Utilization of Feed Peas (Pisum Sativum)

8/11/2019 Utilization of Feed Peas (Pisum Sativum)

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Final Project Report

Title: Utilization of feed peas (Pisum sativum) as alternative protein sources in diets forshrimp, tilapia, and milkfish.

Implementing Personnel and Agencies:

Southeast Asian Fisheries Development Center, Aquaculture DepartmentClarissa L. Marte, PhD, Head, SEAFDEC Research Division ([email protected])Myrna B. Teruel, Study leader, Shrimp ([email protected])Corazon B. Santiago, PhD, Study leader, Tilapia ([email protected])

Ilda G. Borlongan, Study leader, Milkfish ([email protected])Perla S. Eusebio, Study leader, Digestibility experiments ([email protected])

USA Dry Pea and Lentil CouncilTimothy Welsh, Southeast Asia Representative ([email protected])

Funding: US Department of Agriculture, Emerging Markets ProgramUSA Dry Pea and Lentil Council

Trial Period: July, 2001 through March, 2002


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Table of Contents

SUMMARY ............................................................................................................................................................1-2

STUDY 1. SHRIMP (PENAEUS MONODON, FABRICIUS) ............................................................................................... 1

STUDY 2. TILAPIA (OREOCHROMIS NILOTICUS

L.)..................................................................................................... 1STUDY 3. MILKFISH (CHANOS CHANOS, FORSSKAL) ................................................................................................. 2

GENERAL CONCLUSIONS ............................................................................................................................................ 2

Utilization Of Feed Pea, Pisum Sativum Meal, As An Alternative Protein Source

In Practical Diets For Juvenile Tiger Shrimp, Penaeus Monodon Fabricius......................................................3-10

ABSTRACT................................................................................................................................................................... 3

INTRODUCTION............................................................................................................................................................ 3

MATERIALS AND METHODS........................................................................................................................................ 4

RESULTS ...................................................................................................................................................................... 5

DISCUSSION................................................................................................................................................................. 5

ACKNOWLEDGEMENT................................................................................................................................................. 9

REFERENCES: .............................................................................................................................................................. 9

Feed Pea (Pisum Sativum) As An Alternative Protein SourceIn The Diets Of Nile Tilapia, Oreochromis Niloticus(L.) ................................................................................11-19

ABSTRACT................................................................................................................................................................. 11

INTRODUCTION.......................................................................................................................................................... 11

MATERIALS AND METHODS...................................................................................................................................... 11

RESULTS .................................................................................................................................................................... 13

DISCUSSION............................................................................................................................................................... 14

ACKNOWLEDGEMENTS ............................................................................................................................................. 18

REFERENCES.............................................................................................................................................................. 18

Potential Of Feed Pea (Pisum Sativum) As An Alternative Protein Source

In Practical Diets For Milkfish (Chanos Chanos Forsskal)...............................................................................20-26

ABSTRACT................................................................................................................................................................. 20

INTRODUCTION.......................................................................................................................................................... 20MATERIALS AND METHODS...................................................................................................................................... 20

RESULTS .................................................................................................................................................................... 22

DISCUSSION............................................................................................................................................................... 22

ACKNOWLEDGEMENT............................................................................................................................................... 26

REFERENCES.............................................................................................................................................................. 26


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Digestibility Experiments. As ingredient in tilapia diet, feed pea has apparent dry matter digestibility(ADMD) of 69.3+1.8% and apparent protein digestibility (APD) of 87.22.0%. The digestibility of some ofthe diets used in experiments 1 and 2 was also determined in separate feeding trials. The ADMD of thediets increased as feed pea inclusion level increased. The APD of the fishmeal-based diets or the plant-based diets were all high, exceeding 90%.

Overall, feed pea meal may substitute up to 50% of fish meal protein (equivalent to 63.3% feed pea) and35% of the plant proteins (41% feed pea) in the diets of Nile tilapia without any adverse effect on fish

growth and survival.

Study 3. Milkfish (Chanos chanos, Forsskal)The efficacy of feed pea meal as an alternative protein source for milkfish was tested in a 12-week feedingtrial. All test diets were made isonitrogenous (30% crude protein) and isocaloric (15.7 kJ g

1). The control

diet contained fish meal, soybean meal, meat and bone meal, and copra meal as protein sources. Feedpea meal was substituted at 0, 5, 10, 15, 20, 25, and 30% of total protein. A leading commercial milkfishfeed was also tested as an additional control. The experimental diets were fed to triplicate groups ofmilkfish fingerlings (mean initial weight =0.420.01g) at 10% body weight /day.

Results showed that % weight gain (WG) of milkfish juveniles fed diets with 5% (WG= 833.1) and 10%(WG=835.3) feed pea inclusion did not significantly differ from that of the SEAFDEC control diet (0% feed

pea inclusion; WG=834.8). However, % weight gain obtained for milkfish juveniles fed with 15%(WG=691.0) and 20% (WG=680.0) feed pea inclusion in the diet were significantly higher than thecommercial milkfish feed formulation (no feed pea inclusion, WG=581.4). Survival ranged from 70-90%.FCR was between 2.1 to 2.60 while PER ranged from 1.15 to 1.44.

Dietary crude protein digestibility values of the experimental diets were 84.8 to 89.2% suggesting that feedpea protein is well digested by milkfish.

Over-all results indicate that feed peas could be used as an alternate dietary protein source for milkfishjuveniles. Optimum level of inclusion without amino acid supplementation is 10% of the dietary protein(131g feed pea/kg dry diet). Dietary protein inclusion levels up to 20% feed peas (260g/kg diet) are stillacceptable.

General ConclusionsThese feeding trials clearly demonstrate that whole feed pea can be incorporated as an alternative proteinsource in diets for tilapia, milkfish, and shrimp. Since feed peas have a moderate protein content (23-25%)compared to fish meal (65-70%), their incorporation in fish diets needs to be balanced with other proteiningredients, especially for fish that require high protein levels in their diet. In this case, feed pea cantherefore be used as one of the ingredients that would collectively replace fish meal in practical diets.

The replacement of other plant protein sources with feed pea is possible as evidenced by the studiesconducted. Feed peas contain high levels of lysine, which could complement the amino acids of otheringredient substitutes that are deficient in such amino acids. The relatively low level of sulfur amino acids(i.e., methionine and cystine) in feed pea meal may limit the use of this plant protein source dependingupon the requirement of the species and the diet formulation.

The pea variety used in these studies contains low levels of anti-nutritional factors and is similar to mostfeed pea varieties now supplied in global markets. Thus, positive results were obtained for shrimp andtilapia, even at high levels of incorporation in the diets. The relatively lower performance of milkfishcompared to shrimp and tilapia, even with feed pea comprising 15% of the diet, may be attributed to aminoacid deficiencies or possibly to effects from certain anti-nutritional factors. However, it is notable that thegrowth response of milkfish fed the 15% feed pea diet was still better than from the commercial milkfishdiet, which contained no feed pea, so further study is needed.

The availability of feed pea meal as an alternative protein source for tilapia, milkfish, and shrimp may openavenues for commercial producers to reduce feed costs. Feed peas have a higher carbohydrate content(65-67%) compared to soybean meal (44-45%), which may provide a lower cost and more digestible energysource for the animals.


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Utilization Of Feed Pea, Pisum Sativum Meal, As An Alternative Protein SourceIn Practical Diets For Juvenile Tiger Shrimp, Penaeus MonodonFabricius

Myrna N. Bautista-Teruel, Perla S. Eusebio, and Timothy P. Welsh

AbstractThe potential of feed pea, Pisum sativum, meal as an alternative protein source in practical diets for the

juvenile tiger shrimp, Penaeus monodon, was assessed in several experiments. Six isonitrogenous diets

were formulated to contain 40% protein. Protein from feed pea meal replaced 0, 20, 40, 60, 80, and 100%of the protein from defatted soybean meal in the diets. These values were equivalent to 0, 5, 10, 15, 20,and 25% respectively of the total protein in the diet. A negative control with no protein sources was addedto the treatments. Twelve shrimp post-larvae with an average weight of 0.020.01g were randomlyassigned in 35, 60-l oval tanks equipped with a flow-through seawater system. The shrimps were fed theformulated diets at a daily feeding rate of 20-25% body weight for 90 days in 5 replicate samples. Nosignificant differences (P0.05) compared to the rest of thetreatments. Specific growth rates (SGR) of shrimps for the various treatments showed the same trend asthe percent weight gain. Survival of shrimp for all treatments, including the negative control, was generallyhigh at 75-100%. The apparent dry matter (ADMD) and protein (APD) digestibilities of the dry feed pea in P.monodonwere high at 73.384.98 and 92.742.62, respectively. Digestibility coefficients for dry matter and

protein for the feed pea meal-based diets increased with increasing level of feed pea replacement. Therewere no significant differences (P


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Materials and MethodsExperimental diets

The feed ingredients used in this study were obtained from commercial suppliers through the SEAFDECpilot feed mill plant. Feed pea of US origin was supplied through a local distributor in Manila, Philippines.Prior to use, all dry feed ingredients were ground and sieved through a 60m mesh screen sieve.Proximate analyses for each feed ingredient were conducted and the resulting data were used in dietformulation (Table 1). Six isonitrogenous diets were formulated to contain 40% protein (Table 2). Proteinfrom feed pea replaced 0, 20, 40, 60, 80, and 100% of the protein from defatted soybean meal in the diets.These values were equivalent to 0, 5, 10, 15, 20, 25%, respectively, of the total protein in the diet.Replacement of protein was based on the essential amino acid requirements of tiger shrimp (Kanazawaand Teshima, 1981; NRC, 1993; FDS Manual, 1994; Millamena et al., 1996-1999). A negative control withno protein sources was added to the treatments. Protein content of 40% in the shrimp diet has beenreported to be optimal for juvenilePenaeus monodon (Alava and Lim 1983; Bautista, 1986; Shiau et al.1991). The diet containing no feed pea meal served as the control diet. Other animal protein sources, suchas fish meal, shrimp meal, shrimp head meal, and squid meal, were included in the dietary formulations atthe same levels. The levels of wheat flour and rice bran were adjusted to maintain similar dietary proteincontents. Diets were prepared according to standard diet preparation in the laboratory as described in FDSManual, 1994. To prevent lipid oxidation during storage, all diets were packed in sealed plastic containersand stored inside a cold room at 20C until used in the feeding trials.

Proximate analyses were conducted of the various diets prepared following the methods of Association ofOfficial Analytical Chemists (AOAC 1990). The amino acid composition of both soybean meal and feed peameal and experimental diets was determined using the method of Simpson et al., 1976.

Feeding trials

Good quality and disease-free post-larvalPenaeus monodon were sourced from a private hatchery inTigbauan, Iloilo, Philippines. They were placed in 2-tonne fiberglass tanks and fed withArtemia sp andcontrol diet for a week before they were transferred to experimental tanks. Twelve shrimp post-larvae withan average weight of 0.02 0.01g were randomly assigned in 35, 60 L oval tanks equipped with a flow-through seawater system. Prior to start of the feeding trial, shrimps were acclimatized to experimental diets.The shrimp were fed the formulated diets at a daily feeding rate of 20-25% of total body weight for 90 daysin 5 replicate samples. Shrimp were fed three times daily at 0800, 1300, and 1700h. The wet weights ofthe shrimp were recorded every 15-d interval for 90 days and the amount of feed given was adjusted every

sampling day. All experimental tanks were cleaned before every feeding. Water temperature, salinity,dissolved oxygen, and pH were measured daily in all tanks. Total values for ammonia-nitrogen and nitrite-nitrogen were measured once a week. Shrimp were collected at the beginning of the feeding trial forproximate analyses. At the conclusion of the feeding trial, all shrimp samples from each tank were pooled,freeze-dried and ground for proximate analyses of whole body composition following the standard methodsof AOAC (1990).

Biological analysisDiet performance was evaluated by calculation of percent weight gain=final weight-initial weight/initialweight x 100; specific growth rate=100 (ln ave. final wt-ln ave. initial wt)/no. of culture days; feed conversionratio (FCR)= total dry feed intake (g)/wet weight gain (g); percent survival =final number of shrimp/initialnumber of shrimp x 100.

In vivo digestibility experimentApparent digestibility coefficients for dry matter (ADMD) and crude protein (APD) of feed peas as ingredientand feed-peas based diets were measured using 1% Cr2O3 as external indicator. The method by Cho. etal. (1982) was adapted in a ratio of 70:30 (reference diet to test ingredient) in each test diet. The controldiet used for growth experiment was used as reference diet. Composition of the diet is shown in Table 3.

In the digestibility experiment for the test ingredient and feed pea-based diets, 10 shrimp with mean bodyweight of 14.53 0.57g and 8 shrimp with mean body weight of 13.560.56g were stocked in 250 L conicalfiberglass tanks for the test ingredient and feed pea-based diets, respectively. The shrimp were acclimatedto experimental diets and laboratory conditions for 7 days before the experiment. All shrimp were fed at 8-10% of the total body weight 3x daily. A flow-through filtered seawater system with continuous aeration andflow rate of 600-700 ml/min was provided for each tank. Water temperature and salinity were maintained at24-48C and at 30-34 ppt, respectively. Culture period was 80 days. Fecal collection was done manually.

The feces were allowed to float into a plastic scoop and then pipetted and gently transferred into acollecting vial. Care was taken to prevent the breaking-up of fecal strands to facilitate collection and toavoid leaching of nutrients. Thereafter, fecal samples were carefully rinsed with distilled water andimmediately stored in a freezer to retard bacterial decomposition. At the end of fecal collection, samples


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were thawed and pooled prior to freeze-drying. Dried fecal samples were thoroughly mixed and preparedfor chromic (Cr2O3) (Carter et al., 1960) and crude protein analysis (AOAC, 1990). Apparent dry matterdigestibility (ADMD) and apparent protein digestibility (APD) were determined using the equation:

% Digestibility = 100-100 X [{Cd/Cf} X {Nf/Nd}] where: Cd= % chromic oxide in diet; Cf= % chromic oxide infeces; Nf= % nutrient in feces; Nd= % nutrient in diet

Water stability test for dietsWater stability of the diets was determined at 4, 8, 12, 16, and 24 h following the method described in FDSManual, 1994. Percent water stability was computed as final dry weight of feed / initial dry weight of feed X100.

Statistical analysis

All data were analyzed by a one-way ANOVA using a Statistical Analysis Software Program of SPSS. TheDuncans Multiple Comparisons T was used to determine the differences between the treatment means(Duncan, 1955). Results were considered statistically significant at the level of P


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Smith et al. (1999) reported that digestibility coefficients of feed peas in tiger shrimp were 80% for ADMDand 91% for APD. The ADMD value and APD value is almost the same as those obtained in this presentstudy (73% and 89-92%).

In this study, no adverse effect on shrimp was noted on the use of whole unprocessed feed pea. Perhaps,processing of feed pea may even improve its present nutritional value. Cruz-Suarez et al., 2001 however,did not obtain any particular benefit with the use of processed feed pea (dehulling). In her study, raw feedpeas and peas processed by dehulling have similar nutritional value in shrimp diets. Results, however, may

be dependent on the type or variety of feed pea used. Studies with other species such as trout (Kaushik etal., 1993) and European sea bass (Giuveia and Davies, 2000) have emphasized the requirement fordehulling and extrusion cooking. Eusebio (1991) also found that processing of cowpea and rice bean hadno significant effect on growth and survival of P. monodon,although apparent protein digestibility of ricebean was improved. Shrimp are more tolerant than other species of certain anti-nutritional factors in theseed coat components (Cruz-Suarez et al., 2001).

This study has demonstrated the acceptable nutritional value of whole feed pea as an ingredient in shrimpdiets, since this product can replace most commonly used shrimp feed ingredients, such as soybean meal.Inclusion of feed peas in shrimp diets may therefore be a function of diet formulation. An inclusion level upto 42% in the juvenile shrimp, P. monodon, practical diet does not give any adverse effect on growth,survival, and body composition of the animal.

Table 1 Proximate composition (% dry weight) of feed ingredients included in test diets*

Table 2 Percentage composition of experimental diets on a dry matter basis (g/100g dry diet)

Ingredients MoistureCrude

Protein(N x 6.25)

Crude fatCrudefiber

NFEc Ash

Feed peaa

11.58 25.24 1.32 3.68 65.96 3.80

Soybean mealb

10.78 42.67 1.37 4.03 44.87 7.06

Seaweed 9.1 12.13 0.44 5.58 30.42 51.43

Wheat flour 14.19 17.21 1.26 0.03 80.76 0.74

Squid meal 15.34 78.84 5.19 0.55 5.38 10.04

Shrimp meal 13.66 70.25 2.76 2.08 5.35 19.56

Peruvian fish meal 9.3 74.64 7.32 1.02 0.76 16.26

Shrimp head meal 10.08 47.44 3.00 12.04 9.74 27.78Rice bran 10.12 14.22 18.40 7.20 50.17 10.01

c NFE-Nitrogen Free Extract

*Means of 2 replicate samplesaWhole feed pea

bDefatted soybean meal

Ingredients 1 2 3 4 5 6 7

% Soybean replacement 0 20 40 60 80 100Neg.

control% Total protein replacement 0 5 10 15 20 25 -

Feed pea - 8.45 16.90 25.36 33.82 42.30 -

Soybean meal 25.00 20.00 15.00 10.00 5.00 - -

Wheat flour 17.00 13.00 9.00 7.00 5.00 5.00 17.00

Rice bran 8.95 9.5 10.05 8.59 7.13 3.65 22.95

Dextrin - - - - - - 49.00

Common ingredients 49.05 49.05 49.05 49.05 49.05 49.05 11.05

Diet No.

*Peruvian fish meal, 23; Ascetes, 3; squid meal, 2; shrimp head meal, 10; seaweed, 2.5; cod liver oil, 2;soybean oil, 1; soybean lecithin, 1; vitamin mix, 1.99; vitamin C, 0.01; mineral mix,1; dicalciumphosphate,1;carboxymethylcellulose, 0.5; ethoxyquin,0.05.

**Vitamin mix. _-carotene ,3.0M.I.U.kg-1;cholecalceferol, 0.6M.I.U.kg-1; thiamine, 3.60g kg-1; riboflavin, 7.20gkg-1; pyridoxine, 6.60g kg-1; cyanocobalamine, 0.02gm kg-1; _-tocopherol, 16.50 gm kg-1; menadione, 2.40gm kg-1; niacin, 14.40gm kg-1; pantothenic acid, 4.00 gm kg-1; biotin, 0.02 gm kg-1 ; folic acid, 1.20gm kg-1;inositol, 30.00gm kg-1; stay C, 100.00gm kg-1; Mineral mix: P, 12.0%; Ca, 12.0%; Mg, 1.5%; Fe, 0.15%; Zn0.42%; Cu, 0.21%; K, 7.50%; Co, 0.011%; Mn, 0.160%; Se, 0.001%; Mo, 0.0005%; Al ,0.0025%; I, 0.04%.


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Table 3 Composition of reference and test diets for in vivodigestibility experiment in shrimp,P. monodonjuveniles (g/100 g feed)

Table 4 Proximate composition (%) of experimental diets*

Table 5 Amino acid composition (per 100 g sample) of soybean meal, feed pea meal and experimental

dietsa

Feed IngredientReference

Diet

Test Diet

(70:30)

Peruvian fish meal 23.00 16.10

Squid meal 2.00 1.40

Ascetes sp. 3.00 2.10

Shrimp head meal 10.00 7.00

Soybean meal, defatted 25.00 17.50

Wheat flour 17.00 11.90

Rice bran 8.90 5.50

Cod liver oil 2.00 1.40

Soy bean oil 1.00 0.70

Vitamin mix 1.99 1.39

Phosphated ascorbic acid* 0.01 0.01

Mineral mix 1.00 0.70

Dicalcium phosphate 1.00 1.00

Seaweed 2.50 1.75

Celufil 0.50 0.50

Erthoxyquin 0.05 0.05

Cr2O31.00 1.00

Feed peas - 30.00

* Phosphitan C, feed grade, Showa Denko K.K. Japan

Diet No. MoistureCrude

ProteinCrude Fat

CrudeFiber

NitrogenFree

ExtractAsh

1 (0) 8.05 40.02 7.34 5.26 30.73 16.65

2 (20) 6.44 39.12 7.06 5.80 31.42 16.60

3 (40) 6.76 39.45 7.69 5.16 31.41 16.29

4 (60) 7.24 39.08 7.06 5.05 33.18 15.63

5 (80) 6.47 39.97 7.40 5.29 31.82 15.52

6 (100) 4.47 39.25 7.71 5.08 32.73 15.23 7 (neg) 7.01 6.98 7.06 5.15 74.89 5.92

*Mean values of 2 replicate samples

0 40 80 100

Aspartic acid 5.32 3.15 2.29 2.28 2.38 2.69

Methionineb

0.63 0.19 0.27 0.29 0.31 0.33

Threonineb

1.72 1.06 0.84 0.86 0.83 0.86

Serine 2.46 1.33 0.91 0.94 0.95 0.97Glutamic acid 8.64 4.67 4.06 3.97 3.77 3.71

Proline 2.11 1.1 1.16 1.21 1.26 1.32

Glycine 2.36 1.17 1.29 1.32 1.34 1.32

Alanine 2.39 1.15 1.24 1.26 1.28 1.36

Valineb

1.41 1.26 1.02 1.04 1.05 1.08

Isoleucineb

1.42 1.22 1.22 1.23 1.23 1.71

Leucineb

4.07 2.2 1.93 1.95 1.96 1.97

Tyrosineb

1.84 0.9 0.87 0.88 0.89 0.89

Phenylalanineb

1.93 1.32 0.96 0.98 1.09 1.05

Lysineb

2.73 2 1.84 1.86 1.84 1.89

Histidineb

1.05 0.57 0.54 0.54 0.53 0.55

Arginineb

2.56 1.9 1.45 1.57 1.69 1.68

aValues are mean of two replicate samples

bEssential amino acids for shrimp, Penaeus monodon (Kanazawa and

Teshima, 1981; NRC, 1993; FDS Manual 1994; Millamena et al., 1996-1999)

Amino acidSoybean

mealFeed pea

meal

% Replacement in the diet


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Table 6 Response of juvenile P. monodonover a 90-d feeding trial to experimental diets containingreplacement levels of soybean meal protein with feed pea meal protein*

Table 7 Apparent digestibility coefficients for dry matter (ADMD) and protein (APD) of feed pea in Penaeusmonodon(%).

Table 8 Apparent digestibility coefficients for dry matter (ADMD) and protein (APD) of formulated diets

containing feed pea in Penaeus monodon(%)

Table 9 Proximate composition (%) of shrimp juveniles after 90 days of culture*

Diet No.

MoistureCrude

Protein(N x 6.25)

Crude FatCrudeFiber

NitrogenFree

Extracta

Ash

Shrimp 1 (0) 11.64 68.26 3.28 2.28 3.69 22.49

Shrimp 2 (20) 7.96 69.98 3.08 2.68 1.9 22.36

Shrimp 3 (40) 9.38 70.10 3.80 2.59 3.61 19.90

Shrimp 4 (60) 7.42 71.54 3.14 3.06 2.56 19.70

Shrimp 5 (80) 6.68 69.12 3.23 2.57 3.6 21.48

Shrimp 6 (100) 11.9 71.21 3.54 2.23 3.67 19.35

Shrimp 7 (neg) 6.35 61.72 2.95 2.58 9.25 23.50*Values are mean of two replicate samplesaNFE-Nitrogen Free Extract

Dietary

treatments% ADMD %APD

Diet 1 (0) 73.40b

82.76c

Diet 2 (20) 70.58b

84.40bc

Diet 3 (60) 71.82b

85.69ab

Diet 4 (80) 77.33a

87.52a

Nutrient digestibility (%)=100-

(100x%Cr2O3diet/%Cr2O3feces x

%nutrient feces/%nutrient diet)

Diet No.Initial mean

wt (g)Final mean

wt (g)

Weight

gaina

(%)SGR

bFCR

cPER

d Survivale

%

1 (0) 0.020.01 1.300.08 5598309a

4.490.06a 1.230.14a 1.230.11a 83

2 (20) 0.020.01 1.130.06 516292a

4.400.01a 1.770.04a 1.440.03a 100

3 (40) 0.020.01 1.450.04 5980165a

4.610.01a 1.360.12a 1.430.01a 75

4 (60) 0.020.01 1.370.01 583965a

4.540.12a 1.890.03a 1.350.02a 92

5 (80) 0.020.01 1.210.18 5187395a

4.380.15a 1.390.32a 1.340.08a 100

6 (100) 0.020.01 1.190.07 5223205a

4.410.04a 1.390.33a 1.660.08a 100

7 (neg) 0.020.01 0.100.02 36482b

1.670.20b 7.021.86b 0.110.01b 75

dPER = weight gain (g) protein intake

eSurvival (%) = Final no. of shrimp/Initial no. of shrimp x 100

*Means of 5 replicate samples. Values in the same row with different superscripts are not significantlydifferent P


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Table 10 Percent recovery of experimental diets in seawater*

AcknowledgementThe authors acknowledge the United States Department of Agriculture and the USA Dry Pea and LentilCouncil for the research and travel funding support; M. Mallare and J. Vera Cruz for the technicalassistance; F. Jarder for the proximate analysis; J. Bangcaya and M. Arnaiz for the amino acid analysis,

and M.J. Bernas for the Cr2O3analysis.

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Duncan, D.B., 1955. Multiple-range and multiple F-tests. Biometrics, 11, 1-42.

Eusebio, P.S., 1991. Effect of dehulling on the nutritive value of some leguminous seeds as protein for tiger prawn, Penaeusmonodon juveniles.Aquaculture 99, 297-308.

Feed Development Section, 1994. Feeds and feeding of Milkfish, Nile Tilapia, Asian sea bass, and Tiger Shrimp. SEAFDECAquaculture Extension Manual No. 21.SEAFDEC Aquaculture Department, Tigbauan, Iloilo, Philippines. 97 pp.

Gomes, L.O.A.and Honculada Primavera, J. 1993. Reproductive quality of male Penaeus monodon . Aquaculture. 112:157-164

Gomes, E.F., Rema, P., Kaushik, S.J., 1993. Effects of dietary incorporation of co-extruded plant protein (rapeseed and peas) ongrowth, nutrient utilization and muscle fatty acid composition of rainbow trout (Oncorhynchis mykiss). Aquaculture 113, 339-353.

Gomes, E.F. Rema, P. Kaushik, S., 1995. Replacement of fish meal by plant protein in the diet of rainbow trout (Oncorhynchismykiss). Digestibility and growth performance. Aquaculture 130, 177-186.

Gouveia, A., Davies, S.J., 1998. Preliminary evaluation of pea seed meal in diets for juvenile European sea bass (Dicentrarchuslabrax). Aquaculture 166, 311-320.

Gouveia, A., Davies, S.J., 2000. Inclusion of an extruded dehulled pea seed meal in diets for juvenile European sea bass(Dicentrarchus labrax). Aquaculture 182, 183-193.

Kanazawa, A., Teshima, S., 1981. Essential amino acids of the prawn. Bulletin of Japanese Society of Scientific Fisheries 17, 1375-1379.

4 8 1 2 1 6 2 4

Diet No.

Diet 1 (0) 89 87 85 83 80

Diet 2 (20) 91 85 83 82 80

Diet 3 (40) 92 91 90 86 85Diet 4 (60) 94 93 93 92 91

Diet 5 (80) 92 91 91 90 90

Diet 6 (100) 91 89 87 88 86

Diet 7 (neg.) 88 83 80 77 76

*Values are means of triplicate samples

No. of hours

% Recovery


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Kaushik, S.J., Vachot, C., Aguirre, P., 1993. Potential utilization of extruded peas. 6th International Symposium on Fish Nutrition,

October 4, 1993. Hobart, Australia.

Millamena, O.M., Teruel, M.B., Kanazawa, A., Teshima, S., 1999. Quantitative dietary requirements of postlarval tiger shrimp,Penaues monodon,for histidine, isoleucine, leucine, phenylalanine and tryptophen. Aquaculture 179, 169-179.

Millamena, O.M., Bautista-Teruel, M.N., Reyes, O.S., Kanazawa, A., 1998. Requirements of juvenile marine shrimp, Penauesmonodon (Fabricius) for lysine and arginine. Aquaculture 164, 95-104.

Millamena, O.M., Bautista, M.N., Reyes, O.S., Kanazawa, A., 1997. Threonine requirement of juvenile marine shrimp Penaeusmonodon. Aquaculture 151, 9-14.

Novoa, N.M.A., Castillo, L.O., 1998. Potentiacialidad del uso de las leguminosas como fuent proteica en alimentos para peces. IV

Simposium Internacional de Nutricion Acuicola. La Paz, B.C.S. Mexico, Noviembre 15-18, 1998.NRC (National Research Council), 1983. Nutreint requirements of warmwater fishes and shellfishes. National Academic Press.

Washington D.C.

Olivera-Novoa, M.A. Olivera-Castillo,L. (Potential use of legumes as protein source in foodstuff of fish)

Santos, J.M., Gomes, E., 1997. Carbohydrates in sea bass (Dicentrarchus labrax) diets: effect of the replacement of fish meal bydifferent sources of carbohydrates on growth, body composition and digestibility. Proc. 3

rd Int. Symp. On Research for

Aquaculture: Fundamental and Applied Aspects, 24-27 August 1997, Barcelona, Spain.186.

Smith, D.M., Tabrett, S.J., Sarac, H.Z. 1999. Fish meal replacement in the diet of prawn, Penaeus monodon. Book of Abstracts of theWorld Aquaculture Society Annual Meeting 99. April 26-May 2, 1999. Sydney, Australia. WAS, Baton Rouge, LA, USA, 707.

Spyridakis, P., Metailler, R., Gabaudan, J., Riaza, A., 1989. Studies on nutrient digestibility in European sea bass (Dicentrarchuslabrax). I. Methodological aspects concerning feces collection. Aquaculture 77, 61-70.

UNIP-ITCF, 1995. In: Carrouee, B., Gatel, F. (eds.) Peas utilization in animal feeding. Union Nationale Interprofessionelle desPlantes Riches en Proteines, Paris, France, 99 pp.


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Feed Pea (Pisum Sativum) As An Alternative Protein SourceIn The Diets Of Nile Tilapia, Oreochromis Niloticus(L.)

Corazon B. Santiago, Perla S. Eusebio, and Timothy P. Welsh

AbstractA study was conducted to test feed pea (Pisum sativum) as a protein source in Nile tilapia diets and todetermine its digestibility. In experiment 1, fishmeal-based diets were used to allow the maximum level of

feed pea as fishmeal protein substitute. Prepared as dry pellets, the test diets were isonitrogenous (30%crude protein) and approximately isocaloric (14 kJ g-1

). Fish meal supplied about 28% protein in the diets.Feed peas (12.7-63.3% of the diet) substituted up to 50% of fishmeal protein. The control diet did notcontain peas. Manually sexed male Nile tilapia, weighing 32-39 g per fish at stocking, were used. The fishwere reared in polyethylene tanks (58x37x27 cm) with aeration. Water was static but was partially changeddaily. Water temperature for the whole duration of the experiment ranged from 22-27C. Fish were allowedto feed to satiation twice daily. The various inclusion levels of feed peas did not affect the body weight,feeding activity and feed efficiency of the tilapia. Weight gain after 9 weeks (range: 21.7+5.6-34.0+9.3 g) didnot differ significantly among treatments (P>0.05). Survival rates were highly variable (40-75%) andmortality was not related to treatment. For experiment 2, plant-based tilapia diets were also designed to beisonitrogenous and isocaloric. Feed peas in the diets ranged from 5.9-41.0%. Feed peas substituted up to35% of the plant protein (or up to 29% of total dietary protein). Separate feeding trials were conducted ontwo strains of all-male tilapia (CLSU and BFAR strains) using the same test diets and larger tanks

(90x78x43 cm). Water temperature during the two trials ranged from 23-27C. After 12 weeks of feeding,weight gain did not differ significantly among treatments in each tilapia strain (45.5+7.8-57.4+12.4 g for theCLSU strain; 52.5+4.4-74.1+9.2 g for the BFAR strain). FCR and PER were not significantly differentamong treatments. Survival was 100% in all tanks in both trials. Determined with the use of chromic oxideas a marker, the apparent protein digestibility of feed pea as ingredient was 87.2+2.0%. Diets inexperiments 1 and 2 that contained peas had more than 90% protein digestibility. Overall, feed pea is analternative protein source that can be used routinely in the diets of Nile tilapia.

Key words: Tilapia nutrition; Peas; Alternative protein source; Fishmeal substitute; Digestibility

IntroductionFish meal is a major source of highly digestible nutrients and is a palatable ingredient in fish diets. It can bethe cheapest protein source, at times, based on the price per kilogram of protein (Hardy, 2000). However,

the demand for fish meal by various food production sectors (e.g., poultry, livestock and aquaculture) isincreasing while the supply is stagnating or even decreasing (Starkey, 1994). This causes the rising cost offish meal. In view of the need to minimize dependence of aquaculture feeds on fish meal, the search foralternative protein sources has been an international concern.

The most commonly used plant protein sources that could partially substitute fish meal in fish diets are oilseed meals, some leaf meals, and cereals and their by-products (Gerpacio and Castillo, 1979; Zamora andBaguio, 1984; Tacon, 1987; NRC, 1993; Hertrampf and Piedad-Pascual, 2000). Various feedstuffs,including grain legumes, have been tested as fishmeal substitutes in farmed tilapia (El-Sayed, 1999). Thegrain legumes are also called pulses, which refer to edible seeds of plants with pods belonging to the familyLeguminocae (Hertrampf and Piedad-Pascual, 2000). Lupins, beans, black and green gram, (feed or field)pea and several other peas (cow pea, chick pea) belong to this category. Although they are used in poultry,swine, and cattle feeds, peas are not among the ingredients ordinarily added in fish diets. However, the

digestibility of several species of legume seeds has been studied in young tilapia (Oreochromis niloticus)(De Silva et al. 1988), juvenile rainbow trout (Oncorhynchus mykiss) (Gomes et al., 1995; Pfeffer et al.,1995; Burel et al., 2000), turbot (Psetta maxima) (Burel et al., 2000), and Australian silver perch (Bidyanusbidyanus) (Allan et al., 2000). Feed pea (Pisum sativum), in particular, has been tested to replace fish mealin the diets of rainbow trout (Gouveia et al., 1993), European sea bass (Dicentrarchus labrax) (Gouveia andDavies, 1998), and Atlantic salmon (Salmo salar) (Carter and Hauler, 2000). Because of its high potentialas a fish feed ingredient, feed pea has to be tested in other important food fish, including tilapia. Thus, thepresent study was conducted to determine the effect of levels of dietary feed peas on the growth andsurvival of juvenile Nile tilapia and on feed efficiency. The digestibility of feed peas as an ingredient and ofdiets containing feed peas was also determined.

Materials and Methods1. Experimental diets

Two sets of diets in dry pellet form were tested in two separate experiments. For experiment 1, six fishmeal-based diets (Table 1) were designed to contain 30% crude protein and digestible energy of about 14 kJ g

-1.

The computed amino acids in the diets, based on the amino acid contents of the ingredients, met or


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exceeded amino acid requirements of young Nile tilapia (Santiago and Lovell, 1988). A fishmeal-based dietallowed the incorporation of high levels of feed peas in the test diets. The feed peas (12.7-63.3% of thediet) substituted 10-50% of fishmeal protein, or about 9-48% of total dietary protein. The diet without feedpeas served as a control. Because of the high nitrogen-free extract (NFE) in feed peas, the diets had aboutthe same energy content, but the supplemental starch and oil decreased as the feed pea in the dietincreased. Based on the actual analysis of protein, lipid, and NFE, however, the digestible energy of thediet with the highest feed pea (63.3%) was slightly lower (Table 1).

In experiment 2, eight plant-based diets containing only 8% fish meal were used (Table 2). The diets werealso designed to be isonitrogenous and isocaloric as in experiment 1. Dietary feed peas (5.9-41.0%)substituted 5-35% of plant protein or about 4-30% of total dietary protein. Substitution of protein fromsoybean meal, copra (coconut) meal and rice bran by the feed peas was maintained at a ratio of 2:1:1.

The fish meal used in experiments 1 and 2 was obtained in two batches. The commercial fish meal,soybean meal and copra meal were ground and sieved using a No. 45 standard testing sieve before use.The dry feed peas were ground whole, including hulls, into powder form. Analyses showed that the peascontained 22.32% crude protein, 1.17% crude fat, 3.36% ash, 3.25% crude fiber, and 58.32% nitrogen-freeextract. These values are close to those reported for feed peas (Muehlbauer and Tullu, 1997; PulseCanada, 1999a; Racz, 1999).

2. Experimental fish and tanksThree strains of tilapia juveniles were obtained for the study. The young fish were reared in the laboratoryfor 6-8 weeks before use. During this time, the fish were fed dry pellets without feed peas. Each batch offish was sorted three times during acclimatization to ensure uniformity in size at stocking.

The fish in experiment 1 were manually sexed male Nile tilapia juveniles (BFS strain) produced in theStation. Each fish weighed 32-39 g at stocking. Twenty-four polyethylene tanks measuring 58x37x27 cmwere filled with 40 l of water and randomly stocked with five fish each.

In experiment 2, two all-male tilapia strains were used in separate feeding trials. The CLSU strainindividually weighed 24-31 g at stocking; the BFAR strain, 25-37 g each. Five tilapia juveniles (CLSU strain)were stocked in each of 24 polyethylene tanks (90x78x43 cm) filled with 180 l of water. In the other feedingtrial, 16 tanks were randomly stocked with six juveniles (BFAR strain) per tank.

3. Feeding treatments, sampling and water managementEach of the feeding experiments was conducted in a completely randomized design. In experiment 1, therewere six dietary treatments with four replicates each. The treatments represented levels of fishmeal proteinsubstitution by the feed peas. In each of the two feeding trials in experiment 2, there were eight treatmentsor levels of plant protein substitution by feed peas. Each of the dietary treatments had three replicates forthe CLSU strain or two replicates for the BFAR strain.

Feeding was done twice daily at 0900 and 1400h for 9 weeks in experiment 1 and for 12 weeks inexperiment 2. The amount of feed given to the fish at each feeding exceeded satiation level to ensure thatthe feed was not limiting. Excess feed was collected 1-1.5 hours after each feeding, dried in an oven,weighed and used in the calculation of feed consumption by difference. The amount of feed consumed wasback calculated to be equivalent to about 5% of fish biomass.

In all feeding trials, body weight of fish was measured weekly for the first 4 weeks and biweekly in thesucceeding weeks except in experiment 1 when the final measurements were done after 9 weeks. Totallength of fish was measured at stocking and during harvest. Feed conversion ratio (FCR, the amount offeed consumed divided by weight gain of fish), protein efficiency ratio (PER, weight gain divided by weightof protein consumed), and survival rate were determined for each tank.

Deep well water stored in a reservoir was used for the growth trials. Water in the rearing tanks was staticbut was partially changed (1/4-1/3 of the volume) daily. All rearing tanks were provided with aeration.Temperature was determined daily in the morning and afternoon. Dissolved oxygen was monitored everymorning before water was partially changed with the use of YSI DO meter (Model 55/25 FT). The pH wasmeasured with a pH meter (Beckman, Model PHI 50). Total ammonia nitrogen (NH 3-N) was determinedweekly before changing of water in the morning by the Nessler method with the use of a Hach kit

(DREL/2010).


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Water temperature for the whole duration of experiment 1 ranged from 22-27C, being at the lower rangefor most of the days. Dissolved oxygen in the morning before partial water change ranged from 3.5-6.2 mg l

-

1. The total NH3-N ranged from 0.63-2.39 mg l

-1and pH, 7.7-7.9. In experiment 2, water temperature for the

feeding trial with the CLSU strain of tilapia was 23-27C; pH, 7.4-8.1; total NH3-N, 0.21-2.42 mg l-1

. For thefeeding trial using the BFAR strain, water temperature also ranged from 23-27C; pH, 7.6-8.7; and totalNH3-N, 0.28-1.74 mg l

-1. For each sampling time, water quality was similar among treatments.

4. Biochemical analyses

Samples of feed ingredients, experimental diets, and some fish at stocking and during harvest wereobtained for proximate analysis by standard methods (AOAC, 1990). At the end of the each feeding trialand after a 24-hour fasting, fish samples were obtained and dissected. The appearance of the fish and theinternal organs were examined visually for any abnormality. Other samples of fish that received the dietswhose digestibility was determined were subjected to cold shock, weighed and kept frozen until they wereprocessed further. The fish samples were then autoclaved, homogenized, dried in a vacuum oven at 60C,and ground for analysis.

5. Digestibility measurements5.1. Digestibility of feed peas as ingredient

The apparent dry matter digestibility (ADMD) and crude protein digestibility (APD) of feed peas asingredient were measured indirectly using chromic oxide (Cr2O3) as external indicator (Cho et al., 1982;Spyridakis et al., 1989). The reference diet for the measurement of digestibility of feed peas was the sameas the control diet used in experiment 1 (Table 1), except that 1% Cr2O3and 1% carboxymethylcellulose(CMC, binder) replaced some amounts of the filler. The test diet contained 70% reference diet and 30%feed peas, following the method of Cho et al. (1982). It also contained 1% Cr2O3 and CMC.

Each of the diets was fed to tilapia in three replicate tanks stocked with 10 juvenile tilapia (37.96 0.42 g

fish-1

). The tilapia came from the same source as those used in experiment 1. The fish were fed at 5-8% of

fish biomass daily with two feedings a day (0830 and 1430h). Water temperature ranged from 24-26C.

A modified Guelph system (Cho et al., 1982; Eusebio and Coloso, 2000) was used in the digestibility trials.Each conical fiberglass tank (250 L capacity) was provided with filtered, flow-through water. Flow rate of thewater was 780-850 ml min

-1. Water passed through a fecal decantation column into an attached clear

plastic bottle where the fecal materials settled until collected. The tanks and the fecal collection apparatus

were cleaned twice daily before feeding in the morning and 2 hours after feeding in the afternoon.Collection of feces released from 1730-0730 h started after a 5-day initial feeding and lasted for 25consecutive days. Fecal material were carefully rinsed with distilled water and stored in a freezer to retardbacterial decomposition. Samples were pooled and freeze-dried. Dry fecal samples and test diets wereanalyzed for Cr2O3(Carter et al., 1960) and crude protein (AOAC, 1990).

5.2. Digestibility of diets containing feed peas

The digestibility of diets with 0, 10, 30 and 50% replacement of fishmeal protein (Table 1), as well as dietswith 0, 15, 25 and 35% replacement of plant protein by feed peas (Table 2), was determined in tilapia inseparate trials. Celufil (filler), starch, and rice bran were adjusted to accommodate 1% Cr2O3and the binderin the diets. Each feeding period, including the 5-day adjustment with the marked diets, lasted for 30 days.

Fish used in determining the digestibility of the two sets of diets containing feed peas were bigger tilapia

juveniles (CLSU strain, 82.7+9.3 g) that came from the same batch used in the growth trial (experiment 2).The digestibility tank system, and the procedure for collection of feces and processing of samples were thesame as in the previous digestibility trial for peas as an ingredient. The apparent digestibility for dry matterand crude protein in the diets was likewise computed using the formula of Spyridakis et al. (1989).

6. Statistical analysis

Data on body weight (weekly and biweekly), weight gain, total length, survival, FCR, PER, carcass proteinand ash, and digestibility coefficients were analyzed by ANOVA using the General Linear ModelsProcedure by SAS for PC (SAS Institute Inc., 1991). When significant difference among treatments wasdetected at P


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However, weight gain was highest at the 10% fishmeal protein substitution, or 12.7% feed pea in the diet.The FCR values for the duration of the experiment, as well as the PER, did not differ significantly (Table 3).Survival rates of tilapia in replicate tanks within treatment were highly variable; mean survival rates did notdiffer significantly among treatments (P>0.05). Mortality was caused by the aggressive behavior of the maletilapia in the small rearing tanks and was not treatment-related.

In experiment 2, body weight of the CLSU strain of tilapia at weekly or biweekly intervals did not differsignificantly among treatments (P>0.05). The final weight gain and length increment, as well as the FCR

and PER, did not differ significantly (Table 4). Survival rate was 100% in all tanks. Likewise, the BFARstrain of tilapia in experiment 2 did not show significant differences in body weight during each sampling, orweight gain and total length increment over a 12-week period (Table 5). Moreover, the FCR and PER didnot differ significantly among treatments. Survival rate in all tanks was also 100%.

Carcass protein of fish from four dietary treatments in experiment 1 showed no significant differences(P>0.05) (Table 6). However, body ash increased significantly in fish fed diets with 37.5 and 63.3% feedpeas (or 30 and 50% fishmeal protein substitution). The fish fed with 10% fishmeal protein substitution(12% feed peas) had the highest carcass lipid. Fish in all other treatments had similar carcass lipid. Inexperiment 2, carcass protein as well as body ash differed significantly among treatments (Table 6), but notrend could be established in relation to fish growth and digestibility of the diets. However, as the dietaryfeed peas in the plant-based diets increased and the dietary lipid decreased (Table 2), carcass lipiddecreased (Table 6).

There was no visible adverse effect of dietary peas on feeding behavior and the appearance of the fish andthe internal organs. The three tilapia strains used in the feeding trials readily accepted the diets containingfeed peas and consumed almost the same amount of feed in relation to their body weight (about 5% of fishbiomass). Visual examination at the end of each trial revealed that the tilapia were generally lean. Fish liverdid not appear fatty and only few adipose tissues were visible in the body cavity.

The apparent dry matter digestibility (ADMD) of feed peas as ingredient was 69.3+1.8% and proteindigestibility (APD) was 87.2+2.0%. The ADMD coefficients for some fishmeal-based diets used inexperiment 1 significantly increased with increasing level of feed peas (Table 7). The APD for the dietstested were all high, exceeding 90%. Although the absolute differences were small for practical purposes,the APD of the control diet (without pea) was significantly higher than that of diet with 12.7% feed pea, but

did not differ significantly from that of diets with 37.5% or 63.3% feed pea (Table 7). The plant-based dietsin which digestibility coefficients were determined also showed a significant increase in ADMD when feedpea in the diet was 29.3% or more (Table 7). The APD coefficients of the plant-based diets were high(>90%), as in the fishmeal-based diets in experiment 1, and did not differ significantly among treatments.

DiscussionThe three feeding trials consistently demonstrated that feed peas could be incorporated in tilapia diets at awide range of levels to replace fishmeal protein or other plant proteins without affecting fish growth, feedefficiency, and survival. The FCR values obtained for all diets in the feeding trials were higher than theexpected values (about 2) and, consequently, the PER values were fairly low. These could be attributed tothe water temperature during the experiments, being lower than the optimum (28-31C) for the tilapia(Luquet, 1991), and to some losses in the collection of the uneaten feed. Because the indoor culture systemhad static water, the dissolved oxygen before partial water change in the morning sometimes dipped close

to the minimum level (3 mg l-1

) for tilapia (Luquet, 1991). Nevertheless, the tilapia in two of three feedingtrials had 100% survival. The growth response of tilapia to diets with peas in the different feeding trials wasnot affected by the dietary levels of feed pea.

In general, carcass protein, lipid and ash did not indicate any adverse effect of increasing dietary feed pea.Although growth did not differ significantly among treatments, significant differences in protein wasobserved in fish fed plant-based diets, but not in fish fed the fishmeal-based diets. Moreover, carcass lipidof the tilapia (CLSU strain) fed plant-based diets decreased with the increase of feed peas in the diet. In acompanion study, feeding of milkfish (Chanos chanos) with varying dietary feed peas caused significantdifferences in growth but it did not influence carcass composition (I. Borlongan, personal communication2002). Similarly, two levels of peas in the diets (20 and 40%) did not affect carcass protein and ash inEuropean sea bass but lipid was significantly lower in fish given diet with 40% pea (Gouveia and Davies,1988).

Although high amounts of feed peas could be used in tilapia diets, the actual amount that would beincorporated in practical applications will be influenced largely by the cost of the peas in relation to the cost


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of competing protein sources. The digestibility of the peas is also considered in evaluating its potential as aprotein source. In the present study, the digestibility coefficients for feed peas in Nile tilapia are comparableto those reported for other freshwater fishes (Table 8). The highest digestibility coefficients for P. sativumwere obtained in turbot, a marine species. In a companion study, the crude protein digestibility of feed peasin milkfish was also higher (92%) than in tilapia; however, dry matter digestibility was lower (I. Borlongan,personal communication 2002). Allan et al. (2000) determined the digestibility of peas and other ingredientsin silver perch as well as that of amino acids in the ingredients. The protein digestibility of the ingredientswas a reflection of the availability of amino acids in silver perch. A few exceptions were the animal protein

sources and some oilseed meals whose proteins may have been damaged during processing (Allan et al.,2000). Moreover, the apparent energy digestibility of peas as ingredient in some fish species (Table 8)could also reflect carbohydrate digestibility in the fish. This is because peas contain much higher amountsof carbohydrate than lipid (Muehlbauer and Tullu, 1997; Pulse Canada, 1999a; Racz, 1999) and the proteindigestibility coefficients for peas in different fish species (Table 8) are close. Moreover, the apparent energydigestibility coefficients reported for peas as ingredient are near the values for the carbohydrate digestibilityof soybean meal (54%), wheat grain (61%), and uncooked corn starch (55-61%) in blue tilapia (0.aureus)(NRC, 1993).

Information on the use of P. sativumin fish diets is limited. Gomes et al. (1995) used feed pea at very a lowlevel (4% of the diet) together with other plant ingredients (faba bean meal, maize gluten, full-fat soybean,and a co-extruded mixture of peas and rapeseed) to partially substitute fish meal in rainbow trout diet. In astudy by Gouveia et al. (1993), pea seed meal was used at 38.2% of the diet (20% of the dietary protein) ofrainbow trout with or without cooking/expansion (145C, 25 kg per cm2). The thermal treatment slightlyimproved the nutritional value of the diets containing the peas. In another study, there was a significantdecrease in the protein digestibility of the diet when the pea (P. sativum) in the diet of rainbow troutincreased from 2550% (Pfeffer et al., 1995). Furthermore, the digestibility of the diets containing either 25or 50% peas increased significantly when the peas were autoclaved. APD coefficients of rainbow trout dietscontaining 25% feed peas increased from 86-91% when the peas had pre-treatment. For diets containing50% peas, APD increased from 83-86% when the peas were autoclaved (Pfeffer et al., 1995). In asubsequent study by Gouveia et al. (1998) on European sea bass (D . labrax), fishmeal-based dietscontaining 20 and 40% pea had APD coefficients of 88-89%. In the present study, the APD coefficients ofthe tilapia diets containing feed pea were high (90.5-92%) and were practically not affected by the dietarylevel of peas. This suggests that feed pea and the ingredients it replaced had similar protein digestibility.The whole peas with hulls were simply dried and ground. The digestibility of dehulled peas would mostlikely be higher in tilapia since dehulling removes most of the fiber of ingredients (Eusebio, 1991). Whencompared to other feedstuffs tested in tilapia, feed pea has APD coefficient that is comparable to that of fishmeal (Hanley, 1987; NRC, 1993; Jauncey, 1998). However, it is slightly lower than the protein digestibilityvalues reported for soybean meal (91-94%) in tilapia species (Hanley, 1987; Lim, 1987; Luquet 1991; NRC,1993).

The protein content of feed peas is close to that of copra (coconut) meal, some cereal grain by-products,brewers' grains, and leaf meals (Gerpacio and Castillo, 1979; Hanley, 1987; Tacon, 1987; NRC, 1993;Hertrampf and Piedad-Pascual, 2000). The relatively low protein content of feed peas compared to fishmeal would limit the amount that could be incorporated in the diets of fish that require high protein levels.However, feed peas could be used as one of the ingredients that would collectively replace fish meal in apractical diet. Feed peas could also replace other plant protein sources, as shown in the present study,especially when its cost is competitive. Feed peas contain high levels of lysine (Pulse Canada, 1999a;

Racz, 1999) which could complement the amino acids of other fishmeal substitutes that are deficient inlysine (e.g. canola meal and gluten meals). However, as in soybean meal and other legume seeds, thesulfur amino acids (methionine and cystine) are low and could be the first limiting amino acids when highamounts of plant protein sources are used. For fish requiring high dietary protein, pea protein concentrate(49% crude protein) has been explored as a possible partial replacement of fish meal (Carter and Hauler,2000). Up to 27% pea protein concentrate in the extruded feed did not affect growth of salmon parr. Cowpea (Vigna unguiculata) protein concentrate was also tested in Nile tilapia fry and up to 40% inclusion levelin the diet gave acceptable results (Olvera-Novoa et al., 1997).

One important concern in the use of pulses as ingredients in aqua feeds is the presence of anti-nutritionalfactors (Tacon, 1987; Hertrampf and Piedad-Pascual, 2000). However, through genetic selection, presentpea varieties contain no tannins and only low levels of trypsin inhibitors and lectins (Bond and Duc, 1993)that can be deactivated by heat treatment (Melcion and van del Poel, 1993). Pulses are produced mainly for

their mature seeds and immature pods for human consumption (Muehlbauer and Tullu, 1997), unlikesoybean and other oilseeds that are grown mainly for processing into edible oils and protein concentrates.With the expansion of production of feed peas as human food and animal feed (Pulse Canada, 1999b),more peas would be available in the market.


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The consistent response of the tilapia juveniles to diets containing feed peas in three feeding trials clearlyshowed that feed pea as an alternative ingredient can be routinely included in tilapia diets. Further studieson feed peas as component of tilapia diets (e.g., those related to processing and energy utilization) have tobe conducted.

Table 1 Composition of fishmeal-based diets for male Nile tilapia in experiment 1 (g per 100g diet)

Table 2 Composition of plant-based diets for male tilapia (CLSU and BFAR strains) in experiment 2(g per 100g diet)

Table 3 Mean initial weight and total length, gain in weight and length, survival, FCR and PER of male Niletilapia fed diets containing feed peas at varying fishmeal protein substitution for 9 weeks

(experiment 1)a

Ingredient 0a

5 (4.2) 10 (8.4) 15 (12.7)a

20 (16.9) 25 (21.1)a

30 (25.3) 35 (29.6)a

Fish meal 8.00 8.00 8.00 8.00 8.00 8.00 8.00 8.00

Soybean meal 40.00 38.63 37.25 35.88 34.50 33.13 31.75 30.38

Copra meal 20.00 18.37 16.73 15.10 13.47 11.83 10.20 8.57

Rice bran 24.00 21.49 18.98 16.46 13.95 11.44 8.93 6.41

Peas - 5.86 11.72 17.57 23.43 29.29 35.15 41.00

Soybean oil 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00

Vitamin mix 0.40 0.40 0.40 0.40 0.40 0.40 0.40 0.40

Mineral mix 0.70 0.70 0.70 0.70 0.70 0.70 0.70 0.70

Starch 2.90 2.56 2.23 1.89 1.55 1.21 0.88 0.54

Analyzed (as fed)

Crude protein (%) 29.7 32.4 32.4 31 32 29.8 31.8 29.9

Crude fat (%) 10.1 9.5 8.7 7.1 8.2 7.5 7.1 6.9

Ash (%) 7.7 6.3 7 5.6 7.2 7 6.2 6.6

Crude fiber (%) 7.9 7.4 7.7 6.5 7.2 6.6 6.6 6.3

NFE (%) 37.4 37.2 37.4 42.9 38.3 41.8 41.2 43

Digestible energy (kJ g-1

) 14.2 14.4 14.2 14.1 14.1 13.9 14 13.9aDiet whose digestibility in tilapia was also determined in a separate feeding trial.

% replacement of plant proteins (or total dietary protein)

0 a 10 (9.3)a 20 (18.6) 30 (28.0) 40 (37.3) 50 (47.6)Fish meal 49.0 44.1 39.2 34.3 29.4 24.5

Peas - 12.7 25.3 37.5 50.7 63.3

Rice bran 15.0 15.0 15.0 15.0 15.0 10.2

Starch 15.0 12.0 9.0 6.0 - -

Soybean oil 5.0 5.0 5.0 5.0 2.9 -

Vit-min mix 2.0 2.0 2.0 2.0 2.0 2.0

Celufil (filler) 14.0 9.2 4.5 0.2 - -

Analyzed (as fed)

Crude protein (%) 31.9 31.6 31.0 29.9 29.2 28.9

Crude fat (%) 11.2 11.0 10.4 10.4 7.7 2.5

Ash (%) 9.7 9.0 8.9 8.1 7.7 6.7

Crude fiber (%) 6.7 6.5 6.2 6.8 7.0 6.4

NFE (%) 33.9 35.7 37.2 37.8 41.3 48.6

Digestible energy (kJ g-1) 14.5 14.7 14.6 14.4 13.8 12.9

% replacement of fishmeal protein (or total dietary protein)

aDiet whose digestibility in tilapia was also determined in a separate feeding trial.

Ingredient

Initial Gain Initial Gain

- - 35.5+0.9 24.0+7.7 12.6+0.7 3.0+0.5 40+23 4.1+0.7 0.8+0.1

10 12.7 35.3+0.6 34.0+9.3 12.5+0.6 3.4+0.4 70+26 3.2+0.5 1.0+0.2

20 25.3 35.8+0.3 25.3+7.3 12.6+0.5 2.8+0.4 70+26 4.2+1.6 0.8+0.3

30 37.5 34.9+1.1 21.7+5.6 12.5+0.6 2.4+0.1 55+30 4.2+0.6 0.8+0.140 50.7 35.5+0.6 25.1+4.6 12.7+0.6 3.0+0.6 45+19 3.4+0.7 1.0+0.2

50 63.3 35.5+0.6 25.2+9.6 12.4+0.7 2.8+0.7 65+25 4.0+1.4 1.0+0.4

Total length (cm)

aColumn means are not significantly different from each other (P>0.05). Mean+SEM

Fishmeal proteinsubstitution (%)

Dietary feedpeas (%)

FCRSurvival

(%)PER

Body weight (g)


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Table 4 Mean initial weight and total length, gain in weight and length, FCR and PER of tilapia (CLSUstrain) fed diets containing feed peas at varying plant protein substitution for 12 weeks(experiment 2)

a

Table 5 Mean initial weight and total length, gain in weight and length, FCR and PER of male tilapia (BFAR

strain) fed diets containing feed peas at varying plant protein substitution for 12 weeks

(experiment 2)a

Table 6 Carcass protein and ash of Nile tilapia after 9 weeks of feeding using some diets in experiment 1

and after 12 weeks of feeding in experiment 2a


0 0 28.0+3.0 53.6+12.8 11.6+0.1 4.7+1.4 3.4+0.6 1.0+0.2

5 5.9 28.2+2.8 57.4+12.4 11.6+0.3 5.0+0.2 3.2+0.4 1.0+0.1

10 11.7 28.5+3.2 49.5+ 7.2 11.5+0.2 5.1+0.6 3.6+0.4 0.9+0.115 17.6 28.3+2.9 57.1+ 8.5 11.7+0.2 5.2+0.9 3.3+0.3 1.0+0.1

20 23.4 28.5+2.6 56.7+ 2.1 11.7+0.2 5.1+0.3 3.3+0.3 1.0+0.1

25 29.3 28.4+3.2 45.5+ 7.8 11.7+0.1 4.6+0.4 4.0+0.4 0.9+0.1

30 35.2 28.4+3.0 54.0+ 0.7 11.6+0.2 5.1+0.2 3.5+0.2 0.9+0.0

35 41 28.1+2.4 56.7+13.5 11.5+0.1 5.4+1.0 3.4+0.6 1.0+0.2


Total length (cm)Dietary feedpeas (%)

Plant proteinsubstitution (%) FCR PER

Body weight (g)


0 0 31.1+4.3 52.5+4.4 12.1+0.6 5.0+0.6 4.3+0.4 0.8+0.1

5 5.9 31.0+3.6 71.6+22 12.1+0.5 5.8+0.8 3.3+0.6 0.9+0.2

10 11.7 30.9+4.3 60.2+0.7 12.1+0.6 5.0+0.4 3.6+0.2 0.8+0.0

15 17.6 30.9+4.3 69.0+11.5 12.1+0.5 4.8+1.5 3.7+0.6 0.9+0.2

20 23.4 30.8+3.9 62.4+10 12.2+0.6 5.1+0.3 3.6+0.5 0.9+0.1

25 29.3 30.9+3.6 57.0+5.9 12.1+0.5 5.2+0.8 3.6+0.1 0.9+0.0

30 35.2 31.0+3.8 63.9+1.3 12.1+0.4 5.1+0.3 3.8+0.1 0.8+0.0

35 41 31.1+4.5 74.1+9.2 12.1+0.6 5.7+0.7 3.2+0.1 1.0+0.0

FCR PER


Total length (cm)Plant proteinsubstitution 0.80


Body weight (g)

Crudeprotein

Ash Lipid

Diets in Expt 1

0 - 56.5 0.2a

18.9 0.1b

18.80.3b

10 12.7 56.4 0.6a

19.5 0.3b

19.40.0a

30 37.5 57.2+0.4a

20.8+0.4 a

18.70.0b

50 63.3 57.4+0.2a

20.8+0.1 a

18.90.1b

Diets in Expt 2b

0 - 55.7+0.4c

16.3+0.6 b

23.20.4a

15 17.6 59.1+0.4a

14.7+0.2c

21.20.3b

25 29.3 56.5+0.2c

16.3+0.3 b

20.90.3b

35 41.0 58.2+0.2b

18.3+0.0 a

20.30.1b

aFor each experiment, column means with a common superscript are not

significantly different (P>0.05).

bAt stocking of the CLSU tilapia strain, body protein was 58.7+0.5%; ash,

26.0+0%; lipid, 11.3+0.2%.

Dry matter basis (%)Diet


% fishmeal protein substitution

% plant protein substitution


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Table 7 Apparent dry matter digestibility (ADMD) and protein digestibility (APD) of diets with peas injuvenile male tilapia (CLSU strain)

Table 8 Digestibility coefficients (%) for P. sativumin different fish species

AcknowledgementsThis study was funded by the US Department of Agriculture and sponsored by the USA Dry Pea and LentilCouncil. Feeds and fish samples were analyzed by the staff of the Central Analytical Laboratory ofSEAFDEC AQD, Tigbauan, Iloilo or the Animal Nutrition Analytical Service Laboratory, University of thePhilippines Los Baos, Laguna.

ReferencesAllan, G.L., Parkinson, S., Booth, M.A., Stone, D.A.J, Rowland S.J., Frances, J., Warner-Smith, R., 2000. Replacement of fish meal in

diets for Australian silver perch, Bidyanus bidyanus: 1. Digestibility of alternative ingredients. Aquaculture 186, 293-310.Association of Official Analytical Chemists (AOAC), 1990. Official Methods of Analysis, 15th edn. AOAC, Arlington, VA, 1228 pp.Bond, D.A., Duc, G., 1993. Plant breeding as a means of reducing antinutritional factors in grain legumes. In: van der Poel, A.F.B.,

Huisman, J., Saini, H.S. (Eds.), Second International Workshop on Antinutritional Factors (ANFs) in Legume Seeds, RecentAdvances of Research in Antinutritional Factors in Legume Seeds, 1-2 December 1993, EAAP Publication, The Netherlands, pp.379-396.

Burel C., Boujard, T., Tulli, F., Kaushik, S.J., 2000. Digestibility of extruded peas, extruded lupin, and rapeseed meal in rainbow trout(Oncorhynchus mykiss) and turbot (Psetta maxima). Aquaculture 188, 285-298.

Carter, J.F., Bolin, D.W., Erikson, P., 1960. The evaluation of forages by the Agronomic difference method and chromogen chromicoxide indicator technique. N.D. Agric. Exp. Stn., Techn. Bull. 426, 55 pp.

Carter, C.G., Hauler, R.C., 2000. Fish meal replacement by plant meals in extruded feeds for Atlantic salmon, Salmo salar L.

Aquaculture 185, 299-311.Cho, C.Y., Slinger, S.J., Bayley, M.S., 1982. Bioenergetics of salmonid fishes: energy intake, expenditure and productivity. Comp.

Biochem. Physiol. 73B, 25-41.De Silva, S.S., Keembiyahetty, C.N., Gunasekera, R.M., 1988. Plant ingredient substitutes in Oreochromis niloticus (L.) diets:

Ingredient digestibility and effect of dietary protein content on digestibility. J. Aqua. Trop. 3, 127-138.El-Sayed, A.-F. M., 1999. Alternative dietary protein sources for farmed tilapia, Oreochromis spp. Aquaculture 179, 149-168.Eusebio, P., 1991. Effect of dehulling on the nutritive value of some leguminous seeds as protein sources for tiger prawn (Penaeus

monodon) juveniles. Aquaculture 99, 297-308.Eusebio, P.S., Coloso, R.M., 2000. Nutritional evaluation of various plant protein sources in diets for Asian sea bass Lates calcarifer.

J. Appl. Ichthyol. 16, 56-60.Gerpacio, A.L., Castillo, L.S., 1979. Nutrient Composition of Some Philippine Feedstuffs. Technical Bulletin 21, Extension Division,

Department of Animal Science, University of the Philippines at Los Baos, Philippines, 117 pp.Gomes, E.F., Rema, P., Kaushik, S.J., 1995. Replacement of fish meal by plant proteins in the diet of rainbow trout (Oncorhynchus

mykiss): digestibility and growth performance. Aquaculture 130, 177-186.Gouveia, A., Oliva Teles, A., Gones, E., Rema, P., 1993. Effect of cooking/expansion of three legume seeds on growth and food

utilization by rainbow trout. In: Fish Nutrition in Practice, Biarritz (France), June 24-27, 1991. Ed. INRA, Paris (Les Colloques,

no61), pp. 933-938.Gouveia A., Davies, S.J., 1998. Preliminary nutritional evaluation of pea seed meal (Pisum sativum) for juvenile European sea bass

(Dicentrarchus labrax). Aquaculture 166, 311-320.

Digestibilitytrials


ADMD (%)a

APD (%)a

0 0 72.1 0.6c 92.5 0.4a

10 12.7 75.6 0.4b

90.5 0.4b

30 37.5 77.8+1.1ab

91.5+0.7ab

50 63.3 78.8+0.6a

92.0+0.2ab

0 0 74.1+0.2b

90.6+0.3a

15 17.6 73.4+0.59b

91.5+0.3a

25 29.3 76.2+0.67a

91.2+0.3a

35 41 76.9+0.42a

91.6+0.3a

Trial using diets in Expt 1

Trial using diets in Expt 2

aFor each digestibility trial, column means with a common

superscript are not significantly different (P>0.05).

% plant protein substitution

% fishmeal protein substitution

Fish Dry matter Protein Energy Reference

Rainbow trout, 52 g 66.1 80.4 59.2 Gomes et al. (1995)

Rainbow trout, 100g 66.3 87.9 68.9 Burel et al. (2000)

Turbot, 110 g 71.5 92.9 77.7 Burel et al. (2000)

Silver perch 51.0 81.0 51.0 Allan et al. (2000)

Nile tilapia, 38 g 69.3 87.2 no data present study


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Hanley, F., 1987. The digestibility of feedstuffs and the effects of feeding selectivity on digestibility determinations in tilapia,Oreochromis niloticus. Aquaculture 66, 163-179.

Hardy, R.W., 2000. Fish feeds and nutrition- Fish feeds & nutrition in the new millenium. Aquaculture Magazine, Jan/Feb 2000, pp.85-89.

Hertrampf, J.W., Piedad-Pacual, F., 2000. Handbook on Ingredients for Aquaculture Feeds. Kluwer Academic Publishers, TheNetherlands, 573 pp.

Jauncey, K., 1998. Tilapia Feeds and Feeding. Pisces Press Ltd., Stirling, Scotland, 241 pp.Lim, C., 1987. Practical feeding tilapias. In: Lovell, T., Nutrition and Feeding of Fish. Von Nostrand Reinhold, NY, pp 163-183.Luquet, P., 1991. Tilapia, Oreochromis spp. In: Wilson, R.P. (Ed.), Handbook of Nutrient Requirements of Finfish. CRC Press, Inc.

Florida, USA, pp. 169-179.

Muehlbauer, F.J. and Tullu, A., 1997. Pisum sativum L. http://www.hort.purdue.edu/cropfactsheets/pea.htm (2 July 2002).Melcion, J.-P., van der poel, A.F.B., 1993. Process technology and antinutritional factors: principles, adequacy and process

optimization. In: van der Poel, A.F.B., Huisman, J., Saini, H.S. (Eds.), Second International Workshop on Antinutritional Factors(ANFs) in Legume Seeds, Recent Advances of Research in Antinutritional Factors in Legume Seeds, 1-2 December 1993, EAAPPublication, Wageningen, The Netherlands, pp. 419-434.

National Research Council (NRC), 1993. Nutrient Requirements of Fish. National Academy Press, Washington, DC., 114 pp.Olvera-Novoa, M.A., Pereira-Pacheco, F., Olivera-Castillo, L., Prez-Flores, V., Navarro, L., Smano, J.C., 1997. Cowpea (Vigna

unguiculata) protein concentrate as replacement of fish meal in diets for tilapia (Oreochromis niloticus) fry. Aquaculture 158, 107-116.

Pfeffer, E., Kinzinger, S., Rodehutscord, M., 1995. Influence of the proportion of poultry by-products and of untreated orhydrothermically treated legume seeds in diets for rainbow trout, Oncorhynchus mykiss (Walbaum), on apparent digestibilities oftheir energy and organic compounds. Aquaculture Nutr. 1, 111-117.

Pulse Canada, 1999a. Nutrient Composition. http://www.pulsecanada.com/FeedPeas/NutrientComposition.htm (29 June 2002).Pulse Canada, 1999b. Production Info. http://www.pulsecanada.com/FeedPeas/ProductionInfo.htm (29 June 2002)Racz, V.J., 1999. Feed pea nutrient composition. In: Feed Peas, Selected Publications and Articles for Animal Nutritionists and

Livestock Producers. USA Dry Pea and Lentil Council, Bangkok, Thailand, pp. 5-8.

Santiago, C.B., Lovell, R.T., 1988. Amino acid requirements for growth of Nile tilapia. J. Nutr. 118, 1540-1546.SAS Institute Inc., 1991. SAS System for Linear Models. Third Edition. SAS Institute Inc., Cary, North Carolina, 329 pp.Spyridakis, P., Metailler, R., Gabaudan, J., Riaza, A., 1989. Studies on nutrient digestibility in European sea bass (Dicentrarchus

labrax). I. Methodological aspects concerning feces collection. Aquaculture 77, 61-70.Starkey, T. J., 1994. Status of fish meal supplies and market demand. Miscellaneous report. H.J. Baker and Bro. Inc., Stamford, CT.,

USA, 28 pp.Tacon, A.G.J., 1987. The Nutrition and Feeding of Farmed fish and Shrimp - A Training Manual 2. Nutrient Sources and Composition.

FAO, Brazil, 129 pp.Zamora, R.G., Baguio, S.S., 1984. Feed Composition Tables for the Philippines. PCARRD Book Series No. 1/1984, Philippine Council

for Agriculture and Resources Research and Development, Los Baos, Laguna, Philippines, 264 pp.


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Potential Of Feed Pea (Pisum Sativum) As An Alternative Protein SourceIn Practical Diets For Milkfish (Chanos ChanosForsskal)

Ilda G. Borlongan, Perla S. Eusebio, and Timothy P. Welsh

AbstractA 12-week feeding trial was conducted to evaluate the use of feed pea as a potential dietary protein sourcefor juvenile milkfish, Chanos chanosForsskal. Six isonitrogenous (30% crude protein) and isocaloric (3940

kcal/kg) practical diets were formulated. The control diet contained fish meal, soybean meal, meat andbone meal, and copra meal as principal protein sources. Feed pea was progressively substituted at 0, 5,10, 15, 20, 25 and 30% of total protein. A leading commercial milkfish feed was also tested as an additionalcontrol. The experimental diets were fed to triplicate groups of milkfish fingerlings (mean initial weight of0.42 0.01g) at 10% body weight/day. Growth performance (expressed as % weight gain and SGR),survival, feed conversion ratio (FCR) and protein efficiency ratio (PER) of milkfish fed diets with up to 10%substitution of the dietary protein with feed pea were not significantly different (P >0.05) compared to fishfed the control diet. Replacement with feed pea at 15% and higher levels led to milkfish fed these dietsshowing a significantly lower growth response compared to fish fed the control without any feed pea.Nevertheless, it was observed that milkfish fed diets with up to 20% of total dietary protein substitution withfeed pea showed better growth rates and feed conversion ratios than the commercial feed control. Wholebody composition (crude protein, crude fat, crude fiber, nitrogen free extracts, and ash content) of milkfishfed the various test diets was not significantly different. Apparent digestibility coefficients of feed pea and

experimental diets in milkfish were also determined. Results indicate that feed pea is an acceptable proteinsource and can substantially replace up to 20% of the total dietary protein in milkfish diets.

Keywords: Aquaculture feeds; alternative protein source; feed pea (Pisum sativum);milkfish (Chanos chanos Forsskal)

IntroductionThe current trend in milkfish culture is toward increased intensification of culture systems whereby provisionof feeds becomes necessary and success therefore depends significantly on the availability of well-balanced, nutritionally complete and cost-effective feeds. For many years, SEAFDEC AQD has doneconsiderable research on the nutrient requirement of milkfish, assessment of nutritive value of availableingredients, and development of simple and appropriate feeding technology. These are all importantfactors toward the development of cost-effective feeds and feeding strategy. SEAFDEC AQD has also

endeavored to develop cost-effective feeds for milkfish culture. There is a need, however, for these feedsto be continuously refined, improved, and tested for technical and economic feasibility.

In recent years, the rising cost, uncertain availability, and fluctuating quality of fish meal has led to thesearch for alternative protein sources for fish feed to sustain fish production. The use of plant proteinsources to completely or partially replace fish meal in fish diets has been studied for many years by severalworkers for different fish species (De la Higuera et al., 1988; Moyano et al.,1992; Borlongan and Coloso,1994; Sanz et al., 1994; Gomes et al., 1995; Kaushik et al., 1995; Hardy, 1996; Robaina etal., 1997;Watanabe etal., 1997; Carter and Hauler, 2000; Kissil et al., 2000 and Farhangi and Carter, 2001).

Feed pea (Pisum sativum), an abundant agricultural product, is a potential feed ingredient. It is a highenergy, medium protein ingredient with average protein content (%CP) of 22 to 24% and digestible energy(DE) of 3420 kcal/kg. Lysine is particularly high at 1.6%. The efficiency of feed pea as a feed ingredient

has been evaluated for cattle, swine, and poultry. However, there is a paucity of information on the use ofthis ingredient as dietary protein source for aquaculture species.

The aim of the current study was to assess the potential of feed pea ( Pisumsativum) as an ingredient inpractical feeds for milkfish (Chanos chanosForsskal). This was compared with solvent-extracted soybeanmeal because soybean meal is the most often used plant alternative to fish meal in milkfish feeds. The firstphase of the experiment was conducted to establish the nutritive value of feed pea through measurement offish growth, feed utilization and carcass composition. The second phase assessed the apparentdigestibility coefficients for dry matter and dietary protein.

Materials and MethodsExperiment 1: Growth performance and feed utilization

Experimental DietsThe test diets were formulated to be isonitrogenous and isocaloric. The control diets were the existingSEAFDEC milkfish formulation that had been previously tested in intensive ponds as having an FCR of 1.5(D-1), and a leading commercial milkfish feed (D-8). The composition and proximate analyses of the


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experimental diets are presented in Table 1. All diets were formulated to contain 30% protein, 10% lipidand energy of about 394kcal/100g diet. Analyses however, showed that experimental diets contained 33%protein and 10% lipid, while the commercial milkfish feed control contained 37% protein and 9% lipid. Thebasal formulation contained fish meal, defatted soybean meal, meat and bone meal, and copra meal asdietary protein sources. The levels of feed pea incorporation tested were 0, 5, 10, 15, 20, 25, and 30% ofthe total crude protein. Dietary essential amino acid profiles (Table 2) were calculated based on analysesof ingredients and published data (NRC, 1977) and compared to that of requirements for milkfish juvenile(Borlongan and Coloso, 1993) which was used as the reference. Analyzed values of the essential amino

acids are likewise presented. No supplemental amino acids were added to the diets. Cod liver andsoybean oils served as lipid sources. Vitamin and mineral premixes were kept constant in all diets.

Commercial feed peas of US origin were sourced from a local agricultural distributor. The whole dry peawas oven-dried at 60

0C for 4 hours and finely ground to homogenized flour. No specialized ingredient

processing method was used. Diets were prepared by first mixing all dry ingredients in the Hobart mixer.Oils were then blended with the dry ingredient mixture. An equal portion of bread flour was gelatinized bycooking in 600 ml water and added to the mixture. The semi-moist mixture was then passed through theHobart food grinder to form 2 mm diameter pellets. The pellets were dried in an air convection oven at40

0C. The dry pellets were then ground, sieved to uniform sizes, and stored at 4

0C until used for feeding.

Experimental fish and feeding management

The experiment was conducted at the Feed Development Section of the Aquaculture Department of theSoutheast Asian Fisheries Development Center (SEAFDEC/AQD), Tigbauan, Iloilo, Philippines. Hatchery-bred milkfish juveniles were acclimated for two weeks under laboratory conditions and to a dry diet (controldiet) prior to the experiment. The feeding trials were conducted in an indoor flow-through system. Sixty-litercapacity, oval fiberglass tanks containing 50L seawater with aeration were used. Milkfish juveniles (meanwt. = 0.42 g) were randomly distributed at a stocking rate of 10 fish per tank and in four replicate tanks pertreatment. Fish in each treatment were then fed three times daily at 0830, 1130, and 1400H at a feedingrate of 10% body weight per day for 12 weeks. Feeding ration was adjusted at every 3 weeks samplinginterval. Water quality parameters (temperature, salinity, DO, pH, ammonia, nitrite) were monitoredfollowing the methods of Strickland and Parsons (1972) to ensure water quality remained well within limitsrecommended for milkfish culture (Bagarinao, 1991).

Statistical Analyses

Survival, % weight gain, specific growth rate (SGR), food conversion ratio (FCR), and protein efficiencyratio (PER) were calculated and subjected to statistical analyses. Data were subjected to analysis ofvariance (ANOVA) and Duncans Multiple Range test (DMRT). Differences between means wereconsidered significant at P


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1% chromic oxide to replace an equivalent amount from the filler (Celufil). The fish were acclimated withthe control diet/reference diet (without Cr2O3) for 1 week prior to feeding them test diets containing 1%Cr2O3. Diets were fed to satiation twice daily (0900 and 1400 h). Fecal collection was started at day 5 afterthe fish were acclimated to the green diets, or when the fecal matter became greenish. The tanks and fecalcollection apparatus were cleaned twice daily, 2 hours after feeding in the morning and afternoon. Fecalcollection bottles were then attached, and feces collected. Feces were collected from the plastic bottles,rinsed 3 times with distilled water; freeze-dried and prepared (Eusebio, 1991) for dry matter and crudeprotein (AOAC, 1990) and Cr2O3(Carter et al., 1960) analyses. The test diets were likewise analyzed for

dry matter, crude protein and Cr2O3.

In vivoapparent protein digestibility (APD) and apparent dry matter digestibility (ADMD) of feed pea werecomputed using the formula of Spyridakis et al. (1989) and Cho et al. (1982). ADMD and APD offormulated diets were computed using the formula of Spyridakis et al. (1989). The data were analyzedusing ANOVA for completely randomized design (CRD). Treatment means were compared by Duncan'sMultiple Range Test (DMRT) using SAS computer software. Differences were considered significant atP


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The results of the digestibility trial did not support the assumption of reduced protein availability from feedpea. In the present study, dietary crude protein digestibility values of the experimental diets were above80%. A comparison with the control diet supports the observation that pea protein is well digested bymilkfish. In fact, apparent protein digestibility value was superior with the partial inclusion (10% of dietaryprotein) of this plant material compared to the control without feed pea. Feed pea at this level may onlyshow marginal effects of anti-nutritional factors and limiting amino acids due to its relatively low content ofsuch compounds and its fairly good protein biological value for fish nutrition. Another consequence of usingfeed pea in place of soybean meal in the diet formulation for milkfish is the reduction of bread flour in diets

containing high levels of feed pea. This is because of the higher carbohydrate content of feed peacompared to soybean meal. At this inclusion level, in practical conditions, feeding costs can be dramaticallydecreased.

In general, feed pea meal proved to be an acceptable ingredient for consideration in practical diets forgrower stage production of milkfish. Future work showed be directed towards evaluating extruded peaproducts where pre-gelatinization of the starch would be possible, or testing various pea proteinconcentrates which have

Documents

Utilization of Feed Peas (Pisum Sativum)