relayed items to recreational fishermen. The recreational marine fishing
industry in southeast Louisiana was experiencing an unprecedented expansion due to international recognition of the offshore
and bay populations of popular gamefishes. The marine bait industry was therefore struggling to keep pace with the quantity
and quality demanded by the increase in state and international fishermen. Historical marine bait facilities often relied on less
technical methods of harvest, transport, and holding systems.
With the rebuilding efforts after the events of August 2005, there has been a noticeable shift of the marine bait industry towards
newer technology and techniques. Accompanying this opportunistic upgrade has been increased financial investment and interest
in regulatory support for this critical component of a lucrative recreational fishing sector.
Further challenging the development of this marine bait industry has been the recent 2008 Hurricanes of Gustav and Ike.
Although not as destructive to the southeast Louisiana bait industry as Katrina/Rita, many bait harvesters and retail outlets
were temporarily disabled. In some cases, this ‘second punch’ in the midst of the rebuilding cycle has proven to permanently
overpower individuals involved in the industry. As always, some components have seen these events as opportunities rather
than setbacks.
11

11
TIMING OF FALL SCHOOLING BEHAVIOR AND HARVEST OPPORTUNITY OF BAITFISH
FAT SLEEPER Dormitator maculatus IN COASTAL LOUISIANA
Albert P Gaudé III
Louisiana Sea Grant Program/ LSU AgCenter
Louisiana State University
479 F. Edward Hebert Boulevard
Belle Chasse, LA 70037 USA
agaude@agcenter.LSU.edu
Of the many marine bait species in Louisiana, one of the most notorious and sought is the fat sleeper minnow Dormitator
maculatus. The reputation of this baitfish is legendary in its ability to entice predatory gamefishes to take bait and eventually
be landed by recreational fishermen. Perhaps part of this reputation is the short seasonal availability of this species in the retail
bait markets.
Locally known as ‘storm minnows’, this species is only commercially harvested in fall climatic events which combine tidal
surges and rainfall. The fish respond by having large gregarious schools which congregate at points of constriction in estuarine
waterways. These schooling events are temporally and spatially unpredictable. Further limiting their availability on the bait
market, these schools are ephemeral, often lasting for only a few days. The annual occurrence of the temporary schools can
vary by more than a month.
Since the marine bait industry has loosely documented private records, the appearance of this schooling behavior is poorly understood.
In an effort to assist develop this species as a bait candidate, peripheral survey information from state agencies were
examined to correlate the presence of this species with prevailing climatic conditions. Specifically, population net samples and
stomach contents analysis were used.
Normally, secretive and illusive to sampling efforts, the appearance in the sampled water column and stomach contents of
predatory fishes gives some index of availability to the bait industry. Although far from precise, this injects some temporal
predictability into the harvest sector for this species.

PHARMACOKINETICS OF FLORFENICOL IN CHANNEL CATFISH Ictalurus punctatus
Patricia S. Gaunt,* Christopher Wrzesinski, Cory Langston, Louis Crouch, Rodgers Polk, Dana Gao
and Richard Endris
Mississippi State University
College of Veterinary Medicine
Delta Research and Extension Center
P.O. Box 197
Stoneville, MS 38776 USA
gaunt@cvm.msstate.edu
Enteric septicemia (ESC) and Columnaris disease are the most serious bacterial diseases affecting farm-reared catfish (Ictalurus
punctatus) in the United States. Florfenicol is an antibiotic that was approved for use in catfish in 2005 for control of mortality
associated with ESC and in 2007 was conditionally approved for control of mortality associated with Columnaris disease.
The pharmacokinetics of intravenous and orally administered florfenicol (dose rate: 10 mg/kg once or 10 consecutive days)
were determined for catfish. At various time points ranging from 10 minutes to 36 hours after administration, blood was
sampled from individual fish (6 fish per time point).
The plasma was assayed for florfenicol using an LC-MS/MS method. The pharmacokinetic modeling of the results was performed
using the computer program WinSAAM. Data describing the distribution and elimination of florfenicol in catfish will
be presented.
11

120
EFFECT OF AMINO ACID DEFICIENCY AND FISH STRAIN ON PROTEIN DEGRADATION
SYSTEMS IN RAINBOW TROUT
T. Gibson Gaylord, Kenneth E. Overturf and Frederic T. Barrows
USDA-ARS Small Grains and Potato Germplasm Research Unit
Hagerman Fish Culture Experiment Station
3059F National Fish Hatchery Rd.
Hagerman, ID 83332 USA
Growth is known to be impaired due to amino acid deficiencies and is variable in different strains of trout. Alterations in degradation
pathways due to amino acid imbalances as well as variability between strains of trout have not been widely examined. In
order to assess changes in protein degradation systems, specifically the 20S proteosome and calpain systems, two experiments
were devised and carried out. The first experiment was designed to determine differences in protein degradation pathways
when diets were deficient in either methionine (13 vs. 19 g TSAA/kg diet) or lysine (12 vs. 29 g lysine/kg diet). The second
experiment investigated genetic differences between two strains of rainbow trout (R9 vs. House Creek) in the 20S proteosome
and calpain degradation systems when diets were either deficient or adequate in methionine.
In experiment 1, growth, feed consumption, feed conversion ratios, whole body crude protein and protein retention efficiencies
were negatively affected by deficiency of either amino acid. Feed consumption, whole body crude protein and protein retention
efficiency, were affected by both amino acid type as well as deficiency. Interactive effects of amino acid type and deficiency
on protein retention efficiency indicate that lysine deficiency reduced protein retention more than methionine deficiency. Feeding
diets with inadequate amino acid content increased the rates of both the calpain and 20S proteosome degradation systems.
Higher 20S proteosome activity was observed for the fish fed the lysine deficient diet than the fish fed the methionine deficient
diet.
In experiment 2, diet and strain both had significant effects on all growth and condition indices. Significant interactive effects
also were observed for growth (% gain and SGR), whole body crude protein, and protein retention efficiencies. Feeding methionine
deficient diets restricted growth potential for the House Creek strain to a greater extent than feeding the same diets to
the R9 strain.
The calpain and 20S proteosome degradation activities were elevated in fish consuming a methionine deficient diet compared
to the sufficient diet. No strain effects were observed on degradation activities of the two systems. However, there was a strain
effect on fish performance criteria.
In conclusion, it appears that there are differences in the regulation of the calpain and 20S proteosome protein degradation systems
based on which amino acid is limiting (lys vs. met, in experiment 1). While no strain effects were observed for the protein
degradation systems there appears to be differential utilization of amino acids for meeting the genetic potential of a trout strain
due to the observation that the House Creek strain had greater PRE when the diet was not limited by methionine.

DELTA 6 DESATURASE GENE IS DIFFERENTIALLY EXPRESSED IN THE INTESTINE
OF SEA BASS Dicentrarchus labrax FAMILIES EXHIBITING DIFFERENT GROWTH
POTENTIAL ONTO VEGETABLE-BASED-DIET
Florian Geay, Ester Santigosa, Thierry Tonon, Laurent Corcos, Pierre Boudry, Beatrice Chatain,
Jose Zambonino-Infante, Marc Vandeputte, Chantal Cahun and David Mazurais
Marine fish exhibit low HUFA (Highly Unsaturated Fatty Acids) synthesis capacity which represents a bottleneck for the substitution
of fish oils by vegetal oils in their feed. However, as some recent data revealed differential adaptability of fish to fish
oil replacement by vegetal oil, we assessed the regulation of the HUFA metabolic pathway in sea bass (Dicentrarchus labrax)
selected from two families (F1 and F2) that displayed similar growth rates when fed fish-based-diet (FD) but different ones
(F2>F1) when fed vegetable-based-diet (VD). The expression of the delta 6 desaturase (D6D) and elongase 5 (ELOVL5) genes
that encode two key enzymes involved in desaturation and elongation steps of HUFA synthesis were investigated in two major
metabolic organs, the liver and the intestine.
The switch from a FD to a VD induced an increase of D6D mRNA levels in the liver and intestine of both F1 and F2 fish-families.
The expression level of D6D was significantly higher in the intestine of F2 fish whatever diet considered (Figure 1). In opposite,
no difference in ELOVL5 expression was evidenced due to diets or families (Figure 2).Work in progress will determine
if the higher level of D6D expression in the intestine of F2 fish is related to an increase in the activity of desaturation and if this
is associated with a modification of fatty acids profiles in the muscle.
121

122
GEOGRAPHIC INFORMATION SYSTEMS (GIS) TRAINING FOR MUNICIPAL SHELLFISH
COMMISSIONS: Tools for Shellfisheries and Aquaculture Management
Tessa Getchis*, Michael Prisloe, Cary Chadwick, David Carey, Kristin Frank and Don Murphy
University of Connecticut
Sea Grant Extension Program
Groton, CT 06340-6048 USA
tessa.getchis@uconn.edu
Connecticut’s shellfish commissions, responsible for managing recreational and commercial shellfisheries within their town
waters, use several types of information in the evaluation of aquaculture applications including distribution and abundance
of local marine resources, mooring positions, shellfish classification areas, water quality data, and location of existing leases.
While much of this information is publicly available in Geographic Information System (GIS) data layers, these volunteer
municipal commissioners have not been exposed to formal training in the utilization of Global Positioning Systems (GPS), GIS
and geospatial data sets, and thus continue to rely on paper maps. These maps are often at different scales and are difficult to
overlay, which can result in confusion and delays in the permitting process.
Recognizing the need for better shellfisheries management tools, extension specialists developed a GIS training course for
municipal shellfish commissioners. Hands-on training sessions, using Environmental Systems Research Institute’s (ESRI)
ArcGIS software, introduced commission members to public domain geospatial datasets and to geospatial tools for collecting,
inputting, and editing geospatial data used for managing shellfisheries in their respective towns. Each commission was
assigned a field project to put their newly acquired information and skills to use following classroom training.
The goal of the project is to better equip shellfish commissions with the information and tools necessary to make informed
decisions regarding the siting of aquaculture activity including the establishment of new and/or expansion of existing leases, to
promote agriculture development in the state, and to manage local shellfish resources. The results and impacts of field projects
will be discussed.

INFLUENCE INDIVIDUAL AND MIXED HEAVY METALS (ZN, PB) AND DETERGENT
(LAS) ON THE MORTALITY FISH FRY 1G (Rutilus frisii kutum) OF CASPIAN SEA
Gholami, M.
Islamic Azad University
Sanandaj Branch, Iran.
Gholami62@yahoo.com
In this survey, the individual and mixed effects of heavy metals such as zinc and plomb and detergent (LAS) on 1g fish Fry
(Rutilus Frisii Kutum) were studied with 6 treatments and one blank in 3 replications using OECD method (1964) in the laboratory
and concentration limits were determined with logarithmic method for Plomb (0.11-0.17), zinc (0.1-0.5) and LAS (10-19)
mlg/l. Finally ,The achieved results were calculated with Probit analysis (Finny, 1971) and Correlation coefficient was regarded
for Plomb 0.93, zinc 0.92, and LAS 0.98 and in mixture of Plomb and LAS 0.93 and mixture of zinc and LAS 0.98.
The amounts of Lc 10 , Lc 50 and Lc 90 and permissible limits of above – mentioned pollutants were calculated too. Zinc effect on
fish fry caused Lc 10 =0.2 , Lc 50 =0.4 Lc 90 =0.7 ;Plomb effect caused Lc 10 =0.11, Lc 50 =0.21 , Lc 90 =0.34; LAS caused Lc 10 =5.91 ,
Lc 50 =11.62 , Lc 90 =22.71 ; mixture of LAS and Plomb caused Lc 10 =0.017 , Lc 50 =0.047 , Lc 90 =0.12 ; mixture of LAS and zinc
caused Lc 10 =0.02 , Lc 50 =0.09 , Lc 90 =0.37.
Hierarchically , the amounts of pollutants permissible limits for Plomb effect on fish fry were 0.021 , for zinc 0.4 , for LAS 1.16
and mixture of Plomb and LAS 0.004 and mixture of LAS and zinc 0.009.
123

124
ASSESSMENT OF INDIVIDUAL AND MIXED AMOUNTS OF TRANSMISSION OF HEAVY
METALS(CU,CD) AND (LAS)DETERGENT TO Rutilus frissi kutum FOOD CHAIN
Mansooreh Gholami and Maryam Fallahi
Islamic Azad University
Sanandaj Branch, Iran
Gholami62@yahoo.com
In Assessment of Individual and Mixed amounts of Transmission of Heavy Metals(Cu,Cd) and (LAS)detergent to Rutilus
frissi kutum food chain ,the results showed that the transmission of heavy metal(Cadmium) to food chain from the amount
of permissible limit added to Scenedesmus obliquus culture medium with amount of 0.0127 ppm, renged from 0.0041 in the
polluted Scenedesmus obliquus to 0.0015 in Daphnia magna ,while this amount was not significant in Rutilus frissi kutum fed
on polluted Daphnia magna .This amount in Rutilus frissi kutum fed on polluted Scenedesmus obliquus was 0.002 ppm and
the amounts of transmission in food chain were\calculated as 32%,8% and 16% Copper heavy metal transmission from the
amount of permissible limit added to Scenedesmus obliquus culture medium with amount of 0.15 ppm was 0.07 in polluted
Scenedesmus obliquus ,0.02 in Daphnia magna and insignificant amount in Rutilus frissi kutum .In Rutilus frissi kutum fed
on polluted Scenedesmus obliquus,the amount of 0.02 was achieved and transmission to food chain was 46%, 13% and 13%.
The transmission amount of (LAS+Cd) from the amounts of pollutants permissible limits added to Scenedesmus obliquus culture
medium with the amount of 2.153 and 0.0127 ppm were 0.007 in polluted Scenedesmus obliquus and 0.003 in Daphnia
magna ,while this amount in Rutilus frissi kutum fed on Daphnia magna reached to 0.001 ppm and in Rutilus frissi kutum fed
on polluted Scenedesmus obliquus reached to 0.003 ppm and the amounts of transmission to the food chain were 58%,25%,8%
and 25%.moreover,the achieved results from the transmission of(LAS+Cu) mixture showed that from the amounts of pollutants
permissible limit added to Scenedesmus obliquus culture medium with amount of 2.153 and 0.15 ppm were 0.09 ppm in polluted
Scenedesmus obliquus.The amounts of transmission in food chain ranged as:60%,33%,6% and 45%.The results showed
that the percentage of Cu and Cd transmission in to food chain is of a significant amount individually or mixed with LAS

FORMALIN TREATMENT FOR SAPROLEGNIA INFECTION IN CHANNEL CATFISH
Charles M. Gieseker*, Christie-Sue Cheely and Renate Reimschuessel
United State Food and Drug Administration
Center for Veterinary Medicine
Office of Research
8401 Muirkirk Road
Laurel, Maryland 20708 USA
Aquatic fungi can cause serious infection and mortality in cultured catfish, especially following cold spells. We conducted a
controlled GLP study to determine if formalin can reduce mortalities associated with such infections. Disease was induced in
Channel catfish, (Ictalurus punctatus) by immersing fish in an exposure tank seeded with the aquatic fungi, Saprolegnia parasitica
(ATCC 22284) after a controlled epidermal abrasion. Fish were then randomly distributed into 16 experimental tanks (3
fish/tank) held at 14±2 ºC to simulate the stress induced by cold weather. Fish were treated with formalin (0, 50, 100, and 150
ppm) 1 hour after being placed into the experimental tanks (day 1) and on days 3 and 5. Fish were monitored for 19 days. All
doses of formalin significantly reduced mortality compared to the control fish, (control=68%, 50 ppm -50%, 100 ppm, 32%,
150 ppm=38%) p

12
USING NEAR-INFRARED REFLECTANCE SPECTROSCOPY (NIRS) TO PREDICT THE
DIGESTIBLE PROTEIN AND ENERGY VALUE OF GRAIN INGREDIENTS WHEN FED TO
RAINBOW TROUT, Oncorhynchus mykiss - THE LUPIN CASE STUDY
Brett Glencross*, Wayne Hawkins, Peter Burridge, David Evans, Neil Rutherford, Peter McCafferty, Ken Dods
and Sofia Sipsas
Department of Fisheries
PO Box 20, North Beach
WA 6920, Australia
Brett.Glencross@fish.wa.gov.au
Over a five-year period, 10 separate digestibility experiments undertaken to examine the digestibility of protein and energy
from 136 different samples of lupin meal from either Lupinus angustifolius or L. luteus. Lupin samples were obtained from the
Australian National Lupin Breeding Program’s germplasm lines and selected on the basis of maximal crude protein variability
as assessed by existing crude protein NIRS calibrations. Chemically measured crude protein values varied from 232 to 613 g/kg
DM. Gross energy values ranged from 18.7 to 21.6 MJ/kg DM. Other compositional parameters assessed included amino acids,
total lipids, ash, total carbohydrates, cellulose, hemicellulose and lignin.
The digestibility of protein and energy was assessed using the diet-replacement ingredient assessment method, where the test
ingredient comprised 30% of each test diet. Digesta was collected using faecal stripping techniques. Digestible protein values
ranged from 193 to 595 g/kg DM and digestible energy values ranged from 6.0 to 17.7 MJ/kg DM. The digestible protein and
energy values were then assessed using multiple regression techniques to determine which compositional parameters accounted
for the majority of the variability. Crude protein content was observed to be the most dominant single factor in affecting digestible
protein levels. However, multiple regression analysis supported that protein and lignin content combined were the strongest
predictors of digestible protein value, explaining close to 60% of the variability in this parameter. Digestible energy values
were hevily reflective of the factors influencing digestible protein value. This study demonstrates that within one raw material
type that not only does significant variability in the digestible value of the raw materials exist, but that it is possible to identify
compositional features of that raw material that are intrinsically influencing its own digestible value. Predictive equations have
been developed that explain this variability.
Assessment of these samples using near-infrared spectroscopy (NIRS) shows that there are certain wavelengths that correlate
with crude protein and lignin content of the lupin kernel meals. Applying a dual assessment to both of these parameters, based
on the predetermined relationship, it has been possible to develop a calibration to predict both digestible protein and energy
content of lupin kernel meals. This calibration can now be used to assess lupin meals prior to formulation to minimise digestible
protein and energy content of diets in which they are included.

APPLICATION OF SCALE COVER GENE N FOR IDENTIFICATION OF PLOIDY IN
ORNAMENTAL (KOI) CARP
Boris Gomelsky*, Kyle J. Schneider and Robert P. Glennon
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
boris.gomelsky@kysu.edu
Production and stocking of sterile triploid ornamental (koi) carp might be useful in cases when fish reproduction is undesirable.
The common way to produce triploid fish is suppression of the 2 nd meiotic division in eggs by application of strong physical
treatments (temperature shocks or hydrostatic pressure). Usually applied physical treatments do not provide 100% triploids in
progenies; therefore the ploidy of each fish should be verified by analysis of blood sample. It would be beneficial to develop a
method of ploidy determination based on some external morphological trait.
Appearance of four scale cover types (scaled, mirror, linear or leather) in koi (and common carp) results from the interaction of
two genes having two alleles each (S/s and N/n). Experiments on induced meiotic gynogenesis revealed a very high recombination
rate of gene N; almost 100% of gynogenetic fish resulting from a heterozygous female (Nn) are heterozygous. This study
suggested using this peculiarity of gene N to determine fish ploidy.
Eggs taken from a leather koi female (genotype ssNn) and sperm taken from a scaled koi male (SSnn) have been used for
production of control (no shock) and heat-shocked progenies. The 2-min heat shock (40º C) was applied 6 min after insemination.
Fish segregation in control progeny (288 scaled : 315 linear) did not differ significantly from the expected ratio 1:1. Fish
segregation in heat-shocked progeny (179 scaled : 404 linear) was significantly shifted towards the prevalence of linear fish.
This shift in ratio may be explained by the suggestion that due to high recombination frequency of gene N, the suppression of
2 nd meiotic division results mainly in triploids with genotypes Nnn, while scaled fish nn should be mainly diploid. In heatshocked
progeny about half of linear fish had less profound reduction of scale cover compared with typical linear fish. It was
suggested this group (called multi-scaled linear) are triploid fish (Nnn) which differ from typical diploid linear fish (Nn) due
to incomplete dominance of allele N.
These suggestions have been generally confirmed by analysis of fish ploidy based on measurements of erythrocyte nuclei by a
Coulter counter. All analyzed (n=20) scaled and typical linear fish from heat-shocked progeny were diploid while from 15 analyzed
multi-scaled linear fish 14 fish were triploid and 1 fish was diploid (Figure 1). The practical applicability of this method
of ploidy determination will be discussed.
12

12
ASSESSING THE HEALTH OF RAINBOW TROUT Oncorhynchus mykiss IN LOW-
EXCHANGE RECIRCULATING SYSTEMS OPERATED WITH OR WITHOUT OZONATION
Christopher Good*, John Davidson, Carla Welsh and Steven Summerfelt
The Conservation Fund’s Freshwater Institute
1098 Turner Road
Shepherdstown, WV 25443 USA
c.good@freshwaterinstitute.org
Previous observations by the authors have indicated that rainbow trout Oncorhynchus mykiss mortality increases sharply in
recirculating aquaculture systems (RAS) when water exchange rates are lowered to the point where feed loading rates are approximately
1.3-2.0 kg/d per m 3 /d of make-up water flow. All measured water quality parameters were within safe limits at the
time, and no specific cause of mortality was ascertained. Ozonation of the water appeared to remediate the fish health decline.
Subsequent research at The Freshwater Institute has focused on examining environmental conditions and fish health in low
exchange RAS with high feeding rates, as the previously observed mortality phenomenon could represent an important barrier
for economic sustainability in commercial recirculating aquaculture.
The objective of the present study was to evaluate the effects of water ozonation on fish health and performance in RAS with
low exchange and high feeding rates. Six replicated RAS, all operated at low exchange rates (0.26% of the total recirculating
flow), were each stocked with 1000 rainbow trout, and densities were maintained between 40 and 80 kg/m 3 for the entire study
period. Additionally, three small tanks in a flow-through system each received 100 rainbow trout, and were fed and maintained
under similar conditions as a physiological comparison group. Three of the six RAS were randomly selected to receive water
ozonation (to an oxidation-reduction potential of 250mV), while the remaining three RAS did not receive ozonation. Average
fish weight across all systems was approximately 300 grams at the time of treatment initiation. Fish were subsequently raised
for four months under these conditions; during the first month after ozone addition, tanks were fed equal portions to investigate
water quality and unit process treatment efficiency differences, after which each tank was fed to satiation.
Fish health and performance were assessed with daily mortality and monthly length and weight data collection, as well as fin
index evaluation, gill, liver and ovary minerals and metals content screening, blood gas and blood chemistry analyses, and multiple
tissue sampling for histopathological assessment, all carried out during the final week of the study period. By study’s end,
there was no significant (p

THE EFFECTS OF HIGH VS. LOW DISSOLVED CARBON DIOXIDE ON THE HEALTH
AND PERFORMANCE OF RAINBOW TROUT Oncorhynchus mykiss IN LOW-EXCHANGE
RECIRCULATING AQUACULTURE SYSTEMS
Christopher Good*, John Davidson, Carla Welsh, Kevin Snekvik and Steven Summerfelt
The Conservation Fund’s Freshwater Institute
1098 Turner Road
Shepherdstown, WV 25443 USA
c.good@freshwaterinstitute.org
Previous un-replicated experiments at The Freshwater Institute have indicated that rainbow trout Oncorhynchus mykiss mortality
increases with low water exchange rates during feed loading of 1.3-2.0 kg/d per m 3 /d of make-up water flow in recirculating
aquaculture systems (RAS). No infectious diseases or opportunistic infections were diagnosed at the time, and typical water
quality parameters were within safe limits. This mortality spike could represent an important barrier for producers raising rainbow
trout in RAS with low exchange rates. Subsequent efforts to reproduce and evaluate this fish health decline in controlled
experiments have not succeeded; however, dissolved CO 2 levels during these controlled studies were approximately 40% of
CO 2 levels during the original mortality episode. Because exposure to elevated CO 2 is associated with many physiological
disturbances in fish, further experimentation was warranted to study the effects of elevated CO 2 in low exchange RAS on fish
health and performance.
This study compared rainbow trout health and performance between three RAS operated with high CO 2 levels (25 mg/L) and
three RAS with low CO 2 levels (10 mg/L). All systems were operated at low water exchange rates (0.26% of the total recirculating
flow). Rainbow trout (64 ± 1 grams) were randomly stocked into the six RAS (800 fish per system), and 100 fish were
stocked into three small tanks within a flow-through system to provide a physiological comparison group for the RAS cohorts.
A 24-hr photoperiod was provided, and all tanks were fed equal portions during the six-month study period.
Fish health and performance were assessed with daily mortality and monthly length and weight data collection, as well as multiple
tissue samplings (skin, skeletal muscle, gill, heart, liver, spleen, pyloric cecae, swim bladder, and anterior and posterior kidney)
for histopathological assessment. At study’s end, the previously observed fish health decline had not been reproduced, and
percentage survival for both groups was excellent (>97%). No significant (p

130
OKLAHOMA’S PADDLEFISH MANAGEMENT AND CAVIAR PROGRAM
Gordon, Brent D.
Oklahoma Department of Wildlife Conservation
N.E. Regional Fisheries Supervisor
P.O. Box 1201
Jenks, Oklahoma 74037 USA
bgordon07@sbcglobal.net
Oklahoma has one of the largest naturally-reproducing paddlefish (Polyodon spathula) populations in North America and offers
what are arguably the best recreational fishing opportunities for these fish anywhere in the world. The current paddlefish
fishery is the result of a concerted management by Oklahoma Department of Wildlife Conservation (ODWC) biologists and
rigorous anti-poaching efforts by ODWC’s game wardens. With such a good fishery, agency personnel saw both a challenge
and opportunities on the horizon.
To improve the management and conservation of this species, ODWC personnel saw a need for additional studies examining
the long-term effects of threats such as the introduction of exotic species, illegal harvest, and changes in the watersheds where
paddlefish are found are necessary. Some of the biological data that were needed cannot be obtained without sampling large
numbers of fish. The agency does not want to kill fish just for the sake of a research project, and until this program, there was
no large-scale system for collecting data from angler-caught fish.
The approach taken by ODWC consisted of installing a Paddlefish Research and Processing Center (RPC) and the equipment
needed for the collection of data. On a voluntary basis, 4,221 legally harvested paddlefish were brought to the RPC where
basic biological information such as body length, weight, sex, condition, gonadal weights (green eggs), and gonadal fat weights
were recorded. Dentaries were also collected from a high fraction of fish brought into the station. The number of paddlefish
needed to obtain statically valid data cannot feasibly be obtained in any other situation. In this program, sport anglers donate
paddlefish eggs in exchange for having their fish cleaned and bagged free of charge. The roe are then processed into caviar
and sold on the world market.
In the first year of operation the RPC produced 3,636kg of paddlefish caviar. Implementation of this program should give the
ODWC the necessary information for assessing stock status and determining acceptable harvest levels. The project is designed
to provide a framework for proper stock assessment, monitoring, and management as part of a recreational paddlefish fishery.
The main purposes of the program are to: 1) provide alternative funding sources for continued monitoring and expanded research;
2) utilize a resource currently being wasted (discarded roe). 3) facilitate the gathering of biological data such as dentary
bones for determining population age structure and gonadal information from paddlefish to properly answer questions about
reproduction and recruitment in Oklahoma and 4) to increase the Oklahoma Department of Wildlife’s law enforcement capabilities,
which will help reduce illegal commercialization of paddlefish and eggs.

SEA TRIALS OF A SELF-PROPELLED OFFSHORE SEA CAGE
Clifford A. Goudey*, Stephen H. Page and Brian C. O’Hanlon
Offshore Aquaculture Engineering Center (OAEC)
Massachusetts Institute of Technology Sea Grant College Program
MIT Building E34-356
Cambridge, MA 02142 USA
The U.S. Exclusive Economic Zone (EEZ) is vast, exceeding the land area of the nation, and holds the potential for inestimable
production of seafood and other cultured products. Yet current technologies available for industrial-scale aquaculture are
moored systems and their use is restricted to a narrow portion of coastal waters that are within serviceable depths for anchoring
but are often the most contentious parts of the ocean. By contrast, the deeper portions of the EEZ have fewer users, better water
quality, and currents tend to be consistent and predictable, being driven by large-scale ocean circulation processes.
An offshore culture system that is un-tethered could enjoy numerous advantages including reduced cost, more consistent water
temperatures and water quality, and relief from certain regulatory impediments that are currently constraining the growth of
aquaculture. Model tests of a self-propelled fish cage have previously demonstrated the feasibility of the concept. A recent
project funded by the NOAA Marine Aquaculture Program has supported the development and demonstration of such mobile
cage technology at commercial scale.
A self-propulsion system was designed and installed on an Ocean Farm Technologies AquaPod 3,250 cubic meter cage. This
cage is a geodesic sphere designed for submerged operation and constructed of modular panels made of a composite polymer
frame and welded, galvanized, and PVC-coated wire mesh that is resistant to predation. The structural integrity of the AquaPod
made it an ideal candidate for self-propelled operations.
To propel the cage we used two 2.5 m. diameter electric thrusters featuring two-bladed, highly skewed propellers that produce
92 kg of thrust per kW or 151 pounds per hp. See Figure 1.
These units were controlled by a pair of variable frequency motor controllers, which we powered by a diesel generator set
aboard a boat towed by the cage.
Our experiments were conducted during June 2008 at Snapperfarm, a commercial fish farm in Culebra, Puerto Rico, and included
speed trials at various power levels, maneuverability tests, and tests to determine our ability to maintain a straight-line
course. The results of these tests are reported and provide a firm basis for continued research on the mobile cage operations.
The tasks accomplished in this project represent an essential step in the methodical development of fish farming in the U.S.
EEZ as well as broader areas of the oceanic basins.
Figure 1. The thruster pair in operation
131

132
PRODUCTION CHARACTERISTICS OF CHANNEL CATFISH AND CHANNEL X BLUE
CATFISH IN EARTHEN PONDS
Bartholomew Green
USDA-ARS Harry K. Dupree Stuttgart National Aquaculture Research Center
PO Box 1050
Stuttgart, AR 72160 USA
bart.green@ars.usda.gov
The channel (female) x blue (male) hybrid catfish increasingly is being selected by catfish farmers in the United States as their
culture fish. This completely randomized design study in 2 x 2 factorial arrangement was conducted to compare production
traits in earthen ponds of the channel x blue hybrid catfish to those of channel catfish from the same line used to make the
hybrid catfish when fish were fed full and restricted rations. Juvenile catfish were stocked into four replicate 0.1-ha earthen
ponds per treatment at 14,820 fish/ha during March-April. Each pond was equipped with a 0.559 kW electric paddlewheel
aerator that was turned on when pond dissolved oxygen was less than 4.5 mg/L. Fish were fed daily as much 32% protein feed
as they could consume in a 20-min period. Fish within each genetic group in the restricted feed treatment were fed 80% of the
mean amount of feed consumed by fish in the unrestricted feed treatment. Water quality in ponds was monitored throughout
the experiment. Ponds were harvested in November. The effects of genetic line and feeding rate on fish production and pond
water quality were evaluated.

DEVELOPMENT OF A VALUE-ADDED PRODUCT USING CHANNEL CATFISH BELLY
FLAP MEAT
Bartholomew Green*, Byungrok Min, Lee Wiles and Jin Kim
USDA-ARS Harry K. Dupree Stuttgart National Aquaculture Research Center
PO Box 1050
Stuttgart, AR 72160 USA
bart.green@ars.usda.gov
The shank fillet of channel catfish is a primary product form produced during processing and is formed by trimming the belly
flap or nugget section from a boneless fillet. Belly flap meat comprises around 20% of the fillet weight. Belly flap meat has a
fat content of about 15% compared to about 9% for the shank fillet. The fat content of the belly flap meat, which limits its shelf
life and confers undesirable sensory qualities, along with the black-pigmented peritoneal membrane present on the surface of
the belly flap contribute to its classification as a low-value by-product of catfish processing. Wholesale price paid for catfish
belly flap meat, also known as catfish nuggets, averages about 37% of the shank fillet price.
A series of studies was undertaken to evaluate the potential for developing a value-added product using minced channel catfish
belly flap meat. Washing of catfish mince resulted in whiter tissues or products due to the removal of blood, pigments, and
water-soluble proteins; however, the presence of specs of comminuted peritoneal membrane resulted in grayish or dark areas
on certain points of the product. Washing increased the moisture content from 71.5% to 77.9% while reducing mechanical
hardness (19.1 N to 11.9 N), chewiness index (14.2 N to 8.83 N) and shear force (0.82 J to 0.51 J), and fat content (9.82% to
4.39%). The particle size of minced belly flap meat affected significantly textural characteristics of catfish patties cooked by
dry or moist heat. As size of meat particles for patties decreased, hardness and gumminess, and color difference from fillet
increased, but Kramer shear break force as well as percent cooking loss decreased (P

134
REPRODUCTIVE OUTPUT FOR COCAHOE MINNOW Fundulus grandis IN ABOVE
GROUND POOLS AT DIFFERENT STOCKING DENSITIES
Christopher C. Green, Craig T. Gothreaux and Greg C. Lutz
Aquaculture Research Station
Louisiana State University Agricultural Center
2410 Ben Hur Rd.
Baton Rouge, LA 70820 USA
cgreen@agcenter.lsu.edu
Live bait is one of the most popular fishing methods for recreational anglers; however, the marine baitfish aquaculture industry
is small compared to the multimillion dollar freshwater baitfish industry. A 2004 survey showed that 47% of Louisiana coastal
anglers considered the inconsistent availability of live bait to be a significant problem. This survey further indicated that the
cost of live bait (cocahoe minnows averaging $2 /dozen at the time) was not an important problem, justifying rapid development
of techniques to serve this market. Production of cocahoe minnows (Fundulus grandis) in above ground pools has a
number of potential advantages. A greater level of control on a captive population serves to better control many variables that
are inherent in pond culture. The objective of this research was to investigate the practical application of static above ground
pools for the production of cocahoe minnow, specifically: 1) egg output over time, 2) effect of spawning substrate, and 3) effect
of stocking density.
On March 12, 2008, 2000-L above group pools were filled with pond water and maintained at a salinity of 5 mg/L with constant
aeration throughout the spawning season. There were a total of 18 pools, half receiving cured Spanish moss spawning substrate
and the others receiving manufactured spawning substrate (Spawntex). Triplicate groups of cocahoe minnows were stocked at
three densities at a rate of 30, 60, and 90 per pool, with a sex ratio of 2:1 females to males, and fed 3.5 % body weight per day.
Each pool received a spawning mat (35 cm x 46 cm) immersed approximately 8 cm below the surfaced of the water. Mats were
replaced twice per week, and eggs were removed and counted to assess percent fertilization and egg size. Temperature data
loggers were deployed at the beginning of the season and recorded temperature throughout the study. Gonadosomatic index
(GSI) for males and females was assessed monthly with a separate group of individuals.
The Spanish moss substrate was significantly less productive than the Spawntex mats. Egg production peaked in May with
average pool temperatures of 27.4 + 3.1 o C. During May individual egg output from the three stocking densities ranged between
10 and 75 eggs every three days. Individual egg output per week does appear to differ among treatments densities and will be
conclusively investigated upon completion of the spawning season. It is anticipated that both GSI and monthly egg production
will follow temperature changes throughout the season. At the time of abstract submission egg production has not ceased and
it is anticipated that egg production will continue into the fall. The effect of stocking densities, temperature, photo and lunar
periods on egg production and GSI will be reported.

THE EMERGENCE OF VIRAL HEMORRHAGIC SEPTICEMIA VIRUS TYPE IVB IN THE
LOWER GREAT LAKES BASIN
Geoffrey H. Groocock*, Stephen A. Frattini, Geoffrey E. Eckerlin, Rodman G. Getchell, Gregory A. Wooster,
William T. Heath, Kristine M. Hope, Rufina N. Casey, John M. Farrell, James W. Casey and Paul R. Bowser
Aquatic Animal Health Program
Department of Microbiology and Immunology
College of Veterinary Medicine
Cornell University
Ithaca, New York 14853 USA
This presentation will present a timeline of events from the initial identification of viral hemorrhagic septicemia virus (VHSV)
type IVb in the Lower Great Lakes basin to the current extent of the disease in New York State waters. The emergence of
VHSV in New York State has provided more insight into the spread of a new pathogen into a naïve environment.
The first detection of VHSV in New York State waters occurred in May 2006. The virus was isolated and identified from a
mortality event of round gobies collected from the St. Lawrence River. The initial clinical signs displayed by these gobies and
resulting gross and histopathology will be presented.
Subsequent isolations of VHSV in 2006 included 21 different mortality or morbidity events from 16 locations, including the
Great Lakes, the St. Lawrence River and the first inland isolation in Conesus Lake. An initial surveillance effort among healthy
fish from the St. Lawrence River also detected VHSV in an additional 12 species.
In 2007 VHSV was diagnosed in a total of fourteen cases. These cases identified VHSV in the following new species in New
York State: gizzard shad, carp, rainbow trout and lake trout. New locations for 2007 included more locations along Lake Ontario
and Lake Erie, Skaneateles Lake, Seneca-Cayuga Canal, Little Salmon River and a private pond in Ransomville.
Detections of VHSV in 2008 were limited to isolations of the virus from round gobies. The impact of VHSV and the potential
future implications of this emerging pathogen will be discussed.
13

13
INVESTIGATION OF SPECIES SUSCEPTIBILITY TO VHVS TYPE IVB AND
INVESTIGATION OF THE USE OF IODINE EGG-DISINFECTION ON VHSV
Geoffrey H. Groocock*, Stephen A. Frattini, Rodman G. Getchell, Gregory A. Wooster, William T. Heath,
Kristine M. Hope, Rufina N. Casey, Steven R Lapan, Andrew D. Noyes, James W. Casey and Paul R. Bowser
Aquatic Animal Health Program
Department of Microbiology and Immunology
College of Veterinary Medicine
Cornell University
Ithaca, New York 14853 USA
The effect of VHSV in the Great Lakes may have a significant potential impact on aquaculture and recreational fishing species
such as channel catfish, walleye and muskellunge. Currently there is little information known on the relative susceptibility
of VHSV type IVb on these different species. Infection trials were performed on each of these species. Fish were inoculated
using the type Michigan 2003 isolate by immersion in water containing a known concentration of virus for one hour or by direct
intracoelomic injection of a small volume of known virus concentration. Control fish were sham inoculated using a sterile
sample of the media used to propagate the virus. Initial results of these experiments will be discussed.
Additionally, the effects of topical egg disinfectants such as iodine on VHSV are unknown. A trial experiment to determine
whether VHVS can be detected in eggs after exposure to VHSV containing water was performed. The experiment included
varying doses of VHVS, different concentrations of iodine disinfection and different methods of reducing egg adhesiveness
using tannic acid or Fuller’s earth. The result of the trial experiment will be presented and the proposed full experimental trial
will be discussed.
BIOACCUMULATIVE CONTAMINANTS IN PADDLEFISH IN LARGE RIVER SYSTEMS OF
THE UNITED STATES
Deke Gundersen and Steven D. Mims
Environmental Science Program
Pacific University
Forest Grove, OR 97116 USA
deke@pacificu.edu
Paddlefish inhabiting impounded large river systems that are influenced by industrial and agricultural processes are susceptible
to bioaccumulation of persistent bioaccumulative contaminants. Contaminants that are a particular problem in many of these
large river systems throughout the United States include chlorinated hydrocarbons (i.e PCBs, DDT, chlordane, and dioxins) and
methyl mercury. Many studies indicate that these contaminants negatively impact the health of paddlefish populations inhabiting
these large river systems. In addition, the chemical properties of these contaminants allow them to bioaccumulate in the
edible tissues of these species. This can potentially pose health risks for people consuming edible products from these species
(roe and meat), particular certain susceptible individuals (pregnant women and young children). This paper will review previous
studies investigating bioaccumulative contaminants in paddlefish in large river systems of the United States and compare
to farm-raised paddlefish as well as the conditions in these large rivers that make these long-lived species susceptible to uptake
of these chemicals.

COMPREHENSIVE QUALITY ASSESSMENT OF ATLANTIC COD (Gadus morhua L.)
FARMED UNDER DIFFERENT LIGHT REGIMES
Ørjan Hagen* and Christel Solberg
Faculty of Biosciences and Aquaculture
Bodø University College
NO-8049 Bodø, Norway
In Norwegian aquaculture, Atlantic cod is considered to be the next rising star due to it world wide popularity and so far promising
industrial results. To date, one of the largest challenges in the grow-out phase is high production cost due to precocious
maturation and slow growth. Superimposed on this, quality problems during the summer, particularly soft texture and gaping,
makes harvesting a “risky” business. This is unfortunate since the concept of fish farming is based on whole year harvest, and
particularly in periods when the price and demand is good (e.g. the summer).
Light manipulation as a mean to delay maturation and stimulate growth has not yet been very successful in large scale cod farming.
At the most, maturation has only been delayed with up to 6 months. One potential way of reducing the quality problems
during the summer is to apply existent light technology on both accelerated and delayed maturation. This would ensure that
netpens exposed to different light regimes matures at different times of the year, making harvesting of sub populations possible
during the whole year and therefore limit the fillet quality problems in the summer. To test this theory, a large scale experiment
was conducted outside Bodø (Norway) at a commercial farm site. In the present experiment (01.08.07–20.06.08) a total of five
netpens were included (n=60.000 each). Four netpens received a light regime causing delayed (DG) or accelerated maturation
(AG) respectively (two each), and one netpen contained the control group (CG, ambient conditions). Growth and maturation
status was monitored by sampling 30 individuals from each netpen from 05.12.07–20.06.08 (n=4). By the end of the experiment,
both AG and DG had outgrown CG with > 0.8 kg (P

13
ENERGY EFFICIENCY IN ALLIGATOR CULTURE OPERATIONS
Steven G. Hall*, Mark Shirley, Rebecca Schramm and Milton Saidu
Biological and Agricultural Engineering
LSU AgCenter
Baton Rouge, LA 70803 USA
sghall@agcenter.lsu.edu
American alligator (Alligator mississippiensis) culture has grown in recent decades, with Louisiana and Florida being the
largest culture states. Among the unique requirements for these reptiles is water as a habitat, a high protein diet, and a strong
linkage between culture temperature and growth rate. Many alligator culturists attempt to maintain culture temperatures in the
range of 85-90F (~30-32). These temperatures are fairly easy to maintain during the summer in southern states, but require
significant energy input in cooler seasons. These culture conditions reduce from 4-5 years to 11 months the time required to
grow out alligators to 4 feet (1.1meters), a typical sales size.
This project focused on ways to reduce energy costs, including use of ground loop heat exchange with warm waste lagoons, use
of solar energy for preheating or supplemental heating, and consideration of other techniques to reduce energy requirements.
Preliminary results suggest the possibility to reduce energy costs in the range of 50% for areas of focus. Future work should
consider other areas to save energy on these unique farms.
OFFSETTING CARBON EMISSIONS VIA ENGINEERED OYSTER REEF CULTURE
Steven G. Hall
Biological and Agricultural Engineering
LSU AgCenter
Baton Rouge, LA, 70803 USA
sghall@agcenter.lsu.edu
Combustion of fossil fuels for energy, unsustainable use of biomass, volcanic and other geological activity, each produce net
emissions of carbon dioxide, a recognized greenhouse gas. A number of mechanisms have been explored or proposed to reduce
or reverse carbon emissions, including capturing carbon chemically, placing carbon in areas less accessible to the atmosphere,
using plant biomass to temporarily sequester carbon and simply reducing fossil fuel use. This project explored the possibility of
using oyster reefs in sedimentary coastal areas to sequester carbon, primarily in the form of oyster shell (significantly composed
of calcium carbonate). Estimates of fossil fuel use were compared to growth and sedimentation rates for oyster reefs, based on
preliminary biological and engineering experiments on engineered oyster reefs.
Although the scale would be massive, it appears that these reefs could sequester carbon at a rate sufficient to offset carbon
emissions from fossil fuel use in the state of Louisiana, and could be a significant technique for overall carbon sequestration.
Additional considerations include subsidence rate (approximately one centimeter per year at present in areas of coastal Louisiana),
which would allow reefs to subside and thus store this carbon longer term (centures-millenia). Another consideration is
both ecological benefits (e.g. juvenile fish habitat, coastal protection, potentially enhanced oyster fisheries), as well as environmental
costs and benefits. For example, the initial material is expected to have some carbon costs which would be recovered
over time, the initial substrate must be accessible to oysters, structurally sound, but also environmentally friendly. Shipping
concerns and the effects (likely positive) on shoreline protection are also areas of interest. Although these numbers strongly
suggest that overall fossil fuel use must be reduced, it also shows that this is a realistic possibility for long term carbon sequestration
in an environmentally friendly and cost effective technique.

LIVE BAIT SHRIMP MARKETING IN THE GULF OF MEXICO AND SOUTH ATLANTIC
STATES
Terrill R. Hanson* and Ben Posadas
Department of Aquaculture and Allied Aquacultures
Auburn University
203 Swingle Hall
Auburn, Alabama 36849 USA
trh0008@auburn.edu
A live bait shrimp market survey instrument was developed and sent out in May/June 2008. The goal of the study was to better
understand the supply and demand of live bait shrimp at the recreational outlet level along the Gulf of Mexico and South
Atlantic states. Results of this survey effort will be presented.
The survey was sent to 543 marine bait dealers whose addresses were found through searching related internet websites. Fiftynine
useable surveys were returned, 26 returned surveys stated they did not sell live bait shrimp and 3 replied saying they had
gone out of business. There were 124 undeliverable surveys (but returned to us via USPS); and 331 no responses. Thus, there
were 390 potential surveys and a response rate of 15.1%.
Survey results are expected to provide information on the amount of live bait shrimp sold and demanded from live bait shrimp
suppliers. Additionally, the survey was designed to obtain information that will be needed by aquaculture operations in producing
native live bait shrimp species (white shrimp Litopenaeus setiferus and pink shrimp Farfantepenaeus duorarum).
Examples survey question areas and their anlaysis include:
- Size of shrimp preferred by dealers (and their customers): important in developing the shrimp culture facility, production
management, and timing (also cost of production will differ by size required);
- Potential quantity of shrimp required within a region: will provide valuable information regarding the size of hatchery and
production system required to partially meet this demand;
- Time of year live bait shrimp are needed: will give aquaculturists information on how they must hatch, grow and/or maintain
(store) live shrimp sizes for availability around peak demand times of the year;
- Prices for live bait shrimp: prices bait shop operators have paid and would be willing to pay for captured versus cultured bait
shrimp; will determine whether cultured shrimp characteristics, such as greater availability, better appearance, suitable sizes,
less mortality, etc., will evoke a premium price from bait dealers or charter boat captains;
- Additional information: how long bait store has been in business; number of holding tanks on-site, present source of bait;
shrimp species purchased; monthly quantities needed; retail sales price; and factors determining from whom they buy.
13

140
COST OF PRODUCING STOCKER AND HARVEST SIZE CATFISH Ictalurus punctatus IN
COMMERCIAL-SCALE VERIFICATION TRIALS
Terrill R. Hanson*, Louis R. D’Abramo, James A. Steeby, Susan K. Kingsbury and Craig S. Tucker
Department of Aquaculture and Allied Aquacultures
Auburn University
203 Swingle Hall
Auburn, Alabama 36849 USA
trh0008@auburn.edu
A two-year study was conducted to evaluate the second (fingerling to stocker) and third (stocker to growout) phases of a threephase,
modular production system for channel catfish in commercial size farm ponds. Fingerling channel catfish (13.6 g, 12.0
cm) were stocked into each of six earthen ponds (1.62 ha) at a density of 123,500/ha and fed a 35 % crude protein floating feed
daily to satiation. After 142 days, harvested stockers ranged from 109.0 g to 158.9 g (mean = 142.3 g). Survival ranged from
38.8 % to 62.0 %, (mean = 47.7 %), approximately 20 % less than that obtained in small experimental ponds. Individual stocker
fish cost of production was approximately $0.36 each.
Harvested stocker fish were then stocked into 1.62 ha ponds at either 12,350/ha or 16,055/ha for final growout to harvest size
(phase 3). There were three replicates per treatment, and were fed a 35 % crude protein floating feed to satiation with a cap
of 224 kg/ha. Mean total production, combined from an early selective harvest, between 105 to 130 days, and final harvest,
between 207 to 245 days, for the 12,350/ha and 16,055/ha treatments were 7124 kg/ha and 7210 kg/ha respectively. Survival
in growout phase ponds ranged from 84.3 % to 105.5 % (mean = 92.5%) and feed conversion ratios ranged from 2.25 to 2.64
(mean = 2.43) with no significant treatment-dependent effects. Mean weight (0.66 kg) and length (41.86 cm) of fish harvested
from the 12,350/ha treatment was significantly higher than those of fish harvested from the 16,055/ha treatment (0.47 kg; 38.35
cm). Preliminary results from this commercial-scale trial of phase three production are encouraging and the economics of this
final phase will be presented.
Additional advantages of the three phase modular system are its ability to eliminate mortality from bird depredation, to better
control fish inventory, to control harvest size, and to reduce losses from adverse water quality.

SPERM MOTILITY ACTIVATION IN THE ESTUARINE FISH Fundulus grandis
CONDITIONED IN DIFFERENT SALINITIES
Shaunna N. Harris*, Rafael Cuevas-Uribe and Terrence R. Tiersch
Aquaculture Research Station
Louisiana State University Agricultural Center
Baton Rouge, LA 70820 USA
snharris@agcenter.lsu.edu
Sperm motility in externally fertilizing fishes typically responds
to levels of specific ions and the difference in osmotic pressure
between the surrounding water and the body tissues. Usually,
the sperm of marine fish are activated by an increase in osmotic
pressure (hypertonic salinity), and that of freshwater species by a
decrease (hypotonic). These species exist in relatively stable environments.
Estuarine fish, however, are exposed to wide salinity
ranges, often resulting in external osmotic pressures that include
those of the body (isotonic).
To assess the ability of F. grandis sperm to adapt to changes in
salinity, groups of five adult males were acclimated to salinities
of 0, 5, 10, 20, 35, or 50 ppt for one month. Testes were dissected
from the fish and sperm were activated with various osmolalities
of Hanks’ balanced salt solution (HBSS) and calcium-free HBSS
(Ca-F HBSS). The osmolality eliciting the highest motility was
significantly different (P < 0.01) among acclimated groups (Figure
1). It was also evident that calcium or other ions play a role
in motility which peaked after 30 sec exposure to HBSS, and decreased
over 10 min (Figure 2, top). Motility exhibited a similar
initial pattern when exposed to Ca-F HBSS. However, sperm
gained motility at lower osmolalities over 10 min, exhibiting multiple
peaks (Figure 2, bottom). This may aid in explaining apparent
contradictions in research and be of importance in spawning and
cryopreservation of estuarine fishes.
FIGURE 2. Sperm motility after exposure to HBSS
or Ca-F HBSS. Solid lines indicate motility at 30
sec after exposure, dashed lines at 2 min, and dotted
lines at 10 min. These males were acclimated to 35
ppt.
141
FIGURE 1. Osmolality of HBSS (circles) eliciting
highest sperm motility in males acclimated to
various salinities. Points sharing letters were not
significantly different. There was no difference
among blood plasma osmolality (squares) of the
males.

142
EFFECTS OF SMOKING ON THE QUALITY AND SHELF LIFE Hypophthalmichthys molitrix
OF STORED IN 4° c
Sayedeh Fatemeh Hashemi Keneti, Hadi Ershad Lang Rudi and Reza Safari
BSc,Young Researchers Club
Islamic Azad University Branch of Lahijan
PO Box 1616
Lahijan
fh_kenedi@yahoo.com
Nowadays fish and fish products are too valuable and include an important part of white protein to human consumption in the
world. Smoking increase the shelf life of fish. In this research 30 samples with weight average of 850-1000 gr were stored in
4° c tempreture during 120 days. In the present investigation the shelf life of cold smoked Hypophthalmichthys molitrix has
studied and biochemical factors such as protein, lipid and quality factors such as total volatile nitrogen (TVN), proxide value
(PV), mesophilic bacteria and psycrotrophic bacteria were determined during storage.The results showed that the amount of
lipid, protein, proxide value, total volatile nitrogen and total bacteria had significant these results were subjected as anova,
duncan and analysis of variation which were significant at 95% level (p

ENERGY EFFICIENCY AND ENERGY USE REDUCTION IN AQUACULTURE
OPERATIONS AND SUPPORT BUILDINGS
Michael Hatten, P.E.
Solarc Architecture and Engineering, Inc.
Eugene, OR 97401 USA
mikeh@solarc-ae.net
Energy use associated with buildings accounts for 48% of the total energy use in the United State. Many greenhouse gas
reduction initiatives have focused on reducing the energy use requirements of buildings. In the commercial building sector,
this focus has been primarily on opportunities to design and construct new highly efficient buildings that exceed applicable
state and national energy standards. In the industrial sector, the focus has been on upgrade and improvement to existing infrastructure.
Aquaculture operations and support buildings offer opportunities for both efforts – new construction and existing
building system upgrade.
Energy efficiency in new design and construction offers the opportunity to cost-effectively exceed applicable energy standards
by 20% to 50% on the basis of avoided utility costs alone. Consideration of future economic value of avoided emissions is
likely to increase the cost-effectiveness threshold significantly. The State of Oregon’s State Energy Efficient Design (SEED)
program requires all state projects to exceed the energy code by at least 20%. This involves direct involvement with the design
of the project, so that energy efficiency options can be identified for heating, cooling, ventilation, lighting, water heating
and miscellaneous electric loads. Involvement at the early design stages is emphasized. Energy modeling and life cycle cost
analysis is required. The Oregon Hatchery Research Center located in Western Oregon is a good example. The research building
meets the SEED criterion using an integrated set of energy strategies including daylight harvesting, additional insulation,
high performance windows, high efficiency potable water heating, and heat recovery.
Energy use reduction in existing buildings requires additional effort to identify and analyze opportunities. Existing building
systems can be upgraded to newer technology that results in markedly reduced energy consumption often with overall improvement
in system performance. Lighting technology, for example, has advanced to the point where linear fluorescent light
sources perform much better in high bay applications than conventional high intensity discharge (HID) point sources such as
metal halide. Warehouses, storage bays, and covered outdoor canopies that are currently lit with HID can be retrofit with T8 or
T5 linear fluorescent sources and controlled with occupancy sensors. The resulting energy use reduction can be 50% or greater.
The resulting greenhouse gas reductions can also be significant, especially in geographical areas that have a coal-dominated
generation portfolio. Many existing buildings offer energy use reduction opportunities that have little or no capital cost. These
opportunities are associated with operations and maintenance improvements – adjusting temperature setpoints, tuning operating
schedules, optimizing ventilation rates, and fixing broken equipment. Experience with building operations initiatives in the
Pacific Northwest over the past 3 years suggests that buildings can typically reduce energy use by 10 to 15% through straightforward
tune-ups of building energy using systems.
The combination of efficient new construction, cost-effective energy retrofit, and comprehensive building tune-ups offer the
opportunity to reduce energy use in aquaculture operations and support buildings by up to 50%. Harvesting this opportunity
will be an important part of an overall effort to reduce greenhouse gas emissions associated with aquaculture operations nationally
and world-wide.
143

144
EXPLORING THE UTILITY OF A GIS IN EVALUATING THE RISKS AND IMPACTS OF
CATFISH TREMATODE TO ARKANSAS AQUACULTURE
Alf Haukenes*, Larry Dorman and Andrew Goodwin
Aquaculture/Fisheries Center of Excellence
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601 USA
ahaukenes@uaex.edu
Standardized surveys targeting the incidence of specific fish diseases can provide a wealth of information that can be integrated
into a GIS to examine spatial relationships characterizing disease risk. A GIS provides a working format for both the analysis
and the display of complex epidemiological interactions in time and space. Such an approach is useful when identifying the
influence of risk factors or describing potential impacts of fish pathogens as well as distributing ‘real-time’ information to local
extension personnel. We are exploring the use of a GIS to characterize the distribution of catfish trematode in commercial
aquaculture ponds. A two-year sampling effort (2006-2007) was conducted in the state of Arkansas to collect data describing
the prevalence and intensity of catfish trematode infections in commercial ponds. The survey included observations on the
presence of catfish infected with the parasite and on other potential risk factors including the presence of the bird and snail hosts
for the parasite. A simple exploration of the data illustrates the geographic distribution of certain risk factors for the establishment
of the disease (Fig. 1). Catfish farming is a widely distributed in Arkansas and the intermediate snail host for the catfish
trematode was found in ponds from 10 of the 12 sampled counties. Four counties contain ponds where the multiple risk factors
of the snail host and moderate to heavy bird populations were present. In 2006, Ashley and Chicot counties were the only two
counties that contained ponds affected by catfish trematode, however these two counties are the dominant producers of channel
catfish in the state. We are combining the results of the 2007 catfish trematode survey to our GIS and will begin to focus on
different geographic scales to evaluate the distribution of catfish trematode in order to determine factors that help define the
expansion or contraction of the range of distribution of catfish trematode in the state of Arkansas.

CONCENTRATION OF ORGANIC MATERIAL IN OYSTER (Crassostrea corteziensis)
GROWING WATERS AT BOCA DE CAMICHIN, NAYARIT, MEXICO
Maria C. Haws*, Guillermo Rodriguez Domínguez, Omar Calvario Martinez, Laura Edith Corona Osuna
and Eladio Gaxiola Camacho
Pacific Aquaculture and Coastal Resources Center
University of Hawaii at Hilo
Hilo, Hawaii 96720 USA
haws@aol.com
The native oyster species, Crassostrea corteziensis, has been cultured in the Boca de Camichin estuary in Nayarit, Mexico for
over 35 years. Oysters are grown suspended from floating rafts and culture. Recent rapid development without planning or
limitation, has led to concern amongst growers as to the carrying capacity of this heavily utilized body of water. A study to estimate
carrying capacity has begun. The results of water sampling to determine seston concentrations (Total Particulate Matter-
TPM) and the organic fraction of seston (Particulate Organic Matter-POM) are presented here. Water sampling was conducted
between February and June, 2008. Three transects were sampled: at the mouth of the estuary, in the central part of the oyster
growing area, and the interior estuary area. At each of the transects, measurements were taken at 1.2 m and 2.5 m depth from
the center of the estuary channel and close to the two banks. Sampling was repeated at ebb and flood tides.
The concentration of TPM varied from 9.2 μg/ml in February to 109 μg/ml in June with a mean of 39.18 μg/ml. The PMO
varied between 2.06 μg/ml in February to 6.59 μg/ml in March with a mean of 3.82 μg/ml. There was no significant difference
in concentrations of TPM nor PMO during the two tidal stages (Table 1).
These concentrations are higher than found at another oyster (Crassostrea gigas) growing area in Bahia San Quintin in Baja
California, Mexico (García-Esquivel, 2004) where the average POM concentration was 1.9 μg/ml in an area where 1,913 tons
of oysters are produced annually. POM as a percentage of seston (TPM) was lower however, in Boca de Camichin (3.1 to
22.4%) than at San Quintin (17.8 to 25.5%). The higher proportion of POM may influence the oysters’ assimilation rate.
14

14
DEVELOPMENT OF A FLEXIBLE PANEL FISH GRADING SYSTEM
David Heikes
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601 USA
dheikes@uaex.edu
A flexible panel grading system has been designed and fabricated. The grading panels consist of short lengths (10.25”) of
sunlight resistant rigid pvc conduit (1/2” schedule 40) strung on galvanized aircraft cable (3/16”, 7x19 construction) with industrial
plastic spacers. The individual panel dimensions are approximately 4’ by 20’. The individual apertures can be produced
in 1/8” increments (ie. 1.84”, 1.965”, 2.09”, etc) through various combinations of commercially available industrial spacers.
Three flexible grading panels (2.09” aperture) were fabricated and sewn into the sidewall of a standard–built harvesting sock
60’ long with 1” mesh. Preliminary tests indicate that this design allows hybrid catfish to passively grade without any other
major change to harvesting operations. Farm workers can handle and crowd fish from the modified sock in a similar fashion
to standard socks.
Additionally, work has begun to determine the optimal aperture for grading hybrid catfish with the flexible grading apparatus.
Three small test panels (4’ x 5’) were fabricated with 1.84”, 2.09”, and 2.34” apertures. These panels were incorporated into
three small socks designed to hold 2000 lbs of catfish each. Figure 1 shows the number of hybrid catfish retained by the various
bar apertures as compared to the population prior to grading. Fish were allowed to passively grade overnight (approx.
12 hours). This grading system makes passive sock grading a realistic option for harvesting food-size channel x blue catfish
hybrids and possibly other commercially produced species.

VHS BIOSECURITY WORKSHOPS AND THE DEVELOPMENT OF FISH DISEASE HACCP
PLANS IN THE NORTH CENTRAL REGION OF THE US
Jeffrey A. Malison*, Ronald E. Kinnunen, Ronald E. Johnson, Sarah E. Kaatz and James A. Held
University of Wisconsin-Stevens Point Northern Aquaculture
Demonstration Facility (NADF)
College of Letters and Science
Stevens Point, WI 54481 USA
edjayson@seafdec.org.ph
In September, 2008 we will begin a significant extension and outreach project focusing on VHS and other fish diseases. The
project will be funded by the North Central Regional Aquaculture Center (NCRAC). We plan to conduct a series of six biosecurity
workshops held at different fish farms across the region, targeting flow-through, pond, and recirculation systems. We will
utilize the existing Aquatic Invasive Species Hazard Analysis Critical Control Point (HACCP) Training Curriculum to develop
specific fish disease HACCP plans for each of the six facilities involved in the workshops. The workshops will utilize extension
materials developed by the Iowa State University Center for Food Security and Public Health (CFSPH), including a VHS
response package and PowerPoint presentation. The workshops will be organized in close collaboration with local fish farmers
and appropriate state aquaculture associations. Each workshop will include presentations and demonstrations delivered by
individuals with diverse areas of expertise in aquaculture. These will include NADF and other regional aquaculture extension
specialists, private fish farmers, representatives from state aquaculture associations, regional aquatic veterinarians or fish health
specialists, state regulators, and national regulatory, research, and advisory agencies.
We will then develop and distribute three model fish disease HACCP plans, one each for flow-through, pond, and recirculation
systems, relying heavily on the specific plans developed. The model HACCP plans will be made available electronically
through the NADF, CFSPH, and NCRAC Web sites, and by hard copy.
We will also capture all of the proceedings of the workshops and HACCP development efforts on video. Using this footage, a
cohesive video will be prepared that will detail all of the key issues involving VHS, fish diseases in general, and fish disease
HACCP plan development. This video will be made available on DVD as well as streaming video on the aforementioned Web
sites.
14

14
DOUBLE CROPPING OF YELLOW PERCH Perca flavescens FINGERLINGS IN PONDS
James A. Held*, Jeffrey A. Malison, Amanda J. Smith and Christopher Hartleb
University of Wisconsin-Extension
Northern Aquaculture Demonstration Facility
302 S. Main St.
Lake Mills, WI 53551 USA
jaheld@wisc.edu
One factor limiting the expansion of commercial yellow perch production is the availability of fingerlings. Yellow perch are
group synchronous spawners, with each female producing a single ribbon of eggs in the spring. These eggs give rise to a single
cohort of fingerlings produced in fertilized ponds. When the fry reach 18-25mm total length (TL) and approximately 6 weeks
old, they are removed from ponds and habituated to formulated feed. Once trained, the fingerlings are returned to ponds or
placed in recirculation systems for grow-out. The production of a second crop of eggs each year would enhance the profitability
of fingerling production and potentially double the number of fingerlings available for grow-out.
Out-of-season spawning of perch, where the broodstock are maintained on phase-shifted temperatures and photoperiods, has
proven expensive, unreliable and logistically difficult. In 2008 we employed a protocol that combined early spawning of the
broodstock with delayed spawning and protracted egg incubation to attain double cropping of fingerling production ponds.
Broodstock were removed from ponds shortly after ice out (April 1). Females for the first spawn were injected with human
chorionic gonadotropin (hCG, 50 I.U./fish) and held in tanks until eggs were stripped (6-8 days after injection). Females for the
second spawn were held in a tank on cold (5-7°C) water and short (8 hr) day lengths. Eggs from the first spawn were incubated
at 15°C and hatched 8 days later. Hatched fry were stocked into ponds on April 16. Fingerlings were harvested from ponds on
May 29 at approximately 20 mm TL (0.25g) and stocked into tanks for habituation to formulated feed. The females being held
for the second production cycle began to spawn spontaneously in the tank on May 8. Fish were spawned and ribbons collected
without the use of hCG on May 12-15. Egg ribbons from the second spawn were incubated at either 3°C or 5°C and hatched
in 27 and 20 days, respectively. Hatched fry were stocked into previously depopulated and refilled fingerling production ponds
(June 4-8). Fingerlings were harvested from ponds on July 14-15 at 30-50mm TL (1.0-2.5g) and transferred to tanks for feed
training.
Observation of embryonic development during incubation revealed differences between the three incubation regimens. Normal
development was noted in eggs incubated at 15°C, normal but slowed development was noted at 5°C, and eggs incubated at 3°C
displayed a high percentage of developmental abnormalities that became apparent in the final stages of embryonic maturation.
The most obvious of these abnormalities was scoliosis of the caudal region. Further study is needed to determine if temperature
manipulation during critical developmental stages can reduce or eliminate these abnormalities. Throughout the course of the
two pond production phases, we compared the succession of phytoplankton, zooplankton, and benthic organisms.
This study was supported by grants from the USDA-SBIR program and the Wisconsin Sea Grant Institute.

PROSPECTS FOR ENGINEERING SOYBEANS OPTIMIZED AS FISHMEAL REPLACERS
IN AQUACULTURE FEED
Eliot Herman*, Theodore Hymowitz, Wayne Parrot and Monica Schmidt
USDA/ARS
St. Louis MO 63132 USA
eherman@danforthcenter.org
Soybean protein replacement challenge feeding tests have shown that a relatively low percentage of soybean in the diets of
Salmonids results in adverse growth. Resolving this problem will make soybean a desirable renewable protein source to support
the growing aquaculture industry. Engineering soybean by either germplasm selection and breeding or by directed genetic
modification offers the prospect of altering multiple traits in soybean not only eliminating the potentially growth limiting proteins
and compounds but also to enhance or introduce new traits to create a soybean optimized for Salmonid growth.
Current research in our laboratories is directed at stacking potentially useful traits in soybean including nulls for major allergens
and anti-nutritional proteins and to express carotenoids at high concentrations. Conventional nulls for the immunodominant
human allergen P34 as well as lectin and Kunitz trypsin inhibitor have been stacked producing a triple null for these three
proteins. As a control a genetically modified soybean has been created that hyper-accumulates the same three proteins. The
anti-nutritional and allergenic components in soybean have been implicated in the feed intolerance of Salmonids. By creating
soybeans null for these proteins this may improve the inclusion levels permissible in Salmonid feed. Both the triple null and the
hyper-accumulating soybean have been expanded in the field for feed challenge tests on Salmonids during 2009.
To investigate the potential for adding high-value nutrients to soybean β-carotene has been accumulated in soybean as a result
of genetic modification. The resulting soybeans accumulate much higher levels of β-carotene than other examples such as
Golden Rice. The β-carotene is precursor to produce other carotenoids including astaxathin. By accumulating a larger precursor
pool this is directed at driving the carotenoid pathway toward accumulating the pigment needed for Atlantic Salmon and Char
production stacked with other traits such as anti-nutritional/allergen nulls.
By creating soybeans with multiple improved traits and by combining germplasm selection, breeding with genetic modification
it is feasible to create specialty value-added lines of soybean directed for use by specific end-users such as aquaculture.
Soybeans specifically optimized for aquaculture will provide a renewable consistently available protein source for the growing
Salmonid world-wide production. This work will contribute to the NOAA-USDA Plant Products in Aquaculture Working
Group’s Strategic Research Plan Goal 3: “Enhance the inherent composition of crops to provide a beneficial balance of bioactive
compounds in order to optimize their use in aquafeeds for carnivorous fish.”
14

1 0
EVALUATION OF SUBSTRATES IN BIOLOGICAL FILTERS FOR REARING PACIFIC
WHITE SHRIMP Litopenaeus vannamei IN CLOSED SYSTEMS
Mario Hernández-Acosta, Abundio González-González, Gilberto Gutiérrez-Salazar, Francisco M Guzmán-Saenz,
Jorge Loredo-Osti, Pablo González-Alanis and Kevin Fitzsimmons
Environmental Research laboratory
University of Arizona
2601 E. Airport Dr.
Tucson, Az 85706 USA
mariohdz@email.arizona.edu
The use of Biofilters to control water quality for long periods, offers the possibility of commercially producing marine organisms
at inland sites, avoiding problems such as environmental contamination. Recirculating water systems require the use
of a denitrification process to keep survival and ammonium and nitrate levels within acceptable ranges in order to insure the
cultivated marine organism’s development. Biofilters are considered as the most important factor for water treatment in a
recirculating system; their purpose is to provide a substrate to let probiotic bacteria grow. The development of such biofilters
has focused on effectiveness and conserving a proper balance between ammonia production and removal. Six different biofiltration
treatments were used in this study. The two way analysis of variance shows significant differences in survival of shrimp.
The physical – chemical parameters such as: pH, N-NO 2 y N-NO 3 , were similar in all treatments (p > 0.05); but the DO, TAN,
N-NH 3 and CaCO 3 were statistically different (p < 0.05). The two best biofilter systems with regard to shrimp survival and
growth, present an opportunity to culture shrimp for long periods with cero water exchange
.
Treatments
1 2 3 4 5 6 p
Temp ( 0 C) 26.7 ± 0.66 26.7 ± 0.66 26.7 ± 0.66 26.7 ± 0.66 26.7 ± 0.66 26.7 ± 0.66 p > 0.05
DO (mg/L) 5.08 ± 0.31 5.05 ± 0.54 4.97 ± 0.37 4.75 ± 0.33 4.40 ± 0.49 4.50 ± 0.37 p < 0.05
pH 7.70 ± 0.11 7.70 ± 0.11 7.70 ± 0.11 7.72 ± 0.10 7.62 ± 0.16 7.68 ± 0.13 p > 0.05
TAN (mg/L) 0.125 ± 0.027 0.122 ± 0.045 0.100 ± 0.077 0.142 ± 0.097 0.063 ± 0.022 0.075 ± 0.042 p < 0.05
N-NH3 (mg/L) 0.0038 ±0.0017 0.0033 ±0.0015 0.0028 ±0.0022 0.0042 ±0.0029 0.0015 ±0.0006 0.0018 ±0.0012 p < 0.05
N-NO2 (mg/L) 0.375 ± 0.154 0.492 ± 0.246 0.300 ± 0.089 0.467 ± 0.266 0.292 ± 0.156 0.283 ± 0.264 p > 0.05
N-NO3 (mg/L) 7.3 ± 2.3 8.2 ± 2.2 7.7 ± 2.7 8.7 ± 2.1 8.8 ± 2.0 9.0 ± 2.4 p > 0.05
CaCO3 (mg/L) 248.7 ± 12.6 236.7 ± 26.3 219.0 ± 18.7 221.3 ± 19.5 192.7 ± 37.2 198.3 ± 31.4 p < 0.05
Salinity (ppt) 15 ± 0 15 ± 0 15 ± 0 15 ± 0 15 ± 0 15 ± 0 p > 0.05
Table 1.- Mean and standar deviation of the evaluated parameters during the experiment.

ROE AND CAVIAR BACTERIOLOGY: In vivo, In situ, In sushi
Brian Himelbloom* and Chuck Crapo
University of Alaska Fairbanks
School of Fisheries and Ocean Sciences
Fishery Industrial Technology Center
118 Trident Way
Kodiak, AK 99615-7401 USA
ffbhh@uaf.edu
Fish roe and caviar are delicacies consumed worldwide. These raw eggs are marketed as salt-cured whole skeins (sujiko) and
singles (ikura). Bacteria may be present in either product since no processing treatment is used for inactivation or prior to
consumption. Concerns have been raised regarding the potential for pathogenic bacterial species being present in these products.
Also, the shelf life of the product depends on the conditions at wholesale and retail and handling by the end user at the
restaurant, supermarket or home. This review covers the aspects of roe and caviar bacteriology regarding live fish in vivo, egg
deposition in nature in situ and as products for consumers in sushi.
The literature is scant for the first two instances in which fish eggs are contained in the female fish or released during spawning
and obtained for aquaculture fertilization. Unless the fish is diseased, the ovaries are considered bacteriologically sterile.
Exposure to the environment in streams or hatchery tanks does allow bacteria to become associated with the eggs upon release
from the female. In either situation, damage to egg integrity can provide entrance by fish pathogens to the live eggs. Otherwise,
nature has provided the eggs a protective barrier to bacteria invasion but not necessarily against bacterial adhesion.
Handling and processing into whole skeins or caviar are operations that the fish processing facilities isolate from other activities.
These highly valuable commodities command high prices due to species preference, limited availability and intrinsic
factors such as egg size, color and shape. Caviar grades, as decided by corporate inspectors and buyer specifications, are
based on the raw characteristics for batches of sujiko and ikura. Bacteriological content and species identification are analyzed
separately to evaluate the safety and quality of these products. If declared unsafe by regulatory agencies, sujiko and ikura are
recalled from the market.
1 1

1 2
THE LAPAZ PROJECT: AN INDUSTRY-UNIVERSITY ENTERPRISE FOR PRODUCTION
OF ATLANTIC STURGEON
Jeffrey M. Hinshaw and Thomas M. Losordo
Department of Biology
North Carolina State University
455 Research Drive
Mills River, NC 28759 USA
Large scale commercial culture of sturgeon in the United States is limited to a small number of enterprises in Florida, California,
and Idaho. Facilities in Florida are producing primarily Siberian sturgeon Acipenser baerii, while the facilities in
California and Idaho are limited to growing white sturgeon A. transmontanus. Private facilities in other locations in the U.S.
(e.g., Hawaii,) are growing a limited number of Russian sturgeon A. gueldenstaedtii. To date, culture of Atlantic sturgeon A.
oxyrinchus has occurred primarily at public hatcheries or university research facilities. In 2004, faculty at North Carolina State
University began working with a commercial partnership in Lenoir, NC, to plan development of facilities for Atlantic sturgeon
culture for caviar and meat production. The ‘LaPaz’ project has now evolved into a research and demonstration facility for the
commercial production of sturgeon, primarily Atlantic sturgeon, but also including a limited number of Russian sturgeon and
Siberian sturgeon.

A ZEBRAFISH MODEL TO INVESTIGATE THE PATHOGENICITY AND PERSISTENCE OF
VIRAL HEMORRHAGIC SEPTICEMIA VIRUS IVB
Kristine M. Hope*, Paul R. Bowser and James W. Casey
Cornell University
Department of Microbiology and Immunology
C4-134 Veterinary Medical Center
Ithaca, NY 14853-6401 USA
kmh58@cornell.edu
Viral hemorrhagic septicemia (VHS) is considered the most important viral disease of finfish worldwide (Wolf 1988) and is
listed as a reportable disease by national and international organizations. In 2005, reports of massive die-offs from a related
strain of VHSV in the Great Lakes (VHSV-GL), now identified as genotype IVb (Elsayed 2006), conclusively signaled an early
invasion of VHSV. To more precisely monitor VHSV-GL we have used quantitative RT-PCR to measure the prevalence of
virus and replicative intermediates in fish tissues and as free virus in water. We have applied these nucleic acid tools and initiated
Zebrafish infection studies to investigate the in vivo infectious cycle and pathogenesis of VHSV-GL. The Cornell strain
of Zebrafish (Danio rerio) was bath challenged with varying viral titers, age groups, and different times of adaptation at 15°C.
This low temperature adaptation is required since viral replication only occurs at low temperatures. The host immune system
is also thought to be immunosuppressed at this temperature. Moribund fish displayed clinical signs, which included petechial
hemorrhage at the base of the fins and abdomen, distended visceral cavity, exophthalmia, and erratic swimming behavior. Microscopic
evaluation revealed focal hemorrhage in the dermis and muscle. Both moribund and apparently normal VHSV-GL
exposed Zebrafish showed extensive variability in viral loads that likely represent time of sampling or suggest that some fish
may be more resistant to viral infection due to individual variability within the population and/or immune response mechanisms
that temper the pathogenic outcome. Mortality primarily depended on infectious dose. At high levels (10 6 /ml) an initial mortality
of 20% was observed at day 13 followed by a second wave starting at day 23 and reaching a total of 80% mortality at day
33. This second phase of mortality correlates well with viral shedding from initially infected zebrafish since water samples
from day 12 contained an average of 10 4 viral genomes per ml. Unexpectedly, infection at 15°C after a 0 hour adaptation period
resulted in 100% mortality in 6 days. This surprising latter result suggests that temperature shock, as occurs frequently in the
Great Lakes, may be a key abiotic factor that results in mass mortality and may more precisely define species susceptibility.
Comparative pathogenesis of VHSV-GL with the less virulent marine isolates of VHSV, in terms of innate immune response,
replication levels and shedding may also provide insights into pathogenic traits.
1 3

1 4
HISTORY AND IMPROVEMENTS OF CRYOPRESERVATION OF PADDLEFISH SPERM
Ákos Horváth*, Béla Urbányi and Steven D. Mims
Department of Fish Culture
Szent István University
Godollo H-2103, Hungary
Horvath.Akos@mkk.szie.hu
Paddlefish belongs to the order Acipenseriformes and it is closely related to sturgeons, therefore, the biology of paddlefish
sperm is also similar to that of sturgeons. This is characterized by a lower osmolality of the seminal plasma (70-100 mOsmol
kg -1 compared to 300 mOsmol kg -1 in teleosts), the presence of a functioning acrosome on the tip of the spermatozoon head and
numerous micropyles on the surface of eggs. Accordingly, problems associated with cryopreservation of sperm were similar,
as well. Sturgeon and paddlefish sperm cryopreservation was considered very problematic for several decades. Published
protocols shared a common problem: although high post-thaw motility of sperm was observed, fertilization and hatch percentages
remained low and were inconsistent. In general, the cryoprotectant dimethyl-sulfoxide (DMSO) was employed and little
attention has been paid to the osmolality of the extender apart from verifying that it keeps spermatozoa immotile.
In the late 1990-es protocols have been published on the cryopreservation of sperm from several sturgeon species, including
the Siberian sturgeon Acipenser baeri and the sterlet A. ruthenus with satisfactory fertilization and hatch rates. In these experiments
methanol was used as a cryoprotectant and the osmolality of the extenders was (albeit unintentionally) kept at around
70-80 mOsmol kg -1 . Following several modifications of the cryopreservation protocol it was applied to the sperm of paddlefish
and satisfactory fertilization and hatch results were observed. When sperm was frozen in 0.5-ml French straws and in the presence
of 10% methanol a fertilization percentage of 80 ± 3 % was observed (in contrast to 17 ± 10 % using DMSO). Using the
same sample a hatch rate of 79 ± 5 % was achieved later. To increase the volume of sperm frozen in a single unit (straw) and to
allow the fertilization of larger batches of eggs with the contents of a single straw, sperm was frozen in 4- and 5-ml macrotubes.
Hatch percentages of up to 77 ± 7 % were observed using these straws.
The protocol that is currently used by our group is as follows: sperm is diluted at a ratio of 1:1 in Modified Tsvetkova’s extender
(23.4 mM sucrose, 0.25 mM KCl, 30 mM Tris, pH 8.0) to which 5 or 10 % methanol is added. Diluted sperm is loaded into
0.5- or 5-ml straws and placed on a polystyrene frame that floats on the surface of liquid nitrogen in a polystyrene box. The
distance from the surface of nitrogen is 3 cm, the cooling time is 3 min. Straws are thawed at 40 °C for 13 sec (0.5-ml straws)
or 40 sec (5-ml straws).

SIZE AT SEXUAL MATURITY AND FECUNDITY INDICES OF THE INDIAN RIVER
SHAD Gudusia chapra (CLUPEIDAE) IN THE GANGES RIVER OF THE NORTHWESTERN
BANGLADESH
Md. Yeamin Hossain and Zoarder Faruque Ahmed
United Graduate School of Agricultural Sciences
Kagoshima University
1-21-24 Korimoto
Kagoshima 890-0065
Japan
yeamin2222@yahoo.com
The Indian river-shad Gudusia chapra (Hamilton-Buchanan, 1822) is a small indigenous fish species of Bangladesh and widely
distributed in the natural waters of Asia, throughout the Indian subcontinent including Bangladesh, India, Pakistan, Nepal, Bhutan,
Sri-Lanka, Myanmar, Afghanistan. This small indigenous species (SIS) used to be abundantly available in rivers, streams,
ponds, beels, ditches, and floodplains in the past in the South Asian region, but nowadays it is disappearing from the natural
systems, which in turn severely affects biodiversity. The recent socio-economic studies identified that G. chapra as both an
important food resource and a crucial source of micronutrients essential in preventing malnutrition and vitamin and mineral
deficiencies in rural communities, particularly among women and children in Bangladesh and making it a highly favorable
candidate for aquaculture in the South Asian countries.
The present study aimed to estimate the size at sexual maturity and fecundity for female G. chapra from the Ganges River of
northwest of Bangladesh during March to August 2006. The gonadosomatic index (GSI) was calculated by the equation, GSI
(%) = (GW/BW) × 100. The size at first sexual maturity of female of this species was estimated by the relationship between
gonadosomatic index and its standard length. The specimens larger than first size at sexual maturity were used for the estimation
of fecundity.
The results showed that SL of G. chapra ranged from 3.60 to 13.70 cm, with mean ± SD calculated as 8.80±2.93 cm while
body weights varied from 1.00 to 43.60 g with calculated 16.56 ± 11.96 g. The size at first sexual maturity and median sexual
maturity for females G. chapra were considered to be 8.00 cm and 9.80 cm SL, respectively in the Ganges River. The mean
total fecundity was 20160±6545 and ranged from 10888 to 36200. This study would be useful for fishery biologists/managers
to impose adequate regulations for sustainable fishery management in the Ganges River and nearby areas of Bangladesh.
1

1
RAPID TEST FOR THE DETECTION OF NECROTIZING HEPATOPANCREATITIS
BACTERIUM IN TISSUE EXTRACTS AND FECES OF PENAEID SHRIMP
Raymond L. Houghton*, Carlos Pantoja, Bonnie T. Poulos, Jean Chen, Yvonne Y. Stevens, Stan Morkowski,
Syamal Raychaudhuri and Donald V. Lightner
InBios International Inc.
562 First Avenue South, Suite 600
Seattle, WA 98104 USA
Raymond@inbios.com
A rapid lateral flow dipstick assay was developed for detection of Necrotizing Hepatopancreatitis Bacterium (NHP-B) in
penaeid shrimp. The assay is designed to detect the bacterium in extracts of the shrimp hepatopancreas tissue as well as in feces.
Two monoclonal antibodies were used in assay development that were shown by western blot to detect a 13kDa component in
the bacterium. Experimentally infected Litopenaeus vannamei shrimp were harvested and hepatopancreas and fecal samples
collected and extracted. Dipstick reactivity was observed in both sample types when infection was detected by PCR or immunohistochemistry.
The dipstick assay also reacted with Percoll fractions of hepatopancreas enriched for flagellar components of
the organism as shown by electron microscopy.
The rickettsial like organism resulting in Necrotizing
Hepatopancreatitis (NHP-B) in shrimp has resulted in
the loss of millions of dollars in the shrimp aquaculture
business particularly in the Americas. This raises
the need for a rapid test procedure for application by
farmers on site to detect the infection earlier and also
to monitor spread of the disease. Such a test would
need to be able to detect the organism in either fecal or
hepatopancreas samples (HPs) with sufficient sensitivity
to enable a more rapid response time to containing
infections. The availability of a cost effective test to
routinely monitor shrimp cultures in the field as well as
brood stocks could play an important role in identifying
and containing infections early. Two antibodies designated
3D6 and 4A2 have been used in combination in a
lateral flow immunoassay to detect NHP-B in both PCR
positive (4 strong, 2 weak, 17 negative) hepatopancreas
tissue Table 1 as well as feces of penaeid shrimp Figure
1. It has also been used to detect activity in Percoll fractionated
extracts of HPs enriched for flagellar antigens
(results not shown). This test will facilitate monitoring
of NHP-B infection in the field.

EFFECT OF HERBAL LEAF DECOCTION ON PATHOGENICITY OF SOME AQUATIC
SPECIES IN ORGANIC AQUACULTURE SYSTEM
Dr. Hnin Hnin Htun* and Dr. San San Yu
Biotechnology Department
Technological University(Maubin)
Ayeyarweddy Division
Myanmar
In this study showed that the use of herbal leaf decoction in aquaculture system can be an effective method to prevent disease
outbreaks caused by pathogenic bacteria in hatchery and farming. It was used at the various concentration of herbal leaf decoction
in larval rearing, adult of giant freshwater prawns (Macrobrachium rosenbergii) and fingerling stage of rohu (Labeo
rohita) applying modified static to organic farming system. Black spot disease and Aeromonas hydrophila were isolated from
prawn and fish samples. The toxicity test and pathogencity test were conducted. This study was done by three experiments.
Each experiment was conducted by three different concentrations of herbal leaf decoction in trails with 2-replicated of each as
(i) control (ii) 10 ml/ton (iii) 20 ml/ton (iv) 30 ml/ton were set up. In the experiment I, trail 3 is the best among trails. The first
PL was found in this trial at day (35) th , the total bacterial count was the lowest and PL were less sensitive to pathogenic bacteria.
In the experiment II, trail 3 is the best among trails. The first recover adult prawn was found in this trail at 1 to 2 weeks.In the
experiment III, trail 3 is the best among trails. The first recover fingerling fish was found in this trail at 5 to 7 days.Therefore,
the possible use of herbal leaf decoction in prawn and fish farming, we evaluated the in vitro and in vivo experiments herbal
leaf, neem decoction against the infection by pathogenic bacteria (shell-degrading bacteria).
1

1
Table 1. Comparison of two straw types used for cryopreservation
HIGH-THROUGHPUT CRYOPRESERVATION Features IN AQUATIC CBS SPECIES: ADAPTING
MAMMALIAN TECHNOLOGY TO FISH
E Hu*, Huiping Yang and Terrence R. Tiersch
Aquaculture Research Station
Louisiana State University Agricultural Center
Baton Rouge, LA 70820 USA
ehu@agcenter.lsu.edu
TM straws French straws
Physical
high elasticity low elasticity
characteristics
identification jacket
transparent (colored
no external cover
transparent or colo
Economics
$0.51 per straw $0.04 per straw
sold in packages of 300 sold in packages o
Features of products heat sealing of both ends
0.3-ml or 0.5-ml volume
gel plugging one e
0.25-ml or 0.5-ml v
With the adoption of existing commercial technology, high-throughput
cryopreservation is possible in aquatic species. As a first step, our goal
was to adapt an automated system developed for livestock and humans
for use in channel catfish (Ictalurus punctatus). Our objectives were to:
1) compare existing and high-throughput fish sperm cryopreservation
procedures; 2) compare physical characteristics of Cryo Bio System
(CBS TM ) straws and standard French straws, and 3) begin initial research
on fish sperm for high-throughput cryopreservation.
For straw filling, sealing, and labeling, we used the CBS TM MAPI system
(www.cryobiosystem-imv.com) to provide high biosecurity and
large-scale production. There are a number of differences between the
straw types (Table 1), and the thermal properties of the straws were studied
(Figure 1). Using standard conditions, the CBS straws cooled more
slowly and required a longer thawing time. Based on our preliminary
studies for channel catfish, the cryoprotectant selected was 5% methanol,
the cooling rate was 20 ºC/min, and the thawing conditions were 40
ºC for 23 sec. More technical and economic research are necessary to
adapt the MAPI system for aquatic sperm, but it offers great potential for
culture and conservation.
Table 1. Comparison of two straw types used for cryopreservation
Features CBS TM straws French straws
Physical
high elasticity low elasticity
identification jacket no external cover
characteristics transparent (colored transparent or colored
Economics
$0.51 per straw $0.04 per straw
sold in packages of 300 sold in packages of 2000
Features of products heat sealing of both ends gel plugging one end; other end
0.3-ml or 0.5-ml volume 0.25-ml or 0.5-ml volume
Figure 1. Temperature profiles of the 0.5ml
CBS straw (dotted line) and the 0.5-ml
French straw (dashed) in response to
cooling at 40 ºC/min (top) and thawing at
40 ºC (bottom). The solid line is the
freezing chamber temperature. The
horizontal axes present time (sec); the
Figure 1. Temperature profiles of the 0.5ml
CBS straw (dotted line) and the 0.5-ml
French straw (dashed) in response to
cooling at 40 ºC/min (top) and thawing at
40 ºC (bottom). The solid line is the
freezing chamber temperature. The
horizontal axes present time (sec); the
vertical axis presents temperature (ºC).

AQAUFISH CRSP: FOSTERING THE DEVELOPMENT AND EXCHANGE OF
SUSTAINABLE AQUACULTURE AND FISHERIES MANAGEMENT PRACTICES
WORLDWIDE
Stephanie T. Ichien*, Ford Evans and Hillary S. Egna
AquaFish CRSP
Oregon State University
Corvallis, OR 97333 USA
aquafish@oregonstate.edu
In 2006 Oregon State University entered into a cooperative agreement with the United States Agency for International Development
(USAID) to promote sustainable aquaculture and fisheries worldwide through the Aquaculture & Fisheries Collaborative
Research Support Program (AquaFish CRSP).
The mission of the AquaFish CRSP is to enrich livelihoods and promote health through international multidisciplinary partnerships
that advance science, research, education, and outreach in aquatic resources. Bringing together resources from US and
Host Country institutions, the AquaFish CRSP emphasizes sustainable solutions in aquaculture and fisheries for improving
health, building wealth, conserving natural environments for future generations, and strengthening poorer countries’ ability to
self-govern. The goals of AquaFish CRSP include:
• Increase Host Country productivity and provide return benefits to the US.
• Use innovative research to understand and remove constraints in fisheries and aquaculture facing poorer countries.
• Build US and Host country capacity in aquaculture and fisheries management to ensure long-term impacts.
• Foster wide dissemination of research results and technologies.
• Provide outreach in aquaculture and fisheries topics through publications, short courses and region specific Podcasts.
These goals are accomplished through an integrated, multidisciplinary, cross-cutting research and outreach program that increases
aquaculture productivity, enhances environmental stewardship, addresses gender integration, prevents further degradation
of aquatic ecosystems, and increases domestic and export market opportunities.
The AquaFish CRSP currently manages six research projects that contain 38 investigations. These research projects represent
all of the key regions, themes, and topic areas called for in the request for proposals. As a group they include 14 countries, 12
US universities, and more than 20 Host Country institutions in formal funded partnerships, plus more than 70 additional collaborators
in informal partnerships.
The overall AquaFish CRSP is managed in a manner to achieve maximum program impacts, particularly for small-scale farmers
and fishers, in Host Countries and more broadly. Program objectives address the need for world-class research, capacity
building, and information dissemination.
1

1 0
FEEDING TRIALS TO IMPROVE SURVIVAL AND GROWTH OF PALLID STURGEON
Scaphirhynchus albus LARVAE
Jason Ilgen, Matt Toner and Kevin M. Kappenman
U.S. Fish and Wildlife Service
Bozeman Fish Technology Center
4050 Bridger Canyon Road
Bozeman, MT 59715 USA
jason_ilgen@fws.gov
The Pallid Sturgeon Recovery Program determined captive propagation of endangered pallid sturgeon as necessary to recover
the species and state and federal hatcheries are operating to achieve production goals. Hatchery mangers identified initiation
of larvae pallid sturgeon to feed as the most critical rearing period and stated that mortality during this period can determine
hatchery year class strength. The discontinuation of a diet historically used at many hatcheries and the variety of diets used at
other hatcheries exposed a need for information quantified through analytical research on the effects of different commercial,
experimental, and live feeds on survival and growth of larvae pallid sturgeon. We tested the effects of 5 diets (Corey High
Pro and freeze dried cyclopeeze (FDC), Otohime (OTO), Bio Vita, Bio Vita and frozen cyclopeeze, Bio Vita and FDC) on
larvae survival and growth over 60 d in 2007, and 5 diets (OTO, OTO and FDC, OTO and freeze dried Artemia, OTO and
live Artemia, and ARS experimental diet) for 30 d in 2008. In 2007, we determined a diet of OTO produced relatively high
survival (54%) compared to other stand alone diets tested, and found that supplementing Bio Vita with freeze dried cyclopeeze
improved survival (63%) compared to Bio Vita alone (14%). In 2008, we determined that OTO supplemented with freeze dried
cyclopeeze increased survival (92%) compared to OTO alone (72%). Information from this research can improve efficiency
of captive rearing programs by increasing survival, growth, and fish condition and promote recovery of the species.
Figure shows survival results of five test diets in the 2008 trials. Letters indicate significant differences.
a
b
ab
a
ab

STUDY ON POSSIBILITY OF PRODUCTION OF QUALITY HALF-PEARLS FROM THE
BLACK-LIP PEARL OYSTER Pinctada margaritifera Linnaeus, 1758 IN TANZANIA
Edson Ishengoma*, Aviti J. Mmochi,m Narriman S. Jiddawi and Rashid A. Tamatamah
Edson Ishengoma
Institute of Marine Science
University of Dar-Es-Salaam
P.O.Box 668
Zanzibar
ebarongo82@yahoo.co.uk
We evaluated the possibility of producing half-pearls, using black-lip oysters, P. margartifera, on the Northern Eastern coast of
Tanzania. Two groups of 50 oysters, small and large varieties, both collected from wild, with mean (±S.D.) dorso-ventral shell
height (DVM) of 65.0±6.0 mm and 105.0±26.5 mm, respectively, were implanted with nuclei. In small oysters, two plastic
nuclei were glued one on each valve; and in large oysters three were glued two on one valve and one on the other, were cultured
for a period of ten months. Along with this, mortality, rate of nacre production and thickness of nacre deposited were also
studied. The mortality rates were generally higher for small oysters than for large oysters. For the first one month of culture,
mortality in small and large oysters was recorded at 16% and 42% respectively (Fig 1). The nacre deposition rate was found to
be significantly slower (P >0.05) in small oysters than in large oysters.
In total, 30 half pearls were harvested in small oysters and 51 half pearls harvested in large oysters. In small oysters, no Agrade
half-pearls were obtained, 30% B-grade, 16.67% C-grade and 56.33% deformed. In large oysters, 15.69% were A-grade,
58.82% B-grade, 19.6 C-grade and 5.88% deformed (Fig 2). Quality pearls of good color were mostly produced on the lip area
of the shell on large oysters. The study showed that it was possible to obtain relatively thick nacre within a short period of 5
months in large oysters, deposition rate being about 2 times higher than that observed in small oysters. Furthermore, the study
suggests that small oysters could produce quality half pearls if smaller nuclei are used.
1 1

1 2
DEVELOPMENT OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION COMBINED
WITH A LATERAL FLOW DIPSTICK (LAMP-LFD) FOR DETECTION OF WHITE SPOT
SYNDROME VIRUS
Wansadaj Jaroenram*, Wansika Kiatpathomchai and Timothy William Flegel
CENTEX Shrimp, Faculty of Science
Mahidol University
Rama 6 Road
Bangkok 10400, Thailand
Kungbtram@gmail.com
Loop-mediated isothermal amplification (LAMP) allows rapid amplification of nucleic acid under isothermal conditions using
a set of six specially designed primers that recognize eight distinct target sequences with high specificity and sensitivity.
In this report, a 20-minute LAMP method for amplification of WSSV DNA using biotin-labeled primer was combined with
a chromatographic lateral flow dipstick (LFD) for rapid and simple visual detection of WSSV-specific amplicons. The LFD
process involved a 5 min post LAMP step for specific hybridization of LAMP product with an FITC-labeled DNA probe that
confirmed the presence of specific, biotin-labeled WSSV amplicons. The resulting DNA duplexes could be visualized trapped
at the LFD strip test line within 5 min of sample exposure. Using the combined LAMP and LFD system, the total assay interval
was approximately 30 min, excluding 15 min of rapid DNA extraction time. Detection sensitivity was comparable to other commonly-used
methods for nested PCR detection of WSSV. In addition to reduced assay time when compared to electrophoresis,
combination of LAMP with LFD confirms amplicon identity by hybridization and eliminates the need to handle carcinogenic
ethidium bromide.

HISTORY AND STATUS OF PADDLEFISH AQUACULTURE IN CHINA
Hong Ji and Changzheng Wang
Department of Fisheries Science
Northwest A&F University
Yangling, Shaanxi 712100, PRC
jihong0405@hotmail.com
A total of 3,000 paddlefish (Polyodon spathula) larvae was first introduced into China from the United States in 1988. Importation
of fertilized paddlefish eggs has continued. In 2001, successful spawning of broodstock raised in China was achieved
in Hubei Province. Now, paddlefish is cultured in more than 10 provinces in China. The feeding habit of paddlefish is rather
similar to that of bighead carp, which is one of the major traditional fish species produced by Chinese aquaculture. Therefore,
Chinese fish farmers try to rear paddlefish as a substitute species of bighead in their production systems. Usually paddlefish
fingerlings (10 cm in TL) are cultured to market sizes in ponds or cages in reservoirs. Because of the limited supply of fingerlings
and the lack of equipment to catch paddlefish from large bodies of water, reservoir ranching as a production system has
not been developed. In ponds, paddlefish are reared in polyculture either as the primary species or as the secondary species.
When paddlefish is the primary species, the stocking rate of is 2400-4800 fishes/ha, and when grass carp are the primary species,
paddlefish are stocked at 900-3600 fishes/ha. In net cages, with the usual dimensions of 4m×5m×2.5m, suspended in
reservoirs, the stocking rates are 15-40 fishes per square meter if prepared diets are fed. The stocking rates are reduced to 1-5
fishes per square meters if the fish are not fed prepared diets and lighting is provided to attract zooplanktons as the natural diet
for paddlefish. There is still no specific diet designed for paddlefish. Currently, extruded feeds for snakehead, with a protein
level of 39% to 44%, is used for paddlefish culture. So far, paddlefish in China are cultured mainly for its meat rather than its
roes. The acceptable market size of paddlefish is 0.6-0.75 kg, which can be reached within six months if prepared diets are
provided; whereas, it takes about one year for paddlefish to reach the market size if no prepared diets are provided. Paddlefish
are usually marketed live since the processing industry has not developed. A few paddlefish are also raised and marketed as an
aquarium fish. There are only four paddlefish hatcheries in China, located mostly in Hubei Province. Together they can supply
only about 600,000 paddlefish fingerlings per year. Therefore, the short supply and high price of fingerlings are still the major
factors limiting the expansion of paddlefish production in China.
1 3

1 4
USE OF SOY BASED PRODUCTS IN PRACTICAL DIETS FOR WHITE SEABASS,
Atractoscion nobilis AND CALIFORNIA YELLOWTAIL, Seriola lalandi
Dave Jirsa*, Allen Davis, Mark Drawbridge and Kevin Stuart
Hubbs-SeaWorld Research Institute
San Diego, CA 92109 USA
djirsa@hswri.org
White seabass (WSB), Atractoscion nobilis and California yellowtail (YT), Seriola lalandi are two of the top five candidate
species for ocean farming in southern California. Hubbs-SeaWorld Research Institute (HSWRI) has maintained spawning
populations of WSB since 1984 and YT since 2003. WSB aquaculture has been directed primarily toward stock enhancement,
while YT is considered more for its commercial potential. Both species are carnivorous and are currently reared on standard
commercial diets whose protein ingredients have not yet been “optimized” relative to long-term sustainability issues associated
with fish meal and oil.
This project is consistent with several of the goals of the Plant Products in Aquafeed (PPA) Working Group Strategic Research
Plan, but most specifically to “establish standardized research approaches and protocols for systematic evaluation of plant
feedstuffs across carnivorous fish species”. Furthermore, recent California law (SB 201) states “To reduce adverse effects on
global ocean ecosystems, the use of fish meal and fish oil [in aquaculture] shall be minimized. Where feasible, alternatives to
fish meal and fish oil, or fish meal and fish oil made from seafood harvesting byproducts, shall be utilized . . .”
The specific goals of this project are to (1) develop and validate an open formulation for the WSB and YT based on modifications
of the existing commercial diet being fed to WSB and YT and (2) evaluate the response of WSB and YT to a diet with
increasing levels of soy protein concentrate as a replacement for fish meal, and (3) evaluate the response of WSB and YT to a
diet with increasing levels of soy oil as a fish oil replacement while keeping other nutritional factors constant.
The first trial is designed to validate our protocols and make a preliminary assessment of the species’ reaction to soy products
in the diets. For the first trial, WSB were stocked into 60L tanks supplied with 18°C recirculated seawater at a size of 3.5g and
fed 4-6% body weight daily. YT were stocked into 1000L tanks supplied with 21°C seawater at a size of 5.5g and fed 6-8%
body weight daily. Fish were weighed bi-weekly. After four weeks WSB weighed 7.3 – 8.2g and YT 18.7 -26.1g. Preliminary
results indicate that the open formulation feed performs similarly to current commercial diet and there is considerable potential
for fish meal to be reduced using either soybean meal and/or soy protein isolate.

AN EVALUATION OF CHANNEL CATFISH EGGS DERIVED FROM FISH HELD UNDER
DIFFERENT ENVIRONMENTAL CONDITIONS
Mini T. Jose and Alf Haukenes
Aquaculture / Fisheries Center of Excellence
University of Arkansas at Pine Bluff
Pine Bluff, Arkansas 71601 USA
mjose@uaex.edu
Spawning channel catfish and egg incubation practices have changed little over the past century but a current industry desire to
increase production of hybrid catfish fingerlings requires more controls over the spawning season. Timing of the channel catfish
spawning season can be varied by manipulating water temperature but may lead to variation in egg quality. The goal of this
experiment was to evaluate animals receiving a similar cumulative degree days achieved during a compressed versus a more
natural time frame. Three treatments varying the rate of increase of water temperature were applied to channel catfish in the
weeks before spawning: 1) a rapid (3 d) increase from 13 °C to ~24°C , 2) a gradual increase to 24-25°C over 5 weeks, 3) and
a natural temperature increase in ambient surface water temperature. After accumulating at least 168 cumulative degree days
we injected fish with carp pituitary extract to induce spawning (Figure 1). Ovulation rate and mean individual egg volume from
each group was determined. Sub-samples (~100 mL) of fertilized eggs were incubated in small hatching jars and fry survival
determined. Fertilized eggs from pond spawned fish were collected and compared to eggs produced from the fish held indoors.
Ovulation rates of the rapid, gradual, and natural temperature groups were 0, 21, and 84%, respectively (Table 1). Similar fe-
cundity (eggs/kg of female) was observed in ovulating fish from the gradually manipulated water temperature group and those
held under a natural temperature profile but average fry survival was 10 times higher in fish held under the normal temperature
profile. Total volume of individual eggs was significantly larger and higher fry survival was observed among pond spawned
Date
fish than fish held indoors. Degree days are useful in predicting reproductive maturation but continued exploration into proper
Figure 1. Degree days accumulated at spawning for three groups of
maintenance of broodstock held under artificial temperature regimes is warranted to decrease variation in egg quality.
Cumulative Degree Days
Rapid Gradual Natural
Date
Figure 1. Degree days accumulated at spawning for three groups of
channel catfish. Degree days were calculated as the average daily
temperature minus 18°C and summed. Arrows indicate spawning dates.
Table 1. Ovulation rates and mean (± SD) egg volume, yolk volume,
fecundity (eggs/kg) and percent fry survival of four groups of spawned
channel catfish held under different environmental conditions. *
Indicates significant difference form pond spawned and normal group.
See text for further description of treatments.
Rapid
(N=7)
Treatment Group
Gradual
(N=14)
Normal
(N=19)
Ovulation (%) 0 21 84
Pond
(N=5)
Fecundity
(eggs/kg)
NA 4747±2373 4726±1693
Yolk volume
(mm 3 )
NA 15.0±4.6 18.7±4.5 23.1±5.5
Egg volume
(mm 3 )
NA 24.3±5.6 25.7±0.0* 37.94±0.0
Fry Survival (%) NA 2.4±1.4 23.7±22.0* 41.1±9.2
Cumulative Degree Days
Rapid Gradual Natural
channel catfish. Degree days were calculated as the average daily
temperature minus 18°C and summed. Arrows indicate spawning dates.
Table 1. Ovulation rates and mean (± SD) egg volume, yolk volume,
fecundity (eggs/kg) and percent fry survival of four groups of spawned
channel catfish held under different environmental conditions. *
Indicates significant difference form pond spawned and normal group.
See text for further description of treatments.
Rapid
(N=7)
Treatment Group
Gradual
(N=14)
Normal
(N=19)
Ovulation (%) 0 21 84
Pond
(N=5)
Fecundity
(eggs/kg)
NA 4747±2373 4726±1693
Yolk volume
(mm 3 )
NA 15.0±4.6 18.7±4.5 23.1±5.5
Egg volume
(mm 3 )
NA 24.3±5.6 25.7±0.0* 37.94±0.0
Fry Survival (%) NA 2.4±1.4 23.7±22.0* 41.1±9.2
1

1
CLASSIFICATION AND QUANTIFICATION OF PHOSPHOLIPIDS AND DIETARY EFFECT
ON THEIR COMPOSITION IN SHRIMP (Litopenaeus vannamei)
Zhi Yong Ju, Ian Forster, Addison Lawrence and Warren Dominy
Aquatic Feed and Nutritional Department
Oceanic Institute
41-202 Kalanianaole Hwy,
Waimanalo, HI 96795 USA
zyju@oceanicinstitute.org
Phospholipids (PL) are essential dietary nutrients for shrimp. Soy lecithin is most often used as a dietary PL supplement in
shrimp feed formulations. Soy lecithin contains several classes of PL, including phosphatidylcholine (PC), phosphatidylethanolamine
(PE), phosphatidylinositol (PI), phosphatidic acid (PA), and phosphatidylserine (PS), as well as their lysophosphatidyl
derivatives. Limited information is available about the PL composition of Litopenaeus vannamei or the effects of diet on
PL classes present in shrimp whole body. This study examined the levels of different PL classes in L. vannamei whole body and
tail muscle, and evaluated how dietary PL composition affects the composition of PL and other lipids in shrimp body and tail.
Samples of ingredients, diets and shrimp were collected from an eight-week indoor trial with whole floc culture- or floc fraction-added
diets. The levels of PL, including PC, PE, PI, PA, and PS, were determined in the diets and shrimp samples by
HPLC. It was found that total PL comprised the majority of total lipid: >69.5% in shrimp tail muscle; >60.1% in whole shrimp
body. PC, PE, and PI were the most abundant PL classes, comprising over 40%, 35%, and 15%, respectively, of the PL in
shrimp tails. Cardiolipin (CL) and ceramides were also detected in shrimp body. An unidentified phospholipid was found in
marine-animal meals and in the shrimp. This unidentified phospholipid may be a unique in marine animals as biomarker phospholipid.
Diets with the highest (23.5 to 25.0 g/kg) and lowest (20.0 to 21.7 g/kg) total PL, as well as the highest (91.4 to 94.5
g/kg) and lowest (69.5 to 77.8 g/kg) total lipid, had no significant effect on major individual PL or total PL contents in shrimp
body. However, dietary crude protein content (40% vs 35%) affected PL composition of shrimp body (P

Macrobrachium rosenbergii NODAVIRUS (WHITE TAILED DISEASE) IN AUSTRALIA
Karen Juntunen*, Leigh Owens, Kathy La Fauce, Orachun Hayakijkosol, Kai Endo and Chaoshu Zeng
James Cook University
School of Veterinary and Biomedical Sciences
1 Solander Drive
Douglas, QLD 4811Australia
karen.juntunen@jcu.edu.au
The index case of white muscle disease presented in adult broodstock Macrobrachium rosenbergii from the Flinders River in
western Queensland in mid 2007. Histopathology revealed that the adults had extensive myonecrosis with massive
infiltration of myonuclei and some haemocytes. Subsequently, one family of juveniles from the same broodstock but not three
other families from different broodstock displayed white muscle lesions. Low grade chronic mortalities commenced.
Reverse transcriptase, polymerase chain reactions (RT-PCR) were attempted for both M. rosenbergii nodavirus (MrNV) with
two sets of primers and for the parasitic satellite virus, extrasmall virus (XSV). After modification and optimization, all PCRs
were positive for amplicons of the appropriate size. BLAST analyses of the consensus sequences gave the following results:
91 % match with Macrobrachium rosenbergii nodavirus segment RNA-2, complete sequence (Sekhar’s primers); 96% match
with Macrobrachium rosenbergii nodavirus RNA-directed RNA polymerase gene, partial codons (Yoganandham’s primers);
99% match with Macrobrachium rosenbergii XSV isolate M23 capsid protein gene, complete codons (Yoganandham’s XSV
primers).
It was shown that the clinical signs, histopathological lesions and RT-PCR amplicons could be reproduced in Macrobrachium
inoculated with cell-free extracts fulfilling Koch’s postulates. It is concluded that this is an endemic strain of MrNV as the
sequences are too dissimilar to be the same strain of MrNV circulating around Asia/Americas. It should be noted that this case
only poorly meets the Office International des Epizooties (OIE) case definition for white tail disease (WTD). We believe this
current case is clearly WTD, and that the OIE case definition needs broadening based on the information provided herein.
1

1
GENETIC CHARACTERISTICS OF THREE PADDLEFISH BROODSTOCKS BASED ON
MICROSATELLITE DNA ANALYSIS
Dariusz Kaczmarczyk* and Miroslaw Luczynski
Department of Environmental Biotechnology
University of Warmia and Mazury in Olsztyn
Słoneczna 45G
10-718 Olsztyn, Poland
d.kaczmarczyk@uwm.edu.pl
The American paddlefish has been introduced into Polish aquaculture in 1995 as a species suitable for breeding in polyculture
with common carp. The paddlefish may become a popular addition to Polish fish farms, as similar attempts in neighboring
countries have been successful.
In Poland, the paddlefish is reared in two farms: Pogórze and Wąsosze. Those stocks are foreseen as source of stocking material
for future paddlefish aquaculture. Stocks of Pogórze and Wąsosze consist of small number of fish, witch originated from
the same group of parental individuals and are vulnerable to the decline heterozygosity. In this study we have examined polymorphism
at 8 microsatellite loci in Pogórze and Wąsosze and in one Ukrainian stock (Gorny Tykich) as a reference. We have
evaluated genetic variation in Polish stocks and possibility of using the Gorny Tykich stock in order to enrich genetic pool of
the Polish stocks.
DNA was extracted from fragments of fin tissues taken from all fish belonging to Polish stocks (Pogórze 24 individuals,
Wąsosze 29 individuals) and from 47 individuals belonging to Gorny Tykich stock. All fish of the Pogórze stock were tagged.
The microsatellites and surrounding sequences were amplified using PCR technique. The length of the amplified fragments
was measured using automatic DNA sequencer Beckman Coulter CEQ 8000. Measures of observed (H o ) and expected heterozygosity
(H e ), allelic richness, bottleneck effect (Garza-Williamson index) and test of departures from H-W equilibrium were
performed. The genetic divergence between populations was estimated using two different parameters: F ST and δμ 2 . Individual
genetic characteristics were developed for all individuals from Pogórze stock and have been used in estimation of theoretical
proportion of heterozygotic genotypes in the progeny of each possible male x female combination. This procedure was applied
to a single locus and then was repeated for all loci. The results were averaged and obtained value was equal to the proportion
of heterozygotic individuals in the progeny of each spawning pair.
In the investigated stocks H o ranged form 0.56 to 0.60 and H e from 0.55 to 0.61; the values were similar to those specific for
native population inhabiting Kaskasia river (Ho 0.66, He 0.63). Most microsatellite loci were in the H-W equilibrium except
most polymorphic loci such Psp 20, Psp 26 and Psp 28. The mean M – value (from 0.40 in Wąsosze; to 0.48 in Gorny Tykich)
and number of alleles (Polish and Ukrainian stocks - from 29 to 33, Kaskasia river 55) suggested that genetic variation within
the investigated stocks was reduced because of the bottleneck effect. Ukrainian stock, witch was genetically distant to Polish
stocks (F ST value from 0.54 to 0.96) could be used to enrich of the genetic pools of Polish stocks. The distance between Polish
stocks was very small (F ST 0.007), what resulted from their common origin. Heterozygosity estimated for the progeny of
particular pairs of spawners from Pogórze stock varied from 0.25 to 0.68. The progeny’s expected heterozygosity was usually
below the value specific for their parent’s generation (0.55); only few parental combinations resulted in heterozygosity higher
than 0.55. The method presented in this study could be used to maintaining the genetic variation of Polish paddlefish stock at
a present level.

EFFECT OF TOTAL REPLACEMENT OF FISHMEAL WITH FERMENTED SOYBEAN AND
SQUID BY-PRODUCT BLEND IN PRACTICAL DIETS FOR CLIMBING PERCH Anabas
testudineus
Md. Abdul Kader*, Mahbuba Bulbul, Md. Sakhawat Hossain, Mohammad Mosarof Hossain, Gias Uddin Ahmed,
Md. Arshad Hossain, Saichiro Yokoyama, Manabu Ishikawa and Shunsuke Koshio
Laboratory of Aquatic Animal Nutrition
Faculty of Fisheries, Kagoshima University
Shimoarta 4-50-20
Kagoshima 890 0056, Japan
abdulkader_fc@yahoo.com
The continuous increasing demand for fishmeal (FM) in different agro-industrial sectors, environmental fluctuations and decreasing
wild stock of FM produced fish, made FM as limited and expansive resources which ultimately limit its further use in
aquaculture. In Bangladesh, now-a-days, quality fish meal is not available mainly because of high price which is beyond the
marginal fish farmer. Therefore, cost-effective, nutritionally balanced, environmentally sound and available alternative protein
feedstuff is necessary for sustainable aquaculture. A blend of animal and plant proteins might be a good approach for this purpose.
Due to the outbreaks of different diseases related to proteins from animal by-products, most of the animal proteins are no
longer available for use as animal feed. Sea food processing by-products still have importance to compensate this deficiency.
Among plant proteins, soybean meal is the most important for comparatively balanced nutritional composition, world wide
availability and low cost. Since efficient utilization of the alternative protein is necessary, fermentation process will be an interesting
approach to improve the quality of products.
The effects of total replacement of dietary FM with fermented soybean meal and squid by-product blend (7:3) (FSSq) on the
growth performances, feed utilization and profit indices of climbing perch (Anabas testudineus, Bloch, 1972) were investigated.
Five isonitrogenous and isolipidic diets were prepared by replacing 0, 25, 50, 75 and 100 % brown fishmeal protein with
FSSq. A commercial diet was also used as reference diet. The diets were tested using juvenile fish of 1.00 g initial weight, held
in triplicate groups (stocking density 40000 ha -1 ) of 30 m 2 earthen pond for 10 weeks. The test diets were hand delivered twice
a day at the rate of 10% of their body weight per day in the beginning and gradually decreased to 5% till end. At the end of
the feeding trial, growth performances, feed utilizations, whole body composition and economic benefits were analyzed. The
results of this research and impacts of the findings will be discussed.
1

1 0
FEEDING GOLDEN SHINER Notemigonus crysoleucas LARVAE FORMULATED DIETS
Thomas Kent*, Joseph Morris and Richard Clayton
Department of NREM
Iowa State University
Ames, Iowa 50011-3221 USA
thomask@iastate.edu
Golden shiners (Notemigonus crysoleucas) are popular baitfish for anglers as well as forage fish for other cultured fish in the
North Central Region. Since the original October 2006 USDA-APHIS VHS ruling that limited the interstate transport of specific
fish, there has been an increased demand for regionally reared baitfish. However, given this region’s short culture season
there is a need to investigate methods that produce market size, ca. 8 cm long, golden shiner fry in one culture season. The
purpose of this study was to use out-of-season spawning to obtain golden shiner fry, culture them intensively for 30 days, and
then stock them into earthen culture ponds for advanced growth.
Golden Shiner fry were obtained from broodstock that had spawned indoors on spawning mats. Eggs were removed from mats
by submerging the mats in a sodium sulfite solution for 2 minutes and then placing the eggs into 6-liter McDonald-type hatching
jars. The number of newly hatched fry were determined by volumetric displacement and subsequently stocked evenly into
100-L fiberglass tanks at random. Each pair wise comparison used three tanks per diet with the exception of one comparison
in 2007 when six tanks were used per diet. All tanks had in-tank lighting and water flow was 3 Lpm. Fry were fed at a rate of
5 grams/1,000 fry with auger feeders every 20 minutes for 22 hours per day with the exception of one diet that was hand fed
twice per day in 2008. In the two culture seasons, nine different diets were compared for their effect on fish survival; the best
performing diet in 2007 was used in 2008 comparisons. Each feeding trial was conducted for 14 days post hatch (dph), 10 dph
has been identified as the critical period for golden shiner fry survival. At the end of each 14-day test all surviving fry were
counted and survival determined.
At the completion of each 14-day trial, survival rates ranged from 0 to 16% in 2007 and 0 to 42% in 2008. The Zeigler® AP100
diet had the highest survival in one comparison with an average survival of 27.7% in 2008. All other diets had an average survival
that ranged from 0 to 8.3%, including four additional Zeigler® AP100 trials in both years. In 2007, both Zeigler® AP100
trials were found to have significantly greater survival than two diets in direct comparison (P

ON CHOICE AND USE OF STATISTICAL TOOLS IN EXPERIMENTAL FISH NUTRITION
STUDIES
Juhani Kettunen and Kari Ruohonen
Finnish Game and Fisheries Research Institute
Viikinkaari 4
FIN-00791 Helsinki
Finland
juhani.kettunen@rktl.fi
The main aim of the paper is to discuss and demonstrate the sequence of decisions that are necessary when using statistical
models in fish nutrition studies. We emphasize the key role of statistical experimental design that is, unfortunately, too often
flawed or completely missing in present studies. Furthermore, we demonstrate the choice of model that is based on the scale
and distribution of response and predictor variables. We recommend an iterative experimental strategy. Thus, instead of a single
one-shot experiment we prefer a set of consecutive, complementary experiments that are designed on the basis of existing
knowledge and outcome of the preceding trials. The philosophy behind is to start from screening the relevant variables with
simple and cost-effective designs and proceed towards more detailed understanding of the system with more targeted and often
more complicated designs. We speak of model-based design and analysis of fish nutrition problems. By doing this we design
our experiments to minimize the posterior uncertainty of the model parameters and forecasts. For this the choice of a model on
the basis of the scale and distribution of response and predictor variables is crucial. In the paper, we compare the model-based
approach with the traditional design-based approach that emphasizes statistical hypothesis testing in finding significance of effects.
We demonstrate many strengths of model-based approach in comparison with design-based one. Most important strength
of the model-based strategy is the information that is given to decision makers, i.e. users of the results. While the design based
approach gives him/her a on-off solution, the model-based leaves the final decision to the decision maker. This is possible,
because also estimates of the risk and uncertainty levels are produced.
1 1

1 2
THE EFFECT OF A PREBIOTIC SHORT CHAIN FRUCTOOLIGOSACCHARIDE (scFOS)
ON THE GROWTH, SURVIVAL, AND IMMUNE RESPONSE OF ADULT PACIFIC WHITE
SHRIMP Litopenaeus vannamei UNDER CONDITIONS OF PRIMARY PRODUCTIVITY
Brandon C. Klim*, Addison L. Lawrence, Susmita Patnaik, Joe M. Fox, Bart R. Dunsford and David A. McKee
Texas AgriLife Research Laboratory
Texas A&M System
Port Aransas, TX 78373 USA
bklim@islander.tamucc.edu
Prebiotics have been shown to enhance the health, growth and survival of many terrestrial animals and have become a common
feed supplement in livestock, pet and human foods providing a number of nutritional benefits. These benefits are attributed
to the selective nature of prebiotics in promoting the growth of beneficial bacteria. These bacteria out-compete harmful bacterial
and carry out metabolic activities that are advantageous to the host. Recent studies have shown that supplementation
of a short-chain fructooligosaccharide (scFOS) prebiotic in shrimp feed significantly alters the microbial flora of the gut and
enhances immune response in juvenile Pacific white shrimp under clean water conditions. The present study explores the potential
benefits of this prebiotic by applying it to grow-out of adult Pacific white shrimp in the presence of primary productivity.
These conditions are thought to more closely resemble those of a typical commercial shrimp farm and provide commercially
applicable information on the use of this prebiotic.
A 10-week growth trial with adult Pacific white shrimp is being conducted outdoors in 2,180L (2.35m diameter x 0.50m depth)
tanks with a water exchange rate of 33%/d and stocking density of 75 shrimp/m 2 (150/m 3 ). A growth trial using diets with
0.08% scFOS and without scFOS at feeding rates of 1.0, 1.3 and 1.6 g/shrimp/wk is currently being undertaken with five replicates
for each dietary treatment. Shrimp were stocked at a mean weight of 5.23g and are being reared under ambient salinity
and temperature conditions.
Upon termination of the study, mean survival and growth of shrimp as well as feed conversion ratio will be determined. In
order to quantify immune response, haemolymph will be extracted from subsamples of shrimp from each tank. These analyses
will include total haemocyte count, haemocyte respiratory burst and phenoloxidase activity.
Survival, growth and FCR will be analyzed statistically for significant differences between treatments with and without scFOS
as well as among different feed rates. Immune response data will be analyzed and compared among treatments to provide
supporting evidence corresponding to possible survival and health benefits. The results of this study will help to demonstrate
whether significant survival, growth and immune benefits can be achieved using scFOS as a prebiotic supplement to commercial
shrimp feed. Research findings will be discussed in the paper.

TISSUE DISTRIBUTION OF LEPTIN-LIKE MRNA AND CHANGES IN ITS EXPRESSION
AFTER PROLONGED FASTING AND EXPOSURE TO Edwardsiella ictaluri IN CHANNEL
CATFISH
Yasuhiro Kobayashi, Sylvie Quiniou and Brian C. Peterson
Division of Biological and Physical Sciences
Delta State University
Cleveland MS USA
Leptin is thought to be a key pleiotropic cytokine involved in regulation of food intake and energy homeostasis in fish and
mammals. Studies show that nucleotide and amino acid sequence of leptin are poorly conserved among different fish species
and share very little similarity with the mammalian counterpart. An mRNA transcript that encodes leptin-like peptide has been
identified in channel catfish. However, the function of leptin in channel catfish is unknown.
The objectives of this research were to characterize tissue expression of leptin-like mRNA and changes in its expression after
prolonged fasting and exposure to Edwardsiella ictaluri in channel catfish. Catfish leptin-like gene was sequenced by genome
walking. The coding sequence for catfish leptin-like gene was determined by 5’- and 3’-RACE. Our results showed that the
open reading frame of the catfish leptin-like mRNA was 519 bp and encoded a peptide that was 172 amino acids in length.
Analyses of the secondary structure revealed a series of helixes and coils that were conserved among peptides that belonged
to the leptin superfamily. Putative amino acid sequence of the channel catfish leptin-like peptide shared very low sequence
similarities with leptin of other fish species or the mammalian leptin (24-49 %). Leptin-like mRNA expression was detected
in various tissues including brain, stomach, spleen, heart, liver, and trunk kidney but was especially high in liver and trunk kidney.
Expression of leptin-like mRNA in liver and brain was unaffected (P > 0.1) by prolonged fasting or 15 days of refeeding
after prolonged fasting. Hepatic expression of leptin-like mRNA was elevated (P < 0.05) 24 hours after fish were exposed to
E. ictaluri. In comparison, expression of leptin-like mRNA in trunk kidney and spleen was elevated (P < 0.05) 48 hours after
E. ictaluri exposure. Furthermore, spleen expression of leptin-like mRNA was elevated (P < 0.05) 72 hours post exposure to
E. ictaluri.
Based on our results, catfish leptin-like gene does not seem to assume mammalian leptin functions. Further investigations are
required to clearly define the identity and functions of this transcript.
1 3

1 4
THE U.S. FDA’S THIRD PARTY CERTIFICATION PILOT OF AQUACULTURED SHRIMP
Brett Koonse
U.S. Food and Drug Administration
Center for Food Safety and Applies Nutrition
College Park, MD 20740 USA
brett.koonse@fda.hhs.gov
On July 10 th , 2008 The Food and Drug Administration (FDA) announced an opportunity for third party certification programs
or certifying bodies (including those that are private, non-government entities that are independent of the businesses they audit,
other Federal government, State government, and foreign government agencies and officials) who certify aquaculture shrimp
farming and/or processing facilities for being in compliance with FDA or equivalent food safety rules or regulations, to apply
to participate in a voluntary third party certification pilot for aquaculture shrimp which was conducted by FDA’s Center for
Food Safety and Applied Nutrition (CFSAN) and Office of Regulatory Affairs (ORA).
This talk will update Aquaculture America on the status of this pilot program and where FDA is headed in using third party
certification bodies to help assure the safety of the U.S. food supply.

PALM OIL EFFICACY IN OXIDATION STABILITY WITH OXIDIZED FISH OIL ON
GROWTH, FATTY ACID COMPOSITION AND OXIDATIVE STRESS CONDITIONS IN RED
SEA BREAM, Pagrus major, JUVENILES
Shunsuke Koshio*, Connie F. Komilus, Manabu Ishikawa and Saichiro Yokoyama
Aquaculture Group
Faculty of Fisheries
Kagoshima University
Japan
This study was aimed to investigate the effects of replacing fish oil with palm oil (P) blended with oxidized fish oil (F) as the
lipid source on growth performance, lipid profiles and oxidative stress condition of red sea bream. A 50-day feeding trial by
using juveniles with initial body weight of 5.2 g was conducted in triplicates using 8 isocaloric diets containing two degrees of
oxidized fish oil (peroxide values of 2.2 and 86.8 meqkg -1 ) blended with palm oil at different levels. Formulated experimental
diets used consist of 100% fish oil (1F:0P), 60% fish oil + 40% palm oil (3F:2P), 50%fish oil + 50% palm oil (1F:1P), 40% fish
oil + 60% palm oil (2F:3P) with and without oxidized fish oil. POV values of the test diets containing oxidized fish oil were
21.3, 21.8, 20.4, and 22.1 for 1F:0P, 3F:2P, 1F:1P, and 2F:3P, respectively.
Both oxidized oil and palm oil level were significant factors on body weight gain, and the values of fish fed the diets with oxidized
1F:1P and oxidized 2F:3P were significantly lower (P

1
CORRELATION OF ULTRASOUND IMAGING WITH HISTOLOGICAL ASSESSMENT AND
CONDITION FACTORS IN THE DEVELOPMENTAL STAGING OF CHANNEL CATFISH
OVARIES
Douglas E. Kuenz*, Noel Novelo, Christopher Green and Terrence R. Tiersch
Aquaculture Research Station
Louisiana State University Agricultural Center
2410 Ben Hur Rd.
Baton Rouge, LA 70820 USA
dkuenz1@lsu.edu
Before, during, and after spawning season, multiple stages of gonad maturation can be found in populations of channel catfish.
The purpose of this study was to compare ultrasound imaging with histological assessment and several condition factors to
better define stages of gonadal maturation of female channel catfish. Fish were studied from April through July, 2008 in Baton
Rouge, Louisiana. In total 51 females were sampled before, during, and after the spawning season to ensure a wide range of
developmental stages for analysis.
An ultrasound image of the ovary was captured and classified from “Stage 1” (undeveloped) to “Stage 6” (atretic), with “Stage
5” being fully mature and ready to spawn. Females were selected randomly from each of the ultrasound groupings to ensure
representation of all stages in the histological and condition factor assessments. Attributes recorded were length, weight, girth,
gonad weight, liver weight and visceral fat weight, from which were calculated several indices including hepatosomatic index,
gonadosomatic index and condition factor (Table 1).
Photographs were used to record condition of fish, livers, and gonads. Sections were taken from the middle of the ovary to
represent the area viewed by ultrasound. These were fixed in 10% buffered formalin and stained with hematoxylin and eosin.
Oocyte maturation was classified from “Stage 1” (pre-vitellogenic) to “Stage 6” (atretic) with “Stage 5” representing readiness
to spawn.

A SPREADSHEET MODEL FOR FINANCIAL PLANNING OF TILAPIA AND AFRICAN
CATFISH BUSINESSES
Ganesh Kumar*, Nathanael Hishamunda and Carole Engle
University of Arkansas Pine Bluff
1200 North University Drive
Mail Slot-4912
Pine Bluff, AR 71601 USA
gkumar@uaex.edu
Detailed financial planning for aquaculture businesses can make the difference between profits and losses especially in years
with unfavorable prices and costs. However, many small and medium scale farmers throughout the world lack the knowledge
to develop comprehensive financial statements and may not have access to accountants for assistance. User-friendly decisionmaking
tools that allow small and medium-scale farmers to assess profitability levels in aquaculture ventures may contribute
to successful business development. FAO, in collaboration with the University of Arkansas at Pine Bluff, has developed a userfriendly
decision making spreadsheet model for financial planning for Tilapia and African catfish farms. The model contains
modules for four basic types of culture systems: pond farming, cage culture, tank recirculatory systems and hatchery rearing
systems. The user-friendly model allows the farmer to enter farm details, production parameters, input and investment costs.
Default values are included in the event that the user does not know some values. The model will then develop an enterprise
budget, breakeven analysis, sensitivity analysis, balance sheet and, income statement. It also calculates solvency and liquidity
indicators, and an amortization schedule. An electronic version of the model as well as a guidance manual is available upon
request from FAO.
1

1
“SELF-CLEANING MICROCOSM TANKS” AN APT REARING SYSTEM FOR TESTING
FEEDS WITH SHRIMP IN A SIMULATED POND ENVIRONMENT
Kumaraguru Vasagam, K.P., Victor Suresh, A. and George W. Chamberlain
Integrated Aquaculture International
3303 West Twelfth Street
Hastings, NE 68902-0609 USA
drvasagamguru@gmail.com
Glass or fiberglass tanks using flow-through or re-circulated water are widely used in shrimp feeding experiments. Such
systems, however, do not adequately mimic the commercial shrimp culture systems which are abundant of natural foods. Few
studies used microcosm tanks for testing feeds, and the results are more relevant to commercial conditions. But they were
missing a system to closely monitor the feed consumption and discard the accumulated faeces and other organic matter. At
the Shrimp Nutrition Research Center (SNRC), Integrated Aquaculture International, USA, and the Department of Fisheries,
Brunei Darussalam, have jointly conducted feeding trials with Litopenaeus stylirostris in microcosm tanks since early 2007.
The microcosm tanks were cylindrical, free standing
fiber glass tanks of 1827 L water volume. Each
microcosm tank has a self-cleaning mechanism
driven by airlift and gravity. The swirling water
movement generated by airlifts in the inner periphery
of tanks sweeps all the solid particles (uneaten
feed, feces, plankton die-offs etc.) on the bottom
into the central cup and airlifted to the particle trap
on the top of the tank. While a 12 hour baby belt
feeder feeds the shrimp continuously, the excess
uneaten feed along with other solids at the bottom
are swept to the self cleaning trap at the top of
microcosm tank.
In three eight-week feeding trials, juvenile shrimp
of 0.9-4.3 g were stocked at a density of 28 shrimp/
m 2 . At the end of each trial, survival rates exceeded
80%. Growth rates ranged from 1.19 to 2.46 g/week.
Water quality remained stable and within suitable
ranges for L. stylirostris growth in all trials. The
tanks had algae and bacterial floc developing within
a few days of starting the trials. Fourteen commercial
shrimp feeds, each containing more than 40%
crude protein, were tested in the trials. In spite of
the presence of natural food organisms, significant
feed-related differences (Figure 1) among treatments
were found in each trial. In conclusion, microcosm
tanks support excellent growth and survival of
L. stylirostris and are appropriate for conducting
trials to evaluate feeds for pond growout.

SUSCEPTIBILITY OF PACIFIC SALMONIDS, YELLOW PERCH, AND KOI TO VIRAL
HEMORRHAGIC SEPTICEMIA VIRUS STRAINS FROM THE GREAT LAKES, PACIFIC
AND ATLANTIC COASTS OF CANADA, AND EUROPE
Gael Kurath*, Evi Emmenegger and Chang Hoon Moon
USGS Western Fisheries Research Center
6505 NE 65th St.
Seattle, WA 98115 USA
gkurath@usgs.gov
The emergence of viral hemorrhagic septicemia virus (VHSV) in the Great Lakes region has resulted in dramatic multi-species
epidemics since 2006. Although the virus is well known in Europe and on the Pacific coast of North America, this is the first
time VHSV has caused large-scale epidemics in freshwater fish. To date VHSV has been isolated from free-ranging stocks of
over 25 fish species in the Great Lakes, and it has also spread to inland lakes and river systems. To better understand the threat
of the Great Lakes VHSV to aquaculture species, we are comparing susceptibility of yellow perch, Chinook salmon, rainbow
trout, and koi to four different strains of VHSV. The virus strains were chosen to represent VHSV from the Great Lakes (type
IVb), brackish water sites in New Brunswick (type IVb), the Pacific coast of Canada (type IVa), and European trout farms (type
1). Fish were infected by intraperitoneal injection with both low and high challenge doses or each virus strain, and clinical
disease signs and mortality were monitored for 3-4 weeks. In Chinook salmon, the Great Lakes VHSV strain is more virulent
than the Pacific coast marine strain, but less virulent than European virus strain. Our results in all species show variation in
virulence of the different virus strains, indicating biological differences relevant to managing the virus if it appears in aquaculture.
1

1 0
THE INFLUENCE OF LYSINE DEFICIENCY ON YELLOW PERCH (Perca flavescens)
FEMALES REPRODUCTIVE PERFORMANCE AND THE QUALITY OF EGGS
Karolina Kwasek *, Konrad Dabrowski, Joanna Nynca, John M. Reddish and Macdonald Wick
The Ohio State University
Department of Animal Sciences
2029 Fyffe Court
Columbus, Ohio, 43210 USA
kwasek.1@osu.edu
We have previously shown that yellow perch fed wheat gluten protein-based diets supplemented with free lysine, (+) Lys diet,
achieved significantly larger mean weights than yellow perch fed a diet deficient in lysine (- Lys) (Kwasek. et al. 2008). We
have also shown that lysine deficiency has an impact on overall gametogenesis and frequency of body deformities of yellow
perch broodstock, and influences the maturation and sperm quality in this species. The objectives of the present study were:
(1) to determine whether Lys influences the ovulation of yellow perch, and (2) to evaluate whether Lys has an impact on free
amino acid pool of yellow perch eggs.
Yellow perch juveniles raised on formulated commercial diets at an initial size of 12.9 ± 4 g were individually marked with
pit-tags and randomly distributed into six 400 L tanks, 45 fish per tank. This experiment included two diets: (-) Lys-deficient
and (+) Lys-supplemented (2.23 % dry basis) wheat gluten-based diets in triplicates. Control females were fed a commercial
diet. Females from control, +Lys and –Lys groups that had ovulated were stripped and their eggs divided into 0.4-1.2 g (0.77 ±
0.11 g) portions and mixed with sperm (21.4 ± 4.3 μl). Samples of eggs were weighed, counted and stored.
Our results showed that the mean weight of females fed experimental diets and their relative egg weight ratio (Fig.1.) were
not significantly different. Females from +Lys group had significantly larger condition factor than those from –Lys group. The
largest amount of eggs was obtained from control fish in comparison to fish from the other treatment groups. The mean survival
was higher in eggs from control and +Lys females (15.2, 13.8 %, respectively) in comparison to –Lys females (5.13 %). The
free amino acid levels in eggs remain to be evaluated.

ALTERNATIVE USES FOR ALGAE PRODUCED FOR PHOTOSYNTHETIC CO 2
MITIGATION
Louis Landesman
128 Lakeview Park Road
Colonial Heights, VA 23834 USA
llandesman@vsu.edu
The burning of coal for electricity production entails the release large amounts of CO 2 , a gas that contributes to global warming.
In addition significant amounts of nitrogen and sulfur oxide gases are also released. Greenfuel and other startup companies
have pioneered in the application of algae production to coal combustion that captures most of this exhaust CO 2 along with the
nitrogen and sulfur oxides and converts these gases into algal biomass. This algal biomass is presently being dried and used as
fuel together with coal to produce electricity. This paper discusses the technical and commercial feasibility of using this algal
biomass for animal or fish feed as well for other uses, such as a ink pigment, paint, food coloring and for new pharmaceutical
compounds. Finding commercial uses of this biomass will improve the economic feasibility of carbon dioxide sequestration
by algae.
1 1

1 2
COMMERCIAL STANDARD PRACTICAL RESEARCH FEED FOR THE PACIFIC WHITE
SHRIMP, Litopenaeus vannamei
Addison L. Lawrence*, Susmita Patnaik, Warren Dominy, Ian Forster, Tom Zeigler and Tim Markey
Texas AgriLife Research Mariculture Laboratory at Port Aransas
Texas A & M System
1300 Port Street
Port Aransas, TX 78373 USA
Phone: 361-749-4625
Fax: 361-749-5756
smpall@yahoo.com
A commercial standard practical research feed is needed for conducting production studies in ponds and raceways, to evaluate
feed mill production methods effect on factors including feed physical and nutrient characteristics. A five week growth trial
was conducted to determine the growth and survival response to a proposed commercial standard practical research feed for the
Pacific White shrimp L. vannamei. A standard semi-purified research feed for the Pacific White shrimp L. vannamei has previously
been reported to support growth of L. vannamei from approximately 10 g to 30 g in size at a growth rate of 2.33 g/wk
and over 90% survival and has outperformed Rangen 35% and 40% commercial feeds by 20%. The study was conducted in a
recirculating indoor tank system (recirculation rate = 1636%/day and exchange rate = 25.8%/day) consisting of 102 rectangular
tanks (bottom area = 0.333 m 2 ) with L. vannamei at an initiation size of 0.5 g (SPF, Kona, Oceanic Institute, Hawaii). The
stocking density was 61 shrimp/m 2 . All treatments were fed at 2 g/shrimp/wk, 15 times a day by hand. All tanks were aerated
to maintain a minimum oxygen level of 5 ppm. The experiment was conducted at ambient water salinity and at a temperature of
29-31 °C. Both treatments had five replicates each. Total Ammonia Nitrogen (TAN), nitrite, nitrate and pH were monitored on a
weekly basis. At the study termination, shrimp were harvested and weighed for each tank separately. Water quality, and shrimp
harvest data including survival, growth, biomass and FCR were analyzed for statistical significance between the treatments.
Shrimp survival was 92.0 % to 88.0% for the standard commercial semi-purified diet and commercial standard practical diet,
respectively. Final weights were 8.57 and 7.99 g with estimated growth rates in the linear growth phase (above 3.5 g/shrimp) of
2.3 and 2.2 g/wk, respectively. These data showed that a growth and survival rate for the proposed commercial standard practical
research feed was not significantly different from growth and survival of the standard semi-purified research feed. Since
the standard semi-purified research feed has been showed to provide outstanding growth and survival to approximately 29-30 g
in size these data indicate that this commercial standard practical research feed would be very adequate to be used for raceway,
pond and tank productions and feed mill research. This commercial standard practical research feed is an open formulation.

DEVELOPMENT OF SPF STOCKS OF BAIT SHRIMP Litopenaeus setiferus AND
DEMONSTRATION OF LOW COST MATURATION AND LARVAL SYSTEMS
Alisha D. Lawson*, Jason Haveman, John W. Leffler, Beth L. Thomas, Jesus A. Venero, Al D. Stokes
and Craig L. Browdy
SCDNR - Waddell Mariculture Center
211 Sawmill Creek Rd.
Bluffton, SC 29910 USA
LawsonA@dnr.sc.gov
A NOAA National Marine Aquaculture Research Initiative has focused on producing specific pathogen free broodstock of
Litopenaeus setiferus and Farfantepenaeus duorarum. At the Waddell Mariculture Center (WMC) in Bluffton, South Carolina,
F1 nauplii were produced from artificially inseminated spawners collected off the coast of Charleston SC in 2006 and 2007.
Following disease testing and larval culture, broodstock were produced in WMC ponds and distributed to commercial postlarvae
producers. During 2007, several families were reared individually, tagged and grown out to broodstock size to establish a
founder stock.
Present research at the WMC has focused on demonstrating low cost small scale maturation and hatchery production systems.
F2 SPF L. setiferus broodstock were maintained in three 12 ft diameter indoor tanks. The shrimp were separated by sex with
two tanks containing 75 females and a third tank stocked with 75 males. The unisex system was operated according to (Browdy
et al. 1996). Tanks received UV treated fresh seawater (72% per day) and were linked by a recirculation system (145% exchange
per day) incorporating particulate, biological and UV filtration. Temperature in maturation systems was maintained
at 25-26 C. by heating or chilling incoming seawater to minimize potential for male reproductive tract melanization. Shrimp
were fed frozen adult Artemia, squid, marine polychaetes, and a maturation diet (Zeigler Bros, Garners, PA). Photoperiod was
manipulated using dawn/dusk simulators (light from 11:30 p.m. to 10:30 a.m.). Beginning 2 weeks after eyestalk ablation, each
morning females with mature ovaries were sourced from all female tanks and moved to all male tanks for mating as necessary.
The females were checked for mating 90 minutes after dusk and moved to covered spawning tanks. Within 24 hours, females
were placed back into their tank of origin and fertile spawns were harvested and moved into larval rearing tanks. Maturation
and spawning efforts produced 4 million nauplii over a period of 3.5 months and little or no signs of male reproductive tract
degeneration were observed in the male tank where females had been introduced for mating.
A larval rearing system was built using six 1000 L fiber glass tanks and two 10,000 L water treatment reservoirs. Larvae were
fed a mixed diet, based upon larval stages, of an algal species of Chaetoceros, frozen or live Artemia nauplii, microcapsulated
liquid larval diet and dry larval diets (Zeigler Bros Inc. At PL 7 to PL 12 , larvae were harvested and distributed for bait shrimp
production trials. These efforts have established founder stocks and demonstrated technologies necessary for commercialization
of US bait shrimp production.
1 3

1 4
NUTRITIONAL STUDIES ADVANCING ATLANTIC STURGEON BROODSTOCK
DEVELOPMENT AND RESTORATION SCIENCE
Andrew M. Lazur* and Erin Markin
Horn Point Laboratory
University of Maryland Center for Environmental Science
Cambridge, MD 21613 USA
alazur@hpl.umces.edu
One facet of a collaborative Chesapeake Bay Atlantic sturgeon restoration program involves studies related to sturgeon nutrition
and feeding management practices. Feed training of wild-caught fish is being evaluated to develop feeding protocols in
support of establishing a captive broodstock population. Other studies are evaluating fatty acid enrichment of live larval foods
and pre-release weaning assessment, to advance the technologies applicable to future replenishment research. A summary of
findings and implications for future research will be summarized.
Two feed training methods, force feeding and passive introduction of natural foods, were evaluated. After initial introduction
of natural foods, varied ratios of natural and commercial pellets in several forms were used to transition them to a diet consisting
entirely of commercial pellets. Fifty-eight of sixty-seven fish (85%) have been successfully trained to a commercial pellet
during year one of this study. Seven fish did not train to natural foods, while two fish transitioned to various passive diets but
did not survive until acceptance of commercial diet, most likely due to excessive weight loss. Both methods were successful in
training sturgeon, however, force feeding was observed to be stressful and unnecessary due to the success achieved with various
passive feeding regimes. After successful acceptance of commercial diets, fish were observed to be lean. In addition, the
inability to obtain viable sperm from successfully feed trained fish indicates possible holding stress and nutritional deficiencies
and warrants further studies.
Fatty acid enrichment of Artemia was conducted using three commercially available algal based enrichments (10% DHA, 10%
DHA and astaxanthin, and ARA) and fed to eight day old Atlantic sturgeon fry. Survival ranged from 72-99%. Mean weights
were 0.27, 0.26, 0.24, and 0.17 grams for the ARA, control, 10% DHA and astaxanthin, and 10% DHA respectively. Though
some difference in growth was observed between enrichments, growth in the control or non-enriched treatments was similar to
two of the enrichments supporting that enrichment is not necessary.
The ability of hatchery-reared fish to accept natural foods in released sites is an important question in developing replenishment
plans. To evaluate this, a brackish water closed pond study evaluated two juvenile year classes’ ability to adapt to natural foods.
Fish were PIT tagged, divided into two treatments: weaning from commercial diet to natural foods prior to release, and released
without weaning; and stocked into a 0.2 hectare pond supplied with natural foods. Significantly greater specific growth rate
was observed in the weaned fish of both year classes suggesting a benefit of weaning. Future studies will involve younger fish
in freshwater environments.

GROWTH PERFORMANCE OF ATLANTIC COD Gadus morhua LARVAE FED SQUID
HYDROLYSATE PREPARED AT DIFFERENT HYDROLYSIS TIMES
Chong M. Lee*, Qing Pan, Cetin Cetiner and David A. Bengtson
Dept. of Nutrition and Food Sciences
University of Rhode Island
Kingston, RI 02881 USA
chonglee@mail.uri.edu
Squid (Loligo pealei) is currently the most abundant and highest revenue-generating resource in Rhode Island. Squid processing
plants in the region generate about 4,500 metric tons of processing byproduct waste annually. The squid byproduct (Loligo
pealei), composed of heads, viscera, skin, fins, and small tubes, was converted into concentrated hydrolysate (22% solids,
17.3% protein and 3.0 % lipid, primarily phospholipids - 11.6% EPA/ 24.5% DHA on a lipid weight basis). It was hypothesized
that growth and survival of larvae could be affected by feeding larval diet with hydrolysate prepared by different degrees of hydrolysis,
and the degree of hydrolysis should be gradually lessened with the phase of larval growth as larvae develop a gut and
are able to digest more complex proteins. Squid hydrolysates (SH) of two different degrees of hydrolysis were thus prepared at
1 and 2 h at 55°C and incorporated into microparticulate diets as a sole protein source (73% of the whole diet; 56.4% protein,
24.7% lipid, 7.3% carbohydrate and 19.97 MJ/kg, on a dry weight basis). Microdiets were prepared using drum and plate drying
and micronized to 300 µm and 500 µm.
Atlantic cod (Gadus morhua) larvae of 30 d.a.h. previously fed rotifer and Artemia were brought into aquaria at 10°C (20 fish
each in triplicate) and acclimated for 2 wk with DHA-enriched Artemia. Thereafter, larvae were on 3 different microdiets for 6
wk including Artemia for initial 3 wk as a control. Based on the initial 3 wk feeding trial, the growth performance on microdiets
was comparable to that on the Artemia control. After 3 wk feeding period, Artemia was not able to support the normal growth
as larvae passed the weaning phase. The growth performance was assessed in terms of body weight (BW), body length (BL),
body condition factor (BCF) and specific growth rate (SGR). For 6 wk feeding, 63-68% of larvae survived through the weaning
period. Results of growth performance are shown in the table below. No significant differences (p>0.05) in all parameters
studied for growth performance among microdiets prepared with SH of different degrees of hydrolysate (1 h, 2 h hydrolysis
and 2h hydrolysis diet followed by 1h hydrolysis diet). This suggests that 1h hydrolysis was sufficient for SH to be digestible
to larvae and no added benefits are expected from extended hydrolysis. In addition, a two-stage feeding regimen with 2 h SH
diet (more hydrolyzed and lower m.w. peptides) followed by 1 h SH diet (less hydrolyzed and higher m.w. peptides) did not
demonstrate expected benefits as hypothesized.
Growth performance of cod larvae during a 6 wk-feeding on squid hydrolysate microdiets
Diets IBL cm IBW g IBCF FBW (g) FBL (cm) FBCF Survival
SH2 1.76±0.14 0.04±0.01 0.72±0.07 0.505±0.04 4.01±0.08 0.77±0.02 67.7±27 5.99±0.10
SH2-1 1.76±0.14 0.04±0.01 0.72±0.07 0.474±0.01 3.88±0.03 0.79±0.01 65.0±8.7 5.85±0.03
SH1 1.76±0.14 0.04±0.01 0.72±0.07 0.517±0.06 4.04±0.12 0.78±0.04 62.9±12 6.19±0.33
SH1: 1 h hydrolysate microdiet for 6 wk feeding; SH2: 2 h hydrolysate; SH2-1: 2h hydrolysate for 3wk followed by
1 h hydrolysate for 3 wk; IBW (FBW): initial (final) body weight; IBL(FBL): initial (final) body length; IBCF
(FBCF): initial (final) body condition factor; SGR: specific growth rate
(%)
SGR
1

1
EVALUATION OF THE DIETARY TOXIC LEVEL OF SELENIUM IN GROWING OLIVE
FLOUNDER Paralichthys olivaceus
Seunghyung Lee, Jun-Ho Lee, Jun-Young Bae, Young Chul Kim, Okorie E. Okorie, Gun Hyun Park
and Sungchul C. Bai*
Dept. of Aquaculture/Feeds and Foods Nutrition Research Center
Pukyong National University
Daeyeon-3-dong
Nam-gu, 608-737 Rep. of Korea
scbai@pknu.ac.kr
This feeding trial was conducted to evaluate the dietary toxic level of selenium in growing olive flounder, Paralichthys
olivaceus. Fish averaging 31.5 ± 0.46 (mean ± SD) are being fed one of the seven semi-purified diets containing 0, 2, 4, 6, 8,
10 and 20 mg Se/kg diet for 16 weeks. Growth performance, serological and hematological characteristics, histopathological
analyses and whole-body proximate composition will be discussed later.

IDENTIFICATION, CLONING AND EXPRESSION OF AGARASE FROM MARINE
BACTERIA STRAIN AG21
Youngduk Lee, Chulhong Oh, Ilson Whang, Se Jae Kim and Jehee Lee
Department of Marine Biotechnology
College of Ocean Science
Cheju National University
Jeju, 690-756, Republic of Korea
Agar is an important gelatinous substance present in the cell wall of some marine red algae (Rhodophyceaea). It is mainly
composed of agarose and agaropectin. Agarose is a hydrophilic hetero polysaccharide, which consists of a linear chain of alternately
arranged 3-O-linked β-D-galacto-pyronose and 4-O-linked 3,6-anhydro-α-L-galacto pyronose residues. Agarases are
the natural enzymes of certain agarolytic organisms found mostly in marine habitats that hydrolyze the agarose. Also, agarase is
the first enzyme in the agar catabolic pathway. Availability of agarases basically in the marine environment, which is consistent
with the fact that agar, being a product of marine algae is available to utilize some marine organisms as a convenient carbon
and energy source.
We screened agarase producing marine bacteria from Jeju sea. We isolated genomic DNA from the bacteria and analyzed 16s
rRNA sequence by PCR and sequencing. The sequence was 99% (1376/1384bp) similar to Saccharophagus degradans 2-40.
We amplified patial sequence from the strain by PCR. The detection of agarase full sequence, was carried out long and accurate
polymerase chain reaction (LA-PCR). The ORF was detected as 1908 bp (636 aa) and nucleotide sequence was showed 88%
similarity to Saccharophagus sp. agarase.
We tested optimum temperature, optimum pH and thermostability. Optimum temperature and pH were 55°C and 7.5 respectively.
The purified agarase was stable at 40°C for 30 hours. Agar degrading pattern by purified agarase was identified with
thin layer chromatography.
1

1
MONITORING TOOLS FOR CHARACTERIZING AND MANAGING MICROBIAL BIOFLOC
COMMUNITIES IN SUPERINTENSIVE ZERO-EXCHANGE SHRIMP CULTURE SYSTEMS
John W. Leffler*, Alisha Lawson, Andrew Ray, Gloria Seaborn, Andrew Shuler, Beth Thomas, Jesus Venero,
Luis Vinatea, and Craig L. Browdy
South Carolina Department of Natural Resources
Marine Resources Research Institute
217 Fort Johnson Road
Charleston, SC 29412 USA
Lefflerj@dnr.sc.gov
Superintensive, zero-exchange shrimp culture technologies demonstrate great potential for commercialization. Such systems
rely on the structure and function of a rich microbial “biofloc” community to process nitrogenous wastes, stabilize water
chemistry, and provide supplemental nutrition to the shrimp. However biofloc communities differ and may achieve these goals
to varying degrees. We have evaluated several for their usefulness for characterizing these communities and their potential
to assist in biofloc management protocols. These methods have been employed in both large outdoor tanks and greenhouse
raceways.
Traditional membrane dissolved oxygen sensors foul within hours of immersion in these biofloc systems and are of little use for
continuous monitoring and for operating automated aeration or emergency liquid oxygen systems. A YSI ® optical DO sensor
and a YSI ® pulsed polarographic DO sensor, both equipped with automated wipers, were mounted on the same YSI ® 6600 sonde
and deployed in a mature biofloc community for 52 consecutive days without maintenance. Both sensors performed equally
well and data obtained from them had considerably less random variability than readings conducted twice daily by a technician
using a handheld DO meter. Another sonde equipped with YSI ® phycocyanin and phycoerythrin Blue-green Algae sensors was
deployed in a raceway for 28 days to monitor cyanobacterial abundance. Although the system’s salinity was 35 ppt, the “freshwater”
phycocyanin sensor yielded data with less random noise and provided more accurate cyanobacterial estimates compared
to visual cell counts conducted with an epifluorescent inverted microscope equipped with cyanobacterial specific filters.
Several techniques were employed to better characterize the structure and activity of the microbial biofloc community. These
included direct microscopic observations using a scaled ranking system, and epifluorescent microscopy coupled with computer-aided
image analysis to quantify relative levels of algae, cyanobacteria, and heterotrophic bacteria. Tracking relative
bacterial abundance by quantification of fatty acids characteristic of bacteria (branched and odd chains) proved very effective.
Total community metabolism and suspended microbial community metabolism were measured in situ and in a specially designed
rotating incubator. Shrimp respiration was also estimated from lab respirometry. These techniques estimated that the
microbial biofloc community was responsible for approximately 80% of the total system oxygen demand. Critical evaluation
of these measurement techniques found some not to be useful, some appropriate in a research setting to develop protocols, and
some suitable for routine use by producers to improve system management.

INTENSIVE CULTIVATION OF Acartia tonsa (COPEPODA: CALANOIDA) OVER MULTIPLE
GENERATIONS DOES NOT NEGATIVELY AFFECT PRODUCTION
Jason T. Lemus and Rey de la Calzada
Thad Cochran Marine Aquaculture Center
The University of Southern Mississippi
Gulf Coast Research Laboratory
Ocean Springs, MS 39564 USA
jason.lemus@usm.edu
Copepod nauplii in the Order Calanoida are the optimum food for first feeding fish larvae; however, production from intensively
managed mass cultures is often inconsistent over multiple generations. We tested the hypothesis that culturing Acartia
tonsa (Order Calanoida) at intensive densities over several generations affects egg production, survival and size of copepods.
The investigation was initiated by stocking copepod eggs at 0.5/L, 1.0/L and 1.5/L in replicates of five. Adults developed from
eggs 9-11 days after stocking eggs and eggs produced by these adults were collected on days 12-16. On day 16 population survival
was determined and eggs from all replicates within each stocking density were combined and used to start a subsequent
generation of five replicates at the respective test densities. Eggs produced per day (egg production) and survival at day 16
(survival) were determined in the second, third, and fourth generations, egg diameter (egg size) and nauplii length (nauplii size)
were determined in the first and fifth generations, and adult cephalosome length (adult size) was determined in the sixth generation.
A general linear model was used to test for effects of generation (generation) and egg stocking density (stocking density)
on egg production and survival and for the effect of egg stocking density on copepod size. Bonferroni’s test was used to test for
significant differences among pair-wise comparisons. Student’s T-test was used to test for differences in egg and nauplius size
between the first and fifth generation. Significance level was set at 0.05 for all comparisons.
Egg production significantly increased with stocking density. Egg production was significantly greater in the third generation
than the first and second generations, which were not significantly different than each other. Survival was not affected by
stocking density. Survival in the fourth generation was significantly greater than the third generation, but not different than the
second generation which was not different than the third generation. Male and female adults were larger in the first than in the
sixth generation; however, egg szie was significantly greater in the fifth than the first generation. Nauplii size was marginally
greater in the fifth than the first generation (p=0.072). There was no effect of stocking density on egg, nauplii, and adult sizes.
The study results suggest that repeatedly culturing the same population of copepods in batches over multiple generations is
unlikely to depress the demographics. Inconsistencies with respect to size of copepods with respect to generation suggest that
inconsistent production of copepods cultured intensively is random error. We suggest that inconsistencies in production of
copepods managed for intensive mass production can be minimized by adherence to clearly defined management procedures
for the copepod production.
1

190
USE OF SETTLING TANK CLARIFIERS TO MANAGE THE MICROBIAL BIOFLOC
COMMUNITY IN SUPERINTENSIVE, ZERO-EXCHANGE SHRIMP PRODUCTION
SYSTEMS
Beth L. Lewis*, John W. Leffler, Andrew Ray, Heidi Atwood, Jason Haveman, Alisha Lawson, Brad McAbee,
Andrew Shuler, Jesus Venero and Craig L. Browdy
SCDNR Waddell Mariculture Center
211 Sawmill Creek Road
Bluffton, SC 29910 USA
ThomasBL@dnr.sc.gov
Superintensive, zero exchange, greenhouse-based systems offer many advantages for U.S. shrimp farmers including multiple
crops per year, reduced pollutant effluents, and reduction of harmful shrimp pathogens. Water quality is dependent upon the
rich microbial community composing and living on the abundant particulate matter that dominates these systems. In both
raceways and large outdoor tanks, studies at the Waddell Mariculture Center have used settling tank clarifiers to manipulate
this “biofloc” community. This “cropping” removes the nutrient and organic-rich biofloc particles from the water column and
they are collected as sludge in the clarifier until periodic removal. The settling tank clarifiers use an air-lift to pump water into
a settling chamber where it’s flow rate slows, allowing suspended solids to settle. Low particulate water returns to the shrimp
tank by gravity.
Weekly measurements demonstrated that the clarifiers were effective in reducing significant amounts of total suspended solids
(TSS). Since most of the biofloc is biologically formed within the system, volatile suspended solids (VSS) followed a very
similar pattern as TSS. Although TSS may be the most direct measure of clarifier effectiveness, it requires at least half a day to
obtain results. For routine management of the clarifiers, a simple field parameter is required for daily management decisions.
Turbidity is a complex parameter that is difficult to interpret biologically and may not work with all biofloc communities.
However we found it to be an effective management tool for operating the clarifiers on a routine basis. Research is currently
underway using a terrestrial and an underwater sensor to simultaneously measure photosynthetically active radiation (PAR)
and to report percent penetration and light extinction coefficient almost instantaneously. This appears to an easily applied,
biologically interpretable parameter for managing clarifiers.
Cropping significantly reduces suspended solids. In some cases autotrophy increases dramatically as measured by total
spectrophotometric chlorophyll and by epifluorescent microscopy with computer assisted image analysis. This probably
results from increased light penetration. In other cases, cropping had no significant effect on total chlorophyll concentrations,
primarily due to an abundance of Oosystis sp. dominating the biofloc. Based on several studies we have conducted, proper
biofloc management through cropping tends to increase shrimp growth and leads to reduced dissolved oxygen demand, reduced
dissolved nitrate, and reduced bacterial and cyanobacterial densities.

ANALYZING RESULTS THROUGH TIME USING A MULTIVARIATE APPROACH
Heidi A. Lewis* and Christopher C. Kohler
Fisheries and Illinois Aquaculture Center
Southern Illinois University
Carbondale, IL 62901-6511 USA
hal7e7@siu.edu
Aquaculture experimental design commonly involves measurement of multiple dependent variables in response to one or more
experimental treatment(s). Frequently, repeated measurements are taken over the course of the experiment to monitor progress.
Often these intermediary measurements are removed from the data set and the final measurement is used to evaluate the
predetermined hypothesis. Utilizing a hypothesis testing procedure that includes all data collected over the course of the trial
can provide researchers with the ability to interpret the rate of change as well as the final effect of the experimental treatment.
However, analyzing each dependent variable separately at each time point using univariate analysis of variance (ANOVA) will
inflate the risk of committing a Type I error. Whereas, multivariate analysis of variance (MANOVA) approach to repeated
measures allows researchers to statistically analyze multiple dependent variables simultaneously, therefore lowering the incidence
of Type I error and generating a more valid p-value. This multivariate approach to repeated measures uses MANOVA to
test within subjects hypotheses of flatness and parallelism within the data set. Further univariate hypothesis testing evaluates
between subject factors using one-way ANOVA. Advantages and disadvantages of utilizing profile analyses in aquaculture
nutrition research will be discussed with emphasis on juvenile and broodstock research.
1 1

1 2
SUNSHINE BASS EMBRYONIC AND LARVAL SURVIVAL DIFFER IN RESPONSE TO
REMOVAL OF MARINE OIL FROM MATERNAL BROODSTOCK DIETS
Heidi A. Lewis, Jesse T. Trushenski, Ryan L. Lane and Christopher C. Kohler
Fisheries and Illinois Aquaculture Center
Southern Illinois University
Carbondale, IL 62901-6511 USA
hal7e7@siu.edu
Development of suitable broodstock feeds while reducing dependence on marine oil resources is limited by not knowing fatty
acid requirements for many taxa. Quantification of fatty acid utilization by larvae during the endogenous feeding period will
provide information on essentiality of specific fatty acids for larval development. Accordingly, we monitored sunshine bass
larval survival and fatty acid profile change throughout the endogenous feeding period after altering oocyte fatty acid composition
by feeding female white bass broodstock different dietary lipid sources.
Female white bass (790 ± 19.7 g; mean ± standard error) were stocked into a recirculation system, 7 fish per tank. Triplicate
tanks were fed diets (45% protein and 14% crude lipid) containing graded levels (0, 33, 67, or 100%) of flax to menhaden fish
oil as the primary dietary lipid source. All fish were fed daily to apparent satiation for 6 months. Oocytes were collected by
manual stripping and fertilized using striped bass semen. Egg viability and fertilization were assessed, and larval survival and
fatty acid profile were monitored through 5 days post hatch (DPH).
The fatty acid composition of the oocyte largely reflected maternal dietary intake. However, at the end of the endogenous
feeding period, larval composition deviated from initial oocyte composition (Table 1). Embryonic survival of 100% fish oilfed
broodstock (20.7 ± 3.5%) was significantly greater than 100% flax oil-fed broodstock (6.6 ± 3.8%). Alternatively, larval
survival was significantly reduced in progeny of the 100% fish oil-fed broodstock (51.6 ± 4.7%) compared to those fed the
100% flax oil diet (72.0 ± 5.1%). Differential responses in embryonic and larval survival resulted in comparable total larval
yields at 5 DPH among all maternal dietary treatments (11,000 ± 4,200 female -1 ).
These data support the use of flax oil as a suitable lipid source for marine oil replacement in white bass broodstock diets.
Although n-3 HUFA from fish oil is essential for embryonic survival, sunshine bass appear to have lower n-3 HUFA requirements
after hatch. Larval survival was highly dependent on the presence of 18-carbon polyunsaturated fatty acids (18-C PUFA)
which were present due to flax oil inclusion in maternal diets. Further research is needed to maximize both embryonic and
larval survival of sunshine bass through provision of both dietary 18-C PUFA and n-3 HUFA to maternal white bass broodstock.

SLOPE RATIO ANALYSIS IS A ROBUST AND PRECISE WAY TO DETERMINE RELATIVE
BIOAVAILABILITY OF NUTRIENTS FOR AQUACULTURED ANIMALS
Peng Li* and Delbert M. Gatlin III
Department of Animal Science
Texas A&M University
College Station, Texas 77843-2471 USA
lipeng@tamu.edu
Nutrient bioavailability estimation is important, not only for screening ingredients that provide the most cost-effective nutrients,
but also for developing microencapsulation/coating processes to increase retention of certain nutrients and overall utilization
efficiency of aquacultured animals. Traditionally, nutrient bioavailability is obtained either by comparing the minimum requirement
for a specific nutrient in different forms established by broke-line regression or polynomial regression, or by comparing
response indices such as weight gain and feed efficiency of animals fed the nutrient in different forms. Bioavailability of nutrients
can be most precisely estimated by slope-ratio assay. In this assay, graded nutrient intake levels are created by varying the
dietary inclusion level of a particular nutrient or ingredient. The response, such as whole-body nutrient concentration and retention
by animals fed the test ingredient is related to nutrient intake, and the slope of the regression line is compared with that from
animals fed the nutrient from a different source. Our laboratory has used slope ratio analysis to define relative bioavailability
of DL-met vs. methionine hydroxy analog for hybrid striped bass (Figure 1 and 2), seleno-methionine vs. sodium selenite for
hybrid striped bass (Figure 3) and α-tocopheryl acetate vs. α-tocopherol succinate for red drum (Figure 4). We observed the
slope ratio analysis is a very robust and precise method in bioavailability estimation in aquaculture nutrition.
1 3

1 4
LIPID AND FATTY ACID REQUIREMENTS OF TILAPIA
Chhorn Lim and Mediha Yildirim-Aksoy
Aquatic Animal Health Research Laboratory
USDA-ARS, MSA
Auburn, AL USA
Chhorn.lim@ars.usda.gov
Dietary lipids are an important source of highly digestible energy and are the only source of essential fatty acids required for
normal growth and development. They are also carriers and assist in the absorption of fat-soluble nutrients, such as sterols and
fat-soluble vitamins, serve as a source of phospholipids and participate in the synthesis of hormones, prostaglandins and other
metabolically active compounds. Tilapia have been reported to have a dietary requirement for linoleic (n-6) series of fatty acids.
The optimum dietary levels of n-6 (18:2 n-6 or 20: 4 n-6) have been estimated to be 0.5 and 1% for redbelly tilapia (Tilapia
zillii) and Nile tilapia (Oreochromis niloticus), respectively. This information for other species has not been determined, but it
has been suggested that blue tilapia (O. aureus) have a relatively high requirement for n-6 fatty acid. Tilapia may also have a
requirement for linolenic (n-3) series of fatty acids. However, the optimum dietary requirement level for n-3 has not been determined.
The presence of high levels of either n-6 or n-3 may spare the requirement of the other, although n-6 appears to have
better growth promoting effect than n-3. Tilapia appear to possess the ability to desaturate and chain elongate 18:2 n-6 to 20:4
n-6 and 18:3 n-3 to 20:5 n-3 and 22:6 n-3.
Research evaluating various sources of dietary lipids showed that plant oils rich in n-6 and/or n-3 series of fatty acids, such as
soybean oil, corn oil, sunflower oil, palm oil products and linseed oil are good lipid sources for tilapia. Beef tallow is a poorly
utilized by tilapia when used as the sole lipid source. However, it can be used in combination with other lipid sources providing
that their essential fatty acid requirements are met. Information on the nutritional value of fish oil for tilapia is inconsistent.
Some studies have shown that the nutritional value of fish oil is similar to that of plant oils, while others reported poor performance
of fish oil-containing diets. Fish oil provided good spawning performance for tilapia broodstock reared in saline water,
while in freshwater, good reproductive performance was obtained with plant oil (soybean oil). The sparing effect of dietary
lipids on protein utilization has also been demonstrated. However, tilapia do not tolerate as high a dietary lipid level as do salmonids.
A dietary lipid level of 5 to 12% has been suggested to be optimum for tilapia.
The discrepancy between the results of various studies could be due to various factors such as species, fish sex and size, previous
dietary history, source and levels of dietary lipid, composition and nutrient content of experimental diets, feeding duration
and practices, and environmental conditions (salinity, temperature, etc.).

THE USE OF HYDROGEN PEROXIDE (H 2 O 2 ) IN CULTURE SHRIMP Litopenaeus vannamei
(BOONE, 1931)
João Paulo Viana de Lima; Weruska Melo Costa, Luis Otavio Brito; Wanessa de Melo Costa, Alfredo Olivera Gálve
Pernambuco Federal Rural University
Fisheries and Aquaculture Department
Dois Irmãos, 52171-900
Recife, Pernambuco, Brazil
engpescalo@hotmail.com
The use of hydrogen peroxide is a source of oxygen for the cultivation of aquatic organisms capable of increasing concentrations
of the gas, making it in the water column so that can be used by aquatic organisms to the metabolic processes.
The objective was to evaluate the use of peroxide in the system of cultivation of shrimp. The experiment was conducted in
three stages.
The first stage was in 350L tank, the second was in tanks, and the third was 40.000L in nurseries of 20.900m3.
Showed results were submitted to ANOVA, then to Tukey test.
The results showed that there were significant differences. In phase 1 the best results of production of shrimp were found with
the treatments C (3 ppm) and D (5 ppm).
Regarding the increase in the concentration of dissolved oxygen in water, application of hydrogen peroxide both in phase 2 and
3 demonstrated efficient.
In relation to the temperature and pH there was no significant difference.
1

1
RELATIONSHIPS AMONG THERMAL STRESS, BLEACHING AND OXIDATIVE DAMAGE
DURING THE CULTURE OF HERMATYPIC CORAL, Pocillopora capitata
Marco A. Liñán-Cabello, Laura A. Flores-Ramírez and Alfredo Mena-Herrera
Acuacultura/Biotecnología, FACIMAR
Universidad de Colima
Km 19.5 carretera Manzanillo-Barra de Navidad Manzanillo
Colima, México
linanmarco@hotmail.com
Corals have a symbiotic relationship with the photosynthetic zooxanthellae that live within their endodermal cells, providing
energy to the coral tissues. Under high temperature conditions, the stability of this symbiosis is adversely affected, leading to
bleaching in a wide variety of coral species. Various in vitro studies have shown that symbiotic dinoflagellates maintained in
culture are susceptible to thermal stress.
To examine the response to exposure to a thermal gradient in
coral in vitro, we assessed the effect of a gradual 10 °C temperature
increase (22 to 32 °C over 10 h) on normal (N), partially
bleached (P) and control (C) samples collected from different
branches of the same coral (Pocillopora capitata). We
examined markers of oxidative stress, including lipid peroxidation
(MDA) and superoxide dismutase (SOD) (Tab.1) activity,
indicators of bleaching, including chlorophyll a (Chl a) and carotenoid
pigment (PC) levels, as well as zooxanthellae density
(Fig. 1). Our results revealed that N, P and C coral samples all
contained higher levels of PC versus Chl a. The levels of both
pigments increased as the temperature increased from 22 to 28
°C only in N and C samples, whereas P samples showed less
cellular damage than N and C samples at temperatures between
26 and 28 °C, and had greater antioxidant activities at temperatures
between 26 and 30 °C. The rate of zooxanthellar expulsion
consistently increased with temperature in all three coral
types across the entire temperature range.
We found that partially bleached samples (P) of P. capitata
tolerated short-term stress (at temperatures up to 28 °C) more
effectively than did normal (N) or control (C) samples of the
same coral. This P sample tolerance could be a consequence of
its continuous exposure to a regime of short duration thermal
variation (4–6 °C).
Collectively, these results indicate that temperature has a direct
effect on the antagonistic relationship between temperature-
induced damage and protective antioxidant mechanisms in this
type of coral.

DETECTION OF YELLOW HEAD DISEASE IN INTENSIVE FRESHWATER PRODUCTION
SYSTEMS OF Litopenaeus vannamei
Liñán-Cabello Marco A., Sánchez-Barajas Maximiliano and Mena-Herrera Alfredo
Facultad de Ciencias Marinas
Universidad de Colima
Km. 19.5 Carretera Manzanillo-Barra de Navidad
Manzanillo Colima México CP 28860
linanmarco@hotmail.com
The culture of penaeid shrimps in inland well waters with low salinity represents an important development in aquaculture.
Yellow head disease of shrimps has been reported in many countries in Asia and Latin America, but, in most cases, the etiologic
agents have not been clearly defined. In the present study, we used reverse transcription-nested polymerase chain reaction (RT-n
PCR) to detect yellow head virus (YHV), white spot syndrome virus (WSSV), and gill-associated virus (GAV), during single
culture cycles of Litopenaeus vannamei on two farms (F1 and F2), located in Colima, Mexico.
The shrimps were intensively cultured in freshwater, with densities of 70 organisms org m -2 and 25 org m -2 and hardness values
of 390 and 450 mgL-1 (as CaCo3), respectively. The average salinities throughout the year were 0 ppm. The results obtained
indicated that the samples were negative to WSSV and GAV. YHV was present on each farm, with a prevalence of 13%, over
60 d of observation. During this time, the growth of shrimps decreased on F1. The infections obeyed the principles of horizontal
transmission, and we discuss various factors, such as increases in shrimp density, and consequent lower levels of oxygen, that
could increase the effects of YHV on the health of shrimps on farm F1. We consider that the observed infection obeyed the
principles of horizontal transmission. It is important to determine if the YHV we detected is a product of dissemination from
other farming areas, or if it corresponds to an endemic variety. If the latter, our virus would be considered a member of the
YHV complex.
1

1
ACOUSTIC CONDITIONING AND OCEAN RANCHING OF BLACK SEA BASS Centropristis
striata NEAR WOODS HOLE, MASSACHUSETTS
Scott Lindell*, Simon V. Miner, Clifford A. Goudey, Steve Page, Hauke Kite-Powell and John Murt
Scientific Aquaculture Program
Marine Biological Laboratory
7 MBL St.
Woods Hole, MA 02543 USA
slindell@mbl.edu
Acoustic ranching consists of training fish to school to an area via a sound stimulus that is coupled with a food reward (classical
Pavlovian conditioning). It may present a novel opportunity to raise fish with less environmental impact and at less expense than
typical open-ocean fish farms. Some advantages include (i) lower capital costs to construct and install a feeding and recapture
station (versus a cage engineered to withstand open ocean conditions), (ii) lower operating costs associated with maintaining
a feeding station, (iii) lower feed costs because fish have opportunities to forage on wild food as well as formulated diets, (iv)
lower potential impact on the environment due to natural dispersion of fish and their wastes, and (v) the technology could aid
stock replenishment efforts by gently weaning hatchery-raised fish from pelletted diets to fending for themselves in the wild.
This project presents the first attempt to farm marine fish with acoustic ranching in North America. Black sea bass (Centropristis
striataa) are a popular commercial and recreational fishery that has declined significantly in past decades. We purchased
7,000 one gram black sea bass juveniles from Great Bay Aquaculture in September 2007. In June 2008 we erected and installed
an AquaDome TM , a 10 m (32’) wide by 5 m (16’ high geodesic dome covered with a 1-inch, plastic-coated wire mesh, on the
sandy seafloor in Buzzards Bay near Woods Hole Massachusetts. The site was selected for its proximity to a natural rocky
outcropping that makes ideal juvenile fish habitat. The AquaDome was fitted with a 7.5 cm (3”) feeding tube, an underwater
speaker and underwater cameras to monitor and record fish behavior at sound generation and feeding events. Approximately
5,000 tagged black sea bass were stocked into the AquaDome in July 2008. The fish were trained in the cage by feeding them
twice a day in tandem with a sound cue (a 320 Hz tone) (a 320 Hz tone). Once the training was completed, some of the one-inch
mesh on the AquaDome was replaced with 10 cm (4”) mesh so that the fish could swim out and set up residency on the nearby
rocky bottom. The fish were able to swim back in during sound cue and feeding events.
Based on laboratory experiments we conducted with juvenile black sea bass over the winter and spring of 2008, it takes about
4 weeks to properly train a group of fish to consistently respond to a sound cue and move to a prescribed area in anticipation
of receiving a food reward. We have also documented that black sea bass are very elastic in their diet and can switch between
eating pellets and wild forage quite readily. Furthermore, we have demonstrated that once the learned behavior of responding
to the sound cue has been mastered it is can be consolidated in the fish’s memory for at least 4 weeks.
We will present results of our laboratory and field trials and our efforts to recapture the majority of these fish, evaluate their
growth and measure the potential economic advantages of this method of fish production.

EFFECTS OF STANDARD AND HIGH-FAT DIETS WITH OR WITHOUT A DAIRY/YEAST
PREBIOTIC ON PERFORMANCE OF GOLDFISH Carassius auratus IN OUTDOOR POOLS
Harold Phillips, Nick Kinsey and Rebecca Lochmann*
Aquaculture Research Center
University of Arkansas at Pine Bluff
Pine Bluff, AR, 71601 USA
rlochmann@uaex.edu
Previous studies with high-fat diets or prebiotics have shown beneficial effects in baitfish. In this study, we determined the responses
of goldfish in outdoor pools fed diets with standard (4%) or high (10 %) amounts of added lipid either with or without
a dairy/yeast prebiotic. Four diets similar to a commercial diet (30% protein and 9.6 kg energy/gram of protein) were formulated.
Two diets contained the same protein components (primarily fish and poultry meals) and differed only in the amount of
added lipid (4 or 10% poultry fat). The diet with 4% fat was the control. Two other diets were similar to diets 1 & 2 except they
contained 2% prebiotic.
Four hundred (0.36 + 0.002g in individual mass) fish were randomly stocked into each of four plastic-lined 4.1-m 3 pools
containing reservoir water and maintained static during the study. Fish are being fed twice daily to apparent satiation and
group-weighed every 2 wks to track growth. Chlorophyll a and Secchi depth are being measured to assess natural foods. At
four weeks, there were no differences (P>0.05) in mean individual weight gain (0.66+0.03g) among treatments. The feeding
trial will be conducted a minimum of 8 weeks, then final production data will be taken and fish will be subjected to bacterial
challenge. Results of this data will be compared to those of a concurrent trial being conducted in indoor aquaria to determine
the efficacy of these diets in the presence and absence of natural foods.
1

200
INTERACTIVE EFFECTS OF VITAMINS C AND E ON THE GROWTH AND HEALTH OF
LARGEMOUTH BASS Micropterus salmoides
Daryl Weldon*, Rebecca Lochmann, Ruguang Chen, Andrew Goodwin and Steve Rawles
Aquaculture Research Center
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601 USA
dweldon@uaex.edu
The objective of this study was to determine the interactive effects of ascorbic acid (AA) and α-tocopherol (α-T) on growth,
survival, hematological responses, and non-specific immune responses (lysozyme and alternative complement activity) of juvenile
largemouth bass cultured at 26 °C. Experimental fish were acclimated and maintained on the basal diet deficient in both
AA and α-T for two weeks prior to stocking to deplete stores of both vitamins. Ten fish with an initial weight of 50.2 ± 0.11g
(mean+SD) were stocked into each of eighteen 150-L isoform tanks configured in a recirculation system having a flow-rate of
2L/min. and maintained with dechlorinated municipal water. Tanks were randomly assigned in triplicate to one of six dietary
treatments supplemented with 0, 100, or 250 mg of AA in the form of ascorbyl-2-monophosphate and either 0 or 50 mg of α-T
acetate powder per kg of diet in a 3 x 2 factorial design. The six isocaloric semipurified diets contain approximately 12% lipid,
880.3 kJ of energy, 42% protein, 13.7% ash, and 80% dry matter.
Fish are being fed to apparent satiation once daily and excess feed is removed, dried, and subtracted from the amount of feed
offered to estimate feed intake. Fish are weighed every two weeks to monitor weight gain and feed conversion ratio (FCR).
Temperature and mortalities are recorded daily and water quality parameters are monitored and recorded weekly. The fish were
treated with antibiotics on four occasions for systemic Columnaris infections. Water hardness and alkalinity were maintained
at approximately 45 mg/L and salinity at 2 g/L to discourage Columnaris outbreaks. For the rest of the trial a total hardness and
alkalinity of approximately 100 mg/L and a salinity

EFFECTS OF STANDARD AND HIGH-FAT DIETS WITH OR WITHOUT A DAIRY/YEAST
PREBIOTIC ON PERFORMANCE OF GOLDFISH Carassius auratus IN AQUARIA
Ruguang Chen, Harold Phillips and Rebecca Lochmann*
Aquaculture Research Center
University of Arkansas at Pine Bluff
Pine Bluff, AR, 71601 USA
rlochmann@uaex.edu
Diets that enhance stress resistance and prolong survival of goldfish would enhance the profitability of bait and feeder fish
production. We conducted a feeding trial with goldfish in aquaria to determine whether practical diets supplemented with a
dairy-yeast prebiotic , and 4- or 10% supplemental lipid could improve growth, survival, feed conversion, body composition,
or survival after bacterial challenge. Four diets similar to a commercial diet (30% protein and 9.6 kg energy/gram of protein)
were formulated. Two diets contained the same protein components (primarily fish and poultry meals) and differed only in the
amount of added lipid (4 or 10% poultry fat). The diet with 4% fat was the control. Two other diets were similar to diets 1 & 2
except they contained 2% prebiotic. Thirty fish (0.57+0.002 g in individual mass) were stocked into each of four replicate 110-
L tanks per treatment in a recirculating system with municipal water supplemented with calcium chloride to maintain hardness
above 50 mg/L, and salt at 2 mg/L to enhance survival. Fish are being fed twice daily to apparent satiation and group-weighed
every 2 weeks to track growth. Weight gain (0.89+0.02 g) and survival (96.3+0.7 %) did not differ at 4 weeks. The feeding
trial will be conducted a minimum of 8 weeks, then final production data will be taken and fish will be subjected to bacterial
challenge. Results of this data will be compared to those of a concurrent trial being conducted in outdoor pools to determine the
efficacy of these diets in the presence and absence of natural foods.
201

202
EFFECTS OF DIETARY PROTEIN SOURCE AND INTERACTIVE EFFECTS OF PROTEIN
AND LIPID SOURCE ON CHANNEL CATFISH EGG BIOCHEMICAL COMPOSITION, EGG
AND FRY PRODUCTION, AND EGG AND FRY QUALITY
Todd D. Sink* and Rebecca T. Lochmann
University of Arkansas at Pine Bluff
1200 N. University Drive
Pine Bluff, AR 71601 USA
tsink1@uaex.edu
Four diets (36% protein, 10% supplemental lipid) differing in protein source or protein and lipid source were used to examine
the effects of protein source and protein lipid source interactions on egg and fry production in channel catfish. Diets included
(1; control) fish meal, poultry by-product meal, and 10% menhaden fish oil (FM-PBM-FO), (2) fish meal, poultry by-product
meal, 5% menhaden fish oil, and 5% poultry fat (FM-PBM-FO-PF), (3) poultry by-product meal and 10% menhaden fish oil
(PBM-FO), or (4) all plant protein sources and 10% menhaden fish oil (PP-FO). Four-year-old broodfish were stocked at a ratio
of 3 females: 1 male into each of 16 tanks (2.5 m, 2,365 L) in February of 2008 and fed assigned diets up to 5 times weekly
throughout the study dependant upon water temperatures. Spawning cans were placed in the tanks in April and checked three
times weekly through the second week in July.
Twenty-five of 48 possible spawns were collected
(52.1% spawn success). There were no differences
in brood mortality among the diet treatments. Fish
fed the FM-PBM-FO-PF diet had greater spawning
success, fecundity, egg mass weight, and hatch
rates than fish fed PP-FO, but most parameters
were not different from the control or PBM-FO
diet. No significant protein by lipid source interaction
was found for any parameter between the
control and FM-PBM-FO-PF diets, indicating that
poultry fat can be used to partially replace fish oil
in catfish broodstock diets when a variety of animal
and plant protein sources are used. Analysis of
protein source revealed that fish fed diets containing
FM and PBM (in addition to plant proteins)
produced more eggs than fish fed only PBM or PP.
Additional analyses of eggs and fry are ongoing,
but initial data suggests that catfish broodstock
produce more eggs when fed a variety of animal
protein and lipid sources.
Measurement
parameter Diet
FM-PBM-FO FM-PBM-FO-PF PBM-FO PP-FO
Total broodstock
survival (%) 81.3 75.0 81.3 87.5
Female broodstock
survival (%) 66.7 50.0 66.7 83.3
Spawning success
(%) 58.3 a 58.3 a 58.3 a 33.3 b
Fecundity (eggs·kg
female -1) 6,432 a 6,420 a 4,060 a,b 1,072 b
Spawn time (days
post can insertion) 25.3 23.7 31.0 43.0
Water temperature
at spawning (°C) 26.0 26.7 26.2 27.7
Egg mass weight,
matrix intact (kg) 0.722 a,b 0.814 a 0.436 a,b 0.357 b
Total egg volume,
matrix removed
(mL) 847 a,b 1019 a 540 a,b 490 b
Individual egg
weight (g) 0.0298 a,b 0.0338 a 0.0281 b,c 0.0240 c
Eggs·mL -1 23.1 a 21.2 a,b 21.6 a,b 16.3 b
Eggs·spawn -1
(volumetric) 19,654 a,b 21,814 a 11,782 a,b 7,991 b
Eggs·spawn -1
(mass) 20,268 a 19,844 a 11,132 a,b 7,458 b

OPTIMIZING FEEDING STRATEGIES FOR TANK CULTURE OF SUNSHINE BASS USING
MICROCYST Artemia NAUPLII AS AN INTERMEDIATE STEP BETWEEN ROTIFERS AND
STANDARD Artemia NAUPLII
Steve E. Lochmann* and Gerald M. Ludwig
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
1200 N. University Dr., Mail Slot 4912
Pine Bluff, AR 71601 USA
slochmann@uaex.edu
Tank culture of sunshine bass larvae has typically required rotifers at the time of first feeding. Recently, sunshine bass larvae
have been cultured without rotifers, by using Artemia nauplii from microcysts at first feeding. Survival rates of 43% have
been reported culturing sunshine bass larvae on microcyst Artemia nauplii alone. We wanted to see if survival and growth of
sunshine bass larvae would be improved by including rotifers (Brachionus plicatilis), microcyst Artemia nauplii, and standard
Artemia nauplii in sequence during a production run. This experiment was comprised of three treatments with three replicates
per treatment. Sunshine bass larvae, 4 dph (4.0 + 0.1 mm SL), were stocked into 100-L recirculating tanks at a rate of 75 larvae/
L. The first feeding treatment was rotifers (40/mL) followed by standard Artemia nauplii (8/mL). The second feeding treatment
was microcyst Artemia nauplii (20/mL) followed by standard Artemia nauplii (8/mL). The third feeding treatment was
rotifers (40/mL) followed by microcyst Artemia nauplii (4/mL) followed by standard Artemia nauplii (8/mL). By day 8, all of
the feeding treatments were fed standard Artemia nauplii only. Water quality (DO, pH, TAN, and hardness) of the recirculating
systems was tested daily. Fingerlings were harvested on day 21 of the experiment. Survival was determined gravimetrically.
Thirty larvae from each tank were photographed at the end of the study. Standard lengths of larvae were determined using image
analysis software. We compared survival and growth among treatments using single factor analyses of variance.
Average (SD) individual standard length of larvae was 9.08 (1.43) mm and ranged from 6.46 to 15.05 mm. Average length
of larvae did not vary among treatments (F=0.81, df=2, P=0.489). Variability in survival among tanks was high (Table 1).
Survival ranged from 9% to 96%, and varied somewhat among treatments (F=3.41, df=2, P=0.102). The lowest two survival
rates (9% and 12%) occurred in treatment two and the highest two survival rates (70% and 96%) occurred in treatment three.
It appears that matching food size to larval size, as larvae grow, is likely to result in better survival. However, growth of larvae
does not appear to improve similarly.
Table 1. Survival and growth of sunshine
bass for three different feeding strategies.
Treatment
1 2 3
Survival (%) 31 23 68
Length (mm) 9.1 9.6 8.5
203

204
THE USE OF FOURIER TRANSFORM INFRARED SPECTROSCOPY TO PREDICT
OVARIAN ATRESIAAND CAVIAR QUALITY IN WHITE STURGEON (Acipenser
transmontanus)
Xiaonan Lu and Barbara Rasco
School of Food Science
Washington State University
Pullman, WA 99163 USA
xiaonan_lu@wsu.edu
Fourier Transform Infrared Spectroscopy (FT-IR, 4000-400 cm -1 ) spectral features of white sturgeon (Acipenser transmontanus)
(N=11) plasma and roe were determined over a nine month period to predict when fish entered atresia. The objective was
to determine whether changes in spectral features could be used to predict caviar quality. FT-IR spectra of plasma changed at
various maturity levels. Differences in plasma biochemical composition in the sex steroids region (around 3000 cm -1 ) were
detected along with changes in the concentration of plasma vitellogenin (around 1080 cm -1 ). By using Principal Component
Analysis (PCA), it was possible to clearly segregate fish by maturity stage due to differences in spectral features of the plasma.
The stage of maturity (late vitellogenesis vs. early atresia) could be predicted ~ 70% of the time using Soft Independent
Modeling of Class Analogy (SIMCA) models. Also, a rigorous Partial Least Squares (PLS) model was established to predict
Polarization Index (PI) values between 0.1 and 0.3 (R=0.98, SEP=1.01%) based upon differences in plasma spectral features
(Fig.1). FT-IR spectra of fish plasma may provide a useful tool for assessing sexual maturity in female sturgeon. Few changes
were observed in spectral features of the roe recovered during late vitellogenesis or early atresia. This technique may reduce
the need for traditional invasive and stressful surgical biopsy methods for PI determination currently used to predict the timing
of roe harvest.
Figure 1. Comparison between actual and predicted PI values
(range: 0.1 and 0.3) for sturgeon females

ISOLATION, IDENTIFICATION AND BIOLOGICAL FUNCTION OF A PUTATIVE SHRIMP
β-Actin Promoter from Litopenaus Vannamei
Yuanan Lu, Marcus Soderlund and Chengxiang Wu
Department of Public Health Sciences
University of Hawaii
Honolulu, Hawaii 96822 USA
ylu@pbrc.hawaii.edu
Shrimp farming has become an establishing industry in many countries over the world. However, viral diseases have proven
to be a serious threat to shrimp farming and pathogenic viruses are frequently reported to cause 100% shrimp mortality and
tremendous economic losses worldwide. There are currently no effective methods to treat viral diseases. The use of molecular
biology techniques to produce pathogen resistant strains of shrimp through genetic transformation technology is considered a
highly promising strategy for control of shrimp viral diseases. A major challenge to current development of such biotechnology
is the lack of a suitable host-derived promoter to drive the effective expression of desirable genes in shrimp. To meet this urgent
need for the development of transgenic shrimp, we have recently employed molecular cloning and sequencing techniques
and isolated and identified a putative promoter gene from litopenaeus shrimp (L. vannamei), shrimp β-actin promoter. This
represents the first promoter gene identified and reported from crustacean species. Biological function of newly discovered
shrimp promoter has been demonstrated using a newly constructed gene expression cassette system under the control of such
shrimp promoter. Experimental tests have confirmed high level of stable expression of red fluorescent protein (RFP), a reporter
transgene, and also an antiviral gene of shrimp white spot syndrome virus in a variety of in vitro cell cultures derived from both
mammalian and marine animal species using fluorescent microscopy, Western blot and enzyme-linked immunosorbent assay.
With the previously established jetPEI/DNA technique for gene delivery into the shrimp zygotes, this putative shrimp promoter
forms the essential base for genetic shrimp transformation technology and makes it possible for the development and production
of viral-resistant transgenic shrimp strains for world shrimp aquaculture.
20

20
THE USE OF AN INORGANIC AND ORGANIC ACID-BASED PRESERVATIVE AS STORAGE
CONTROL FOR FISH-OFFAL
Friedrich Weißbach and Christian Lückstädt*
ADDCON Nordic AS
3908 Porsgrunn, Norway
christian.lueckstaedt@addcon.net
Acid preservation of fish and fish viscera to produce fish silage is common practice. The final product has been widely used
in fish feed and as a feed component for fur animals. In many European countries it is a commonly used method of preserving
fish-by-products, using formic acid or its salts to keep the fish silage stable. However, due to regulatory restriction and
the limited tolerance to formic acid in fur animals such as mink and foxes, the accepted practice of preparing fish silage with
formic acid treatment is not always possible. The objective of the present study was therefore to investigate the effect of a new
chemical preservation method which excludes the use of formic acid.
The efficacy of a liquid blend of sodium disulphite, hexamethylenetetramine and sodium benzoate was evaluated when prepared
in aqueous solution by a special procedure (FURMIX, patent pending). This preservative was added to nearly 10 tonnes
of roughly minced Baltic herring (Clupea harengus) which was previously acidified by adding 13 litres of concentrated sulphuric
acid per tonne. The product blend was applied at a rate of 20 litres per tonne. The storage trial was carried out under European
summer conditions (approximately 20°C) in an open container and lasted for 3 months. The following parameters were
tested after one month and at the end of the trial (Table 1): bacterial counts, mould count, pH and total volatile nitrogen (TVN).
TVN is often used as an indication of the freshness of fish raw material. The main constituents of TVN are trimethylamine and
ammonia. Trimethylamine originates from bacterial decomposition and its presence in raw fish is thus taken as an indication of
bacterial growth, while ammonia arises from the breakdown of amino acids – thus reducing the quality of the available protein.
Levels of around 75 mg TVN per 100 g fish mass are regarded by the industry as limits for good quality fish silage.
The results clearly indicate that due to the addition of sulphuric acid and the unique acid-salt blend, the fish silage derived from
herring can be kept stable and in good quality for at least three months for further feed processes.

MODELLING OF NUTRITIONAL REQUIREMENTS IN AQUATIC SPECIES
Ingrid Lupatsch
Centre for Sustainable Aquaculture Research
Swansea University
Singleton Park
Swansea SA2 8PP, UK
i.lupatsch@swansea.ac.uk
Modelling of nutritional requirements is a mathematical setup used to predict and improve growth performance and feed efficiency
of the target species. A suitable model is ideally based on mathematical relationships that should be as simple as possible
but be able to supply reliable data for practical application.
In classical nutrition the requirements are separated into two components, first the amount of energy and protein needed to
maintain zero balance and the other component the energy and protein needed for growth above maintenance. Thus total energy
and protein requirements are considered the sum of maintenance plus growth. The requirement for maintenance is mainly a
function of the size of the fish and water temperature, and is proportional to the metabolic body weight. The requirement for
growth is dependent on the amount and the composition of the weight gain, including the energetic cost to deposit the new
growth.
The following equation specifies the formal approach to those calculations:
Requirement = a × BW (kg) b + c × gain
(kg) b : Metabolic body weight
a is the constant for given conditions (activity, temperature) expressed in kJ per unit of metabolic weight and characteristic of
a fish species
b is the exponent of the metabolic body weight and converts absolute weight to metabolic weight correcting for the decrease in
metabolic rate per unit of body weight as fish grow.
c - cost in units of energy or protein to deposit new growth.
Mathematical equations based on empirical studies will be presented to derive the necessary parameters to define:
• Maintenance requirements for energy and protein at different water temperatures
• Growth data – a description of the potential weight gain along the growth cycle at different temperatures.
• Feed intake - a prediction of the voluntary feed intake at increasing fish sizes and water temperatures.
• Description of protein and energy content with increasing fish size.
• Efficiency of utilization of dietary energy and protein to deposit energy and protein as growth.
Feeds can then be formulated and feeding tables established which are based on daily requirements for energy and protein
dependent on anticipated growth of the fish species in question.
20

20
THE EFFECT OF TWO DIFFENET SALINITIES ON CYST BIOMETRY OF Artemia urmiana
FROM URMIA LAKE REGIONS
Naseh Abdolah Zadeh, Samad Zare , Ramin Manaffar* and Alireza Asem
Department of Biology, Faculty of Science
Artemia and Aquatic Animals Research Institute
Urmia University
P.O.Box 165
Daneshkadeh Ave.
Urmia, Iran
Raminmanaffar@yahoo.com
Urmia Lake is one of the widest saline Lakes on world. It has no out let and restricted to almost 4000 km 2 area in North West
of Iran. A case way from the west to the east of the Lake now divided the Lake to two regions as North and West. Because of
its salinity (200-300 ppt) Artemia is solely animal can tolerate this habitat for many years in the Lake and is the most famous
animal in Aquaculture. Urmia Lake area by its wide rage of ecological conditions is known to have some Artemia populations
that already were demonstrated in many papers using by molecular and morphological tools. These populations can produce
cysts with different average size which is very important not only in aquaculture but also in marketing. To achievement the
cause of this variability in cyst diameter, which is genetically or environmentally, in this project the cysts of Artemia were
collected from 5 different regions of Urmia Lake from North and South. The cysts were hatched in optimum condition of hatching
procedure. The hatched nauplii reared in 1 l conical tubes for 20 day by feeding of unicellular algae and Yeast according to
optimum culture condition of Artemia in two different salinity of 75 and 150 ppt. 32 couple of adult Artemia from each populations
(Artemia urmiana, bisexual) were carried out to the 50 ml falcon tubes (a tube for each couple) in corresponded salinity.
The produced cysts from each population were collected every day of inside of falcon tubes by 150 micrometer sieves, for a
period of 90 days. Finally the samples of cyst were hydrated in D&K solution for a period of 16 hour and 150 individual of
each population were measured in diameter. The results and its statistic analysis in comparing with the original cyst collected
from the Lake in table 1.
Result and its statistically analysis revealed that, as well as previously released reports, the size of cyst in Urmia Lake is depend
to culture conditions approximately, but according to the results of this study in some stations the salinity conditions not be
able to influence the size of cysts comparing with original Lake cyst, in which shows the variability of cyst size is genetically,
partially. In conclusion for farm culture of Artemia urmiana for cyst harvesting proposes, beyond to notice to the survival and
total offspring factors of each population in farm culturing Artemia, the size of cyst that can be produced by each population
already must be check empirically.

FATTY ACID COMPOSITION OF Artemia urmiana IN FIVE SEPARATED STATIONS FROM
THE URMIA LAKE
Ramin Manaffar*, Alireza Asem, Amin Eimanifar and Behrooz Atashbar
Artemia and Aquatic Animals Reserch Institute
Urmia University
Urmia, Iran
raminmanaffar8@yahoo.com
Physico-chemical parameters are very important factors to determine the characteristics of Artemia populations in different
locations. So, widely competitive studies were being done about characterization of Artemia in diverse geographical habitats.
In resent study, percentages of fatty acids of A. urmiana had been analyzed in separated stations from the Urmia Lake. With
regard to results, percentages of fatty acids have variation between studied stations. By the way, Pca analyses shows that there
is no geographical arrangement between disserted sites. This obtained can be led us that ecological parameters are main factors
to conclude Artemia characteristics and cause ecological speciation in this genus.
20

210
REPLACEMENT OF FISHMEAL WITH POULTRY BYPRODUCT MEAL IN DIETS FOR
JUVENILE BLUEGILL Lepomis macrochirus
Karthik Masagounder*, Robert S. Hayward and Jeffre D. Firman
Department of Fisheries & Wildlife Sciences
University of Missouri
Columbia, MO 65211 USA
kmb6b@mizzou.edu
A 60-d trial was conducted to determine the percentage of fish meal that could be replaced by poultry byproduct meal (PBM)
in the diet of juvenile bluegills Lepomis macrochirus (~18 g initial weight) without affecting their growth performance. A
control diet contained 54.6% of fish meal (Table 1). Fish meal from the control diet was progressively replaced with PBM at
20% increments from 0% to 100%, resulting in six diets containing PBM at 0% (PBM0), 10.9% (PBM20), 21.8% (PBM40),
32.7% (PBM60), 43.6% (PBM80), and 54.6% (PBM100) levels. Gross energy levels in the six diets ranged from 18.3 MJ Kg -1
(PBM0) to 19.5 MJ Kg -1 (PBM100). Total amino acid content of the diets ranged from 41 % (PBM0) to 43 % (PBM100). Under
a randomized complete block design, bluegills (n=16/group) were housed individually in plastic test chambers and fed twice
daily at 4% (days 0-30) or 3% (days 30-60) of body weight. Pellets from diet 3 (PBM40) with 40% replacement of fish meal
exhibited low water stability due to an error that occurred in the extrusion program. Consequently, fish that were fed this diet
were excluded from statistical analyses. Data analyses on weight gain (g), Wr (%), hepatosomatic index (HSI), viscerosomatic
index (VSI), visceral fat (%) showed no differences (P>0.05, ANOVA) among dietary groups for any of the metrics (Table 2).
Study results indicate that fish meal can be completely replaced by PBM without affecting growth performance or well-being
of juvenile bluegill reared for 60 d.
Bluegills are aggressive fish and are known to form strong dominance hierarchies when held in groups. This often results in a
few individuals acquiring a high percentage of the feed provided, and the remaining fish receiving much less. Consequences
of dominance hierarchy formation include reduced mean growth and feed efficiency, as well as increased size variation among
bluegills in a tank/chamber. Therefore, the current experiment concerning effects of PBM replacement was conducted with
bluegills that were housed individually. However, we observed that some individually housed bluegills were reluctant to feed
initially, and this factor produced poor overall feed efficiency (~0.5). A similar PBM experiment with group-housed bluegills is
underway to determine whether results based on individually-held bluegills will lead to different conclusions than when groupheld
bluegills are evaluated.
Table 1. Formulations of control diet. Table 2. Final weight (g), weight gain (g), viscerosomatic index (VSI), hepatosomatic
Ingredients g Kg -1
Fish meal 545.5
PBM 0.0
Wheat 246.4
SBM 120.0
Fish oil 74.7
Lecithin 3.0
Vitamin premix 2.5
Dicalcium
2.0
phosphate
Choline-Cl 1.4
Mineral premix 1.0
Vitamin C-PP 0.7
Binder 2.8
Variable
index (HSI), final relative condition factor (Kn), and visceral fat content (%) of juvenile
bluegill fed the the five experimental diets for 60 days. Values are presented
PBM0
(n = 15)
as means±SE.
* n =3 for visceral fat analysis
PBM20
(n = 16)
PBM60
(n = 15)
PBM80
(n = 16)
P-value
PBM100
(ANOVA,
(n = 14)
proc glm)
Final weight (g) 48.8±3.2 44.2±2.0 41.0±1.9 42.7±2.2 43.6±2.2 0.36
Weight gain (g) 30.4±2.9 25.9±1.6 24.8±1.8 26.1±2.2 25.4±2.1 0.49
VSI (%) 12.3±1.1 14.9±1.0 14.0±1.1 14.3±1.5 15.0±1.6 0.33
HSI (%) 0.9±0.1 0.4±0.1 1.1±0.1 0.5±0.1 1.0±0.1 0.33
Wr (%) 109.9±2.5 106.9±2.8 107.8±2.5 108.1±3.3 106±2.0 0.90
Visceral fat (%)* 46.7±2.2 39.5±3.1 44.5±2.8 47.3±1.5 42.4±0.9 0.17

VERIFICATION OF THE POTENCIAL AQUICOLA OF THE SEAWEEED Gracilaria birdae, IN
THE STATE OF CEARÁ-BRAZIL
Toivi Masih-Neto*, Luciana A. A. de Castro and Masayoshi Ogawa
Universidade Federal do Ceará
Departamento de Engenharia de Pesca
Campus do Pici
Caixa Postal: 12.168, CEP: 60.356-000
Fortaleza- Ceará - Brasil
masihneto@bol.com.br
In general the algae are used in several ways, as direct feeding and as for products processed as it is the case of the ficocolóides.
The demand and the readiness of this resource are the main factors that influence in the prices and in the economical viability
of these products. The readiness of some species of algae have been improved through the cultivation, this was introduced in
different places of the continents in sowing way and crop, as the cultivations of Eucheuma in Philippines, Indonesia and Tazânia
and the cultivation of Gracilaria in Chile. Other factors that determine the viability of the cultivations of algae include the
hand of specialized work, hand of organized productive work, equipments, drying, transport, chemical reagents and readiness
of water.
With the objective of verifying the viability and the potential of some areas coastal of the state Ceará, an experimental cultivation
of algae was accomplished, with the species Gracilária birdae, it was observed adaptations in the cultivation structure, the
growth of the algae and the environmental impact, among other aspects in the cultivation.
The cultivation was accomplished parallel in the beaches of Fleixeiras and Guajiru, both in the municipal districts of Trairí-
Ceará-Brazil. Owed the environmental conditions of the place be of relatively busy sea and significant depth, she opted to use
structures type “long-line”, composed by a main rope and secondary strings disposed along the main, tends the extremities of
the fixed structure to the substratum. The experiment was accomplished in partnership with the local residents’ association,
that you/they observe for two years, with collection of data every 15 days, after that period he/she was considered a size and
medium time of crop for that species and cultivation method.
The obtained results were exciting, tends a superior growth that other species of the same gender cultivated at other countries,
the solitary seedlings of the natural bank had medium weight between 50g and 70g and after a cycle of 2 months those reached
medium weight of 750g, with approximate biomass earnings of 1250%, in that period. several aspects in the cultivation methodology
and in the handling of those can be gotten better, still reaching better productivity, however the cultivation was shown
viable, could come to be an excellent alternative of income and job for the fishing communities of the state.
211

212
INITIAL SEEDING WEIGHT AND ALGAE ORIGIN TO MAXIMIZE ALGAE PRODUCTION
Gracilaria sp. IN NORTHEAST BRAZIL
Toivi Masih Neto*, Marcelo Bandecchi B. Miranda and Masayoshi Ogawa
Laboratório de Recurso Aquáticos
Dep.. Eng Pesca
Universidade Federal do Ceara
Campus do Pici s/n, Fortaleza, Ceará, Brazil
masihneto@hotmail.com
Lobster fishery has decreased dramatically in the last few years in Northeast Brazil and aquaculture has proven to be an important
alternative to reduce social effects. The production of macroalgae has shown excellent potential in Ceara State, but productivity
has varied considerably among different attempts and different areas. In order to preserve the natural occurring algae
banks, producers have been advised to start new grow out long-lines with algae form harvested long-lines or with stranded
algae, however many complains have been made that the productivity would be inferior. The objective of the present study was
to investigate the ideal initial seeding weight to maximize production and to evaluate algae growth rate from different origin.
Between May and August 2008 two experiments were conducted in Icapui, Ceara State. In the first experiment, algae portions
of 50 grams were seeded 20 cm apart in 30 meters long-lines. Algae used to start the experiment was collected from five different
sources: (1) from natural algae bank, (2) stranded algae, (3) first time re-seeded from grow out long-lines, (4) second time
re-seeded and (5) third time re-seeded. Samples were taken at 15, 30, 45 and 60 days to evaluate growth rate and productivity.
The second experiment was also conducted in 30 m long-lines and algae were initially seeded at 50, 75 and 100 grams. All
algae used in the experiment came from a nearby natural bank. The results indicate that all re-seeded treatments grew faster than
stranded algae and algae from the natural bank until day 30, however after that the algae from the bank showed extremely fast
growth achieving its peak at 45 days with 405g. At day 30 average algae chunk was 185g for the stranded algae, 191g for algae
from the bank, 225g for the third time re-seeded algae, 274g for the second time re-seeded algae and 329g for the first time
re-seeded algae. At day 60 several algae chunks had detached and dropped from the hope due to overweight, what affected
mainly the fastest growing treatments. Ephinea sp. fouling was also very consistent after 45 days. The second experiment
showed that there is a trade-off between the initial seeding weight and time of harvest. Algae that started with 50g achieved
maximum productivity at 45 days (368g) and declined after that. Although algae that started with 100g showed heavier average
than the others at day 30 (347g) and day 45 (425g) the weight difference and harvest time was not enough to compensate
initial seeding size. Seeding with half the amount (50g) is enough to seed twice as much long-lines, therefore profitability
should be higher as the final weight and harvest time do not differ considerably. We also concluded that seeding with algae from
previously harvest long-lines is viable up to the third re-seeding, but productivity declines as the same algae are used several
times. Algae from natural banks or stranded take longer to launch their growth probably due to the time needed to produce new
extremities, but once this is achieved the growth is accelerated.

AQUAPLANT: A WEB-BASED TOOL FOR AQUATIC PLANT MANAGEMENT
Michael P. Masser and Satoshi Kamata
111B Nagle Hall
]2258 Tamu
College Station, Texas 77843-2258 USA
m-masser@tamu.edu
Aquatic plants are a problematic issue for most aquaculturists. To assist aquaculturists and other private impoundment managers
a web-base identification and management tool (i.e. AQUAPLANT) was developed for the Texas Agricultural Extension
Service (i.e. Texas AgriLife Extension) in 2000. AQUAPLANT went through a major revision in 2005 and continues to be
updated semi-annually.
The AQUAPLANT site consists of 72 species/families of aquatic plants common to the Southern states and most of the U.S.
For each specie or group the site assists in Identification through photographs, drawings, and simple descriptions. Once identification
is made, Management Options are provided for possible Mechanical, Biological, or Chemical control. Links are
provided to MSDS herbicide labels for all the major aquatically registered (i.e. EPA) herbicides.
AQUAPLANT has proven to be a heavily used resource with over 152,000 unique visitors in 2007 and 130,000 in 2008 (i.e.
Jan-July), downloading 664,440 pages in 2007 and over 650,000 so far in 2008. A user satisfaction on-line survey was conducted
in 2007 (June – December). The survey reviled that the site was used mostly by private impoundment owners to identify
a problem plant, although in most cases no management was to be applied. However, satisfaction was good as users were
“Highly Likely” to recommend the site to others. AQUAPLANT has proven to be a useful tool for aquatic plant identification
and management.
213

214
SUSCEPTIBILITY OF SPOTTED SEATROUT Cynoscion nebulosus AND RED SNAPPER
Lutjanus campechanus TO Amyloodinium ocellatum INFECTIONS
Ignacio Masson*, Jeffrey M. Lotz and Reginald B. Blaylock
Gulf Coast Research Laboratory
The University of Southern Mississippi
Ocean Springs, MS 39564 USA
ignacio.masson@usm.edu
Amyloodiniosis, a fish disease caused by the parasitic dinoflagellate Amyloodinium ocellatum, is one of the most devastating
diseases in warm water marine aquaculture. The life cycle of A. ocellatum consists of a parasitic trophont, a free-living, dividing
tomont and a free-swimming infective dinospore. Trophonts attach to the gills and skin, diminish the gills’ functional surface
area, damage the epithelium, and cause fish death. Different fish species have different susceptibilities to infection reflected in
dinospore infection rates (the proportion of dinospores that attach to the host in 24h and become trophonts), dinospore lethal
doses (the number of dinospores that results in host death), trophont lethal loads (the number of trophonts on a host after a
challenge with a lethal dinospore dose) and trophont detachment rates (the proportion of trophonts that detach from a host per
time unit after a single challenge with dinospores). In this study we compared the susceptibility of juvenile spotted seatrout
(16.36±0.82 g, mean±SE) and juvenile red snapper (17.15±2.04), two fish species with potential for culture in the Gulf of
Mexico, to A. ocellatum infections. The mean infection rates (24-h individual exposures in 3-l aquaria) for spotted seatrout and
red snapper were not significantly different (0.3552±0.0137 and 0.3414±0.0147 dinospores/d respectively, mean±SE; p>0.05).
However, there was a significant difference (p

EFFICIENT OXYGENATION FOR SUSTAINABLE AQUACULTURE
Dennis L. Mast, PhD
Clean Water Scientific, Inc
1121 Situs Court, Suite 120
Raleigh, NC 27606 USA
dmast@cleanwaterscientific.com
The technology made for Efficient Oxygenation is a device known as the “Add-o-mizer” and is specifically made for dissolving
gas into liquid. The Add-o-mizer is comprised of a pressure vessel or column for receiving a gas-entrained liquid via an inlet
and for injecting the gas-entrained liquid via a riser into a headspace of the vessel. A flow director is disposed in an upper portion
of the vessel or column to form a swirling flow path extending into a liquid pool in a lower portion of the vessel or column.
An outlet is provided to direct the liquid away from the vessel or column into a non-pressurized body of water
As the processed effluent is sufficiently supersaturated with oxygen, excess dissolved gas will flash to vapor or micron sized
bubbles upon return to the non-pressurized tank or basin. By controlling the level of super saturation, the size of these “microbubbles”
can be controlled to less than 5 microns in diameter - 0.005 millimeters! Slow rise rates of 0.2 to 0.8 cm/sec combined
with a surface-to-volume ration more than 1000 times higher than that for fine bubble diffusers. This makes microbubbles a
very efficient mechanism of oxygenation with minimal operating and maintenance costs.
The system under pressure can achieve dissolved oxygen (DO) levels far greater than possible with a mechanical aerator in an
open basin. The Add-o-mizer aerates by dissolving the feed gas into the water under pressure. This often makes aerating with
air, rather than oxygen, very cost effective.
Other types of aeration systems operate at relatively low efficiencies. As a result, air must be supplied in great excess in order
to ensure adequate dissolution of oxygen. With a (generous) oxygen transfer efficiency of 15%, a blower system must pump
374 cubic feet of air into water to dissolve 1 pound of oxygen. In order to dissolve the same 1 pound of oxygen into water, the
technology only needs to inject 11 cubic feet of air.
Benefits of this technology are efficient oxygenation, less feed requirements, shorter maturation cycles and healthier aquaculture
by using ambient air instead of being required to use liquid oxygen. If necessary, the tank/pond water can be sanitized
following each cycle/harvest without the need to replace all of the water by using the Add-o-mizer along with ozone as the gas
being entrained into the liquid.
21

21
EFFICIENT OXYGENATION FOR TIDAL AQUACULTURE
Dennis L. Mast, PhD
Clean Water Scientific, Inc
1121 Situs Court, Suite 120
Raleigh, NC 27606 USA
dmast@cleanwaterscientific.com
The technology made for Efficient Oxygenation is a device known as the “Add-o-mizer” and is specifically made for dissolving
gas into liquid. The Add-o-mizer is comprised of a pressure vessel or column for receiving a gas-entrained liquid via an inlet
and for injecting the gas-entrained liquid via a riser into a headspace of the vessel. A flow director is disposed in an upper portion
of the vessel or column to form a swirling flow path extending into a liquid pool in a lower portion of the vessel or column.
An outlet is provided to direct the liquid away from the vessel or column into a non-pressurized body of water.
The system and method for dispersing oxygen into a submerged fish cage in tidal water is provided where the tidal movement
of the water is taken into account and oxygen, or air containing oxygen, is strategically dispersed in the fish cage such that the
tidal movement of water through the fish cage tends to efficiently distribute the oxygen for use by fish within the cage.
Disposed in opposite ends of the cage is a pair of oxygen diffusers where each oxygen diffuser extends across an opposed end
of the cage. In this embodiment, each oxygen diffuser is disposed adjacent to the bottom of the cage but along an extreme
boundary of the cage. Each oxygen diffuser is an elongated conduit or pipe with openings formed in the outer wall thereof. As
will be discussed subsequently herein, the function of the oxygen diffusers is to disperse an air-water mixture into the cage for
supplying oxygen to the fish contained within the cage.
The Add-o-mizer is operated as a device where water is pumped into it under pressure and dissolves gas into liquid. The device
includes a pressure vessel having an inlet disposed on the pressure vessel for receiving a gas-liquid mixture. A riser is disposed
in the pressure vessel and connected to the inlet. The riser extends into a head space of the pressure vessel. The riser is adapted
to receive the gas-liquid mixture from the inlet and inject the mixture into the head space. An opening is disposed in an upper
end of the riser below an interior surface of the pressure vessel. Disposed in an upper portion of the pressure vessel is a flow
director that forms a swirling flow path. An outlet is disposed on the pressure vessel for directing the liquid from the pressure
vessel.
Benefits of this technology are efficient oxygenation, less feed requirements, shorter maturation cycles and healthier aquaculture
by using ambient air instead of being required to use liquid oxygen.

PRODUCTION OF Artemia CYSTS AND FLAKES USING IMPROVED TECHNIQUES IN
ROCK SALT SOLUTION
San Ei Ei Maung*, Dr. Hnin Hnin Htun and Dr.Mar Too Nyi Bu
Aquaculture Biotechnology Research Center
Department of Biotechnology
Mandalay Technological University
Myanmar
The high quality (disinfected and good nutrient level) Artemia cysts could be firstly produced in our country using rock salt
solution without adding other chemicals. Initially, the seven Artemia cysts brands were hatched to know the best and suitable
brand. Secondly, many ways of disinfection and decapsulation procedures were tested depending on hatching efficiency.
Thirdly, Artemia were cultured depending on the effects of variable feeds (energy and protein source). For this, there were
three experiments, the first one was the use of rice bran suspension, the second was the use of yeast + vitamins suspension,
and the third was the use of rice bran + yeast + vitamins suspension. After one month, it was found that the third was the fastest
metamorphosis and the highest cysts production. Fourthly, the disease challenge tests were made using bioencapsulation
method (boosting). For this, there were four experiments, the first one was natural (control), the second was the use of neem
leaf decoction, the third was the use of probiotics, the forth was the use of antibiotics. After one month, the lowest bacterial
count and highest survival rate were showed by the second and third. Finally, by using the best data from the above experiments,
high quality Artemia cysts were produced. The hatching efficiency of this produced cysts were tested two times per
month and showed the best data. As the by-products from cysts production, Artemia flakes (protein 53%) were produced for
post larvae. So, this Artemia cysts and flakes production technology will be suitable for the places where cannot find naturally
Artemia cysts and cannot use natural seawater to produce marine aquatic feed products (Artemia cysts and flakes) and will effective
not only aqua-farmers but also salt-farmers by combination with solar salt and Artemia cysts production technology to
get extra-income.
21

21
CONTRIBUTION POTENTIAL FROM WINTER BURROWING BROODSTOCK IN
LOUISIANA CRAWFISH PONDS
W. Ray McClain
Rice Research Station
Louisiana State University Agricultural Center
1373 Caffey Road
Rayne, Louisiana 70578 USA
rmcclain@agcenter.lsu.edu
Because crawfish aquaculture in the South depends on natural recruitment to populate ponds, and because harvest and recruitment
are continual, many producers are apprehensive about harvesting mature females early in the season for fear of adversely
affecting subsequent recruitment and thus, yields. Some farmers will delay harvest or return captured mature females back to
the pond during early harvests – typically a time of highest marketing prices – to ensure the greatest possible production. Aside
from the low efficiency of trap harvests, information on winter reproduction in Procambarus clarkii is sufficiently lacking to
justify this practice. Hence, this study was initiated to ascertain the typical spawning dates for winter burrowing crawfish.
P. clarkii dominate commercial crawfish ponds in Louisiana and have evolved to reproduce in burrows rather than in open
waters. For this reason, freshly constructed burrows along the levees of experimental crawfish ponds were flagged during the
winter months over a 2-year period, and then excavated at the end of each month. The unearthed females were placed in artificial
burrows in a lab that closely simulated conditions of the natural burrow, and were examined monthly for spawning activity.
Results are presented in the table below.
Spawning dates were positively correlated to month of burrowing, and it is assumed that maturation and spawning were little
affected by the move to the lab. Although no fresh burrows were observed during December in year 2, it appears that crawfish
that burrow later than December will not likely contribute offspring to the harvestable population within the current season
that typically ends by mid June in Louisiana. Hatchlings are not released until at least 5 weeks after spawning and because
January burrowers did not spawn until mid March or later in this study, it is unlikely such hatchlings could reach market size
in large numbers before ponds are drained. Therefore, it may be futile for Louisiana producers to protect mature females from
the harvest after about December, if at all, in commercial crawfish ponds.
Table. Mean spawning period and young hatched from crawfish burrowing during the winter months in
Louisiana crawfish aquaculture ponds.
Avg.
Ind.
Spawning
Range
Year 1 Year 2
Avg.
Month 1 of
No.
Young
per
Avg.
Ind.
Spawning
Range
Avg.
Month 1 of
No.
Young
per
Overall
Means for
Month of
Spawning
Month of
Burrowing Wt. (Months) 1
Spawning gram Wt. (Months) 1
Spawning gram 2
Dec 19.4 2 - 3 2.5 18.8 - - - - mid Feb
Jan 15.9 3 - 5 4.0 16.0 20.3 3 - 4 3.4 11.5 late Mar
Feb 25.6 4 - 5 4.2 10.5 23.8 4 - 6 4.7 9.6 mid Apr
Mar 25.6 4 - 8 5.7 15.5 20.8 5 - 6 5.2 - mid May
1
Months are indicated by numerical representation, beginning with 1 for January.
2
Young died before release from all females of the March group in Year 2.

VARIATION OF EGG SIZE OF DOMESTIC WHITE BASS Morone chrysops DUE TO
DIETARY DIFFERENCES
Matthew McEntire* and S. Adam Fuller
USDA / ARS – H.K. Dupree Stuttgart National Aquaculture Research Center
P.O. Box 1050
2955 Hwy 130 East
Stuttgart, Arkansas 72160-1050 USA
Matthew.mcentire@ars.usda.gov
Tagged domestic three year old white bass brood fish were stocked in 5500 L tanks at the Harry K. Dupree Stuttgart National
Aquaculture Research Center with flow through well water mimicking a normal thermal and diurnal pattern in the South Eastern
USA. There were two groups of fish fed different diets during most of vitellogenesis. The control group was fed a 45%
protein 16% fat diet containing primarily fish meal and the other was fed a 45% protein, 15% fat diet containing fish meal and
squid meal. Fish were of the same age and genetic stock but held at different facilities prior to stocking. The mean (+ S.E.) fish
weights were 714+121g for the control group and 558+105g for group fed the diet containing squid meal and were statistically
different (p< 0.0001). In April fish were injected with a slow release gonadotropin-releasing hormone (GnRH) pellet to induce
oocyte maturation and ovulation.
Eggs were collected from 54 white bass females and preserved in 70 % isopropyl alcohol. All the egg samples were treated
the same and the amount of shrinkage in egg size was expected to be uniform and constant across groups. The eggs were later
photographed and mean relative egg size was measured to the nearest mm (n=30). The mean egg diameter was 0.85+0.05mm
for the control and 0.86+0.06mm for the diet containing squid meal. There was no statistical difference in egg sizes between
the groups (p

220
SUCCESSFUL APPLICATION OF A SCIENCE-BASED APPROACH TO SETTING
REGULATORY ENVIRONMENTAL STANDARDS FOR COASTAL FINFISH
AQUACULTURE, BRITSH COLUMBIA, CANADA
Eric R. McGreer* and Bernie Taekema
British Columbia Ministry of Environment
Environmental Protection Division
Regional Operations, Vancouver Island
2080A Labieux Road
Nanaimo, BC, Canada V9T 6J9
eric.mcgreer@gov.bc.ca
In September 2002, a new performance-based, provincial waste regulation was introduced in British Columbia, the Finfish
Aquaculture Waste Control Regulation (FAWCR). The FAWCR was developed to provide protection for the marine environment,
and support lifting of an existing moratorium on new coastal fish farm licenses. Development of the chemical and
biological standards used in the regulation was based on extensive pre-monitoring and utilization of existing scientific
knowledge from the Pacific Northwest and globally. An overview of the results from seven years of environmental monitoring
will be presented together with a discussion of key factors which have contributed to the success of the program. The FAWCR
is currently being amended to incorporate new scientific information, and expand and streamline its role in environmental
protection while accommodating increased production efficiencies by the industry. The importance of the amendments will be
discussed in relation to their implications for regulatory science.
The primary chemical standard used in the FAWCR is the concentration of free sulphides in sediment pore water. Specific
levels not to be exceeded are provided for the fish production cycle including peak production (6,000 uM) and re-stocking
(1,300 uM). Exceedance prompts biological sampling of benthic communities to confirm impact. Generally, compliance with
the FAWCR by the industry since 2002 has been good which has required the industry to actively manage its operations to
accommodate increases in production while keeping within the regulatory limits. Changes in benthic community structure at
farms which have demonstrated impacts over several years have been documented, and the steps to development of indicator
species will be highlighted.
Research in support of regulatory amendments has been completed in the areas of development of protocols and criteria for
video monitoring of hard seabed substrates, adoption of an “ecological threshold” approach to biological monitoring, updating
protocols for use by government agencies and industry, increasing regulatory effectiveness to balance environmental protection
and increased production efficiencies by industry, and certification of data/information providers.
Emerging issues which require regulatory research for possible inclusion in future amended versions of FAWCR have been
identified as far field effects, application of an ecosystem-based approach, and several forms of cumulative effects.

BAITFISH AQUACULTURE IN THE MID-ATLANTIC: PRODUCTION OF MUMMICHOG
Fundulus heteroclitus IN FRESH WATER PONDS
Dennis McIntosh
Delaware State University
1200 N. DuPont Hwy.
Dover, DE 19901 USA
dmcintosh@desu.edu
This project was initiated in 2005 with the goal of determining the production potential of mummichogs (Fundulus heteroclitus)
in inland, freshwater ponds. Mummichogs, also know as mud minnows or bull minnows, are a local estuarine species that
are popular as live bait for flounder, speckled trout, and red drum. Many of the other common bait species found in the bait
shops locally are freshwater species, usually either golden shiners or fathead minnows, and are not suitable for use in the salt
water. Historically, supplies of freshwater baitfish are farmed and trucked in from other states, while mummichogs are trapped
locally. Unfortunately, as demand for mummichogs and other saltwater bait species increase, new supplies are needed.
Culture techniques for golden shiners and fathead minnows are well established. Conversely, much research is needed before
saltwater baitfish aquaculture reaches viable industry status. We have identified the following objectives to be researched in
order to begin to develop practical culture technologies for mummichog production in the mid-Atlantic.
1. Compare egg-transfer to fry-transfer production systems for mummichogs (Year 1).
2. Determine optimum mummichog stocking density (Year 2).
3. Evaluate the effects of dietary protein on mummichog production (Year 3).
4. Compare batch, versus continuous harvest methods (Year 4).
Over the four years of this project, survival was extremely variable from year to year, making interpretation difficult. Regardless,
a number of interesting trends are evident. Details of each year’s research and the corresponding results will be presented.
In addition, future research goals and objectives will be discussed.
221

222
DETERMINING THE EFFECT OF THREE WATER CONDITIONERS ON HOLDING
SUCCESS OF MUMMICHOG Fundulus heteroclitus IN SIMULATED COMMERCIAL
HOLDING TANKS
Dennis McIntosh*, Adrienne George
Delaware State University
1200 N. DuPont Hwy.
Dover, DE 19901 USA
dmcintosh@desu.edu
Each year U.S. farmers produce $38 million worth of live bait for sport fishing. The two most common species produced,
golden shiners (Notemigonus crysoleucas) and fathead minnows (Pimephales promelas), are used as a bait when fishing in
freshwater. In the Mid-Atlantic however, there is also a strong demand for saltwater bait that can be used when fishing in estuarine
and marine waters. Mummichog (Fundulus heteroclitus) are considered an important local baitfish species because it can
be used as saltwater bait, where they are commonly used to catch flounder, speckled trout, and red drum. Currently supplies of
this species are limited to wild harvest, although there is work underway to develop production techniques for mummichogs.
After being harvested, mummichogs are held at high densities in retail holding tanks at bait shops for sale to recreational anglers.
Water quality deterioration in the retail holding tanks can result in a significant level of mortality and/or reduced bait
quality. To help mitigate this problem, a number of water conditioners are available that claim to provide stress relief and
thereby improve holding success.
The effects of three water conditioners on water quality and survival of mummichogs held under simulated retail conditions
was studied over 23 days at the Delaware State University Aquaculture Research and Demonstration Facility in Dover, DE.
Sixteen 163-L fiberglass tanks were each filled with 96.4 liters of water and 1,700 grams of fish (~ 400 fish/tank). One of four
treatments was then randomly assigned to each of the 16 tanks (4 replicates/treatment).
Results from this study indicate that using either of the salt treatments (CaCl or NaCl) was a better choice then either the control
or the commercial water conditioner, Better Bait, to improve the holding success of mummichogs held under simulated retail
conditions.
Water conditioner treatments applied to simulated retail bait holding tanks
over 23 days.
Treatments Dose Effective Concentration
Control (C)
(g/tank)
(g/L)
Sodium Chloride (NaCl) 280 2.96
Calcium Chloride (CaCl) 280 2.96
Better Bait * 7 0.07
*Sure-Life Laboratories Corporation, Seguin, TX

Aeromonas hydrophila SEPTICEMIA IN IMPORTED LIONHEAD GOLDFISH IN IRAN
S.Mehdizadeh Mood and H.Ebrahimzadeh Mousavi
Department of Aquatic Animal Health
Faculty of Veterinary Medicine
Tehran University
Bacterial disease is the most common infectious problem of ornamental fishes, and most bacterial infections are caused by
gram-negative organisms. Extrinsic stressors, including shipping, crowding, poor water quality, and inadequate nutrition, may
predispose an ornamental fish to bacterial disease.
High mortality rate in one of the importing ornamental fish farms in Tehran occured in June 2008. To recognize the reason 50
samples of fishes with clinical sign including anorexia ,Exophthalmia , hyperemia and hemorrhage in skin and fins associated
with fin rot and erosion in some cases were taken to the lab.
In first stage samples were examined for parasite infection and in wet smear 1-3 Gyrodactylus sp. have been observed .but
according to the clinical sign and the high mortality rate , such a light infestation couldn’t be the reason. In necropsy , fluid in
abdominal cavity associated with hyperemia in internal organs and necrosis have been observed that show a bacterial septicemia.
In bacterial culture the bacteria which isolated from the samples was Aeromonas and biochemical results show that the
species is Hydrophila.
Reduction in number of fish per litre ,water exchange and Oxytetracycline (50 to 75 mg/kg of fish for 10 days) which has been
the drug of choice for treating motile aeromonad septicemias in fishes were recommended that lead to considerable reduction
of mortality rate.
223

224
HISTORY AND PRESENT STATUS OF PADDLEFISH AQUACULTURE IN RUSSIA
Evgeniy Melchenkov, Vladimir Vinogradov, Marina Mihailova and Boris Gomelsky
All-Russian Research Institute of Freshwater Fisheries
Rybnoe, Moscow Province
141821 Russia
vniprh@mail.ru
Russia has a long history of sturgeon culture. However, all sturgeons which inhabit local waters are bentophagous or predators.
Therefore, the interest towards American paddlefish (Polyodon spathula), the only representative of order Acipenciformes
that consume zooplankton, was enormous. For the first time paddlefish were delivered from the United States in 1974 as a
result of scientific exchange between USSR Government and US Fish and Wildlife Service.
For successful introduction of this species to Russian aquaculture, intensive long-term studies were performed:
• investigated functional structure of filtration apparatus and paddlefish feeding selectivity in ponds;
• investigated gametogenesis and maturation cycles;
• investigated embryonic development and influence of different factors on survival of embryos;
• developed methods for rearing of broodstock;
• developed methods of artificial breeding;
• developed methods of fingerling production;
• developed methods of rearing of paddlefish up to the market size;
• evaluated the nutritional value of paddlefish.
Based on results of these studies the technologies for artificial reproduction and rearing of paddlefish were developed. Paddlefish
are raised in fish farm ponds or stocked into reservoirs and lakes for ranching. Currently paddlefish broodstock are kept at
two locations in the south of Russia (Astrakhan and Rostov regions). Under local environmental conditions paddlefish males
reach sexual maturity at the age of 6-7 years while paddlefish females become mature at the age of 10 years. Sturgeon pituitary
extracts are injected for induction of final oocyte maturation and ovulation in females and spermiation in males. The same
incubators used for sturgeons (Yushenko, Osert etc.) are also used for incubation of paddlefish embryos. Paddlefish larvae are
raised in ponds or tanks using zooplankton. When raised in ponds in polyculture with sturgeon or common carp, paddlefish
reach market size (1.5-2 kg) in two years. Currently in Russia paddlefish research and development have advanced to commercial
production at fish farms. Introduction of American paddlefish to Russia has successfully developed a new niche market.
American paddlefish produced in Russia have been delivered to many other countries of former Soviet Union as well as to
China and Cuba. Technologies on artificial reproduction and culture of paddlefish developed in Russia have been used in many
countries for development of their own broodstock.

PRELIMINARY NUTRITIONAL INVESTIGATIONS OF THE SPOTTED SEATROUT
Cynoscion nebulosus
M. Shane Merryman*, Joe M. Fox, William H. Neill and Robert Vega
Harte Research Institute for Gulf of Mexico Studies
6300 Ocean Drive, Unit 5869
Corpus Christi, TX 78412-5869 USA
michael.merryman@tamucc.edu
In terms of recreational fishing, the spotted seatrout
(Cynoscion nebulosus) is one of the leading sports
fish in the U.S. With this popularity, a high potential
for overexploitation exists which could ultimately
diminish wild stocks. In Texas, state fisheries managers
have initiated a stock enhancement program along
the natural range of this species to supplement wild
stocks. This implies possible future need for aquaculture
of this species. With this in mind, two studies
were undertaken to address the basic nutritional needs
of spotted seatrout: 1) to determine feed rate for maximum
weight gain and maintenance and 2) to evaluate
the effect of involuntary exercise on growth. Six feed
rates (1-6% of total tank biomass/d, bwd) were evaluated
in triplicate tanks (n =18) for a period of 5 wk. A
total of 15 fish (mean initial weight = 1.4 ± 0.1g) were
stocked into each tank. Results showed significantly
higher weight gain (P

22
EVALUATION OF DIETS CONTAINING SOYBEAN MEAL, WITH AND WITHOUT AMINO
ACID SUPPLEMENTATION, AS TOTAL REPLACEMENT FOR FISH MEAL FOR JUVENILE
NILE TILAPIA, Oreochromis niloticus
Linda S. Metts*, Kenneth R. Thompson and Carl D. Webster
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
linda.metts@kysu.edu
Global production of tilapia has increased dramatically over the past few years due to new technologies and research that have
allowed for expansion of aquaculture production. World-wide tilapia production is expected to reach 4 million tonnes in the
near future. However, with the use of fish meal in aquaculture diets being questioned by some as either environmentally and/or
financially unsound, the need for partial or total replacement of fish meal with plant and animal alternatives is essential for
the future growth of the aquaculture industry. Therefore, the objective of the study was to evaluate the use of a less-expensive
protein source (soybean meal), with and without amino acid supplementation, as total replacement of fish meal (FM) in diets
for Nile tilapia (Oreochromis niloticus).
An 8-week feeding trial will be conducted on small (100-200 mg) Nile tilapia obtained from a commercial supplier. Nile tilapia
will be fed nine experimental diets, with eight experimental diets formulated to contain either all soybean meal (with and
without methionine and/or lysine supplementation), or a blend of soybean meal and either poultry by-product meal or distillers
dried grains with solubles. The ninth diet will be formulated to be similar to a high-quality commercial tilapia diet which will
contain 20% menhaden fish meal and 40% protein. Weight gain, survival, and body composition will be measured at the conclusion
of each 8-week feeding trial. Since the future of long-term availability of FM is uncertain, and since FM is one of the
most expensive ingredients in an aquaculture diet, it is important to the aquaculture industry that production diets be formulated
using alternative ingredients that can replace or reduce the percentages of FM without adversely affecting growth and health of
fish. The results of this study may give new insights on possible alternative protein sources that can replace FM in Nile tilapia
diets as well as other omnivorous and/or carnivorous species, which may allow further expansion of the aquaculture industry.
COMPARISON OF MIST AND MODIFIED-MIST PROCEDURES FOR QUICK EGG
REMOVAL FROM OVULATED PADDLEFISH
Steven D. Mims*, Richard J. Onders and Boris Gomelsky
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
steven.mims@kysu.edu
A procedure known as Minimally Invasive Surgical Technique (MIST) has been successfully evaluated and practiced for quick
egg removal from ovulated paddlefish. Though the technique requires little out-of water handling time for the fish and results
in high egg yields and broodstock survival, a modified MIST will be presented to show an alternative procedure that is as safe
for the brood stock but even quicker in egg removal. Embedded video will be used to illustrate and to compare between the
two MIST procedures.

HYDROGEN PEROXIDE TREATMENTS FOR CHANNEL CATFISH EGGS INFECTED
WITH WATER MOLDS
Andrew Mitchell*, David Straus, Andy Radomski and Ray Carter
Harry K. Dupree Stuttgart National Aquaculture Research Center
United States Department of Agriculture
Agriculture Research Service
P. O. Box 1050
Stuttgart, Arkansas 72160 USA
drew.mitchell@ars.usda.gov
Fungi, or water molds Saprolegnia spp., on channel catfish Ictalurus punctatus eggs can lower fry production. This requires
the producer to spawn more catfish or face fingerling shortages. Few treatments have been tested against channel catfish eggs
infested with an identified fungus. Hydrogen peroxide treatments were evaluated to determine their effect on egg survival and
their efficacy against naturally occurring Saprolegnia spp. infestations.
Three samples (12.8 g average) taken from each egg mass (less than 24-h old) were weighed and the eggs counted to determine
the number of eggs per gram (overall average of 28.8 eggs/g). Egg masses were separated into approximately 90 g portions and
placed in hatching troughs with nine separate compartments; each compartment had its own water inlet, drain and paddle. The
compartments contained 35 L of water with an inflow of about 1.25 L/min (28 min water exchange rate). Water temperatures
(23.2-24.0°C) were monitored continually, dissolved oxygen (4.5 to 6.4 mg/L) and water flows were taken daily (adjusted if
needed), and total alkalinity (215 mg/L) and total hardness (110 mg/L) were taken at the beginning of each study. Hydrogen
peroxide was applied once daily as a flow through treatment at ten concentrations (0, 15.6, 31.3, 62.5, 250, 500, 1,000, 2,000,
and 4,000 mg/L) in three studies until the eggs were clearly in the eyed-egg stage (5 or 6 treatments). In the last two studies,
identical treatments (0 to 500 mg/L) were applied for 6 d in each study. Observations for fungal development were made
daily until hatching began. The maximum measure of fungal growth (nearest 0.5 cm) was recorded from the egg mass in each
compartment. This value was used as the fungal coverage measurement. Hatched fry, at the termination of each test run, were
counted and percent survival determination (counted fry/number of potential fry X 100) for each treatment was made.
Concentrations above 500 mg/L drastically lowered egg survival and were dropped after the first study. Concentrations of 125,
250 and 500 mg/L were significantly different from the controls and all treatments significantly reduced fungal coverage when
compared to the controls but were not different from each other. The 250 mg/L treatment had the highest average survival
(64.4% compared to 24.7% for the controls), lowest incidence of fungi (27.3% compared to 100% for the control) and the lowest
average fungal coverage (0.95 cm compared to 7.09 cm for the controls).
22

22
SOME BIOLOGICAL CHARACTERISTICS OF CRAYFISH, Astacus leptodactylus (Decapoda;
Astacidae) IN ARAS DAM (2008)
Ali Mohsenpour Azari*, Alireza Asem, Feridun Mohebi and Saber Shiri
Iranian Artemia Research Center (I.A.R.C)
Urmai, Iran
Ali_mohsenpour@ifro.ir
Length- Weight structure and sex ratio of Astacus leptodactylus in Aras Dam have been studied during three seasons (spring,
summer and fall) in 2008. This survey has shown that %52.11 of total catch from Aras Dam was larger than the standard level
of length (=100 mm) while it was %83.2 in 1997-98. As well as %27.11 of total catch was heavier than the standard level of
weight (50 gr.). The total sex ratio (Male: Female) was 1: 0.70. The mean of length and weight were 106.73 ± 20.97 mm and
40.22 ± 30.77 gr. for males and 107.22 ± 14.9 mm and 31.88 ± 12.77 gr. for females respectively. High among of Standard
Deviation, specially about males sample and also the decreased the total the total percentage of standard levels of length and
weight suggest tat the population of Astacus leptodactylus doesn’t have normal and standard conditions for harvesting and
export activities.

NUCLEOTIDE SEQUENCE VARIATIONS OF THE MAJOR STRUCTURAL PROTEINS
(VP15, VP19, VP26 AND VP28) OF WHITE SPOT SYNDROME VIRUS (WSSV), A PATHOGEN
OF CULTURED LITOPENAEUS VANNAMEI IN MEXICO
Zinnia J. Molina Garza, Lucio Galaviz Silva and J. Reyes González Galaviz
Universidad Autónoma de Nuevo León]
Facultad de Ciencias Biológicas
molinazinnia@hotmail.com
Since 1992, white spot syndrome virus (WSSV) has caused severe damage to the global shrimp farming industry and is the
most devastating shrimp viral pathogen currently known. White spot syndrome virus (WSSV) was first reported in farmed
Litopenaeus vannamei stocks in Sinaloa and Sonora, Mexico during 1999 and continues to cause severe shrimp losses.
White spot syndrome virus particles were purified from L. vannamei collected during epizootics from shrimp farms located in
Sinaloa, Nayarit and Sonora, Mexico.
Open reading frame fragments that encode the described proteins were amplified from the purified WSSV Mexican isolate (Fig.
1). The amplified PCR products fitted in the expected positions and corresponded to vp28 (516 bp), vp19 (363 bp), vp26 (569
bp) and vp15 (311 bp).
WSSV genes encoding nucleocapsid (VP26 and VP15) and envelope proteins (VP19 and VP28) of a Mexican isolate were
cloned in the pMosBlue vector. The nucleotide sequences of these genes were compared with WSSV isolates in GenBank.
VP15 is highly conserved, and VP26 showed 99% homology to a Chinese isolate. The VP28 fragment demonstrated 100%
homology to the majority of the isolates analysed (UniProt accession no. Q91CB7), differing from two Indian WSSV and one
Chinese WSSV isolates by two non-conserved and one conserved replacements, respectively. Because of their highly conserved
nature, these three structural proteins are good candidates for the development of antibody-based WSSV diagnostic tools
or for the production of recombinant protein vaccines to stimulate the quasi-immune response of shrimp. In contrast, VP19 of
the Mexican isolate was distinguishable from almost all isolates tested, including an American strain of WSSV (US98/South
Carolina, GenBank accession no. AAP14086). Although homology was found with isolates from Taiwan (GenBank accession
no. AAL89341) and India.
Figure 1 Agarose-gel electrophoresis
showing PCR products of four genomic
regions that encode WSSV nucleocapsid
and envelope proteins. Lane M, DNA
marker, lane 1, vp15 (311 bp); lane 2,
vp19 (363 bp); lanes 3 and 4, vp26; lane
5, vp28.
22

230
EVALUATION OF SOY LECITHIN IN PLANT-BASED DIETS FOR Litopenaeus vannamei
CULTURED IN OUTDOOR TANKS
Timothy C. Morris*, Tzachi M. Samocha, D. Allen Davis, Josh S. Wilkenfeld and Kelsey Holmes
AgriLife Research Mariculture Laboratory
4301 Waldron Rd.
Corpus Christi, TX 78418 USA
tilapiamorris@juno.com
Plant-based feeds present a more environmentally-sustainable alternative to fishmeal-based diets, but require further nutritional
optimization prior to commercial acceptance. Phospholipids (PL) play an important role in maintaining cellular function and
structure. They also enhance cholesterol transport, increase lipid retention, and may facilitate the pelleting process by acting as
a lubricant. Soy lecithin, a primary source for PL, has the potential to improve the lipid component of plant-based feeds. This
study evaluated the response of shrimp to increasing levels of dietary lecithin (0 to 2%) in conjunction with decreasing levels
of fish oils designed to minimize shifts in lipid content of the diets.
The 10-week trial was conducted with juvenile (1.37 ± 0.05 g) Pacific white shrimp, Litopenaeus vannamei, previously conditioned
to a commercial diet (35% crude protein, Rangen, Buhl, Idaho). Shrimp were stocked in a recirculating system
consisting of thirty-six 800-L HDPE tanks (0.65 m 3 water volume) at a density of 26 shrimp/tank (40/m 3 or 31/m 2 ). Water
was recirculated at 1.9 L/min/tank. Naturally-induced primary production was present, aeration was provided by two airstones
(7-10 Lpm/stone), and no external biofiltration was provided. Five plant-based diets were formulated to contain 35%
protein, 6% lipid, and five lecithin levels: 0, 0.25, 0.50, 1.00, and 2.00%. The conditioning feed also served as a reference diet.
Rations were determined based on an assumed growth rate of 1.5 g/wk, an FCR of 1.4, and 100% survival. Feed was offered
three times daily. The study was terminated when shrimp averaged more than 20 g. A one-way ANOVA showed significant
differences between treatments; with average weight, growth, yield, and FCR values ranging from 21.5-22.7 g, 1.9-2.0 g/wk,
0.84-0.90 kg/m 3 , and 1.3-1.4, respectively. Survival was not significantly different between treatments. Results of the present
study indicate a general increase in performance up to 1% inclusion of lecithin.

RESPONSE OF PACIFIC WHITE SHRIMP, Litopenaeus vannamei, TO VARYING RATIOS
OF SOYBEAN OIL AND FISH OIL IN PLANT-BASED DIETS OFFERED TO SHRIMP
CULTURED IN OUTDOOR TANKS
Timothy C. Morris*, Tzachi M. Samocha, D. Allen Davis, Josh Wilkenfeld and Kelsey Holmes
Texas AgriLife Research Mariculture Laboratory
4301 Waldron Rd.
Corpus Christi, TX 78418 USA
tilapiamorris@juno.com
Plant-based feeds present a more environmentally-sustainable alternative to fishmeal-based diets, but require further nutritional
optimization prior to commercial acceptance. Fishmeal substitution strategies have been used successfully in conjunction with
marine oils. However, when all marine fish oil sources are removed, shrimp performances declines due to a lack of essential
fatty acids. Previous experiments have demonstrated that supplementing plant oils with essential fatty acids can be done, but is
quite expensive. A more economical alternative is to minimize the levels of marine oils by diluting them with other oils such
as soybean oil. The objective of this study was to evaluate the response of shrimp to diets with decreasing levels of fish oil as
replaced by soybean oil.
A 10-week trial was conducted with Litopenaeus vannamei juveniles (1.37 ± 0.05 g), previously conditioned to a commercial
shrimp diet (35% CP, Rangen, Buhl, Idaho). Shrimp were stocked in a recirculating system consisting of forty 800 L HDPE
tanks (0.65 m 3 water volume) at a density of 26 shrimp/tank (40/m 3 or 31/m 2 ). Water was recirculated at 1.9 L/min/tank and
aeration was provided by two airstones (7-10 Lpm/stone). Naturally-induced primary production was present and no external
biofiltration was provided. The study evaluated a total of six diets with five replicates per treatment. Diets containing no fishmeal
and targeted crude protein levels of 35% were formulated with soy:fish oil ratios of (100:0, 50:50, 60:40, 70:30, 80:20,
and 90:10) and 6% lipid. A 35% CP diet (Rangen Inc., Buhl, Idaho) served as a reference diet. Feed was offered three times
daily. Rations were determined based on an assumed growth rate of 1.5 g/wk, an FCR of 1.4, and 100% survival. The study
was terminated when shrimp achieved a mean final weight above 20 g. This paper will discuss the results from this study.
231

232
SPOT Leiostomus xanthurus PRODUCTION ON EASTERN SHORE VIRGINIA FOR
MID-ATLANTIC BAITFISH MARKETS
Michael H. Schwarz*, Josh Reitsma, Brendan C. Delbos and Clarke Morton
Virginia Seafood Agricultural Research and Extension Center
102 South King Street
Hampton, VA 23669 USA
mschwarz@vt.edu
SKYPE: michaelhschwarz
Spot (Leiostomus xanthurus) are a highly desired baitfish in the Mid-Atlantic for a multitude of angling species including
striped bass, flounder, weakfish, and tuna. While wild-caught spot compete with aquacultured fish in the bait markets during
warmer months, winter fishing for stripers coincides with a lack of wild spot availability. This presents a unique production and
marketing opportunity for baitfish aquaculture of this species.
Biological and technical advancements towards culture of this species date back to the late 1970’s. In recent years, concerted
efforts by NOAA and Sea Grant accelerated research into marine baitfish production, culminating in VSG-05-08, A Virginia
Sea Grant publication entitled “Production of Spot as Live Bait for Recreational Angling.” These early efforts, in conjunction
with ongoing support from Virginia Sea Grant and Virginia Cooperative Extension assisted Mid-Atlantic Aquaculture
Technology in development of its spot production program for the baitfish market. This presentation will review development
of Mid-Atlantic Aquaculture Technology’s spot production program, production protocols for this species, and discuss market
development.

SHRIMP RESEARCH ACTIVITIES AT OCEANIC INSTITUTE
Shaun M. Moss * , Dustin R. Moss, Steve M. Arce, Clete A. Otoshi and Carrie M. Holl
Oceanic Institute
41-202 Kalanianaole Highway
Waimanalo, HI 96795 USA
smoss@oceanicinstitute.org
Through the U.S. Marine Shrimp Farming Program, Oceanic Institute (OI) has established a selective breeding program to
improve the performance of specific pathogen free (SPF), Pacific white shrimp, Litopenaeus vannamei. During the past year,
50 full-sib families from the TSV-Resistant line were produced. Representative shrimp from these families were exposed to a
single isolate of Taura syndrome virus (TSV; the Americas group) at the University of Arizona in a per os challenge test. After
14 days, mean family survival (± SD) was 88.7 ± 7.12% and family survival ranged from 70-100%. Twelve families exhibited
mean family survival > 95%, and 21 families exhibited mean family survival > 90%.
In addition to the TSV-challenge test, juvenile shrimp from all 50 families were evaluated for growth and survival in a 75-m 2
raceway located in OI’s Nucleus Breeding Center. For the trial, 8,604 tagged shrimp and 13,500 untagged shrimp were stocked
into the raceway resulting in an initial stocking density of 295 shrimp/m 2 . After 95 days, 7,283 tagged shrimp and 10,945 untagged
shrimp were harvested resulting in a final survival of 82.5%. Final harvest weight of the tagged shrimp was 23.38 g,
whereas final harvest weight of the untagged shrimp was 22.89 g. Mean weight of tagged, male shrimp was 22.54 g, whereas
mean weight of tagged, female shrimp was 24.25 g. These data provide further evidence of sexual growth dimorphism for this
species. Raceway production was 5.61 kg of shrimp/m 2 at harvest, Food Conversion Ratio (FCR) was 1.65, and water usage
was 1,206 L of water/kg of shrimp produced.
With regard to family-specific data, 11 families exhibited a mean family growth rate > 1.70 g/wk, and two families exhibited
a mean family growth rate > 1.80 g/wk. One family exhibited a growth rate of 1.86 g/wk. Mean family growth rate for all 50
families was 1.64 g/wk (SD ± 0.09). At the end of the growout trial, 28 families exhibited a mean family survival > 80%, and
two families exhibited a mean family survival > 90%. Mean family survival for all 50 families was 80.12% (SD + 08.74).
Correlation coefficients were estimated for TSV survival and Average Daily Growth, TSV survival and Growth Rate, TSV
survival and Production, and TSV survival and Inbreeding. Correlation coefficients ranged from 0.03 – 0.13 and none were
significant. These results suggest that there is no relationship between TSV survival and growth, as well as between TSV
survival and inbreeding, when inbreeding levels are low (F < 0.11).
233

234
EFFECTS OF SALINITY AND TEMPERATURE ON THE BLOOMS OF THE FISH KILLER
Cochlodinium polykrikoides ATSEPANGAR BAY, SABAH
Sitti Raehanah Muhamad Shaleh * , Joanna W. Doinsing, Ann Anton, Normawaty Mohammad Noor and Ejria Saleh
Borneo Marine Research Institute
Universiti Malaysia Sabah
Locked bag 2073
88999 Kota Kinabalu, Sabah, Malaysia
sittirae@ums.edu.my
Red tides or harmful algal bloom (HABs), a phenomenon that has caused serious environmental problems worldwide,
occurs frequently in the coastal waters of western Sabah, Borneo. Since the first occurrence in December 2004,
Cochlodinium polykrikoides, the species responsible for the reddish discoloration of surface waters at Sepanggar Bay, Kota
Kinabalu (N06°02’558’ and E116°06’397’), has resulted in mass mortalities of farmed fish in the University’s cage cultures
particularly the expensive groupers Epinephelus fuscoguttatus and E. lanceolatus. To elucidate the factors affecting the
abundance of C. polykrikoides, the physical seawater parameters; temperature and salinity during blooms and non-blooms
seasons were examined in-situ as well as laboratory experiments. Under controlled conditions, C. polykrikoides were grown
in F/2 medium and illuminated for 12 hours at 100 µmolm -2 s -1 . Different levels of salinity (5, 10, 15, 20, 25, 30, 35 and 40
ppt) and temperature (20, 25, 30 and 35 ºC) were tested. Field observation demonstrated two blooming seasons (> 5000 cellsL -
1 ), that are in June 2006 and January 2007.However, there were no significant correlation (p>0.05) between cell density and
both factors, the salinity and temperature. In laboratory, growth rate was significantly affected (P

THE IMPACT OF CATFISH IMPORTS ON THE U.S. CATFISH INDUSTRY
Andrew Muhammad, Sammy J. Neal and Terrill R. Hanson
Mississippi State University
Department of Agricultural Economics
Mississippi State, MS 39762 USA
muhammad@agecon.msstate.edu
The recent increase in catfish imports is of great concern to the U.S. catfish industry. The importing of tra, basa, and channel
catfish at relatively lower prices has resulted in less catfish purchased from U.S. farmers and processors. Claims have been
filed with the U.S. International Trade Commission (ITC) accusing Vietnamese exporters of selling catfish to the U.S. at less
than fair market value. Consequently, the U.S. International Trade Commission ruled against Vietnam in 2003 and recommended
tariffs from 37% to 64%.
The primary objective of this research is to assess the impact of the Vietnamese tariffs on the U.S. catfish industry. In this study,
we develop a supply and demand model of the U.S. catfish industry at the farm and wholesale level. In this model, we incorporate
the affects of imports and estimate the short-run and long-run effects of changes in import prices on U.S. prices, quantities
and welfare at the farm and wholesale level. Additionally, supply and demand elasticities are also estimated.
Preliminary Results
Tariff results are reported in Table 1. Average tariff rates 20.35%, 28.05% and 35.20% are considered. Without any tariff, U.S.
price and quantity at the wholesale level is $2.52/lb and 65 million lbs, respectively. Buyer and producer surplus are $114 and
$97 million, respectively. Revenue is $165 million. U.S. price and quantity at the farm level is $.90/lb and 145 million pounds
respectively. Buyer and producer surpluses are $27 and $121 million, respectively. Revenue is $138 million.
Given the maximum average tariff rate of 35.2% on imported catfish, U.S. wholesale price will increase by $.08 per pound and
quantity increases by 1.2 million lbs. Farm prices increase by $.02 per pound and farm quantity increases by 1.6 million lbs.
The changes in surplus (welfare) and revenue due to the tariff are also reported in Table 1.
Table 1. The Impacts Tariffs on The U.S. Catfish Industry
Vietnam
Tariff (%)
Price
($/lbs)
Quantity
(000 lbs)
Buyer
Surplus
($000)
Producer
Surplus
($000)
Revenue
($mill.)
Wholesale
Level
$2.52 65,427 $ 114,348 $97,449 165
20.35 +0.05 +690 +240 +3,910 +3
28.05 +0.07 +950 +348 +5,200 +4
35.20 +0.08 +1,200 +417 +6,420 +5
Farm
Level
$0.90 153,276 $27,810 $121,190 138
20.35 +0.02 +919 +335 +3,472 +4
28.05 +0.03 +1,260 +459 +4,763 +5
35.20 +0.04 +1,599 +584 +6,058 +7
23

23
FERTILIZATION OF OVOCYTES OF PIRACANJUBA (Brycon orbignyanus) USING
DIFFERENT VOLUMES OF WATER IN THE ACTIVATION OF THE SPERMATOZOIDS
Luis Murgas, Viviane de Oliveira Felizardo and Gilmara Junqueira M. Pereira
Veterinary Medicine Department
Federal University of Lavras
Lavras-MG, Brasil
lsmurgas@ufla.br
The piracanjuba (Brycon orbignyanus) is species of migratory spawning fish threatened by extinction in South America. The
present experiment was accomplished with the objective of evaluating the effects of the use of different volumes of water as an
activating solution for the spermatozoids on the rates of artificial fertilization of piracanjuba ovocytes. Four couples of selected
piracanjuba from the Itutinga Environmental Station reproducer tanks were used - MG (EAI-CEMIG), in the migratory spawning
period of 2007/2008. The animals received hormonal induction of crude carp hypophysis extract (CHE), for obtaining of
the gametes. The gametes were dry collected, via cranial-caudal massage of the animals. Ovocyte samples of 0.1 g from each
female were fertilized with 10µl of semen, using four different tank water doses (5, 10, 20 and 40 mL) for gamete activation,
doing two repetitions for each sample. After activation the eggs were placed in 1 liter experimental incubators, arranged inside
a 1000 liter tank, with constant water renewal, at an average temperature of 28ºC. After eight hours, the fertilization rate was
analyzed (viable eggs). The experiment was installed according to a random block layout with four treatments and two repetitions.
The averages were compared by the Tukey Test at 5%. The volumes of water tested didn't alter the fertility rates (P>0.05).
The use of 5 ml of water is recommended for activation of the spermatoids, so that there is practicality during the accomplishment
of the process.
ROLE OF SHELTERS ON PRODUCTION OF FRESHWATER PRAWN, Macrobrachium
rosenbergi
Shivananda Murthy, H.,* Kumarswamy, R. Shankar, K M Rajesh and K. J. Palaksha
Department of Aquaculture
Karnataka Veterinary, Animal and Fisheries Sciences University
College of Fisheries
Mangalore-575002, India
hsmurthy05@yahoo.com
Freshwater prawn, Macrobrachium rosenbergii is a commercially important species in India and other Asian countries.
Shelters play an important role in growth and survival of Macrobrachium rosenbergii, since it is aggressive and has territorial
and agonistic behavior. The effect of three types of shelters, namely, Eichhornea plant, used tires and PVC pipes on survival
and growth of Macrobrachium rosenbergii was investigated in soil based and fertilized cement tanks in triplicate. Tanks without
shelters were used as control . Growth of prawns in shelter based tanks was significantly (P < 0.05) higher than that of the
control. Relatively best growth and survival were achieved in tanks supplied with PVC pipes, which was followed by tyres
and Eichhornea plants. The percent increase in growth over that of the control was 168.22, 133.06 and 90.32 for PVC pipes,
tyres and Eichhornea respectively. Overall survival, net production and specific growth rate were superior in the PVC pipe
treatment. Growth and survival of Macrobrachium rosenbergii were increased by the introduction of shelters in culture tanks
and the magnitude of the effect appears to be dependent on the types of substrate. The increased growth and survival in general
in sheltered tanks is attributed to their use for refuge particularly for freshly moulted animals and increase in the surface area,
which reduce frequency of agonistic interactions.

EFFECTS OF TWO DIETARY LIPID LEVELS ON GROWTH PERFORMANCE AND BODY
COMPOSITION OF PRE-ADULT HATCHERY REARED AND WILD-CAUGHT BLACK SEA
BASS Centropristis striata
Md. Shah Alam, Amanda R. Myers*, Wade O. Watanabe, David Berlinsky, Abigail B. Walker, Katherine B. Sullivan
and Walker D. Wright-Moore
University of North Carolina Wilmington
Center for Marine Science
601, S. College Road
Wilmington, NC, 28403-5927 USA
alamm@uncw.edu
Two isonitrogenous (approximately 50% crude protein) formulated practical diets having different lipid levels (12% and 18%)
were fed to pre-adult black sea bass to test their effects on growth performance and body composition of hatchery reared and
wild-caught fish reared in pilot-scale recirculating sea water systems. In experiment 1, hatchery reared (approximately 395 g
initial weight) were fed the respective diets for about 6.5 months in triplicate recirculating tanks (2.7 m 3 , 50 fish/tank) at the
Center for Marine Science, University of North Carolina Wilmington (UNCW). In a separate experiment 2, wild-caught black
sea bass (approximately 177 g initial weight) were fed these same diets for about 4 months in triplicate recirculating tanks (4.8
m 3 , 10 fish/tank) at the Aquaculture Research Center, University of New Hampshire (UNH). In both experiments, fish were
fed two times a day to apparent satiation. At the end of the feeding trial, proximate composition for whole body, muscle, viscera
and liver tissue were analyzed.
In experiment 1, after 82 days of feeding, weight gain (g) and lipid content of whole body, muscle, liver and viscera for hatchery-reared
fish fed 18% lipid appeared to be higher than fish fed 12% but no significant differences were observed. After 193
days of feeding, hatchery reared fish fed 18% lipid showed significantly higher (P

23
MEASURING FEED INTAKE FOR CHANNEL CATFISH Ictalurus punctatus DURING COOL
WEATHER
Adam S. Nanninga and Carole Engle
Mail Slot 4912
University of Arkansas at Pine Bluff
1200 North University Drive
Pine Bluff, AR 71601 USA
ananninga@uaex.edu
Channel catfish Ictalurus punctatus fed based on standard winter feeding recommendations lost weight during winter feeding
studies conducted at the University of Arkansas at Pine Bluff. The objective of this study was to quantify feed consumption
by channel catfish over the winter months. Six concrete vats with a volume of 5 m 3 were stocked in mid October 2007 with 17
sub-market size fish (mean weight 0.25 kg; range of 0.11 to 0.39 kg) and 20 market size fish (mean weight 0.72 kg; range of
0.54 to 1.0 kg). Water was supplied by a pump on the bottom of an adjacent pond at 10.6 l/min. Feed (32% protein full-sink
pellet) was offered daily in feeding trays at a weight of 2% of total body weight of fish every afternoon. After 15 minutes, trays
were pulled up and remaining pellets were counted. All vats were harvested on 16 March 2008.
Channel catfish consumed from 0.1% to 0.7% of their body weight daily over the winter when offered feed every day. While
catfish ate more at higher water temperatures, they continued to eat at temperatures as low as 7 C. Compared with standard
winter feeding recommendations, daily feed consumed by catfish was slightly (0.2 - 0.3%) less than the recommended daily
percent of body weight to feed (at temperatures ≥ 10 C). However, the recommended frequency of feeding resulted in weekly
recommended feed quantities that were 1.3% - 1.6% lower than actual quantities of feed consumed by catfish. Additional work
is needed to refine feeding recommendations for cool-weather feeding, with particular attention to increasing the frequency of
feeding over the winter.
TABLE 1. Feed consumption by channel catfish during cool weather as compared
to standard winter feeding recommendations.
Standard Feeding Measured Feed
Temperature Range (C)
≤ 10 C
Recommendations Consumption
% body weight daily
0%
0.1%
% body weight weekly
> 10 – 15.6 C
0%
0.6%
% body weight daily
0.5%
% body weight weekly
0.5% a
0.3%
15.7 – 21.1 C
1.8%
% body weight daily
1%
% body weight weekly
3% b
0.7%
a
Feed only once a week.
b
Feed every other day.
4.6%

NOVEL COMPOUNDS AND OPPORTUNITIES FOR RENDERED ANIMAL PROTEINS IN
AQUACULTURE FEEDS
Sergio F. Nates, PhD. and Dominique P. Bureau, PhD
Fats and Proteins Research Foundation, Inc.
Alexandria, Virginia USA
Alternative sources for diets for aquaculture species are by-products produced by the inedible rendering industry, such as poultry
meal, blood meal, meat and bone meal, hydrolyzed feathers and many others. Rendered animal proteins are especially valuable
in the context of fishmeal replacement. Formulating low fish meal feeds requires the use of combinations of economical
ingredients since most feedstuffs have been shown to have significant limitations and cannot be used individually at very high
levels in the diet of most fish species. Rendered proteins are consequently very valuable for the formulation of cost-effective,
low fish meal aquaculture feeds. A large number of studies have shown that the quality of these ingredients has considerably
improved over the past two or three decades. Today, rendered animal proteins often prove to be the most cost-effective feedstuffs
when comparison is done on a level playing field, for example, on a digestible protein basis. The high nutrient density
(high protein, high fat, low carbohydrate) of “modern” shrimp and fish feeds leaves very little room for economical ingredients
with low protein and digestible energy contents (e.g., cereal grains, field pea, canola meal, soybean meal, etc.). The high
digestible protein and energy contents of most rendered animal protein ingredients makes them useful for the formulation of
low fishmeal, high nutrient density fish feeds.
Small peptides play numerous roles in the biology of aquatic species and hydrolyzed, rendered proteins seem to have opportunities
in the currently hot area of bioactive peptides. Numerous researchers have discovered that when certain proteins are
hydrolyzed in just the right way, the products will include one of these small peptides that have powerful physiological effects.
Recently, the research group of Chen (2007), a member of the Animal Co-Products Research and Education Center (ACREC),
a research center funded by the Fats and Proteins Research Foundation has attempted to produce bioactive peptides from rendered
protein such as collagen. In the preliminary research, some bioactive peptides have been screened and identified using
various chromatographic techniques and confirmed with different in vitro antioxidant bioassays. The solid data suggests that
one of these peptides could compete as a natural alternative to the popular synthetic antioxidants BHA and BHT, widely used
in the aquaculture feed manufacturing industry. In another study, we have quantified the nucleotide content of several rendered
animal protein ingredients. The idea is to identify how rendered animal proteins can be useful in improving the nucleotide
content of diets for first-feeding Atlantic salmon. Diets can then be used to enhance the growth and gut development of Atlantic
salmon larvae.
23

240
FAMILY COMPETITION IN A SELECTED STRAIN OF COHO SALMON (Oncorhynchus
kisutch)
Kathleen G. Neely, Jim Myers, Jeffrey J. Hard, Linda Park and Penny Swanson
National Marine Fisheries Service
Northwest Fisheries Science Center
Seattle, WA 98112 USA
kathleen.neely@noaa.gov
Effects of diet regime on between- and within-family competition was tested using a selected coho salmon strain (Dømsea
strain, Aquaseed, Inc). Fish from ten full-sib families were individually PIT tagged and reared in a communal tank and fed
at a limited ration (to allow study of competition effects) or, in a second tank, fed at twice the recommended ration (to allow
maximum growth and minimal competition). A previous study (Neely, unpubl. data) demonstrated that Domsea fish quickly
outperformed their progenitor population when fed to satiation. Furthermore, there was some evidence that feeding behavior
and competition had a strong influence on growth rate. This study investigated whether family differences in competition for
food influenced growth rate.
After one year of rearing, fish in the high ration group were 62% larger than fish in the low ration group. ANOVA indicated
that effects of both family and ration were highly significant (P>0.0001) throughout the study, with the exception of tank effects
at the initiation of the feeding study. This study proposes that within the range of ration treatments applied, the relative
performance of families was largely unaffected; however, suggesting that the efficacy of selection programs may be highly
dependent on the feeding regime.

HOLISTIC GOODNESS-OF-FIT: COMPARING OBSERVED AQUACULTURAL
PERFORMANCE WITH THAT SIMULATED VIA ECOPHYSIOLOGICAL MODELING
William H. Neill*, Robert R. Vega, Scott J. Walker and F. Michael Speed
Department of Wildlife and Fisheries Sciences
Texas A&M University
College Station, Texas USA
w-neill@tamu.edu
It is our contention that efficient advancement of aquacultural science requires a melding of statistical analysis and systems
modeling.
Most of us who are involved in aquacultural research find it expedient if not obligatory to make our measurements on systems
that are neither rigidly controlled nor exactly replicated—i.e., the data come from groups of animals in large tanks, ponds,
net-pens, and the like. The measurements themselves connect growth, survival and other performance outputs that tend to be
far removed, in a causal sense, from inputs such as environmental factors and feed treatments. Moreover, the lack of control
over the course of a 6- or 8-week-long study leads to uncertain interaction and physiological integration of time-varying inputs,
causing outputs that are inherently in transient-state. Even differential growth and survival of the animals themselves makes
for confounded relationships among intended treatments, unintended treatments (e.g., metabolite concentrations), and animal
size and biomass.
We think it’s time to move on, from statistical testing of trivial null hypotheses and uninformed description of associations and
relationships, to more thoughtful attempts at quantitative, mechanistic explanation of observed results. This requires that we
build informed models through which the data can be processed, then statistically test for consistency between modeled and
observed outcomes. Thus, all the complexity that stands between inputs and outputs becomes reduced to the issue of goodnessof-fit,
which is simply tested—only, there is one additional twist: Goodness-of-fit needs to be holistic, in that it must extend
across the various pairs of modeled and observed measures of performance, and over all available experiments. It isn’t enough
that growth be accurately modeled, if survival is not; and, it isn’t good enough if both “fits” are adequate in one instance but
not in another. There must be concordance overall, not just good fits here and there.
Lack of holistic goodness-of-fit provides both the motivation and guidance to re-think experiments, and, in the process, to gain
new insights about mechanisms and relationships. Often, those new insights can be formalized and then evaluated with the
same dataset—without running a new experiment.
We demonstrate our thesis by using Ecophys.Fish (Neill et al. 2004. Rev. Fish. Sci. 12:233-288) to model aquacultural performance
trials in tanks and ponds.
241

242
ECONOMIC CONSTRAINTS AND OPPORTUNITIES FOR MARINE BAITFISH
PRODUCTION IN LOW SALINITY WATERS OF THE ALABAMA BLACK BELT
Robert G. Nelson* and Ronald P. Phelps
Department of Agricultural Economics & Rural Sociology
Auburn University
Auburn, AL 36849 USA
nelsorg@auburn.edu
By taking advantage of saline groundwater and lower land prices, inland culture of marine baitfish is technically feasible in
many rural areas in the US, including the west Alabama Black Belt. However, a number of economic constraints have been
identified in the production and distribution of inland-cultured marine baitfish.
The principal baitfish species of interest currently are bullminnows (Fundulus grandis) and spot (Leiostomus xanthurus). Culture
of bullminnows appears to be financially attractive, and there is a well-established market along the Gulf Coast. Current
demand is thought to exceed two million bullminnows per year in Mississippi and Alabama alone, with shortages reported
regularly in the late summer. Total costs of production and transportation are estimated to be about $0.11/fish, and wholesale
market prices from auction sales have averaged $0.15/fish. Assuming average yields of about 173,000 fish/ha/yr, net revenues
of $6,920/ha are possible.
Bull minnows have been transported at densities of 120g/L for 5 h with over 95% survival from the inland production site to
the wholesale outlet at the coast. Bullminnows reproduced naturally over vegetation at 5 ppt in Black Belt ponds, but with this
extensive production system, stock management became an issue. All age groups were present in the pond and, with aquatic
vegetation serving as spawning material, harvesting by seining was unproductive. Trapping is currently our preferred method
of harvest, but many questions await experimentation in terms of trap design, placement, timing, bait, effort, efficiency, variability,
cost effectiveness, etc. Daily harvest by trapping has been widely variable, ranging from 2.5 to 41 kg/day. Also, the
uncertainty in estimating inventory in continuously-recruiting, partially-harvested ponds adds considerable complexity to the
optimization of yield. An alternative, but perhaps more labor intensive, system is mat-spawning in a brood pond, with removal
of mats to nursery/growout ponds, followed by seining at harvest. This system still needs to be field-tested.
Other candidate species of baitfish (spot, croaker, pinfish, and mullet) have to be induced-spawned. Culture systems for
these species usually involve recirculating water in indoor tanks, with separate broodstock, hatchery, nursery and growout
components. Fixed and variable costs are consequently much higher than pond-only systems, and enterprise feasibility has
so far proved to be unpromising. Nor do scale economies offer a simple solution to these constraints, since the pilot systems
demonstrated so far have generally proposed modular expansions. Possible alternatives include hybrid systems with indoor
broodstock and hatching, semi-intensive nurseries (with live feed harvested from ponds), and pond growout. Spot have proven
to be adaptable to the low-salinity conditions of west Alabama, but production costs associated with induced spawning make
their profit potential uncertain at this point.
Other constraints and opportunities involve holding and distribution systems. Complete pond harvests may yield more fish
than can be marketed at one time, thus requiring a holding facility. Likewise, if a bait species cannot survive in outdoor ponds
during the winter, or cannot be harvested from ponds in certain months, then a facility will be needed to hold large quantities
of fish indoors at high densities. These stocks could then be distributed from this facility to keep the market supplied continuously.
The design and cost of such a facility have yet to be determined.

DESIGNING A SUPPLY CHAIN FOR MARINE BAIT PRODUCED IN INLAND WATERS
Robert G. Nelson* and Ronald P. Phelps
Department of Agricultural Economics and Rural Sociology
Auburn University
Auburn, AL 36849 USA
nelsorg@auburn.edu
Baitfish culture in coastal areas is challenged by a number of formidable constraints. Land costs are prohibitively high due to
competition from vacation and retirement properties. Ponds dug in the sandy loam soils leak excessively and need to be lined
with clay or impermeable fabrics to retain water. The threat of hurricanes poses daunting financial risks from catastrophic crop
losses. However, all of these constraints are eliminated by growing marine baitfish far inland in ponds constructed in clay soils
and supplied with saline groundwater from certain widely-distributed aquifers. The technical feasibility of growing marine
baitfish using some saline groundwaters has been demonstrated, but commercial development is lagging.
This project seeks to use supply-chain management principles to coordinate the full spectrum of activities from collecting,
spawning and rearing several species of baitfish, to harvesting, assembling, hauling, holding, distributing, and selling to bait
dealers along the Gulf Coast. The main species being evaluated is the bullminnow (Fundulus grandis), although other candidate
species will be evaluated. The main activity – which makes this supply chain distinctively “demand-driven” – is regular
auction sales to retailers on a semi-weekly basis. All the other activities are programmed to support the marketing study, the
purpose of which is to produce reliable estimates of demand for baitfish species, sizes, quantities and qualities, as well as to
establish cooperative relationships among all the channel partners.
The conditions needed to grow large numbers of bullminnows necessitate additional research to move from demonstration to
commercial scale. Other species, such as spot, croaker, pinfish, and mullet, can also contribute to the enterprise. However,
they will first need to be collected, acclimated to ponds, and artificially spawned to complete their life cycle under culture
conditions.
For production from commercial-scale ponds to be efficient, researchers need to develop systems of inventory estimation and
control, methods for partial harvesting, and procedures for handling, holding and transporting fish in order to minimize stress
and mortality.
Finally, research needs to be directed to developing indoor recirculating systems to hold marine baitfish at high densities for
periodic retail deliveries along the Gulf Coast throughout the year.
243

244
CATFISH Clarias gariepinus FINGERLINGS AS BAIT FOR LAKE VICTORIA COMMERCIAL
FISHERIES
Charles Ngugi and Kwamena Quagrainie
Moi University
Eldoret, Kenya
cngugi@africaonline.co.ke
The artisanal fishery of Lake Victoria, Kenya has been degraded by environmental deterioration, and the stock of catfish in the
Lake has been drastically reduced because wild-caught catfish juveniles are used as bait for Nile perch hooks deployed daily
in the commercial fisheries industry. There is shortage of catfish bait for fishermen who need bait on a daily basis at an affordable
price to be able to continue fishing. Income opportunities therefore exist for current agricultural farmers to diversification
into small-pond aquaculture to provide farm-raised catfish fingerlings as alternative supply source of bait for the commercial
fisheries on Lake Victoria. The overall objective of the study was to increase catfish fingerling production as bait to reduce
overexploitation of indigenous species and conserve the diversity of Lake Victoria Fisheries. This intervention would transform
existing and potential small-scale fish farmers into high-yield profitable production groups through technical assistance in
propagation, production, general pond husbandry, and marketing.
Preliminary work in 2007 assisted in the formation of farmer clusters; all groups had been registered and opened operational
accounts. The Vihiga cluster specifically made far much progress than the other clusters and was able to design and implement
a comprehensive propagation programme that resulted into some experienced and new farmers realizing the full potential of
fingerling production. Sales for Vihiga cluster for the third quarter of 2007 showed that there was a progressive transformation
of baitfish farmers towards commercialization.
A marketing plan that included the current marketing situation, opportunities and threats, and a clearly defined marketing strategy
was developed taking into account market demographics, market trends, and market potential for growth. The strengths
and weaknesses of the farmer groups, baitfish traders, product offering, financing associated with the marketing channels were
analyzed. Results suggest that pricing of baitfish both at source and at end market is dependent on the supply and demand of the
baitfish. Demand is determined by the abundance of wild caught Nile perch from the lake. It was found that baitfish is sourced
from both the wild and farmed fish farmers. Bait traders are mainly women and operate in organized groups or as individuals
depending on the beaches along the Lake.
Some challenges faced included some farmers getting attached to food-size fish and unwilling to sell as bait; lack of funding to
purchase feeds and seed by farmers; predation on the farm; volatile baitfish price; and lack of quality feed for Clarias fry and
fingerlings.

ONTOGENY AND CHARACTERIZATION OF SOME INTESTINAL ENZYMES IN COBIA
Rachycentron canadum LARVAE
Nguyen Quang Huy, Helge Reinertsen, Tran Mai Thien and Elin Kjørsvik
Norwegian University of Science and Technology
N7491 Trondheim , Norway
huy.nguyen@bio.ntnu.no
The ontogeny of some intestinal enzymes were investigated during the development of cobia (Rachycentron canadum) larvae
in order to determine the timing of intestinal maturation. The larvae in one treatment was fed live feed only (LF) and in another
treatment was co-fed with a microdiet (L-MD) from 17 to 27 days post hatching (dph), and then microdiet alone to the end of
the experiment at 33 dph. The larvae grew exponentially from initial standard length (SL) 3.1 ± 0.02 mm at hatch to a final SL
20.6 ± 0.6 mm in LF and 21.3 ± 0.6 mm in L-MD treatment, with no significant difference between the treatments (p> 0.05).
However, survival rate at 30 dph of larvae in L-MD treatment (9.6%) was about almost half of that in LF treatment (19.8%). In
addition to intestinal enzymes, the pancreatic enzyme trypsin was assayed. Individual activity of trypsin increased with larval
growth as expected in both treatments. The specific activity of brush border membrane (bbm) alkaline phosphatase (AP) was
abruptly elevated from 23 to 26 dph (twofold; p< 0.05) in LF treatment and from 26 to 30 dph (2.5-fold; p 0.05) and about threefold between 26-30 dph in L-MD treatment (p< 0.05). No significant increase
in specific activity of bbm leucine aminopeptidase (LAP) and ratio of bbm LAP/leu-ala during larval development in both
treatments suggesting that LAP was a less clear marker for intestinal maturation than AP in cobia. As the activity of bbm AP
has been well recognized as intestinal maturation marker, it can be assumed that intestine of cobia larvae attained maturational
stage when bbm AP activity reached its plateau at 26 dph (SL 17.0 mm; 699.3 degree days) in LF treatment and at 30 dph (SL
20.0mm; 814.0 degree days) in L-MD treatment. Weaning of cobia larvae as early as 17 dph thus delayed the intestinal maturation
process clearly and was associated with lower survival rate than that of LF treatment, although growth rate of the larvae
was not affected.
24

24
A NEW PROCESS FOR PRODUCTION OF FISH MEAL AND OIL IN REMOTE AND
SEASONAL FISHERIES
Peter M. Nicklason*, Peter Stitzel, Ronald Johnson, Harold Barnett and Michael Rust
University of Idaho, Aquaculture Research Center
c/o Northwest Fisheries Science Center
2725 Montlake Blvd E
Seattle, WA 98112 USA
pete.nicklason@noaa.gov
A new mobile process incorporates stabilized
fish raw waste material to make valuable
co-products similar to conventional fishmeal
and oil. Stabilizing the fish allows volume control
and least capital design of the process facility.
Mobile construction of the process allows
the equipment to be moved between seasonal
fisheries to access additional raw materials.
The process described is being developed to
address the highly seasonal and geographically
dispersed Alaska salmon industry. Heads, trim,
and viscera from salmon canning represent
about 40% of the landed weight. The heads
and trim are processed using conventional fish
meal equipment to recover press cake and oil.
The press cake is blended with the uncooked
viscera and dried on a drum dryer to produce
a meal with minimal heating and drying time.
In periods of high or low volume, relative to
dryer capacity, material is stabilized with 0.8%
formic acid (w/w) and stored in tanks for later
drying to optimize dryer energy and capacity.
By concentrating, stablizing and storing the
fish raw material, sizing and operation of the
plant is more efficient. All the equipment for
this process can operate in shipping containers
and move using existing services in Alaska.
A feeding study using juvenile coho Salmon
(Oncorynchus kisutch) demonstrated comparable
growth on a diet of meal made from salmon
waste compared to a commercial grower diet.
Proximate and essential amino acid composition
are given in Table 1 for meal made from
whole spawned out salmon.
Table 1. Essential amino acids and Fig. 1. Feeding
study summary of proximate composition of experimental
experimental feed and commercial diet.
and control diets.
Experimental salmon Commercial
carcass feed control____
Arg 6.72 16.2 H2O 22.0
His 2.38 53.5 Prot. 46.2
Iso 4.08 17.0 Fat 14.8
Leu 7.02 13.3 Ash 10.5
Lys 7.49 0 CHO 6.5
Met 2.83
Phe 3.83
Thr 4.10
Try 1.01 Val 4.84
Essential Amino Acid
Arginine 6.72
Histidine 2.38
Isoleucine 4.08
Leucine 7.02
Lysine 7.49
Methionine2.83
Phenylalanine3.83
Threonine 4.10
Tryptophan 1.01
Valine 4.84

AMBIENT CONDITIONING AND ULTRASOUND ASSESSMENT OF FEMALE CHANNEL
CATFISH IN ARKANSAS AND LOUISIANA
Noel D. Novelo*, Jeff Baxter, Christopher C. Green, Robert P. Lang and Terrence R. Tiersch
Aquaculture Research Station
Louisiana State University Agricultural Center
Baton Rouge, LA, 70820 USA
nnovelo@agcenter.lsu.edu
Ultrasound imaging was evaluated in commercial-scale production of hybrid catfish (channel catfish Ictalurus punctatus x
blue catfish I. furcatus) in Arkansas and Louisiana. The goal of this study was to compare reproductive conditioning of female
channel catfish at ambient temperatures between two sites approximately 310 km apart (latitude difference of 2° 29´). The objectives
were to: 1) compare ambient water thermal profiles in broodstock holding ponds; 2) compare reproductive stages by ultrasound
imagery of females conditioned at ambient temperatures, and 3) evaluate induced spawning of females in Arkansas.
Mature (2.3 ± 0.5 kg) female channel catfish (N = 1,500) were transported from Arkansas (AR) to Louisiana (LA) in November
2007 for ambient conditioning during Spring. The LA ponds (0.3-ha) reached the average temperature threshold for gonad
recrudescence (21°C) 26 days earlier than AR ponds (2.6-ha) (Figure 1). Fish were collected, returned to AR, and selected for
induced spawning from April 29 to May 16, 2008 based on ultrasound data and by visual inspection of secondary sexual characteristics.
Gonad development stages were classified as: undeveloped (Stage 1); underdeveloped (Stage 2); developing (Stage
3); advanced (Stage 4); and mature (Stage 5). Gonad stage was significantly dependent (P < 0.05) on location. Gonads of 444
females (53% of LA females) were classified as Stages 4 and 5, and were selected for induced spawning at AR; of these, 61 ±
18 % spawned. Percent neurulation of egg masses at 28 to 30 h (n = 180; 300 to 400 ml eggs) was 33 ± 22 %. Gonads of 129
females (11% of AR females) were classified as Stages 4 and 5; of these, 82 females were selected for induced spawning, and
50% spawned. Percent neurulation of egg masses (n = 33; 300 to 400 ml eggs) was 30 ± 16 %.
Figure 1. The threshold temperature (21 C) for
gonadal recrudescence was reached 26 d earlier
in LA (solid line) than in AR (dotted line).
47%
28% 25%
5% 5%
24
Figure 2. Ovarian stages (mean ± SD) in LA
(black bars) and AR (white bars). There were
significantly more females (P < 0.05) at Stages
4 and 5 in LA and more in Stages 1 to 3 in AR.
Percentages are in relation to total sample size of
each location.

24
ACUTE TOXICITY OF 5% PRENTOX® PRENFISH ROTENONE TO FRESHWATER
PRAWN Macrobrachium rosenbergii
Tiffany Ogunsanya*, Robert Durborow, Carl Webster, James Tidwell, Hank Jarboe, Changzheng Wang,
Lingyu Huang and Yolanda Brady
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
Tiffany.Ogusnanya@kysu.edu
Predator fish, such as bluegill Lepomis macrochirus and green sunfish Lepomis cyanellus, are notorious for preying on juvenile
prawn Macrobrachium rosenbergii. Before the prawns are stocked, ponds are often drained in an attempt to kill “trash” fish
species; however, some species survive. Rotenone is one of the most frequently used piscicides to eradicate bluegill and green
sunfish. However, there is no data regarding the concentration of rotenone that is lethal to the prawns.
The primary goal of this study was to evaluate the acute toxicity of rotenone on juvenile freshwater prawns and to provide toxicity
data that is applicable for prawn farmers. The mortality response of juvenile prawns exposed to different rotenone concentrations
was evaluated during two 96-hour bioassay experiments to determine the LC50 values. For trial 1, freshwater prawns
were exposed to the following Prentox® Prenfish rotenone treatments; 0.0 mg/L, 1.0 mg/L (0.05 mg/L active ingredient),
3.0 mg/L (0.15 mg/L active ingredient), 5.0 mg/L (0.25 mg/L active ingredient), and 10.0 mg/L (0.50 mg/L active ingredient),
respectively. In the second 96-hour bioassay, M. rosenbergii were exposed to treatments of Prentox® Prenfish ranging from
0.0 mg/L, 2.2 mg/L (0.11 mg/L active ingredient), 3.6 mg/L (0.18 mg/L active ingredient), 6.0 mg/L (0.30 mg/L active ingredient),
10.0 mg/L (0.50 mg/L active ingredient), and 16.7 mg/L (0.84 mg/L active ingredient), respectively.
Results from trial 1 showed that juvenile prawns were able to achieve 100% survival when exposed to Prentox® Prenfish
rotenone concentrations of 1.0 and 3.0 mg/L, while mortalities did occur at the 5.0 and 10.0 mg/L treatments (Table 1). Results
from this study provide new insights on rotenone tolerances in juvenile freshwater prawns and may improve the culture
practices for this species.
Table 1. Survival of juvenile prawns during the 96-h toxicity Trial 1.
Prentox® Prenfish
rotenone treatment
Number of prawns per treatment Survival of prawn
1.0 mg/L (0.05 AI) 24 100%
3.0 mg/L (0.15 AI) 24 100%
5.0 mg/L (0.25 AI) 24 54%
10.0 mg/L (0.50 AI) 24 21%

AQUACULTURE SAFETY IN KENTUCKY
Tiffany Ogunsanya*, Henry Cole, Robert Durborow and Mel Myers
Aquaculture Division
Kentucky State University
Frankfort, KY 40601 USA
Tiffany.Ogunsanya@kysu.edu
An aquaculture safety survey resulted in finding safety ideas implemented by private and government fish production facilities.
Safety precautions are also pointed out in this study. Findings include the following points that may help aquaculturists reduce
accidental injuries by creating a safer working environment:
• The use of pulleys reduces muscle strain when lifting fish in nets.
• Using pepper plate on hauling truck steps provides traction even in wet conditions.
• Slippery pond liners sometimes prevent workers from exiting ponds; ropes extending into the pond give workers something
to grab onto to pull them from the pond.
• Salt should be kept off of floors to prevent slipping (salt crystals can act as small ball bearings).
• Lifting smaller weights of fish in dip nets (thereby working faster with smaller weights) resulted in higher work output
compared to making fewer trips with heavier quantities of fish.
• Removable guard rails on truck beds hold cargo in place when inserted but when removed, they make loading operations
safer for workers boarding and exiting the truck bed.
• Using shutes to unload fish can avoid back strain.
• Caution should be taken when working on narrow concrete raceway walls at trout farms.
• Debris should be kept off of raceway walls to avoid tripping and falling.
• Safety shields should be used over PTOs.
• Electricity is commonly turned off at breaker boxes while doing electrical work; however, workers need to be aware of
connecting, adjacent breaker boxes that kick on automatically after the main source has been off for several minutes.
• Rotting wooden piers on ponds can collapse and injure pond workers.
• Raised berms on the edge of cliff side roads can prevent vehicles from rolling off the cliff.
24

2 0
CLONING, EXPRESSION AND FUNCTIONAL CHRACTERIZATION OF AGARASE FROM
MARINE BACTERIA STRAIN AG4
Chulhong Oh, Youngduk Lee, Ilson Whang, Ji-Hee Lim, Se Jae Kim and Jehee Lee
Department of Marine Biotechnology
College of Ocean Science
Cheju National University
Jeju, 690-756, Republic of Korea
Agar is a polysaccharide which linked D-galactose and L-3,6-anhydro-galactose residues. The ability of degradation of agar or
agarose to oligosaccharides makes it widely used in food, cosmetic and medical industries. Agarase is produced mainly by bacteria.
Alpha-agarase cleave alpha-1,3-anhydro-L-galactosidic bonds and beta-agarase cleave the beta-1,4-galactosidic bonds. In
recent years, studies have demonstrated that the oligomer forms derive from agar or agarose exhibits variety physiological and
biological activities. The extent of these activities is also correlated with the degree of polymerization and the galactosyl groups
on the AOS and NAOS. Neoagarobiose (N2) was reported to process moisturizing effect on skin and and whitening effects on
melanoma cells (Kobayashi et al. 1997). Neoagarotetrose (N4), derived from prophyran, was reported to be utilize in vitro by
intestinal bacteria, which stimulated the growth of Bacteroides, as well as Eubacterium and Lactobacillus.
In this study, we identified agarase gene by PCR and sequencing from agarase producing marine bacteria strain. The size was
870 bp (290 aa) including 63 bp (21 aa) signal sequence and only 2 bp (2 aa) different compare to Pseudoalteromonas sp. KJ
2-4 alpha-agarase gene. Molecular weight and isoelectric point was 33 kDa and 5.9 respectively. The sequence showed GH16_
beta agarase domain. The mature sequence was cloned into pMal c2x expression vector and transformed to E. coli BL21 (DE3).
Finally the maltose binding protein-agarase fusion protein expression was induced by adding 0.5 mM IPTG. Inducted cell was
lysised by ultrasonicator and divided soluble and insoluble protein by centrifugation. Expression level was checked by SDS-
PAGE and activity test. Recombinant protein was purified with pMAL protein fusion and purification system. Characterization
and analysis of purified protein was carried out for optimum temperature, optimum pH and teen layer chromatography. The
optimum temperature and pH were 45 ℃ and 6.0. We made neoagaro-oligosaccharide with purified agarase. The product
showed activity about anti-oxidant and whitening effect.

SUCCESSFUL CULTURE OF PINFISH Lagodon rhomboides AND PIGFISH Orthopristis
chrysoptera IN RECIRCULATING AQUACULTURE SYSTEMS AT VARIOUS STOCKING
DENSITIES
Cortney L. Ohs*, Scott W. Grabe, Shawn M. DeSantis, Andrew L. Rhyne and Matthew A. DiMaggio
University of Florida
School of Forest Resources and Conservation
Program in Fisheries and Aquatic Sciences
Indian River Research and Education Center
2199 S. Rock Road
Fort Pierce, FL 34945 USA
cohs@ufl.edu
The culture of marine baitfish has great potential in coastal states. Almost all marine baitfish used by anglers are collected
from the wild and are subject to seasonal availability of species and demanded sizes of fish. Little research has been conducted
on the culture of pinfish and pigfish even though they are popular bait species in the Gulf and southern Atlantic coastal states.
Therefore, experiments were conducted to determine the effects of various stocking densities on growth and survival of pinfish
and pigfish in recirculating aquaculture systems.
Pinfish obtained from the wild were stocked at four densities (0.5, 0.2, 0.4, 0.6/L) at an initial mean weight of 5.36 g and mean
length of 8.9 cm into three separate recirculating aquaculture systems. Each system had four tanks containing 800 L of natural
seawater and one replicate of each treatment was randomly assigned to each system. Each system consisted of a bead filter,
foam fractionator, and a UV sterilizer. Throughout the experiment salinity was maintained at 28±2 g/L. Pinfish were fed a
commercially available diet for 82 days. At the conclusion of the experiments survival, growth, and FCR was determined.
Treatment survival means ranged from 94.3 to 99.2% with no significant differences between treatments.
Pigfish obtained from the wild were stocked at three densities (0.1, 0.3, 0.5/L). Mean weight of fish in each tank ranged from
8.6 to 11.6 g and mean length ranged from 8.4 to 9.6 cm. The system consisted of twelve 85 L aquaria equipped with a biofilter,
foam fractionator, and UV sterilizer. Each treatment density was replicated four times. Salinity throughout the experiment was
28±2 g/L. Pinfish were fed a commercially available diet for 65 days. At the conclusion of the experiments survival, growth,
and FCR was determined. Treatment survival means were 22% for 0.1/L, 38.3% for 0.3/L, and 57.8% for 0.5/L. This lower
than desired survival may have been due to the smaller size of tanks used or the aggressive nature of pigfish.
Both pinfish and pigfish show great promise as new aquaculture species and they have well established markets with high
demand and seasonal availability.
2 1

2 2
EVALUATION OF SPAWNING SUBSTRATE PREFERENCE OF Fundulus seminolis
CULTURED IN TANKS
Cortney L. Ohs*, Scott W. Grabe, Shawn M. DeSantis and Matthew A. DiMaggio
University of Florida
School of Forest Resources and Conservation
Program in Fisheries and Aquatic Sciences
Indian River Research and Education Center
2199 S. Rock Road
Fort Pierce, FL 34945 USA
cohs@ufl.edu
The seminole killifish, Fundulus seminolis, is an endemic Florida freshwater killifish that has emerged as a potential candidate
for marine baitfish aquaculture following successful saltwater acclimation. Its ability to complete its reproductive life cycle in
freshwater is appealing for aquaculture in Florida as value of coastal property increases and access to seawater becomes more
limited. It is unknown if F. seminolis is capable of substrate spawning and air incubation in cool moist conditions like other
Fundulus spp.
Therefore, a series of experiments were conducted to determine the preferred type, location, and orientation of various substrate
materials for spawning by F. seminolis in outdoor flow through tanks. Substrates evaluated included trays of sand and spawntex
mats. Additionally, three orientations were evaluated for spawntex mats including horizontal on the bottom, horizontal in the
water column, and vertical against the tank wall. In each experiment each substrate type was replicated four times. Some tanks
had each substrate type simultaneously presented and other tanks had each substrate type individually presented to fish for
potential spawning. The position of each substrate type in each tank was rotated in the four replicates to control for a potential
positional effect within a tank. Twelve 900 L rectangular tanks were used and each was provided with flow through water (1
g/L salinity) from a lined pond. Each tank was stocked with spawning populations of 4 males and 8 females. Once weekly the
substrates were removed and the number of eggs was visually quantified.
F. seminolis did not use the trays of sand for spawning even when this was the only substrate type presented to them. F. seminolis
used spawntex mats in both vertical and horizontal orientation extensively for spawning. There was no clear preference
when both vertical and horizontal orientations were presented simultaneously. Finally, evaluations of air incubation of eggs
confirm under cool and moist conditions F. seminolis eggs can be maintained out of water for up to 10 days while embryos
continue to develop. Upon re-submersing the embryos into water, hatching is immediate.

PRELIMINARY STUDY OF THE EFFECTS OF ELEVATED DIETARY CADMIUM ON
GROWTH AND CADMIUM ACCUMULATION IN TISSUES OF JUVENILE PARROT FISH,
Oplegnathus fasciatus
Okorie E. Okorie, Jun-Ho Lee, Seunghyung Lee, Young Chul Kim, Jun-Young Bae, Gun Huyn Park
and Sungchul C. Bai*
Department of Aquaculture/Feeds and Foods Nutrition Research Center
Pukyong National University
Busan 608-737, Republic of Korea
scbai@pknu.ac.kr
An 8-week feeding trial is going on to evaluate the effects of elevated dietary cadmium on growth and cadmium accumulation
in tissues of juvenile parrot fish, Oplegnathus fasciatus, averaging 5.5±0.06 g (mean±SD). Seven semi-purified experimental
diets were formulated to contain 0, 100, 200, 400, 800, 1600 and 3200 mg Cd/kg diet in the form of cadmium chloride (CdCl 2 ).
Casein and white fish meal served as the protein sources, wheat flour, dextrin, and corn starch as the carbohydrate sources, and
fish oil as the lipid source of the experimental diets, which were isonitrogenous and isoenergetic, and contained 50% crude
protein and 17 kJ available energy/gram diet. After two weeks of acclimation, fifteen parrot fish were randomly distributed into
each of 21 acryl tanks of capacity 30 liters. Each diet is being fed to triplicate groups of fish twice a day (1000 and 1600 h) to
apparent satiation (approximately 4% of wet body weight per day at the beginning and planned to be reduced to 3% of wet body
weight per day at the end of the feeding trial). At the end of 8 weeks of feeding trial, weight gain (WG), specific growth rate
(SGR), feed efficiency (FE), protein efficiency ratio (PER), and survival will be measured. Hematological characteristics to be
determined include hematocrit (packed cell volume – PCV) and hemoglobin, Hb (g/dl). Cadmium accumulation in gill, liver,
muscle and kidney will be determined.
2 3

2 4
TESTING FEED RESPONSE METHODS OF INVENTORY ESTIMATION FOR CHANNEL
CATFISH Ictalurus punctatus PRODUCTION
Pratheesh Omana Sudhakaran, David Heikes and Carole Engle
Department of Aquaculture/Fisheries
University of Arkansas at Pine Bluff
Pine Bluff, Arkansas 71601 USA
pomana@uaex.edu
The multiple batch production system commonly used in commercial catfish production results in a mixture of sizes of fish
in the pond. Accurate, cost-effective inventory estimation would improve financial and production management. Several inventory
estimation methods used by industry consultants and advisors are based on feeding response of fish. Daily feeding
response is known to be variable; hence, biomass estimates based on daily feed response would be expected to be variable. No
systematic tests of the variability and accuracy of these methods have been conducted. Datasets from experimental trials of
catfish production in single and multiple batch systems at various densities were used to estimate and compare: 1) the variation
in feeding rates, and 2) compare actual catfish biomass with biomass estimated with five different methods. Coefficients
of variation (CV) of quantity of feed fed per treatment were calculated for each week for each pond within three temperature
ranges. Analysis of variance (ANOVA) was used to compare CVs calculated using daily feed, two-day means, three-day means
and four-day means for each temperature range, for each stocking density and feeding treatment, for both single and multiple
batch. Stocking density and temperature had no significant effect on variation of feeding rates in either single or multiple batch
production. However, in single batch production, the four-day average of daily feed showed a significantly lower CV (17%)
than daily feed (mean CV= 38%), two-day means (mean CV= 27%) or three-day means (mean CV= 24%) (Figure 1a). In
multiple batch production, the three-day average of daily feed had a significantly lower CV (24%) compared to the daily feed
(mean CV= 49%) and 2-day average (mean CV= 34%) but was not significantly different from the four-day average (mean
CV= 23%) (Figure1b). Multiple batch feeding quantities were generally more variable than those in single batch. Actual catfish
biomass will be compared with inventories estimated with five different feed response methods utilized by industry advisors.
An inventory estimation error will be calculated for each.

METHYLMERCURY CONCENTRATIONS FOUND IN WILD AND FARM RAISED
PADDLEFISH IN KENTUCKY
Richard J. Onders, Siddhartha Dasgupta, Steven D. Mims and Deke T. Gundersen
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
rick.onders@kysu.edu
Kentucky is one of 18 states with statewide fish consumption advisories due to mercury contamination. These advisories are
renewed annually; however, they are based on limited and intermittent data. In addition, a more restrictive advisory is issued
for paddlefish consumption. We compared mercury levels in the muscle tissue of paddlefish from two wild sources: Ohio River
and Lake Cumberland, and from two farmed sources (polyculture with catfish and reservoir ranching) with the advisory levels.
We also compared these levels with those typically present in species popular with consumers, both freshwater and marine.
The results will be presented.
RESERVOIR RANCHING OF PADDLEFISH Polyodon spathula
Richard J. Onders and Steven D. Mims
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
rick.onders@kysu.edu
Reservoir ranching is a sustainable extensive fish production method that is practiced worldwide.
It is based upon artificial propagation and stocking of the desired species, with grow out entirely dependent on the natural productivity
of the reservoir. Therefore, it is best suited to species which feed low on the food chain and have potential for rapid
growth. The paddlefish Polyodon spathula is such a species as it is a zooplanktivore that is known to grow rapidly. In addition,
paddlefish roe is valuable as a source of black caviar.
In the US, there is a history of commercial fishing in large reservoirs, primarily for such species as catfish Ictalurus spp.,
buffalo Ictiobus spp. and common carp Cyprinus carpio. However, state agencies have restricted these fisheries over several
decades because of the perception that they interfere with recreational fishing. This has produced a culture of opposition to
reservoir ranching in the US, as stocking fish for commercial harvest would reverse the decline of inland fisheries, and this
would be viewed as undesirable by recreational fishing groups and state fishery managers.
In 2006, under pressure from state legislators, the Kentucky Department of Fish and Wildlife Resources enacted regulations
allowing reservoir ranching of paddlefish in qualifying municipal water supply reservoirs. Developments before and since
enactment of these regulations will be reviewed.
2

2
SALINITY AND TEMPERATURE EFFECTS ON GROWTH, SURVIVAL, WHOLE-BODY
OSMOLALITY, AND MRNA EXPRESSION OF NA + , K + , ATPASE ON LARVAL RED PORGY
Pagrus pagrus
Andrew D. Ostrowski*, Frank A. Montgomery, Troy C. Rezek, Thomas H. Shafer, James A. Morris Jr. and Wade O.
Watanabe
University of North Carolina Wilmington
Center for Marine Science
601 S. College Rd.
Wilmington, NC, 28403-5927 USA
ado2046@uncw.edu
Atlantic Red porgy, Pagrus pagrus, is an important demersal finfish species in the snapper-grouper complex off the southeastern
United States. Stocks have declined severely due to overfishing even with management measures in place during the past
decade. In the Mediterranean, and more recently in the U.S., red porgy has been found to be a viable candidate for aquaculture
with its high market value and its ability to spawn freely in captivity. Optimal rearing conditions for larviculture are currently
being developed at the UNCW. This paper examines the combined effects of temperature and salinity on eggs, yolk-sac larvae,
and early feeding-stage larvae of red porgy.
To determine the optimal temperature and salinity conditions for culture from hatching to day 16 post-hatching (d16ph), embryos
were stocked into black 15-L tanks at 60 L -1 under four temperatures (17, 19, 21, and 23ºC) and two salinities (24 and 34 g
L -1 ) in a 4 x 2 factorial design. Light intensity was 500 lx, aeration was 90 mL min -1 , and photoperiod was 16 L:8 D. Significant
(P < 0.05) effects of temperature and salinity on growth (notochord length, wet and dry weight), survival, whole-body osmolality,
and mRNA expression of Na + , K + , ATPase were observed with minimal interactive effects. Under both salinities, growth
increased with increasing temperature. On d16ph, wet weight (mg) at 21 and 23 ºC (2.03 and 2.91) was significantly (P < 0.05)
higher than at 17 and 19 ºC (0.20 and 0.69). Under all temperatures, salinity had no effect on growth but had a significant (P
< 0.05) effect on survival to d16ph, with greater survival at 24 g L -1 (18.4%) than at 34 g L -1 (6.77%). Salinity significantly (P
< 0.05) affected whole body osmolality (mOsm kg -1 ) on d2ph (342 in 24 g L -1 vs. 522 in 34 g L -1 ) and on d17ph (412 vs. 481).
Except for d6ph, when whole body osmolality at 23 ºC (401) was significantly lower than at 21 ºC (683), no clear temperature
related effects were observed throughout the study. On d16ph, when all larvae were challenged with a sublethal salinity increase
to 44 g L -1 , larvae from 34 g L -1 treatments did not show an increase in mRNA expression of Na + , K + , ATPase after 24
or 48 h post transfer. However, fish in the 24 g L -1 treatments showed a significant (P < 0.05) increase in mRNA expression of
Na + , K + , ATPase after 24 h followed by a decrease after 48 h. Within the ranges tested, a temperature of 23 ºC and a salinity of
24 g L -1 are optimal for culture of red porgy embryos, yolk-sac, and first-feeding stage larvae to d16ph.

COMPARATIVE OXYGEN TOLERANCE OF BLUE CATFISH Ictalurus furcatus AND
CHANNEL CATFISH I. punctatus
Les Torrans* and Brian Ott
USDA ARS Catfish Genetics Research Unit
Thad Cochran National Warmwater Aquaculture Center
Stoneville, MS 38776 USA
Les.torrans@ars.usda.gov
Blue catfish (Ictalurus furcatus) have several advantages over channel catfish (I. punctatus) as commercial culture fish. They
are more resistant to ESC, PGD, and CCVD, major diseases of channel catfish. They are very easy to seine, with a near-total
harvest possible with one seine haul, and may exhibit more uniform growth. They have a greater carcass yield (headed, gutted,
skinned) than channel catfish, although they do yield a smaller shank filet. Perhaps the biggest perceived disadvantage is the
reported poorer tolerance of low dissolved oxygen (DO) by blue catfish; however, this assessment has been based largely on
anecdotal evidence.
Studies were conducted in 15 1-acre and/or six ¼-acre ponds over four different years to determine the effect of low DO concentration
on the feed consumption and other production parameters of blue catfish (BC) compared to channel catfish (CC).
The oxygen treatments were based on the mean minimum DO concentration during the growing season (June-September).
Control ponds in each study were maintained with a minimum DO concentration above 4.3 mg/L; test ponds were maintained
at 1.6 – 3.0 mg/L in different studies (see figure). The fish were fed once daily to apparent satiation in all studies. Ponds were
clean harvested at the end of the growing season.
The results indicate that low DO has less of an impact on the feed consumption and other production parameters of BC than
CC. At a mean minimum DO concentration of 1.7 mg/L, feed consumption of CC was reduced by 45%; at 1.6-1.7 mg/L, feed
consumption of BC was reduced by only 21%. Behavior may play a larger role than physiology in the perceived differences in
the species. The data indicates that blue catfish may have more potential as a commercial food fish than previously thought.
2

2
ROUTINE METABOLIC RATE AND LIMITING OXYGEN CONCENTRATION OF
FRESHWATER PRAWN Macrobrachium rosenbergii LARVAE
Brian D. Ott* and Eugene L. Torrans.
USDA ARS Catfish Genetics Research Unit
Thad Cochran National Warmwater Aquaculture Center
P.O. Box 38
Stoneville, MS 38776 USA
brian.ott@ars.usda.gov
Malaysian prawns, Macrobrachium rosenbergii, are hatched and raised indoors in small tanks. Prawns may be raised and
shipped at high densities which could result in low dissolved oxygen (DO) conditions. Because DO may play an important role
in prawn development and survival, we measured routine metabolic rate (RMR) and limiting oxygen concentration (P crit ; the
presumed hypometabolic transition zone) of prawns at four different ages.
Measurements were taken in a closed respirometer consisting of a 300mL BOD bottle fitted with a Hach BOD overflow funnel.
Oxygen consumption ( V & o 2 ) and temperature were measured using a Hach HQ10 luminescent DO meter equipped with a spin
bar on the tip of the probe. Prawn larvae were collected either from hatching or rearing troughs at Lauren Farms, Inc., Leland,
MS. V & o 2 was measured initially after they hatched (0 days post hatch [dph]), at 15 dph and 29 dph, and immediately after the
prawns transformed into post-larvae (31 dph). For each measurement, a prawn sample was collected in a dip net, patted dry and
weighed (1.0 – 1.2g, n=6,000-120), and placed in a respirometer where DO and temperature were measured every minute for
30-100 minutes at an average water temperature of 26.0°C. The P crit was determined as the DO concentration at which prawn
V & o 2 decreased from a linear rate.
Individual prawn V & o 2 was lowest at 0 dph (0.00023 ± 0.00003 mg O 2 n -1 h -1 ) and was 11-, 30-, and 49-fold higher than intial
rates at 15 dph, 29 dph, and 31 dph, respectively. Mass-specific V & o 2 remained unchanged over the four sampling periods and
ranged from 1216 ± 144 mg O 2 kg -1 h -1 on 0 dph to 1300 ± 49 mg O 2 kg -1 h -1 on 31 dph. Limiting O 2 concentration was lowest
at 0 dph (53 ± 1% saturation), peaked at 29 dph (74 ± 1%), and decreased once they became post-larvae (69 ± 1%). Oxygen
consumption and limiting O 2 concentration are factors to consider when raising and shipping prawns and become more critical
issues as they age and grow.

A STANDARD GENETIC STOCK OF RAINBOW TROUT FOR RESEARCH STUDIES
Ken Overturf* and Ronald Hardy
USDA-ARS
Hagerman Fish Culture Experiment Station
3059-F National Fish Hatchery Road
Hagerman, Idaho 83332 USA
Ken.Overturf@ars.usda.gov
Research studies comparing similar diets, ingredient substitution or effects of nutrient supplementation often exhibit significant
differences in the response of fish compared to results from similar studies. Furthermore, researchers attempting to replicate
specific physiological signs associated with antinutritional factors in certain feed ingredients found by other researchers are
often unable to duplicate the original findings under their laboratory conditions. There are many explanations given for such
different findings, including differences in rearing conditions, such as water temperatures and flows, differences in feeding
level or frequency, or method of data analysis. However, in published discussions from several of these studies the authors suggest
that the differences reported in their findings, when compared to what others had found, could be do to genetic differences
between stocks of fish. Most rainbow stocks used in research studies come from eggs purchased from commercial suppliers,
local broodstock, academic stocks, or wildlife management facilities. One solution to this potential problem is to make eggs
from a single, defined stock of rainbow trout available to researchers. Through funding made available from the US Soybean
Board, the University of Idaho and USDA/ARS are generating a research stock of rainbow trout whose makeup will consist
of a cross between two commercial strains. The parent strains for this stock will be subjected to photoperiod manipulation so
that fish can be spawned at up to four different times per year, making eggs available to researchers throughout the year. Initial
stocks were received in the spring of 2007 and the first crosses will be generated beginning the fall of 2009. These stocks of
fish should prove a valuable resource for researchers by limiting differences detected in studies thought to be due to variation
in the genetic background of rainbow trout used in feeding trials
2

2 0
EXPRESSION OF REGULATORY ENZYME GENES IN RELATION TO LIPID UTILIZATION
AND DEPOSITION IN RAINBOW TROUT
Ken Overturf*, T. Gibson Gaylord and Frederick Barrows
USDA-ARS
Hagerman Fish Culture Experiment Station
3059-F National Fish Hatchery Road
Hagerman, Idaho 83332 USA
Ken.Overturf@ars.usda.gov
In developing a sustainable diet for piscivorous fish species it will be necessary to not only substantially reduce the current
level of fishmeal, but also the level of fish oil. Standard salmonid diets use between 9 and 15% fish oil which functions as
a key source of energy and essential fatty acids. While aquaculture is currently using around 50% of the fishmeal produced
worldwide it is using greater than 80% of the produced fish oil. In an attempt to replace fish oil studies have been carried out
to evaluate the potential of using oils obtained from sustainable sources such as plants. Research has determined that most of
the fish oil currently used in salmonid diets can be replaced with plant derived oils without compromising growth performance.
However, there is a substantial change in the fatty acid composition of the fillets in fish reared on these diets that generally
mimic the fatty acid profile of the dietary lipids. Studies using phase feeding or finishing diets containing fish oil have shown
an increase in the level of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in fillets over that of fish fed diets
containing only plant oils.
In previous studies evaluating the effect of fishmeal and fish oil replacement diets on spawning fish and egg quality, a significant
difference was observed in the level of expression of a key enzyme involved in regulating fatty acid biosynthesis. In this
experiment expression of the enzyme Δ6 desaturase was determined to be upregulated in fish fed a diet that had fishmeal and
fish oil replaced with plant protein and oil. Furthermore, it was noticed that this upregulation was significantly higher in some
families on all diets and that difference even more pronounced when the fish were reared on the formulated plant meal and
plant oil replacement diet (Fig 1). To determine the potential level of variation within rainbow trout, thirty-five families of fish
were reared for 12 weeks on a diet formulated with plant based fatty acid precursors for the synthesis of EPA and DHA. At the
end of the study an average of 25 fish from each family was sampled to determine the deposition of fatty acid in the fish and to
evaluate the expression of mRNA for three genes involved in the biosynthetic pathway, namely Δ6 desaturase, Δ5 desaturase,
elongase, and CoA dehydrogenase. Final analysis is currently underway with these samples.

ADVANCES IN THE DEVELOPMENT OF THE GERMPLASM BANK OF AQUATIC SPECIES
OF BAJA CALIFORNIA MEXICO
Carmen G. Paniagua-Chavez
Department of Aquaculture
Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE)
Kilómetro 107 Carrt. Tijuana-Ensenada
Ensenada, Baja California, 22800, México
cpaniagu@cicese.mx
In 2005, the Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE) launched the first Germplasm
Bank of Aquatic Species (GBAS) of Baja California. This project was developed with funds of the National Council of Science
and Technology (CONACyT) and the Secretary of the Environment and Natural Resources (SEMARNAT). The goal of the
GBAS is to conserve genetic resources of important biological and commercial species of Baja California to help in programs
of conservation or enhance the aquaculture industry in the state.
Up to date, the GBAS contains samples from three important commercial species; red abalone (Haliotis rufescens), Pacific oyster
(Crassostrea gigas), and California flounder (Paralichtys californicus); one endemic species, rainbow trout (Oncorhynchus
mykiss var. nelsoni); and one endanger species, totoaba (Totoaba macdonaldi).
Preliminary studies have been performed for the cryopreservation of sperm of
white shrimp (Litopenaeus vannamei), sea cucumber (Cucumber apostichupus)
and trochophore larvae of red abalone. Other organisms to be considered for
the GBAS are important abalone species such as green, pink, white and black
abalone; clams such as geoduck and pismo clams; different species of sea
urchin, and a variety of marine and fresh water fish.
Sample collection has focus on sperm, embryos or eggs (Fig. 1). However, it
has been considered to store other type of tissues. The acquisition of the samples
depends on organisms’ availability or spawning season. Besides storing samples,
the GBAS achieve different studies to optimize de cryopreservation of endemic
species and carry out techniques to verify sample quality such as cytoplasm membrane
and DNA integrity, changes on proteins (MAPK) activity, and pathogen
transfer, among others (Fig. 2).
Information technologies have been an important issue for the bank. A specific
information management system was developed and called GBAS Information
Management System (GBAS-IMS ℃ ). The GBAS-IMS ℃ provides support for the
operation of the GBAS in daily bases.
The GBAS is a pilot project looking forward to help in the creation of the
National GBAS in the future.
2 1

2 2
CELLULAR AND MOLECULAR IMMUNOMODULATION IN RAINBOW TROUT INDUCED
BY THE DIETARY SUPPLEMENTATION OF Saccharomyces cerevisiae IAM 14383T
A. Panigrahi, V. Kiron, S. Satoh, I. Hirono, T. Kobayashi, G. Yoshizaki, H. Sugita and T. Aoki
Kakdwip Research Centre (Central Institute of Brackish water Aquaculture)
Kakdwip, 24 Praganas (South)
West Bengal, India-743 347
apanigrahi2k@yahoo.com
The present study elucidates the immunomodulation and immune gene expressions in rainbow trout Oncorhynchus mykiss
induced by dietary supplementation of Saccharomyces cerevisiae IAM 14383T. The experimental fish were fed with prebiotic
based diet (basal) incorporated with 10 7 CFU/g of this yeast while the control group feeding basal diet only. The experiment
was conducted for 9 weeks in triplicate tanks in flow through system with 30 animals each.
Realtime PCR assay was performed for looking in to the expression pattern of TNF-α, TGF-β, Interferon (INF), Lysozyme
(Ly) and Immunoglobulin (Ig) genes in tissues like head kidney, spleen, intestine and gill. Further the humoral (serum alternate
complement activity, plasma immunoglobulin) and cellular (Phagocytic activity, super oxide anion of the head kidney leucocytes,
lymphocyte proliferation) systemic immune parameters were evaluated through flow cytometry and other conventional
methods.
The cellular immune response like phagocytic activity and superoxide anion of the head kidney leucocytes is significantly
higher in the treatment group, so also the NK cells increased significantly (p< 0.05). Similiarly the humoral parameters marked
by significant increase (p< 0.05) in the serum alternate complement activity and plasma immunoglobulin levels. The TNF-α
is upregulated maximum in intestine followed by in the spleen and gill whereas the TGF-β mostly unchanged in all tissues but
significantly increased in gill. There is many fold increase in the interferon in the intestine followed by the spleen, gill. The Ig
gene is expressed many fold increase in gill followed by intestine and head kidney. In case of the Ly gene there is many fold upregulation
in the intestine. This study elucidates the immune gene expression pattern in different tissues besides the associated
elicit in humoral and cellular systemic immune response establishing the yeast induced immunomodulation and the possible
mechanism involved in these lower vertebrates.

ORGANIC SHRIMP PRODUCTION: AN ATTEMPT TO BOAST PRODUCTION WITH
SUSTAINABLE APPROACH
A. Panigrahi*, J. Syama Dayal, S. A. Ali, T.K. Ghoshal, P. Ravichandran, K. Ambasankar and A.G. Ponniah
Kakdwip Research Centre (Central Institute of Brackish water Aquaculture)
Kakdwip, 24 Praganas (South)
West Bengal, India-743 347
apanigrahi2k@yahoo.com
High consumer demand for organic aqua-products is leading to many countries taking up organic shrimp farming. In South
East Asian countries including India, in some areas low input traditional systems are being followed, and these can qualify for
organic status. However the farming practices have to meet few more organic farming standards and be part of a certification
system before they can reach global consumers. The present study has looked at to the development of one of the important
components of organic shrimp farming standards, namely the feed. An experimental trial on low input farming of Penaeus monodon
following organic principles was carried out at 6.5 pc/ sq m stocking. In the control feed, the ratio of protein from different
sources (fish meal: other marine protein sources: plant protein sources) was 38: 35: 27 in control feed and 23: 24: 53 in the
low fish meal feed. The total fishmeal percentage in the organic feed was maintained at 15 %. The pond management practice
was based on an earlier trial in which an FCR of 0.95 was achieved by promoting natural productivity. Along with productivity
estimates total protein and amino acid profile were determined to determine if the low protein diet had any negative impact on
metabolic and nutritional needs.
Organic farming resulted in substantial increase of 17 % in production and an increase of 16% in size at harvest (33.28g in organic
compared to 28.64 g in conventional). Average production of 1308 Kg/ha was observed in organic treatments with lower
fish meal feed in comparison to 1116 kg/ha under conventional farming (p

2 4
FEASIBILITY OF FARMING TILAPIA IN THE COOLER ENVIRONMENTS AND THE
EFFECTS OF NUTRACEUTICALS IN IMPROVING THEIR GROWTH
Ashley Parish, Jennifer Taylor and Ahmed Mustafa
Department of Biology
Indiana University Purdue University Fort Wayne
Fort Wayne, IN 46805 USA
Experiments were conducted to investigate the feasibility of farming tilapia in the cooler environments and the effects of
nutraceuticals in improving fish growth. Fish were divided and held in two different environments: warm water (26 ± 2 o C;
80 ± 5 o F) and cool water (15 ± 2 o C; 60 ± 5 o F). Within each environment, half of the fish were provided with nutraceutical
supplemented feed (either vitamin C, zinc, or combined vitamin C and zinc) and the other half of the fish were provided with
regular, commercial feed. Fish held in warm water (control) had better growth, physiological state and immune response than
fish held in cool water, although fish in cool water did not show any differences in condition factor. Fish receiving vitamin C
supplemented feed (700 mg/kg) or zinc supplemented feed (30 mg/kg) or combination of vitamin C and zinc supplemented
feed (700 mg/kg vitamin C and 30 mg/kg zinc) were found to have improved growth (length and weight; p

EFFECT OF STOCKING DENSITY AND FEED RATE ON SURVIVAL AND GROWTH OF
Litopenaeus Vannamei IN AN OUTDOOR FLOW-THROUGH TANK SYSTEM
Susmita Patnaik, Addison L. Lawrence, Brandon Klim and Frank L. Castille
Texas Agrilife Research Mariculture Laboratory at Port Aransas
Texas A & M System
1300 Port Street
Port Aransas, TX 78373 USA
spatnaik3@yahoo.com
Feed management is one of the important components that affects the growth and survival of shrimp. Proper feed rate together
with feed management can be instrumental behind low feed conversion and high growth, survival, and total production. A ten
week growth trial was conducted to evaluate the effect of stocking density and feed rate on growth and survival of Pacific White
Shrimp Litopenaeus vannamei. The study is being conducted in sixty outdoor tanks (and bottom area= 4.374 m 2 ) in ambient salinity
and water temperature. The incoming water was filtered and polished using sand, and diamond filter system before being
used in the tanks. The experiment was conducted with 50 cm of water in all tanks. Tanks were stocked with Litopenaeus vannamei
(5.25 g initial size) which were raised from PLs obtained from Harlingen shrimp farms Ltd., Harlingen, TX. Each tank was
equipped with four airstones (1”x”1x1”) for aeration. The experiment was designed as a three-by-three factorial with stocking
density at three levels (25, 50, and 75 shrimp/m 2 equivalent to 109, 219, and 329 shrimp/m 3 ) and feed rate at three levels (1.0,
1.3, and 1.6 g/shrimp/wk). Shrimp are being fed a prebiotic supplemented (short chain fructooligosaccharide, scFOS), 39% CP
feed three times daily by hand (Zeigler Brothers Inc., Gardners, Pennsylvania). To substantiate the effect of scFOS, a control
treatment was also included where shrimp are offered a feed without any FOS supplementation. Each treatment combination
was replicated six times. To maintain high water quality, the experimental tanks were operated with 33%/d water exchange.
Temperature, dissolved oxygen and salinity are monitored daily. Ammonia, nitrite, nitrate, pH and turbidity were monitored on
a weekly basis. At the study termination, shrimp will be harvested and weighed tank wise separately. Water quality, and shrimp
harvest data including survival, growth, biomass and FCR would be analyzed using a Two-way ANOVA for significance in
difference. The paper will discuss the research findings of the experiment.
2

2
EFFECT OF SOYBEAN MEAL AND SOY PROTEIN CONCENTRATE ON GROWTH
PERFORMANCE OF JUVENILE FLORIDA POMPANO Trachinotus carolinus
Biswamitra Patro* and Robert C. Reigh
Aquaculture Research Station
Louisiana State University Agricultural Center
2410 Ben Hur Road
Baton Rouge, Louisiana 70820-6103 USA
bpatro1@lsu.edu
Florida pompano (Trachinotus carolinus), a marine carnivore, is one of the most expensive food fish in the United States and
is gaining commercial aquaculture importance. Currently, fishmeal is being replaced in fish diets with less expensive plant
proteins such as soybean meal and its derivatives to reduce the feed cost and subsequent production cost. A 12-wk feeding trial
was conducted to determine the effect of different levels of soybean meal (SM) and soy protein concentrate (SC) on the growth
performance of juvenile Florida pompano. A control diet was formulated with dehulled solvent-extracted soybean meal, lowantigen
soy protein concentrate, wheat flour, corn gluten meal, and fishmeal, to contain 45% crude protein and 12% crude fat.
Another five experimental diets (SC36/SM35, SC39/SM30, SC42/SM25, SC46/SM20, and SC58/SM0) were formulated with
the same ingredients except fishmeal to match the control diet’s protein and fat content. Fifteen pompano juveniles (mean body
weight of 3.7 g) were stocked in each of 24, 189-L rectangular tanks in two recirculation systems maintained at about 16 ppt
salinity. Each of the six diets was randomly assigned to two tanks in each system. The fish were fed to apparent satiation everyday
for 12 weeks and weighed at the end of 2, 4, 8, and 12 weeks. There was no difference in the growth performance among
fish fed the non-fishmeal diets and for each weighing period, all fish fed non-fishmeal diets had significantly (P < 0.05) lower
weight gain and specific growth rate than fish fed the control diet. To determine if the non-fishmeal diets had any palatability
issue and consequent lower growth rate, another feeding trial was conducted with selected experimental diets from the first trial
which were reformulated to contain some fishmeal. The data from both the feeding trials will be presented and discussed.

EVALUATION OF THE EFFECTS OF TEMPERATURE AND BODY MASS ON
RESPIRATION RATES OF PADDLEFISH Polyodon spathula USING RESPIROMETRY AND
DISSOLVED BLOOD GASES
Joshua T. Patterson*, Steven D. Mims, Richard J. Onders and Russell A. Wright
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
joshua.patterson@kysu.edu
Bioenergetics modeling is a valuable tool for assessing the trophic level dynamics in fish communities. These models are used
to estimate predator-prey relationships and food consumption. A significant component of any bioenergetics model is respiration.
An extensive production method for paddlefish known as reservoir ranching is currently in research and development
stages. The paddlefish is an emerging aquaculture species valued for both its firm, boneless, white meat and dark roe which
is processed into caviar. Development of a bioenergetics model for paddlefish is seen as a crucial step in expansion of reservoir
ranching. In the present study, paddlefish respiration was evaluated over the range of water temperatures found annually
in large reservoirs of the central United States, and over the range of weight classes attained by paddlefish during a growth
cycle.
Static respirometry was used to allow unrestrained paddlefish to swim at a constant speed. Dissolved oxygen (DO) concentrations
were monitored and recorded over time with a Strathkelvin Instrument meter mounted in two static respirometers of 500
L and 2300 L. Three water temperatures were used in the study: 10, 20, and 30°C. Paddlefish of three size classes were tested:
0-1.5 kg to correspond with stocker-size fish initially placed in reservoirs, >1.5 to 4.5 kg to correspond with fish in the 3 to 4
year age range, and >4.5 kg to correspond with harvest size fish. The smallest size class was tested in the 500-L respirometer
while the other size classes were tested in the 2300-L respirometer. Each size class was tested at each temperature with 8 to
10 replicates per trial group. Also, five fish from each size/temperature pairing were held in water with DO at saturation while
blood samples were drawn from the caudal vein. Subsequently, blood was drawn from at least five fish in each group after
spending time in the respirometer. Blood pH, PCO 2 , and PO 2 were compared between groups held at saturation and those from
the respirometer. Some results are presented below and will be discussed.
10° C Avg. 02
Consumption
(mg/h/kg)
Avg.
Temp
(° C)
Avg.
Weight
(kg)
20°
C
>4.5 kg 59.23 11.38 11.04 >4.5
kg
>1.5-4.5 80.08 10.38 3.80 >1.5kg
4.5
kg

2
THE FDA’S ANIMAL FEED SAFETY SYSTEM: ENSURING THE SAFETY OF ANIMAL
FEED (INCLUDING AQUACULTURE) AND PET FOOD
Frances M. Pell, M.S.,
U.S. Food and Drug Administration,
Center for Veterinary Medicine
Division of Compliance
Rockville, MD 20855 USA
frances.pell@fda.hhs.gov
The Food and Drug Administration (FDA) began modernizing its animal feed safety program in 2003. The new program, the
Animal Feed Safety System (AFSS), is being designed to be comprehensive, preventive, and risk-based so that the FDA and
collaborating states can ensure the safety of feed intended for food animals (including aquacultured) and pets, as well as the
safety of human food derived from food-producing animals. The AFSS has six components: “Ingredients and the Approval
Process,” “Limits for Animal Feed Contaminants,” “Process Control for the Production of Feed Ingredients and Mixed Feed,”
“Reporting of Unsafe Feed,” “Regulatory Oversight,” and “Education and Outreach.” These are all presented in the AFSS
“Framework Document,” which is available on the Web site of FDA’s Center for Veterinary Medicine. The Framework also
identifies gaps for each component as well as the manner in which the FDA intends to address each gap. To incorporate the
concepts of risk-assessment in AFSS, the AFSS Team developing the program has drafted a risk-assessment tool. The Team
is continuing to collect data that can be used for the model and is working to validate the model. AFSS has also been revised
to include provisions of the FDA Amendments Act of 2007 concerning the safety of pet food and feed ingredients. The AFSS
initiative fits well into FDA’s overarching Food Protection Plan, which was designed to integrate all federal, state, and local
food safety and food defense (counterterrorism) programs in the United States. AFSS and the Food Protection Plan have
many “cross-cutting” principles. Detailed information on the AFSS project is available at the following link: http://www.fda.
gov/cvm/AFSS.htm

EVALUATE THE OF PLANKTONIC AND BENTHIC COMMUNITIES OF MICROALGAE IN
THE SHRIMP POND (Litopenaeus vannamei)
João Batista Pereira Neto, Danielli Matias de Macêdo Dantas, Alfredo Olivera Gálvez and Luis Otavio Brito
Pernambuco Federal Rural University
Fisheries and Aquaculture Department
Dois Irmãos, 52171-900
Recife, Pernambuco, Brazil
engpescalo@hotmail.com
The present study aimed to evaluate the of planktonic and benthic communities of microalgae, and establish relationships with
the hydrological variables, management practices and the production data of the shrimp ponds.
During of period the 2005 September the 2006 september, three Litopenaeus vannamei ponds and the affluent channel were
analyzed by taking phytoplankton and benthos samples twice a week.
In the ponds were recorded information’s about the hydrological variables, climatic, culture management and the production
data.
The Cyanobacteria dominated the phytoplankton in all cycles, reaching the relative abundance of 80%, being the Pseudoanabaena
genus the main responsible for this domination. In the benthos, the Diatoms dominated with abundance over 85%.
The phytoplankton reached higher densities in the ponds, whereas the phytobenthos was dominant in the affluent channel. A
positive relation between the benthos diatoms and the shrimps´ growth was observed.
2

2 0
EVALUATION OF WINTER FEEDING ON EXPERIMENTAL CHANNEL CATFISH
GROWOUT POND PLANKTON
Peter Perschbacher,* Gerald Ludwig and Adam Nanninga
Aquaculture/Fisheries Department
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601 USA
pperschbacher@uaex.edu
Ten, 0.1-ha ponds at the UAPB Aquaculture Station were sampled weekly from Dec. 7-Feb. 22 (n=90) to test the effect of winter
feeding on phytoplankton and zooplankton. Five of the ponds were randomly assigned to each of two treatments: no feeding
and feeding based on recommended rates. Channel catfish sizes and numbers approximated a typical winter carryover scenario.
Chl. a (biomass) and variability was low and not significantly different (Table 1). Average combined algal natural units were
more than twice as high in fed ponds (P=0.02). Composition between treatments was not significantly different. Diatoms were
the most abundant group. Zooplankton did not differ. Off-flavor cyanobacteria were two species of Anabaena and Pseudanabaena
sp. These off-flavor algae were found on 7 of the 9 sampling dates in fed ponds and 9 of 9 in unfed ponds and occurrence
was significantly higher in unfed ponds (P=0.08) by 70%.
Thus, algae in fed ponds were smaller and off-flavor algae less common than in unfed ponds. Winter feeding of channel catfish
may benefit algal composition.
Table 1. Effect of winter feeding on plankton in experimental catfish production ponds (significance indicated by *).
Parameter fed unfed
Chl. a (ug/l) 82.5 + 56.2 81.2 + 53.5
Pheophytin a (ug/l) 35.9 40.9
Total algae (#/ml) 11,506 4,851*
Algal composition (%)
Diatom 63.5 57.7
Green 12.9 18.1
Euglenoid 15.4 13.8
Cyanobacteria 10.4 7.8
Off-flavor algae occurrence
(% of samples) 22.2 37.8*

RISK EVALUATION TO AQUACULTURE PONDS FROM COMMON AERIALLY-APPLIED
HERBICIDES IN THE MISSISSIPPI ALLUVIAL PLAIN (DELTA)
Peter W. Perschbacher,* Gerald M. Ludwig and Regina Edziyie
Department Aquaculture/Fisheries
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601
pperschbacher@uaex.edu
Common row crop herbicides were tested for possible adverse effects from drift to aquaculture production pond plankton and
water quality in triplicate 500-L outdoor, pool mesocosms.
Production pond systems greater than 7 ha. Forty aerially-applied herbicides were tested, including 11 soybean, 13 rice, 10
cotton, 5 corn and 1 winter wheat at low (1% of field rates) and high (10%) drift estimates for ponds 7ha and greater (ie channel
catfish). Diuron (cotton) affected phytoplankton and oxygen production in relation to dose. Morning oxygen levels fell to
critical levels of 3 ppm and below after one week in the high drift treatment. Zooplankton were also reduced relative to control
levels at both drift levels. Atrazine (corn) reduced zooplankton numbers. Carfentrazone (soybean) also reduced zooplankton,
but slightly
Positive impacts. Diuron, propanil and atrazine reduced larger cyanobacteria (bluegreen algae), pH and thus unionized ammonia
(UIA). Propanil and atrazine stimulated green algae and diatoms, as well as chlorophyll a, after initial reductions.
Other species pond systems. Greater impact was noted from propanil on systems with midlevel algal levels (50-200 ug/l as in
hybrid striped bass and catfish ponds) than in low and high chlorophyll a levels (below 20 and above 300 ug/l as in freshwater
shrimp and goldfish ponds, respectively). Drift levels were estimated up to 20% due to pond sizes typically less than 7 ha.
Algal scum conditions. Scums were more susceptible to propanil than populations mixed in the water column and impacts
approximated direct application levels (ie. overspray). Bloom instability and greater contact were assumed responsible.
Fry and fingerling pond systems. Smaller fry ponds were found to have potential drift levels of up to 3 times larger production
ponds. Greater effects were found accordingly, however chlorophyll a was low and reduced the impacts of atrazine and propanil.
Ph and UIA were also reduced. Thus, little impact was expected on fry production. Diuron however reduced DO below
3 ppm for on the second and third days. Recovery in DO was noted after two weeks. Thus, additional aeration may be needed
following diuron drift. Cladocerans were also reduced and may affect production.
2 1

2 2
ULTRASTRUCTURAL STUDY OF THE NUCLEOLAR CYCLE DURING
SPERMATOGENESIS OF Tilapia rendalli (TELEOSTEI, CICHLIDAE)
Rita L. Peruquetti * , Sebastião R. Taboga and Maria Tercília V. Azeredo-Oliveira
São Paulo State University
Departmento of Biology
São José do Rio Preto, SP 15054-000
ritaperuquetti@yahoo.com.br
Chromatoid body (CB) is a cytoplasmic structure of germ cells that has probable role in the RNA and protein reserve in the
different stages of spermiogenesis. Some authors believe that CB is formed by extrusion of nucleolar material from the nucleus
to the cytoplasm. This nucleolar cycle has been studied during spermatogenesis of some animal groups. The aim of the present
study is to follow the nucleolar cycle in fish. The used specie was the Tilapia rendalli, specie brought from Congo (Africa) to
Brazil, in 1953 and that, currently, it meets spread in all the dams of Brazil widely. The CB formation is an important aspect of
the animal reproduction. We need to improve our knowledge on this process and to know how it happens in T. rendalli, specie
that has a great economic importance and is widely cultivated in captivity in Brazil. The testes of each animal were removed,
sliced into small pieces and samples of the seminiferous tubules were minced and fixed by immersion with 3% glutaraldehyde
plus 0.25% tannic acid solution in Millonig’s buffer (pH 7.3) containing 0.54% glucose for 24 hours. After washing with the
same buffer, samples were post-fixed with 1% osmium tetroxide for 1hour, washed in Millonig’s buffer, dehydrated in a graded
acetone series, and embedded in Araldite resin. Ultrathin silver sections were cut using a diamond knife and stained with 2%
alcoholic uranyl acetate for 30 minutes followed by 2% lead citrate in sodium hydroxide for 10 minutes. Samples were evaluated
using a Leo-Zeiss 906 (Cambridge, UK) transmission electron microscope (TEM). Cytoplasmic “nuages” was originated
in spermatogonia when the nucleoli is organized yet (Figure 1A). CB had it’s major volume in the cytoplasm of primary spermatocytes,
after the nucleolus disorganization. CB was observed in association with mitochondrial aggregates in this type of
cell (Figure 1B). Later spermatids presented CB in migration to axonema region (Figure 1C) and the nucleolus was reorganized
with a small volume than the spermatogonia nucleolus. In the mature spermatozoon no CB material was found (Figure 1D). In
conclusion, data demonstrated that there is disintegration of the nucleolus and a fraction of this nucleolar material, probably,
migrates to the cytoplasm, where a specific structure is formed, called chromatoid body. This cytoplasmic structure participates
in some parts of spermiogenesis process: tail and middle piece formation of the spermatozoon, cellular communication between
spermatids.
Figure 1: A – Spermatogonia. B – Primary
Spermatocyte. C – Later Spermatid. D –
Spermatozoon. (n: nucleus; nu: nucleolus; *
and cb: chromatoid body material; mi:
mitochondria; f: flagellum. Bars = 10 µm.

EVALUATION OF CHANNEL CATFISH (Ictalurus punctatus) PITUITARY FOR THE
INDUCED SPAWNING OF GRASS CARP (Ctenopharyngodon idella)
Mark A. Peterman*, Ronald P. Phelps and Jason S. Broach
E.W. Shell Fisheries Research Center
Department of Fisheries and Allied Aquaculture
Auburn University, Auburn, AL 36849 USA
Peterm4@auburn.edu
The US catfish industry processed almost 500 million pounds of fish in 2007 and in doing so also generated a considerable
volume of fish wastes, currently of little economic value. One item that might be recovered as a valuable by-product is catfish
pituitary. Pituitaries of several fish species, mainly common carp, are used for induced spawning of fish. Common carp
pituitary is widely used and effective in a number of fish species. Channel catfish (Ictalurus punctatus) pituitary has been used
effectively in the induced spawning of channel catfish. However, it has not been evaluated in many other species. In this study
the efficacy of channel catfish and common carp pituitary for induced spawning of grass carp (Ctenopharyngodon idella) were
compared.
Pituitary glands were collected from channel catfish carcasses at a commercial catfish processing facility and processed into a
dry powder. Female grass carp (2.4 ± 0.74 Kg) were injected with 4 mg/kg of catfish or carp pituitary, giving 10% as a preparatory
dose followed with a resolving dose (90%) twelve hours later. Males (1.9 ± 0.55 Kg) were given 2 mg/Kg of either pituitary
at the time of the resolving dose for the female. Water temperature ranged from 22.9°C to 23.7°C.
Females ovulated approximately 10 h after the resolving dose with no difference in time of ovulation related to type of pituitary
given. On average, 84 % of the females ovulated with no difference between the two types of pituitary used. The type of
hormone used did not influence any of the other reproductive parameters measured. Females released an average of 83,831 ±
25,442 eggs/Kg, and the average number of eggs/g was 679.6 ± 138.8 eggs/g. Egg viability averaged 85.6 ± 11.96%. Males
performed similarly when given catfish or common carp pituitary, giving a mean volume of 2.96 ± 1.2 ml of semen/Kg body
weight, and sperm motility averaged 79.4 ± 26.6 seconds.
These results suggest that catfish pituitary is a viable option for use in the induced spawning of grass carp. In addition, these
results show promise for the future investigation of channel catfish pituitary for use in the induced spawning of other fish species.
2 3

2 4
COMPARISON OF GROWTH, BODY COMPOSITOIN, AND STRESS RESPONSE OF THREE
SELECT LINES AND INDUSTRY CHANNEL CATFISH
Brian. C. Peterson*, Brian G. Bosworth and Brian C. Small
USDA-ARS Catfish Genetics Research Unit
Thad Cochran National Warmwater Aquaculture Center
PO Box 38
Stoneville, MS 38776 USA
bpeterson@ars.usda.gov
Improvement in growth and resistance to stress has been identified as traits of interest for genetic improvement. Channel
catfish selected solely for fast growth (fast growing) were compared to USDA103, USDA403, and industry pool groups
of channel catfish for growth, body composition, and stress response. All fish were fed daily for 8 wks followed by an
acute 10-minute dewatering stress. By wk 4, feed intake and weight gain were significantly different for each genetic group
(P < 0.0001). Overall, feed consumption and weight gain were highest in the fast growing line of fish (P < 0.01). Feed conversion
ratio was lower in the fast growing fish compared to the USDA103 line (P < 0.05). Whole body proximate composition
and nitrogen retention were similar among lines of fish. Fast growing, USDA103, and USDA403 lines had higher condition
factors and protein efficiency ratios compared to industry pool fish (P < 0.05). Plasma cortisol levels were similar among
genetic groups after an acute stress. These results demonstrate improved growth of the USDA103 line of catfish and lines
derived from it over the industry pool.

GREENHOUSE GAS EMISSIONS ASSOCIATED WITH DIRECT ENERGY INPUTS FOR A
WARMWATER LOW-SALINITY RECIRCULATING AQUACULTURE SYSTEMS
Timothy J. Pfeiffer and Paul Wills
USDA Agricultural Research Service
Fort Pierce, FL. 34946 USA
timothy.pfeiffer@ars.usda.gov
Greenhouse gases (GHGs) are gases that trap heat in the atmosphere. These gases include carbon dioxide (CO 2 ), methane
(CH 3 ), nitrous oxide (N 2 O), and fluorinated gases. Some of these gases occur naturally and some are created by human activities
which can increase their concentrations. The most common method of GHG concentration increase is through the combustion
of fossil fuel for electricity. CO 2 is the predominant GHG in the atmosphere and often the focus of greenhouse gas
emission discussion although N 2 O gas emissions have 300 times more heat trapping capabilities over a 100 year time period.
In the state of Florida the governor has issued executive orders for the reduction of greenhouse gases emission levels. By 2050
electric utilities will need to reduce GHG emission to 80 percent of 1990 levels. State agencies and departments are targeted
for a 40% reduction from current emission levels by 2025. Given the GHG emission reduction pressure from state and federal
administrations a GHG assessment was undertaken of the current recirculating aquaculture systems in operation at the USDA
Agricultural Research Service Sustainable Marine Aquaculture project on the Harbor Branch Oceanographic Institute campus
in Fort Pierce, FL.
The low-salinity recirculating aquaculture systems (RAS) that were evaluated for GHG emission range in system volume from
10 m 3 to 45 m 3 . The rearing options in these systems include fingerling to juvenile production and juvenile to grow-out production.
In the two juvenile culture RAS units tank diameter is 1.52 m and the number of tanks in each system were nine and
ten respectively. System components include UV sterilizers, centrifugal and propeller pumps, swirl separators, polygeyser and
moving bead biofilters, and foam fractionators. In the grow-out production RAS units tank diameter is 3.05 m and the number
of tanks in each of the eight RAS units is four. System components include drum filters, propeller-wash bead filters, moving bed
biofilters, UV sterilizers, centrifugal and propeller pumps, degas and oxygenation towers, and oxygenation cones. The systems
have been utilized for red drum, florida pompano, hybrid striped bass, and cobia grow-out studies.
The direct energy inputs of the RAS units utilized to determine the GHG emission levels in terms of metric tons of carbon
dioxide produced include the energy to pump, treat, and transport the water in the systems. The intent is to be able to compare
the different system treatment designs and operation in some form of sustainability perspective and develop a GHS Reduction
scorecard for energy and water savings of the various RAS unit designs.
2

2
A PRELIMINARY ASSESSMENT OF PROTEIN SYNTHESIS USING DEUTERATED WATER:
A NOVEL APPROACH TO CHARACTERIZE GROWTH OF CHANNEL CATFISH Ictalurus
punctatus
Camilo Pohlenz*, Heath Gasier, Michael Wiggs, Alejandro Buentello, Stephen Previs, James Fluckey and Delbert
Gatlin III
Texas A&M University System
College Station, TX 77840-2258 USA
cpohlenz@tamu.edu
A thorough understanding of protein synthesis (PS) in the skeletal muscle of fish is needed to improve the efficiency of aquaculture
production. In fish, muscle accounts for over 50% of total mass of most species, and is composed mainly of protein.
Traditionally, PS in fish has been assessed with a procedure known as the “flooding dose”. However, concerns about the
suitability of this method have surfaced as a result of sharp variations obtained for whole–body PS estimates. These variations
likely result from the brevity of treatment and nutritional state of the animal at the time of measurement. Deuterium oxide
( 2 H 2 O) is a versatile stable isotope tracer that makes possible the study of bio–molecule dynamics with minimal disturbance
to the living organism, and permits long–term investigations which yield integrative measures of PS. Therefore, the specific
objective of this study was to evaluate fractional synthetic rates (FSR) of muscle PS in juvenile channel catfish. To this end,
the incorporation of 2 H from 2 H 2 O administered in–vivo into L–alanine was used to compute FSR.
A preliminary experiment to evaluate the equilibration of plasma H 2 O with 2 H 2 O used 24 fish (~65 ± 1.2 g each) in a 30–L
aquarium. 2 H 2 O was added to the water at a 4.0% concentration. Samples of six randomly selected fish were taken at 0, 4,
8 and 24 h for extraction of blood from the caudal vein and muscle from the left epaxial myomere. Separated plasma and
dissected muscle samples were rapidly stored frozen at –80 C, until further analysis. In a second experiment, the effects of
dietary arginine (Arg) on muscle FSR were evaluated. After a 6–week feeding period with two different levels of L–Arg•HCl
(0.5 and 4.0% of diet), 2 H 2 O was added to the aquarium water and six fish/diet were sampled at 24 h, as described above. Gas
chromatography/mass spectrometry was used to assess the 2 H–labeling of plasma water and muscle protein.
Table 1 indicates that 2 H equilibrated in catfish plasma within 1 h, and remained stable for ~ 24 h. There was a continuous
increase in the 2 H–labeling of muscle protein. FSR was only calculated from the 24 h samples. No significant differences
(P > 0.05) were observed between FSR of fish fed different levels of Arg. Taken together these results indicate that the assessment
of PS over an extended period of time is easily attained in catfish using deuterated water. Some advantages of using of
2 H2 O to evaluate PS are that it is a non–invasive and relatively inexpensive method, accounts for the influence of feeding; and,
the ease of administration makes it a practical assessment option.

MULTIPLE COMPARISONS OF MEANS VS. REGRESSION ANALYSIS IN AQUACULTURE
RESEARCH
Steeve Pomerleau
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
Mail Slot 4912
1200 North University Drive
Pine Bluff, AR 71601 USA
spomerleau@uaex.edu
Comparing treatment means by testing for statistical significant differences is probably the most common approach used in
aquaculture research. However, in several common types of aquaculture experiments such as in fertilization experiments or
stocking density experiments, where treatments correspond to several levels of a quantitative or continuous variable, comparing
treatment means for significant differences is not an appropriate and efficient way of analyzing the data and of obtaining
meaningful information. In those types of experiments, regression analysis should be used to estimate and identify trends or
relationship instead of simply comparing means to find which treatment gave the best results. Unfortunately, in the literature,
multiple comparisons of treatment means are too often unwarranted and the regression analysis is too often ignored where it
would have been the most appropriate approach for the factors being studied. Comparing treatment means is not necessarily
wrong; it is just that more valuable information could sometime be attained with the same amount of resources if a regression
analysis and a slightly different experimental design were used.
This problem has been raised several times in the literature. Chew (1976) wrote: “Duncan’s multiple range test (MRT) is very
often inappropriately used to compare treatments that are factorial in nature or that correspond to several levels of a quantitative
or continuous variable.” Lowry (1992) wrote: “Pairwise, multiple comparisons are appropriate only for comparing unstructured,
qualitative treatments.” Dawkins (1983) wrote: “How do we persuade biologists of the futility of significance testing
between mean responses (ordinate value of y), at series of dose-rates (abscissa values of x), along response curves?” Knud-
Hansen (1997) wrote: “Although the statistical community is quite clear on this point, the aquaculture community (among
other scientific disciplines) has yet to appreciate the inappropriateness and gross inefficiency of multiple range test usage in
analyzing structured experiments.”
2

2
ELECTRIC RATES IN THE CATFISH INDUSTRY
Steeve Pomerleau
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
Mail Slot 4912
1200 North University Drive
Pine Bluff, AR 71601 USA
spomerleau@uaex.edu
Electric rate schedules from several electricity providers in Alabama, Arkansas, and Mississippi were studied to quantify the
differences in electric rates paid by catfish farmers. Rate schedules can be quite complex and composed of numerous types of
charges. All those charges need to be taken into account when comparing rate schedules. There are six major types of charges:
1) energy charges in $/kWh, 2) demand charges in $/kW, 3) customer charges in $/month, 4) minimum charges, 5) taxes, and
6) rider schedules. Rider schedules are designed to adjust another rate schedule. They are often overlooked but can constitute a
major part of a customer electric bill.
As of June 2008, only three of the nine studied electricity providers offered a time-of-use schedule (with different on-peak and
off-peak hour rates) to catfish farmers: Alabama Power Co., 4-County Electric, and Entergy Mississippi Inc. The rate schedule
of Alabama Power Co. resulted in the lowest total annual electricity costs (29 to 46% lower than other studied rate schedules).
The off-peak energy rate of Alabama Power Co. was 2.3893 ¢/kWh, but when all charges were taken into account the electricity
cost was estimated at 6.5 ¢/kWh as opposed to an average of 10.8 ¢/kWh for the eight other studied schedules. However,
results may vary. Electric rates change frequently. Fuel cost adjustment riders and other rider schedules may change monthly
or quarterly. Besides, the results do not reflect some credits and/or rebates issued by some Mississippi electric cooperatives in
recent years. These credits were funded by credits from the South Mississippi Electric Power Association (SMEPA).
Table 1. Summary of studied 2008 electric rates applicable to catfish farmers and estimated annual electric costs for a block of
four ponds with one well and eight 10-hp paddlewheel aerators.

PUTTING TOGETHER A BUSINESS PLAN
Steeve Pomerleau
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
Mail Slot 4912
1200 North University Drive
Pine Bluff, AR 71601 USA
spomerleau@uaex.edu
In considering an aquaculture business venture, there are a number of important decisions to be made. This presentation is
intended to help potential producers by describing the components of a good business plan necessary to evaluate feasibility,
estimate operating capital, and secure adequate financing. A good business plan will include: 1) a description of the proposed
site and production system, 2) a marketing plan, and 3) a financial analysis. The financial analysis should include: a) enterprise
budget, b) estimate of required financing, c) farm appraisal, d) balance sheet, e) income statement, f) cash flow budget, and g)
personal financial statements.
Aquaculture is capital intensive and will often require million of dollars. A successful application for a loan for an aquaculture
operation must be well documented. A good business plan will help secure adequate financing especially when lenders are not
familiar with aquaculture. Besides, adequate planning for the business may help avoid mistakes and minimize risks associated
with the market, production and financing.
The marketing plan is often the most overlooked component of a business plan. Many growers focus on the technical aspects of
fish production and do not spend time considering market opportunities. Decisions on species, harvest size, and volume should
be based on a market analysis. The most successful aquaculture businesses are those that are market-oriented, have diverse
markets, and are committed to their customers. Profits are made by selling fish, not producing them.
The financial analysis will help you determine if the business will generate enough cash revenue when expenses are due and
payable. A feasible plan will indicate potential profits, but will the level of profit make your efforts worthwhile? Fish farming
is a great way of life but requires a great deal of preparation and planning. A business plan is the future of the business on paper
and a way to test dreams against expected challenges and opportunities.
2

2 0
WHAT IS THE EFFECT OF DIFFERENT INDISPENSABLE AMINO ACID IMBALANCED
DIETS ON TROPICAL FISH PACU Piaractus mesopotamicus ?
Rodrigo Takata, Maria Célia Portella, Thiago M. de Freitas, Karolina Kwasek and Konrad Dabrowski
Sao Paulo State University - Aquaculture Center
Via Prof. Paulo Donato Castellane
14884-900 Jaboticabal, SP, Brazil
portella@caunesp.unesp.br
Under practical farming conditions, feed formulations that include plant proteins may result in disproportionate amounts and
deficiencies of indispensable (essential) amino acid (IDAA). The pacu, Piaractus mesopotamicus, is a native species extensively
cultured in Brazil and frequently subjected to disproportionate amounts of IDAA in commercial diets. The present study
aimed to investigate how disproportional amounts of IDAA in diets for juvenile pacu affects the growth and survival when
IDAA-imbalanced diets are provided in subsequent meals. The experiment was carried out for 56 days. Four isonitrogeneous
and isolipidic diets were used: casein-gelatin-based diet (protein control), FAA-based diet (with all indispensable amino acids),
(-)Lys diet (devoid of Lys, His, Ile, Phe and Trp and (-)Arg diet (devoid of Arg, Thr, Val, Leu and Met). Amino acids were
supplied in the form of L-free amino acids in FAA, (-) Lys and (-)Arg diets. Fish were fed four times per day at 4-hour intervals
and the feed schedule (treatment) used were: (-)Lys(-)Arg group, fish were fed 4 meals of (-)Lys, (-)Arg, (-)Lys and (-)Arg
diets each day throughout the experiment; (-)Lys(-)Lys group, fish were fed 2 meals of the (-)Lys diet as first two meals in the
morning and then 2 meals of the (-)Arg diet in the afternoon; (-)Lys/(-)Arg group, fish were fed four meals of the (-)Lys diet and
the (-)Arg diet on alternate days; FAA and Protein diets (controls); fish were fed FAA-based balanced and casein-gelatin based
diets, respectively, each day throughout the experiment. Mortality was monitored and moribund fish were removed to avoid
cannibalism. Growth performance was assessed by using mean individual weight. Fish fed the protein diet showed the highest
(P

MORPHOLOGICAL DESCRIPTION OF THE INITIAL DEVELOPMENT OF PIAUÇU
LARVAE Leporinus macrocephalus
Maria C. Portella*, Natalia J. Leitão, Márcia R. F. Machado, Sérgio T. J. Tamassia, Fernanda Q. Silva,
Adriana F. Barros and Rodrigo M. R. Sousa
Aquaculture Center
Sao Paulo State University
Jaboticabal, SP 14.884-900
portella@caunesp.unesp.br
The larval period includes a succession of important changes in the structure and function of most of organic systems. Important
events as yolk absorption and the onset of exogenous feeding occur in this phase. This study describes the initial development
of piauçu larvae Leporinus macrocephalus and was conducted as an activity during the Fish Larviculture discipline, offered by
the Pos-graduation Programs of Animal Science and Aquaculture of the Sao Paulo State University – UNESP, SP, Brazil.
Larvae were sampled from hatching still the 7 th day post hatching. From day four onwards, they were fed on Artemia nauplii.
Fifteen larvae were fixed in buffer formalin 4% solution (pH 7.4) for 24h. The fixed material was used for determination of
yolk-sac volume (mm 3 ) and wet weight (mg). Ten larvae were anaesthetized and used for description of the morphological
changes.
2 1

2 2
EFFECTS OF EARLY WEANING ON SURVIVAL OF Salminus brasiliensis
Maria Célia Portella*, Mônica Giacometti Mai, Rodrigo Takata, Olívia Cristina Camilo Menossi,
Thiago Mendes de Freitas, Natalia de Jesus Leitão and José Roberto Verani
Sao Paulo State University - Aquaculture Center.
Via Prof. Paulo Donato Castellane
14.884-900 Jaboticabal, SP, Brazil
portella@caunesp.unesp.br
The dourado Salminus brasiliensis is a tropical fish species endangered by pollution, overfishing and by dam construction,
which prevent its upstream migratory reproduction journey. It is known that the early development of S. brasiliensis in captivity
is the most critical period for its production, and no rarely mortalities of 90% are verified. The weaning process from live food
to formulated diet is highly recommended for this species but little information is available. The objective of this study was to
evaluate the effects of early weaning of S. brasiliensis using different types of attractive feeds.
The weaning last 5 days beginning at the fourth and ending at the eighth day after the onset of the exogenous feeding. Larvae
were fed four times a day (6 am, 12 am, 6 pm and 12 pm). Dourado larvae were fed on prey larvae of Leporinus obtusidens (P),
Artemia nauplii (A), frozen biomass of Artemia (B), commercial diet (CD), and beef (bovine heart) (H), all of them in gradual
(co-feeding) or abrupt weaning. After have been weaned, larvae were fed on formulated diet for one more week to evaluate the
adaptation of the fish to the commercial diet.
The results concerning the survival are shown in Fig.1. Due to the failure of the gradual weaning protocol, a comparison with
the abrupt weaning was not performed, but some interesting survival rates were still obtained. Despite the significant differences
in weight and length (only observed in abrupt schedule) among the treatments, the analyses of relative condition factor
(Kn = W t /a.L t b ) resulted in no significant differences among them. In the co-feeding treatments it was observed an increase of
the ammonia levels, but not reaching the values considered lethal for dourado juveniles. Even though, in some tanks high mortalities
were observed, indicating that the larvae are more sensitive to ammonia than the juveniles. In the treatment were beef
was offered, total mortality was observed in the abrupt weaning schedule and poor results in the gradual transition schedule
(1.75 % survival) at the end of the experiment. These treatments were excluded from the statistical analysis.
These results can be used as a guide of the type of feed that can be offered in the early stages for the species. In this way, a
commonly used feed as beef showed to be inefficient, and Artemia nauplii in abrupt weaning schedule showed to be a possible
solution in cases of insufficient fish prey larvae in the beginning of the dourado’s development.
Survival after Abrupt
Weaning
100
80
a
60
40
20
0
b
b
C B A P
a D
b
Treatment
s
Survival after Cofeeding
100
50
0
b b
B+C
A+C
D D D
Treatment
c d
s
a
a
P+C
Survival After Subsequent
Week (Abrupt
Schedule)
4
0
2
b b b
0
0
C B A P
D
Treatment
s
Survival After Subsequent
Week (Co-feeding
3 Schedule) a
0
2
0
1
0
0
b b
B+C A+C P+C
D Treatment D
s
D
Fig.1 – Survival rates (%) of S. brasiliensis
larvae.
a

DETERMINING SIZE EFFECTS ON PHOSPHORUS UPTAKE, GROWTH AND
PHOSPHORUS METABOLISM IN RAINBOW TROUT FRY, FINGERLING AND POST
JUVENILE STAGES
Madison S. Powell*, Gordon K. Murdoch, Frederic T. Barrows, Jurij M. Wacyk, Michael T. Casten,
Wendy M. Sealey, David A. J. Stone and Ronald W. Hardy
University of Idaho
Aquaculture Research Institute
Hagerman Fish Culture Experiment Station
3059-F National Fish Hatchery Road
Hagerman, Idaho 83332 USA
Sufficient dietary phosphorus (P) is essential for bone formation and a range of metabolic processes in fish that support optimum
growth. Rainbow trout feeds are typically formulated based on P availability of commonly used ingredients which range
from approximately 40 to 60%, and an available dietary P requirement of 0.66%. However, the P requirement data were determined
for 200 g rainbow trout. This requirement level is used for feeds for smaller rainbow trout because P availability and
requirements of first feeding and small rainbow trout are unknown. However, dietary nutrient requirements of small fish are
often greater than those of large fish. Thus, rainbow trout at different stages of production may require different dietary P levels.
In this study we measured P availability in rainbow trout over their entire production cycle, examined bone mineralization
and also compared genetic expression of key enzymes of P metabolic pathways in rainbow trout in response to varying levels
of dietary P. Three diets were formulated with P availabilities of 0.33, 0.66, and 1.32%. Diets were fed to triplicate tanks of
rainbow trout over three separate phases comprising first feeding to 20 g fish (Phase I), 20 g to 200 g fish (Phase II), and 200 g
to 500 g fish (Phase III). Overall growth was not significantly affected by dietary P availability until the end of the study when
fish reached market size (Phase III). However, skeletal mineralization and gene expression associated with phosphorus availability
differed significantly when rainbow trout were between 0.13 to 20 g suggesting that dietary P requirement for optimum
physiological health of fry may be higher than that of juvenile trout.
2 3

2 4
AQUAFRIN: A NOVEL PHOTO-ACTIVATED ANTIMICROBIAL AGENT FOR
AQUACULTURE
Glenn D. Prestwich, Jerry Bommer and Jason Henderson
Frontier Scientific, Inc.
P.O. Box 31
Logan, UT 84323 USA
An unmet need in aquaculture is a broad-spectrum, easy to use antifungal/antibacterial agent that is (i) safe, (ii) has low vertebrate
toxicity, (iii) does not select for resistance, (iv) is free of hazardous peroxides, radicals, and odors, and (v) does not harm
plants, algae, cyanobacteria, or aquatic invertebrates. Such an agent should be environmentally benign and have a degradation
profile consistent with use in both indoor and outdoor fishery facilities. We now describe results for AquaFrin (Lauryl Methyl
Pyrifrin), a water-soluble and amphiphilic porphyrin that meets these criteria. AquaFrin is a photosensitizer that disrupts
bacterial, protozoal and fungal cell walls and membranes by local production of singlet oxygen, a potent oxidant that diffuses
less than 0.1 mm before. Structure activity studies demonstrate that the monolauryl compound shown has optimal antibacterial
and antifungal properities relative to other cationic porphyrins with longer or shorter alkyl chains. AquaFrin is activated by
white light, with optimal activation occurring in blue wavelengths most readily transmitted in water.
AquaFrin shows low dark toxicity yet potent light toxicity to bacterial, fungal, and protozoal pathogens, including spores.
The selective toxicity is due to association with the anionic, lipophilic membranes of the microbial pathogens. For example,
Candida albicans has a light-activated MIC of 5 µg/ml. Concentrations as low as 0.2 µM, or only 0.3 g/1000 L tank, controls
Saprolegnia parasitica with 1-h treatments on two or three consecutive days. These concentrations are 100 to 1000-fold lower
than concentrations of chloramine-T, formalin, hydrogen peroxide, hydrogen peroxide-peracetic acid, copper sulfate, or potassium
permanganate solutions used as approved antimicrobials in aquaculture. Finally, AquaFrin is not a primary skin irritant
(Draize test, murine model) even at a dose of 300 µΜ, over 1000-fold higher than the concentrations used for decontamination
of tanks or races containing juvenile or adult fish.
This presentation will describe the latest results on optimization of wavelength and light flux to achieve selectivity and potency
for selected aquaculture pathogens. We will also present the newest evidence in support of environmental and food safety, as
well as target animal species safety and efficacy of this investigational new animal drug (INAD), which is being assembled with
the support of the Aquatic Animal Drug Approval Program and its affiliated laboratories.

EVALUATION OF MULTIPLE-BATCH CHANNEL CATFISH Ictalurus punctatus
PRODUCTION FED FOUR RECENTLY-DEVELOPED COMMERCIAL DIETS
Neil Pugliese, David Heikes, Carole Engle and Brian Bosworth
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
1200 N. University Dr.
Pine Bluff, AR 71601 USA
With prices for channel catfish feed soaring, feed mills have adopted new feed formulations in an attempt to lower feed costs.
This study compared survival, growth, FCR and net yield of four recently developed commercially available channel catfish
diets in a multiple-batch production scenario. On 8 May 2008, sixteen 0.1-ha ponds were stocked with 14,388 fingerlings/ha
(12.5 cm) and 4,308 head/ha (350 g) of carryover fish. The feeds used in this study were: 1) 32% corn gluten feed, 2) 32%
premium feed, 3) 32% standard feed, and 4) 24% premium feed. Each of the feed treatments was assigned randomly to four
ponds. The effect of the various feeds on processing parameters such as the headed-gutted weight, untrimmed fillet weight,
shank fillet weight, nugget weight, visceral fat content, and the fillet fat content were also compared.
At harvest (23-26 September 2008), the net yield of carryover fish fed varied significantly with the diet fed. The 32% premium
diet had 318 kg/ha greater yield than the 32% standard diet that had 191 kg/ha greater yield than the 32% corn gluten diet that
had 373 kg/ha greater yield than the 24% premium diet. The 32% premium and 32% standard feed treatments had similar
FCR’s that were significantly lower than the FCR’s of the 32% corn gluten feed and the 24% premium feed. There was no
difference in shank fillet yield among treatments but the catfish fed 32% corn gluten feed had significantly less visceral fat. A
partial budget analysis compared switching to the 32% standard or 32% corn gluten feed from 32% premium. The higher yield
from 32% premium feed more than offset its higher cost. It was more profitable to feed the more expensive 32% premium
feed.
2

2
RAPID AND SIMPLE DETECTION OF INFECTIOUS MYONECROSIS VIRUS (IMNV) BY
REVERSE TRANSCRIPTION-LOOP-MEDIATED ISOTHERMAL AMPLIFICATION IN
Penaeus vannamei
Teeranart Puthawibool*, Saengchan Senapin, Wansika Kiatpathomchai and TimothyW. Flegel
CENTEX Shrimp, Faculty of Science
Mahidol University
Rama 6 Road
Bangkok 10400, Thailand
nokky_pp@hotmail.com
Infectious myonecrosis virus (IMNV) has caused a slowly progressive disease with cumulative mortalities of up to 70% or
more in cultured Penaeus (Litopenaeus) vannamei in Northeast Brazil and Indonesia. Rapid detection of viruses by loop-
mediated isothermal amplification (LAMP) of genomic material with high specificity and sensitivity can be applied for diagnosis,
monitoring and control of diseases in shrimp aquaculture. Using an IMNV template, successful detection was achieved
after a 60-min RT-LAMP reaction using biotin-labeled primers followed by 5 min hybridization with an FITC-labeled DNA
probe and 5 min assay using a chromatographic lateral flow dipstick (LFD). Thus, the combined system of RT-LAMP and
LFD required a total assay interval of less than 75 min, excluding the RNA extraction time. The sensitivity of detection was
comparable to that of other commonly used methods for nested RT-PCR detection of IMNV. In addition to reducing amplicon
detection time when compared to electrophoresis, LFD confirmed amplicon identity by hybridization and eliminated the need
to handle carcinogenic ethidium bromide. The RT-LAMP–LFD method gave negative test results with nucleic acid extracts
from normal shrimp and from shrimp infected with other viruses including infectious hypodermal hematopoietic necrosis virus
(IHHNV), monodon baculovirus (MBV), a hepatopancreatic parvovirus from P. monodon (PmDNV), white spot syndrome
virus (WSSV), yellowhead virus (YHV), Taura syndrome virus (TSV), Macrobrachium rosenbergii nodavirus (MrNV) and
gill associated virus (GAV).

ENGINEERING OPTIMAL SEED PHOSPHORUS FOR GRAIN AND LEGUME USE IN FISH
FEEDS
Victor Raboy
USDA Agricultural Research Service
Aberdeen, Idaho 83210 USA
Victor.raboy@ars.usda.gov
There is growing interest in using cereal grain and legume products in aquaculture feeds, but this will require consideration
of seed phosphorus (P) amount and chemistry. P is an important nutrient that must be supplied to growing fish in adequate
amounts for optimal production, but high levels of P in aquaculture effluents represents an environmental hazard. Protein products
obtained from seeds are relatively high in P (perhaps ranging from 5 to 10 gm P kg -1 ) but at least 75% of this P is found as
phytic acid P (myo-inositol 1,2,3,4,5,6 hexakisphosphate), a form of P most fish do not efficiently utilize, and thus excrete.
One approach to dealing with the “phytic acid problem” has been to engineer low phytic acid genotypes of crop species. Multiple
“low-phytate” types have now been developed in maize (Zea mays L.), rice (Oryza sativa L.), wheat (Triticum aestivum
L.), barley (Hordeum vulgare L.), soybean (Glycine max L. (Merr.) and common bean (Phaseolus vulgaris L.). In normal seeds
(Fig. 1B), about 75% of seed total P is found as phytic acid P, less than 10% as inorganic P, and the remainder in “other” forms
of P. In most low phytic acid types (Fig. 1E), phytic acid reductions are matched by increases in inorganic P, so that seed total
P remains largely unchanged. Since most non-phytic acid P is nutritionally “available” to fish, available P is greatly increased
in low phytic acid types.
We are currently also screening for “high” and “low” seed total P genotypes. The genotype illustrated in Fig. 1C produces seed
that is high total P but the proportion of total P found as phytic acid P is normal. In contrast, Fig. 1D illustrates a high seed total
P genotype that is also low phytic acid. This seed would be very high in inorganic P and would have very high “available P”.
This probably would represent the ideal seed P amount and chemistry for use in aquaculture feeds. Our seed P genetics research
and aquaculture studies that evaluate these genotypes in feeds will be reviewed. This work will contribute to the NOAA-USDA
Plant Products in Aquaculture Working Group’s Strategic Research Plan Goal 3: “Enhance the inherent composition of crops to
provide a beneficial balance of bioactive compounds in order to optimize their use in aquafeeds for carnivorous fish.”
2

2
ULTRAVIOLET AND VISIBLE WAVELENGTH REFLECTANCE VALUES OF
AQUACULTURE PONDS AND THE ASSOCIATED USE BY DOUBLE-CRESTED
CORMORANTS
Andrew A. Radomski* and Paul Zimba
Harry K. Dupree-Stuttgart National Aquaculture Research Center
U.S. Department of Agriculture
Agriculture Research Service
P.O. Box 1050
Stuttgart, AR 72160 USA
Andy.Radomski@ars.usda.gov
Double-crested cormorants (Phalacrocorax auritus) are a frequent and major migratory avian predator on channel catfish
(Ictalurus punctatus) and other aquaculture species during the winter months throughout southeastern USA. Cormorant
numbers and foraging activity indicate a nonrandom distribution within the aquaculture industry. To address the hypothesis
that cormorants are selecting ponds based on reflectance values, we quantified bird use at each 3 spatially isolated commercial
aquaculture facilities. Although cormorant movements, numbers, and occurrence within the aquaculture production region
have been addressed, no studies have focused on the potential mechanisms utilized by these avian predators in selecting
foraging sites within the aquaculture high density prey-based systems.
The focus of this study was to determine if cormorants are associated with specific reflectance spectrum within channel catfish
ponds. Four ponds classified as ‘preferred’ and another four ponds were classified as ‘avoided’ based on observing cormorant
numbers for three months prior to measuring pond reflectance; >100 hrs observed per farm on three commercial catfish farms.
Reflectance and pond turbidity samples were measured every 10 days from January to March 2007; corresponding with peak
number of over-wintering cormorants within the region. Pond samples were measured at the subsurface (1 cm) and secchi
depths to determine spectral reflectance and turbidity. The overall pond surface reflectance model explained 54 per cent of the
variation within high and low bird use pond. The surface reflectance values between 380-410 nm (ultraviolet) and 540-680
nm, 750-760 nm and 800 nm (visible) were significantly different between the high bird use and low bird use. The secchi depth
reflectance showed similar patterns to the subsurface findings. The reflectance wavelengths may serve as a partial cue that
cormorants utilize in selecting particular commercial catfish ponds. The association of particular reflectance and cormorant use
will serve as the building block for creating predictive models and though cormorant numbers and movement patterns may not
change, this improved understanding of avian depredation at commercial aquaculture facilities will greatly impact the overall
effectiveness and cost to implement management strategies.

EFFECTS OF PREBIOTICS ON NUTRIENT DIGESTIBILITY OF A SOYBEAN- MEAL-
BASED DIET BY GOLDFISH Carassius auratus
Thiago Raggi*, Gary Burr, Brian Ray, Alejandro Buentello and Delbert Gatlin III
Department of Wildlife and Fisheries Sciences
Texas A&M University
College Station, Texas 77843 USA
thiago_82@tamu.edu
A combination of handling stress and suboptimal environmental conditions can result in high mortality on baitfish production.
Different dietary supplements may potentially reduce these losses by increasing immunological response, resistance to disease and
handling stress. Prebiotic compounds comprise one such group of supplements and are defined as non-viable food ingredients that
are selectively metabolized to favor beneficial intestinal bacteria, which may confer various desirable effects including enhanced
disease resistance and nutrient availability to the host organism. Such effects have been observed in some fish species such as red drum
(Sciaenopos ocellatus) and hybrid striped bass ( Morone chrysops x M. saxatilis) as well as golden shiner (Notemogonus crysoleucas).
This study examined the effects of four prebiotics on digestibility of soybean-meal-based diets. The four prebiotics were
GroBiotic ® -A (consisting of a mixture of partially autolyzed brewers yeast, dairy ingredient components and dried fermentation
products), mannanoligosaccharide (MOS), galactooligosaccharide (GOS), and fructooligosaccharide (FOS), each added to a
basal diet at 1% by weight. The diets were formulated so that 50% of the protein was provided by soybean meal and the other
50% was from menhaden fishmeal. Chromic oxide was added to the diets at 1% as an inert marker. Each diet was fed to adult
goldfish in duplicate 110-L aquaria for a total of 8 weeks, and manually stripped to obtain fecal material approximately 10 hours
after feeding on four different occasions. The dried fecal material from each aquarium was pooled over time and analyzed for
protein, lipid, organic matter and chromium in order to compute coefficients of digestibility. Analyses are currently being conducted.
2

2 0
EFFECTS OF FISHMEAL VERSUS SOYBEAN-BASED FEEDS AND SOLIDS REMOVAL
BY SETTLING TANKS AND TILAPIA ON HIGH-DENSITY SHRIMP Litopenaeus vannamei
PRODUCTION IN BIOFLOC CULTURE SYSTEMS
Andrew J. Ray*, John W. Leffler, Gloria Seaborn, Beth Thomas, Jesus A. Venero, Luis Vinatea and Craig L. Browdy
SCDNR - Waddell Mariculture Center
211 Sawmill Creek Rd.
Bluffton, SC 29910 USA
raya@dnr.sc.gov
Biofloc-based culture systems function as an environmentally friendly, cost-effective method of growing shrimp (Litopenaeus
vannamei) at high density. The microbial communities, largely contained within biofloc particles, are responsible for nutrient
cycling, system stability, and growth-enhancement benefits for shrimp. Regulating the abundance and composition of these
microbes is an important step in maintaining reliably productive systems. In addition, the feeds used in biofloc systems should
be cost-effective, eco-friendly, and provide optimal nutrition for accelerated shrimp growth. We explored these requirements
by applying three biofloc solids management tactics along with two feed formulations in high-density culture units. System
dynamics and shrimp performance parameters were evaluated in light of these treatments.
Shrimp were stocked at 300/m 2 in 3.35m diameter, 6.3m 3 outdoor tanks. A high quality, soybean meal-based diet and a traditional,
fishmeal-based diet were each administered to half of the tanks. Among tanks receiving each diet, one third had solids
removal in the form of a settling chamber, one third had water diverted through a tilapia tank, and the remaining third received
no additional management. The combination of solids removal tactics and feed types created six unique treatments; each was
replicated four times. Differences in biofloc nutrition were measured with proximate analysis and fatty acid assessments.
System dynamics were assessed by measuring turbidity, total and volatile suspended solids, water quality parameters, percent
light penetration and community respiration. Microbial dynamics were monitored with light microscopy observations and fatty
acid biomarkers. Shrimp performance was characterized by mean individual weight, biomass yield, feed conversion ratio, and
survival. Shrimp quality was determined using a double-blind taste test and by testing product shelf-life.
The two diets led to differences in shrimp growth and feed conversion ratios. Differences in the nutritional profiles of biofloc
generated by the two diets were documented and these correlated with disparities in shrimp flesh nutrition and quality. Removal
of suspended solids by both the settling chambers and the tilapia increased light availability in tanks and reduced the
occurrence of potentially harmful cyanobacteria. Settling chambers reduced the abundance of bacteria and zooplankton while
tilapia reduced algae from systems. Both settling-based and tilapia-based solids removal increased system stability by reducing
oxygen demand and enhanced shrimp growth and survival compared to the non-cropped systems. This project demonstrates
that suspended solids management techniques provide benefits for shrimp production in high-density biofloc culture. The
study also shows that a carefully formulated plant-based diet can produce shrimp production results equivalent to those of a
traditional, fishmeal diet.

EVALUATION OF SYNTHETIC ASTAXANTHIN AND ALFALFA MEAL AS A POTENTIAL
DIET ADDITIVE FOR COLOR ENHANCEMENT OF KOI Cyprinus carpio
Andrew J. Redden*, Boris Gomelsky, Carl D. Webster and Robert M, Durborow
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
Andrew.redden@kysu.edu
Koi are a colorful ornamental variety of common carp Cyprinus carpio and are members of the family Cyprinidae. Aquaculturists
have become interested in koi because of their high-value as an ornamental fish. Ornamental fish species represent the most
valuable aquaculture products in the U.S. with a retail value over one billion dollars annually. Due to increased popularity and
production of koi in the US, wholesale buyers are now demanding higher quality fish with higher levels of pigmentation. Since
skin pigmentation is a leading trait for koi enthusiast, some studies have attempted to enhance coloration. Thus, development of
improved formulated practical koi diets that enhance color would be an economic benefit to koi aquaculture enthusiasts. Extensive
research has been conducted to enhance the color and therefore the value of ornamental fish by using carotenoids as feed
additives. Synthetic astaxanthin and several other sources of carotenoids have been proven to increase color in koi. However,
the cost of these carotenoids can increase the cost of producing diets up to 100%. Thus, less costly carotenoid sources must
be evaluated for color enhancement of ornamentals. Although deficient in lysine, alfalfa meal is recommended for use at an
inclusion rate of 15-20% in diets for grass carp. Alfalfa meal is a cost effective source of carotenoids and protein and should be
evaluated for its effect on koi growth and coloration. The objective of this research study is to determine the optimal inclusion
rates of synthetic astaxanthin with or without the addition of 15% alfalfa meal in diets for juvenile koi for color enhancement
without reducing growth performance.
A 10-week study using 21, 114-L glass aquaria stocked at 30 fish per aquarium will be conducted on 2 to 3 gram juvenile koi.
Seven experimental diets will be evaluated with three replicate tanks per experimental diet. Fish will be fed seven experimental
diets to satiation twice a day. The basal control diet will contain fish meal, soybean meal, wheat gluten meal, vitamin mix,
mineral mix, vitamin C, choline and di-calcium phosphate. Three experimental diets will be formulated similar to the basal diet
with the addition of astaxanthin while the other three diets will contain the same amount of astaxanthin as the previous diets
with the inclusion of 15% alfalfa meal (Table 1). Color enhancement will be evaluated every 10 days using a portable Minolta
Chroma Meter CR-400 calibrated towards a white standard. Results will be discussed.
Table 1. Astaxanthin amounts and alfalfa
meal % for experimental diets.
Diet
Astaxanthin
(mg/kg)
Alfalfa Meal
(%)
Basal 0 0
1 20 0
2 40 0
3 60 0
4 20 15
5 40 15
6 60 15
2 1

2 2
ASSESSMENT OF GROWTH AND SURVIVAL ON LARVAL COMMON SNOOK, Centropomus
undecimalis, FED ROTIFERS ENRICHED WITH ARACHIDONIC ACID
Matthew J Resley*, Nicole Rhody and Kevan Main
Mote Marine Laboratory
Center for Aquaculture Research and Development
Sarasota, Florida 34236 USA
resleymj@mote.org
Efforts to develop the culture technology for common snook (Centropomus undecimalis) have been ongoing for more than 20
years. Populations of this popular sportfish have been declining due to human influences. Because of their decreasing population
size research is underway to evaluate the potential to increase their populations using stock enhancement. Though there
have been some successes in producing this species for stock enhancement, rearing technology for common snook is still in its
infancy. Currently at Mote Marine Laboratory we are trying to increase culture efficiency and technology with work on captive
broodstock, and larval and juvenile rearing practices. Preliminary data from other studies at our facility revealed a possible
deficiency in arachidonic acid (ARA) in the captive broodstock and subsequently their eggs. This essential fatty acid plays an
important role in development and could be important through the larval phase. This project was set up to assess whether ARA
enrichment of rotifers can increase growth and survival of snook larvae.
The control treatment was our standard protocol for rotifer enrichment. The control enrichment used was Algamac 3050 tm and
protocols were obtained from the manufacturer (0.2 g/L of enrichment for 12 hr). The two experimental enrichments utilized
Algamac-ARA tm . The two treatments were a 10 and 20% replacement of the Algamac 3050. The 10% replacement was the
manufacturers suggested replacement, and the 20% replacement was used to evaluate higher levels of ARA in the larval feed.
Rotifers were harvested from the continuous culture system on site one day before feeding. They were held in 18 L PVC buckets,
where they were enriched. Three samples of enriched rotifers were also obtained from each treatment for lipid analysis.
Rotifers were maintained at 5/ml throughout the study. The study was initiated when larvae were 3 days post-hatch (day 1 of
the study).
Larvae for this experiment were obtained from captively held broodstock at Mote Aquaculture Park in Sarasota, Florida. Larvae,
1 DPH, were stocked into 12 100-L black conical experimental tanks at a density of 100 larvae per liter. To assess larval
survival from the tank stocking, six 1 L beakers were stocked with 20 larvae each, and survival was recorded the day following
the move. The remaining larvae were left in hatchers to be sampled two days later for initial larval measurements; standard
length, mean dry weight, and fatty acid profiles. There were 20 larvae removed for standard length, 50 for dry weight measurements,
and 3 x 200 for lipid analysis. Dry weights were obtained by rinsing the larvae with DI, and placed in a drying oven
at 60°C for 24hr. The larvae sampled for the lipid analysis were rinsed as described above for weighing, and placed in glass
vials and stored in a -80°C freezer. This procedure was repeated at day 7 (midpoint) and day 15 (end of study), with fish being
removed from each tank. Remaining fish at the end of the study were sieved and counted to assess tank survival.

EFFECT OF ZOOPLANKTON DENSITY ON GROWTH OF LARVAL AMERICAN SHAD
Alosa sapidissima: AN EXPERIMENTAL APPROACH
Kenneth L. Riley, Samantha M. Binion, Travis T. Williams and Anthony S. Overton
East Carolina University
Department of Biology
Greenville, North Carolina 27858 USA
klr1011@ecu.edu
Widespread declines in stocks of American shad Alosa sapidissima along the Atlantic Coast have been attributed to overfishing,
decrease in water quality, and loss of habitat. Recent surveys in North Carolina suggest that stocks are continuing to decline
despite extensive management and stock enhancement efforts. We hypothesize that the precipitous decline in American shad
reflects the availability of suitable nursery habitat and food resources for early life stages in Roanoke River and Albemarle
Sound. The goal of this study was to conduct laboratory trials to evaluate the effect of prey density on growth and survival of
American shad. Larvae were reared at 25 C from 12 to 20-d posthatch in five treatments: (1) starvation; (2) low-prey, which
simulated natural prey densities in Roanoke River; (3) medium-prey, which simulated natural prey densities in adjacent riverine
systems; (4) high-prey, and (5) Artemia nauplii. The latter treatments simulated prey densities typically used in hatchery
operations. Wild zooplankton collected from local waters were used as prey. Zooplankton were identified and measured. Zooplankton
primarily consisted of cladocerans (29%), copepod nauplii (25%), rotifers (24%), and copepod adults (19%). Larval
survival was 35 ± 7% and was not significantly different among treatments. The highest survival was observed with fish fed
high concentrations of prey (46 ± 18%) followed by Artemia (40 ± 16%). The lowest survival was observed with starved fish
(22 ± 12%). Length-specific growth rates (G l ) determined from total length measurements were 0.069 for Artemia, 0.056 for
high-prey, 0.029 for medium-prey, 0.017 for low-prey, and 0.009 for starved treatments. Larval growth as a function of length
was not significantly different between Artemia and high-prey; however, these treatments were significantly higher than the
lower prey densities (ANOVA; P < 0.0001). Weight-specific growth rates (G w ) determined from dry weight measurements
were significantly higher for Artemia (G w = 0.851; P < 0.0001) and remained relatively low for all other treatments (G w =
0.617). Evidence from this research shows that zooplankton densities common to Roanoke River and Albemarle Sound are
not optimal for growth of American shad. Releases of hatchery-reared fish should be timed to coincide with natural peaks in
zooplankton production.
FIGURE 1. Growth (mean
± S.E.) of American shad
larvae reared at 25 C and
cultured in 12-L tanks (3
fish L -1 ). Prey densities
ranged from 1.0 x 10 3 to
1.0 x 10 6 individuals m -3 .
2 3

2 4
effectS of 17α-metHYLteStoSterone on GonADAL SeX DifferentiAtion in
SPOTTED SAND BASS (Paralabrax maculatofasciatus)
Martín O. Rosales-Velázquez*, Rosa I. Ochoa-Báez, Wilfrido M. Contreras –Sánchez, José L. Ortiz-Galindo,
Tanos Grayeb-del Alamo and Víctor Carrasco-Chávez.
Laboratorio de Biología Experimental
Centro Interdisciplinario de Ciencias Marinas-IPN
La Paz, B.C.S. Mexico 23096
mrosales0400@ipn.mx
Knowledge of a species for aquacultural purposes includes diverse factors that must be studied and understood to detail. One of
these factors is the way that sexual determination takes place in a species, and the plasticity of manipulating sex differentiation
in order to obtain single sex populations. Production of monosex populations has two objectives: to obtain the sex with faster
growth and avoid reproduction to direct energetic budget to somatic growth. Due to its protoginic condition, for the spotted
sand bass (Paralabrax maculatofasciatus) we have proposed the use of all male populations.
In the present study, oral administration of 17 α-Methyl testosterone (MT) was used to masculinize the fish. Apparently, all
doses tested (15, 30 and 45 mg/Kg) were too high, impeding gonadal development and resulting in sterile fish (figure 1). More
than 50% of the fish had no gonads. Significant differences were found for gonadosomatic index (GSI), being significantly
larger for the fish of the control group (Figure 2). During early development the activity of the enzyme aromatase was significant,
clearly indicating female development.
In conclusion, all treatments and times used were sufficient to attain the raised objective consisting in avoiding females in the
population, and consequently impeding reproduction. Fish in the control group were masculinized presumably due to a leakage
of MT into the Recirculating System.

CORN DISTILLERS DRIED GRAINS WITH SOLUBLES (DDGS) AS AN ALTERNATIVE
PROTEIN SOURCE: OPPORTUNITIES AND CHALLENGES FOR AQUACULTURE
Kurt A. Rosentrater
USDA-ARS, North Central Agricultural Research Laboratory
2923 Medary Ave.
Brookings, SD 57006 USA
kurt.rosentrater@ars.usda.gov
As the demand for, and cost of, aquaculture production increases, it
is becoming increasingly imperative that viable alternative feeds be
developed and utilized. Corn-based distillers dried grains with
solubles (DDGS) is a feed material produced by the fuel ethanol
industry. As the biofuels industry continues to grow (Figure 1), this
coproduct should be given greater consideration as a protein source
for aquafeeds. The goals of this presentation thus include a discussion
of processes that are used to produce DDGS, physical and chemical
properties thereof, and challenges encountered when using this feed
ingredient. The information discussed herein falls within the auspices
of Goal 5 of the Plant Products in Aquafeed Working Group Strategic
Research Plan.
Converting corn to ethanol (Figure 2) consists of grinding, cooking,
liquefying, saccharifying, fermenting, and distilling. Nonfermentable
components are removed as whole stillage, centrifuged to remove
water – which is then evaporated to produce condensed distillers solubles
(CDS), and then is recombined with the centrifuge solids and
dried to produce DDGS. Each bushel of corn (56 lb) will result in
nearly 2.8 gal of ethanol, 18 lb of CO 2 , and 18 lb of DDGS.
Typically, on a dry basis, DDGS are approximately 30% protein, 10%
lipid, over 30% neutral detergent fiber, and up to 10% starch. Composition,
however, can vary considerably between plants (Table 1), and
within a single plant over time.
There are many issues associated with using DDGS.
Some of the most pressing include variability in nutrient
content and quality; lack of an industry-wide quality grading
system; inconsistent product identity and nomenclature;
the large quantities of energy required to remove water and the high cost of energy; moving DDGS to diverse and distant
markets; potential mycotoxin contamination; and international marketing and export challenges.
2

2
ORGANIC CARBON AND NITROGEN REMOVAL FROM GARFISH AQUACULTURE
USING SEQUENTIAL BATCH REACTOR - A LABORATORY SCALE STUDY
Dhritikshama Roy* and Raj Boopathy
Department of Biology
Nicholls State University
Thibodaux, LA 70310 USA
Royd425@its.nicholls.edu
Garfish is a fresh water fish that can tolerate moderate range of salinity. Research is underway at Nicholls State University to
grow garfish under various salinity levels because of the availability of saline water. Aquaculture of garfish is in experimental
stage in Louisiana. The experimental aquaculture system involves feeding garfish with 40% protein diet. This higher protein
diet resulted in the accumulation of high concentration of dissolved organic carbon (~ 6500mg/L), ammonia-N (~ 600 mg/L),
nitrite-N (~ 400 mg/L)) and nitrate (~650 mg/L)) in garfish aquaculture system.
The objective of this study was to treat the garfish aquaculture waste water using Sequential Batch Reactor (SBR). Sequential
batch reactor (SBR) is an activated sludge treatment with a combination of variable period of aerobic and anaerobic modes.
The experiment used garfish waste water of three different concentrations of salt – 4.0 ppt, 8.0 ppt and 12.0 ppt. Garfish waste
without salt served as control. Performance of SBR showed significant removal of dissolved organic carbon and total nitrogen
within 15 days with 11 days of aerobic condition followed by 2 days of anaerobic and 2 days of aerobic modes for various
salinity garfish waste water. The removal rate was concentration dependent. The quantitative effect of heterotrophic bacteria in
garfish aquaculture was also noticeable during waste removal process through SBR.

EFFECT OF TEMPERATURE AND SALINITY ON GROWTH AND SURVIVAL OF POST-
LARVAL AND JUVENILE NATIVE AND NON-NATIVE SHRIMP IN ALABAMA
Luke A. Roy*, D. Allen Davis, Herbert E. Quintero, Jessica Jacquay, Patricio Paz and Daranee Sookying
Department of Fisheries & Allied Aquacultures
Auburn University
Auburn, AL USA
royluke@auburn.edu
Very little is known about the interaction of temperature and low salinity as it relates to survival and growth of post-larval and
juvenile shrimp. Both temperature and salinity are known to influence the growth potential and survival of shrimp. In west Alabama,
farmers suspect low water temperatures (in spring and fall), in conjunction with low salinity and suboptimal ionic profiles,
might be responsible for reduced survival and production at harvest. Likewise, the modeling of growth of native shrimp
species either in the wild or under aquaculture conditions cannot be predicted as there is also limited data on these species. In
order to determine the influence of temperature and salinity on post-larval and juvenile L. vannamei and F. duorarum, a series
of bioassays were conducted at the E.W. Shell Fisheries Research Station (EWS) in Auburn, Alabama and Claude Peteet Mariculture
Center (CPMC) in Gulf Shores, Alabama. Litopenaeus vannamei were obtained from Shrimp Improvement Systems in
the Florida Keys, Florida and held at EWS and CPMC until the commencement of the bioassays. At EWS 15 post-larvae (PL 13
and PL 20 ) were placed in twenty buckets containing 2 Liters of water with an initial salinity of 12 ppt and a constant temperature
of 19.3°C (PL 13 ) and 19.6°C (PL 20 ) Using a drip irrigation system, freshwater was added to each bucket at a rate of approximately
1 gallon per hour. Each bucket was equipped with an airstone supplied with aeration from a regenerative blower. The
target salinities for both bioassays were 12(control), 4, 2, 1, 0.5, and 0.2 ppt. Salinity was reduced at a rate of 4 ppt per hour
until 4 ppt. After reaching 4 ppt the rate of salinity reduction was reduced to 1 ppt per hour until 1 ppt. After reaching 1 ppt, one
hour was allowed to reach 0.5 ppt and another hour to reach 0.2 ppt. Survivals of PL 13 shrimp (48 hour) were not significantly
different among 12, 4, and 2 ppt treatments (83.3-91.7%). However, 1, 0.5, and 0.2 ppt treatments had significantly lower (P
< 0.05) survivals (0-18.3%) when compared to 12, 4, and 2 ppt. Survivals of PL 20 shrimp (48 hour) were not significantly different
among 12, 4, 2, and 1 ppt treatments (95-100%). However 0.5 and 0.2 ppt treatments had significantly lower (P < 0.05)
survivals (0-41.4%) when compared to 12, 4, 2, and 1 ppt. The differences in survivals among PL 13 and PL 20 shrimp indicate
an age effect with respect to salinity tolerance following a 48 hour acclimation. The same bioassays were repeated with L. vannamei
at CPMC using PL 11 (22.6°C constant temperature) and PL 20 (17.7°C constant temperature) with similar target salinities
(30, 4, 2, 1, 0.5, 0.2 ppt). Shrimp were stocked into 15 buckets (3 replicates per treatment). Survivals were similar to the bioassays
conducted at E.W. Shell. Similar experiments were also conducted using F. duorarum at both EWS and CPMC. Results
indicate that age is an important factor to consider when acclimating post-larval shrimp to low salinities.
2

2
EFFECT OF FEEDING RATE ON SURVIVAL AND GROWTH OF L. vannamei REARED IN
LOW SALINITY WATERS OF WEST ALABAMA
Luke A. Roy*, D. Allen Davis and Greg N. Whitis
Department of Fisheries & Allied Aquacultures
Auburn University
Auburn, Alabama 36849 USA
royluke@auburn.edu
Feeding costs have sky-rocketed worldwide in the last year, negatively impacting shrimp farmers. In most areas of the world,
50-60% of production costs in shrimp farm operations are directly attributed to the feed. The increasing cost of feed and fuel
combined with declining market prices for shrimp is making it more difficult to maintain adequate profit margins in the shrimp
industry. In west Alabama, shrimp farmers are located long distances from feed mills and are also forced to pay high shipping
fees for their feed. Due to these reasons, shrimp farmers are always interested in saving money on feed. The low salinity waters
utilized for shrimp culture in west Alabama are quite productive, providing additional natural food items to shrimp throughout
the production season. In order to determine the possibility of reducing feeding rates, an experiment was devised at Greene
Prairie Aquafarm in west Alabama. The experimental system consists of a series of 600 L tanks fed with water from a low salinity
shrimp production pond. Each tank is equipped with two air stones supplied with air from a regenerative blower. Dissolved
oxygen, temperature, and pH were measured twice daily throughout the experimental period, while salinity, ammonia-nitrogen,
and nitrite nitrogen were measured weekly. Thirty tanks were stocked with 20 shrimp (0.80 g initial weight) per tank. A production
diet (Rangen Inc. 40% protein 5% squid) was utilized as the diet for the experiment. Shrimp were offered the diet at
a rate of 60%, 70%, 80%, 90%, 100%, and 110% of a pre-determined daily ration resulting in 6 treatments with 5 replicates.
The 100% treatment represents a typical amount of feed based on an assumed 1.5 g of growth per week and an FCR of 1.25.
The experiment is presently ongoing and will be harvested after 10 weeks. At harvest, growth, survival, percent weight gain,
and FCR will be determined. Results from this experiment will help determine whether west Alabama farmers can reduce feed
rates and rely on the primary productivity of the pond to supply a portion of their nutritional needs.

NOAA-USDA JOINT INITIATIVE ON ALTERNATIVE FEEDS FOR AQUACULTURE
Dr. Michael J. Rubino
NOAA Aquaculture Program
1315 E-West Hwy.
SSMC#3, 13th Fl., Room 13117, Mail Code: F
Silver Spring MD 20910 USA
Michael.Rubino@noaa.gov
Fish meal and fish oil are important components in the feeds for many farm-raised species, from pigs and poultry to farmed
fish. As ingredients in aquaculture feed, fish meal and fish oil supply essential amino acids and fatty acids required for normal
growth for cultured species, including carp, salmon, tilapia, trout, catfish, shrimp and others. Fish meal and oil also help maintain
the important human health benefits of seafood. However, the relatively high cost of fish meal and fish oil – and growing
pressure on the wild fisheries that supply the fish meal and fish oil – are adding up to make alternative feeds one of the top
issues facing the global aquaculture industry, fueling research on suitable alternative feed ingredients.
In the United States and elsewhere, studies are underway to better understand the nutritional requirements of fish and shrimp
and to evaluate the use of alternative dietary ingredients in aquaculture feed. Potential alternatives already in use include soybeans,
barley, rice, peas, and other crops along with canola, lupine, wheat gluten, corn gluten, various plant proteins, algae, and
seafood processing co-products. To help maintain the human health benefits of eating seafood, suitable alternatives with marine
nutrients such as long chain omega 3 fatty acids, are needed.
On a global scale, significant improvements have been made in reducing the reliance on fish meal and fish oil for feeds for
many cultured species. NOAA and other federal agencies play a vital role in that research and the transfer of that technology
to industry. In November 2007, the National Oceanic and Atmospheric Administration (NOAA) in partnership with the U.S.
Department of Agriculture (USDA) launched the Alternative Feeds Initiative to accelerate the development of alternative feeds
for aquaculture. This presentation will address the major milestones of the initiative to date, highlight ongoing research and
identify new priority areas.
The overall purpose of the NOAA-USDA Alternative Feeds Initiative is to identify alternative dietary ingredients that will reduce
the amount of fishmeal and fish oil contained in aquaculture feeds while maintaining the important human health benefits
of farmed seafood. Ultimately, the initiative will lead to the commercialization of alternatives for some species which will result
in reduced dependence on marine fish resources by feed manufacturers and seafood farmers worldwide. NOAA is partnering
with the USDA’s Agricultural Research Service and Cooperative State Research, Education, and Extension Service on the initiative,
which will build on ongoing USDA and NOAA research to identify alternative protein and oil sources for aquaculture
feeds.
2

300
MIXTURE DESIGNS FOR FEED FORMULATION AND FISH NUTRITION STUDIES
Kari Ruohonen* and Juhani Kettunen
EWOS Innovation
N-4335 Dirdal
Norway
kari.ruohonen@ewos.com
Mixture designs are natural choices for problems on fish feed formulation or fish nutrition. Usually the factors affecting responses,
i.e. the formulations, have constraints that, if not accounted for, will confound the effect estimates. The main constraint
always present for a formulationfollows from the fact that the total amount of ingredients of the mixture must sum up to one
hundred percent. More constraints normally exist. Existing knowledge of fish nutrition introduces minimum/maximum constraints
for main nutritive substances. Furthermore, also production technology and prices of raw materials set up constraints
for technical and economical feasibility of the feed mixtures. In the paper, we will demonstrate the use of mixture models by
summarizing our experience from case studies. We will show how a typical mixture problem is designed and analyzed. Principally,
the problem solving is a process of consecutive, systematic change of scales connected with linear models. The scaling
process is necessary for at least the following four reasons. First, it is used to take into consideration the constraints of the problem.
Second, the scaling is needed to give an optimal statistical weighing of the ingredients in the mixture. Third, the scaling
greatly improves the estimation accuracy. Fourth, the scaling saves money and effort, because it constraints the experimental
effort into a feasible region and abolishes idle levels of ingredients. The mixture designs and models are a great improvement
to the present status of feed formulation and nutrition studies. However, we demonstrate also some challenges in their use. One
issue in applying mixture designs is whether the component axes should be based on raw ingredients or nutrient composition.

EFFECTS OF TEMPERATURE REGIME ON NITROGEN DEGRADATION: LABORATORY
AND MODELING RESULTS
Milton Saidu*, Steven G. Hall, Ron Malone and Terrence Tiersch
Biological and Agricultural Engineering
LSU AgCenter
Baton Rouge, LA, 70803 USA
msaidu1@lsu.edu
Temperature is a factor in growth of aquatic species. Previous work had considered how temperature may affect nitrogen
degradation in biofilters and other components of aquaculture systems. This work used a series of laboratory (40 liter) and
pilot scale (250 liter) temperature controlled tank systems (Figure1) to assess effects of different steady state and variable temperature
regimes on nitrogen degradation. A series of protocols were used to assure controlled conditions, including unique
heat exchangers, computer controlled heating and cooling and cleaning and measurement protocols which provided enhanced
replicability of experiments.
Steady state temperatures of 13, 20 and 30 each showed significant differences (Figure 2) in ammonia degradation rates. Diurnal
sinusoidal 20±3 and 30±3 temperature regimes also revealed significant differences and suggest that this level of variation
may enhance, not limit degradation. Step input studies provided similar evidence, while mixing beads acclimated at different
temperatures from 13 to 30C appeared to enhance degradation. Modeling studies helped clarify these results and provided
predictive tools for future design and management of biofiltration systems in controlled or natural settings.
301

302
THE SURVEY OF FISH CONSUMPTION ATTITUDE OF NUTRITION EXPERTS IN IRAN
Salehi, H.
Assistant Professor
Iranian Fisheries Research Organization, No., 297
Fatemi Ave.
P. O. Box: 14155-6116
Tehran, Iran
hsalehi_ir@yahoo.com
The wide relationship of nutrition experts in general and their roles to change the consumption attitude of the people are very
important. Though, the survey of fish consumption attitude of nutrition experts may used to promote fish extension planning
and policy making.
144 nutrition experts were selected and interviewed on September 2005. The results showed 64% of respondents were believed
the effect of promotion and this result was significantly related with age (P

ARACHIDONIC ACID (20:4n-6) EFFECT ON REPRODUCTIVE PERFORMANCE OF A
TROPICAL SILVERSIDE, Menidia estor
Rosa L. Salgado-García * , Antonio Campos-Mendoza, Jorge Fonseca-Madrigal, Danitizia A. Guerrero-Tortolero,
Carlos A. Martínez-Palacios, Ilie S. Racotta, Mathiew Wille, Patrick Sorgeloos and Elena Palacios 1
Aquaculture Program
Centro de Investigaciones Biológicas del Noroeste
S. C., CIBNOR
Mar Bermejo 195
Palo de Santa Rita .C.P. 23090
La Paz, B. C. S. México.
rsalgado@cibnor.mx
epalacio@cibnor.mx
Arachidonic acid (ARA) modulates reproduction, immune response, stress tolerance, type II prostaglandin synthesis and has
been reported to affect egg and larval quality in several marine temperate species. Menidia estor is an endemic freshwater fish
from Mexico that has excellent growth during culture. However, cultured organisms have poor egg and larval quality. A previous
study showed low ARA levels in cultured fish eggs compared to wild fish eggs. The aim of this study was to evaluate the
effect of supplementing ARA to broodstock on reproductive performance.
Two-y fish (36±1g) under standard culture conditions
(3000L, 0 psu, 17±1˚C, 18L:6O) were fed enriched adult
Artemia (1% fish weight day -1 ) with three different ARA
(ICES, 1997) levels (0, 1 and 8%) complemented with
pellet (3% weight day -1 ) during 70 days, in triplicate
(n=21, 1♀:2♂). ARA levels in the diet (Table 2) affected
growth but not survival or hepatosomatic index (HSI).
8%-ARA diet increased gonadosomatic index (GSI) in
males but not in females. Fecundity was higher in fish fed
8%-ARA while egg diameter was smaller in fish fed 0%-
ARA. Numbers of spawns, sperm density, fertilization
and eclosion rates were similar between the three ARA
diets (Table 1). No significant differences were found
in relation to diet in the fatty acid of membranes in the
gonadas (Table 3). In spite of a very low quantity of
22:6n-3 in the enriched Artemia, the concentration of this
fatty acid in membrane of gonads was very high, particularly
in females. Enrichment with ARA increased some
production variables.
303
Table 1. ARA effect on reproductive performance of M. estor. Data
expressed mean ± standard error (P

304
SUMMARY OF RECENT RESEARCH FINDINGS FROM THE TEXAS AGRILIFE
RESEARCH MARICULTURE LABORATORY AT FLOUR BLUFF, CORPUS CHRISTI
Tzachi M. Samocha*, Josh S. Wilkenfeld, Timothy C. Morris and Terry R. Hanson
Texas AgriLife Research
Shrimp Mariculture Research Facility
4301 Waldron Road
Corpus Christi, TX 78418 USA
t-samocha@tamu.edu
Traditional intensive shrimp farming practices call for high water exchange to maintain adequate water quality in grow-out
ponds. Also, release of nutrient-rich effluent waters into receiving streams can have a negative impact on the environment.
These practices raise concerns over the sustainability of shrimp industry. Beside the environmental issues, water exchanges
have been implicated in cases of viral disease outbreaks and massive crop losses. Use of super-intensive grow-out systems with
high quality seed stock under biosecure conditions and limited or no water exchange can minimize crop losses and negative
environmental impact.
The presentation summarizes recent findings associated with operating super-intensive nursery and grow-out raceway systems
for culture of the Pacific white shrimp, Penaeus vannamei, at the AgriLife Research Mariculture Lab under limited and/or no
water exchange. Special emphasis will be placed on water reuse, changes in key water quality indicators, nutrient recovery, feed
utilization, shrimp performance, and economic viability of these systems.

GROWTH RATE OF JUVENILE AMAZONIAN PAICHE Arapaima gigas, FED FORAGE
FISH OR PRACTICAL DIET AND RAISED UNDER CAPTIVITY: IMPLICATIONS FOR ITS
COMMERCIAL PRODUCTION
Manuel Sandoval*, Juan Lao, Beatriz Sandoval and Wilson Castillo
Centro de Acuicultura Tropical, Facultad de Zootecnia
Universidad Nacional Agraria de la Selva
Tingo María, Perú
msandtm@msn.com
Arapaima gigas, is one of the largest freshwater fishes of the Peruvian Amazon basin. This fish is known as paiche in Peru or
pirarucu in Brazil and it can grow up to 3m in length and can weigh over 200kg. The meat has an excellent nutritional profile
in terms of protein, essential fatty acids and amino acids. Because of overfishing, paiche is listed by the Convention on International
Trade in Endangered Species (CITES). This has prompted the need to raise paiche in captivity to preserve the species
in their natural habitat and establish a sustainable production for human consumption. The present study was conducted to
investigate whether feeding a commercial practical diet to juvenile paiche would improve growth rate more efficaciously than
a forage diet.
A 12-week trial was conducted with juvenile (420g) paiche. Two experimental groups of 24 animals each were established:
1) forage fish diet Cichlassoma amazonarum (FF), and 2) commercial practical fish diet with 40% protein (PD). The animals
were raised in cages (n=8/cage) and placed into an artificial pond. The feeding regime was 2.5% of body mass and administered
twice daily; water quality was checked three times per week. After 12 weeks, A. gigas fed a FF had significantly (P < 0.01)
higher growth rate compared to A. gigas fed the PD (Table 1).
30

30
STUDY OF CAPABILITY OF BIVALVE Anodonta cygnea IN ORGANIC COMPOUNDS OF
WATER INDIRECT REMOVAL
Lalik Sarikhani, Arash Javanshir, Masoud Ounagh and Abdoolah Haghpanah
Grad.Student of Fishery
Faculty of Natural Resources
University of Tehran
In this study, filtration rate of water born- Nitrogen and phosphorus was evaluated in C.fluminea. This system has made of one
main tank (capacity of 100 liter) and four tanks (capacity of 2/5 liter) for standing of clams (one of them as control) and one tank
for returning of water. In this research, three different treatments of Nitrogen (80, 100 and 200 ppm) and Phosphorus (20, 40 and
60 ppm) were separately examined. Three times sampled at all treatments . Our observations showed that filtration rates of pollutant
material from the beginning of experiments were decreased at tree treatments and also filtration rates of clams increased
by increasing of Nitrogen concentrations so that filtration rates have reached from 6/53 in low treatment to 7/15 in medium
treatment and 8/07 ml.min -1 .gr -1 (AFDW) in high treatment. Filtration rates of Phosphorus were also increased as filtration rates
have reached from 5/61 in low treatment to 6/5 in medium treatment and 7/58 ml.min -1 .gr -1 (AFDW) in high treatment. Comparison
between filtration rate of Nitrogen and Phosphorus showed that clam’s efficiency in absorbing of Nitrogen compounds
is more than Phosphorus compositions.

EVALUATION OF THE PREBIOTIC GROBIOTIC ® - A IN THE DIET OF JUVENILE
COMMON GOLDFISH (Carassius auratus) IN THE PRESENCE OR ABSENCE OF NATURAL
PRODUCTIVITY
L. Christine Savolainen*, Alejandro Buentello, Gary Burr and Delbert M. Gatlin
Department of Wildlife and Fisheries Sciences
Texas A&M University System
College Station, Texas 77843-2258 USA
christinesavolainen@tamu.edu
Gastrointestinal (GI) tract microbiota have been reported to play a significant role in the health and nutrition of humans and
terrestrial animals. However, research in this area has been lacking in fish species produced in aquaculture. Because some
prebiotics recently have been shown to increase immune responses and disease resistance of certain fish species, the current
study was conducted to evaluate the commercial prebiotic GroBiotic ® -A with juvenile goldfish (Carassius auratus). Goldfish
are a prominent bait and ornamental species in the U.S. and are subjected to various stressors and potential diseases during their
aquacultural production.
The current study consisted of two, 12-week feeding trials in which juvenile goldfish were fed practical diets of the same
composition either unsupplemented or supplemented with 1% GroBiotic ® -A by weight. GroBiotic ® -A is a mixture of partially
autolyzed brewers yeast, dairy components and fermentation products, In each feeding trial, juvenile fish were sorted by size
and stocked into 12 units within each of two culture systems. One culture system without natural productivity consisted of 38-L
aquaria supplied with recirculated well water maintained at 26ºC ± 1ºC throughout the trial. The system which provided natural
productivity consisted of 19-L plastic buckets contained within a 1200-L fiberglass tank which received a continuous supply
of pond water. A total of 20 fish initially averaging 1.38 g were stocked into the aquarium system and 10 fish averaging 1.74
g were stocked into the bucket system. Each of the experimental diets was randomly assigned to six units within each culture
system and fed to fish at a fixed percentage of body weight divided into two feedings per day for a total of 12 weeks.
Significantly (p

30
SALINITY TOLERANCE OF SUNRAY VENUS CLAM Macrocallista nimbosa NURSERY SEED
John Scarpa*, Michelle Harangody and Leslie N. Sturmer
Harbor Branch Oceanographic Institute at Florida Atlantic University
Center for Aquaculture and Stock Enhancement
5600 U.S. Hwy 1 North
Ft. Pierce, FL 34946 USA
jscarpa1@hboi.fau.edu
The sunray venus clam Macrocallista nimbosa was commercially fished in Florida during the 1970s. Although natural growth
rates were estimated to be high, its patchy distribution limited commercial exploitation. The sunray venus clam is now being
evaluated as a potential new aquaculture species to diversify the hard clam culture industry in Florida. Therefore, basic physical
and biological parameters for optimum culture environments need to be determined. In this study, the salinity tolerance of
nursery seed was examined.
Three separate families (55, 61 & 85) of sunray nursery seed clams were utilized as replicates and exposed to salinities of 10,
20, 30 or 40 ppt in triplicate 4-L beakers. Twelve individuals from each family (avg initial whole wt = 19±3 mg, avg initial
length = 4.7±0.3 mm) were utilized in each beaker (n=36 clams/beaker). Individual clams were placed in separate openings of
a plastic grid (1.2 x 1.2 cm openings) with screen (1.0 mm openings) on the bottom. The trays were suspended approximately
8 cm from the bottom in 3 L of treatment water, which was gently aerated. Clams were fed twice per day with the microalgae
Isochrysis sp. at a density of 100,000 cells/mL; water was changed completely every other day. Clams were examined daily
for mortality and weighed weekly for three weeks.
All clams in the 10 ppt treatment died by the end of week 1. After three weeks, survival at 30 ppt (81%) was significantly
greater than at 40 ppt (55%), but not from 20 ppt (69%). Survival was not significantly different between 40 ppt and 20 ppt.
Interestingly, family 85 exhibited significantly lower survival (50%) as compared to family 61 (74%) and 55 (81%), which
were not different from each other.
Growth (% whole wt change) for surviving clams was significantly affected by salinity. After three weeks, clams at 30 ppt
grew significantly more (270%) than those at 40 (208%), which grew significantly more that those at 20 ppt (125%). Family 85
exhibited significantly lower growth (171%) as compared to family 61 (225%), but was not different from 55 (207%). Family
61 and 55 were not different from each other.
These results indicate that the optimum salinity for growth and survival of sunray venus clam nursery seed is 30 ppt. Additionally,
the differences found between families imply potential for increasing growth and survival through selective breeding. This
research was supported by Florida Sea Grant (Projects R/LR-A-44 and R/LR-A-45).

GENETIC DIVERSITY OF CULTURED AND WILD POPULATIONS OF THE FRESHWATER
PRAWN Macrobrachium rosenbergii BASED ON MICROSATELLITE ANALYSIS
Kyle J. Schneider*, James H. Tidwell, Boris Gomelsky, Kirk W. Pomper and Geoffrey C. Waldbieser
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
kyle.schneider@kysu.edu
The freshwater prawn Macrobrachium rosenbergii is a popular aquaculture species worldwide. Historically, cultivation began
in Malaysia during the early 1960s. Shortly after, prawns were introduced to the Anuenue Fisheries Research Center (AFRC)
in Hawaii where mass rearing techniques for commercial scale production of post larvae were developed. It is documented that
only 36 individuals were introduced to the AFRC from Malaysia. During the 1970s, cultivation of freshwater prawn expanded
into many areas outside the native range of this species. Prawns were introduced from Hawaii to many locations including
North and South America, Europe, Africa and Asia. Based on these records, it may be suggested that due to the restricted
number of initial animals (founder effect) that many cultivated stocks likely exhibit low levels of genetic diversity. The goal
of this research was to investigate the genetic diversity of freshwater prawns from different populations based on microsatellite
analysis with a focus on North American populations.
A total of nine prawn samples were collected from various cultured stocks and wild populations. Genetic variability was assessed
at six microsatellite loci. Amplified products were resolved via capillary electrophoresis on a ABI 3730xl DNA analyzer
platform. The results of this analysis revealed that many of the cultured stocks evaluated exhibit a reduction in genetic diversity.
The average number of alleles per locus ranged from 3.83 to 23.00 and average expected heterozygosity (Nei 1978) ranged
from 0.584 to 0.937 with the Myanmar population (wild) exhibiting the greatest diversity and the Israel stock (cultured) exhibiting
the lowest diversity (Table 1). Based on these results it is apparent that the compound founder effect of cultured stock
establishment along with generations of breeding without introduction of new individuals has negatively impacted diversity.
Table 1. Summary of sample size (N), average number of alleles per locus (A), average
observed herterozygosity (Ho), average expected heterozygosity (He) and inbreeding
coefficient (Fis) for nine populations of M. rosenbergii across six microsatellite loci.
Population Hawaii Hawaii India India Israel Kentucky Mississippi Myanmar Texas
N 49 1 60 2 45 1 50 2 50 60 49 60 50
A 10.67 11.00 18.17 5.17 3.83 6.50 6.17 23.00 5.67
Ho 0.790 0.811 0.860 0.659 0.723 0.625 0.653 0.906 0.697
He 0.847 0.860 0.907 0.667 0.584 0.719 0.730 0.937 0.681
Fis
0.068 0.057 0.052 0.011
-
0.240 0.131 0.106 0.033
-
0.023
30

310
GEOSMIN PRODUCTION BY Nocardia SPP. and Streptomyces SPP. FROM RECIRCULATING
AQUACULTURE SYSTEMS (RAS) FOR RAINBOW TROUT Onchorhynchus mykiss CULTURE
Kevin K. Schrader and Steven T. Summerfelt
USDA, ARS, NPURU
Thad Cochran Research Center
University, MS 38677-8048 USA
Pre-harvest “off-flavor” in aquaculture products results in large economic losses to producers due to delayed harvest. “Earthy”
and “musty” are the most common off-flavors reported in fish raised in ponds, and these off-flavors have also been reported in
fish raised in recirculating aquaculture systems (RAS). The presence of the microbial metabolites geosmin and 2-methylisoborneol
(MIB) in the flesh of fish are responsible for earthy and musty off-flavors, respectively. Although cyanobacteria are
responsible for most earthy-musty problems in pond-cultured fish, the microbial sources of geosmin and MIB in RAS are still
being investigated.
In this study, we tested for geosmin and MIB off-flavor compounds within six replicated recirculating systems producing
rainbow trout (Onchorhynchus mykiss). Water, biosolids/biofilm, and fish samples were collected twice during an ozonation
study: 1) when fish were at maximum feed levels and densities (69 kg/m 3 ); and 2) approximately two months later and just
prior to termination of the study. Each RAS contained a fluidized-sand biofilter, a forced-ventilated cascade aeration column,
a low head oxygenation (LHO) unit where pure oxygen feed gas was absorbed, a LHO sump, a single 5.3 m 3 culture tank, a
microscreen drum filter, a particle trap, a pump sump, and a heat exchanger. Approximately 99.74% of the water flow was
recirculated in each system, which provided mean system hydraulic retention time of approximately 6.7 days. The oxygen feed
gas transferred into the recirculating water was ozonated in three of the systems and not ozonated in three systems. A constant
24-h photoperiod was provided and all systems were fed equal portions once every 2 hours. Solid phase microextraction-gas
chromatography-mass spectrometry (SPME-GC-MS) was used to determine the presence and levels of geosmin and MIB in
water, biosolids, and trout fillet samples. In addition, we investigated the potential microbial sources of earthy off-flavor in
the flesh of RAS-cultured rainbow trout. Water and biosolids samples were used to inoculate media that is utilized to culture
microorganisms capable of producing geosmin and/or MIB (e.g., actinomycetes, cyanobacteria, fungi, and myxobacteria).
Microbial isolates identified as producing an earthy-musty odor were subjected to SPME-GC-MS analysis to verify geosmin
and/or MIB production.
Geosmin levels were higher and more prevalent than MIB in water, biosolids, and trout fillets from the six RAS, with highest
geosmin levels in biosolids samples compared to water samples. Four species of earthy-odor producing actinomycetes,
Nocardia cummidelens, Nocardia fluminea, Streptomyces luridiscabiei, and Streptomyces cf. albidoflavus, were isolated from
biosolids contained within the RAS drum filters and heat exchangers, and these isolates were subsequently confirmed to be
geosmin producers. No other geosmin-producing microorganisms were isolated during our study. We concluded that these
geosmin-producing isolates were the likely source of geosmin in the RAS water and contributors to the earthy off-flavor in the
trout.

PROGRAM EVALUATION
Michael H. Schwarz
Virginia Seafood Agricultural Research and Extension Center
102 South King Street
Hampton, VA 23669 USA
mschwarz@vt.edu
SKYPE: michaelhschwarz
The final component of any extension program is program evaluation and or accountability. However, program evaluation is a
continuous process and to be effective must be taken into account during all phases of program development. Program evaluation
is a plan for implementation of procedures to measure a program’s success or impact (summative evaluation), to improve
the program (formative evaluation), and/or report results to program stakeholders. Program evaluation includes the following:
1) Identification if the evaluation is to be formative or summative; 2) identification and design of appropriate survey instrument
data collection techniques that will be utilized; 3) determination of survey instrument validation and reliability; 4) collection of
evaluation data; 5) survey data analysis; and 6) reporting results. This presentation will provide an overview of these evaluation
components, providing additional discussion on potential sources of survey error.
311

312
HEAT SHOCK PROTEIN REGULATION IN RAINBOW TROUT Oncorhynchus mykiss IS
ALTERED BY DIETARY SOYBEAN MEAL INCLUSION AND ANTI-PHOSPHOLIPASE A 2
ANTIBODY
Wendy M. Sealey * , Frederic T. Barrows, Charlie E. Smith, Jurij M. Wacyk, Brian C. Donahower,
Madison S. Powell, Ronald W. Hardy and Eric A. Shelden
Hagerman Fish Culture Experiment Station
University of Idaho
3059F National Fish Hatchery Road
Hagerman, ID 83332 USA
wsealey@uidaho.edu
Heat shock proteins (HSPs) are considered to be a cell survival factor and have been shown to improve cell function in multiple
models of cell injury including soybean meal-induced enteritis in salmonids. Recently, phospholipase A 2 , an enzyme that catalyzes
the hydrolysis of fatty acids to generate arachidonic acid, was shown to regulate HSPs. Thus alteration of phospholipase
A 2 activity may provide an opportunity to alter heat shock protein expression and reduce soybean meal-induced enteritis in
rainbow trout. Therefore, the aim of this study was to examine the effect of a commercially available anti-phospholipase A 2
supplement (Big Fish) on performance of rainbow trout fed high levels of soybean meal (SBM).
Rainbow trout were fed a practical-type control diet formulated to contain 45% protein and 15% fat with 0, 17.5 or 35% SBM
with and without the commercially available Big Fish (Aova Technology) supplement at an inclusion level of 0.3%. Diets were
fed to three replicate tanks of fish per treatment (20 fish/tank, House Creek strain) for 9 weeks. Trout were reared in 80 L tanks
supplied with 4 L/min of 14.8°C flow-through spring water. Following the trial, fish (n=3) were randomly selected from each
tank for whole body proximate analyses and fish performance indices. Total RNA was isolated from liver and intestinal tissues
(n=3) to detect alterations in HSP27, HSP70 and HSP90 gene expression by real time quantitative PCR; protein expression
was examined by Western blot. Histopathological examination was performed on the kidney, liver and distal intestine of three
randomly selected fish from each tank.
Decreased growth was observed for fish fed 35% SBM regardless of Big Fish supplementation. Fish fed 35% SBM had significantly
higher feed conversion ratios than fish fed 0 or 17.5% SBM. HSP27 and HSP90 mRNA expression was elevated in
fish fed 35% SBM. A significant interaction was observed for HSP90 mRNA expression in that fish fed 35% SBM with Big
Fish had lower HSP90 mRNA expression that fish without supplement. Histology of the distal intestine revealed significant
pathology in fish fed 35% SBM diets (with and without Big Fish). These data indicate feeding high levels of SBM alters the
expression and regulation of various HSP mRNA of rainbow trout in a HSP specific manner. Additionally, dietary inclusion of
Big Fish had only minimal effects on the pathology associated with high levels of SBM in rainbow trout.

SENSORY ANALYSIS OF RAINBOW TROUT Oncorhynchus mykiss FED PUFA-ENRICHED
BLACK SOLDIER FLY PREPUPAE
Wendy M. Sealey * , T. Gibson Gaylord, Frederic T. Barrows, Jeffery K. Tomberlin, Mark A. McGuire, Carolyn Ross
and Sophie St-Hilaire
Hagerman Fish Culture Experiment Station
University of Idaho
3059F National Fish Hatchery Road
Hagerman, ID 83332 USA
wsealey@uidaho.edu
Limited work has been done on the use of insects for fishmeal and fish oil replacement in rainbow trout diets, despite the fact
that insects make up part of their natural diet. A previous study in our laboratory indicated that black soldier fly prepupae could
be used to replace a portion of the fishmeal component of a practical grow-out diet without compromising growth. However, in
that study, sensory analysis of fillets was not performed. Therefore, the objective of this study was to determine the effects of
dietary inclusion of black soldier fly prepupae on rainbow trout fillet quality.
Black soldier fly prepupae were reared on dairy manure and enriched by supplementing fish offal to the growth medium during
the last month of culture. For the fish feeding trial, a practical-type trout diet was formulated to contain 40% protein with roughly
half of the protein derived from fish meal and the remaining from soybean meal, corn gluten meal and wheat meal. Four test
diets were developed by substituting 25% and 50% of the fishmeal component of the control diet with normal (BSF) or enriched
(EBSF) black soldier fly prepupae on a amino acid equivalents basis. Dietary fat content was adjusted to approximately 15%
lipid using fish oil and poultry fat to provide required levels of essential fatty acids and maintain approximately equivalent fatty
acid ratios between the treatments. Diets were fed to three replicate tanks of fish per treatment (15 fish/tank, mixed-sex House
Creek strain) for 8 weeks. Trout were reared in 140 L tanks supplied with 6 L/min of constant temperature (14.8°C) flowthrough
spring water. At the end of the feeding trial, three fish per tank were sampled for determination of hepatosomatic index,
intraperitoneal fat ratio and muscle ratio. The muscle portion obtained was subsequently utilized for determination of proximate
and fatty acid composition. Fish remaining after sampling were pooled by tank, euthanized and immediately transported on ice
to a commercial fish processing facility. At the processing facility, fish were individually hand-filleted as gourmet fillets with
ribs and pin bones removed. Following filleting, samples were pooled by tank and stored at -20C until shipment (approximately
1 week) to Washington State University for sensory evaluation.
Growth of fish fed the EBSF diets was not significantly different from those fish fed the fish meal-based control diet while
growth of fish fed the BSF diets was reduced as compared to the control diet. Muscle ratio was not altered by diet but control
fish had significantly higher IPF than those fish fed the EBSF or BSF diets. A group of 30 untrained panelists did not detect a
significant difference in a blind comparison of fish fed the fish meal containing control diet as compared to fish fed the EBSF
or BSF diets. These data suggest that EBSF can be used to replace up to 50% of fishmeal portion of a practical trout diet for 8
weeks without significantly affecting growth or sensory quality of rainbow trout fillets.
313

314
EVALUATION OF THE ABILITY OF CHICKEN 42 POULTRY PRODUCT, CHICKEN 70%
POULTRY BY-PRODUCT BLEND, AND CHICKEN AND EGG CONCENTRATE (102190) TO
REPLACE FISH MEAL IN RAINBOW TROUT Oncorhynchus mykiss STARTER DIETS
Wendy M. Sealey * , Frederic T. Barrows, Qing Pan, David A. J. Stone and Ronald W. Hardy
Hagerman Fish Culture Experiment Station
University of Idaho
3059F National Fish Hatchery Road
Hagerman, ID 83332 USA
wsealey@uidaho.edu
Poultry meal products have been successfully used to replace fish meal in diets for rainbow trout. However, the potential for
these products to replace fish meal depends on consistency of their digestible nutritional composition. Therefore, the aim of the
current study was to assess the ability of three different commercially-available branded poultry products to maintain growth
and disease resistance of juvenile rainbow trout Oncorhynchus mykiss when substituted for 100% of the fishmeal component
of a typical starter diet.
A practical-type fish meal-based starter diet was formulated to meet or exceed the nutrient requirements of juvenile rainbow
trout and to contain 48% crude protein and 18% crude lipid. Test diets were formulated by substituting 100% of the fishmeal
with Chicken 42 poultry product, Chicken 70% protein poultry by-product blend (Chicken 70), and Chicken and Egg Concentrate
(102190). Rainbow trout (College of Southern Idaho, House Creek strain) were used in this study. Fertilized eggs were
obtained, hatched and fry grown to approximately 0.5 g. At stocking, rainbow trout were counted into groups (50 fish) with six
replicate tanks of fish for each diet. Trout were reared in 140-L tanks supplied with 4 L/min of constant temperature (14.8°C)
flow-through spring water and fed their respective diets four times daily for 8 weeks. Following the feeding trial, fish (n=3)
were randomly selected from each tank for whole body proximate analyses and fish performance indices. To examine disease
resistance, a sub sample of fish from each tank was challenged by subcutaneous injection with Flavobacterium psychrophilum.
Fish were monitored for mortality and fed their respective diets for 28 d following challenge.
All of the poultry product-based diets supported growth, nutrient retention and feed conversion ratios of rainbow trout fry equal
to or greater than those observed for fry fed fish meal-based diets. Fish fed Chicken 42 had significantly greater growth than
all other diets; fish fed Chicken 70 had similar growth to those fed fish meal and both had reduced growth as compared to those
fish fed Chicken and Egg Concentrate or Chicken 42. Protein and energy retention efficiencies reflected those trends observed
for growth. Feed conversion ratios were acceptable for all diets and ranged from 0.83 to 1.05. In contrast, no significant effect
of diet was observed following experimental challenge with Flavobacterium psychrophilum. These data suggest that the
examined poultry products can be used in place of fish meal in starter diets for rainbow trout fry without detrimentally affecting
growth and disease resistance.

POULTRY BY-PRODUCT MEAL- BASED PELLETED FEEDS FOR THE CULTURE OF
HUMPBACK GROUPER Cromileptes altivelis
Rossita Shapawi*, Saleem Mustafa and Ng Wing Keong
Borneo Marine Research Institute
Universiti Malaysia Sabah
Locked Bag 2073
88999 Kota Kinabalu Sabah, Malaysia
rossita@ums.edu.my
Two isoproteic and isolipidic feeds (U-MS/SM1 and U-MS/SM2) were formulated with a blend of alternative ingredients in
the place of marine fish meal and fish oil, and fed to humpback grouper (Cromileptes altivelis), a tropical marine carnivorous
fish species. Both prototype feeds were formulated with poultry by-product meal (PBM) to replace fish meal at different inclusion
levels. U-MS/SM1 contained less dietary PBM level than U-MS/SM2. Added dietary lipid was in the form of crude palm
oil (CPO) and fish oil in U-MS/SM1, and fish oil in U-MS/SM2.Growth performance, feed utilization efficiency, survival and
cost-benefits of humpback grouper fed these diets were compared with those fed ComDiet1 (local commercial marine fish
feed), ComDiet2 (imported high-fat commercial marine fish feed) and trash fish (Sardinella spp.). The performance of U-
MS/SM1 and U-MS/SM2 feeds was similar to or better than the imported marine fish feed, ComDiet2. Both U-MS/SM1 and
U-MS/SM2 feeds, give better growth compared to fish fed ComDiet1 or the trash fish. In case of fish fed U-MS/SM2, growth
was significantly higher than those given U-MS/SM1 and ComDiet2, three times higher than the growth of fish fed ComDiet1,
and double the growth of fish fed the trash fish (Table 1). In conclusion, diets U-MS/SM1 and U-MS/SM2 were able to reduce
the feed cost per kg of humpback grouper produced (Table 2).
31

31
A COMPARISON OF TWO ALUMINUM BASED COAGULANTS FOR PHOSPHORUS
PRECIPITATION AND SUSPENDED SOLIDS REMOVAL USING INCLINED BELT FILTER
TECHNOLOGY TO DEWATER RECIRCULATING AQUACULTURE EFFLUENT
Mark J. Sharrer, Kata Rishel and Steven T. Summerfelt
Conservation Fund Freshwater Institute
1098 Turner Road
Shepherdstown, WV 25443 USA
m.sharrer@freshwaterinstitute.org
Aquaculture production methods typically produce a high volume fish farm effluent that contains low concentrations of organic
material and nutrients that can result in adverse affects on a receiving water body. Effective removal of particulate and dissolved
fractions of these pollutants is necessary to adhere to state and federal regulatory guidelines and for proper overall fish farm
management. Efficient collection of particulate waste products is evident in recirculating aquaculture systems (RAS) technology.
Backwashing and flushing of mechanical filters and gravity settling units results in a small volume of waste concentrated
effluent relative to the entire recirculating volume. When further wastewater treatment is desired, this reduction in volume
increases treatment efficiency and reduces cost of on-site treatment options.
Inclined belt filter technology for wastewater treatment utilizes polymer / coagulant amendments to aggregate solids and precipitate
dissolved phosphorus, separating these constituents from final discharge. The purpose of this research was to flocculate
fish culture wastewater with a polymer and compare the phosphorus adsorption capacity and optimal dosing requirements of
either aluminum chloride (AlCl 3 ) or aluminum sulfate (Alum) prior to inclined belt filter dewatering. A series of bench-scale jar
tests applying each coagulant (without polymer) determined the most effective dose based upon total suspended solids (TSS)
and dissolved reactive (DRP) phosphorus concentrations in jar supernatant. A subsequent series of jar tests determined the
optimal combination of polymer dose and previously determined coagulant (AlCl 3 and Alum) doses.
Based upon the jar test analyses, four flocculant/coagulant doses were compared utilizing an inclined belt filter operated at the
Conservation Fund Freshwater Institute for solids dewatering from commercial scale recirculating rainbow trout (Oncorhynchus
mykiss) production. Chemical amendment treatments applied were 1) 10 mg/L polymer (control), 2) 10 mg/L polymer +
290 mg/L AlCl 3 , 3) 10 mg/L polymer + 50 mg/L Alum, and 4) 10 mg/L polymer + 146 mg/L AlCl 3 (stoichiometric equivalent
of 50 mg/L Alum). Results indicate that filtrate TSS removal was >98.8 when applying any coagulant in combination with the
polymer. Further, DRP removal was most efficient applying AlCl 3 at 290 mg/L (95.9%) in combination with the polymer.
Table 1. Percent removal of key water quality parameters at each treatment

SOME COMMON DESIGN AND STATISTICAL ANALYSIS PROBLEMS APPEARING IN
THE AQUACULTURE LITERATURE
Karl D. Shearer
The Aquaculture Protein Centre
Department of Animal and Aquacultural Sciences
The Norwegian University of Life Sciences
1432 Aas, Norway
Karl.shearer@umb.no
Statistical analysis is an integral part of aquaculture research. The widespread availability of desktop computers and relatively
inexpensive statistical software has allowed researchers to deign and evaluate increasingly more complex experiments. There
is however, widespread misuse of these tools. It is essential that researchers have sufficient statistical literacy to properly design
experiments, choose an appropriate statistical test and verify that the underlying assumptions of the test have been met.
This applies not only to their own research, but also to their ability to evaluate published literature and to referee manuscripts
submitted for publication. Examples, from the aquaculture literature, of some common design and statistical analysis mistakes
will be presented along with some suggestion that can help avoid some of these problems. Examples are taken from the most
common types of experiments; nutrient requirements, dietary ingredient evaluation and fish meal and oil replacement, proximate
composition determination, protein/energy balance, dietary affects on physiology, fish health evaluation and product quality
determination. This presentation will concentrate on conceptual rather than the computational aspects of these problems.
Resources that can be used to aid in evaluation of the statistical quality of aquaculture literature are presented.
31

31
AQUACULTURE SUSTAINABILITY, ARE OFFSHORE REARING SYSTEMS AND PLANT
BASED FEEDS THE ANSWER?
Karl Shearer
Aquaculture Protein Centre, The Norwegian University of Life Sciences
P. O. Box 5003
NO-1432 Ås, Norway
karl.shearer@umb.no
In 2006 aquaculture provided 43% of the fish used for human consumption, and production is currently growing at approximately
9% per year. Expectations are that this growth will continue at a similar rate for the foreseeable future. Although current
aquafeed production, about 15 million t, is a small portion of the animal feed industry (currently over 600 million t) it will
likely become a larger portion in the future. Estimates of demand for aquafeed in 2030 are as high as 80 million t. A number of
concerns about the impact of aquaculture on the environment have been raised by NGOs and two items of specific concern to
those involved with aquafeed are (1) the feeding of fish to fish and (2) the pollution caused by rearing fish in open systems. In
addition, the question of sustainability is of concern to producers, retailers, consumers and the general public.
The aquafeed industry, along with a majority of researchers in this area claim that the industry is working towards sustainability.
Using the 1987 UN Brundtland Report definition of sustainability “meeting the needs of the present without compromising
the ability of future generations to meet their own needs” I will present an examination of this claim. Examining the industry
response to concerns (1) and (2) above and looking at the source and ultimate fate of the nutrients contained in aquafeeds
suggests that we are proceeding down the wrong path to achieve sustainability.
Aquaculture should be viewed as one of many components of a global food production system that share a single nutrient pool.
Sustainability is maintained when nutrients can be moved between components and re-used, but conversely when nutrients are
lost or so diluted as to not be reclaimable (e.g. if they are discharged into the world’s oceans), then sustainability is not maintained.
This situation will be illustrated using phosphorus as an example, as phosphorus may be the first limiting nutrient for
industrial food production in the very near future. Processes that can improve sustainability are available but require additional
research for them to be optimized and for the production systems to be economically viable.

PADDLEFISH SYMPOSIUM DEDICATION TO JAMES KAHRS
William Shelton and Steven Mims
Zoology Department
University of Oklahoma
Norman, OK USA
wshelton@ou.edu
James “Jim” W. Kahrs was a dedicated fish farmer, a congenial person, and a good friend. He established the Osage Catfisheries
in 1953, and he operated until his retirement in 1996. It became a family business and continues today under the guidance of
his sons. Jim was recognized by the American Fisheries Society, Fish Culture Section with his nomination induction into the
National Fish Culture Hall of Fame, Spearfish, SD in 1999. Jim was also active in many other professional organizations such
as the National Aquaculture Association, Catfish Farmers of America, and the World Aquaculture Society. Jim was active in
interacting with State, Federal and International fishery workers. The paddlefish has been a popular sportfish and much of the
early developments in artificial propagation were developed to facilitate stocking programs. Jim became interested in paddlefish
for its potential as a commercial species. This activity led him into international trade, promoting paddlefish as a foodfish.
We remember Jim who died in September 2006 by dedicating this paddlefish symposium convened at the Aquaculture America
2009 meeting in Seattle, WA.
31

320
MONOSEX PRODUCTION OF PADDLEFISH - REVISITED
William L. Shelton*, Steven D. Mims, Boris Gomelsky and Richard J. Onders
Zoology Department
University of Oklahoma
Norman, OK 73019 USA
wshelton@ou.edu
Artificial propagation of the North American paddlefish (Polyodon spathula) has been used primarily for maintenance and
restoration of fisheries. However, interest in their culture as a foodfish has grown internationally. Paddlefish have high
quality meat and darkly pigmented eggs. All-female culture is of particular interest relative to caviar production. Development
of a breeding program to produce only female progeny would offer an optimal culture system. A research program at
Kentucky State University has focused on this goal over the past two decades. Components include phenotypic and genetic
manipulations, but the genetic mechanism of sex determination is central to program. Induction of gynogenesis is a primary
component. Treatment optimization for ploidy manipulation can be achieved, and advances have been made in hormonally
induced sex reversal. Relative to sex determination, the original thesis for a breeding program proposed a homogametic (XX) female
system; however, this hypothesis was based on a small sample size of immature gynogenotes. Recent evaluation of a population
of sexually mature gynogenotes has provided evidence that the genetic basis of sex determination in paddlefish is probably
one with heterogametic (WZ) females.
The harvest of 8-year-old gynogenotes provided unequivocal evidence that the premise of an XX-system was incorrect. A
heterogametic female system is probable, although the observed sex ratio departed from the theoretical ratio of 1:1 which would
be expected for gynogenotes of a WZ-female. Fifty-eight percent (105/180) of the fish were females with mature ova, while
nineteen percent (35/180) were mature males. The GSI for the presumptive WW-females was equivalent to females with the
normal genotype (WZ). If the WW-genotype of gynogenote females is verified, alternate management pathways might permit
the development of an alternate breeding program for monosex female paddlefish.

FORECASTING POND BANK PRICES OF CATFISH IN U.S.A: AN APPLICATION OF
VECTOR AUTOREGRESSION MODEL
Kehar Singh and Madan Mohan Dey
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
1200 N University Drive, Mail slot 4912
Pine Bluff, AR 71601 USA
ksingh@uaex.edu
The catfish farmers in the USA sell more than 95% of their produce to processors, and the prices are generally determined on
spot. Review of existing literature show that many farmers have weak bargaining position in negotiating price with processor.
The knowledge of likely trends in future pond bank prices (PBP) of catfish will improve farmers’ bargaining power, and will
also help them to adjust supply to maximize their returns. This paper has used vector autoregessive model (VAR) to forecast
monthly pond bank price for catfish in the USA.
The VAR model is a time series modeling technique, which produce forecasts by extrapolating the historical behaviour of
variable(s) of interest. The model can capture the interdependencies between multiple series. It is a ‘theory-free’ method to
estimate economic relationships, and is an alternative to the structural model that must satify a number of theoritical ‘restrictions’.
We have postulated that PBP is the function of its past values, past values of processors’ demand, past values of catfish,
basa and tra imports, and past values of fuel prices.
The study has utilized two sets of time series data: i) monthly data from January 1990 to September 2008, and ii) quarterly data
from first quarter of 1990 to third quarter of 2008. The data for t th month generally become available during the third week of
t+1 th month. As the VAR model generates forecast for one period, the forecast for t+1 th month can be made available during
the last week of t+1 th month. Therefore, we have used different alternative procedures to make forecast for t+1 th , t+2 th and t+3 th
months by i) combining monthly and quarterly forecasts, ii) adjusting them for seasonal and recent past forecast errors, and iii)
using the forecasts of explanatory variables.
The paper compares the results from alternative approaches and makes suggestions for the use of the results under different scenarios.
We have used ‘Pseudo Out-of-Sample Forecasting’ method to i) simulate the real-time performance, and ii) test predictive
power by estimating ‘Root Mean Square Forecasting Error. The statistical tests confirm the stability of the coefficients and
the non-significant auto-correlations of residuals in the alternative models. Results show that it is possible to forecast monthly
PBP for catfish within reasonable confidence bounds.
321

322
MANAGEMENT OF GENETIC DIVERSITY IN PADDLEFISH Polyodon spathula
POPULATIONS AND PROPAGATION
Brian L. Sloss*, Ryan P. Franckowiak, Ed Heist and Robert Klumb
Wisconsin Cooperative Fishery Research Unit
U.S. Geological Survey
College of Natural Resources
University of Wisconsin-Stevens Point
Stevens Point, WI 54481 USA
bsloss@uwsp.edu
The genetic integrity of native populations of paddlefish (Polyodon spathula) can be threatened by propagation through two
primary avenues: the use of genetically divergent source populations as broodstock and the propagation of genetically depauperate
individuals due to insufficient broodstock numbers or crossing/rearing strategies. The basic information necessary to
conduct genetically-sound propagation of paddlefish is to understand the levels and distribution of genetic diversity throughout
the paddlefish’s range and the genetic impact of different crossing and rearing strategies to allow for predictive management of
the genetic diversity of propagated paddlefish. Studies using mitochondrial DNA and microsatellite DNA, although not sufficient
to recognize cohesive stocks of paddlefish, have shown significant genetic structure throughout the paddlefish’s native
range suggesting multiple genetic management units should be enforced.
An example of a genetic assessment prior to wild propagation in one of these units is a series of studies conducted in association
with the upper Missouri River paddlefish propagation. The objectives were twofold: 1) to determine if significant differences
existed among broodsources (Gavins Point tailrace, Niobrara River/Missouri River confluence, and Lake Francis Case/White
River) used in the USFWS’s Missouri River paddlefish propagation conducted at the Gavins Point Dam National Fish Hatchery,
and 2) to determine if male reproductive success significantly deviated from random expectation when a mixed-milt design
(traditional) versus a split-egg lot design (experimental) were used by testing larval fish using genetic paternity analysis. The
number of males used in the three replicate mixed-milt crosses ranged from 4 to 9 whereas the split-lots used 5 males each. All
potential broodsources were genetically similar based on tests of genic differentiation and none deviated from the paddlefish
harvested in a snag fishery located near Gavins Point. Significant male contributions (χ 2 p max -value < 0.0001) were observed
among all traditional crosses with male contribution ranging from 86% to

C-TERMINAL MODIFICATION MEDIATES BIOLOGICAL ACTIVITY OF GHRELIN IN
CHANNEL CATFISH
Brian C. Small
USDA/ARS Catfish Genetics Research Unit
Thad Cochran National Warmwater Aquaculture Center
P.O. Box 38
Stoneville, MS 38776 USA
Brian.Small@ars.usda.gov
Ghrelin is a multifunctional peptide hormone secreted by the stomach and originally identified as the endogenous ligand for
the growth hormone secretagogue receptor (GHS-R). In vertebrates, ghrelin functions primarily as a growth hormone (GH)releasing
hormone and an appetite-stimulatory (orexigenic) hormone. Ghrelin has been isolated and partially characterized in
several mammalian and non-mammalian species, including fish. Ghrelin mature peptide has been identified in goldfish, eel,
Mozambique tilapia, Nile tilapia, rainbow trout, seabream, European sea bass, zebrafish, and channel catfish. Fish ghrelins are
typically 19-23 amino acids and exhibit GH-, prolactin (PRL)-, or luteinizing hormone (LH)-releasing activity.
Channel catfish ghrelin peptide and cDNA encoding precursor protein were isolated from the stomach of a channel catfish. In
addition to the amidated 22-amino acid ghrelin peptide, a Gly-extended, non-amidated 23-amino acid ghrelin (ghrelin-gly) was
isolated. Characterization of the two forms of channel catfish ghrelin indicates differential actions on GH-releasing activity,
receptor regulation, and tissue specificity. These results demonstrate C-terminal modification mediates biological activity in
channel catfish. Such an effect of C-terminal modification has not been previously described for any other vertebrate ghrelin.
323

324
THE CASE FOR MARINE FINFISH AQUACULTURE IN ALASKA
Scott Smiley
University of Alaska Fish Tech Center
118 Trident Way
Kodiak AK 99615 USA
smiley@sfos.uaf.edu
NOAA Fisheries presented their plan for the development of offshore finfish aquaculture in Alaska. They chose Pacific halibut
(Hippoglossus stenolepis) and sablefish or black cod (Anoplopoma fimbria) as models. These species are currently harvested
in Alaska under IFQ permits that are traded and have a considerable value that fluctuates in the market place. Unfortunately for
NOAA Fisheries, many Alaskans had purchased IFQs, some using their life savings. The notion that the Government would
assist in building an off shore aquaculture business to raise halibut and black cod was perceived as a financial threat to these
fishermen. Shortly afterwards the Alaska State Legislature outlawed marine finfish aquaculture in Alaskan state waters, out to
3 miles. Subsequently the Alaskan Congressional delegation made it clear that participation in NOAA fisheries plan to develop
a marine aquaculture industry in Alaska’s US federal waters (3 to 200 miles off shore) would require permission of the state.
Aside from the perceived threat to their financial well-being, Alaskan harvesters have seen the consequences of competition
with a marine aquaculture system in salmon. Once Alaska was world leader in the salmon industry, but over the past 25 years,
competition from pen reared salmon has changed the market opportunities for Alaska’s wild fish. Atlantic (Salmo salar) and
coho salmon (Oncorhynchus kisutch) have become the price leaders in many US retail outlets and farmed salmon define high
quality. This has alienated many Alaskan salmon fishing families who do not entertain the idea of marine finfish aquaculture
in Alaska.
Yet, Alaska has allowed and practiced salmon aquaculture though its hatchery system for more than 30 years. Employing what
is termed Ocean Ranching, these hatcheries account for roughly a third of all the salmon harvested in Alaska. In 2007, more
than 212 million salmon were harvested and more than 77 million, or 36% were derived from Private-Non-Profit (PNP), state
and federal hatcheries, which took from the common property fishery 19 million fish for their cost recovery. The hatcheries collect
gametes, fertilize the eggs, incubate embryos, feed hatchlings and after imprinting them, release smolt into oceanic waters
to return with wild conspecifics. The PNP hatcheries cover their expenses through cost-recovery harvests. This talk will focus
on the advantages and disadvantages of expanding aquaculture of marine finfish in Alaska.

THE UTILIZATION OF HISTOPATHOLOGY IN AQUACULTURE RESEARCH AND THE
DEVELOPMENT OF A HISTOLOGIC LESION GRADING SCHEME
Kevin Snekvik*, Christopher Good, John Davidson, Carla Welsh and Steven Summerfelt
Washington Animal Disease Diagnostic Laboratory
Department of Veterinary Microbiology and Pathology
College of Veterinary Medicine
Washington State University
Bustad Hall, Rm 155N
Pullman, WA 99165-2037 USA
ksnek@vetmed.wsu.edu
While several parameters, such as overall mortality or grow rate, are typically evaluated during research experiments, histopathology
is often overlooked as an important tool in understanding the effect of various treatments, pathogen challenges or
environmental conditions on the examined species. In particular, histopathology provides a view of both the effect of specific
treatments on individual organ systems, for example epithelial cell degeneration in the gills or inflammatory cell infiltrates in
the liver, as well as providing a general assessment of the entire animal, such as muscle atrophy during anorexia, which few
other assays can provide. The response of the individual organs or the immune system can then be segregated into multiple
grades from minimal to severe to better determine the effect of the treatment. In a set of experiments utilizing rainbow trout in
recirculation systems we have included the histological evaluation of multiple organs and established a histopathological grading
scheme to evaluate individual organ systems as well as determine an overall “damage score” to each fish within a particular
treatment group. This has allowed a semi-quantitative assessment of disease between particular groups.
32

32
SELECTIVE BREEDING AND A NON-FISH MEAL DIET FOR RAINBOW TROUT
Oncorhynchus mykiss: A CLOSER LOOK AT FAMILY CX-70
Scott Snyder*, Gibson Gaylord, Ken Overturf, Frederic Barrows and Ronald Hardy
Hagerman Fish Culture Experiment Station
University of Idaho and USDA-ARS
3059-F National Fish Hatchery Road
Hagerman, ID 83332 USA
scott.snyder@vandals.uidaho.edu
Studies have shown variation in growth performance between animal strains can be accounted for by either increased feed
intake or altered metabolic efficiencies. This variability in nutrient retention efficiency allows for selective breeding programs
that can effectively create lines of rainbow trout (RBT) better able to utilize plant-derived nutrients. A RBT brood stock selection
program exists at the Hagerman Fish Culture Experiment Station (HFCES) with this goal. By obtaining founder stock
(USFWS Arlee, CSI-House Creek) green eggs and milt, and f 3 ’s of the Arlee (A) x House Creek (HC) line, we simultaneously
reared 5 families of fish; founder stock (A x A, HC x HC), a mimic of the original f 1 ’s (A x HC, HC x A) and f 3 ’s (CX-70) of
this RBT line for the purpose of conducting a growth trial to identify the changes in fish growth after 2 generations of selection
due to plant-derived nutrient selection pressure.
Utilizing a 5x2 factorial design containing 5 families of fish, 2 diets and 3 replicates per treatment, thirty 145-l flow-through
tanks were randomly assigned a treatment replicate and stocked with twenty 40-g fish each. Each tank was supplied with 15 °C
spring water with a flow rate of 12-lmin -1 . Diet 1 (Fish) (47%CP, 5400 cal/g GE, 15% CF) derived all of its protein from fish
meal and diet 2 (Plant) (44%CP, 5400 cal/g GE, 15% CF) derived all of its protein from a blend of plant products and serves
as the selection diet in the selection program at HFCES. Fish were fed to apparent satiation 2 times per day, 6 days per week
for 12 weeks.

USE OF FRESHWATER LOW VALUE FISH FOR AQUACULTURE DEVELOPMENT IN THE
CAMBODIA’S MEKONG RIVER BASIN
So Nam * , Eng Tong, Souen Norng and Kent Hortle
Inland Fisheries Research and Development Institute
Fisheries Administration
# 186, Norodom Blvd.
Phnom Penh, Cambodia
sonammekong2001@yahoo.com
Aquaculture of carnivorous and omnivorous fish species and crocodile in Cambodia is highly dependent on inland fisheries of
low value fish for sourcing key dietary nutrient inputs. Thousands of tones of low value fish are also used for producing fish
and other animal meals and for human consumption.
This study provides clear understanding of the current status of the use of freshwater low value fish in Cambodia. In order to
achieve this goal, five specific objectives are developed: (1) To assess technological impacts of use of freshwater low value fish
for aquaculture development in Cambodia; (2) To determine approximate quantities or proportions and numbers, to the extent
possible, of low value fish species used as direct feed for growing carnivorous and omnivorous fish species and crocodile; (3)
To assess socioeconomic impacts of the use of low value fish; (4) To examine actions/strategies to address the identified negative
impacts; and (5) To imply for sustainable aquaculture development and fisheries management in the Cambodia’s Mekong
basin.
In order to achieve the above objectives, primarily field surveys of the main types of aquaculture of carnivorous species (fin
fish and crocodiles) and omnivorous species (only fin fish), which are respectively mainly and partially fed with low-marketed
value fish, is carried out among local farmers in the four major river branches of the Cambodia’s Mekong River basin, with a
distance of over 500 km using developed questionnaires.
32

32
oPtimiZinG GroWtH AnD QUALitY of fArmeD coD At 67◦n
Christel Solberg
Department of Fisheries and Natural Science
Bodø University College
N-8049 Bodø, Norway
www.hibo.no
Major roads to ensure optimal production conditions for cod is either to change the feeding conditions or by changing the feed
composition.
Experiments done in North Norway showed that decreasing the number of given meals per week from five times to 2 times
a week, resulted in the same growth and the same quality of the cod with a considerable reduction in the FCR from 1.47 to
1.35.
In another experiment it was used feed from three different companies in a full scale trial, to elucidate if the different feed had
any effect on the growth or quality of the fish.
Cod juveniles with an average size of 100g produced in Vestlandet, were transported to Bodø in the end of October 2004. It was
used about 35.000 fish in each 60 m netpen. Samplings of fish were performed in January 2005, June, August, January 2006,
July and September.
The proximate composition of the feed was approximately similar in the different feed, 55-52 % protein and 15-20% fat.
However, in September 2006 significant differences in the gutted weight were found. There were not found any differences in
conditionfactor, liverindex or muscle watercontent. The fatty acid composition in the liver was different for the different feed,
indicating different sources of the oil in the feed. It was observed green spots on the liver, especially from cod fed with one of
the feeds, making the cod liver unsuitable for the commercial market. The reason for this discoloration is for the moment not
known.

GROWTH OF PACIFIC WHITE SHRIMP Litopenaeus vannamei FED DIETS CONTAINING
DIFFERENT LEVELS OF SOY PROTEIN CONCENTRATE REARED UNDER CLEAR
WATER SYSTEM
Daranee Sookying*, Patricio Paz and D. Allen Davis
Department of Fisheries and Allied Aquacultures
Auburn University
Auburn, AL 36849-5419 USA
sookyda@auburn.edu
The study on the use of soy protein products as alternative protein sources in shrimp diets has been a focus all over the world
because of their favorable protein content, suitable amino acid profile, economic opportunity, and consistent quality. Previous
research demonstrated the potential of using soy protein concentrate (SPC) at low level as replacement of solvent extracted
soy bean meal (SE-SBM) in shrimp feed. However, as high levels resulted in reduced growth further work was warranted. The
objective of this study was to confirm the growth response of Pacific white shrimp Litopenaeus vannamei fed practical diets
containing different levels of soy protein concentrate in substitution of soybean meal, when reared in the clear water condition.
Two trials were carried out at the E.W. Shell Fisheries Research Station (EWS) in Auburn, Alabama and Claude Peteet Mariculture
Center (CPMC) in Gulf Shores, Alabama.
In the first trial, six dietary treatments containing increasing percentages of SPC (0%, 5%, 10%, 20%, 40% and 40% with
additional fish soluble) were randomly assigned amongst 24 tanks (80 L) with four replications per treatment. Juvenile shrimps
(initial weight 0.12 g) were stocked at a density of 12 shrimps per tank during an 8 week feeding trial. Daily feed inputs were
calculated, based upon an expected weight gain of 0.8 g per week and an expected feed conversion ratio of 1.8:1.
In the second trial, four dietary treatments were produced under commercial conditions and the diets were randomly assigned
amongst 12 tanks (425L) with three replications per treatment. The experimental diets contained increasing level of SPC (0%,
4%, 8%, and 12%). Juvenile shrimp (0.9g initial weight) were stocked into the tanks at a density of 15 shrimp per tank during a
12 week feeding trial. Daily feed inputs were calculated, based upon an expected weight gain of 1.0 g per week and an expected
feed conversion ratio of 1.5:1.
Water exchanged was done at a rate of 100% every day with a re-circulating system containing a biological filter, and a circulation
pump throughout the trial period. Dissolved oxygen, temperature, and pH were measured twice a day, and total ammonianitrogen
was measured biweekly. At the end of the experiment, growth, survival and feed conversion ratio from both systems
will be determined and compared to examine the variation performance of Pacific white shrimp.
32

330
FEASIBILITY OF USING DIFFERENT LEVELS OF SOY PROTEIN CONCENTRATE AS
THE PROTEIN SOURCE IN FEED FOR PACIFIC WHITE SHRIMP Litopenaeus vannamei
REARED UNDER POND CONDITION
Daranee Sookying*, Jessica N. Jacquay and D. Allen Davis
Department of Fisheries and Allied Aquacultures
Auburn University
Auburn, AL 36849-5419 USA
sookyda@auburn.edu
Soybeans are one of the most widely distributed oil seeds which produce a variety of products used in the animal feed industry.
For example, solvent extracted soybean meal (SE-SBM) is one of the most efficient and widely used protein sources in the
feed industry. This is primarily due to the high quality of nutrients especially with respect to protein and amino acid profiles.
However, as compared to fish meal it has a lower nutrient content in terms of protein, essential amino acids (e.g. methionine),
essential fatty acids and minerals which can be adjusted with supplements or blending with other products. As the nutrient
density (protein and energy) of the diet is increased the use of SE-SBM is inefficient as there is insufficient room in the formulation
for all necessary components. Furthermore, even in diet with moderate levels of protein there are advantages to having a
high protein ingredient in the formulation. Soy protein concentrate (SPC) is a high protein ingredient that is produced through
a series of different extraction and precipitation process from high quality dehulled soybeans. By removing most of the oil and
water soluble non-protein constituents a variety of high protein products with varying characteristics can be produced. These
products are increasingly being used by the animal feed industry. As the availability of these product increases, the price is
likely to be reduced making them cost effective solutions. As SPC has a higher protein content than SE-SBM, there is interest
in using these meals in practical diet formulations. The objective of this project was to demonstrate the feasibility of diets
formulated to contain increasing percentages of SPC (0%, 4%, 8%, and 12%), in production diets for L. vannamei reared under
pond production conditions. The research was conducted at Claude Peteet Mariculture Center in Gulf Shores, Alabama. Four
diets are currently being tested in sixteen 0.1 Ha ponds using four replicates per diet. Post larvae were obtained from Shrimp
Improvement Systems in the Florida Keys, Florida and nursed for two weeks, juvenile shrimps (~0.01g) were stocked at 35
shrimp/m 2 and will be cultured under standardized production conditions. Feed inputs are pre-scheduled using feeding protocols
that have maximum feed inputs of 7.23 kg/ha/day. The experiment is currently ongoing and the growout stage is scheduled
to last 17 weeks. After 9 weeks of pond culture, cast net samples have an average mean weight of 10.41 grams, and all treatments
are showing similar growth rates. Final yield, weight gain, survival rate, and feed conversion ratio will be determined
and reported at the conclusion of the production cycle of the ponds after harvest. Moreover, the same diets were tested in 20
tanks (800L) with five replications per treatment, and four tanks (800L) were used with a fifth diet that served as a commercial
reference with four replicates. Juvenile shrimp (1.0g initial weight) were stocked at a density of 30 shrimp per tank. Water
was pumped from a production pond to mimic production pond conditions and water exchanges occurred daily throughout the
12 week trial. Daily feed inputs were calculated, based upon an expected weight gain of 1.3 g per week and an expected feed
conversion ratio of 1.2:1. Growth, survival, and feed conversion ratio will be determined at the end of experimental period.

DEVELOPMENT OF SMALL-SCALE AQUACULTURE; TROUT FARMING SYSTEMS IN
THE WESTERN-CAPE PROVINCE OF SOUTH AFRICA
Stander, H.B.
Aquaculture Division
Faculty of Agriculture and Forestry Science
University of Stellenbosch
Private Bag X1
Matieland 7602
South Africa
hbs@sun.ac.za
Aquaculture has the potential to contribute towards economic empowerment and social development of disadvantaged rural
communities through sustainable utilization of resources. The Division of Aquaculture at Stellenbosch University has launched
a Small-Scale Aquaculture Farming Program with the objective to improve the living standards of rural communities, provide
opportunities to participate in the regional and national economy, contribute to food security and sustainable utilization of
resources. The Program targets disadvantaged rural people with a focus on women. Various role-players including the private
sector, government departments and local authorities are participating in the Program.
The Program has been given a unique opportunity by the private sector to expand its benefits to rural communities, through
a long-term market-uptake agreement with Three Streams Smokehouse in Franschhoek. For this, the farmers must produce
a quality product according to market standards. A total of 35 additional small farmer projects can be introduced within the
framework of the market-uptake agreement.
The successful and sustainable implementation of the Program requires the provision of essential services in the form of business
development, support and training. A network of support services that include extension, training, financial, marketing,
and legal services are required to ensure effective implementation and sustainability of the small farmer projects.
This Program has proven its ability to make a meaningful contribution to the per capita income and nutritional status of these
communities, whilst it also provides a means for the development of human resources.
331

332
A PRODUCTION COST ANALYSIS OF CHANNEL CATFISH Ictalurus puntactus USING
CURRENT FEED, TRANSPORT AND ELECTRICAL COSTS
James A. Steeby and Terrill R. Hanson
National Warmwater Aquaculture Center
Box 239
Belzoni, MS 39038 USA
jsteeby@ext.msstat.edu
Research in catfish production using various protein levels has been well examined from total net harvest and harvestable fish
standpoints. Current high feed and fuel prices, including transport and electrical aeration costs were combined with a study by
Li et al. 2006 to examine how cost effective the various protein concentration feeds are when they are fed daily (D) and every
other day (EOD). The cost per net kg of fish produced for 24, 28 and 32 % protein feeds were similar when fish were fed daily
and every other day. The 32% protein feed fed daily yielded slightly higher cost per kg of harvestable size fish compared with
the other two feed protein levels. Feeding a 24% protein feed every other day and feeding a 36% protein feed daily were the
least cost effective. Partial enterprise budgets developed for these treatments indicated 24% protein feed fed daily resulted
in the highest net return though production was less than for 32% and 28% protein fed daily treatments. When feeding every
other day, the 32% protein diet had the highest net return with production being intermediate to the 36% and 28% EOD treatments.
Current high fuel prices indicate that feed delivery charges and aeration-electrical rates may be even more critical to
fish production costs in the future. This analysis reminds researchers and producers that maximum production does not always
result in maximum returns.

The acuTe ToxiciTy of copper To largemouTh bass aT differenT
alkaliniTies
Allison Stewart, Shawn Coyle, Robert Durborow and James H. Tidwell
Kentucky State University
Aquaculture Research Center
Frankfort, KY 40601 USA
ahstewar@colby.edu
Copper sulfate is used extensively in aquaculture as an algicide and as a therapeutant for protozoan parasites because of its
effectiveness and low cost, but it can be extremely toxic to fish in water having low alkalinity. The acute toxicity of copper (Cu)
of many species has been studied; however, currently there is little data available for largemouth bass.
Largemouth bass, Micropterus salmoides, juveniles (5.6±1.1 g) were exposed to copper sulfate (CuSO 4 ) in a series of static
toxicity tests to observe species sensitivity. Water used in these experiments had a series of alkalinity levels. Tests were
conducted in 3-L polycarbonate aquaria equipped with air stones to maintain dissolved oxygen levels greater than 75% saturation.
Each aquarium contained 5 fish. The experiments were conducted under static test conditions to imitate conditions in
aquaculture ponds where the compound is applied in a single dose and concentrations of copper decline after application. In
each trial, fish were exposed to a control (no CuSO 4 added) and at least six concentrations of CuSO 4 . CuSO 4 was added from
stock solutions based on a range finding studies test in a completely randomized design with three replications per treatment.
Concentrations were selected to produce 0% to 100% mortality of test animals within 48 h. The 48 h duration was chosen based
on the observed mortality pattern in preliminary studies and the rapid precipitation of Cu from solution.
Probit analysis was used to calculate estimates of 48-h LC50 values (median lethal concentration). The data demonstrated the
typical acute toxicity response of Cu, in which the toxicity to fish increases as pH, total alkalinity and total hardness decrease.
The results indicate that largemouth bass are relatively tolerant to Cu compared to other species that have been studied.
333

334
OVERCOMING OPERATIONAL CHALLENGES IN OFFSHORE FISH FARMING
R. Chris Stock*, Glenn Flanders and Steve Page
Ocean Farm Technologies
114 Higgins Rd. N.
Searsmont ME, 04973 USA
cstock@oceanfarmtech.com
Deep sea, open ocean aquaculture has become a reality only in very recent history. Modern engineering and technological advances
have helped create solutions to many challenges that were previously considered impediments to the success of offshore
farming. At the same time innovative new pen designs created for offshore fish farming have provided solutions to some more
traditional aquaculture problems.
Open ocean aquaculture can eliminate or reduce many of the obvious shortcomings of present coastal fish farms such as excess
waste deposition, disease outbreaks and common space use conflicts but it has also produced some unexpected solutions to
other equally important challenges. The success of newer more practical submersible pens can be attributed to their designs
which have taken into account the operational deficiencies of floating cages and near shore aquaculture.
Submersible pens such as the Aquapod easily incorporate rigid mesh materials like marine grade galvanized steel which were
previously difficult or impossible to apply to floating pens. These materials have proven impenetrable to predators and can
eliminate escapes which have proven to be a perennial challenge to conventional net pen fish farmers. Another disadvantage to
traditional nets is that handling them for maintenance purposes is a laborious and time consuming process. Some submersible
offshore pens can be surfaced and rotated in their moorings to allow for inspection, repair and cleaning above the waterline
which dramatically cuts costs, reduces dive times and improves safety.
Submerging these pens not only shelters them from storms and foul weather but can also allow the farmer to take advantage of
favorable environmental conditions not found at the surface. Production at depth can reduce fouling and often provides cooler
more consistent temperatures which may benefit particular species, such as cod.
Open ocean aquaculture not only provides optimal conditions for the health and growth of a crop, but it is an ideal solution to
many traditional operating problems. Present and emerging technologies will continually improve offshore fish farming and
eventually it will become a predominant means of aquaculture production.

GROWTH OF FED GOLDEN SHINERS IN AQUARIA SUPPLIED WITH SIZE-
FRACTIONATED NATURAL FOODS
Nathan Stone*, Marcella Melandri and Greg O’Neil
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601 USA
nstone@uaex.edu
Golden shiners in indoor systems exhibit relatively slow growth even when fed a complete diet. The cause is thought to be the
lack of natural foods, although this has not been confirmed. A 4-week trial was conducted in an 18, 110-L aquaria system to
determine the growth response of golden shiners juveniles to the presence of three size-fractions of natural foods. A 0.37-kW
pump submerged in a fertilized earthen pond delivered water to the aquaria via a pressurized continuous loop of pipe. Pond
water was filtered through 10- or 100-micron filters or not filtered (raw). Pressure-compensating orifices in the discharge pipe
of each filter regulated flow and each aquarium received water at a nominal rate of 3.8 L/min. Each aquarium was stocked with
20 juvenile golden shiners (average weight of 0.04 g) and fish were fed once daily with a 42% crude protein commercial flake
feed at a rate of 3% of their body weight. Zooplankton samples were collected twice weekly by filtering 30-L of inflowing water
from 2 randomly-selected aquaria per treatment.
Weight gain of fish with access to larger zooplankton (no filter) was double that of fish in the 10-micron treatment (Table 1) and
fish were in better condition. Zooplankton concentrations were highly variable within treatments and over time. Overall mean
concentrations of rotifers decreased from 398/L in the no filter treatment to 57/L and 12/L in the 100- and 10-micron treatments,
respectively, and total zooplankton concentrations (excluding rotifers) followed a similar pattern. The growth rate of fish in this
study was greater than that reported for indoor studies and comparable to that of similarly-sized fish cultured in earthen ponds,
implying that lack of natural foods is responsible for the slow growth of golden shiners in indoor studies.
33

33
NITROGEN AND PHOSPHORUS CONCENTRATIONS REQUIRED TO INITIATE
PLANKTON BLOOMS IN COMMERCIAL BAITFISH PONDS
Nathan Stone* and Alex Kachowski
Aquaculture/Fisheries Center
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601 USA
nstone@uaex.edu
Establishing desired plankton densities in ponds while avoiding unnecessary applications of nutrients remains a challenge for
commercial baitfish producers. Ground waters from wells into the alluvial aquifer near Lonoke, Arkansas, used in filling 14
commercial baitfish ponds were analyzed, as were concurrent pond water samples obtained 24-96 h after filling commenced.
Significant differences (P < 0.05, paired t-test) in the average concentrations of various parameters were found: total alkalinity
decreased from 263 to 230 mg/L, total hardness increased from 325 to 418 mg/L, sulfate increased from 91 to 216 mg/L, phosphate
decreased from 0.60 to 0.13 mg/L, and dissolved inorganic nitrogen (DIN) increased from 0.35 to 2.09 mg/L (Figure 1).
Results imply that bottom sediments contribute to DIN and that phosphorus is precipitated. Analyses of the upper 5-cm of pond
bottom sediments in shallow, middle and deep locations within each of 25 commercial baitfish ponds found that Ca, Mg, Mn, K,
Na, S and Cu concentrations were very high (upper 10%) relative to values in the literature for freshwater aquaculture ponds.
Algal bioassays using natural plankton assemblages were conducted by spiking triplicate 250-mL flasks (water volume of 50
mL) of water from 16 newly-filling ponds with 0, 1, or 2 mg/L of N (as sodium nitrate) and 0, 0.25, or 0.50 mg/L of P (as sodium
phosphate). For 11 ponds, bioassays were conducted both in the ponds (in floating racks) and in the laboratory, while the
remaining 5 were indoor assays only. Results as determined by fluorescence and chlorophyll a extraction demonstrated that in
general, addition of phosphorus (0.25 or 0.50 mg P/L) was required for maximum phytoplankton bloom development while the
N requirement was variable, ranging from 0 to 2 mg/L.

THE SAFETY OF COPPER SULFATE TO CHANNEL CATFISH EGGS
David L. Straus*, Andrew J. Mitchell, Ray R. Carter, Mathew E. McEntire, Andrew A. Radomski
and James A. Steeby
Harry K. Dupree - Stuttgart National Aquaculture Research Center
U.S. Department of Agriculture, Agricultural Research Service
Stuttgart, AR USA
Dave.Straus@ars.usda.gov
Copper sulfate (CuSO 4 ) is an economical treatment to control fungus (Saprolegnia spp.) on channel catfish eggs and is widely
used by the industry. The purpose of this study was to determine the safety of copper sulfate to channel catfish eggs when
treated at the therapeutic rate (10 mg/L), and also at 30 and 50 mg/L CuSO 4 .
Catfish were spawned on-site and egg masses were used in the study within 24 hrs. Similar intact portions (~100 g) of an egg
mass were placed into mesh baskets of individual compartments of a custom hatching trough and acclimated for 1 hr. Egg
counts (eggs/~15 g sample) were also determined for each of the egg masses to estimate number of eggs in each portion.
Eggs were treated daily until the embryos developed eyes; exchange rate of the water was 90 minutes during treatments. Water
chemistry during the study was pH 7.5, 208 mg/L total alkalinity, and 106 mg/L total hardness; water temperature was 26ºC.
When hatching was complete for all viable eggs, fry of each compartment were preserved in 70% ethanol for counting to determine
the percent hatch in each treatment.
Some fungus developed in the controls at this temperature and mean percent hatch was 40.8%. The mean percent hatch of the
10, 30, and 50 mg/L CuSO 4 was 80.1, 64.2 and 80.2%, respectively. The difference between the 10 and 30 mg/L CuSO 4 treatments
was statistically significant, while the difference was not significant between the 10 and 50 mg/L CuSO 4 treatments. The
lower hatch-rate of the 30 mg/L treatment is attributed to the random sampling within the original egg masses and the range of
hatching rates that are common in the industry. A separate experiment looked at the hatching success when eggs were treated
daily until the embryos developed eyes with 100 mg/L CuSO 4 . The water temperature was 24ºC and the exchange rate during
the treatment was 30 minutes. The individual percent hatch of each replication was 62.7, 94.9, 59.7 and 64.8%.
33

33
ACUTE TOXICITY OF PERACETIC ACID (PAA) TO Ichthyophthirius multifiliis THERONTS
David L. Straus and Thomas Meinelt
Harry K. Dupree - Stuttgart National Aquaculture Research Center
U.S. Department of Agriculture
Agricultural Research Service
Stuttgart, AR USA
Dave.Straus@ars.usda.gov
Peracetic acid (PAA) is an antimicrobial disinfectant used in agriculture, food processing and medical facilities. It has recently
been suggested as a means to control infestations of Ichthyophthirius multifiliis. The purpose of this study was to determine
the acute toxicity of two products containing 4.5% and 40% PAA to I. multifiliis theronts from two geographically separate isolates.
Theronts were exposed to concentrations of PAA in 96-well plates containing groundwater at 23°C. Acute toxicity was
observed over a 4 h period. No significant difference in the median lethal concentration (LC 50 ) estimates was evident between
the two isolates at 4 hrs with the 4.5% PAA product (0.146 versus 0.108 mg/l PAA), while there was a statistical difference
between the 4 hr LC 50 with the 40% PAA product (0.274 versus 0.158 mg/l PAA). These results suggest that PAA is toxic to I.
multifiliis theronts at low concentrations and that one of the isolates was more resistant to this compound.

PRACTICAL APPLICATION OF AN INTEGRATED LARVAL-NURSERY SYSTEM FOR THE
PRODUCTION OF CALIFORNIA HALIBUT, Paralichthys californicus
Kevin Stuart*, Michael Paquette, Keri Maull, Mark Drawbridge and Paula Sylvia
Hubbs-SeaWorld Research Institute
2595 Ingraham Street
San Diego, CA 92109 USA
kstuart@hswri.org
In order to reduce stress and mortality during transfer of larval California halibut immediately prior to settlement, we developed
a larval-nursery system that combined two of our preferred rearing vessel designs into one integrated, space-saving system
(Figure 1). The engineering aspects of this system were reported at WAS 2007. This paper reports on the first of three planned
rearing trials in this system.
Halibut eggs were collected from a 44 m 3 broodstock tank, maintained under ambient temperature and photoperiod conditions.
Egg quality parameters examined for this trial included egg viability (81.9%), hatch rate (84%) and survival to 1 st feeding
(91%). The eggs were placed into one of four 1600L black conical bottom tanks at a density of 100 eggs/L. The tanks were
maintained at 17.7 C (± 0.94 C) under ambient lighting (0 – 2200 lux) on an independent recirculating system. The flow to
each tank was set at 5 L/min.
Halibut larvae were reared in the cone tanks using traditional greenwater and rotifer and/or Artemia live feeds until 27 dph.
At 27 dph the pelagic larvae were transferred to an independent recirculating raceway system by selecting one of three 5 cm
transfer pipes that exited each cone tank at different elevations corresponding to one of three vertically stacked raceways. The
transfer process took approximately 1-2 days when transfer flows were set at 8 – 12 L/min, there were no observable effects
on fish health from transfer. Dry feed was introduced to the juveniles at 32 dph and weaning was complete by 45 dph. In the
raceways, water flow rates and depth were adjusted based on the stage of fish development from 10 – 90 L/min and 5.1-30.5 cm
deep. This created water velocities of 0.6 – 6.0 cm/sec. Water velocities were maintained at a relative rate of 1.0 to 1.5 body
lengths per second in order to facilitate self cleaning, and promote faster growth.
Halibut grew to 4.0 g by 96 dph. Survival from egg was 1.1 – 2.2% when evaluated for each raceway cohort. The percent of
malpigmented halibut was also high 20-30%. No disease outbreaks were observed. Overall the system worked very well and
as we had hoped. The most significant losses were due to aggression and cannibalism in the shallow raceways as pelagic larvae
were “harassed” by larger, settled siblings. Future rearing trials will incorporate aggressive grading protocols to minimize
these losses.
33

340
ANTIBACTERIAL ACTIVITY OF CATIONIC PEPTIDES ALONE OR IN COMBINATION
AGAINST MARINE Vibrio spp. CAUSING VIBRIOSIS IN FISH AND SHELLFISH
Bobban Subhadra*, Annabeth Fieck, Ivy Hurwitz and Ravi Durvasula
Department of Internal Medicine
Division of Infectious Diseases
University of New Mexico
Albuquerque, NM 87131 USA
BSubhadra@salud.unm.edu
Paratransgenic strategy to control infection of commercial shrimp would involve expression of antibacterial molecules in feed
organism such as Artemia. We studied the antibacterial activity of cationic peptides (apidaecin, mellitin, moricin, magainin,
cecropin and penaedin) alone or in combination against Vibrio spp., probiotic bacteria and algae for optimizing paratransgenic
disease control approach in shrimp culture. Moricin was the most potent antibacterial agent against Vibrio strains with
minimum bactericidal concentration (MBC) ranged from 0.04 µM to 0.313µM. The MBC of cecropin against Vibrio spp.
ranged from 0.02-17.5 µM. The MBC of mellitin against Vibrio spp. ranged from 2.5-10.0 µM. The MBC of both penaeidin
and apidaecin against all the Vibrio spp. and non-Vibrio spp. were more than 20µM. Brevibacterium linens, Synechococcus
bacillarus, and Dunaliella salina had high MBC (>20µM) compared to all other bacteria tested. Moricin was the only peptide
which had efficient killing activity against B. linens, S. bacillarus, and D. salina. Combination of moricin and mellitin (MO +
ME) had the highest synergistic antibacterial activity against Vibrio spp. (Table 1). Antibacterial experimental selection studies
showed that Vibrio harveyi, Vibrio penaeicida, and Vibrio campbelli evolved resistance towards mellitin. The study showed
that moricin, mellitin and cecropin have potent antimicrobial activity against Vibrio spp. and these peptides can be expressed in
B. subtilis and algal species like S. bacillarus and D. salina as a paratransgenic approach to control vibriosis in intensive shrimp
aquaculture.
TABLE 1. Effects of combination of antibacterial peptides against
a, b
bacteria using a disc diffusion method
Combination
Number out of 12 Vibrio strains tested
Synergy Weak synergy No synergy
CE + ME 3 9 0
CE + MO 4 8 0
CE + MG 6 6 0
MO + ME 7 5 0
ME + MG 5 7 0
MO + MG 6 6 0
a Synergy = well characterized change of 2 mm in the inhibition zone;
Weak synergy = change of 2 mm in the inhibition zone;
No synergy = no change observed in the inhibition zone.
b Abbreviated as follows: CE + ME= cecropin + mellitin;
CE + MO = cecropin + moricin; CE+MG = cecropin + magainin;
ME+ MG = mellitin + magainin ; ME+MO = mellitin + moricin,
MO+MG = moricin + magainin

DEVELOPMENT OF PARATRANSGENIC Artemia: A POTENTIAL STRATEGY FOR
CONTROL OF INFECTIOUS DISEASES OF COMMERCIAL MARICULTURE
Bobban Subhadra*, Ivy Hurwitz, Annabeth Fieck, D.V. Subba Rao, G. Subba Rao and Ravi Durvasula
Department of Internal Medicine
Division of Infectious Diseases
University of New Mexico
Albuquerque, NM 87131 USA
BSubhadra@salud.unm.edu
Commercial rearing of shrimp constitutes a major part of global aquaculture. Outbreaks of infectious diseases such as White
Spot Syndrome Virus (WSSV) and vibriosis have repeatedly devastated the mariculture industry in many countries. Given the
tremendous global impact of WSSV and vibriosis on shrimp farming and the constraints of high-intensity cultivation, new
strategies to combat these diseases are essential. Our laboratory has developed a novel approach to control infectious disease
transmission, termed paratransgenesis. In this strategy, commensal or symbiotic bacteria found at mucosal sites of pathogen
transmission are isolated and genetically altered to elaborate molecules that either kill infectious agents or disrupt transmission.
We initially developed paratransgenic approaches to control transmission of certain arthropod-borne diseases. Here we
report application of the paratransgenic approach to infectious diseases of commercial shrimp mariculture.We studied the
accumulation and retention of recombinant proteins in Artemia gut for optimizing paratransgenic disease control in shrimp
culture. Transgenic Escherichia coli expressing fluorescent marker proteins and the transgenic cyanobacterium, Synechococcus
bacillarus expressing a functional murine single chain antibody, DB3, were fed to Artemia. Stable expression and retention of
several marker molecules (e.g. GFP DS red and DB3) up to 20 h post feeding were evident within the gut of Artemia (Figure
1). Engineered strains of S. bacillarus expressing DB3 accumulated within the gut of Artemia, with detectable antibody activity
for 8 h post-feeding via ELISA, coincident with the time period of highest density of transgenic S. bacillarus in the Artemia
gut. Artemia fed transgenic bacteria or algae accumulated recombinant proteins for up to 20 h which retained biological
activity. Co-delivery of multiple recombinant proteins simultaneously in the gut of Artemia was also demonstrated. The
simultaneous delivery, expression and retention of multiple recombinant molecules (e.g. antibodies and antibacterials) directed
against infectious agents (e.g. Vibrio spp. and white spot virus) in Artemia and subsequent feeding of this Artemia to shrimp are
key elements in the paratransgenic disease control approach.
Figure 1 Accumulation of
recombinant proteins in 2 nd instar
naupllii of Artemia fed with transgenic
E. coli. a) Artemia fed with wild type
E. coli after 6 h of feeding b) Artemia
fed with GFP expressing bacteria after
30 min of feeding c) 4 h of feeding d)
10 h of feeding e) Artemia fed with DS
red expressing bacteria after 6 h of
feeding f) Artemia fed simultaneously
with both GFP expressing bacteria and
DS red expressing E. coli after 6 h of
feeding.
341

342
EFFECTS OF SALINITY ON GROWTH AND PLASMA OSMOLALITY OF JUVENILE
ALLIGATOR GAR Atractosteus spatula
Mark D. Suchy, Timothy A. Clay, Wendell J. Lorio, Allyse M. Ferrara and Quenton C. Fontenot
Bayousphere Research Laboratory
Nicholls State University
Thibodaux, LA 70310 USA
Suchy6@hotmail.com
Juvenile alligator gar Atractosteus spatula were exposed to various salinities to determine the effects on growth and plasma
osmolality. After acclimated to treatment salinity levels (0, 4, 8, or 12 ppt), fish were stocked at 5 fish/L in 60 L recirculating
systems (28.1±2.1 °C) and reared for 31 d in a greenhouse. Fish were fed floating feed (45% protein/16% lipid) daily either
10% body weight {19-33 days after hatch (DAH)} or 5% body weight (34-51 DAH). Growth was greater (α=0.05) at 4 and 8
than 0 and 12 ppt (Fig. 1)
To determine the ability of juvenile alligator
gar to maintain plasma osmolality (mOsm), we
exposed fish to salinities ranging from 0 to
37 ppt with or without an acclimation period.
Plasma osmolality was measured for three
un-acclimated fish (maintained at 0 ppt)
after 24 hr exposure to either 0, 4, 8, 12, 16,
20, 24, 28, or 32 ppt. Compared to the 0
ppt treatment (305±9.8), plasma osmolality
levels were elevated for the 16 ppt (337±17.0)
and higher treatments with one mortality at
28 ppt and two at 32 ppt. No mortalities
occurred during the acclimation trial where
salinity was increased from 0 to 37 by 1
ppt per day. Plasma osmolality levels were
elevated by the end of the study (37 ppt:
372.3). Un-acclimated fish had higher plasma
osmolality levels at 8 ppt (aclimated; 304±1.3:
un-acclimated; 313±3.1) and greater salinities
(Fig. 2). Alligator gar are euryhaline and
grow faster at 4 and 8 than 0 and 12 ppt.

EFFECTS OF LOW-PHOSPHORUS DIETS ON BODY FAT CONTENT, FATTY ACID
COMPOSITION, AND LIPOLYTIC GENE EXPRESSION IN RAINBOW TROUT
Shozo H. Sugiura
School of Environmental Sciences
University of Shiga Prefecture
Hikone, Japan
sugiura@ses.usp.ac.jp
Fish feeds low in phosphorus (P) have been used widely in environmentally
friendly aquaculture. Such dietary regimens sometimes, though subconsciously,
induce marginal P deficiency in fish. P-deficient fish are known to
increase body fat deposition. However, the mechanism has not been well
researched. The present study examined the effect and mechanism of fat
deposition with low-P feeds. The ultimate aim of study was to increase
commercial and nutritional values of aquaculture fishes.
One hundred rainbow trout (mean BW 21.1g) were randomly stocked
into four 50L aquaria. Four test diets, differing in P content (low-P: LP
or high-P: HP) and fat source (fish oil or lard), were made as dry pellets
(Table 1), and hand-fed to fish twice daily to satiation for 2 months. At
the end of experiment, samples were taken (5 fish /tank) after 24h of
fasting for analyses. Fillet fatty acids were analyzed by GC, and hepatic
gene expressions were determined by realtime RT-PCR.
Bone P% of fish were 3.8, 8.9, 5.1, 9.0, respectively for LP/fish oil,
HP/fish oil, LP/lard, HP/lard diets. Fish fed LP diets reduced not only
bone P% (p

344
PROCESSES TO IMPROVE ENERGY EFFICIENCY DURING PUMPING AND AERATION
OF RECIRCULATING WATER IN CIRCULAR TANK SYSTEMS
Steven T. Summerfelt, Timothy Pfeiffer and Dane Schiro
Conservation Fund Freshwater Institute
1098 Turner Road
Shepherdstown, West Virginia 25443 USA
s.summerfelt@freshwaterinstitute.org
Conventional gas transfer technologies for aquaculture systems occupy a large amount of space, require a considerable capital
investment, and can contribute to high electricity demand. In addition, diffused aeration in a circular culture tank can interfere
with the hydrodynamics of water rotation and the speed and efficiency of solids fractionation to the tank’s bottom-center
drain. To improve the energy efficiency of pumping and aerating water in circular tank-based recirculating systems while
maintaining culture tank hydrodynamics, two processes were developed that provide high water flow and low lift method of
gas exchange. One process incorporates a sidewall box airlift pump; the other process incorporates a propeller aerator mounted
at the top of the riser chamber in the sidewall box.
Two different sidewall box airlift pumps were attached to the 1.2 m tall fiberglass wall panel of a 3.7 m diameter circular tank.
Both airlifts were used to create a simple partial reuse system. The better airlift (2 nd unit tested) supported food-size rainbow
trout (Oncorhynchus mykiss) at a density of 80 kg/m 3 and a feed rate of 0.8% body weight per day without using supplementary
oxygen beyond that supplied by the make-up water. In the first trial, air was supplied with a 0.25 kW (1/3-HP) blower through
three coarse-bubble cap diffusers at a submergence depth of 0.86 m, i.e., at the base of the airlift chamber. The second airlift
was supplied by a 0.38 kW (1/2-HP) blower and 2.2 m (7 feet) of Aero-Tube aeration tubing at a depth of 0.94 m below the
water surface. Water velocity measurements taken as the water flowed horizontally from the down-welling to the up-welling
side at the base of the airlift indicate that the first airlift pump generated 1.7 m 3 /min of water flow at an air:water (vol:vol) of
approximately 0.06; the second airlift moved 1.9 m 3 /min of water flow at an air:water of < 0.02. However, the first airlift pump
increased the water level at the top of the up-welling chamber by approximately 5 cm, but the second airlift generated 3.8 cm
lift. Across the 1 st airlift, 0.45 mg/L dissolved oxygen (DO) was added while 1.6 mg/L dissolved carbon dioxide (CO 2 ) was
removed. The 2 nd airlift added 0.99 mg/L DO and removed 1.79 mg/L CO 2 . These aeration tests were conducted at 13ºC and
an inlet dissolved oxygen concentration of approximately 7.0 mg/L. The 1 st airlift created a higher flow per kW, i.e., 3.9 m 3 /min
per kW compared to 2.9 m 3 /min per per kW of the second airlift. However, the 2 nd airlift provided more efficient oxygen transfer,
i.e., 0.18 kg O 2 per kWh compared to 0.11 kg O 2 per kWh. Thus, the primary purpose of this design (gas exchange) is more
efficiently accomplished by the 2 nd airlift even when water movement is lower. Total suspended solids (TSS) fragmentation did
not appear to be an issue as the TSS concentration in the tank averaged below 1 mg/L for both airlifts. The sidewall box airlift
technology provided relatively energy efficient pumping, gas exchange, and water rotation in circular tanks.
Results (pumping rate, oxygen and carbon dioxide transfer efficiency, and energy requirements) from the second sidewall box
aeration pump process, which incorporates a propeller aerator at the top of the riser chamber in the sidewall box, will also be
reported.

TREATMENT OF RECIRCULATING WATER USING UV IRRADIATION ONLY OR A
COMBINATION OF OZONATION AND UV IRRADIATION
Steven T. Summerfelt, Mark Sharrer, Scott Tsukuda and Michael Gearheart
The Conservation Fund’s Freshwater Institute
1098 Turner Road
Shepherdstown, WV 25443 USA
s.summerfelt@freshwaterinstitute.org
Obligate and opportunistic fish pathogens can accumulate in recirculating aquaculture systems (RAS) that do not use a continuous
water disinfection process, especially during a disease outbreak when the micro-organism is propagating and shedding
from its host. Ozonation and ultraviolet (UV) irradiation processes have been used separately or in combination to treat water
in RAS before it returns to the fish culture tanks. The objective of the present study was to compare bacteria inactivation and
overall water quality produced in a RAS that includes either full-flow UV irradiation, a combination of ozonation and UV irradiation,
or no disinfection process (i.e., control). These respective test conditions were maintained for 12, 5, and 7 weeks in
a full-scale recirculating system located at the Freshwater Institute. System replication was not practical. During the O3 + UV
trial, a proportional integral feed back control loop was used to automatically adjust O 3 generator output to produce an oxidation
reduction potential (ORP) value of 447 ± 5 mV in the full recirculating flow exiting the 1.5 min contact tank, i.e., immediately
before the water entered the operational UV irradiation unit. The UV channel unit irradiated 100% of the 4,640 L/min
recirculating water with a dose of approximately 100 mJ/cm 2 just before the flow returned to the fish culture tank. Results
are tabulated below. The combination of O3 + UV produced complete inactivation of total heterotrophic bacteria plate counts
(i.e., 0 ± 0 cfu/ml), and nearly complete inactivation of total coliform bacteria counts (i.e., 1 col/100 ml) before the water was
returned to the culture tank. Achieving this level of treatment required adding a mean ozone dose of approximately 29 ± 3 g
O 3 per kg feed and dosing approximately 0.39 mg/L of ozone gas into the recirculating flow. Applying UV irradiation alone
reduced total heterotrophic bacteria plate counts to 5 ± 2 cfu/ml and total coliform plate counts to 112 ± 85 col/100 ml, which
was not as effective as O3 + UV. The best water quality was produced in the O3 + UV treatment. The TSS of water exiting the
UV channel averaged 8.2 ± 1.4, 4.4 ± 0.9, and 3.7 ± 0.8 mg/L, respectively, for the control, UV only, and O3 + UV treatments.
For the same three treatments, water color (and %UVT) exiting the UV channel averaged 12 ± 1 (89 ± 1%), 16 ± 2 (85 ± 1%),
and 3 ± 1 Pt-Co units (94 ± 1%), respectively.
34

34
REPLACEMENT OF FISHMEAL BY A BLENDED ANIMAL PROTEIN CONCENTRATE IN
Litopenaeus vannamei DIETS
A. Victor Suresh*, Alberto J.P. Nunes, George W. Chamberlain and Steven Gately
Integrated Aquaculture International
3303 West Twelfth Street
Hastings, NE 68902-0609 USA
victors@integratedaquaculture.com
The study addresses Goal 5 in the PPA Strategic Research Plan, specifically, approaches to practical formulation with alternative
ingredients to fishmeal. The study evaluated the performance of L. vannamei fed diets that had fishmeal replaced by a commercial
animal protein concentrate (Propak Plus, H.J. Baker & Bro., Inc. Westport, Connecticut, USA). Propak Plus (PPP) is
a protein concentrate composed of a blend of prime-grade low ash poultry meal, flash-dried blood meal, menhaden fish meal,
DL-methionine, and lysine sulfate. Unlike typical fishmeal replacement studies that analyze the effect of graded replacement of
crude protein in fishmeal by the replacement protein, this study sought to balance many more nutrients that are affected when
fishmeal is replaced.
A total of four diets were designed. Fishmeal replacement by PPP was achieved by specifying maximum levels of PPP in each
formula. In the control formula called PPP0, PPP was not offered, so this formula used prime-grade anchovy fishmeal as the
only animal protein. In the formula called PPP5, PPP was offered at a maximum level of 5%. In PPP12, PPP was allowed to
be included at its maximum subject to the restrictions of nutrient levels specified in the formula. In this formula, PPP inclusion
maxed out at 11.84%. Reduction in the specifications for essential fatty acids increased PPP addition to a maximum of
20.2%.
Shrimp with initial average weight of 3.55 ± 0.12 g were stocked in 1000-L outdoor tanks at a density of 60 shrimp/m 2 and were
fed twice daily for 72 days. There was no significant difference among shrimp fed the different diets in terms of survival (92.6
± 0.05%), yield (599 ± 80 g/m 2 ) and FCR (2.08 ± 0.25; P > 0.05). Shrimp fed PPP0, PPP5, PPP12 and PPP20 diets reached
average final weight of 15.31±0.25, 14.93±0.36, 14.68±0.16, 13.64±0.54 g, respectively. There were significant differences
(P

A LOW COST SMALL SCALE PRODUCTION SYSTEM FOR PENAEID SHRIMP IN LOW
SALINITY ARTIFICIAL SEAWATER
James P. Szyper
University of Hawaii Sea Grant College Program
875 Komohana St.
Hilo, HI 96720 USA
jszyper@hawaii.edu
A low cost, small scale system for holding and growout production of marine shrimp has been developed and tested with white
shrimp Litopenaeus vannamei in a pilot setting on a collaborating fish farm in East Hawaii Island. Low cost appropriate technology
concepts include a biofilter built of readily available materials, directed water flow produced by simple air lifts, and
low salinity artificial seawater of a low cost formula made from readily available commercial materials. Use of low salinity
artificial seawater makes the system independent of access to pumped natural seawater and minimizes salt discharged with
the minimal water exchange required. The original concept included powering the small air blower with a solar photovoltaic
system, which remains possible, but the trial was run with utility power.
The trial indicated numerous potential improvements to management practices and design revisions; on-farm trials of improved
systems are to follow. The system has potential value as a readily-isolated holding facility and growout system for shrimp or
other products.
34

34
USE OF MICROENCAPSULATED DIET FOR PACU Piaractus mesopotamicus LARVAE
Olívia C. C. Menossi, Manuel Yúfera, Rodrigo Takata, Maria I. Sánchez-Amaya , Thiago M. de Freitas,
Natalia de J. Leitão and Maria Célia Portella*
Sao Paulo State University - Aquaculture Center
14.884-900, Jaboticabal, SP, Brazil
portella@caunesp.unesp.br
A critical problem for the rearing of altricial larvae is the use of formulated diet and also the knowledge of the best time for
its introduction. The pacu Piaractus mesopotamicus is an important fish in South America and research has been carried to
improve the feeding management of the species, particularly during the larval phase. Amongst the different types of formulated
diet, the microcapsulated diet has been recognized mainly due to its capacity of nutrients retention. In this study we have evaluated
the use of three different formulated diets as first exogenous feed and a protocol to wean pacu larvae. Four-day old pacu
larvae (12 larvae/L) were fed for 22 days, five times a day. The treatments were: ART: Artemia nauplii in increased amount
during the experiment (positive control); S: No food was provided and larvae starved (negative control); DI: exclusively commercial
diet NRD1.2/2.0 (Inve, USA) throughout the experiment; DP: exclusively Poli-Peixe diet 450F (PoliNutri, Brazil);
DM: experimental microencapsulated diet throughout the experiment; and A6DI, A6DP and A6DM: Artemia nauplii from the
4 th to 9 th days after hatching (DAH), co-feeding (Artemia nauplii + the respective formulated diet) from the 10 th to 15 th DAH,
and only the formulated diet after this period.
Amongst the treatments that have received the formulated diets as exclusive food, only few larvae from the treatment DP were
alive at the end of experiment even thought in the lowest survival rate (P

DETERMINING MORPHOLOGICAL AND BIOCHEMICAL PARAMETERS ASSOCIATED
WITH EARLY OVARIAN FOLLICULAR ATRESIA IN WHITE STURGEON FEMALES
Acipenser transmontanus
Mariah J. Talbott*, Joel P. Van Eenennaam, Javier Linares-Casenave, Serge I. Doroshov, Christopher S. Guy
and Molly A.H. Webb
Montana Cooperative Fishery Research Unit
Montana State University
Bozeman, MT 59715 USA
mtalbott@montana.edu
In order to improve quality and yield of caviar in farmed white sturgeon, it is essential to correctly assess stage of ovarian
maturity and avoid harvesting females with atretic ovarian follicles. To detect atresia by changes in blood plasma parameters,
individual females (N=11) in the late phase of oogenesis were repeatedly bled and their ovaries biopsied before and after
onset of ovarian atresia. Follicular atresia was induced by transferring females at Sterling Caviar, LLC, California from cold
(10–13°C) to warm water (20°C). Follicle diameter increased and oocyte polarization indices decreased over time. Plasma
testosterone and estradiol concentrations in fish with normal follicles were significantly higher (p>0.05), compared to fish
exhibiting early histological signs of follicular atresia, such as structural changes in the egg coat. Total plasma protein and
calcium concentrations did not differ (p>0.05) between fish with normal and regressing ovaries. Results of statistical models
predicting the probability of follicular atresia based on plasma sex steroids will be presented. Our study may benefit sturgeon
farms and sturgeon hatcheries, by improving techniques for detection of ovarian atresia in the late phase of oogenesis, sensitive
to environmental and management stresses in sturgeon.
34

3 0
MOLECULAR DIVERSITY AND EVOLUTION OF TAURA SYNDROME VIRUS (TSV)
Kathy Tang*, Joel Wertheim, Solangel Navarro and Donald Lightner
Department of Veterinary Science and Microbiology
University of Arizona
Tucson, AZ 85721 USA
fengjyu@u.arizona.edu
Taura syndrome virus (TSV) is a pathogen of Litopenaeus vannamei, one of the most important species in shrimp aquaculture.
The virus was discovered in Ecuador in 1992. It had spread to the US by 1994, to the SE Asia by 1998, to Belize by 2001, and
to Venezuela by 2003. In general, RNA viruses exhibit high mutation rates; and, subsequently, high variation in protein and
nucleotide sequences occurs among TSV isolates. With 83 isolates, from 16 countries, collected from 1992 to 2008, we found
up to 10 % genetic variation in nucleotide sequence, 8.0% for amino acid sequence. The sequence information is valuable in
that it can provide insight into the spread and evolution of TSV.
To determine the evolutionary history of this virus, we performed a phylogenetic analysis using BEAST (Bayesian Evolutionary
Analysis Sampling Trees) with these 83 TSV isolates. This revealed four distinct genetic lineages: Americas (including
South/Central America, Mexico and US), Belize, SE Asia, and Venezuela; stemming from the Ecuador isolate, the ancestral
genotype. In addition, the date that new isolates were discovered corresponded with the mutation-estimate. In the US, 6 cases
of TSV were found, three from Hawaii and three from Texas. The first two occurrences (1994, 1995) were found in Hawaii, in
which cases the virus was likely introduced from South/Central America. The third outbreak of TSV was in Texas in 1996, and
this isolate is closely related to the isolates from Mexico. The fourth case, also in Texas, occurred in 1998; but, in this case, the
isolate was most closely related to isolates from Ecuador. The fifth TSV outbreak occurred in Texas in 2004, and this isolate
clustered with those from SE Asia, strongly suggesting this virus was introduced either through the value-added processing
of imported shrimp commodities from SE Asia, or through the importation of post-larvae and/or brood-stock from the same
sources that supply producers in SE Asia. The sixth outbreak occurred in Hawaii in 2007, and the isolate was most closely
related to TSV isolates from Mexico.
Although the virus genetic diversity was low, the mean mutation rate is 2.5 x 10 -3 substitutions/site/year, approximately 3 times
higher than those of most animal viruses (an average of 7.0 x 10 -4 substitutions/site/year, from 50 animal RNA viruses). The
rate of substitution increased exponentially over the 16-year span of this study. This suggests that TSV evolves more rapidly
than most RNA animal viruses.

DIETARY MAGNESIUM AND GROWTH OF PACIFIC WHITE SHRIMP Litopenaeus vannamei
IN LOW-SALINITY WATER
Kathy FJ. Tang * , Brendan G. Ambrose, Megan K. Kudera, Mario Hernandez-Acosta and Stephen G. Nelson
Department of Veterinary Science and Microbiology
University of Arizona
Tucson, AZ 85721 USA
fengjyu@u.arizona.edu
Two greenhouse trials were conducted with juvenile Pacific white shrimp Litopenaeus vannamei acclimated to low salinity (5
ppt) to compare their growth in response to magnesium (Mg) supplements to a commercial shrimp diet. Three diets, differing in
Mg content were used. A commercial shrimp diet was supplemented with MgCl 2 so that the final Mg contents of the diets were
either 0.26, 0.37, or 0.46%. One trial involved marking individual shrimp, which differed in initial size; and the second trial involved
shrimp that were of uniform size at the beginning. For the first trial, the specific growth rates were compared, and in the
second trial, final weights were used in the analysis. In both trials there was a statistically significant effect of diet Mg content
on growth, with depressed growth in the groups fed diets with higher levels of Mg. At the end of the second trial, compositional
analyses of individual shrimp were made for Ca, K, Mg, Na, and P. None of these differed significantly among diet treatments;
but for the pooled data, there was a significant relation between Mg and Ca in the samples. Under the conditions of this trial, a
recirculating raceway with low salinity water, supplementing the commercial diet with Mg did not improve shrimp growth.
Fig 1. Box plots showing the effect of Mg in prepared
diet on the growth of juvenile Pacific white shrimp,
Litopenaeus vannamei.
3 1

3 2
FEASIBILITY OF FARMING TILAPIA IN THE COOLER ENVIRONMENTS AND THE
EFFECTS OF NUTRACEUTICALS IN IMPROVING THEIR HEALTH
Jennifer Taylor, Ashley Parish and Ahmed Mustafa
Department of Biology
Indiana University Purdue University Fort Wayne
Fort Wayne, IN 46805 USA
Experiments were conducted to investigate the feasibility of farming tilapia in the cooler environments and the effects of nutraceuticals
in improving their overall health. Fish were divided and held in two different environments: warm water (26 ± 2 o C;
80 ± 5 o F) and cool water (15 ± 2 o C; 60 ± 5 o F). Within each environment, half of the fish were provided with nutraceutical
supplemented feed (either vitamin C, zinc, or combined vitamin C and zinc) and the other half of the fish were provided with
regular, commercial feed. Fish held in warm water (control) had better growth, physiological state and immune response than
fish held in cool water, although fish in cool water did not show any differences in condition factor. Fish receiving vitamin C
supplemented feed (700 mg/kg) or zinc supplemented feed (30 mg/kg) or a combination of vitamin C and zinc supplemented
feed (700 mg/kg vitamin C and 30 mg/kg zinc) were found to have improved immune response (respiratory burst and wound
healing; p

EVALUATION OF WATER QUALITY FOR LIVE TRANSPORT OF BAIT AND
ORNAMENTAL FISH FROM ARKANSAS
Hugh K. Thomforde
University of Arkansas at Pine Bluff
Fish Health Diagnostic Services
Lonoke Agricultural Center
P.O. Box 357
Lonoke, Arkansas 72086 USA
hthomforde@uaex.edu
Methods used for live transport on long-haul road vehicles are described. Golden shiners (Notemigonus crysoleucas), goldfish
(Carassius auratus) and fathead minnows (Pimephales promelas) are transported live, from farms in Arkansas to regional
distributors located throughout north America. They are marketed as sportfish bait through retail bait shops. Goldfish are also
marketed as ornamental fish in pet stores. Most transport is by road, on specialized 18-wheel vehicles fitted with modular insulated
tanks systems supported by liquid oxygen.
To prepare the fish for transport they must be
harvested several days in advance of shipment. They
are held, without feeding, in well-water with 0.2
percent sodium chloride to purge them of digestive
and urinary metabolites. Purged fish ship successfully
because they release little ammonia to transport
water. However, all water quality parameters
degrade during transport. Truck drivers use a thermometer,
dissolved oxygen meter, and pH pen to manage
conditions. Major stressors experienced by fish
during transport are low dissolved oxygen, steady
accumulation of ammonia, and carbon dioxide from
respiration, and lowered pH consequent to carbon
dioxide increase. Despite purging, healthy shiners
monitored during long-haul traport experienced the most
significant water-quality degradation during the first
hours of transport, usually because of poorly adjusted
oxygen metering, uncontrolled pH fluctuations,
and stressful increases in temperature, ammonia, and
carbon dioxide. Electronic recording devices were
used to monitor temperature, oxygen and carbon
dioxide. Ammonia was measured from samples
collected periodically during transport. Appropriate
methods of handling, harvesting and loading reduce
physical injury, reduce stress and shock, and result in
improved water quality during transport.
3 3

3 4
EVALUATION OF GROWTH PERFORMANCE ON POND-RAISED SUNSHINE BASS
WHEN FED PRACTICAL DIETS WITH REDUCED PROTEIN LEVELS AND CONTAINING
A COMBINATION OF POULTRY-BY-PRODUCT MEAL AND SOYBEAN MEAL AS TOTAL
REPLACEMENT OF MENHADEN FISH MEAL
Kenneth R. Thompson*, Linda S. Metts and Carl D. Webster
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
ken.thompson@kysu.edu
Since protein is generally the most expensive component in aquaculture diets, and fish meal (FM) is one of the most expensive
macro-ingredients (used in high percentages), a 459-day feeding trial was conducted with juvenile (mean individual weight
35 g) sunshine bass (Morone chrysops X M. saxatilis), to be grown to market-size phase-III fish, to evaluate growth, survival,
and feed efficiency when fed practical diets containing soybean meal (SBM) and poultry by-product meal (PBM) as complete
replacements for menhaden FM and simultaneously reducing dietary protein (32%, 36%, and 40%) compared with fish fed a
commercial high-quality finfish diet containing 30% FM, 40% SBM, and 40% crude protein (CP). The feeding trial was conducted
in 12, 0.04-ha earthen ponds and fish were stocked at a rate of 300 per pond.
When analyzed from the beginning of the study until harvest, there were no significant (P > 0.05) differences in mean final
weight, percentage weight gain, specific growth rate (SGR), percent survival, diet fed, and feed conversion ratio (FCR) among
treatments, and averaged 771 g, 2103%, 0.68%/day, 94.3%, 1589.5 g/fish, and 2.16, respectively (Table 1).
Results from the present study indicate that pond-grown sunshine bass can be fed a diet with 32% CP and formulated without
menhaden fish meal (FM) and no adverse effects on growth, survival percentage, and feed conversion ratio. This is the first
study to report that complete substitution of menhaden FM is possible when sunshine bass are grown in ponds to market-size
for a period 459 days when alternative protein sources, i.e., SBM and PBM are provided in the diet, and simultaneously reduce
dietary crude protein levels. Results from this study may help reduce diet costs for sunshine bass producers and feed mills,
thereby increasing profitability.

THE EFFECTS OF SOYBEAN OIL, FLAXSEED OIL AND A DAIRY-YEAST PREBIOTIC ON
THE GROWTH AND HEALTH OF CHANNEL CATFISH Ictalurus punctatus FINGERLINGS
AT LOW TEMPERATURE
Miles Thompson* and Rebecca Lochmann
Aquaculture and Fisheries Research Center
University of Arkansas at Pine Bluff
Pine Bluff, AR 71601 USA
mthompson@uaex.edu
Winter-feeding is a historically controversial and poorly understood aspect of commercial catfish production. Catfish fingerlings
fed a standard diet during mild winter periods, when water temperatures exceed 12.7 °C, tend to maintain body weight or
may even grow slightly. Since the catfish immune response is closely linked to both temperature and dietary lipid, varying the
lipid composition of the diet (via source or level) during winter-feeding schedules may have beneficial results on both growth
and overall health. The addition of functional feed additives during winter-feeding regimes may further enhance disease resistance
or growth of catfish fingerlings.
A 12-week trial was conducted using channel catfish
fingerlings (initial mean weight of 61.4 g/fish ± 1.5 g SD)
maintained at a temperature of 16°C in three separate
recirculating systems, each consisting of four 1140-L
tanks. Each diet was made using a commercially available
32%- protein pelleted basal diet, ground to 1mm and repelleted
into a sinking feed with the addition of 2% soybean
oil (control - SBO), 2% flaxseed oil (FLAX), or 2% soybean
oil and 2% dairy-yeast prebiotic (PREB). Fish were
fed once daily to apparent satiation. Feed consumption,
water temperatures and mortalities were recorded daily.
Fifty-fish sub-samples were weighed every three weeks to
monitor growth and adjust the feed rate. Nine-week results
are shown in Figure 1.
At 12 weeks, fish will be maintained on the diets for an
additional two weeks. Individual fish from each tank will
be used for health assays (alternate complement activity,
lysozyme activity, hematocrit and hemoglobin), hepatosomatic
index, and proximate and fatty acid analysis. Final
results will be presented at the meeting.
3

3
A REVIEW OF LIPID NUTRITION RESEARCH FOR THE LARGEMOUTH BASS
(Micropterus salmoides)
James H. Tidwell*, Shawn D. Coyle and Leigh Anne Bright
Aquaculture Research Center
Kentucky State University
103 Athletic Road
Frankfort, KY 40601 USA
james.tidwell@kysu.edu
The largemouth bass (Micropterus salmoides) is the largest member of the North American family of Centrarchidae. Research
on this species has primarily focused on hatchery production to sizes of ≤ 5 cm. In recent years interest has increased in the
development of methods for growing largemouth bass to larger sizes, especially for live sales into ethnic Asian Markets. However,
when compared to its economic importance, and high percentage of production cost accounted for by feed costs, relatively
few studies have addressed largemouth bass nutrition with even fewer being devoted to lipid nutrition.
An early study by Tidwell et al. (1996) suggested that for a warmwater, freshwater species, largemouth bass might require
relatively high levels of polyunsaturated fatty acids (PUFA). Levels of DHA (22:6n-3) in the lipids of the bass egg and muscle
tissues were >290% higher than levels measured in the diets, possibly indicating relative importance. In a subsequent study,
Coyle et al. 2000 replaced menhaden fish oil in the diets with squid oil to increase concentrations of PUFA and HUFA, and
especially DHA (22:6n-3). Although there was no increase in growth or feed conversion efficiency, fish fed the diet high in
DHA had significantly lower (P ≤ 0.05) whole body lipid levels and significantly higher (P ≤ 0.05) protein levels than fish fed
the other three diets.
Two more recent studies evaluated the use of lipids from different sources to replace fish oil in largemouth bass diets. Subhadra
et. al. (2006) evaluated replacement of menhaden oil with either canola oil, chicken oil, or a menhaden + chicken blend. In fish
fed canola or chicken oils body HUFA concentrations were reduced but performance was not different from fish fed the fish
oil diets. However, growth was low relative to fish fed commercial diets. Another study (Tidwell et al. 2007) also compared
alternative lipid sources including corn oil, sunflower oil (higher oleic), linseed oil and specialty oils including algal oil (high
22:6n-3) and fungal oil (high 20:4n-6). At the end of the study period, there were no significant differences (P > 0.05) between
treatment groups in terms of survival (98%), weight gain (595%), specific growth rate (2.2% per day), feed conversion ratio
(1.5), percent protein deposited (28%), or hepatosomatic index (2.3). Whole-body proximate composition was not significantly
affected (P > 0.05) by sources of added lipid, but whole- fatty acid composition showed large differences and primarily
reflected the fatty acid compositions of added oils. Largemouth bass may be able to use diets containing vegetable- and animalsource
lipids, which are less expensive than the previously recommended fish oil.

REPLACEMENT OF FISH MEAL WITH SOYBEAN MEAL IN DIETS FOR SNAKEHEAD
Channa striata
Tran Thi Thanh Hien, Chong M. Lee and David A. Bengtson
College of Aquaculture and Fisheries,
Can Tho University
Can Tho, Vietnam
ttthien@ctu.edu.vn
Culture of snakehead, Channa striata, is a rapidly growing industry in the Mekong River in Vietnam and Cambodia, but relies
largely on trash fish for feed. In order to make the industry more sustainable, we seek to develop snakehead diets based on fish
meal and plant proteins. In this experiment, we tested a control diet of fish meal against diets in which soybean meal replaced
fish meal at several levels (20, 30, 40 or 50% replacement), with appropriate essential amino acid (EAA) additions so that all
diets matched the EAA profiles of fish meal. In addition, we also included treatments in which those soybean replacement
diets were supplemented with either a) phytase, or b) taurine. All treatments were done in triplicate. The experiment began with
snakehead juveniles that averaged 4.7 g in size and lasted for 8 weeks. Growth of the fish on diets with 20% or 30% replacement
of fish meal with soybean meal was statistically indistinguishable from those fed the control fish meal (0% replacement)
diet, but diets with 40% or 50% fish meal replacement yielded statistically poorer growth. Addition of phytase or taurine to the
replacement diets enabled growth on the 40%, but not the 50%, replacement diet to be statistically indistinguishable from the
control. Thus, soybean meal can replace up to 30% of fish meal in the diet without addition of phytase or taurine, or 40% of
fish meal with the addition of phytase or taurine.
3

3
AMINO ACID SUPPLEMENTATION AND DIGESTIBILITY OF PRACTICAL ORGANIC
DIETS FOR NILE TILAPIA, Oreochromis niloticus, CONTAINING SOYBEAN MEAL AND
COMMERCIAL YEAST EXTRACT AS TOTAL REPLACEMENTS OF FISH MEAL
Kimberly A. Trosvik*, Linda S. Metts, Kenneth R. Thompson and Carl D. Webster
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
kimberly.trosvik@kysu.edu
Fish meal (FM) is considered the most nutritionally-complete protein source in diets for finfish due to its ideal balance of essential
fatty acids, amino acids, vitamins, and minerals. Fish meal’s nutritional profile is also desirable in the livestock and poultry
industries, which increases competition and demand for this resource, making fish meal the most expensive macro-ingredient
of a fish diet. There are numerous attractive protein sources that potentially could replace fish meal in aquaculture diets.
However, these protein sources often do not provide for similar growth in fish when compared to diets with fish meal. It has
been stated that the use of two or more complimentary protein sources may allow for growth results similar to ones found when
fish meal is added to a diet. Soybean meal and yeast are two favorable alternatives to fish meal that have shown some promise
when used in combination for fish meal replacement.
Tilapia are one of the most-cultured fin fish in the world. Organically-fed tilapia may allow producers to enter the rapidlydeveloping
organic sector of the market. The objective of this study was to assess the digestibility and effect of amino acid
supplementation in organic tilapia diets containing organically-certified yeast and soybean meal as a complete protein replacement
for fish meal.
A 6-week feeding trial was performed in a rack recirculating system comprised of 36, 10.0-L tanks stocked with 20, 200mg
fish per tank. Water temperature was maintained at 28 C. Fish were fed three times daily (0800, 1200 and 1600) all they could
consume in thirty minutes. Nine diets were formulated (Table 1) with 4 replicates per diet.
Data will be compared between final weight, feed conversion ratio, protein efficiency ratio, and specific growth rate. Results
will be discussed.
Table 1. Formulation of nine experimental diets fed to Nile tilapia.
Diet
1 2 3 4 5 6 7 8 9
Menhaden FM 20.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
NuPro© 0.0 0.0 0.0 15.0 30.0 45.0 15.0 30.0 45.0
Soybean meal 50.3 84.4 83.9 67.1 51.0 35.7 68.4 51.3 33.8
Lysine 0.0 0.0 0.3 0.0 0.0 0.0 0.4 0.4 0.4
Methionine 0.0 0.0 0.3 0.0 0.0 0.0 0.4 0.3 0.3
Other 29.7 15.6 15.5 17.9 19.0 19.3 15.8 18.0 20.5

SATURATED LIPID KEY TO MAXIMIZING FINISHING SUCCESS IN NILE TILAPIA
Oreochromis niloticus
Jesse Trushenski*, John Boesenberg and Christopher Kohler
Fisheries and Illinois Aquaculture Center
Southern Illinois University Carbondale
Carbondale, IL 62901-6511 USA
saluski@siu.edu
Fatty acid (FA) composition of cultured finfish can be tailored by transitioning fish reared on alternative lipid-based, low longchain
polyunsaturated fatty acids (LC-PUFA) content grow-out feeds to fish oil (FO)-based, high LC-PUFA ‘finishing’ feeds
prior to harvest. We have demonstrated the FA composition of the alternative lipid can affect overall finishing success and that
saturated lipid maximizes LC-PUFA augmentation during finishing of a lean-fleshed, cool water fish, sunshine bass. To determine
whether similar processes of lipid utilization and FA turnover occur in a lean-fleshed, warm-water species, coconut (CO),
grapeseed (GO), linseed (LO), and poultry (PO) oils were evaluated in Nile Tilapia grow-out feeds with respect to production
performance and responsiveness to finishing. Similar to sunshine bass, production performance of Nile Tilapia was unaffected
by application of the various feeding regimes, indicating this warm-water species can effectively utilize CO, GO, LO, PO, and
FO in aquafeeds. Implementation of the alternative lipid sources was associated with altered fillet FA composition, particularly
the accumulation of monounsaturated FA (MUFA) and medium-chain PUFA (MC-PUFA). Departures from a FO-associated
profile were largely corrected by finishing, but profile restoration was most complete among fish fed the CO– and PO-based
feeds during grow-out. These results suggest FA turnover in Nile Tilapia is generally similar to that of sunshine bass. However,
considerable retention of FO-associated FA was observed among Nile Tilapia fed alternative lipids, suggesting Nile Tilapia are
better able to conserve or synthesize LC-PUFA than sunshine bass.
3

3 0
SATURATED DIETARY LIPID SOURCES IN GROWOUT FEEDS FOR HYBRID STRIPED
BASS Morone chrysops ♀ x M. saxatilis ♂: ProDUction PerformAnce AnD fiLLet
FATTY ACID COMPOSITION
Jesse T. Trushenski*, John Boesenberg and Christopher C. Kohler
Fisheries and Illinois Aquaculture Center
Southern Illinois University Carbondale
Carbondale, IL 62901-6511 USA
saluski@siu.edu
Hybrid striped bass (HSB) generally tolerate
a wide range of alternative lipids in grow-out
feeds. However, grow-out feed fatty acid (FA)
composition influences long-chain polyunsaturated
FA (LC-PUFA) restoration during finishing.
Specifically, we have observed grow-out feeds
containing higher levels of saturated FA (SFA) to
yield fillets with greater LC-PUFA content after
finishing. To address whether different saturated
lipid sources are equivalent in this respect, we
fed juvenile (~25 g) HSB feeds (41/14% crude
protein/lipid) containing fish oil (100% FO),
coconut oil (100% CO), palm oil (100% PO), or
a 50:50 blend of FO and CO (50% CO) or PO
(50% PO). After 8 weeks of culture, production
performance was generally within acceptable
ranges for HSB; however, significantly reduced
feed intake and weight gain within the 100% CO
group suggest palatability and/or digestibility
of this feed may have been problematic (Table
1). Expectedly, fillet FA composition reflected
nutritional history (Figure 1): fillet levels of FOassociated
FA, particularly 20:5n-3 and 22:6n-3,
were reduced among fish fed the CO- and PObased
feeds. Increasing dietary inclusion of CO
and PO was associated with increased deposition
of SFA and MUFA, respectively, within the fillet.
Although fillet FA profile was significantly
altered in all CO- and PO-fed groups, LC-PUFA
content was better maintained among fish fed the
CO-based feeds. Although differences in fillet
LC-PUFA content between the CO- and PO-fed
fish were statistically significant, the magnitude
of these differences was relatively small.
Additional research is needed to determine
whether these differences in fillet profile would
affect overall finishing success. Assuming
issues of feed acceptance and/or utilization can
be rectified, CO and PO appear to be suitable
lipid sources for HSB grow-out feeds.

CHANGES IN WHOLE-BODY FATTY ACID CONCENTRATIONS IN FIRST-FEEDING
STEELHEAD Oncorhynchus mykiss FED DIETS CONTAINING PLANT OIL OR MARINE
FISH OIL
Ronald G. Twibell, Ann L. Gannam, Susan L. Ostrand, John S. A. Holmes and Jeff B. Poole
U. S. Fish and Wildlife Service
Abernathy Fish Technology Center
1440 Abernathy Creek Road
Longview, WA 98632 USA
Ronald_twibell@fws.gov
A 12-wk feeding trial was conducted to evaluate the effects of dietary lipids on growth response, carcass proximate composition
and carcass fatty acid concentrations in first-feeding steelhead. The two practical diets included a standard fish meal-based diet
top-coated with marine fish oil (pollock) and a marine product-free diet top-coated with vegetable oil (canola and flaxseed oils).
Each dietary treatment was fed to five replicate groups of 405 randomly selected first-feeding fry reared in fiberglass tanks.
Mean initial weight of the fish was 0.22 g/fish. Fish were collected every two weeks for carcass fatty acid analysis. At the end
of the study, fish fed the diet containing marine fish oil exhibited significantly higher (P

3 2
SUPPLEMENTAL NUTRITION PROVIDED TO PACIFIC WHITE SHRIMP Litopenaeus
vannamei BY NATURAL PRODUCTIVITY FROM EITHER A PHOTOAUTOTROPHIC OR A
HETEROTROPHIC HYPER-INTENSIVE ZERO-EXCHANGE BIOFLOC SYSTEM
Jesus A. Venero*, John Leffler, A.O. Galvez, Jason Haveman, Alisha Lawson, Andrew Shuler, Beth Thomas,
L. Vinatea and Craig L. Browdy
SCDNR - Waddell Mariculture Center
211 Sawmill Creek Road
Bluffton, SC 29910 USA
VeneroJ@dnr.sc.gov
Hyper-intensive zero-water exchange biofloc systems technologies are being developed to make the USA shrimp industry more
competitive. These systems support a diverse microbial community that has been shown to enhance shrimp growth and to
provide essential nutritional components. The nature of these enhancement factors is still uncertain, although a direct association
with marine microalgae, especially chlorophytes and diatoms, is suspected. A study was conducted in order to evaluate
the supplemental effect of natural productivity on Pacific white shrimp, L. vannamei, nutrition in zero-water exchange, high
intensity shrimp raceways. Two 50-m 2 greenhouse enclosed raceways were stocked with juvenile shrimp at 100/m 2 . One
raceway received direct sunlight leading to considerable photoautotrophic activity and the other was completely covered with
black polyethylene, blocking sunlight and creating a heterotrophic biofloc system. Water from each raceway was pumped
continuously through two different rows of sixteen 60-L glass tanks each located in an adjacent greenhouse. A third row of sixteen
glass tanks operating on a filtered UV-treated seawater recirculating system served as controls. Shrimp within each water
system were assigned to one of four 36%-crude protein diets: a fishmeal-based diet, an all-plant diet with DHA-AA and vitamin
supplementation, an all-plant diet without DHA-AA supplementation, and an all-plant diet without vitamin supplementation.
Each tank was stocked with 15 shrimp (0.79±0.03 g).
ANOVA analyses demonstrated no significant differences (P>0.05) among final weights or feed conversion ratios (FCR) for
shrimp raised in the three water systems when the DHA-AA deficient diet was fed. However, the same variables were significantly
improved (P≤0.05) for shrimp raised in the heterotrophic system than for shrimp raised in the other systems when the
other diets were fed (0.81 g/wk vs. 0.57-0.59 g/wk and 1.36 vs. 1.89-1.93, for weight gain/week and FCR respectively). Shrimp
consumed less than 50% of the total daily ration for all the diets, possibly due to poor palatability caused by a bad ingredient.
Apparently shrimp could not compensate for a dietary deficiency of DHA-AA by consuming biofloc when the food allowance
was reduced. However the heterotrophic biofloc apparently negated ill effects from the vitamin deficient diet. Better
growth performance and FCR ratios were observed when complete diets were fed in a heterotrophic system. Shrimp raised
in the autotrophic system had values comparable with shrimp raised in the clear water system, both significantly poorer than
those of the heterotrophic water. Currently this experiment is being repeated under similar conditions with enhancement of the
photoautotrophic system through periodic inoculation with microalgae of the genus Thallasiosira sp (pelagic) and Navicula sp
(bentonic).

EYE COLOR AS A PREDICTOR OF SOCIAL DOMINANCE IN NILE TILAPIA Oreochromis
niloticus
Emmanuel M. Vera Cruz*, Reggie May L. Bero, Remedios B. Bolivar and Russell J. Borski
Freshwater Aquaculture Center – College of Fisheries
Central Luzon State University
3120 Science City of Muñoz
Nueva Ecija, Philippines
bongbee15@hotmail.com
Heterogeneous fish growth is a common phenomenon in cultured fish populations. Variation in individual growth rates may
be attributed in part to social interactions and the formation of feeding hierarchies. Breeding is also largely driven by social behavior
and an understanding or ability to predict dominance may contribute to breeding programs. Fish in a stable dominance
hierarchy may be classified as dominant, subordinate, or intermediate in status.
It has previously been shown that eye color pattern in Nile tilapia is a consequence of a fish’s relative position in the social
hierarchy. However it is still an open question if this physiological difference can also be a cause of social status. To evaluate
this, 40 all-male juvenile Oreochromis niloticus of similar size and with no apparent differences in social history were isolated
for 10 days and were used in a social pair study. Eye color pattern of each fish was observed and recorded several times during
the isolation period, during an aggressive encounter and after the social interaction settled. Eye color change was marked by
fractional changes of the color of the iris and sclera around the pupil which was transformed into scores ranging from 0/8 (no
darkening) to 8/8 (total darkening).
Results of the study demonstrated that the ability to win a fight for social dominance can be predicted using the fish’s eye color.
Fish with darker eye color pattern are most likely to win the contest for social dominance. In addition, during an aggressive
encounter, shifts in the eye color pattern were observed in both the subsequent dominant and subordinate fish. This change in
eye color patterns in both fish served as a signal to the opponent to indicate preparedness to fight. When the social interactions
had settled, dominant and subordinate fish shifted their eye color patterns to a paler and darker configuration, respectively. The
eye darkening in the subordinates served as a social signal for social submission.
3 3

3 4
DIETARY CARBOHYDRATE LEVEL AFFECTS THE EXPRESSION OF GENES
REGULATING THE PRODUCTION OF REDUCTIVE POWER IN RAINBOW TROUT
Oncorhynchus mykiss, LIVER
Jurij Wacyk*, Madison Powell, Kenneth Rodnick, Gordon Murdoch, Barrie Robison, Rod Hill and Ronald Hardy
Hagerman Fish Culture Experimental Station (HFCES)
University of Idaho, Aquaculture Research Institute
3059F National Fish Hatchery Road
Hagerman, Idaho, 83332 USA
jwacyk@vandals.uidaho.edu.
The presence of carbohydrates (CHO) in plant ingredients and the fact that carnivorous fish like rainbow trout (RBT) are recognized
by their poor ability to use dietary carbohydrates represent a significant challenge for fish nutritionists. Considering that
high levels of dietary carbohydrates have been shown to be detrimental to fish performance, a study was designed to investigate
the effect of long-term dietary modifications (CHO levels) on the expression of genes related with the generation of reductive
power in trout liver. Seventy five RBT, average weight 28.3g, were distributed among 12 tanks (150 L), supplied with 6 L/min
(14.5ºC), spring water at the HFCES. Dietary treatments were formulated to be iso-nitrogenous (38% digestible protein) and
iso-lipidic (15% digestible lipid) with increasing levels of energy provided by gelatinized wheat starch. Fish were fed to apparent
satiation 3 times daily, 6 days a week for 12 weeks. Fish in each tank were bulk-weighed and counted at the beginning and
at the end of weeks 4 (T1), 8 (T2) and 12 (T3). At the same time periods, blood and liver samples were taken to monitor for
plasma glucose levels as well as gene expression changes. In our study we were successful in identifying significant differences
in plasma glucose and gene expression levels in the liver of rainbow trout. Increasing dietary starch level led to elevated levels
of plasma glucose. The response of this classic physiological parameter was expected and is characteristic of RBT fed diets
high in CHO. Comparing physiological and molecular data, it appears that the transcriptional machinery behind Glucose 6P
dehydrogenase expression, the regulatory entrance point to the pentose phosphate pathway, is able to adapt and closely follow
plasma glucose variations. The general up-regulation of Thioredoxin and Thioredoxin interacting protein by fish fed the wheat
starch containing diets may be indicative of an increased necessity for reductive power possibly linked with a modification of
the redox environment in the hepatocytes (REDOX stress).

INCREASING LEVELS OF DIETARY CARBOHYDRATES ARE ABLE TO
DIFFERENTIALLY MODIFY THE EXPRESSION LEVELS OF TRANSAMINASES IN
RAINBOW TROUT Oncorhynchus mykiss, LIVER
Jurij Wacyk*, Madison Powell, Kenneth Rodnick, Gordon Murdoch, Barrie Robison, Rod Hill and Ronald Hardy
Hagerman Fish Culture Experimental Station (HFCES)
University of Idaho, Aquaculture Research Institute
3059F National Fish Hatchery Road
Hagerman, Idaho, 83332 USA
jwacyk@vandals.uidaho.edu
Despite the fact that carnivorous fish are recognized by their poor ability to utilize dietary carbohydrates (CHO), the impact of
this group of nutrients in fish metabolism is far from being small and simple. Depending on factors like the CHO form and level
of inclusion among many others, fish like rainbow trout (RBT) are able to use them and improve productive performance.
One of the assumed advantages of using CHO as source of metabolic energy is spare dietary protein. As in mammals, fish
amino-transferases are key metabolic enzymes that remove amine of amino acids to free the carbon skeletons for purposes
like supplying energy. To gain further understanding on the protein sparing effect of dietary CHO a study was carried out to
evaluate changes in expression levels of alanine amino transferase (ALAT), aspartate amino transferase (ASAT) and glutamate
dehydrogenase (GlDH) in RBT. Seventy five RBT, with an average weight 28.3g, were distributed in each of 12 tanks (150
L) supplied with 6 L/min (14.5ºC), spring water at the HFCES, University of Idaho. Dietary treatments were formulated to
be iso-nitrogenous (38% digestible protein) and iso-lipidic (15% digestible lipid) with increasing levels of energy provided
by gelatinized wheat starch. Fish were fed to apparent satiation 3 times daily, 6 days a week for 12 weeks. Fish in each tank
were bulk-weighed and counted at the beginning and at the end of weeks 4, 8 and 12. At the same time periods, liver samples
were taken to monitor for liver glycogen levels as well as gene expression changes. Fish fed diets 0% and 15% CHO presented
significantly higher protein retention efficiencies than the other two groups. Increasing CHO levels in the diet generated a
significant increase in liver glycogen which was strongly and negatively correlated with ALAT expression levels (-0.75). The
other two transaminases did not follow a pattern of expression similar to ALAT even when significantly affected by the diet.
3

3
PLAYING WITH FOOD: EXAMINING REARING DIETS FOR WINTER FLOUNDER
Pseudopleuronectes americanus STOCK ENHANCEMENT THAT OPTIMIZE WEANING
SUCCESS IN THE HATCHERY AND IN THE WILD Pseudopleuronectes americanus STOCK
ENHANCEMENT THAT OPTIMIZE WEANING SUCCESS IN THE HATCHERY AND IN THE
WILD
Michelle L. Walsh*, Elizabeth A. Fairchild, Nathan Rennels, Stacy C. Farina, W. Huntting Howell,
Renee Mercaldo-Allen and Catherine Kuropat
University of New Hampshire
Durham, NH 03824 USA
michelle.walsh@unh.edu
Winter flounder (Pseudopleuronectes americanus) are capable of releasing hundreds of thousands of eggs annually but because
of the vulnerability of the small, early life-history stages, there is high natural mortality, and few survive to maturity. Captively
spawning, rearing, and releasing animals at a size or age beyond this mortality window may enhance natural stocks. A major
challenge of any captive rearing program, whether for aquaculture or stock enhancement, is providing the appropriate diet regimes
during development. Typically marine fish larvae are initially fed live food (e.g. rotifers, Artemia), and then weaned onto
formulated diets as they attain a size or developmental state that supports consumption of these artificial feeds. Weaning occurs
twice for fish that are used for stock enhancement; the second time occurs as they transition from the hatchery feed to natural
live diets once released. This research, which is part of a larger study designed to assess the feasibility of winter flounder stock
enhancement in New Hampshire, aims to identify rearing diets that optimize weaning success in the hatchery and minimize the
effects of subsequent weaning in the wild.
In 4-wk laboratory feeding trials, we analyzed the weaning
success of hatchery-reared, juvenile winter flounder raised
on different diets (both live and formulated) by quantifying
growth, survival, and RNA/DNA condition. Live diets included
late-stage nauplii of brine shrimp (Artemia franciscana), white
worms (Enchytraeus albidus) and burying marine amphipods
(Leptochierus plumulosus). Initial results indicate that fish
reared on live diets exhibit significantly higher growth (Fig. 1)
and survival than those reared on formulated diets (p < 0.05).
RNA/DNA condition was also higher in fish fed live feeds.
Subsequently, we examined the weaning success of these hatchery-reared
fish once released in the wild. In addition to growth
and survival, we determined the onset of feeding post-release,
gut fullness, and gut composition by performing controlled
cage
releases. Fish raised on live diets exhibited higher growth in
the wild than pellet-reared fish. This research provides information
that may promote advances in weaning strategies for stock
enhancement.

DEVELOPING VALUE-ADDED PRODUCTS FROM PADDLEFISH MEAT
Changzheng Wang*, Lingyu Huang, Cecil Butler, Richard J. Onders and Steven D. Mims
Human Nutrition Program
Kentucky State University
Frankfort, KY 40601 USA
changzheng.wang@kysu.edu
Hot smoke processing enhances the value of paddlefish meat. A group of experienced chefs indicated that whole paddlefish after
heading and gutting known as a bullet can be processed into a product that not only looks and tastes good, but also provides
the versatility chefs need to serve it in ways appealing to customers. The objective was to determine the optimal brine strength
and brining time needed to assure the adequate salt content (approx. 3.5%) of smoked whole paddlefish. Four paddlefish each
were brined either in 10% or 15% salt solution for either 24 hr or 48 hr. At the end of the brining, fish were rinsed in tap water
and left to dry at 4 o C overnight. They were hot smoked until the internal temperature reached 165 o C for 30 min. After cooling
down in a refrigerator, the smoked fish were vacuum-packed and stored at -20C before analysis. The smoked meat was
homogenized in a grinder. Two gram samples were soaked in distilled water for two hrs. The supernatant was used for salt
analysis by a salt analyzer. Higher brine strength and longer brining time resulted in significantly higher salt content in the
smoked fish. Fish brined for 24 hr in the 10% brine achieved adequate level of salt content without making the product overly
salty. The results indicate that 10% salt solution and 24 hr brining time should be used to brine whole paddlefish before smoking
processing for a consumer-safe product.
3

3
EVALUATION OF PREPARATION PROCEDURES FOR PADDLEFISH CAVIAR
Changzheng Wang, Steven D. Mims, Lingyu Huang and Richard J. Onders
Human Nutrition Program
Aquaculture Research Center
Kentucky State University
Frankfort, KY 40601 USA
Caviars are in high demand in the market place because of the supply from Russian sturgeon is almost depleted. The potential is
great for paddlefish eggs to be made into caviars for the benefits of both the consumers and the producers. Various procedures
for caviar preparation have been used and their effects on product quality are not well defined. The objective of this study was
to determine the effects of salt concentration and washing the eggs on the quality of paddlefish caviar. Eggs from two mature
paddlefish were hand-pressed through a stainless steel screen so that the eggs were separated from each other. The connective
tissues were discarded. The eggs were divided into batches of 200 g placed in clear plastic containers. Half of the batches were
not washed, whereas the other half was washed with tap water before salt was added into the eggs. The amount of salt added
was 2.5%. 3.5% or 4.5% of the egg weight. The salt was mixed into the eggs evenly before the containers were covered and
placed at 4 C. The color, texture and salt content were determined within a week. A taste panel of chefs experienced with caviar
evaluated the products. The water-phase salt content was above 3.5% for caviars prepared with the lowest level of salt added
(2.5%). Higher amount of salt resulted in much higher salt content in the products and a saltier taste. However, the visual and
other taste profiles of the products were not significantly affected by the preparation procedures. Therefore, washing the eggs
is optional for caviar preparation. Adding salts above 2.5% of the egg weight will result in salty caviar.

PROGRESS AND CONSTRAINTS TOWARD COMMERCIALIZATION OF BLACK SEA BASS
AQUACULTURE IN THE US: HATCHERY, NURSERY, GROWOUT AND MARKETING
W. O. Watanabe*, M. S. Alam, C. F. Dumas, R. F. Lee, T. M. Losordo, D. DeLong, D. Berlinsky, G. Nardi,
C. D. Bentley, T. C. Rezek, K. Sullivan, J. Wilde and A. Myers
University North Carolina Wilmington (UNCW)
Center for Marine Science
601 S. College Rd.
Wilmington, NC 28403-5927 USA
watanabew@uncw.edu
Black sea bass (BSB) Centropristis striata is a heavily exploited marine finfish inhabiting continental shelf waters of the eastern
US and the subject of state and federally supported aquaculture research. Recent progress in research and pilot commercial
BSB aquaculture are reviewed. Protocols for hormone-induced spawning have been improved by optimizing dose of hormone
administration to maximize egg and larval quality. Suitable environmental conditions and feeding regimens for larval culture
have been delineated. In 2007, approximately 50,000 juveniles were produced in pilot-scale trials at a commercial hatchery in
NH (Great Bay Aquaculture). Optimal protein level for growth of juvenile BSB fed a menhaden fish meal based diet is 44%,
but a high percentage (60-70%) of soybean meal has been effectively substituted for fish meal, indicating an excellent potential
for formulating sustainable practical feeds. Disparate rearing conditions have been used in independent studies on growout of
hatchery-reared fingerlings to marketable stages, and significant variability in growth rates have been observed. In NC, BSB
fingerlings (N = 3,300, mean wt. = 27 g, age = 125 d post-hatching) were stocked in a recirculating aquaculture system (RAS)
consisting of two 16-m 3 tanks and reared at an average of 21.0 o C and 33 g/L. Fish were fed a commercial diet (55% P, 18% L)
and reached an average premium size of 682 g (range = 328-1,350 g) in 570 d (age = 695 dph) with 77% survival and a final
biomass density of 52 kg/m 3 . In GA, however, BSB (N = 200, mean wt. = 100 g, age = 294 d) fed a diet of live juvenile tilapia
in a RAS at an average of 24 o C reached a mean of 900 g in only 274 d (age = 568 dph), with growth presumably increased by
diet and temperature. A prototype, closed (zero-discharge) marine RAS was tested for BSB growout in Raleigh, 150 mi from
the ocean. In its 12th month of completely closed operation, the water renovation system, treating 24 g/L artificial seawater,
has evolved to meet the challenges of a growing biomass of BSB. In BSB, protogynous hermaphroditism can be disrupted in
culture, with individuals differentiating first as males showing faster growth. High variation in individual body weights among
fish has been observed during BSB growout, suggesting that early culling of slow-growing fish will reduce growout time under
practical culture. Pasteurellosis (Photobacterium damselae piscicida) was infrequently observed in subadult fish raised in RAS
in NC and was remedied by lowering water temperature to 20-21 o C. Although etiology appeared non-pathogenic, exopthalmia
(“popeye”) has been a recurring disorder in BSB raised in intensive RAS, but causing minor losses to date. Based on comprehensive
marketing studies, current annual niche market demand for fresh BSB (whole on ice @ $7.50 per lb) in the state
of NC is estimated at 310,977 lbs. Out-of-state metro markets are under study and should boost demand. Pilot commercial
project for BSB are underway in NC and in VA. Improving cost and availability of fingerlings, growth rates (via domestication
and sex control), zero-exchange RAS technology, and development of sustainable feeds are keys to the transition from pilot to
commercial scale production.
3

3 0
NEW TECHNIQUE FOR ADMINISTRATION OF HUMAN CHORIONIC GONADOTROPIN
– “OVARIAN LAVAGE”
Craig Watson*, Amy Wood, and J. Scott Graves
Tropical Aquaculture Laboratory
University of Florida
1408 24 th Street S.E.
Ruskin, FL 33570 USA
For a number of varying factors, certain species of fish do not respond to standard injection methods for administration of
spawning hormones. Research with two new species of fish, Tetraodon nigriviridis and Mastacembelus erythrotaenia, resulted
in the development of an alternative method for application of Chorulon®, via direct lavage into the ovaries. This presentation
will discuss the issues which led to this development, as well as the dosage and application details.
CAPTIVE REPRODUCTION OF Tetraodon nigriviridis; SERENDIPITY IN AQUACULTURE
Craig Watson, Scott Graves, Kathy Heym, Jeff Hill and Amy Wood
Tropical Aquaculture Laboratory
University of Florida
1408 24 th Street S.E.
Ruskin, FL 33570 USA
cawatson@ufl.edu
Tetraodon nigriviridis, or the Green Spotted Puffer, is a commonly sold aquarium fish collected from rivers and estuaries of
Asia. It was listed as a priority species that would be profitable for Florida farms if production techniques were developed.
Early on in our research, a literature review and internet inquiries led to the discovery that this species also is an extremely
important animal to genomic studies, having the shortest known genome of any invertebrate species. Scientists studying this
species were also limited to wild-collected juveniles and adults, which greatly limited the types of studies which could occur.
Our successful reproduction strategies will shortly make this species available to the hobby, but also promise to lead to new
discoveries as we can now produce embryos. Publications in progress include a detailed discussion of the 4 day embryology
as well. This development will provide an additional, totally unexpected market for Florida farms, providing researchers with
captive bred fish of a known lineage, fertilized embryos of varying age, and potentially transgenic fish.

EFFECT OF CULTURE DENSITY ON PRODUCTION CHARACTERISTICS AND
BODY COMPOSITION OF MARKET SIZE COBIA REARED FROM JUVENILES IN
RECIRCULATING AQUACULTURE SYSTEMS
Charles R. Weirich*, Paul S. Wills, Richard M. Baptiste and Marty A. Riche
USDA Agricultural Research Service and Center for Aquaculture and Stock Enhancement
Harbor Branch Oceanographic Institute at Florida Atlantic University
5600 US Highway 1 North
Ft. Pierce, FL 34946 USA
charles.weirich@ars.usda.gov
Interest regarding cobia Rachycentron canadum aquaculture in the US has increased greatly in the last decade due to their
excellent consumer appeal, extremely rapid growth rates, and the observed success of rearing this species in Taiwan and other
southeastern Asian countries. Because the principal culture system currently employed for farming this species is offshore
net pens or cages, only limited information exists with respect to rearing juvenile cobia to marketable sizes using recirculating
aquaculture systems (RAS).
A 119-day trial to compare three rearing densities on production characteristics and body composition of sub-adult cobia was
completed using four replicate 45-m 3 RAS, each containing three experimental 8-m 3 culture tanks (one tank/density/system)
located within the USDA-ARS Sustainable Tank Aquaculture Recirculating Research (STARR) facility on the campus of Harbor
Branch Oceanographic Institute at Florida Atlantic University. To initiate the trial juvenile cobia (initial weight = 322 g)
were stocked into three tanks of each RAS, each at a different density: low (1.4 kg/m 3 , 35 fish); medium (2.8 kg/m 3 , 70 fish);
or high (4.2 kg/m 3 , 106 fish). After stocking, fish were fed a commercial slow sinking pelleted diet (45% CP, 15% CL) twice
daily at a targeted daily ration of 3 to 5 % bw/d. Initial pellet size was 5 mm and was increased as fish grew. At three week
intervals 10% of the population in each tank was sampled to estimate mean weight, weight gain, specific growth rate (SGR),
feed conversion efficiency (FCE), biomass, and survival. At termination of the trial, tanks were clean harvested to determine
production characteristics and sufficient fish were processed to determine hepatosomatic index, % visceral fat, fillet yield,
and proximate composition indices. Although tissue samples have not yet been analyzed, results indicate that rearing density
had no effect on production throughout and at the conclusion of the trial. Fish reared at all three densities exhibited excellent
growth, feed conversion, and survival. Final biomass of low, medium, and high density treatments was 9.0, 18.1, and 27.8
kg/m 3 , respectively. Across treatment fillet yield was 42.1% and average total fillet weight of each harvested fish was 884 g.
Because no density effect was observed, higher densities are currently being evaluated. In addition, target harvest weight will
be increased to 3 kg.
Final mean weight, specific growth rate (SGR), feed conversion rate (FCR), and
survival of juvenile cobia (initial weight = 322 g) reared at three different densities
for 119 days. No significant differences existed between density treatments for
each variable.
Density Weight (kg) SGR (%/d) FCE (%) Survival (%)
Low
Medium
High
2.13
2.15
2.15
1.58
1.60
1.59
66.1
65.1
65.7
95.7
96.1
97.2
3 1

3 2
EVALUATION OF COMMERCIAL ARTEMIA ENRICHMENT PRODUCTS ON GROWTH,
SURVIVAL, STRESS TOLERANCE, AND FATTY ACID COMPOSITION OF LARVAL
FLORIDA POMPANO
Charles R. Weirich
USDA Agricultural Research Service
Harbor Branch Oceanographic Institute at Florida Atlantic University
5600 US Highway 1 North
Ft. Pierce, FL 34946 USA
charles.weirich@ars.usda.gov
Rearing live feed organisms represents the majority of time and labor associated with larviculture operations vital to the seed
stock supply of marine finfish aquaculture production facilities. No information exists with respect to optimal Artemia enrichment
protocols for Florida pompano Trachinotus carolinus, a high-value species with excellent potential for aquaculture in
the US. This study compared growth, survival, stress tolerance, and fatty acid composition of pompano larvae fed Artemia
enriched with selected commercial diets.
Two 9-day rearing trials were conducted. In Trial 1, the enrichment products DC DHA Selco (Selco), Ori-Green (OG), and
AlgaMac3050 (AM) were evaluated. In Trial 2, Selco, Ori-Pro (OP), and a developmental product (Selco QI) were compared.
In both trials, larvae were reared using protocols developed at our facility and at 10 days post-hatch (DPH) were stocked into a
recirculating aquaculture system composed of 12, 110-L tanks at a rate of 7.7 (Trial 1) and 9.1 larvae/L (Trial 2). Four replicate
tanks were used for each enrichment treatment. At 11 DPH, larvae were fed 2 nd instar Artemia enriched with experimental
diets four times daily at a rate of 2-2.5 prey/ml/feeding. At 20 DPH, wet and dry weights of recently metamorphosed juveniles
were determined, and total length (TL), standard length (SL), and body depth (BD) were measured by image analysis. Stress
tolerance was assessed by acute hypersaline exposure (100 g/L). Survival was also determined and Artemia and fish samples
were collected for fatty acid analysis.
Results revealed that with the exception of dry weight in Trial 1 and BD in Trial 2, no differences existed among enrichment
diets with respect to growth, as well as survival. In Trial 1, stress tolerance of fish fed Artemia enriched with AM was reduced
relative to that of fish fed Selco and OG-enriched Artemia. In Trial 2, fish fed Artemia enriched with OP died immediately after
being transferred to hypersaline conditions. Fatty acid composition of Artemia enriched with tested products will be presented
as well as that of sampled fish.
Length, weight, and survival of 20 DPH pompano fed Artemia enriched with
selected commercial diets for 9 d in Trials 1 and 2
TL SL BD Wet wt. Dry wt. Survival
Diet (mm) (mm) (mm) (mg) (mg) (%)
Trial 1
Selco 14.4 a 11.5 a 3.9 a 42.4 a 9.6 a 55.9 a
OG 14.1 a 11.2 a 3.8 a 41.0 a 8.0 b 57.6 a
AM 14.5 a 11.5 a 3.7 a 36.0 a 9.0 ab 62.1 a
Trial 2
Selco 11.0 a 8.7 a 2.9 b 19.8 a 4.7 a 84.0 a
OP 11.3 a 9.0 a 3.2 a 21.1 a 4.7 a 85.2 a
Selco QI 11.1 a 8.8 a 2.9 b 20.1 a 5.0 a 84.4 a

BAYESIAN METHODS FOR DETERMINING THE IMPORTANCE OF EFFECTSBAYESIAN
METHODS FOR DETERMINING THE IMPORTANCE OF EFFECTS
Tom L. Welker, Tim L. Welker and Phillip H. Klesius
USDA, ARS
Aquatic Animal Health Research Unit
990 Wire Road
Auburn, AL 36832 USA
thomas.welker@ars.usda.gov
Criticisms have plagued the frequentist null-hypothesis significance testing (NHST) procedure since the time day it was fashioned
created from the Fisher Significance Test and Hypothesis Test of Jerzy Neyman and Egon Pearson. Alternatives to NHST
exist in frequentist statistics, but competing methods are also available in Bayesian statistics, which have important advantages
over frequentist procedures. Bayesian methods, however, have been little used to determine the importance of effects, primarily
due to unfamiliarity stemming from the dominance of frequentist statistical procedures during the 20 th century, lack of practical
information on application of the methodologies, and criticisms of the use of posterior probabilities. However, rRecent methods
designed to help bridge the Bayesian – frequentist gap have been developed and overcome the perceived subjective bias of
Bayesian posterior probabilities through the use of non-informative priors. One such method integrates Bayes theorem within
a frequentist-type ANOVA framework familiar to most researchers. This Bayesian approach leads to conclusions of the importance
of effects based on probability and permits a reinterpretation of the usual confidence interval to determine differences
between means and assertions of largeness or smallness of effect, in which the latter has no frequentist counterpart. Examples of
a Bayesian alternative to ANOVA applied to aquaculture will be presented and compared to standard frequentist techniques.
3 3

3 4
ANALYSIS OF NICHE MARKET DEMAND FOR FARM-RAISED BLACK SEA BASS
Centropristis striata IN THREE MAJOR METROPOLITAN U.S. CITIES: ATLANTA,
PHILADELPHIA AND NEW YORK CITY
James D. Wilde*, Christopher F. Dumas and Wade O. Watanabe
Center for Marine Science Research
University of North Carolina Wilmington
10 South Cardinal Dr. Suite 2000
Wilmington, NC 28403
jdwilde@gmail.com
The black sea bass (BSB), Centropristis striata, is a marine finfish found naturally along the east coast of the United States.
Historically, BSB has been an important commercial seafood species with strong demand in major east coast U.S. cities.
However, overexploitation is resulting in restrictive regulations that will limit future commercial landings. BSB is a highvalue
fish, typically prepared fresh in upscale gourmet restaurants. High value and an increasingly limited wild-caught supply
make BSB an excellent candidate for commercial aquaculture. Due to the high capital costs of marine aquaculture production,
identifying high-value market niches is essential. At WAS 2008, the authors presented estimates of niche market demand for
BSB for the state of North Carolina, USA, a leading state in the development of BSB production technology. The purpose of
the present work is to estimate BSB niche market demand in large metropolitan markets along the east coast of the USA.
We investigate the high-value niche market for farm-raised BSB in three east coast USA metropolitan cities (Philadelphia,
New York City and Atlanta) using an in-restaurant, field sample survey of restaurant chefs and managers. Methods follow
those used successfully in North Carolina. BSB broodstock have been spawned and juveniles grown out to market size in a
near commercial-scale recirculating aquaculture system (RAS) located at the Center for Marine Science, University of North
Carolina Wilmington, USA. Approximately 1,700 RAS-grown BSB provide fresh product for the in-restaurant surveys, where
chefs will prepare the product and compare it with substitutes. We analyze restaurant and consumer preferences, investigating
the effects of restaurant seating capacity, average entree price, ethnic theme, tourist/local clientele, seafood intensity, purchase
frequency and storage duration, preferred product form and preparation method, BSB size and fat content, BSB price, substitute
fish type and price, and season on BSB quantity demanded (pounds per month) at the individual restaurant level. Our sampling
design allows us to identify and characterize the size of the BSB upscale niche market and to extrapolate sample results to the
full population of metropolitan restaurants to estimate the demand for niche market BSB in major east coast USA cities.

GROWTH PARAMETERS OF WILD AND SELECTED STRAINS OF ATLANTIC SALMON
(Salmo salar) ON TWO EXPERIMENTAL DIETS
William R. Wolters*, Frederic T. Barrows, Gary S. Burr and Ronald W. Hardy
USDA, ARS National Cold Water Marine Aquaculture Center
25 Salmon Farm Road
Franklin, ME 04634 USA
bill.wolters@ars.usda.gov
Two Atlantic salmon strains, one wild and one selected, were cultured from parr to smolt size to evaluate growth parameters,
feed consumption, and feed conversion. Each strain was fed diets formulated to represent either a traditional high protein, lower
energy salmon diet (46% protein, 18% fat) or a newer high energy diet (40% protein, 32% fat).
Atlantic salmon parr from Penobscot (wild) and St. John’s River (selected) strains were cultured in 265-liter tanks filled with
2-3 ppt salinity well water and connected to a common bio-filter system. Salmon parr were stocked at approximately 5 kg/m3
(100 fish/tank) and fed one of two experimental diets in a 2 x 2 factorial design with 4 replicate tanks for each treatment. Fish
were fed to apparent satiation with feed amounts adjusted weekly. Both diets were produced using twin-screw cooking extrusion,
and fish oil was vacuum coated after manufacture. Fish were anesthetized, counted and weighed at approximately 2 week
intervals. Analysis of variance was used to determine the effects of strain, diet, and diet × strain interaction on initial weight,
day 163 weight, specific growth rate, thermal growth coefficient, % feed consumed/day, and feed conversion ratio.
There were significant strain differences in initial weight, day 163 weight, specific growth rate, % feed consumed per day,
and thermal growth coefficient. Diet also had a significant effect on day 163 weight, specific growth rate, and thermal growth
coefficient. There were no differences from strain or diet for feed conversion or strain × diet interactions. St. John’s strain
salmon grew faster and were larger after 163 days (117.2+2.5g) than Penobscot River salmon (71.2+2.5g) (Figure 1). Although
diet had a significant effect, 163 day weight was different between the two experimental diets only for St. John’s River fish
(99.0+2.0 and 89.4+2.0g). Strain differences accounted for 85.9% of the difference in 163 day weight compared to 14.2% due
to diet. Results indicate St. John’s River salmon, which are commercially cultured and are being used in selective breeding
programs, have superior growth compared to wild, unselected Penobscot River salmon, and have even faster growth when fed
modern type diets.
3

3
MATERNAL LIPID NUTRITION AFFECTS ZEBRAFISH Danio rerio OOCYTE FATTY
ACID COMPOSITION AND FERTILIZATION SUCCESS BUT NOT LARVAL GROWTH OR
SURVIVAL
Jesse Trushenski, Dan Theisen and Curry Woods*
Department of Animal and Avian Sciences
University of Maryland
College Park, MD 20742 USA
Lipid nutrition can influence reproductive performance in finfish, as well as alter oocyte lipid composition. In turn, alterations
in oocyte fatty acid (FA) composition can affect gamete quality and larval vigor. Provision of long-chain, polyunsaturated FA
(LC-PUFA) has proven critical for ensuring reproductive and progeny success in several species. We assessed whether differences
in nutritional history, i.e. differential intake of LC-PUFA, among zebrafish broodstock, influence oocyte FA composition
and viability of the resultant progeny.
Adult zebrafish were stocked into 3, 10-L tanks (20 females + 10 males/tank) within a recirculation system. Each tank was fed
ad libidum for 4 weeks according to 1 of 3 experimental feeding regimens: feeding commercial feed only (“Z”; Complete Adult
Zebrafish feed, Zeigler Bros., Inc., Gardners, PA, USA), 2) commercial feed + live Artemia spp. (“Z+A”; San Francisco Bay
Brand, Aquatic Ecosystems, Inc., Apopka, FL, USA), or 3) commercial feed + freeze-dried copepods (“Z+C”; Cyclop-eeze®,
Argent Chemical Laboratories, Redmond, WA, USA) After completion of the conditioning period, oocytes were collected
weekly until 5 replicate spawns (N=5) of 100+ oocytes were recovered from each experimental group.
Oocyte FA profile reflected broodstock nutritional history. Oocytes from the Z and Z+C groups contain greater amounts of
LC-PUFA, whereas those from the Z+A group contained more monounsaturates (MUFA) and medium-chain polyunsaturates
(MC-PUFA). Hatch, larval survival and growth were equivalent among the experimental groups, however, fertilization
success was significantly greater among the Z and Z+C groups. Our data suggest oocyte LC-PUFA concentrations are related
to fertilization success in zebrafish, and failure to provide sufficient amounts of these FA in broodstock diets may impact larval
production.

QUALITY ASSESSMENT OF ATLANTIC STURGEON Acipenser oxyrinchus SPERMATOZOA
UNDER EXPERIMENTAL SHORT-TERM STORAGE CONDITIONS
L. C. Woods III*, Katy Dorsey, Dan Theisen, Frank Siewerdt, Jerre Mohler, Brian Richardson, Glenn Welsh,
Timothy Conn and David Guthrie
Department of Animal and Avian Sciences
University of Maryland
College Park, MD 20742 USA
There is significant interest to restore the Atlantic sturgeon, a species of concern. Biologists are interested in both the short-term
storage and cryopreservation of semen to maximize availability of viable spermatozoa whenever a rare ripe female is found
and available for spawning. We conducted short-term storage trials on semen obtained both from captive males, held at the
US Fish and Wildlife Service’s Fish Technology Center, Lamar, Pennsylvania that were hormonally induced to spermiate; and
wild males collected during the spawning season from the Hudson River this past year. Testes of all fish were catheterized to
collect semen. Semen samples with motility at the time of collection > 90% were used in initial experiments to quantitatively
examine cell quality over time under experimental conditions. Semen samples were stored under refrigeration (4 + 1oC) in two
experimental gas environments: oxygen or nitrogen. Samples in each gas environment were additionally subdivided into three
experimental dilutions: two experimental extenders and neat, or no dilution of the semen.
Semen samples were collected from individual males and transported chilled, at 4-6 o C, to the lab. Upon arrival, analyses of
gamete quality for each sample were performed prior to the administration of any experimental treatments. Approximately 24
hours post-arrival (Day 1) and then every other day for one week (i.e. Day 3, 5, and 7), experimental semen samples were again
quantitatively analyzed for quality. Sperm quality parameters evaluated included: motion analysis using a computer assisted
sperm analysis system, viability using a flow cytometer, and cellular ATP levels using a Luciferin-Luciferase bioluminescence
assay. Our results indicated that Atlantic sturgeon spermatozoa stored under oxygen retained higher quality than those stored
in the absence of oxygen. One experimental diluent, appears to mediate the harmful effects of a storage environment that is
without oxygen for up to one week.
3

3
EFFECTS OF POST-PROCESSING STORAGE TIME AND TEMPERATURE ON FISHMEAL
PRODUCED FROM PINK SALMON Oncorhynchus gorbusch BYPRODUCTS
Ted H. Wu* and Peter J. Bechtel
United States Department of Agriculture/Agricultural Research Service,
PO Box 757200
Fairbanks, AK 99775 USA
ted.wu@ars.usda.gov
Alaska’s wild commercial Pacific salmon harvest is over 400,000 metric tons annually. The processing of this catch resulted in
over 110,000 metric tons of byproducts available for production into fish meals and oils. When the initial raw material is not
directly processed the quality of the fish meal is affected. The objective of this study was to examine fish meal produced from
aging raw pink salmon byproducts stored at two temperatures.
Raw pink salmon byproducts were stored at 6 °C from 0-10 days and 15 °C from 0-4 days. Each day a portion of the raw
byproduct was processed into fish meal. Nutritional and chemical analyses were performed on the raw byproducts and
processed meals. The initial starting raw byproducts had values of total volatile base nitrogen (TVB-N) at 36 mg N/ 100g,
histamine at 4.2 mg/kg and thiobarbituric acid reactive substances (TBARS) at 1.9 mg malonaldehyde (MDA)/kg, respectively.
TVB-N values indicated substantial spoilage by day 2 at 15 °C and day 6 at 6 °C
(Fig 1.). Most biogenic amine concentrations of raw byproduct stored at 15 °C increased at day 1 and by day 2 were many
significant differences. There was little temperature dependence for lipid oxidation associated with the respective temperatures.
No great differences in fish meals processed from byproducts were noted in the amino acid profile with storage time or
temperature changes. Biogenic amines in fish meals suggested some changes occurring between day 0 and storage day 3 at
15 °C and storage day 6 at 6 °C. TBARS values in the fish meal at day 2 for both 15 °C and 6 °C showed significant changed
from the starting material.
While chemical indications suggested spoilage of the byproducts with increased storage and temperatures, a lower quality
meal with elevated biogenic amine values and TBARS can still be produced that has a good amino acid profile and essential
minerals.

SMALL-SCALE, LOW-INPUT AQUACULTURE: RETURN ON AN INVESTMENT
William A. Wurts
Kentucky State University Cooperative Extension Program
P.O. Box 469
1205 Hopkinsville Street
UK Research and Education Center
Princeton, KY 42445-0469 USA
www.ca.uky.edu/wkrec/Wurtspage.htm
Aquaculture is typically practiced intensively on large scales. First year start-up and production costs (per acre) for intensive
aquaculture can be prohibitive. Aerators, pond-side electricity, substrate, and high stocking densities are used to raise freshwater
shrimp intensively. Basic start-up and initial production costs for the first year, excluding land purchase and pond construction,
can be higher than $8,000/ac. These costs can be reduced by as much as 73 % with low-input shrimp farming practices.
While somewhat less expensive for intensive catfish farming, initial outlays could easily reach several thousand dollars per
acre. Again, expenditures can be significantly lowered by reducing inputs.
In addition to higher costs, intensive aquaculture practices generate higher yields. This leads to the need for wholesale markets.
Wholesale markets pay lower prices. Therefore, to make significant income, many acres of ponds are needed. Overall, this
makes commercial aquaculture impractical for many land-owners and small farmers with limited acreage and capital.
Alternatively, small-scale, low-input aquaculture can be an attractive “investment” for those with a few thousand dollars and an
acre or two to work with. Reduced inputs and acreage minimize start-up costs, but per acre yield will be lower too. However,
this makes niche marketing and retail sales more manageable and viable. Retail sales command higher prices. Combining the
lower costs of reduced input production with higher prices from retail sales can provide substantially greater profits per acre
than can be achieved with the same limited acreage under intensive production. For example, let us assume an initial investment
of $10,000 for one, 2-acre production pond. Annual profit for intensive production will be $200/acre at wholesale prices
and $1,000/acre for low-input production at retail prices. Therefore, the net annual return on the investment is 2% for the
intensive practice and 10% for the low-input practice. In other words, low-input practices might provide a substantially greater
annual return on the initial $10,000 investment. Furthermore, the potential risks and losses associated with low input practices
are much lower. While small-scale, low-input production may not provide enough income to live on, it may offer an attractive
opportunity for a high rate of return with a limited capital investment.
3

3 0
AQUATIC PLANT AND ALGAE CONTROL TRAINING FOR COMMERCIAL AND PRIVATE
PESTICIDE APPLICATORS IN KENTUCKY
Forrest Wynne
Kentucky State University Cooperative Extension Program
Graves County Extension Office
251 Housman Street
Mayfield, KY 42066 USA
fwynne@email.uky.edu
Aquatic plant and algae control training sessions were offered to commercial and private pesticide applicators during the winter
and spring of 2007-2008. Commercial and private applicators that use restricted pesticides must be certified by the Kentucky
Department of Agricultures’ (KDA) Division of Environmental Services. A three year certification is granted upon passing
written exams for private and commercial applicators. Before these trainings were offered, there were no KDA approved programs
in Kentucky that would allow Continuing Education Units (CEU’s) towards re-certification for commercial applicators
of aquatic pesticides. Neighboring states offered little reciprocity between the few agency programs available. Commercial
applicators traveled out of the region to attend training programs and get the CEU’s they needed for recertification or would
re-take the certification test before their license expired.
Training sessions were conducted at three locations during the winter and spring of 2007-2008. Various agency personnel, retail
herbicide distributors and commercial and private applicators attended the sessions. The sessions were advertised on the KDA
Division of Environmental Services website listing continuing education training programs and on the University of Kentucky
College of Agriculture website listing in-service training programs for cooperative extension employees. Three Kentucky State
University State Extension Specialists for aquaculture conducted the half day sessions at university facilities. The programs
emphasized proper plant identification, water quality management and algae control, pond construction and the management of
recreational ponds, and the biological, mechanical, physical and chemical methods of controlling aquatic vegetation. The use
of available resource materials were emphasized including aquatic plant identification and herbicide information websites, fact
sheets, printed material, identification guides, and the appropriate state agency personnel to contact for further assistance.

IMMUNIZATION OF CATFISH WITH INACTIVATED TROPHONTS AGAINST
Ichthyophthirius
De-Hai Xu*, Phillip H. Klesius and Craig A. Shoemaker
USDA, Agricultural Research Service
Aquatic Animal Health Research Laboratory
990 Wire Road
Auburn, AL 36832 USA
dehai.xu@ars.usda.gov
The protozoan parasite Ichthyophthirius multifiliis (Ich) infests most species of fresh water fish worldwide and damages fish
skin and gills. Epizootics have been reported in various freshwater fishes worldwide and result in severe economic loss to aquaculture.
There are no chemical treatments effective in controlling Ich infection. Vaccination offers a preventive method against
Ich. Various results have been reported in fish immunized with different Ich trophont preparations. The factors that influenced
immunity following trophont immunization are not clear and need further study. Two trials were conducted to determine the
effect of immunization with inactivated trophonts on serum and cutaneous antibody titers and survival in channel catfish.
In trial I, catfish were intraperitoneally (IP) immunized with: 1) 1% formalin-inactivated trophonts, 2) 3% formalin-inactivated
trophonts and 3) freeze-thawed trophonts. Positive and negative control catfish were immunized with live theronts and 5% bovine
serum albumin (BSA), respectively. The formalin-inactivated or frozen trophonts were separately sonicated and adjusted
to 2000 trophonts/ml in PBS for vaccination. Each fish was IP injected 10 µl/g fish of the antigen suspension corresponding to
20 trophonts/g fish. At day 14, 28 and 50 post immunizations, no statistical difference was noted in anti-Ich antibody titers
in fish immunized with formalin-inactivated trophonts or freeze-thawed trophonts. The survival of immunized catfish ranged
from 33.3% - 43.3% for the formalin-inactivated or freeze-thawed trophonts at 50 d post-immunization after challenge with
theronts at a dose of 15,000 theronts per fish. The survival of the live theront and BSA-immunized catfish was 93.3 and 0%,
respectively.
In trial II, catfish were IP-immunized with sonicated trophonts at doses of 1) 5 trophonts/g fish, 2) 10 trophonts/g fish, 3) 20
trophonts/g fish and 4) 5% BSA as the control. Fish immunized with 10 or 20 trophonts/g fish showed highest serum (1/210-
1/480) and cutaneous antibody titers (1/48-1/52), respectively at 22 d post-immunization and survival (63.3 -60.0%). The fish
immunized with 5 trophonts/g fish had titers of 1/52 and 1/12 for serum and cutaneous antibody and survival of 23.3%. BSA
immunized catfish had background titers and a survival of 6.7%. There was a significant correlation between dose of sonicated
trophonts used to immunize the fish and catfish survival (correlation coefficient=0.859, p

3 2
SPERM CRYOPRESERVATION IN BIOMEDICAL RESEARCH FISH MODELS
Huiping Yang* and Terrence R. Tiersch
Aquaculture Research Station
Louisiana State University Agricultural Center
2410 Ben Hur Road
Baton Rouge, Louisiana 70820 USA
hyang@agcenter.lsu.edu
There is increasing interest in sperm cryopreservation for fish models used in biomedical research. Basic biological characteristics
regarding natural habitat, testis structure, spermatogenesis, sperm morphology, and sperm physiology have been studied
in these fishes. Protocols for sperm cryopreservation have been developed and require comparison and standardization (Table
1). Evaluation of the viability of thawed sperm by artificial fertilization should be emphasized. Future research should focus
on standardization of procedures, gamete quality evaluation, high-throughput processing, and establishment of working repositories.
Table 1. Comparison of sperm cryopreservation in zebrafish, medaka, and Xiphophorus fishes
Extender Cryoprotectant Packaging Cooling rate Thawing method Fertilization
Zebrafish (Danio rerio)
Ginsburg 8.3% MeOH* Capillary
16ºC/min from 4
to -35ºC
Room
temperature
51 ± 36%**
BSMIS 10% DMA* Capillary
Placed on dry ice
for 30 min
Dilution in 20-x
volume extender
9-14%
Ginsburg 8.3% MeOH Cryovial
Placed on dry ice
for 20 min
33ºC for 8 s 28 ± 18%
HBSS* 8% MeOH
French
straw
10ºC /min from
5 to -80ºC
40ºC for 5 s 33 ± 20%
Medaka (Oryzias latipes)
FBS* 10% DMF* Cryovial
Nitrogen vapor
for 10 or 20 min
30ºC for 0.5-1
min, in 2-x BSS
96-100%
0.6 M
sucrose
10% DMSO* Capillary
Placed on dry ice
for 20 min
Holding between
fingers
85%
Green swordtail (Xiphophorus helleri)
HBSS
240-300
10% DMSO*
French
straw
45ºC /min from
5 to -80ºC
40ºC for 7 s
29 ± 8%***
HBSS300 14% glycerol
French
straw
20-35ºC/min
from 5 to -80ºC
40ºC for 7 s 77 ± 3%***
HBSS310
HBSS500
14% glycerol
French
straw
25ºC /min from
5 to -80ºC
40ºC for 5 s
1-3 of 15
females
Monterrey platyfish (Xiphophorus couchianus)
HBSS
240-300
14% glycerol
French
straw
25ºC /min from
5 to -80ºC
40ºC for 7 s 78±3%***
HBSS500 14% glycerol
French
straw
25ºC /min from
5 to -80ºC
40ºC for 5 s
2 of 15
females
* MeOH, methanol; DMA, dimethylacetamide; DMF, dimethylformamide; DMSO, dimethyl
sulfoxide; HBSS, Hanks’ balanced salt solution; FBS, fetal bovine serum.
** Relative to the control (71 ± 5%); actual rate was: 0.71*51 = 36 ± 26 %.
*** Post-thaw motility only, no fertilization data available.

STRATEGIES OF NILE TILAPIA (Oreochromis niloticus) POND CULTURE FOR SMALL-
SCALE FARMERS
Yang Yi and James Diana
College of Fisheries and Life Science
Shanghai Ocean University
Shanghai, China
yiyang@shou.edu.cn
Different strategies for Nile tilapia (Oreochromis niloticus) culture in ponds with a series of progressive inputs were compared.
The sequential experimental stages to increase fish production through intensification were: 1) triple superphosphate (TSP) only;
2) chicken manure only; 3) chicken manure supplemented with urea or urea and TSP; 4) urea and TSP; 5) continually supplemental
feeding; 6) staged supplemental feeding; 7) feeding alone.
The results showed that the choices of input regimes with increasing economic gains are: 1) fertilizing ponds with moderate loading
of chicken manure; 2) fertilizing ponds with chicken manure supplemented with urea and TSP; 3) fertilizing ponds with urea and
TSP; 4) fertilizing ponds initially with urea and TSP in combination of supplemental pelleted feed at 50% satiation level at later
stage of grow-out cycle.
SURVIVAL ANALYSIS METHODS FOR AQUACULTURE RESEARCH
Kathleen M. Yeater
USDA-ARS-Southern Plains Area
1001 Holleman Drive East
College Station, TX 77840 USA
kathleen.yeater@ars.usda.gov
Survival analysis is a class of methods for which the outcome variable of interest is time until an event occurs. Death or failure
is commonly considered the “event”, but any biological occurrence can be studied within this application. The flexibility of
these methods also allow for modeling multiple (or recurrent) events, such as change in disease stage.
Aquaculture research examples will be addressed on appropriate methods for estimation and interpretation of survival using
the Kaplan-Meier method, comparison of survival among different groups, relationship assessment of time-independent and
time-dependent explanatory variables, and capability of prediction of time until the event.
Oftentimes, conventional statistical methods such as logistic regression or linear regression are inappropriately used in analysis.
This presentation will also discuss selecting the appropriate analysis in terms of time-dependent variables, censored observations,
and non-normal distributions.
3 3

3 4
STUDING THE ULTRA STRUCURAL IN LIVER AND KIDNY BY USING ELECTRON
MICROSCOPY AS WELL AS HEMATOLOGICAL PICTURE AND MEASURING
BLOOD PARAMETER DUE TO EXPERIMENTAK INFECTION WITH FLEXIBACTER
COLUMNARIS IN MONOSEX TILABIA
Dr. Adel Younes
Department of Veterinary Services
Egypt
A total number 25 apparently health Monosex tilapia fish average body weigh 70 gm . five fish were randomly sampled and
submitted to bacterial isolation for verification of absence of tested bacteria .The other 20 fish divided into two group 10fish
for each group first 10 fish were injected intramuscularly with 1 ml of virulent strain of flexibacter columnaris adjusted at
McFarland No.2 (approximality 6x10
.
-8 cell /ml . The second group10 fish were injected with 1 ml sterile saline and served
as control. The site of injection was dorsal muscle just behind the head .Aeration was maintained and temperature ranged from
25 – 27 C according to Baxa et al (1987).The clinical signs and PM lesions were observed during the experiment .Specimens
from liver kidney spleen and gills were collected and use for histopathological studies .Moreover blood samples were collected
for determined hematological picture and blood parameters .Sample from liver kidney were taken at zero time from control
group and at 12.24,48,72and 96 hours for ultra structural three infected fish were used in case of each sample. The period of
this experiment was 96 hours
Fig. (1) Differential leucocytic Fig. (2) Serum protein constituent Fig. (3): Serum enzyme level
Fig. (4). Liver of Monosex tilapia 72 hours Fig (5). Kidney of Monosex tilapia 12 hours post
post injection-showing apoptosis(x 4000). injection-showing Mitochondrial swelling (M),
formation of myelin fibers (↑), loss of cytoplasmic
matrix and rough endoplasmic reticulm (RER)
, (x10000)

AN ORAL DELIVERY SYSTEM FOR DNA VACCINES IN RAINBOW TROUT
Yan Zheng, John Eley, Ram Veerubotla, Sophie St-Hilaire , Marie Adomako, Wendy Sealey, Peter Sheridan,
Malcolm Shields, Brian Donahower and Scott LaPatra
Department of Pharmaceutical Sciences
Idaho State University
970 South 5th Ave.
Pocatello, ID USA
zhenyan@isu.edu
A DNA vaccine for infectious hematopoietic necrosis virus has been licensed in Canada for use in Atlantic salmon. This
vaccine is effective against the virus, but is not being investigated for use in the U.S. rainbow trout industry because of the
high cost of administering the vaccine intramuscularly. We have evaluated the use of Poly (D, L-lactide co-glycolide) polymer
(PLGA) particles to deliver DNA plasmids to fish. Particles were created which incorporated approximately 40 % of the initial
DNA. Concentrations of DNA were estimated at more than 3 ug/mg of particles, and particle size was, on average, ~600 nm.
Preliminary results from our oral and anal intubation trials suggest particles are found within the lamina propria of the lower
intestine within 24 hours with both delivery methods. Further, particles top coated on commercial trout feed pellets were also
found in the lamina propria within 48 hours.
3

386
AQUACULTURE AMERICA 2009
ABSTRACT ADDENDUM
WEANING TIME AND THE FREQUENCY AND AMOUNT OF Artemia IN SPOTTED
SEATROUT (Cynoscion nebulosus) LARVICULTURE
Angelos Apeitos*, Jason T. Lemus and Reginald B. Blaylock
Thad Cochran Marine Aquaculture Center
The University of Southern Mississippi
Gulf Coast Research Laboratory
Ocean Springs, MS 39564 USA
angelos.apeitos@usm.edu
The spotted seatrout, Cynoscion nebulosus is a popular sport and food fish in the northern Gulf of Mexico. The Gulf Coast
Research Laboratory is seeking to improve seatrout larviculture techniques for both stock enhancement and potential
commercial applications. Weaning and cannibalism are the major challenges in seatrout larviculture. Weaning onto dry food
is a lengthy process during which cannibalism exacts a significant toll. Thus, successful seatrout larviculture requires a large
dependence on Artemia throughout the 25-day larval period. The standard protocol begins weaning by 8 days post-hatch (DPH)
and completes it between 22 and 23 DPH, during which Artemia are fed 6 times daily in excess of 14/mL/day. Two preliminary
studies were performed to assess the effect of 1) weaning larvae onto dry food on or before 11 DPH on survival, length and
condition at 18 DPH and 2) Artemia density and feeding frequency on survival through 24 DPH as well as notochord length,
condition (myotome height/notochord length) and survival following a salinity shock (stress test) on 24 DPH. Newly hatched
seatrout were stocked at 10/L in 27C 25ppt seawater and fed 5-15 rotifers/mL/day on 2-10 DPH.
The effect of weaning time was assessed in twenty 25-L circular tanks within a recirculating aquaculture system. Dry food and
Artemia ranging from 1-6/mL/day were fed beginning 6 DPH and Artemia were removed from the diet at either 7, 9, or 11 DPH
or not removed at all (control). The ration of Artemia was split between two feedings and dry food was fed up to five times
daily. Each feeding condition was replicated 5 times. Control fish survival and length were significantly greater than all other
groups, which did not differ from one another at 18 DPH. Condition was unaffected by weaning time. The results suggest that
the dry diet was nutritionally adequate, but the cannibalistic nature of seatrout larvae prevents early weaning.
The effects of Artemia density and feeding frequency were examined in a factorial study using thirty 60-L circular tanks
within a recirculating aquaculture system. Artemia were fed beginning 8 DPH at a maximum of 10, 14, or 18/mL either 2 or 6
times daily. Each combination of frequency and density was replicated 5 times. Dry food was provided 6 times daily on 12-24
DPH. There was no significant effect of frequency or density of Artemia fed on survival, stress tolerance, notochord length, or
condition of seatrout juveniles at 24 DPH. The results indicate that seatrout husbandry could be simplified by feeding Artemia
twice daily rather than six times. Future studies should focus on examining the feasibility of weaning sometime between 11 and
24 DPH and appropriately matching the density of Artemia to the age of seatrout larvae.

AQUATIC RESOURCE USE AND CONSERVATION FOR SUSTAINABLE FRESHWATER
AQUACULTURE AND FISHERIES IN MALI
James Bowman*, Héry Coulibaly, Charles Ngugi, Yang Yi, Nancy Gitonga, Lisa Reifke and Hillary Egna
AquaFish Collaborative Research Support Program
Oregon State University
Corvallis, OR 97331-1643 USA
james.bowman@oregonstate.edu
The AquaFish Collaborative Research Support Program is partnering with the Direction Nationale de la Pêche (Government of
Mali) and collaborators from Moi University (Eldoret, Kenya), Shanghai Ocean University (Shanghai, China), and FishAfrica
(Nairobi, Kenya) to implement an innovative project for the sustainable development of the aquaculture and fisheries sectors
in Mali. Support for this project is provided through a cooperative agreement between the Mali Mission of the United States
Agency for International Development (USAID) and Oregon State University.
The project takes a South-South approach, working in three theme areas with corresponding objectives as follows:
• Theme I (“Pond Culture”): Advancing Sustainable Freshwater Aquaculture Practices and Technologies; Objective:
To identify appropriate strategies for pond aquaculture and make them available to farmers in target areas
• Theme II (“Rice-Fish”): Promoting Sustainable Rice-Fish Aquaculture in Irrigated Systems; Objective: To identify
appropriate strategies for integrated rice-fish farming and make them available to farmers in target areas
• Theme III (“Fisheries Planning”): Building Community and Consensus towards a Fisheries Management Plan;
Objective: To help develop appropriate fisheries management plans to ensure long-term viability and sustainability of
capture fisheries in target areas
The project achieves its objectives largely through training, field trials and demonstrations, and stakeholder workshops. Theme
I provides hands-on training in pond construction, fish propagation, and pond management, and conducts field trials to identify
pond culture systems suitable for implementation in Mali. Theme II provides field testing and training to evaluate appropriate
adaptations of rice-fish systems for introduction into irrigated systems in Mali’s Niger River delta, and Theme III assists in
conducting frame surveys and involving local fishing groups in the development of sound fisheries management agreements,
working initially in the Lake Sélingué area. Most activities are carried out in Mali, but some training also occurs in China and
Kenya. The first short course, dealing with appropriate rice-fish culture techniques (Theme II), was held in China in September
of 2008, and the initial workshops for Themes I and III are scheduled to be held in Mali in January and February of 2009.
This poster provides background and justification for the project and outlines the current and planned activities being undertaken.
387

388
AQUACULTURE OF THREESPINE STICKLEBACK Gasterosteus aculeatus FOR USE IN
REASERCH
Mark C. Currey and William A. Cresko
Center for Ecology and Evolutionary Biology
335 Pacific Hall
5289 University of Oregon
Eugene, OR 97403-5289 USA
Aquatic model organisms have recently become very useful for studies of genetics, and zebrafish have been particularly
useful for studies of early vertebrate development. Research involving many other teleosts is beginning to blossom. One
of these teleosts is the threespine stickleback (Gasterosteus aculeatus). We use this fish at the University of Oregon to
elucidate the molecular basis of evolving traits. These traits include bony armor, craniofacial bone morphology and photoperiodic
response, among others. Although much of our work is performed on natural populations, we needed to establish laboratory
lines for developmental and genetic analysis.
To satisfy this need we designed and implemented an aquaculture technique to raise stickleback in the laboratory. These
approaches borrow from the vast knowledge of zebrafish aquaculture but are adapted to the unique requirements of stickleback.
In this poster I will compare the similarities and differences between zebrafish and stickleback approaches and systems,
present the requirements for successful husbandry of stickleback, and detail the stickleback-specific techniques that we have
developed.
SURVEY ON THE GROWTH RATE OF FARMED GREAT STURGEON FINGERLINGS
(Huso huso) IN FIBER GLASS TANKS BY USING ARTIFICIAL FEED
Gholamreza Darvishi, Mohammad Mazandarani, Narges Soleymani, Niaz Mohammad Kor and Lalik Sarikhani
Inland waters Aquatics Stocks Research Center
Gorgan
In this investigation the growth rate of farmed Huso huso intensively in fiberglass tanks by using of artificial feed was studied.
Fingerlings with average initial body weight of 8 grams prepared, then transported to 2m 2 tanks with 80 individuals per tanks.
The water prepared from well and the average water temperature was 19/28±4/6 and the average dissolved oxygen
concentration was 7/23±0/69 during the experimental period (8 months). Biological factors such as total length and weight
measured monthly. During the study, fingerlings with average weight of 8 grams and length 9/5 cm reached to average weight
of 293 grams and 43 cm respectively.
Weight gain, special growth rate, food conversion ratio and conditional factor were 285, 5%, 2/45 and 0/5% respectively during
this experiment.

DIFFRENTLY EXPRESSED ANTIOXIDANT GENES IN DISK ABALONE EXPOSED TO
HEAT STRESS
Mahanama De Zoysa* and Jehee Lee
Department of Marine Biotechnology
College of Ocean Science
Cheju National University
Jeju Special Self Governing Province
690-756, Republic of Korea
mahanama_dezoysa@yahoo.com
High temperatures represent one of the main physiological stresses, which foster elevated ROS levels and induced antioxidant
enzymes in marine organisms. Overall antioxidant defense in an organism is a combination of multiple responses of these
enzymes. Therefore, it is important to understand the functional relationship between different antioxidant enzymes against
heat stress.
This study was conducted to investigate responses of different antioxidant enzymes in mollusk disk abalone (Haliotis
discus discus) against oxidative stress upon exposure to high temperature above the habitat temperature. Disk abalones were
exposed to heat stress at 28 0 C for 48 h and control group was maintained at 20 0 C separately. Gill tissue was isolated in
different time points (1 h, 2 h, 4 h, 8 h, 12 h and 24 h) from both heat stress and control animals. Quantitative real time PCR was
conducted to analysis the heat stress responses of six anitioxidant enzymes including Mn-superoxide dismutase (Mn-SOD),
CuZn-superoxide dismutase (CuZn-SOD), catalase, thioredoxin peroxidase (TPx), mitochondrial thioredoxin-2 (MtTRx-2)
and Selenium dependant glutathione peroxidases (SeGPx).
Transcripts of the all six antioxidant enzymes were expressed in untreated abalone gill, mantle, abductor muscle, gonad,
digestive tract tissues and hemocytes with tissue specific manner. Time frame heat stress experiment results showed that
transcriptional responses were varied in different antioxidant enzymes. Transcriptional up-regulation of MnSOD (2.07-fold at
1 h), catalse (3.07-fold at 8 h), TPx (2.8-fold at 12 h) and SeGPx (13.2-fold at 24 h) were observed in abalone gill tissue after
heat stress at 28 0 C compared to control animal at 20 0 C. In contrast CuZnSOD and Mit-TRx-2 transcripts were initially down
regulated until 4 h and then reached to basal level at 24 h.
Our results revealed that MnSOD has immediate heat stress response at 1 h, while SeGPx has slow response at early stage
however it showed the highest expression (13.2-fold) at 24 h compared to other antioxidant enzymes in abalone. Finally, our
preliminary results suggest that abalone antioxidant enzymes can corporately response to the oxidative stress generated high
heat condition, however exact interrelations of these enzymes need to be clarified in future.
389

390
COMPARISON OF EXTRACTED ALGINIC ACID IN BROWN ALGAES; Sargassum
illicifolium, Cystoseira indica and Nizimuddinia zanardini IN THE OMAN SEA (CHABAHAR)
Abdollah Haghpanah, Lalik Sarikhani, Yousof Irii and Behrooz Gharavy
Fishery Research Center Of Golestan
This study was determined alginic acid in three species of brown seaweeds (Sargassum illicifolium , Cystoseira indica and
Nizimuddinia zanardinii) during autumn and winter 1384. Seaweed samples were colected from sea beach and washed in
fresh water and dried in the sun. So were under Formaldeid (0.5%) and Solforic acid (0.2 N) Respectively (2,5 hours). Fainaly
were under Alkaloid extraction by sodium carbonate (5%), that were obtined fluid after filteration. The Alginic acid resulted
with chloridric acid (1N). The extraction yield of alginic acid in autumn for Sargassum were 32.1±0.96 , Cystoseira 23.1±0.46
and Nizimuddinia 28.3±0.61 percent , and in winter for Sargassum were 29±0.72 , Cystoseira 21.4±0.53 and Nizimuddinia
24.3±0.32 percent.
According to this results, One way ANOVA showed that average of Alginic acid were not similar and significant differences
(P< 0.05 ) between species.
CULTURE OF Gracilaria corticata IN THE EARTHEN POND AND SEA (CHABAHAR)
Abdollah Haghpanah, Lalik Sarikhani, Yousof Irii and Behrooz Gharavy
Fishery Research Center of Golestan
The study of Gracilaria cultuer was carried out from January 2000 to June 2001 during 2 seasons in the earthen ponds and sea
water. Culture system was fixed bottom line method in pond and floating raft method in sea. In this study 500 gram gracilaria
were cultured on the line and investigated about 9 weeks. Gracilaria reached to maximum growth between 5 to 7 weeks and
biomas were 2.58 times in the winter, 2.64 times in spring in the pond and 2.7 in winter and 3.34 in sprin in sea water. One way
ANOVA showed that average weight was not similar in various season and showed significant difference. The avearage daily
growth rate in different season was between 4.3 to 35.7 grams too.

EFFECT OF SALINTY, AMMONIA AND FORMALINE ON IMMUNE, METABOLIC AND
HISTOPATHOLOGY MARKERS IN THE SHRIMP Litopennaeus schmitti
Raico Ernesto Laria Lamela, Yanis Cruz, Raquel Silveira, Mercedes Martinez and Norma Gonzalez
Several effectors of the immune response in invertebrates have been
considered as potential health or disease markers in crustaceans. The
relative simplicity of the invertebrate’s immune system makes it a
potential means of monitoring the effects of environmental contaminants
and the complex interactions that ultimately affect host resistance. Cellular
defences in crustaceans rely on haemocytes, with several functions such
as coagulation, phagocytosis, and encapsulation and wound healing.
The proPO activating system plays an important role in the recognition
and defence against invading foreign organisms and it is the enzyme
responsible for initiating the biosynthesis of melanin. Peroxidase activity
is associated with an adhesion molecule called peroxinectin which acts as
an opsonin, a degranulation and encapsulation-promoting factor as well
as having peroxidase and cell adhesion activity. It may function as an
efficient microbicidal attack system against invader microorganisms.
Salinity is one of the main environmental factors that wield a selective
pressure on aquatic organism. Formalin is used in shrimp industry as
an external parasiticide to control protozoan parasites. Ammonia is the
major end product of protein catabolism excreted by fish, crustaceans,
and molluscs and the accumulation of ammonia can cause mortality of
organisms reared in closed culture systems
The main objectives of this work were to determine phenoloxidase and
peroxidase activity, total haemocytes count and histopathology changes
and the behaviour of metabolic indicators in the haemolymph of the
white shrimp Litopennaeus Schmitti exposed to low salinity, formalin and
different levels of ammonia.
391

392
CULTURE OF Gracilaria corticata IN THE EARTHEN PONDS OF BERIS SESSION
(CHABAHAR)
Abdollah Haghpanah, Lalik Sarikhani, Yousof Irii and Behrooz Gharavy
Fishery Research Center of Golestan
The study of Gracilaria cultuer was carried out from October 2000 to Sebtember 2001 during 4 seasons in the earthen ponds
. Water depth in ponds was 110 Cm. Culture system was fixed bottom line method. In this study 500 gram gracilaria were
cultured on the line and investigated about 9 weeks. Gracilaria reached to maximum growth in 6 to 7 weeks and biomas were
2.3 time in the atumn, 2.6 time in winter, 2,7 in spring and 2.4 in summer respectivaly. One way Anova showed that average
weight was not similar in various season and showed significant difference. The avearage daily growth rate in different season
was between 4.3 to 34.3 grams too.
CLONING, EXPRESSION AND PURIFICATION OF BETA-AGARASE II FROM
Saccharophagus sp. AG21
Youngdeuk Lee, Kwangsun Jung, Jihee Lim, Se Jae Kim and Jehee Lee
Department of Marine Biotechnology
College of Ocean Science
Cheju National University
Jeju, 690-756, Republic of Korea
Agar is an important gelifying agent for biochemical use and the food industry. Agar is a polysaccharide which linked
D-galactose and L-3,6-anhydro-galactose residues. Beta-agarase cleave 1,4-β-linkage. The named beta-agarase β -agarases are
classified into two groups named as β -agarase I and II based on their cleavage of β-(1→4) linkages. Main hydrolyzed products
of β-agarase I are neoagarotetrose and neoagarohexaose, while β -agarase II produces the neoagarobiose.
We screened agarase producing marine bacteria from seawater of Jeju island. We isolated genomic DNA from the bacteria
and analyzed 16s rRNA sequence by PCR and sequencing. The sequence was 99% (1376/1384 bp) similar to Saccharophagus
degradans 2-40. We determine the ORF sequence of agarase (agaY2). We Clone, over expression and purification of
recombinant protein using E. coli expression system. We analyze reaction products of recombinant protein by thin layer
chromatography.
The 2334 base pair ORF of agaY2 agarase is encoding 778 amino acid residues. The predicted agaY2 has molecular mass of
88 kDa and and isoelectric porint 5.2. Pair wise amino acid comparison of agarase showed 96% highes level of identity with
Saccharophagus degradans 2-40 agarase. The purified recombinant agaY2 was not able to degrade agar, however it could
only degrade the neoagarotetraose and neoagarohexaose, mainly into neoagarobiose components. AgaY2 showed functional
character of beta-agarse II.

GROTH PERFORMANCE AND RESISTANCE TO Strptococcus iniae OF NILE TILAPIA
Oreochromis niloticus FED VARIOUS DIETARY LEVELS OF THIAMIN
Chhorn Lim * , Mediha Yildirim-Aksoy and Phillip Klesius
Aquatic Animal Health Research Laboratory
USDA-ARS, MSA
Auburn, AL USA
Chhorn.lim@ars.usda.gov
Thiamin or vitamin B 1 functions in all cells as the coenzyme thiamin pyrophosphate that involves in the oxidative decarboxylation
of α-keto acids and the transketolase reactions in the pentose phosphate pathway. Thiamin has been demonstrated to be
essential in diets of fish, and quantitative requirements have been determined for several fish species including rainbow trout,
Pacific salmon, common carp, channel catfish, turbot, yellowtail and tilapia. Results of the few studies with homeotherms on
the effect of thiamin on immune function and disease resistance are contradictory. However, to our knowledge, no studies have
been conducted on the effect of dietary levels of thiamin on disease resistance in fish. Therefore, this study was conducted
to evaluate the effect of dietary levels of thiamine on growth performance, immune response and resistance of Nile tilapia
(Oreochromis niloticus) to Streptococcus iniae challenge.
An 8-week feeding study was conducted in flow-through 57-L aquaria with juvenile Nile tilapia (3.6 ± 0.2 g) stocked at a rate
of 35 fish/aquarium. Casein (vitamin-free)-gelatin based diets supplemented with 0, 2, 4, 8, 16 and 32 mg thiamin/kg were each
fed to fish in four replicate aquaria twice daily to apparent satiation. All fish in each aquarium were counted and weighed at 4
week intervals. At the end of 4, 6 and 8 weeks, 2, 2 and 3 fish/aquarium, respectively, were sampled and bled for determination
of serum pyruvate and lactate. At the end of week 8, another 3 and 4 fish/aquarium were bled for hematological and serum
immunological assays, respectively. Eighteen fish/aquarium (17 only for fish fed the diet without supplemental thiamin) were
intraperitoneally injected (IP) challenged with 100 µL of 1 x 10 4 CFU/mL of S. iniae and mortality was recorded daily for
14 days. At the end of the challenge period, four surviving fish/aquarium were bled for measurement of serum agglutinating
antibody titers against S. iniae.
Results showed that Nile tilapia fed the thiamin-unsupplemented diet had significantly (P < 0.05) reduced weight gain, feed
intake, feed efficiency and survival relative to fish in other treatments. These parameters did not differ among fish fed diets
supplemented with 2 mg thiamin/kg or more. Body protein did not differ among treatments, but lower body lipid and higher
moisture and ash were obtained in fish fed the diet without thiamin supplementation. Fish fed this diet also had significantly
lower hematocrit than those fed thiamin supplemented diets, but other hematological variables were not influenced by dietary
levels of thiamin. Serum pyruvate at weeks 4, 6 and 8 significantly increased in fish fed the thiamin-unsupplemented diet but
did not differ for fish fed other diets. Serum lactate, however, was not affected by dietary levels of thiamin. Serum protein, total
immunoglobulin, lysozyme and alternative complement activity, and post-challenge antibody titer were not affected by dietary
treatments. Cumulative mortality 14 days post-challenge was significantly higher in fish fed the thiamin-unsupplemented diet
but did not differ among fish in other treatments.
393

394
CHARACTERIZATION OF THERMOSTABLE BETA AGARASE AG17 FROM MARINE
BACTERIA Agarivorans sp. AG17
Chamilani Nikapitiya*, Chulhong Oh, Youngdeuk Lee, Ilson Whang and Jehee Lee
Department of Biotechnology
Cheju National University
66, Jeju Self Governing Province
690-756, Republic of Korea
chamilaninikapitiya@yahoo.com
Agar degrading bacteria Agarivorans sp. AG17 was isolated from red seaweed Grateloupia filicina at Jeju coast, Rep. of Korea.
Gene for beta agarase was cloned, sequenced and characterized from identified bacteria strain and named as AG17. It consists
of 2985 bp open reading frame encoding 995 amino acids. The predicted molecular mass of the mature protein was 105 kDa
and Isoelectric point (Ip) was 4.1. The amino acid sequence of AG17 belongs to the glycoside hydrolase family (GHF) 50. The
C-terminal transmembrane helices sequence was identified at the amino acid position from G 968 -L 990 . ClustalW pairwise identity
results revealed that AG17 has the highest nucleotide (98.6%) and amino acid (99.7%) identity to beta-agarase sequence of
Agarivorans sp. JAMB-A11 (agaA11).
Recombinant AG17 was over expressed using E. coli bacterial expression system and protein was purified. The purified
enzyme was functionally active and showed higher thermal and pH stabilities. The enzyme could produce neoagaro-
oligosaccharides such as neoagarohexaose and neoagarotetraose by degrading agar and finally could degrade those into
neoagarobiose suggesting that enzyme was endo beta agarase type. Therefore, it could be suggests that purified AG17 enzyme
would have potential for the industrial applications such as development of cosmetics and pharmaceutical industry.

EFFECTS OF LOADING DENSITY ON GOLDEN SHINER SURVIVAL
Philip Pearson*, Brian Small, Rachel Beecham, Todd Sink, Susan LaBarre and Doug Minchew
USDA-Agricultural Research Service
H.K. Dupree Stuttgart National Aquaculture Research Center
P.O. Box 1050
Stuttgart, Arkansas 72160 USA
philip.pearson@ars.usda.gov
We conducted four hauling trips of 6 h each to investigate effects of loading density (120, 180, and 240 g fish/L) on survival of
golden shiners Notemigonus crysoleucas. Commercially graded golden shiners (mean weight 3.3 g ± 0.04 SE) were transported
in insulated hauling tanks filled with fresh well water, chilled with unchlorinated block ice, and aerated with pure oxygen.
Whole-body cortisol analyses were not statistically different for golden shiners transported at 120, 180, or 240 g fish/L. Unionized
ammonia concentration increased with respect to loading density, but had no apparent effect on survival. Results of this
study indicate golden shiners can be successfully (≥ 99 % survival) transported for up to 6 h at 240 g fish/L in well water chilled
with unchlorinated block ice and aerated with pure oxygen.
Table 1. Mean (± SE) hauling survival, survival of 1 kg of fish transferred to tanks and held for 18 h after
hauling, whole body cortisol, total ammonia nitrogen (TAN), and un-ionized ammonia (UIA) of golden
shiners before and after hauling at three loading densities. Different letters within a column indicate
treatment means that were significantly different (P ≤ 0.0001)
Treatment N Survival Survival Cortisol TAN
UIA
(end of
trip)
(18 h post delivery)
(ng/g)
Before Hauling 4 N/A N/A 15.7 ±
120 g of fish/L 8 99.5 ± 0.1 99.9 ± 0.1 15.0 ±
180 g of fish/L
7*
1.6
1.3
99.5 ± 0.1 99.9 ± 0.02 17.5 ±
240 g of fish/L 8 99.4 ± 0.1 99.9 ± 0.02 19.9 ±
2.3
2.1
(mg/L)
(mg/L)
1.68 ± 0.20 z 0.06 ± 0.01 z
5.64 ± 0.30 y 0.10 ± 0.02 y
8.93 ± 0.20 x 0.13 ± 0.01 x
10.66 ± 0.04
w
0.20 ± 0.04
ANOVA P = 0.51 P = 0.46 P = 0.27 P < 0.0001 P < 0.0001
*On day 1 (May 29), one replicate of the 180 g/L treatment had 32% mortality due to a low dissolved
oxygen concentration (0.4 mg/L) attributed to equipment failure. This replicate was not included in the
statistical analysis.
w
395

396
MANGROVES LOSS-A GLOBAL PERSPECTIVE
Alfredo Quarto
Executive Director
Mangrove Action Project
www.mangroveaction project.org
Mangroves are the rainforests by the sea. These unique coastal wetlands are essential habitat and nurseries fo a wide variety
of sea life, including fish, crudtaceans and molluscs. It is believed that two thirds of tropical and sub-tropical marine species
spend some part of their lives in the mangrove wetlands, which also serve as important habitat for migratory birds and sea
turtles. As well, mangroves filter out upland pollutants via their intricate root systems, and protect coral reefs and sea grass beds
from sedimentation. Large stretches of the sub-tropical and tropical coastlines of Asia, Africa, Oceania, the Americas and the
Caribbean are fringed by mangroves, once estimated to cover an area of over 32 million hectares. Now, less than 15 million ha
remain-less than half the original area.
The importance of the protective mangrove buffer zone cannot be overstated. In regions where these coastal fringe forests have
been cleared, tremendous problems of erosion and siltation have arisen, and terrible losses to human life and property have
occurred due to destructive hurricanes, storm surges and tsunamis.
Today there is a growing urgency to recognize the importance of conserving and restoring protective mangrove greenbelts to
lessen the dangers from future catastrophes, because as sea levels rise so will the frequency and intensity of hurricanes and
storm surges. Mangroves can buffer against the fury of such destructive storms, protecting those settlements located behind a
healthy mangrove fringe.
Mangrove Action Project (MAP) is working with other organizations in the global South towards restoring degraded and
cleared mangrove areas as a high priority. MAP is especially interested in restoring some of the 250,000 ha of abandoned
shrimp farms located in former coastal wetland areas, especially in Asia and Latin America.

STUDIES WITH HEAT SHOCK PROTEIN STIMULATION FACTOR IN STRESS
MANAGEMENT IN FIN FISH CULTURE
Ronald J. Roberts*, Carmelo Agius and Charles Saliba.
Hagerman Aquaculture Research Laboratory
Hagerman, Idaho 83332 USA
heronpisces@btinternet.com
Heat Shock Proteins, otherwise known as cellular chaperones are guardian proteins, highly conserved throughout evolution and
found intra and extra-cellularly in all living creatures. They are produced in response to all stressful conditions (not just heat)
and help individual cells cope by supporting the cellular processes such as protein production and cellular membrane integrity
and also facilitate apoptosis and cellular recycling. Their activities can be broadly expressed as inhibiting undesirable actions
and interactions within the cell and promoting the desirable ones. In this respect they also play a role in immunogenesis and in
control of neoplasia. There are four major families of HSP designated according to molecular weight (kDa) as Hsp90 Hsp70
Hsp 60 and Hsp30.
In humans they have been shown to be highly active in numerous stressful situations eg. deep-sea divers, distance athletes and
following surgery. Until recently however, it has not been possible to induce them in advance of a stress and so help to obviate
the stress ab initio.
Recently a patented plant extract, Tex-OE has been shown to contain a specific Heat Shock Protein Stimulation Factor. As the
commercial product “Pro-Tex” this has been shown to mediate stress in humans, poultry, commercial rabbits and race-horses
and is commercially available in Europe for human and animal use as a neutraceutical. In UK, poultry chicks are now routinely
exposed to Pro-Tex in the first week of life, to ensure stress reduced hatchery output.
Studies with Pro-Tex as a component of feed and as an additive to the water, whence it is imbibed, have been carried out in
our laboratories and in commercial tilapia, salmon, trout, aquarium fish and sea-bass farms in USA UK and Malta. These have
included toxicity protection studies, transport stress studies, vaccination stress studies and studies on the effect of HSP’s on
sperm survival.
In all cases it has been shown that pre- exposure of fin fish to Heat Shock Protein Stimulation factor in the water or feed induces
high and persisting tissue levels of Heat Shock Proteins for up to three days, without any detrimental effects, and with often
remarkable effects if such fish are subjected to severe stress. In addition use prior to intraperitoneal vaccination reduces toxicity
and adhesions and samples of milt exposed to extender containing Pro-Tex is active and fertile for 50% longer than the same
milt in standard extender.
397