Processed feed pea and canola meal for blue shrimp diets

Ž .
Aquaculture 196 2001 87–104
www.elsevier.nlrlocateraqua-online
Assessment of differently processed feed pea
ž Pisum satiÕum/ meals and canola meal žBrassica
sp. / in diets for blue shrimp žLitopenaeus
stylirostris/
L. Elizabeth Cruz-Suarez a , Denis Ricque-Marie a,) ,
Mireya Tapia-Salazar a , Ian M. McCallum b , David Hickling c
a Programa Maricultura, Facultad de Ciencias Biologicas, ´ UniÕersidad Autonoma ´ de NueÕo Leon, ´
Cd. UniÕersitaria Apdo. Postal F-56, San Nicolas de los Garza, NueÕo Leon ´ 66450, Monterrey, Mexico
b Pulse Canada, 330-360 Main Street, Winnipeg, Manitoba, Canada R3C 3Z3
c Canadian International Grains Institute. 1000-303 Main St., Winnipeg, Manitoba, Canada R3C 3G7
Received 8 June 2000; received in revised form 3 November 2000; accepted 6 November 2000
Abstract
The nutritional value of whole, dehulled, extruded, dehulled–extruded and micronized feed
peas, and extruded canola meal, included at a 30% concentration as ingredients in diets for
juvenile blue shrimp was assessed. Pea meals replaced a portion of soybean meal and wheat Ž1:3
parts.
on an isonitrogenous and isoenergetic basis, and similarly extruded canola meal replaced a
portion of soybean meal, fish meal and wheat Ž 1:2:3 parts.
of a control diet. After 28 days, the
shrimp almost tripled their weight on the experimental diets. Performance was unaffected by
dehulling, except for a slight increase in feed consumption and lower protein efficiency ratio
Ž PER .. Extrusion cooking had no effect on growth and survival but significantly improved feed
conversion and PER. The micronized pea diet produced the highest feed intake and growth rate.
Response to the diet containing extruded canola meal was similar to that of the control diet.
Ingredient apparent dry matter digestibility Ž IADMD.
ranged from 80.7% for micronized peas to
92.4% for dehulled–extruded peas, and was 79.4% for extruded canola meal. Higher IADMD
coefficients were obtained for extruded pea meals as a result of starch gelatinization. Ingredient
apparent protein digestibility Ž IAPD.
ranged from 79.1% to 85.4% and did not differ significantly
amongst the test meals. The test ingredients also conferred differential water stability properties
with the whole-extruded peas ingredient containing diet having the lowest dry matter and crude
) Corresponding author. Tel.: q52-8-3526-380; fax: q52-8-3526-380.
Ž .
E-mail address: lucruz@ccr.dsi.uanl.mx D. Ricque-Marie .
0044-8486r01r$ - see front matter q 2001 Elsevier Science B.V. All rights reserved.
Ž .
PII: S0044-8486 00 00572-X

88
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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104
Ž .
protein CP loss following a 1-h immersion in water. This study showed that whole raw feed pea
is a very acceptable ingredient for blue shrimp diets; extrusion cooking improved feed conversion
ratio and PER, and micronizing peas enhanced feed intake, while dehulling had no effect. q 2001
Elsevier Science B.V. All rights reserved.
Keywords: Crustacean; Nutrition; Growth; Digestibility; Peas; Canola
1. Introduction
Recent reports have described grain legumes or pulses, such as field peas as potential
ingredients for aquaculture feeds ŽAllan, 1998; Novoa and Castillo, 1998; Allan et al.,
1999a; Booth et al., 1999 .. Some reports have indicated that pea meal is an acceptable
ingredient in diets for rainbow trout ŽKaushik et al., 1993; Gomes et al., 1993, 1995;
Burel et al., 1996 ., tilapia Ž Fontainhes-Fernandes et al., 1997 ., silver perch ŽAllan, 1997;
Allan et al., 1999b; Booth et al., 1999 ., tiger shrimp Ž Smith et al., 1999.
and European
sea bass Ž Santos and Gomes, 1997; Gouveia and Davies, 1998, 2000 .. Peas offer
flexibility in feedstuff selection to the feed manufacturer as they may replace both grain
Ž e.g., maize corn, wheat. and protein Ž e.g., soybean meal and fish meal.
but their use has
been limited due to concerns about the presence of anti-nutritional factors ŽTacon,
1997 .. Studies with rainbow trout have shown that prior heat treatment was necessary to
improve digestibility and possibly inactivate anti-nutritional factors ŽKaushik et al.,
1993; Pfeffer et al., 1995 .. Feed peas, the designation for the round shaped, low tannin
varieties of Pisum satiÕum, are a major pulse crop marketed extensively in Europe and
Canada as a source of carbohydrate, mainly starch, and protein for livestock feeds
Ž UNIP-ITCF, 1995 .. As a result of plant breeding, several anti-nutritional factors in feed
peas including tannins and anti-trypsins, have been eliminated or substantially reduced
Ž Castell et al., 1996 ..
Starches are low cost sources of dietary energy and are generally well utilized by
shrimp Ž Catacutan, 1991; Shiau and Peng, 1992; Cruz-Suarez ´ et al., 1994 .. Thermal
processing to gelatinize starch increased its digestibility in diets fed to Litopenaeus
Õannamei Ž Davis and Arnold, 1993; Cousin et al., 1996 .. However, extrusion conditions
Ž none, wet or dry.
differentially affected energy digestibility coefficients of cereal grains
Ž Davis and Arnold, 1995 .. Dehulled peas had higher apparent digestibility coefficients
for energy, crude protein Ž CP.
and essential amino acids than whole peas when included
in formulated diets fed to silver perch Allan Ž 1997 .. Dehulling has also been shown to
improve the nutritional value of leguminous seeds as ingredients in diets for Penaeus
monodon Ž Eusebio, 1991 ..
Canola meal is a particularly rich plant source of sulphur containing amino acids for
fish diets Ž Higgs et al., 1995.
but its potential value as an ingredient in shrimp feed has
received little attention Ž Buchanan et al., 1997 .. Extrusion processing has also been
shown to improve the nutritive value of canola meal Ž Satoh et al., 1998 ..
The objective of the present study was to assess growth, nutrient utilization, survival
and digestibility in blue shrimp fed whole, dehulled, extrusion cooked and micronized
feed peas, and extruded canola meal, and held under laboratory conditions.

2. Material and methods
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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104 89
2.1. Diets
A lot of commercial dry peas P. satiÕum composed of mixed Canadian prairie
varieties was processed to prepare the respective meals. Whole and dehulled peas were
pin-milled to produce raw flours, WRA and DRA, respectively. Portions of these flours
were preconditioned Ž Wenger model 2 DDC.
and extruded with a co-rotating twin screw
extruder Ž Werner and Pfleiderer ZSK-57.
through 1r4 in. round hole dies at a product
temperature reaching 1458C at the die plate, and pressure ranging between 620 and 740
psi. The extruder screw was 147 cm long with a lengthrdiameter ratio of 24:1. The
material was fed into the conditioner at a rate of 90.9, 93.6 and 113.6 kgrh for whole
peas, dehulled peas and canola meal, respectively, and water was injected at a rate of 4.5
lrh. The material was then dried in a fluid bed dryer at 1108C and pin milled to produce
whole and dehulled extruded meals Ž WEX and DEX, respectively .. Commercial canola
meal was similarly extruded and ground Ž CEX .. Another portion of whole peas was
tempered to 14% moisture and processed using infrared cooking, reaching a temperature
of 1208C. The heat-treated Ž micronized.
peas were subsequently rolled to produce a
flake and then milled into a fine powder Ž WMI .. The proximate composition and effects
of heat treatment on starch gelatinization in the pea meals are presented in Table 1.
Isonitrogenous and isoenergetic Ž gross energy.
experimental diets for the growth trial
were formulated to contain 30% of the test meals, and to meet the nutrient requirements
Ž Akiyama et al., 1991. with proteinrenergy ratios Ž Cruz-Suarez ´ et al., 2000.
recommended
for shrimp Ž Table 2 .. The test pea meals were included in the experimental diets
1 to 5 to replace a portion of a soybean meal and wheat flour mix Ž1:3 parts,
respectively.
in the practical control diet 7. In test diet 6, extruded canola meal replaced
a portion of soybean meal, fish meal and wheat Ž 1:2:3 parts, respectively ..
Table 1
Proximate analysis of the test ingredients, and degree of gelatinization in pea starch before feed manufacture
Ingredients Whole Whole Dehulled Dehulled Whole Canola
peas peas peas peas peas meal
Process raw extruded raw extruded micronized extruded
Ž WRA. Ž WEX. Ž DRA. Ž DEX. Ž WMI. Ž CEX.
Moisture 7.5 8.0 7.9 8.3 6.5 5.7
1
Protein 21.3 21.8 23.7 23.3 21.1 39.4
1
Lipid 1.4 1.6 1.4 1.6 1.5 4.1
1
Ash 3.0 3.0 3.1 3.1 3.2 7.8
1
Crude fiber 6.3 6.5 1.5 1.6 7.2 13.3
1
NFE 67.9 67.2 70.3 70.5 67.1 35.5
2
Gelatinisation 52 389 70 437 66 –
Ž mg glurg.
1 % of the dry matter, NFEsnitrogen free extract, calculated by difference.
2 mg glucose released per g sample by digestion with amyloglucosidase.

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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104
Table 2
Composition of the experimental diets Ž % as fed.
Diets 1 WRA 2 WEX 3 DRA 4 DEX 5 WMI 6 CEX 7 Growth 8 9
control Digestibility diets
Pea Canola
meals meal
1
Soybean meal 7.5 7.5 7.5 7.5 7.5 10 15 10.73 14.35
2
Wheat flour 22.5 22.5 22.5 22.5 22.5 30 45 32.20 43.04
WRA 30
WEX 30
DRA 30
DEX 30
WMI 30
CEX 30
3
Fishmeal 23.26 23.26 23.26 23.26 23.26 13.26 23.26 33.47 19.00
4
Shrimp meal 4 4 4 4 4 4 4 5.74 5.74
Fish oil 1.84 1.84 1.84 1.84 1.84 1.84 1.84 2.65 2.65
Soybean lecithin 4.24 4.24 4.24 4.24 4.24 4.24 4.24 6.08 6.08
Sodium alginate 3 3 3 3 3 3 3 4.31 4.31
Sodium 1 1 1 1 1 1 1 1.43 1.44
hexametaphosphate
5
FP attractant 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.72 0.72
6
Stable vitamin C 0.07 0.07 0.07 0.07 0.07 0.07 0.07 0.1 0.1
7
Mineral mix 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.36 0.36
8
Vitamin mix 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.36 0.36
Choline chloride 0.04 0.04 0.04 0.04 0.04 0.04 0.04 0.06 0.06
Mold inhibitor 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.07 0.07
Antioxidant 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.07 0.07
Ž ethoxiquin.
Chromic oxide 1 1 1 1 1 1 1 1 1
Methionine 0.14 0.14 0.14 0.14 0.14 0.14 0.14 0.21 0.21
Cholesterol 0.31 0.31 0.31 0.31 0.31 0.31 0.31 0.44 0.44
TOTAL 100 100 100 100 100 100 100 100 100
1
Dehulled soybean meal, solvent extracted, 46.3% crude protein as fed Ž CP ..
2
Wheat flour Ž ws330 ., made of 80% hard wheat Ž Canada Western Red Spring principally.
and 20% soft
wheat Ž Canada Eastern Red Wheat 60% and Canada Prairie Spring Red 40% ., 12.3% CP.
3 Chilean jack-mackerel fish meal, 67.6% CP.
4 Chilean pelagic shrimp meal, 39.3% CP.
5
Flavor Pack Ž INVE, Belgium ..
6
Stay-C Ž L-ascorbyl-2-polyphospate, 35% active C ., Roche Vitamins.
7 Mineral mixture composition: Co, 2000 mgrkg; Mn, 16,000 mgrkg; Zn, 40,000 mgrkg; Cu, 20,000
mgrkg; Fe, 1 mgrkg; Se, 100 mgrkg; I, 2000 mgrkg.
8 Vitamin mixture composition: Vit. A, 4000 IUrg; B1, 24,000 mgrkg; B2, 16,000 mgrkg; DL Ca
pantotenate, 30,000 mgrkg; B6, 30,000 mgrkg; B12, 80 mgrkg; C, 60,000 mgrkg; K3, 16,000 mgrkg; D3,
3200 IUrg; E, 60,000 mgrkg; H, 400 mgrkg; niacin, 20,000 mgrkg; folic acid, 4000 mgrkg.
The main ingredients were ground Ž Pulvex 200.
through a a35 screen. The dry
ingredients were mixed and water added to facilitate pelleting through a butchers grinder
Ž Tor-rey.
equipped with 1.6-mm hole diameter die. The pellets were dried in a
convection oven at 1008C for 8 min, allowed to cool and stored at 48C.

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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104 91
2.2. Chemical analysis and water stability tests
Proximate analysis of ingredients and diets were determined according to the
following procedures: crude proteinCP Ž Tecator, 1987 ., moisture ŽA.O.A.C., 1990, No.
929.36 ., ash Ž A.O.A.C., 1990, No. 942.05 ., lipid Ž Tecator, 1983.
and crude fiber
Ž A.O.A.C., 1990, No. 962.09 .. The gelatinization of starch that resulted from the
processing of the peas was measured as glucose released by digestion with amyloglucosidase
Ž Bjork et al., 1987 ..
Water stability tests, replicated five times per diet, were performed with synthetic
marine water at a salinity of 34‰ and temperature between 288C and 298C according to
the method of Aquacop Ž 1978 .. Five-gram samples of the pellets Žspaghetti-like strands,
1.6-mm diameter and approximately 1-cm length.
were gently shaken in wire-mesh
baskets submersed in seawater for 1 h, in order to simulate the conditions on the bottom
of the experimental tanks. The percent dry matter loss Ž %DML.
was calculated as:
%DMLs100= Ž DWdyDWwid.
rDWd; where DWd and DWwid are the dry matter
weights in the diet before and after immersion, respectively. The percent crude protein
loss Ž %CPL. was calculated as follows: %CPLsŽ100=CPdyŽ 100yDML . =
CPwid. rCPd; where CPd and CPwid are the crude protein concentrations Ž% of dry
matter.
in the diet and water immersed diet, respectively.
2.3. Growth trial
The growth trials and digestibility bioassays were conducted at the facilities of the
Programa Maricultura in Monterrey, using a closed recirculating system containing
synthetic marine water with a salinity of 34‰. The shrimp were held in 60-l fiberglass
culture tanks equipped with aeration and constant water temperature control at 288C.
The shrimp for the growth study were selected in a weight range between 200 and
310 mg and distributed, 10 per tank, in four replicated blocks of seven dietary
treatments. A similar weight distribution pattern in each tank was achieved by allotting
the shrimp according to their individual weight. Treatments were randomly assigned to
the tanks within each block. The shrimp were weighed again at 14 and 28 days to obtain
mean body weights for each tank.
Dry feed intake was determined by feeding to satiation. The shrimp were initially fed
a ration of 10% of biomassrday in two feedings daily at 10:00 and 17:00 h. The
following morning leftover feed, which could be readily identified by its swollen pellet
shape, was removed and quantified by estimating the amount in its original dry form,
and the ration adjusted accordingly to minimize the amount of uneaten feed.
The following response variables were determined for each experimental tank: mean
body weight; growth rate expressed as percent weight gain Ž %WG. s100= Žinitial
weightyfinal weight. rŽ initial weight .; dry food intake per shrimp Ž DFI.
was estimated
Ž .
28
from the sum of average daily food intake for each tank DFI sÝ wŽ
is1 intake on ith
day. rŽ number of shrimp on ith day .x; food conversion ratio Ž FCR. sŽfood intake per
shrimp. rŽ average weight gain per shrimp .; protein efficiency ratio Ž PER. sŽweight

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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104
gain. rŽ protein intake .; survival Ž %S. s100= Ž final count. rŽ initial count.
and tank
biomass Ž B. sŽ sum of the individual shrimp weights present in the tank ..
Taking into account the loss of dry matter and protein due to submersion in seawater,
corrected values were calculated multiplying the standard DFI and FCR expressions by
Ž 1y%DMLr100. and dividing the standard PER expression by Ž 1y%CPLr100 ..
2.4. Digestibility determination
The digestibility study was carried out with a separate lot of shrimp obtained from
Aquastrat Ž Sinaloa, Mexico ´ ., which had an average weight of 2.7 g and were distributed,
eight per tank, in two blocks of eight dietary treatments Žsix test dietsqtwo reference
diets .. The collection period was repeated twice to obtain four samples per dietary
treatment, considered as four replicates. The shrimp were fed once a day at a fixed daily
ration of 0.5 grtank for the first two replicates, and 1 grtank for the other two
replicates. After removing uneaten feed, feces were collected at 90, 120 and 150 min
after feeding, for a sufficient number of days Ž 4–8. to pool 1 g feces per sample Žwet
weight .. The feces were collected by siphoning, rinsed immediately with distilled water
and stored frozen. The samples were analyzed for chromic oxide by the method of Bolin
et al. Ž 1952 .. Nitrogen was determined by a modified micro-Kjeldahl method ŽNieto-
Lopez ´ et al., 1997.
in a Tecator equipment using the Bolin reagent. The apparent dry
matter and protein digestibility of diets Ž %ADMD and %APD, respectively.
were
calculated by the equations Ž Maynard et al., 1981 .:
%Crin diet
%ADMDs100y100= %Crin feces
%CPin feces %Crin diet
%APDs100y100= =
%Crin feces %CPin diet
Ž .
where %Cr and %CP are chromium and protein concentrations % of dry matter .
Additionally, apparent digestibility values were adjusted for losses by leaching before
feed ingestion, according to the following equations, in which dry matter or protein
intake is corrected:
%Crin diet 1
%ADMDadj.s100y100= = %Crin feces Ž 1y%DMLr100.
%CPin feces %Crin diet 1
%APDadj.s100y100= = = Ž .
%Crin feces %CPin diet 1y%CPLr100
Apparent digestibility of the ingredient to be tested was determined by the method of
Cho and Slinger Ž 1979 ., where the test ingredient Ž TI.
replaced 30% of the complete
formula of the reference diet Ž RD .. Accordingly, the reference diets Ždiets 8 and 9, Table
2.
were used to calculate the digestibility coefficients of pea meals and canola meal,
respectively, in the test diets Ž TD .. Ingredient’s apparent dry matter and crude protein

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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104 93
digestibility values Ž %IADMD and %IAPD, respectively.
were calculated by the
equations:
Ž 100=%ADMDin TD. yŽŽ 100y%TI . =%ADMDin RD.
%IADMDs
%TI
Ž 100=%APDin TD=%CPin TD. yŽŽ 100y%TI . =%APDin RD=%CPin RD.
%IAPDs
%TI=%CPin TI
where %CP in diet or test ingredient, and the inclusion level of the test ingredient Ž %TI.
are expressed on a dry matter basis.
The apparent digestibility of the nitrogen free extract in diets or ingredients Ž ANFED.
was calculated from the ADMD and APD values, and from assigned values of 90% for
lipids digestibility Ž Cruz-Suarez ´ et al., 1999 ., 50% for ash digestibility Ž Bureau, 2000.
and 0% for crude fiber digestibility:
ANFEDs Ž %drymatter=ADMDy%CP=APDy%lipid=90y%ash=50.
r%NFE
2.5. Metabolizable energy estimation
The metabolizable energy content of the experimental ingredients were calculated by
using the Aaverage productive valuesB proposed by Cuzon and Guillaume Ž 1997.
as
energy coefficients assigned to the digestible energy sources, as follows:
MEkJrgsŽ 17.2= wNFE x=IANFEDadj. . qŽ 39.5= wlipid x=IALD.
qŽ 21.3= wprotein x=IAPDadj.
.
where wNFE x, wlipidx and wproteinx are nitrogen free extract Žassumed to be available
carbohydrate ., crude lipid and crude protein concentrations of the ingredient, and
IANFEDadj., IALD Ž s0.9 ., IAPDadj. are the respective apparent digestibility coefficients.
2.6. Statistical analysis
The responses calculated for each tank Žmean body weight, biomass, growth rate,
survival, food intake, food conversion, protein intake, PER and digestibility.
were
subjected to an analysis of variance and to the Duncan’s multiple range test ŽSteel and
Torrie, 1988.
to first determine whether significant differences existed among the
experimental diets and then to identify where they occurred. In addition, a factorial
analysis of variance was carried out on the raw and extruded, and whole and dehulled
pea meal dietary treatments Ž statistical program SPSS for Windows, release 8.0.0 ..

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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104
3. Results
3.1. Stability of the diets in seawater
Average loss of a dietary dry matter following 1-h immersion was approximately
11% for the diets containing pea meals Ž Table 3 ., except for the whole-extruded pea diet
Ž WEX .. This diet and the reference diets Ž 8 and 9.
showed a better water stability. The
extruded canola diet Ž CEX.
exhibited significantly lower water stability; however crude
protein loss was comparatively low. The WEX diet showed the lowest crude protein loss
while the protein stability of the remaining diets was similar to that of control diet in the
growth trial Ž. 7 . The effects of extrusion on increased starch gelatinization Ž Table 1.
may explain the higher water stability of the diet containing whole-extruded peas.
However, this extrusion effect was not observed for dehulled peas: %DML and CPL
values submitted to factorial ANOVA showed significant probabilities for the rawrextruding
factor Ž Ps0.064 and 0.058, respectively ., but not for the wholerdehulled one
Ž Ps0.112 and 0.254, respectively ..
3.2. Feed intake, growth and surÕiÕal
The growth bioassay results and ANOVA probabilities for the calculated parameters
are presented in Table 4. Highly significant differences Ž P-0.001.
in food intake were
observed. The micronized pea meal diet WMI showed the highest consumption while
both extruded pea meal diets, WEX and DEX showed the lowest intake. The raw pea
Table 3
Proximate analysis and calculated energy content of the experimental diets, and dry matter loss Ž %DML.
and
crude protein loss Ž %CPL.
after 1 h immersion in seawater
1 1 1 1 1 3 3
Diet Moisture Crude Lipid Ash Fiber N.F.E Gross %DML %CPL
1
protein
2
energy
1 WRA 7.5 30.6 9.5 8.7 1.4 42.4 4.3 11.2"0.4 b 19.9"3.0 bc
2 WEX 7.5 29.7 8.9 9.0 2.7 42.3 4.2 9.9"1.0 ab 12.6"1.1 a
3 DRA 7.6 30.5 9.6 8.6 0.9 42.8 4.4 11.3"1.0 bc 17.4"3.1 b
4 DEX 7.3 30.8 9.3 8.6 0.9 43.1 4.4 11.1"0.9 b 18.5"1.9 b
5 WMI 7.0 29.6 9.6 8.6 2.4 42.9 4.3 11.2"1.4 b 18.9"1.8 b
6 CEX 7.2 30.2 9.3 8.8 3.8 40.7 4.2 12.6"0.8 c 17.2"1.4 b
7 Control 6.8 31.2 8.9 8.5 1.5 43.2 4.4 11.1"0.5 b 18.9"0.4 b
growth
8 Reference 6.6 35.8 12.8 11.0 1.5 32.3 4.5 9.6"1.0 a 17.3"0.8 b
pea meal
9 Reference 6.7 29.0 11.8 9.1 1.6 41.9 4.5 9.6"0.7 a 19.1"0.6 b
canola
1 % as fed. NFEsnitrogen free extract, calculated by difference.
2
Proteins5.6 kcalrg, lipids9.5 kcalrg, carbohydratess4.1 kcalrg Ž Tacon, 1987 ..
3 Different letters in same column indicate different homogeneous subsets as determined by Duncan’s
multiple comparisons Ž P s0.05 .. One-way ANOVA probabilities were P s0.001 and P s0.006 for DML
and CPL data, respectively.

Table 4
Growth experiment results Ž means of four replicates values"standard deviation.
Treatments 1 WRA 2 WEX 3 DRA 4 DEX 5 WMI 6 CEX 7 Control Prob. 1
Mean body weight
Initial Ž mg.
255"3 254"2 257"2 255"1 256"3 255"3 256"2 0.497
14 days Ž. g 0.61"0.03 abc 0.57"0.02 a 0.60"0.03 ab 0.61"0.02 abc 0.65"0.03 c 0.60"0.05 ab 0.62"0.03 bc 0.040
28 days Ž. g 1.03"0.11 a 0.95"0.03 a 0.98"0.06 a 0.99"0.07 a 1.13"0.07 b 0.94"0.05 a 1.01"0.04 a 0.012
Dry feed intake Ž. g
28 days 1.18"0.10 b 1.03"0.03 a 1.28"0.07 b 1.07"0.04 a 1.56"0.05 c 1.22"0.04 b 1.29"0.09 b -0.001
Dry feed intake, corrected for losses by leaching before ingestion Ž. g
28 days 1.05"0.01 b 0.93"0.02 a 1.13"0.06 b 0.95"0.04 a 1.38"0.04 c 1.07"0.04 b 1.14"0.08 b -0.001
Ž .
Weight gain %WG
28 days 301"40 a 273"10 a 282"23 a 288"25 a 340"29 b 269"18 a 292"14 a 0.011
Feed conversion ratio
28 days 1.6"0.1 ab 1.5"0.1 a 1.8"0.2 c 1.5"0.1 a 1.8"0.2 c 1.8"0.1 c 1.7"0.05 bc -0.001
Feed conversion ratio, corrected for losses by leaching before ingestion
28 days 1.4"0.1 ab 1.3"0.07 a 1.6"0.15 c 1.3"0.07 a 1.6"0.14 c 1.6"0.04 c 1.5"0.04 bc 0.001
Protein efficiency ratio
28 days 2.0"0.2 b 2.1"0.1 b 1.7"0.2 a 2.1"0.1 b 1.8"0.1 a 1.7"0.1 a 1.8"0.05 a -0.001
Protein efficiency ratio, corrected for losses by leaching before ingestion
28 days 2.5"0.2 b 2.4"0.1 b 2.1"0.2 a 2.5"0.1 b 2.2"0.2 a 2.1"0.1 a 2.2"0.06 a 0.001
Survival Ž %.
28 days 93"10 90"8 95"6 98"5 100"0 90"0 100"0 0.061
Biomass Ž. g
28 days 9.5"1.7 ab 8.5"0.7 a 9.3"0.4 ab 9.6"1 ab 11.2"0.7 c 8.4"0.5 a 10.1"0.4 bc 0.003
1 Prob.sProbability by a one-way analysis of variance between treatments. Treatment means within the same row with different letters are significantly different
Ž .
Duncan’s test, P s0.05 .
L.E. Cruz-Suarez et al.rAquaculture 196 ( 2001 ) 87–104 95

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meal Ž WRA and DRA ., the extruded canola meal Ž CEX.
and control diets showed
intermediate consumption. Weight gain of the experimental animals practically tripled
after 28 days for each of the dietary treatments. With the exception of the micronized
pea Ž WMI dietary treatment ., there were no significant differences in weight gain among
the dietary treatments. A food conversion ratio of less than 2 was obtained for each of
the dietary treatments. Significantly lower feed conversion ratios were obtained for the
extruded pea meal diets, perhaps due to improved starch digestibility. The highest PER
values were also obtained for the extruded pea meal diets indicating that protein
utilization was not impaired by the effects of heat treatment. Correction of DFI for
losses in sea water before the feed ingestion led to improved values of FCR and PER,
but relative differences among the means for the different treatments were not modified
substantially, since homogeneous subsets as defined by the Duncan’s multiple comparison
test remained the same. Survival for all treatments Ž above 90% . was excellent and
did not differ significantly amongst diets. The largest biomass gain was obtained for
shrimp fed the WMI micronized pea and control diet, due mainly to the weight gains of
these groups.
3.3. Digestibility
The apparent dry matter digestibility Ž ADMD.
for the experimental diets was
generally higher than that of each of the reference diets Ž Tables 5 and 6 .. The ADMD
and apparent protein digestibility Ž APD.
amongst the experimental growth study diets
Ž 1–6.
were not significantly different. Apparent dry matter and protein digestibility
values for the test ingredients calculated using reference diets Ž 8 and 9.
and following
the method of Cho and Slinger Ž 1979.
are also presented in Tables 5 and 6. The
adjustment of the digestibility coefficients for leaching losses Ž Table 6 vs. Table 5.
did
not change the statistical inference to the treatment effects. However, in the subsequent
calculation of the carbohydrate digestibility, it was noted that the adjusted coefficients
supported more realistic values that would indicate the correction to be necessary,
especially for the extruded canola meal Ž Table 5 .. The highest dry matter digestibility
values were obtained with the extruded pea flours and the lowest with micronized pea
meal and extruded canola meal. Similarly, adjusted digestibility of carbohydrate as
expressed by NFE Ž Table 6.
was close to 100% for the pea ingredients and canola
except for DRA and WMI Ž 87.4% and 85.5% .. In contrast, there were no significant
differences in IAPD among the differently processed pea meals and canola meal.
However, in spite of these relatively uniform protein digestibility values, and lower
carbohydrate digestibility for the micronized pea, the feeding trial results show best
performance with the WMI treatment. Therefore, the benefit derived from micronizing
peas is apparently due to enhanced feed intake rather than an improvement in nutritional
value.
Metabolizable energy calculations led to higher values for extruded pea meals
Ž 16.2–16.4 kJrg. than for raw pea meals Ž 15.4–15.6 kJrg ., and lowest values for
micronized pea and canola Ž 14.0 and 14.2 kJrg, respectively. Ž Table 6 .. Metabolizable
energy was lower than digestible energy by only about 0.4 kJrg, for all the experimental
ingredients.

Table 5
Apparent dry matter, protein and NFE digestibility in diets and ingredients, as obtained by standard determination Ž mean of four replicate values"standard deviation.
1 WRA 2 WEX 3 DRA 4 DEX 5 WMI 6 CEX 8 Ref. pea 9 Ref. Canola ANOV
Prob.
Diet digestibility (%), standard
ADMD 79.3"1.8 bc 79.9"1.2 c 78.8"1.9 bc 80.2"0.9 c 76.8"2.7 abc 77.3"3.2 abc 75.1"2.2 a 76.4"2.1 ab 0.020
APD 89.8"0.9 ab 90.6"1.1 b 91.0"1.2 b 90.4"0.7 ab 90.1"2.2 ab 91.3"2.0 b 88.0"2.3 a 90.9"1.0 b 0.123
ANFED 77.8"3.4 cd 81.7"2.0 d 74.9"3.6 bc 78.6"1.8 cd 74.1"4.2 bc 77.2"5.8 cd 68.9"4.2 a 71.2"4.0 ab 0.001
Ingredient digestibility (%), standard
IADMD 89.0"6.2 abc 91.3"3.9 bc 87.6"6.5 abc 92.4"2.9 c 80.7"8.8 ab 79.4"10.6 a 0.075
IAPD 79.8"4.6 83.0"5.3 85.4"5.8 82.7"3.2 80.0"11.2 80.0"5.4 0.707
IANFED 102.3"7.8 104.4"4.5 91.8"7.8 99.6"4.0 90.7"9.7 113.6"23.8 0.293
Ž .
Different letters in same raw indicate different homogeneous subsets Duncan 0.05% .
L.E. Cruz-Suarez et al.rAquaculture 196 ( 2001 ) 87–104 97

98
Table 6
Apparent dry matter, protein and NFE digestibility in diets and ingredients, adjusted for losses by leaching before ingestion, and corresponding energy contents in pea
and canola meals Ž mean of four replicate values"standard deviation.
1 WRA 2 WEX 3 DRA 4 DEX 5 WMI 6 CEX 8 Ref. pea 9 Ref. Canola ANOV
Prob.
Diet digestibility (%), adjusted
ADMD 76.6"2.1 b 77.7"1.3 b 76.1"2.2 ab 77.8"1.0 b 73.8"3.0 ab 74.1"3.6 ab 72.5"2.5 a 73.9"2.3 ab 0.027
APD 87.3"1.1 ab 89.3"1.2 b 89.1"1.5 b 88.2"0.8 ab 87.7"2.7 ab 89.5"2.4 b 85.5"2.8 a 88.8"1.3 b 0.09
ANFED 73.9"3.8 cd 77.8"2.2 d 70.4"4.0 bc 74.9"2.0 cd 69.4"4.6 bc 71.1"6.6 bcd 62.1"4.5 a 67.0"4.3 ab 0.09
Ingredient digestibility (%), adjusted
IADMD 86.5"6.9 ab 90.1"4.4 b 84.7"7.4 ab 90.4"3.3 b 77.0"10.0 a 74.4"12.1 a 0.045
IAPD 77.1"5.7 87.2"6.1 86.3"7.0 82.2"3.9 79.0"13.8 79.0"6.5 0.371
IANFED 99.1"8.6 101.4"5.0 87.4"8.9 96.8"4.5 85.5"10.5 100.5"26.9 0.671
Ingredient energy content ( kJr g)
1
ME 15.6"1.2
2
16.2"0.8 15.4"1.3 16.4"0.6 14.0"1.8
2
14.2"2.2 0.307
3
DE 15.9"1.3
2
16.6"0.8 15.8"1.3 16.8"0.6 14.3"1.9
2
14.8"2.2 0.383
Different letters in same raw indicate different homogeneous subsets Ž Duncan 0.05% ..
1
Metabolizable energy was calculated as described in material and methods by assigning average productive values Ž Cuzon and Guillaume, 1997.
to the digestible
nutrient contents as calculated with adjusted digestibility coefficients for protein and NFE.
2 Although IANFEDadj. was calculated at 101.2% for WEX and 100.5 for CEX, we used a 100% value for the calculations of metabolizable and digestible energy
contents.
3
Digestible energy was calculated by assigning gross energy values Ž Tacon, 1987. to the digestible nutrient contents, adjusted for leaching: DE kJrgsŽ23.4=
wprotein x =IAPDadj. . qŽ39.7= wlipid x =0.90. qŽ17.2= wNFE x =IANFEDadj. ..
L.E. Cruz-Suarez et al.rAquaculture 196 ( 2001 ) 87–104

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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104 99
3.4. Analysis of the effects of dehulling and extrusion
Factorial ANOVA of the results for diets 1, 2, 3 and 4 showed that the effects from
extrusion were generally higher than from dehulling. No significant effects on performance
were observed as a result of dehulling, although a higher feed consumption and
lower PER approached significance Ž Ps0.121 and Ps0.073, respectively ..
Extrusion reduced feed intake Ž P-0.001 ., improved feed conversion Ž Ps0.009 .,
increased PER Ž Ps0.006.
and tended to improve dry matter digestibility of peas
Ž Ps0.192 .. Extrusion had no effect on growth Ž Ps0.44 ., survival Ž Ps1.
and protein
digestibility Ž Ps0.751 .. These results again can be explained by improved digestibility
of gelatinized over native starch.
4. Discussion
The impact of dry matter and protein losses due to leaching before ingestion has been
evaluated through the calculation of corrected values, which take into account the losses
that occurred in the diets after 1 h. This simple correction seems justified by considering
that most of the losses occur during the first hour of immersion ŽRomero-Alvarez,
1995 ., and that the average interval time of feed consumed was generally noted to be
approximately 1 h after offering the feed. Corrected values showed the same relative
positions as standard values, even in the case of the WEX treatment, which stability in
water was significantly better than that of other diets. The corrected values are of
interest because they indicate the true biological values of feed intake, conversion ratio
and PER as a more accurate measurement of the crustacean’s metabolic response to the
dietary treatments. They also allowed a truer estimation of the carbohydrate digestibility
of the experimental ingredients. The uncorrected values on the other hand reflect the
results of a practical significance, which can be compared with other studies using
standard calculations.
The results of this study show that raw peas, and peas processed by dehulling,
extruding and micronizing, have a nutritional value similar to that of a mixture Ž 1:3.
of
soybean meal and wheat flour Ž 80% hard–20% soft varieties, Table 2.
in shrimp diets.
Also, substituting a portion of the soybean meal, fish meal and wheat Ž1:2:3 parts,
respectively.
with extruded canola meal did not affect shrimp performance. Our results
corroborate the reports by other authors cited in the introduction that pea seed meal and
canola meal can be successfully included in practical aquaculture feeds. When alternate
plant origin ingredients are used in diets containing the same concentration of digestible
energy and digestible protein, and meeting nutrient requirements, similar performance
can be expected. However, Gomes et al. Ž 1995.
found that voluntary food intake of trout
fed diets including coextruded peas and canola was depressed due to inherent factors
present in the plant ingredients Ž Novoa and Castillo, 1998 .. Therefore, similar growth
was not obtained. In the present study, no adverse response to raw whole peas was
observed. Studies with trout Ž Kaushik et al., 1993. and European sea bass ŽGouveia and
Davies, 2000.
have emphasized the requirement for dehulling and extrusion cooking
under conditions that would be more severe than they would during complete feed

100
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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104
manufacture. Allan Ž 1997.
cites unpublished data that show an improvement in the
nutritional value of Dunn peas Ž a tannin containing P. satiÕum variety.
through
dehulling and extrusion when included in the diets of silver perch, an omnivorous fish.
In the present study, no particular benefit was obtained by dehulling the pea, perhaps
because the field pea strains used to produce the experimental meals have been
improved. Eusebio Ž 1991. also found that dehulling cowpeas and rice beans Žleguminous
seeds.
had no significant effect on growth and survival of Pen. monodon although
apparent protein digestibility of rice beans but not cowpeas was improved. Shrimp may
actually be more tolerant of certain anti-nutritional seed coat compounds than other
species.
Smith et al. Ž 1999.
reported digestibility coefficients of field peas in tiger shrimp of
80% for IADMD and 91% for IAPD in tiger shrimp. The IADMD value is lower and the
IAPD value is higher than those obtained in present study Ž88–92% and 79–85%,
respectively.Ž Table 5 .. The lower feed consumption of the extruded pea diets did not
reduce growth, suggesting higher energy availability from these diets ŽCuzon and
Guillaume, 1997 .. The higher dry matter digestibility of extruded peas also suggests a
higher digestible energy value for gelatinized starch resulting from the effects of
extrusion cooking. Davis and Arnold Ž 1993.
reported that the gelatinization of starch in
sorghum significantly improved digestibility in Pen. Õannamei. However, Davis and
Arnold Ž 1995.
found that starch gelatinization did not necessarily correspond to increased
energy digestibility of the cereal grains wheat and rice as observed for corn and
sorghum. The different responses were attributed to variable effects of wet and dry
extrusion conditions with different grains. In the present study gelatinization of starch by
wet extrusion had a small positive effect on digestibility and performance, although the
raw pea meal was quite acceptable and perhaps the extrusion costs would not be
recovered.
Cousin et al. Ž 1996.
also found differences in native and gelatinized starch digestibility
measured both in vivo and in vitro depending on the source of dietary starch with
reference to the amyloseramylopectin composition and its intrinsic starch granule
characteristics. The proportion of amylose ranges between 23.5% and 35% in feed pea
starch compared to over 66% in wrinkled canning type peas Ž Castell et al., 1996 ..
According to Grosjean et al. Ž 1998.
lower apparent energy digestibility in pigs of
wrinkled compared to feed pea is partly due to the amyloseramylopectin ratio. The data
reported by Cousin et al. Ž 1996.
comparing high amylopectin and high amylose corn
starch and wheat starch by in vivo and in vitro assay with L. Õannamei also showed
lower starch digestibility from the high amylose source. The feed peas used in this study
can be considered as having an amylose content closer to that of wheat starch Ž 23% .,
which is also highly digestible in both native and gelatinized forms ŽCruz-Suarez ´ et al.,
1994; Davis and Arnold, 1995; Cousin et al., 1996 ..
Infrared heat treatment did not increase starch gelatinization and digestibility, although
feed intake and growth was considerably higher and food conversion was good
with the micronized pea diet. The higher growth was approximately 16% higher than the
control and 20% higher than the raw and extruded pea fed groups. Therefore, the
beneficial effects of micronization may have resulted from the development of compounds
that enhanced palatability andror the elimination of off flavors for blue shrimp.

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L.E. Cruz-Suarez et al.rAquaculture 196 2001 87–104 101
Gomes et al. Ž 1995.
reported a 10% increase in protein and energy digestibility from
micronizing full fat soybeans and a 21% increase in energy digestibility, but not protein,
from micronizing wheat fed to rainbow trout. The infrared heat treatment and low
moisture tempering were perhaps not severe enough to improve starch digestibility in
the present study.
Metabolizable energy contents in peas for P. stylirostris were higher than those for
swine and poultry Ž 13.6 and 11.1 kJrg according to Castell et al., 1996 .. Higher
metabolizable energy values in shrimp than in mammalian and avian species are in
accord with their mode of excreting nitrogen in form of ammonium ŽCuzon and
Guillaume, 1997 .. Metabolizable energy content in peas increased from 15.4 to 16.4
kJrg by extruding the pea meal most likely due to the improved digestibility of
gelatinized starch.
Dry matter and protein digestibility of the extruded canola diet was found to be very
good and similar to that of a practical diet formulation that included a high quality fish
meal. These results agree with those of Buchanan et al. Ž 1997.
who reported on the
successful use of canola meal in Pen. monodon diets at both a 20% inclusion level, and
64% inclusion with the addition an enzyme mixture. Satoh et al. Ž 1998.
reported that
extrusion processing of canola meal improved its nutritive value as measured by growth
of chinook salmon in sea water. Protein and lipid contents were increased and phytic
acid and glucosinolates, that interfere with nutrient bioavailability and thyroid function,
respectively, were partially degraded. The results of this study suggest that extruded feed
peas and canola would complement each other in practical shrimp diets. Co-extruded
rapeseed and peas have replaced up to 66% of brown fish meals in rainbow trout diets
Ž Gomes et al., 1995 ..
This study demonstrates the acceptable nutritional value of peas and canola meal as
ingredients in diets for the production of shrimp, since they could replace common
shrimp feed ingredients. Inclusion of peas and canola meal in shrimp diets will therefore
be a function of diet formulation and commodity prices. The effects of the micronizing
process on pea meals deserve more research to investigate how the process could
increase feed consumption more than that required to compensate for reduced dietary
energy availability.
Acknowledgements
We thank shrimp farms Cristo Rey and Aquastrat, Sinaloa, Mexico for providing
´
juvenile Litopenaeus stylirostris for the growth and digestibility experiments, respectively,
and Servando Quiroz B., Claudio Guajardo B. and Norma E. Luna for their help
in formulating, analyzing and testing the shrimp feeds. The peas were provided by Paul
Adelizi of the Saskatchewan Wheat Pool. We express our gratitude to Francis and Tony
Gaudet of Belle Pulses for dehulling peas, Connie Phillips and Kevin Swallow of the
Alberta Food Processing Centre, Leduc, for extruding the meals and Mark Pickard and
Gordon Sellar of Infraready Products, Saskatoon, for micronizing the peas. We thank the
Saskatchewan Pulse Growers and the Canadian International Grains Institute for funding
this study.

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