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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
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February 15-18, 2009 Washington Sta
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TABLE OF CONTENTS Preface..........
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STUDY On MORTALITY EGG Of Acipenser
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HEAVY METALS IN THE BIVALVE MOLLUSC
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THE POTENTIAL OF “LOST CROPS” I
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DIETARY CARBOHYDRATE LEVEL AFFECTS
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EFFICIENT AND RELIABLE PROTOCOLS FO
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MULITIVARIATE ANALYSIS OF THE GROWT
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NEW CONCEPTS FOR STUDY ON SURVIVAL,
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FORMULATION AND APPLICATON OF PROBI
Page 21 and 22:
ALTERNATIVE USES FOR RETIRED CATFIS
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EFFECT OF FEED RATE ON FOOD CONSUMP
Page 25 and 26:
UTILIZATION OF FISH PROCESSING BYPR
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STUDY OF LIMNOLOGICAL VARIABLES IN
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GROW-OUT PERFORMANCE OF NILE TILAPI
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INNOVATIVE USES FOR UNDER-UTILIZED
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STATISTICAL DESIGN AND ANALYSIS OF
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SEEPAGE LOSSES FROM EARTHEN AQUACUL
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EVALUATION OF BODY AND FILLET COMPO
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EFFECTS OF PHEROMONAL STEROIDS OR P
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EFFECTS OF DIETARY PREBIOTICS ON MI
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SOLDIER FLY PREPUPAE AS AN ALTERNAT
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ECONOMICS OF A SOFT-SHELL CRAYFISH
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ADVANCES IN THE CULTURE OF THE LANE
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DEVELOPMENT OF INTENSIVE CULTURE ME
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RECIRCULATING AQUACULTURE SYSTEM FO
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EVALUATION OF LARVAL FLORIDA POMPAN
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DEVELOPMENT OF A SELECTION INDEX ON
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EXPERIMENTAL USE OF ISOMETAMIDIUM C
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CHARACTERIZATION OF 14kDa APOLIPOPR
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EFFECT OF FEED DEPRIVATION AND INSU
Page 63 and 64:
WRITING “SMART” OBJECTIVES FOR
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AQUACULTURE ONLINE: HOW PRODUCTION
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ENERGY USE, RESOURCE CONSUMPTION, A
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COMPARISON OF ENERGY AND RESOURCE C
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DEVELOPMENT OF STANDARDS FOR FISHER
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THE EFFECTS OF STOCKING DENSITY AND
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EVALUATION OF DIFFERENT ORGANIC FER
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RECLAIMED AQUACULTURE EFFLUENT FOR
Page 79 and 80:
THE SYNERGISTIC USE OF DIETARY BIOT
Page 81 and 82:
SELLING AQUACULTURE PRODUCTS IN KEN
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WATER QUALITY AND TREATMENT EFFICIE
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ALTERNATIVE FEEDING STRATEGIES TO I
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EVALUATION OF SOLVENT EXTRACTED, DE
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WILL DECREASE IN POND BANK PRICES I
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OUTREACH, ACCEPTANCE, AND SUCCESS O
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EVALUATION OF A POINT OF CARE BLOOD
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REPLICATION IN FIELD AND LAB EXPERI
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TWICE-ANNUALLY SPAWNING RAINBOW TRO
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CAN CROWDING DENSITY AFFECT ON BODY
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CHANGES OF RELATIVE EGG WEIGHT DURI
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CONSUMER SAFETY ASSESSMENTS FOR CUL
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SEAFOOD AT ITS BEST: A FOUR-LESSON
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OPTIMIZATION OF SHRIMP DIETS USING
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EVALUATING POND AQUACULTURE EFFLUEN
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EVALUATION OF STRESS-INDUCED CORTIS
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PERFORMANCE OF Litopenaeus vannamei
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THE NEAH BAY GEODUCK AQUACULTURE AN
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POST-KATRINA MARINE BAIT DEVELOPMEN
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PHARMACOKINETICS OF FLORFENICOL IN
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DELTA 6 DESATURASE GENE IS DIFFEREN
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INFLUENCE INDIVIDUAL AND MIXED HEAV
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FORMALIN TREATMENT FOR SAPROLEGNIA
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APPLICATION OF SCALE COVER GENE N F
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THE EFFECTS OF HIGH VS. LOW DISSOLV
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SEA TRIALS OF A SELF-PROPELLED OFFS
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DEVELOPMENT OF A VALUE-ADDED PRODUC
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THE EMERGENCE OF VIRAL HEMORRHAGIC
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COMPREHENSIVE QUALITY ASSESSMENT OF
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LIVE BAIT SHRIMP MARKETING IN THE G
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SPERM MOTILITY ACTIVATION IN THE ES
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ENERGY EFFICIENCY AND ENERGY USE RE
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CONCENTRATION OF ORGANIC MATERIAL I
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VHS BIOSECURITY WORKSHOPS AND THE D
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PROSPECTS FOR ENGINEERING SOYBEANS
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ROE AND CAVIAR BACTERIOLOGY: In viv
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A ZEBRAFISH MODEL TO INVESTIGATE TH
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SIZE AT SEXUAL MATURITY AND FECUNDI
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EFFECT OF HERBAL LEAF DECOCTION ON
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AQAUFISH CRSP: FOSTERING THE DEVELO
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STUDY ON POSSIBILITY OF PRODUCTION
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HISTORY AND STATUS OF PADDLEFISH AQ
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AN EVALUATION OF CHANNEL CATFISH EG
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Macrobrachium rosenbergii NODAVIRUS
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EFFECT OF TOTAL REPLACEMENT OF FISH
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ON CHOICE AND USE OF STATISTICAL TO
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TISSUE DISTRIBUTION OF LEPTIN-LIKE
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PALM OIL EFFICACY IN OXIDATION STAB
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A SPREADSHEET MODEL FOR FINANCIAL P
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SUSCEPTIBILITY OF PACIFIC SALMONIDS
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ALTERNATIVE USES FOR ALGAE PRODUCED
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DEVELOPMENT OF SPF STOCKS OF BAIT S
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GROWTH PERFORMANCE OF ATLANTIC COD
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IDENTIFICATION, CLONING AND EXPRESS
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INTENSIVE CULTIVATION OF Acartia to
Page 191 and 192:
ANALYZING RESULTS THROUGH TIME USIN
Page 193 and 194:
SLOPE RATIO ANALYSIS IS A ROBUST AN
Page 195 and 196:
THE USE OF HYDROGEN PEROXIDE (H 2 O
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DETECTION OF YELLOW HEAD DISEASE IN
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EFFECTS OF STANDARD AND HIGH-FAT DI
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EFFECTS OF STANDARD AND HIGH-FAT DI
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OPTIMIZING FEEDING STRATEGIES FOR T
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ISOLATION, IDENTIFICATION AND BIOLO
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MODELLING OF NUTRITIONAL REQUIREMEN
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FATTY ACID COMPOSITION OF Artemia u
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VERIFICATION OF THE POTENCIAL AQUIC
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AQUAPLANT: A WEB-BASED TOOL FOR AQU
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EFFICIENT OXYGENATION FOR SUSTAINAB
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PRODUCTION OF Artemia CYSTS AND FLA
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VARIATION OF EGG SIZE OF DOMESTIC W
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BAITFISH AQUACULTURE IN THE MID-ATL
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Aeromonas hydrophila SEPTICEMIA IN
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PRELIMINARY NUTRITIONAL INVESTIGATI
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HYDROGEN PEROXIDE TREATMENTS FOR CH
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NUCLEOTIDE SEQUENCE VARIATIONS OF T
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RESPONSE OF PACIFIC WHITE SHRIMP, L
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SHRIMP RESEARCH ACTIVITIES AT OCEAN
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THE IMPACT OF CATFISH IMPORTS ON TH
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EFFECTS OF TWO DIETARY LIPID LEVELS
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NOVEL COMPOUNDS AND OPPORTUNITIES F
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HOLISTIC GOODNESS-OF-FIT: COMPARING
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DESIGNING A SUPPLY CHAIN FOR MARINE
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ONTOGENY AND CHARACTERIZATION OF SO
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AMBIENT CONDITIONING AND ULTRASOUND
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AQUACULTURE SAFETY IN KENTUCKY Tiff
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SUCCESSFUL CULTURE OF PINFISH Lagod
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PRELIMINARY STUDY OF THE EFFECTS OF
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METHYLMERCURY CONCENTRATIONS FOUND
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COMPARATIVE OXYGEN TOLERANCE OF BLU
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A STANDARD GENETIC STOCK OF RAINBOW
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ADVANCES IN THE DEVELOPMENT OF THE
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ORGANIC SHRIMP PRODUCTION: AN ATTEM
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EFFECT OF STOCKING DENSITY AND FEED
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EVALUATION OF THE EFFECTS OF TEMPER
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EVALUATE THE OF PLANKTONIC AND BENT
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RISK EVALUATION TO AQUACULTURE POND
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EVALUATION OF CHANNEL CATFISH (Icta
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GREENHOUSE GAS EMISSIONS ASSOCIATED
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MULTIPLE COMPARISONS OF MEANS VS. R
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PUTTING TOGETHER A BUSINESS PLAN St
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MORPHOLOGICAL DESCRIPTION OF THE IN
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DETERMINING SIZE EFFECTS ON PHOSPHO
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EVALUATION OF MULTIPLE-BATCH CHANNE
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ENGINEERING OPTIMAL SEED PHOSPHORUS
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EFFECTS OF PREBIOTICS ON NUTRIENT D
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EVALUATION OF SYNTHETIC ASTAXANTHIN
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EFFECT OF ZOOPLANKTON DENSITY ON GR
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CORN DISTILLERS DRIED GRAINS WITH S
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EFFECT OF TEMPERATURE AND SALINITY
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NOAA-USDA JOINT INITIATIVE ON ALTER
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EFFECTS OF TEMPERATURE REGIME ON NI
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ARACHIDONIC ACID (20:4n-6) EFFECT O
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GROWTH RATE OF JUVENILE AMAZONIAN P
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EVALUATION OF THE PREBIOTIC GROBIOT
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GENETIC DIVERSITY OF CULTURED AND W
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PROGRAM EVALUATION Michael H. Schwa
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SENSORY ANALYSIS OF RAINBOW TROUT O
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POULTRY BY-PRODUCT MEAL- BASED PELL
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SOME COMMON DESIGN AND STATISTICAL
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PADDLEFISH SYMPOSIUM DEDICATION TO
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FORECASTING POND BANK PRICES OF CAT
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C-TERMINAL MODIFICATION MEDIATES BI
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THE UTILIZATION OF HISTOPATHOLOGY I
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USE OF FRESHWATER LOW VALUE FISH FO
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GROWTH OF PACIFIC WHITE SHRIMP Lito
Page 331 and 332: DEVELOPMENT OF SMALL-SCALE AQUACULT
Page 333 and 334: The acuTe ToxiciTy of copper To lar
Page 335 and 336: GROWTH OF FED GOLDEN SHINERS IN AQU
Page 337 and 338: THE SAFETY OF COPPER SULFATE TO CHA
Page 339 and 340: PRACTICAL APPLICATION OF AN INTEGRA
Page 341 and 342: DEVELOPMENT OF PARATRANSGENIC Artem
Page 343 and 344: EFFECTS OF LOW-PHOSPHORUS DIETS ON
Page 345 and 346: TREATMENT OF RECIRCULATING WATER US
Page 347 and 348: A LOW COST SMALL SCALE PRODUCTION S
Page 349 and 350: DETERMINING MORPHOLOGICAL AND BIOCH
Page 351 and 352: DIETARY MAGNESIUM AND GROWTH OF PAC
Page 353 and 354: EVALUATION OF WATER QUALITY FOR LIV
Page 355 and 356: THE EFFECTS OF SOYBEAN OIL, FLAXSEE
Page 357 and 358: REPLACEMENT OF FISH MEAL WITH SOYBE
Page 359 and 360: SATURATED LIPID KEY TO MAXIMIZING F
Page 361 and 362: CHANGES IN WHOLE-BODY FATTY ACID CO
Page 363 and 364: EYE COLOR AS A PREDICTOR OF SOCIAL
Page 365 and 366: INCREASING LEVELS OF DIETARY CARBOH
Page 367 and 368: DEVELOPING VALUE-ADDED PRODUCTS FRO
Page 369 and 370: PROGRESS AND CONSTRAINTS TOWARD COM
Page 371 and 372: EFFECT OF CULTURE DENSITY ON PRODUC
Page 373 and 374: BAYESIAN METHODS FOR DETERMINING TH
Page 375 and 376: GROWTH PARAMETERS OF WILD AND SELEC
Page 377 and 378: QUALITY ASSESSMENT OF ATLANTIC STUR
Page 379 and 380: SMALL-SCALE, LOW-INPUT AQUACULTURE:
Page 381: IMMUNIZATION OF CATFISH WITH INACTI
Page 385 and 386: AN ORAL DELIVERY SYSTEM FOR DNA VAC
Page 387 and 388: AQUATIC RESOURCE USE AND CONSERVATI
Page 389 and 390: DIFFRENTLY EXPRESSED ANTIOXIDANT GE
Page 391 and 392: EFFECT OF SALINTY, AMMONIA AND FORM
Page 393 and 394: GROTH PERFORMANCE AND RESISTANCE TO
Page 395 and 396: EFFECTS OF LOADING DENSITY ON GOLDE
Page 397: STUDIES WITH HEAT SHOCK PROTEIN STI
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