Final Project Report

Title: Utilization of feed peas (Pisum sativum) as alternative protein sources in diets for
shrimp, tilapia, and milkfish.
Implementing Personnel and Agencies:
Southeast Asian Fisheries Development Center, Aquaculture Department
Clarissa L. Marte, PhD, Head, SEAFDEC Research Division (clmarte@aqd.seafdec.org.ph)
Myrna B. Teruel, Study leader, Shrimp (mbt@aqd.seafdec.org.ph)
Corazon B. Santiago, PhD, Study leader, Tilapia (csantiago@aqd.seafdec.org.ph)
Ilda G. Borlongan, Study leader, Milkfish (igb@aqd.seafdec.org.ph)
Perla S. Eusebio, Study leader, Digestibility experiments (pse@aqd.seafdec.org.ph)
USA Dry Pea and Lentil Council
Timothy Welsh, Southeast Asia Representative (agsource@loxinfo.co.th)
Funding: US Department of Agriculture, Emerging Markets Program
USA Dry Pea and Lentil Council
Trial Period: July, 2001 through March, 2002
Table of Contents
SUMMARY ............................................................................................................................................................ 1-2
STUDY 1. SHRIMP (PENAEUS MONODON, FABRICIUS)............................................................................................... 1
STUDY 2. TILAPIA (OREOCHROMIS NILOTICUS L.)..................................................................................................... 1
STUDY 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 Source
In 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.......................................................................................................................................................... 20
MATERIALS AND METHODS...................................................................................................................................... 20
RESULTS.................................................................................................................................................................... 22
DISCUSSION............................................................................................................................................................... 22
ACKNOWLEDGEMENT............................................................................................................................................... 26
REFERENCES.............................................................................................................................................................. 26
1
SUMMARY
This report summarizes the results of experiments conducted to test the efficacy of feed pea meal (Pisum
sativum) as an alternative protein source in the diets for tilapia, milkfish, and shrimp. All formulations were
based on the known nutrient requirements (e.g., protein, fat, carbohydrate, protein:energy ratio,
indispensable amino acids) of the species specified.
Study 1. Shrimp (Penaeus monodon, Fabricius)
The potential of feed pea meal as an alternative protein source in practical diets for juvenile tiger shrimp
was assessed in a 12-week feeding trial. Formulated diets were made isonitrogenous (40% crude protein)
and isocaloric (14.5 kJ g
1
). The control diet contained fish meal, soybean meal, squid meal, shrimp meal,
shrimp head meal as protein sources. 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, 25%,
respectively, of the total protein in the diet. A negative control with no protein sources was added to the
treatment. Twelve shrimp post-larvae with an average weight of 0.020.01g were randomly assigned in 35,
60-l oval tanks equipped with a flow-through seawater system. The shrimp were fed the formulated diets at
a daily feeding rate of 20-25% body weight in 5 replicate samples.
Results showed no significant differences in % weight gain (WG)(5162 to 5839); specific growth rate (SGR)
(4.4-4.6); feed conversion ratio (FCR) (1.2-1.9); protein efficiency ratio (PER) (1.2-1.7); survival (SURV)
(75-100) of shrimp fed diets with 0 up to the highest level of replacement. Weight gain of shrimp fed the
negative control (364) was significantly lower (P>0.05) compared to the rest of the treatments.
The apparent dry matter (ADMD) and protein (APD) digestibilities of the dry feed pea in P. monodon were
high at 73.384.98% and 92.742.62%, respectively. Digestibility coefficients for dry matter (71-77%) and
protein (83-88%) for the feed pea meal-based diets increased with increasing level of feed pea
replacement.
The results indicate that whole feed pea meal can be an alternative protein source for shrimp. It can
substitute even up to 100% of the protein from defatted soybean meal, which is equivalent to 25% of the
total protein in the diet. An inclusion level of up to 42% in the juvenile shrimp practical diet did not manifest
any adverse effects on growth, feed efficiency, survival, body composition, and digestibility coefficients for
dry matter and protein of the shrimp.
Study 2. Tilapia (Oreochromis niloticus L.)
The tilapia experiments made use of fish meal-based (experiment 1) and plant-based (experiment 2) diets
to allow maximum level of feed pea as a protein source. In both experiments, all test diets were made
isonitrogenous (30% crude protein) and isocaloric (14 kJ g
1
). Water temperature during the feeding trials
ranged from 22-27C
Experiment 1. Fish meal supplied about 28% protein in the diets. Feed peas (12.7-63.3% of the diet)
substituted up to 50% of fish meal protein. Manually sexed male Nile tilapia, each weighing 32-39g at
stocking, were used. The fish were reared in polyethylene tanks (58x37x27 cm) with aeration under
controlled conditions. Water was static and was partially replaced once a day. Fish were allowed to feed to
satiation twice daily.
The various inclusion levels of feed peas did not affect body weight, feeding activity, and feed efficiency of
the tilapia. Weight gain after 9 weeks (range: 21.75.6 to 34.09.3g) did not differ significantly among
treatments (P>0.05). Survival rates were highly variable (40-75%) and not significantly different. Mortality
was not related to treatment. FCR (3.2 to 4.2) and PER (0.8 to 1.0) were not significantly different among
treatments.
Experiment 2. Feed peas in the diets ranged from 5.9-41%. Feed peas substituted up to 35% of the plant
protein (or up to about 30% of total dietary protein). The control diet contained fish meal, soybean meal,
and copra meal as principal protein sources. Separate feeding trials were conducted on two strains of all-
male tilapia (CLSU and BFAR strains) under controlled conditions using the same test diets and bigger
tanks (90x78x43 cm).
There were no significant differences in weight gain among treatments after 12 weeks of feeding (45.5-7.8
to 57.412.4g for the CLSU strain; 52.54.4 to 74.19.2g for the BFAR strain). Moreover, FCR (3.2 to 4.0)
or PER (0.9 to 1.0) were not significantly different among treatments. Survival was 100% in all tanks in
both trials.
2
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 of
the diets used in experiments 1 and 2 was also determined in separate feeding trials. The ADMD of the
diets 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) and
35% 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 feeding
trial. 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. Feed
pea meal was substituted at 0, 5, 10, 15, 20, 25, and 30% of total protein. A leading commercial milkfish
feed was also tested as an additional control. The experimental diets were fed to triplicate groups of
milkfish 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 the
commercial 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 feed
pea protein is well digested by milkfish.
Over-all results indicate that feed peas could be used as an alternate dietary protein source for milkfish
juveniles. 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 still
acceptable.
General Conclusions
These feeding trials clearly demonstrate that whole feed pea can be incorporated as an alternative protein
source 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 protein
ingredients, especially for fish that require high protein levels in their diet. In this case, feed pea can
therefore 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 studies
conducted. Feed peas contain high levels of lysine, which could complement the amino acids of other
ingredient 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 depending
upon 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 most
feed pea varieties now supplied in global markets. Thus, positive results were obtained for shrimp and
tilapia, even at high levels of incorporation in the diets. The relatively lower performance of milkfish
compared to shrimp and tilapia, even with feed pea comprising 15% of the diet, may be attributed to amino
acid deficiencies or possibly to effects from certain anti-nutritional factors. However, it is notable that the
growth response of milkfish fed the 15% feed pea diet was still better than from the commercial milkfish
diet, 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 open
avenues 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 energy
source for the animals.
3
Utilization Of Feed Pea, Pisum Sativum Meal, As An Alternative Protein Source
In Practical Diets For Juvenile Tiger Shrimp, Penaeus Monodon Fabricius
Myrna N. Bautista-Teruel, Perla S. Eusebio, and Timothy P. Welsh
Abstract
The 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 added
to the treatments. Twelve shrimp post-larvae with an average weight of 0.020.01g were randomly
assigned in 35, 60-l oval tanks equipped with a flow-through seawater system. The shrimps were fed the
formulated diets at a daily feeding rate of 20-25% body weight for 90 days in 5 replicate samples. No
significant differences (P<0.05) were observed in weight gain, feed intake, feed conversion ratio (FCR) and
protein efficiency ratio (PER) of shrimp fed diets with 0 up to the highest level of replacement. Weight gain
of shrimp fed the negative control was, however, significantly lower (P>0.05) compared to the rest of the
treatments. Specific growth rates (SGR) of shrimps for the various treatments showed the same trend as
the percent weight gain. Survival of shrimp for all treatments, including the negative control, was generally
high at 75-100%. The apparent dry matter (ADMD) and protein (APD) digestibilities of the dry feed pea in P.
monodon were 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. There
were no significant differences (P<0.05) in whole body composition (dry matter, protein, lipid, ash, fiber) of
shrimps fed the various diets with feed pea replacement. Pellet water stability was similar for all diets even
up to the highest level of replacement. The results have demonstrated that feed pea meal has very good
potential as a substitute protein source up to 100% of the protein from defatted soybean meal, which is
equivalent to 25% of total protein in the diet. An inclusion level of up to 42% in the juvenile shrimp P.
monodon practical diet did not manifest any adverse effects on growth, feed intake, FCR, survival, body
composition, and digestibility coefficients for dry matter and protein of the shrimp.
Keywords: Pisum sativum, Penaeus monodon, juvenile, nutrition, protein,
Introduction
The current trend in shrimp farming is toward increased intensification of culture systems. Success in this
type of culture is highly dependent on the availability of well-balanced, nutritionally complete and cost-
effective formulated feed. In shrimp feed formulation, soybean meal is considered the most widely used
plant protein source among the many available plant protein feedstuffs, owing to its nutritional quality,
favorable cost, and consistent availability (Akiyama, 1991). The utilization of soybean protein, however, in
human food and terrestrial animal feeds may bring about future problems concerning availability and
production cost. Thus, research should be conducted to assess the suitability of alternative protein sources
to soybean meal in formulated shrimp diets.
Feed pea (Pisum sativum) is a high energy, medium protein ingredient that has been widely and
successfully used for agricultural feeding in Europe, Canada, Australia, and the USA. These round-shaped,
low tannin varieties of Pisum sativum, are a major pulse crop marketed in these countries and globally as a
source of carbohydrates, mainly starch, and protein for livestock feeds (UNIP-ITCF, 1995). As a result of
plant breeding, several anti-nutritional factors in feed peas, including tannins and anti-trypsin, have been
eliminated or substantially reduced (Castell et al., 1996). The average protein content of feed pea is 22-
24% with a digestible energy (DE) of 3420 kcal/kg. Lysine is particularly high at 1.6%, or over 7% of
protein. Feed peas are easily handled, stored, and are processed well in compounded feeds.
Several reports have described feed peas as potential ingredients for aquaculture feeds (Allan, 1998;
Novoa and Castillo, 1998; Allan et al., 1999; Booth et al., 1999). Some reports have indicated that they are
acceptable ingredients in diets for rainbow trout (Kaushik et al., 1993; Gomes et al., 1993, 1995; Burel et al,
1996), silver perch (Allan, 1997; Allan et al., 1999; Booth et al, 1999), European seabass (Santos and
Gomes, 1997; Gouveia and Davies, 1998, 2000), and blue shrimp (Cruz-Suarez et al., 2001
This study evaluated the nutritive value of feed peas, Pisum sativum, as an ingredient for shrimp, Penaeus
monodon, practical diets. Effects of the dietary treatments on growth, survival, feed intake, apparent dry
matter digestibility, protein digestibility, feed conversion efficiency, and protein efficiency were investigated.
Water stability of the pellets prepared for the various treatments was also studied.
4
Materials and Methods
Experimental diets
The feed ingredients used in this study were obtained from commercial suppliers through the SEAFDEC
pilot 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 diet
formulation (Table 1). Six isonitrogenous diets were formulated to contain 40% protein (Table 2). Protein
from 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 (Kanazawa
and Teshima, 1981; NRC, 1993; FDS Manual, 1994; Millamena et al., 1996-1999). A negative control with
no protein sources was added to the treatments. Protein content of 40% in the shrimp diet has been
reported to be optimal for juvenile Penaeus 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, such
as fish meal, shrimp meal, shrimp head meal, and squid meal, were included in the dietary formulations at
the same levels. The levels of wheat flour and rice bran were adjusted to maintain similar dietary protein
contents. Diets were prepared according to standard diet preparation in the laboratory as described in FDS
Manual, 1994. To prevent lipid oxidation during storage, all diets were packed in sealed plastic containers
and 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 of
Official Analytical Chemists (AOAC 1990). The amino acid composition of both soybean meal and feed pea
meal and experimental diets was determined using the method of Simpson et al., 1976.
Feeding trials
Good quality and disease-free post-larval Penaeus monodon were sourced from a private hatchery in
Tigbauan, Iloilo, Philippines. They were placed in 2-tonne fiberglass tanks and fed with Artemia sp and
control diet for a week before they were transferred to experimental tanks. Twelve shrimp post-larvae with
an 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 days
in 5 replicate samples. Shrimp were fed three times daily at 0800, 1300, and 1700h. The wet weights of
the 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 for
proximate 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 methods
of AOAC (1990).
Biological analysis
Diet performance was evaluated by calculation of percent weight gain=final weight-initial weight/initial
weight x 100; specific growth rate=100 (ln ave. final wt-ln ave. initial wt)/no. of culture days; feed conversion
ratio (FCR)= total dry feed intake (g)/wet weight gain (g); percent survival =final number of shrimp/initial
number of shrimp x 100.
In vivo digestibility experiment
Apparent digestibility coefficients for dry matter (ADMD) and crude protein (APD) of feed peas as ingredient
and feed-peas based diets were measured using 1% Cr
2
O
3
as external indicator. The method by Cho. et
al. (1982) was adapted in a ratio of 70:30 (reference diet to test ingredient) in each test diet. The control
diet 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 body
weight of 14.53 0.57g and 8 shrimp with mean body weight of 13.560.56g were stocked in 250 L conical
fiberglass tanks for the test ingredient and feed pea-based diets, respectively. The shrimp were acclimated
to 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 and
flow rate of 600-700 ml/min was provided for each tank. Water temperature and salinity were maintained at
24-48C 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 a
collecting vial. Care was taken to prevent the breaking-up of fecal strands to facilitate collection and to
avoid leaching of nutrients. Thereafter, fecal samples were carefully rinsed with distilled water and
immediately stored in a freezer to retard bacterial decomposition. At the end of fecal collection, samples
5
were thawed and pooled prior to freeze-drying. Dried fecal samples were thoroughly mixed and prepared
for chromic (Cr
2
O
3
) (Carter et al., 1960) and crude protein analysis (AOAC, 1990). Apparent dry matter
digestibility (ADMD) and apparent protein digestibility (APD) were determined using the equation:
% Digestibility = 100-100 X [{C
d
/C
f
} X {N
f
/N
d
}] where: C
d
= % chromic oxide in diet; C
f
= % chromic oxide in
feces; N
f
= % nutrient in feces; N
d
= % nutrient in diet
Water stability test for diets
Water stability of the diets was determined at 4, 8, 12, 16, and 24 h following the method described in FDS
Manual, 1994. Percent water stability was computed as final dry weight of feed / initial dry weight of feed X
100.
Statistical analysis
All data were analyzed by a one-way ANOVA using a Statistical Analysis Software Program of SPSS. The
Duncans 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<0.05.
Results
Proximate composition of the experimental diets is shown in Table 4. All diets were isonitogenous, with
protein content of 40% and isolipidic containing 7-8% lipid. Amino acid analysis of the experimental diets
(Table 5) showed conformity with the amino acid requirements of shrimp (Kanazawa and Teshima, 1981;
NRC, 1993; FDS Manual 1994; Millamena et al., 1996-1999). Results of the feeding trial are summarized
in Table 6. No significant differences (P<0.05) were observed in weight gain, feed intake, feed conversion
ratio (FCR) and protein efficiency ratio (PER) of shrimp fed diets with 0 up to the highest level of
replacement (100%). Weight gain of shrimp fed the negative control was however, significantly lower
(P>0.05) compared to the rest of the treatments. Specific growth rates (SGR) of shrimp fed the various
treatments showed the same trend as the percent weight gain. Survival of shrimp for all treatments,
including the negative control was generally high at 75-100%. The apparent dry matter (ADMD) and protein
(APD) digestibilities of the dry feed pea in P. monodon were high at 73.384.98 and 92.742.62,
respectively (Table 7). In the case of the feed pea meal-based diets, Diet 4 with 80% level of replacement
had the highest apparent digestibility coefficients (ADC) for dry matter (ADMD) at 77.33, which was
significantly higher (P<0.05) than diets with 0, 20, and 60% replacement (Table 8). The apparent protein
digestibility for protein (APD) in diet 4 with 80% replacement was significantly high at 87.52. This value was
however, not significantly different for the APD of diet 3 with 60% replacement (85.69). The feed pea
replacement level did not affect body composition of the shrimp, (protein at 62-72%; fat at 3.0-3.5%; ash at
19-24%; fiber at 2-3%) even after the termination of the feeding trial (Table 9).
The data on pellet water stability of various diets measured at 4, 8, 12, 16, 24 h are presented in Table 10.
Percent recovery of experimental diets range between 80 to 91% after 24 h. Values were generally high for
all treatments even after the longest hour of submersion into seawater.
Discussion
The non-significant differences in growth rate of shrimp from 0 to highest level of feed pea replacement in
the diet may indicate that whole raw feed pea can be utilized even at the highest level of total protein
replacement at 25%. All the prepared diets, even up to the highest level of feed pea replacement, were
highly acceptable and were noted to be consumed readily by the shrimp. Aside from the replaced
ingredient, all the formulated diets must have contained the necessary attractants that the shrimp require,
thus high acceptability of all the diets was noted.
Results of this study have shown that total substitution of soybean meal with raw feed peas did not affect
shrimp performance. These results conform to the reports of others, and as cited in the introduction, feed
pea meal can be successfully included in practical aquaculture diets. When alternate plant protein sources
are used in diets containing the same concentration of digestible energy and protein and are able to meet
the nutritional requirements of the animal being fed, similar performance may be expected (Cruz-Suarez et
al., 2001). Since all the diets were isonitrogenous, at 40%, the quality of protein may make the difference.
Amino acid composition of the diets tested have shown that the requirements of the shrimp for these
nutrients have been met in all the dietary treatments, thus growth of the shrimp was similar. High
digestibility of the feed pea in shrimp, as shown in the digestibility coefficients for dry matter and protein
obtained in this study, may be another contributing factor in the good growth of shrimp fed diets with various
levels of feed pea replacement. The increasing level of digestibility of the diets in the shrimp with an
increase in the level of feed pea replacement may suggest that, even at the highest level of soybean
replacement, feed pea is still highly digestible.
6
Smith et al. (1999) reported that digestibility coefficients of feed peas in tiger shrimp were 80% for ADMD
and 91% for APD. The ADMD value and APD value is almost the same as those obtained in this present
study (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 feed
peas 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 et
al., 1993) and European sea bass (Giuveia and Davies, 2000) have emphasized the requirement for
dehulling and extrusion cooking. Eusebio (1991) also found that processing of cowpea and rice bean had
no significant effect on growth and survival of P. monodon, although apparent protein digestibility of rice
bean was improved. Shrimp are more tolerant than other species of certain anti-nutritional factors in the
seed coat components (Cruz-Suarez et al., 2001).
This study has demonstrated the acceptable nutritional value of whole feed pea as an ingredient in shrimp
diets, 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 up
to 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 Moisture
Crude
Protein
(N x 6.25)
Crude fat
Crude
fiber
NFE
c
Ash
Feed pea
a
11.58 25.24 1.32 3.68 65.96 3.80
Soybean meal
b
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 27.78
Rice bran 10.12 14.22 18.40 7.20 50.17 10.01
c
NFE-Nitrogen Free Extract
*Means of 2 replicate samples
a
Whole feed pea
b
Defatted soybean meal
Ingredients 1 2 3 4 5 6 7
% Soybean replacement 0 20 40 60 80 100
Neg.
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; dicalcium
phosphate,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, 7.20g
kg
-1
; pyridoxine, 6.60g kg
-1
; cyanocobalamine, 0.02gm kg
-1
; _-tocopherol, 16.50 gm kg
-1
; menadione, 2.40
gm 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%; Zn
0.42%; Cu, 0.21%; K, 7.50%; Co, 0.011%; Mn, 0.160%; Se, 0.001%; Mo, 0.0005%; Al ,0.0025%; I, 0.04%.
7
Table 3 Composition of reference and test diets for in vivo digestibility experiment in shrimp,
P. monodon juveniles (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
diets
a
Feed Ingredient
Reference
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
Cr
2
O
3
1.00 1.00
Feed peas - 30.00
* Phosphitan C, feed grade, Showa Denko K.K. Japan
Diet No. Moisture
Crude
Protein
Crude Fat
Crude
Fiber
Nitrogen
Free
Extract
Ash
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
Methionine
b
0.63 0.19 0.27 0.29 0.31 0.33
Threonine
b
1.72 1.06 0.84 0.86 0.83 0.86
Serine 2.46 1.33 0.91 0.94 0.95 0.97
Glutamic 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
Valine
b
1.41 1.26 1.02 1.04 1.05 1.08
Isoleucine
b
1.42 1.22 1.22 1.23 1.23 1.71
Leucine
b
4.07 2.2 1.93 1.95 1.96 1.97
Tyrosine
b
1.84 0.9 0.87 0.88 0.89 0.89
Phenylalanine
b
1.93 1.32 0.96 0.98 1.09 1.05
Lysine
b
2.73 2 1.84 1.86 1.84 1.89
Histidine
b
1.05 0.57 0.54 0.54 0.53 0.55
Arginine
b
2.56 1.9 1.45 1.57 1.69 1.68
a
Values are mean of two replicate samples
b
Essential amino acids for shrimp, Penaeus monodon (Kanazawa and
Teshima, 1981; NRC, 1993; FDS Manual 1994; Millamena et al., 1996-1999)
Amino acid
Soybean
meal
Feed pea
meal
% Replacement in the diet
8
Table 6 Response of juvenile P. monodon over a 90-d feeding trial to experimental diets containing
replacement 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 Penaeus
monodon (%).
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.
Moisture
Crude
Protein
(N x 6.25)
Crude Fat
Crude
Fiber
Nitrogen
Free
Extract
a
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 samples
a
NFE-Nitrogen Free Extract
Dietary
treatments
% ADMD %APD
Diet 1 (0)
73.40
b
82.76
c
Diet 2 (20)
70.58
b
84.40
bc
Diet 3 (60)
71.82
b
85.69
ab
Diet 4 (80)
77.33
a
87.52
a
Nutrient digestibility (%)=100-
(100x%Cr
2
O
3
diet/%Cr
2
O
3
feces x
%nutrient feces/%nutrient diet)
Diet No.
Initial mean
wt (g)
Final mean
wt (g)
Weight
gain
a
(%)
SGR
b
FCR
c
PER
d Survival
e
(%)
1 (0) 0.020.01 1.300.08 5598309
a
4.490.06
a
1.230.14
a
1.230.11
a
83
2 (20) 0.020.01 1.130.06 516292
a
4.400.01
a
1.770.04
a
1.440.03
a
100
3 (40) 0.020.01 1.450.04 5980165
a
4.610.01
a
1.360.12
a
1.430.01
a
75
4 (60) 0.020.01 1.370.01 583965
a
4.540.12
a
1.890.03
a
1.350.02
a
92
5 (80) 0.020.01 1.210.18 5187395
a
4.380.15
a
1.390.32
a
1.340.08
a
100
6 (100) 0.020.01 1.190.07 5223205
a
4.410.04
a
1.390.33
a
1.660.08
a
100
7 (neg) 0.020.01 0.100.02 36482
b
1.670.20
b
7.021.86
b
0.110.01
b
75
d
PER = weight gain (g) protein intake
e
Survival (%) = 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 significantly
different (P<0.05)
a
Weight gain (%) = (final wt-initial wt)/initial wt x 100
b
SGR = 100 (ln ave. final wt-ln ave. initial wt)/number of days
c
FCR = dry feed intake/wet weight gain
Ingredient ADMD APD
Feed Pea 73.384.98* 89.09 1.96*
92.742.62**
Nutrient digestibility (%) = 100-(100 x%Cr
2
O
3
diet/%Cr
2
O
3
feces x %nutrient feces/%nutrient
*Cho et al ., 1982
**Foster, 1999
9
Table 10 Percent recovery of experimental diets in seawater*
Acknowledgement
The authors acknowledge the United States Department of Agriculture and the USA Dry Pea and Lentil
Council for the research and travel funding support; M. Mallare and J. Vera Cruz for the technical
assistance; F. Jarder for the proximate analysis; J. Bangcaya and M. Arnaiz for the amino acid analysis,
and M.J. Bernas for the Cr
2
O
3
analysis.
References:
Akiyama, D.M. 1991. Soybean meal utilization by marine shrimp. In: D.M. Akiyama and R.K.H. Tan (eds). Proceedings of the
Aquaculture Feed Processing and Nutrition Workshop. Thailand and Indonesia. September 19-25, 1991. American Soybean
Association. Singapore. pp 207-225.
Alava, V. R. and Lim, C. 1983. The quantitative dietary protein requirement of Penaeus monodon juveniles in a controlled
environment. Aquaculture. 53:229-242
Allan, G., Stone, D.A.J., Booth, M.A., 1999. Alternative protein sources: plant proteins. Book of abstracts of the World Aquaculture
Society Annual Meeting 99. April 26-May 2, 1999. Sydney, Australia. WAS, Baton Rouge. I.A. USA, 18.
Allan, G. 1998. Protein sources.potential for pulses. .International Aquafeed 2:.17-20
Allan, G.L. ,Smith, D.M. 1998. Recent Nutrition Research with Australian Penaeid. Reviews in Fisheries Science. pp. 113-127.
Allan, G., 1997. Potential for pulses in aquaculture systems. Proceedings of International Food Legume Research Conference III,
Sept. 22-26, 1997, Adelaide, Australia. 13 pp.
AOAC (Association of Official Analytical Chemists), 1990. Official methods of analysis, 15
th
edition. Association of Official Analytical
Chemists. Arlington, Virginia, USA.
Bautista, M.N. 1986. The response of Penaeus monodon juverniles to varying protein/energy ratio in test diets.
.Aquaculture.,53:229-242
Booth, M.A., Allan, G.L., Stone, D.A.J., 1999. Utilization of four agricultural aingredients by silver perch. Book of abstracts of the World
Aquacultue Society Annual Meeting 99, April 26-May 2, 1999. Sydney, Australia. WAS, Baton Rouge. I.A. USA, 20.
Burel, C., Boujard, T., Boeuf. G., Evrard, J., Peyronnet, C., Kaushik, S.J., 1996. Utilisation de proteins dorigine vegetale (pois, lupin,
colza) dans lalimentation de la truite arc-en-ciel: valeur nutritionelle et effets sur laxe thyreotrope. In: CRITT Valicentre (Eds),
Proceedings of Colloque Annuel Valicentre, Ardon, France, 28 Nov. 1996, pp 47-58.
Carter, J.F., Bolin, D.W., Erikson, P., 1960. The evaluation of forages by the Agronomic difference method and chromogen chromic
oxide indicator technique. N.D. Agric.
Castell, A.G., Guenter, W., Igbasan, F.A. 1996. Nutritive values of peas for non-ruminant diets Animal Feed ScienceTechnology.
60:209-227
Cho, C.Y., Slinger, S.J., Bayly, M.S., 1982. Bioenrgetics of salmonid fishes; enrgy intake, expenditure and productivity. Comp.
Biochem. Physiol. 73B, 25-41.
Cruz-Ricque, L.E. Guilllaume, J. Cuzon, G. AQUACOP, Taravao ( French Polynesia) 1987. Squid protein prfotien effect on growth
of four penaeid shrimp. 16 ref. Journal of the world Aquaculture Society. Baton Rouge L.A. Vol 18,No.4. pp209-217
Cruz-Suarez, L.E., Ricque-Marie, D., Tapia-Salazar, M., McCallum, I.M., Hickling, D., 2001. assessment of differently processed feed
pea (Pisum sativum) meals and canola meal (Brassica sp.) in diets for blue shrimp ( Litopenaues stylirostris). Aquaculture 196,
87-101.
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, Penaeus
monodon juveniles. Aquaculture 99, 297-308.
Feed Development Section, 1994. Feeds and feeding of Milkfish, Nile Tilapia, Asian sea bass, and Tiger Shrimp. SEAFDEC
Aquaculture 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) on
growth, 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 (Oncorhynchis
mykiss). 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 (Dicentrarchus
labrax). 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 85
Diet 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
10
Kaushik, S.J., Vachot, C., Aguirre, P., 1993. Potential utilization of extruded peas. 6
th
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, Penaues
monodon (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 Penaeus
monodon. 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 by
different 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 the
World 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 (Dicentrarchus
labrax). 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 des
Plantes Riches en Proteines, Paris, France, 99 pp.
11
Feed Pea (Pisum Sativum) As An Alternative Protein Source
In The Diets Of Nile Tilapia, Oreochromis Niloticus (L.)
Corazon B. Santiago, Perla S. Eusebio, and Timothy P. Welsh
Abstract
A study was conducted to test feed pea (Pisum sativum) as a protein source in Nile tilapia diets and to
determine 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 not
contain peas. Manually sexed male Nile tilapia, weighing 32-39 g per fish at stocking, were used. The fish
were reared in polyethylene tanks (58x37x27 cm) with aeration. Water was static but was partially changed
daily. Water temperature for the whole duration of the experiment ranged from 22-27C. Fish were allowed
to 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) did
not differ significantly among treatments (P>0.05). Survival rates were highly variable (40-75%) and
mortality was not related to treatment. For experiment 2, plant-based tilapia diets were also designed to be
isonitrogenous and isocaloric. Feed peas in the diets ranged from 5.9-41.0%. Feed peas substituted up to
35% of the plant protein (or up to 29% of total dietary protein). Separate feeding trials were conducted on
two 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 the
CLSU strain; 52.5+4.4-74.1+9.2 g for the BFAR strain). FCR and PER were not significantly different
among treatments. Survival was 100% in all tanks in both trials. Determined with the use of chromic oxide
as a marker, the apparent protein digestibility of feed pea as ingredient was 87.2+2.0%. Diets in
experiments 1 and 2 that contained peas had more than 90% protein digestibility. Overall, feed pea is an
alternative protein source that can be used routinely in the diets of Nile tilapia.
Key words: Tilapia nutrition; Peas; Alternative protein source; Fishmeal substitute; Digestibility
Introduction
Fish meal is a major source of highly digestible nutrients and is a palatable ingredient in fish diets. It can be
the 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) is
increasing while the supply is stagnating or even decreasing (Starkey, 1994). This causes the rising cost of
fish meal. In view of the need to minimize dependence of aquaculture feeds on fish meal, the search for
alternative protein sources has been an international concern.
The most commonly used plant protein sources that could partially substitute fish meal in fish diets are oil
seed meals, some leaf meals, and cereals and their by-products (Gerpacio and Castillo, 1979; Zamora and
Baguio, 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). The
grain legumes are also called pulses, which refer to edible seeds of plants with pods belonging to the family
Leguminocae (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 (Bidyanus
bidyanus) (Allan et al., 2000). Feed pea (Pisum sativum), in particular, has been tested to replace fish meal
in the diets of rainbow trout (Gouveia et al., 1993), European sea bass (Dicentrarchus labrax) (Gouveia and
Davies, 1998), and Atlantic salmon (Salmo salar) (Carter and Hauler, 2000). Because of its high potential
as a fish feed ingredient, feed pea has to be tested in other important food fish, including tilapia. Thus, the
present study was conducted to determine the effect of levels of dietary feed peas on the growth and
survival of juvenile Nile tilapia and on feed efficiency. The digestibility of feed peas as an ingredient and of
diets containing feed peas was also determined.
Materials and Methods
1. 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
12
exceeded amino acid requirements of young Nile tilapia (Santiago and Lovell, 1988). A fishmeal-based diet
allowed the incorporation of high levels of feed peas in the test diets. The feed peas (12.7-63.3% of the
diet) substituted 10-50% of fishmeal protein, or about 9-48% of total dietary protein. The diet without feed
peas served as a control. Because of the high nitrogen-free extract (NFE) in feed peas, the diets had about
the same energy content, but the supplemental starch and oil decreased as the feed pea in the diet
increased. Based on the actual analysis of protein, lipid, and NFE, however, the digestible energy of the
diet 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 were
also 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 from
soybean 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 peas
contained 22.32% crude protein, 1.17% crude fat, 3.36% ash, 3.25% crude fiber, and 58.32% nitrogen-free
extract. These values are close to those reported for feed peas (Muehlbauer and Tullu, 1997; Pulse
Canada, 1999a; Racz, 1999).
2. Experimental fish and tanks
Three strains of tilapia juveniles were obtained for the study. The young fish were reared in the laboratory
for 6-8 weeks before use. During this time, the fish were fed dry pellets without feed peas. Each batch of
fish 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 the
Station. Each fish weighed 32-39 g at stocking. Twenty-four polyethylene tanks measuring 58x37x27 cm
were 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 strain
individually 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 feeding
trial, 16 tanks were randomly stocked with six juveniles (BFAR strain) per tank.
3. Feeding treatments, sampling and water management
Each of the feeding experiments was conducted in a completely randomized design. In experiment 1, there
were six dietary treatments with four replicates each. The treatments represented levels of fishmeal protein
substitution by the feed peas. In each of the two feeding trials in experiment 2, there were eight treatments
or levels of plant protein substitution by feed peas. Each of the dietary treatments had three replicates for
the 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 in
experiment 2. The amount of feed given to the fish at each feeding exceeded satiation level to ensure that
the 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 was
back 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 the
succeeding weeks except in experiment 1 when the final measurements were done after 9 weeks. Total
length of fish was measured at stocking and during harvest. Feed conversion ratio (FCR, the amount of
feed consumed divided by weight gain of fish), protein efficiency ratio (PER, weight gain divided by weight
of 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 static
but 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 every
morning before water was partially changed with the use of YSI DO meter (Model 55/25 FT). The pH was
measured with a pH meter (Beckman, Model PHI 50). Total ammonia nitrogen (NH
3
-N) was determined
weekly before changing of water in the morning by the Nessler method with the use of a Hach kit
(DREL/2010).
13
Water temperature for the whole duration of experiment 1 ranged from 22-27C, being at the lower range
for most of the days. Dissolved oxygen in the morning before partial water change ranged from 3.5-6.2 mg l
-
1
. The total NH
3
-N ranged from 0.63-2.39 mg l
-1
and 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 NH
3
-N, 0.21-2.42 mg l
-1
. For the
feeding trial using the BFAR strain, water temperature also ranged from 23-27C; pH, 7.6-8.7; and total
NH
3
-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 were
obtained for proximate analysis by standard methods (AOAC, 1990). At the end of the each feeding trial
and after a 24-hour fasting, fish samples were obtained and dissected. The appearance of the fish and the
internal organs were examined visually for any abnormality. Other samples of fish that received the diets
whose digestibility was determined were subjected to cold shock, weighed and kept frozen until they were
processed further. The fish samples were then autoclaved, homogenized, dried in a vacuum oven at 60C,
and ground for analysis.
5. Digestibility measurements
5.1. Digestibility of feed peas as ingredient
The apparent dry matter digestibility (ADMD) and crude protein digestibility (APD) of feed peas as
ingredient were measured indirectly using chromic oxide (Cr
2
O
3
) as external indicator (Cho et al., 1982;
Spyridakis et al., 1989). The reference diet for the measurement of digestibility of feed peas was the same
as the control diet used in experiment 1 (Table 1), except that 1% Cr
2
O
3
and 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% Cr
2
O
3
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 the
water 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 25
consecutive days. Fecal material were carefully rinsed with distilled water and stored in a freezer to retard
bacterial decomposition. Samples were pooled and freeze-dried. Dry fecal samples and test diets were
analyzed for Cr
2
O
3
(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 diets
with 0, 15, 25 and 35% replacement of plant protein by feed peas (Table 2), was determined in tilapia in
separate trials. Celufil (filler), starch, and rice bran were adjusted to accommodate 1% Cr
2
O
3
and the binder
in 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 the
same as in the previous digestibility trial for peas as an ingredient. The apparent digestibility for dry matter
and 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 protein
and ash, and digestibility coefficients were analyzed by ANOVA using the General Linear Models
Procedure by SAS for PC (SAS Institute Inc., 1991). When significant difference among treatments was
detected at P<0.05, Duncan's multiple range test was used to determine which means were different from
each other.
Results
The weekly or biweekly body weight of the Nile tilapia juveniles over a 9-week period in experiment 1 did
not show significant differences among treatments (P>0.05). Similarly, weight gain and increase in total
length of the Nile tilapia after 9 weeks showed no significant differences among treatments (Table 3).
14
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 not
differ significantly among treatments (P>0.05). Mortality was caused by the aggressive behavior of the male
tilapia 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 differ
significantly 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 BFAR
strain of tilapia in experiment 2 did not show significant differences in body weight during each sampling, or
weight gain and total length increment over a 12-week period (Table 5). Moreover, the FCR and PER did
not 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% feed
peas (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. In
experiment 2, carcass protein as well as body ash differed significantly among treatments (Table 6), but no
trend could be established in relation to fish growth and digestibility of the diets. However, as the dietary
feed peas in the plant-based diets increased and the dietary lipid decreased (Table 2), carcass lipid
decreased (Table 6).
There was no visible adverse effect of dietary peas on feeding behavior and the appearance of the fish and
the internal organs. The three tilapia strains used in the feeding trials readily accepted the diets containing
feed peas and consumed almost the same amount of feed in relation to their body weight (about 5% of fish
biomass). Visual examination at the end of each trial revealed that the tilapia were generally lean. Fish liver
did 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 protein
digestibility (APD) was 87.2+2.0%. The ADMD coefficients for some fishmeal-based diets used in
experiment 1 significantly increased with increasing level of feed peas (Table 7). The APD for the diets
tested 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 diets
in which digestibility coefficients were determined also showed a significant increase in ADMD when feed
pea 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.
Discussion
The three feeding trials consistently demonstrated that feed peas could be incorporated in tilapia diets at a
wide range of levels to replace fishmeal protein or other plant proteins without affecting fish growth, feed
efficiency, and survival. The FCR values obtained for all diets in the feeding trials were higher than the
expected values (about 2) and, consequently, the PER values were fairly low. These could be attributed to
the 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 system
had 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 feeding
trials had 100% survival. The growth response of tilapia to diets with peas in the different feeding trials was
not 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 was
observed in fish fed plant-based diets, but not in fish fed the fishmeal-based diets. Moreover, carcass lipid
of the tilapia (CLSU strain) fed plant-based diets decreased with the increase of feed peas in the diet. In a
companion study, feeding of milkfish (Chanos chanos) with varying dietary feed peas caused significant
differences in growth but it did not influence carcass composition (I. Borlongan, personal communication
2002). Similarly, two levels of peas in the diets (20 and 40%) did not affect carcass protein and ash in
European 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 be
incorporated in practical applications will be influenced largely by the cost of the peas in relation to the cost
15
of competing protein sources. The digestibility of the peas is also considered in evaluating its potential as a
protein source. In the present study, the digestibility coefficients for feed peas in Nile tilapia are comparable
to those reported for other freshwater fishes (Table 8). The highest digestibility coefficients for P. sativum
were obtained in turbot, a marine species. In a companion study, the crude protein digestibility of feed peas
in 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 ingredients
in silver perch as well as that of amino acids in the ingredients. The protein digestibility of the ingredients
was 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 amounts
of carbohydrate than lipid (Muehlbauer and Tullu, 1997; Pulse Canada, 1999a; Racz, 1999) and the protein
digestibility coefficients for peas in different fish species (Table 8) are close. Moreover, the apparent energy
digestibility coefficients reported for peas as ingredient are near the values for the carbohydrate digestibility
of 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. sativum in fish diets is limited. Gomes et al. (1995) used feed pea at very a low
level (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 a
study by Gouveia et al. (1993), pea seed meal was used at 38.2% of the diet (20% of the dietary protein) of
rainbow trout with or without cooking/expansion (145

C, 25 kg per cm
2
). The thermal treatment slightly
improved the nutritional value of the diets containing the peas. In another study, there was a significant
decrease in the protein digestibility of the diet when the pea (P. sativum) in the diet of rainbow trout
increased from 2550% (Pfeffer et al., 1995). Furthermore, the digestibility of the diets containing either 25
or 50% peas increased significantly when the peas were autoclaved. APD coefficients of rainbow trout diets
containing 25% feed peas increased from 86-91% when the peas had pre-treatment. For diets containing
50% peas, APD increased from 83-86% when the peas were autoclaved (Pfeffer et al., 1995). In a
subsequent study by Gouveia et al. (1998) on European sea bass (D. labrax), fishmeal-based diets
containing 20 and 40% pea had APD coefficients of 88-89%. In the present study, the APD coefficients of
the tilapia diets containing feed pea were high (90.5-92%) and were practically not affected by the dietary
level 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 most
likely be higher in tilapia since dehulling removes most of the fiber of ingredients (Eusebio, 1991). When
compared to other feedstuffs tested in tilapia, feed pea has APD coefficient that is comparable to that of fish
meal (Hanley, 1987; NRC, 1993; Jauncey, 1998). However, it is slightly lower than the protein digestibility
values 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 fish
meal 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 a
practical 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 in
lysine (e.g. canola meal and gluten meals). However, as in soybean meal and other legume seeds, the
sulfur amino acids (methionine and cystine) are low and could be the first limiting amino acids when high
amounts 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. Cow
pea (Vigna unguiculata) protein concentrate was also tested in Nile tilapia fry and up to 40% inclusion level
in 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-nutritional
factors (Tacon, 1987; Hertrampf and Piedad-Pascual, 2000). However, through genetic selection, present
pea 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), unlike
soybean 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.
16
The consistent response of the tilapia juveniles to diets containing feed peas in three feeding trials clearly
showed that feed pea as an alternative ingredient can be routinely included in tilapia diets. Further studies
on feed peas as component of tilapia diets (e.g., those related to processing and energy utilization) have to
be 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 Nile
tilapia fed diets containing feed peas at varying fishmeal protein substitution for 9 weeks
(experiment 1)
a
Ingredient 0
a
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 13.9
a
Diet 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)
a
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)
a
Diet 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+0.1
40 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)
a
Column means are not significantly different from each other (P>0.05). Mean+SEM
Fishmeal protein
substitution (%)
Dietary feed
peas (%)
FCR
Survival
(%)
PER
Body weight (g)
17
Table 4 Mean initial weight and total length, gain in weight and length, FCR and PER of tilapia (CLSU
strain) 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 2
a
Initial Gain Initial Gain
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+0.1
15 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
a
Column means are not significantly different from each other (P>0.05). Mean+SEM
Total length (cm) Dietary feed
peas (%)
Plant protein
substitution (%)
FCR PER
Body weight (g)
Initial Gain Initial Gain
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
a
Column means are not significantly different from each other (P>0.05). Mean+SEM
Total length (cm)
Plant protein
substitution 0.80
Dietary feed
peas (%)
Body weight (g)
Crude
protein
Ash Lipid
Diets in Expt 1
0 - 56.5 0.2
a
18.9 0.1
b
18.80.3
b
10 12.7 56.4 0.6
a
19.5 0.3
b
19.40.0
a
30 37.5 57.2+0.4
a
20.8+0.4
a
18.70.0
b
50 63.3 57.4+0.2
a
20.8+0.1
a
18.90.1
b
Diets in Expt 2
b
0 - 55.7+0.4
c
16.3+0.6
b
23.20.4
a
15 17.6 59.1+0.4
a
14.7+0.2
c
21.20.3
b
25 29.3 56.5+0.2
c
16.3+0.3
b
20.90.3
b
35 41.0 58.2+0.2
b
18.3+0.0
a
20.30.1
b
a
For each experiment, column means with a common superscript are not
significantly different (P>0.05).
b
At 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
Dietary feed
peas (%)
% fishmeal protein substitution
% plant protein substitution
18
Table 7 Apparent dry matter digestibility (ADMD) and protein digestibility (APD) of diets with peas in
juvenile male tilapia (CLSU strain)
Table 8 Digestibility coefficients (%) for P. sativum in different fish species
Acknowledgements
This study was funded by the US Department of Agriculture and sponsored by the USA Dry Pea and Lentil
Council. Feeds and fish samples were analyzed by the staff of the Central Analytical Laboratory of
SEAFDEC AQD, Tigbauan, Iloilo or the Animal Nutrition Analytical Service Laboratory, University of the
Philippines Los Baos, Laguna.
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(Dicentrarchus labrax). Aquaculture 166, 311-320.
Digestibility
trials
Dietary feed
peas (%)
ADMD (%)
a
APD (%)
a
0 0 72.1 0.6
c
92.5 0.4
a
10 12.7 75.6 0.4
b
90.5 0.4
b
30 37.5 77.8+1.1
ab
91.5+0.7
ab
50 63.3 78.8+0.6
a
92.0+0.2
ab
0 0 74.1+0.2
b
90.6+0.3
a
15 17.6 73.4+0.59
b
91.5+0.3
a
25 29.3 76.2+0.67
a
91.2+0.3
a
35 41 76.9+0.42
a
91.6+0.3
a
Trial using diets in Expt 1
Trial using diets in Expt 2
a
For 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
19
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, The
Netherlands, 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, EAAP
Publication, 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 or
hydrothermically treated legume seeds in diets for rainbow trout, Oncorhynchus mykiss (Walbaum), on apparent digestibilities of
their 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.
20
Potential Of Feed Pea (Pisum Sativum) As An Alternative Protein Source
In Practical Diets For Milkfish (Chanos Chanos Forsskal)
Ilda G. Borlongan, Perla S. Eusebio, and Timothy P. Welsh
Abstract
A 12-week feeding trial was conducted to evaluate the use of feed pea as a potential dietary protein source
for juvenile milkfish, Chanos chanos Forsskal. Six isonitrogenous (30% crude protein) and isocaloric (3940
kcal/kg) practical diets were formulated. The control diet contained fish meal, soybean meal, meat and
bone 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 additional
control. The experimental diets were fed to triplicate groups of milkfish fingerlings (mean initial weight of
0.42 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 fish
fed the control diet. Replacement with feed pea at 15% and higher levels led to milkfish fed these diets
showing 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 with
feed pea showed better growth rates and feed conversion ratios than the commercial feed control. Whole
body composition (crude protein, crude fat, crude fiber, nitrogen free extracts, and ash content) of milkfish
fed 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 protein
source 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)
Introduction
The current trend in milkfish culture is toward increased intensification of culture systems whereby provision
of 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 done
considerable research on the nutrient requirement of milkfish, assessment of nutritive value of available
ingredients, and development of simple and appropriate feeding technology. These are all important
factors 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 feeds
to 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 the
search for alternative protein sources for fish feed to sustain fish production. The use of plant protein
sources to completely or partially replace fish meal in fish diets has been studied for many years by several
workers 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 et al., 1997;
Watanabe et al., 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 high
energy, 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 of
this ingredient as dietary protein source for aquaculture species.
The aim of the current study was to assess the potential of feed pea (Pisum sativum) as an ingredient in
practical feeds for milkfish (Chanos chanos Forsskal). This was compared with solvent-extracted soybean
meal because soybean meal is the most often used plant alternative to fish meal in milkfish feeds. The first
phase of the experiment was conducted to establish the nutritive value of feed pea through measurement of
fish growth, feed utilization and carcass composition. The second phase assessed the apparent
digestibility coefficients for dry matter and dietary protein.
Materials and Methods
Experiment 1: Growth performance and feed utilization
Experimental Diets
The test diets were formulated to be isonitrogenous and isocaloric. The control diets were the existing
SEAFDEC 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
21
experimental diets are presented in Table 1. All diets were formulated to contain 30% protein, 10% lipid
and 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. The
basal formulation contained fish meal, defatted soybean meal, meat and bone meal, and copra meal as
dietary protein sources. The levels of feed pea incorporation tested were 0, 5, 10, 15, 20, 25, and 30% of
the total crude protein. Dietary essential amino acid profiles (Table 2) were calculated based on analyses
of 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 and
soybean 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 pea
was oven-dried at 60
0
C 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 by
cooking in 600 ml water and added to the mixture. The semi-moist mixture was then passed through the
Hobart food grinder to form 2 mm diameter pellets. The pellets were dried in an air convection oven at
40
0
C. The dry pellets were then ground, sieved to uniform sizes, and stored at 4
0
C until used for feeding.
Experimental fish and feeding management
The experiment was conducted at the Feed Development Section of the Aquaculture Department of the
Southeast 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 (control
diet) prior to the experiment. The feeding trials were conducted in an indoor flow-through system. Sixty-liter
capacity, oval fiberglass tanks containing 50L seawater with aeration were used. Milkfish juveniles (mean
wt. = 0.42 g) were randomly distributed at a stocking rate of 10 fish per tank and in four replicate tanks per
treatment. Fish in each treatment were then fed three times daily at 0830, 1130, and 1400H at a feeding
rate of 10% body weight per day for 12 weeks. Feeding ration was adjusted at every 3 weeks sampling
interval. Water quality parameters (temperature, salinity, DO, pH, ammonia, nitrite) were monitored
following the methods of Strickland and Parsons (1972) to ensure water quality remained well within limits
recommended for milkfish culture (Bagarinao, 1991).
Statistical Analyses
Survival, % weight gain, specific growth rate (SGR), food conversion ratio (FCR), and protein efficiency
ratio (PER) were calculated and subjected to statistical analyses. Data were subjected to analysis of
variance (ANOVA) and Duncans Multiple Range test (DMRT). Differences between means were
considered significant at P<0.05. All statistical tests were performed using the SAS/STAT Software
Program for Windows.
Chemical Analyses
The proximate composition of the dietary ingredients were determined prior to diet formulation and that of
the experimental diets after preparation and prior to feeding experiments according to AOAC (1990)
methods. Fish carcass composition after feeding was also determined.
The amino acid composition of feed pea and experimental diets were analyzed using high performance
liquid chromatography (HPLC) in a Shimadzu Liquid Chromatograph Model LC-10AT. The samples were
hydrolyzed using methane sulfonic acid (MSA) hydrolysis method according to Simpson et al., 1976.
Experiment 2: Digestibility Determination
Apparent digestibility coefficients for dry matter (ADMD) and crude protein (APD) of feed pea and
experimental diets in milkfish were determined using 1% chromic oxide (Cr
2
O
3
) as external indicator and a
modified Guelph fecal collection system (Cho et al., 1982; Eusebio and Coloso, 2000) for 65 days. The
experiment was carried out in 250 L cone-shaped PVC tanks specially designed with a fecal decantation
column attached at the bottom where the fecal material passed into the collecting bottle. The system used
flow-through water with continuous aeration. Hatchery bred milkfish (mean body weight S. E. = 108.48
5.11g) were used to determine the digestibility of feed pea while milkfish (mean body weight S. E. = 33.27
0.87 g) were used to determine the digestibility of formulated feed peabased diets used in the growth
experiment.
For the digestibility determination of feed pea, the method by Cho et al. (1982) was adapted using a ratio of
70:30 (reference diet to test ingredients) in test diet (Table 3) while for the digestibility determination of
experimental diets, the composition of the diets were the same as in Table 1, except for the incorporation of
22
1% chromic oxide to replace an equivalent amount from the filler (Celufil). The fish were acclimated with
the control diet/reference diet (without Cr
2
O
3
) for 1 week prior to feeding them test diets containing 1%
Cr
2
O
3
. Diets were fed to satiation twice daily (0900 and 1400 h). Fecal collection was started at day 5 after
the fish were acclimated to the green diets, or when the fecal matter became greenish. The tanks and fecal
collection apparatus were cleaned twice daily, 2 hours after feeding in the morning and afternoon. Fecal
collection 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 crude
protein (AOAC, 1990) and Cr
2
O
3
(Carter et al., 1960) analyses. The test diets were likewise analyzed for
dry matter, crude protein and Cr
2
O
3
.
In vivo apparent protein digestibility (APD) and apparent dry matter digestibility (ADMD) of feed pea were
computed using the formula of Spyridakis et al. (1989) and Cho et al. (1982). ADMD and APD of
formulated diets were computed using the formula of Spyridakis et al. (1989). The data were analyzed
using ANOVA for completely randomized design (CRD). Treatment means were compared by Duncan's
Multiple Range Test (DMRT) using SAS computer software. Differences were considered significant at
P<0.05.
Results
The survival, growth, feed conversion ratio (FCR), and protein efficiency ratio (PER) data of milkfish fed
various test diets containing different levels of feed peas are summarized in Table 4. Significant differences
in both growth, survival, FCR, and PER were observed after 12 weeks of feeding. The highest weight gain,
specific growth rate, protein efficiency ratio, and survival were obtained in milkfish fed D-1, D-2 and D-3.
The best FCR were also obtained in these diets. Compared with the control (D-1), growth rate (expressed
both as % weight gain and SGR), survival, and PER decreased significantly at feed pea inclusion levels of
15% or higher, while feed conversion ratio increased. Nevertheless, it was observed that milkfish fed diets
up to 20% inclusion level with feed peas had a higher weight gain, SGR, and PER and better FCR than the
commercial feed control (D-8). Reduction in growth rate in fish fed D-4 to D-7 might have been caused by
an imbalanced amino acid content at these inclusion levels.
No differences were found in whole body composition (moisture, crude protein, crude fat, crude fiber,
nitrogen free extracts, and ash content) of milkfish fed the various test diets and control diets at the end of
the experiment (Table 5).
Values obtained for apparent dry matter digestibility (ADMD) of feed pea ingredient in milkfish was 39.0
6.0 while apparent protein digestibility (APD) was 92.2 3.5. For the experimental diets (Table 6), highest
APD value (89%) was obtained in D-3 (10% of protein replacement with feed pea). The APD values (84-
86%) for higher feed pea inclusion levels (D-5 and D-7) were not significantly different from that in D-1
(control without feed pea). No significant differences were observed in the ADMD values (6168%) of all
diets.
Over-all results show that feed peas could be used as a dietary protein source for milkfish. The optimum
level of green pea incorporation in milkfish diet is 10% of the dietary protein. This amount is about 131g
feed pea/kg dry diet. However, dietary protein levels up to about 20% feed peas (260g/kg diet) are still
acceptable.
Discussion
The feed pea meal used in the present study was produced from ground raw whole pea and complete in
terms of the typical nutritional profile of this class of feed ingredient. There was an inverse relationship
between the growth of milkfish and the dietary levels beyond 10% inclusion level of feed pea. This trend
could be the result of several factors, including reduced nutrient and energy digestibility and energy
utilization efficiency. Protein efficiency ratio also decreased and FCR increased with increasing inclusion
levels of feed pea. This may reflect a deficiency of one or more amino acids in these diets, as shown in
Table 2. It should be noted that the essential amino acid profile of the experimental diets clearly showed a
declining level in some amino acids at each feed pea increment. This was especially apparent above the
10% of the dietary protein inclusion level and may have contributed to the inferior protein utilization (PER)
and growth performance in milkfish. Perhaps the inclusion levels could be increased with amino acid
supplementation. Earlier work has confirmed that many other plant-derived ingredients may prove to be
adequate in compound feeds for fish, provided that their anti-nutritional components have been denatured
or removed. In some cases, their effectiveness may be greatly improved by supplementation with
crystalline amino acids to balance the amino acid profile of the feeds to the requirements of the fish being
fed.
23
The results of the digestibility trial did not support the assumption of reduced protein availability from feed
pea. In the present study, dietary crude protein digestibility values of the experimental diets were above
80%. A comparison with the control diet supports the observation that pea protein is well digested by
milkfish. In fact, apparent protein digestibility value was superior with the partial inclusion (10% of dietary
protein) of this plant material compared to the control without feed pea. Feed pea at this level may only
show marginal effects of anti-nutritional factors and limiting amino acids due to its relatively low content of
such compounds and its fairly good protein biological value for fish nutrition. Another consequence of using
feed 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 pea
compared to soybean meal. At this inclusion level, in practical conditions, feeding costs can be dramatically
decreased.
In general, feed pea meal proved to be an acceptable ingredient for consideration in practical diets for
grower stage production of milkfish. Future work showed be directed towards evaluating extruded pea
products where pre-gelatinization of the starch would be possible, or testing various pea protein
concentrates which have been processed to remove unwanted components such as fiber, tannin, etc.
These products contain much higher protein levels that could be used to substitute increasing amounts of
fish meal in fish feed formulations. Further studies are needed to find out the maximum inclusion level of
feed pea using other ingredient processing methods (e. g. dehulling, autoclaving, pressure cooking, protein
concentrating, etc) and different types of pellet making (e. g. extrusion); in consideration of the effect of
these processing methods on efficient utilization of essential nutrients by the fish.
Previous experiments using cold-pressed pellets for Atlantic salmon suggested that 40% fish meal protein
replacement with pea or lupin protein concentrate was feasible (Carter, 1998). Carter and Hauler (2000)
confirmed that at least 33% fish meal protein replacement with pea protein concentrate was possible. A
major problem in the use of plant meals is their relatively low protein content. In the study of Carter and
Hauler (2000), the protein concentrates used were produced by air separation and resulted in lupin and pea
concentrates with 46% and 49% crude protein, respectively. This represented an increase of between 44
and 133% over the protein in the raw lupin and peas, respectively. Protein at this level is comparable to
soybean meal, but still far lower than that in fish meal. Gouveia et al., (1993) compared the efficacy of three
legume seed meals, e. g., lupin seed meal (Lupinus albus), faba beans (V. faba) and pea seed meal (P.
sativum) in a 3-month feeding trial with juvenile rainbow trout. They reported that trout performed well on
diets that had up to 20% of the protein content being supplied by each of these ingredients. In fact, growth
and feed utilization was superior with the partial inclusion of these plant materials. The inclusion of a co-
extruded plant protein made from rapeseed and field pea had no effect at up to 15% replacement of the
protein. However, at 45% inclusion, growth performance of rainbow trout was significantly lower than the
control diet (Gomes et al., 1993). The low digestible energy from both raw and autoclaved field peas was
predicted to limit their use in rainbow trout feeds (Pfeffer et al., 1995; Gouveia and Davies, 1998). Energy
digestibility of legumes is considerably lower than for protein due to their high carbohydrate content. The
use of extrusion is important in increasing the nutrient availability of plant meals, especially in relation to
increasing the amount of digestible energy available through greater gelatinization of starch (Pongmaneerat
and Watanabe, 1993).
Since the objective of this study was to evaluate the direct nutritive value of feed peas, the experiment was
carried out in an in-door clear water flow-through system. However, experience shows that milkfish in
ponds grow better than milkfish given the same feed under controlled laboratory conditions (Sumagaysay et
al., 1991; Sumagaysay and Borlongan, 1995). It is expected that higher growth rates and better FCR and
PER may result if the milkfish are reared in ponds. It is recommended that feeding experiments using the
best diets be done under natural pond conditions and an economic assessment of the feed pea-based
feeds be conducted.
24
Table 1 Composition (g/100g diet) and proximate analyses of the experimental diets
Table 2 Amino acid composition of various test diets and a comparison of the amino acid requirements of
milkfish
Essential amino acid Diet 1 Diet 3 Diet 5 Diet 7 Milkfish Reqt*
EAA content: Calculated (g/100g diet)
Arginine 1.57 1.43 1.29 1.43 1.58
Histidine 0.68 0.60 0.54 0.46 0.60
Isoleucine 1.05 0.95 0.85 0.75 1.20
Leucine 1.79 1.64 1.48 0.77 1.53
Lysine 1.56 1.45 1.32 1.21 1.20
Methionine + Cystine 0.70 0.64 0.57 0.51 0.98
Phenylalanine + Tyrosine 1.61 1.46 1.27 1.08 1.57
Threonine 0.96 0.88 0.79 0.71 1.35
Tryptophan 0.15 0.15 0.15 0.13 0.18
Valine 1.17 1.08 1.00 0.90 1.06
EAA content: Analyzed Value (g/100g diet)
Arginine 1.21 1.61 0.94 1.30
Histidine 0.63 0.54 0.33 0.32
Isoleucine 0.96 0.98 0.64 0.74
Leucine 1.43 1.70 1.26 1.27
Lysine 1.31 1.37 1.23 0.96
Methionine** 0.27 0.31 0.2 0.22
Phenylalanine + Tyrosine 1.58 1.88 1.31 1.36
Threonine 0.92 0.82 0.72 0.64
Tryptophan nd nd nd nd
Valine 1.04 1.09 0.91 0.85
** Cystine not determined
* Based on Borlongan and Coloso, 1995
D-1 D-2 D-3 D-4 D-5 D-6 D-7 D-8
0% 5% 10% 15% 20% 25% 30% *CMF
White fish meal 10.0 10.0 10.0 10.0 10.0 10.0 10.0
Soybean meal 35.0 31.5 28.0 24.5 21.0 17.5 14.0
Copra meal 13.0 13.0 13.0 13.0 13.0 13.0 13.0
Meat and bone meal 11.0 11.0 11.0 11.0 11.0 11.0 11.0
Vitamin mix 3.0 3.0 3.0 3.0 3.0 3.0 3.0
Mineral mix 2.0 2.0 2.0 2.0 2.0 2.0 2.0
Cod liver oil 2.0 2.0 2.0 2.0 2.0 2.0 2.0
Soybean oil 2.0 2.0 2.0 2.0 2.0 2.0 2.0
Bread flour 21.2 17.3 13.9 10.6 7.2 3.8 0.4
Filler (Celufil) 0.8 1.7 2.0 2.3 2.6 3.0 3.3
Green peas - 6.5 13.1 19.6 26.2 32.7 39.3
Total 100 100 100 100 100 100 100
Crude protein 33.52 33.44 33.20 33.42 33.29 33.22 33.35 37.38
Crude fat 10.24 10.22 10.10 10.08 10.02 10.08 10.07 9.14
Crude fiber 4.66 4.96 4.42 4.39 4.94 4.51 4.63 5.20
Nitrogen-free extracts 43.54 43.34 43.96 43.37 43.33 43.79 43.70 39.18
Ash 8.04 8.04 8.32 8.74 8.42 8.40 8.25 9.10
Ingredients
Proximate compostion (%dry basis) of the test diets
*CMF = Commercial Milkfish Feed
25
Table 3 Composition (g/100g diet) of the reference and test diets for the digestibility determination
Table 4 Response of milkfish juveniles to the various test diets after 12 weeks of feeding
Table 5 Carcass proximate composition (%) of milkfish juveniles fed the various test diets
Table 6 Apparent digestibility coefficients (%) for dry matter (ADMD) and crude protein (APD) of
experimental diets in milkfish (Chanos chanos Forsskal)
Test Diet
70:30:00
White fish meal 10.0 7.0
Soybean meal 35.0 24.5
Copra meal 13.0 9.1
Meat and bone meal 11.0 7.7
Vitamin mix 3.0 2.1
Mineral mix 2.0 1.4
Cod liver oil 2.0 1.4
Soybean oil 2.0 1.4
Bread flour 20.2 13.6
Filler (Celufil) 0.8 0.8
Cr2O3 1.0 1.0
Green peas - 30.0
Total 100 100
Ingredients
Reference
Diet
SGR
(%/day)
1 (0%) 90
c
834.8
e
1.58
e
2.15
e
1.44
d
2 (5%) 90
c
833.1
e
1.51
e
2.10
e
1.46
d
3 (10%) 90
c
835.3
e
1.50
e
2.10
e
1.44
d
4 (15%) 80
b
691.0
d
1.32
d
2.25
d
1.37
c
5 (20%) 80
b
680.0
d
1.29
d
2.39
c
1.35
c
6 (25%) 70
a
512.6
b
0.89
b
2.50
b
1.21
b
7 (30%) 70
a
464.0
a
0.80
a
2.60
a
1.15
a
8(CMF) 80
b
581.4
c
1.00
c
2.46
b
1.23
b
SGR (specific growth rate) = ln (final wt.- initial wt)/84 days x 100
FCR (feed conversion ratio) = total dry feed given (g) / total wet wt. gain (g)
PER (protein efficiency ratio) = wet wt.gain (g) / amount of protein fed (g)
CMF = Commercial Milkfish Feed
PER Diet No.
%
Survival
% Weight
Gain
FCR
Diet No. Moisture
Crude
Protein
Crude
Fat
Crude
Fiber
NFE* Ash
1 (0%) 76.96 59.06 23.92 0.86 5.92 10.24
2 (5%) 75.02 60.02 23.69 0.44 4.41 10.44
3 (10%) 75.16 60.64 23.18 0.79 4.78 10.61
4 (15%) 76.24 60.46 23.16 0.58 5.56 10.24
5 (20%) 77.31 60.08 23.87 0.48 4.21 11.36
6 (25%) 76.92 59.66 22.12 0.62 6.39 11.21
7 (30%) 76.63 59.96 23.74 0.74 4.05 11.51
8(CMF) 75.00 60.56 23.71 0.44 4.25 11.04
Diet No. ADMD APD
1 (0%) 61.1
a
86.4
b
3 (10%) 68.1
a
89.2
a
5 (20%) 65.4
a
86.3
b
7 (30%) 60.1
a
84.8
b
26
Acknowledgement
The authors gratefully acknowledge the United States Department of Agriculture (USDA) through the USA
Dry Peas and Lentil Council (USADPLC), Moscow, Idaho, USA for funding support. The excellent technical
assistance of Lucia Jimenez, Marthena Joyce Bernas and Niel Tibubos is also appreciated.
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