Bangladesh]. Fish. Res., 11(2), 2007: 181-188
©BFRI
Partial replacement of fishmeal protein by soybean meal
protein in the diet of mrigal, Cirrhinus cirrhosus (Ham.) fry
D.A. Jahan*, L. Hussain and M.A. Islam
Department of Fisheries Biology and Genetics
Bangladesh Agricultural University, Mymensingh 2202, Bangladesh
*Corresponding author
Abstract
To find out a suitable balance diet for Cirrhinus. cirrhosus fry substituting fish meal
protein by full-fat soybean meal protein as dietary protein source for C. cin·hosus fry
(1.29g) was studied in 12 aquaria for 60 days. The fishes were fed with four isonitrogenous (30% protein) and iso-caloric experimental diets viz. diet 1 (100% fish meal
protein), diet 2 (75% fishmeal protein and 25% soybean meal protein), diet 3 (50%
fishmeal protein and 50% soybean meal protein) and diet 4 (25% fish meal protein and
75% soybean meal protein). Among the diets the best growth, FCR and PER were
obtained with diet 3 fed fishes and no significant difference (p<0.05) was observed
between diet 1 and 2 and diet 4 offered the worst performance. Therefore, considering
fish growth, FCR and PER, it can be suggested that fish meal protein might be replaced
50% by soybean meal protein in the diet of C. cirrhosus fry for better growth.
Key words: Fish meal, Soybean meal, C. cirrhosus fry
Introduction
Success in aquaculture depends primarily on the use of quality fish feed. Feed input is
the single largest operational cost in majority of aquaculture practices (De Silva 1985).
Fishmeal has been used traditionally as a major dietary protein source because of its
nutritive value and palatability. The scarcity of good quality fishmeal and escalating
prices thereof have generated renewed interest in the use of alternative protein sources
for fish, the need to replace fishmeal has been recognized as a major research priority
(Tacon 1993) particularly for developing countries, and thus there is a growing demand
for alternative protein sources. Consequently, plant feedstuffs with high protein content
are preferentially used in formulating diets for .most species of fish. It meets the high
protein requirement and provides an added advantage in formulations because of its
relative content of essential amino acids. The amino acid profile of soya protein is
generally superior to other plant proteins (O'keefe 2003).
Soybean meal was successfully used (without amino acid supplementation) as the
sole dietary protein sources for blue catfish (Webster et al. 1995), shrimp (Piedad-
D.A. Jahan eta!.
Pascual et al. 1990) and rainbow trout (Pongmaneerat and \'X!atanabe 1992), up to 67%
for tilapia (Shiau et al. 1987). But a few experiments were conducted on soybean meal
based feed in carp culture. It is well known that heated soybean is free from tripsin
inhibitor and to some extent methionine content of soybean feed is under the range of
carps requirement. Investigation on actual effect of soybean meal based feed on Indian
major carp like mrigal which is important culture fish in Bangladesh is important. In
view of the above, the present study was undertaken to evaluate soybean meal protein as
a partial replacement of fishmeal protein in the diet of Indian major carp, mrigal,
Cirrhinus cirrhosus fry as well as to find out a suitable balance diet for C. cirrhosus fry.
Materials and methods
The experiment was conducted in the of Department of Fisheries Biology and
Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh for 60 days in
August and September, 2003.
The experiment was conducted in 12 glass aquaria (each 44 cmx24 cmX24 em),
each containing 20-L of deep tube well water with continuous aeration. Experimental
fry were collected from Bangladesh Fisheries Research Institute, Mymensingh and
acclimatized to the laboratory conditions for 7 days prior to commencement of the
experiment and fed with a mixture of rice bran and mustard oil cake (1:1). C. cirrhosus
fry (1.29g) were randomly distributed at a rate of 10 fish per aqurium with 3 replications
for each treatment. The experimental fishes were fed twice daily at 9:00 hours and 15:00
hours with formulated diets at a fixed feeding rate of 5% body weight per day during the
entire experimental period. The ration for feeding was readjusted after weighing fish in
treatments at every 101h day on. the basis of actual average weight of the fish. Uneaten
feed was removed by siphoning 1h after each feeding. Water in each aquarium was
partially replaced twice daily during the removal of uneaten food or faeces. At the
termination of the experiment, all fishes were weighed individually and used for
analysis of whole body composition.
Roasted soybean flour was used as test ingredient. Soybean meal and fish meal were
analyzed for proximate composition. Four iso-nitrogenous diets were used in the
experiment containing 30 % protein (Akand et al. 1991), having 11% lipid where fish
meal and soybean meal were the main sources of protein. In the diets, fishmeal protein
was replaced by soybean meal protein at 0% (diet 1), 25% (diet 2), 50% (diet 3) and 75%
(diet 4), without amino acid supplementation. Diet 1 was used as a control. All the diets
were prepared in pelleted forms, using carboxymethylcellulose (CMC) as a binder. All
dry ingredients were ground finely and sieved. After sieving, all the required amount of
dry ingredients along with the vitamin premix was weighed as per formula of the
experimental diets. After well mixing of the major ingredients, soybean oil was
gradually added to the dry feed. Enough water was. added to make stiff dough. The
mixture was the passed through a hand-pelleting machine to obtain pellets and dried in
an oven at 40°C for 24 hours. The pellets were then crushed and sieved to obtain the
particle size of 500 !J.m. All the diets were packed separately in airtight plastic bags and
182
Replacement of fish meal protein by soybean meal protein
stored in a deep freeze. The formulation of feed samples from each treatment were cited
and analyzed for proximate composition and the results are presented in Table 1.
Table 1. Formulation and proximate composition of the experimental diets (% dry matter basis)
Fish meal
Soybean meal
Soybean oil
1
50.84
0.00
2.70
Diet number
2
3
38.13
25.42
17.32
34.64
01.0
01.0
4
12.71
51.96
Dextrin (starch)
CMC"
41.46
3.00
38.55
3.00
33.94
3.00
30.33
3.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
30.43 ± 0.36
10.37 ± 0.16
11.50 ± 0.21
2.20 ± 0.05
45.50 ± 0.57
18.79 ± 1.79
16.21 ± 0.24
30.36 ± 0.34
10.12 ± 0.20
9.95 ± 0.37
2.35 ± 0.06
47.22 ± 0.45
18.96 ± 6.86
16.01 ± 0.16
29.81 ± 0.28
10.26 ± 0.16
8.20 ± 0.18
2.40 ± 0.05
49.33 ± 0.61
19.25 ± 7.18
15.49 ± 0.19
30.06 ± 0.30
10.32 ± 0.12
6.11 ± 0.13
. 2.52 ± 0.10
50.99 ± 0.62
19.62 ± 0.69
15.32 ± 0.19
Feed stuffs:
Vitamin premix
Mineral premix
Proximate composition
Crude protein
Crude lipid
Ash
Fibre
Nitrogen free extract
Gross energy (kJ/g)
PE ratiob
"Carboxymethyl cellulose (Binder)
bProtein to energy ratio in mg protein I kJ of gross energy
After termination of the experiment, livers from nine fish from each treatment were
fixed in 10% neutral buffered formalin. Samples were stained with hematoxylin and
eosin and Periodic Acid-Schiff (PAS) for histological examination. Procedures were
followed according to Humason (1979).
Feed ingredients, experimental diets and fish muscle compositions (initial and
final) were analyzed for proximate composition (%) following the procedures as
described by AOAC (1990). Water quality parameters were analyzed according to APHA
(1980). All data were subjected to one-way analysis of variance CANOVA) followed by
Duncan's multiple range tests (Duncan, 1955) to test the difference between means.
Results and discussion
Results of the feeding experiment are shown in Table 2. The average final weight of
the fish increased indicating that fish belonging to different dietary groups fed actively.
No fish died during the experimental period. It indicates that feed ingredients-like
fishmeal; soybean meal has no toxic effect as soybean was roasted before feed
preparation. These results is supported by the statement of Lim and Dominy (1990) who
expressed that properly heated full-fat soybean meal destroys the antinutritional factors
and possibly increases nutritional availability, especially amino acids profile. However,
the highest attainment in fish body weight, average live weight gain and SGR were
183
D.A. Jahan et al.
recorded from diet 3 which had 50% fishmeal protein and 50% soybean meal protein
whereas those of diet 4 were significantly lower than all other diets. No significant
differences (p>0.05) in weight gain were observed between diets 1 and 2. The feed
conversion ratio (FCR) was also highest (3.12) in case of diet 4 with a tendency to
increase with increasing the proportion of soybean content in the diet. The lowest FCR
and the higher protein efficiency ratio (PER), higher apparent net protein utilization
(ANPU) were obtained in case of diet 3 and were not significantly different (p>0.05)
from those of all other dietary groups except diet 4. Relatively good (lower) FCR was
obtained by Devi et al. (1999) in case of L. rohita fingerlings fed with diet including 20
% commercially available soybean meal while feeding with 40% and 60% soybean meal
diet appreciably depressed the performance (higher FCR). In a study Akhteruzzaman
(1999) reported that 75% fishmeal protein in the diet of Indian major carp, L. rohita
fingerlings could be replaced by defatted soybean meal without any significant
reduction in growth. Davies et al. (1989) suggested that commercial soybean meal could
be replaced up to 75% offishmeal protein in diet oftilapia with no significant reduction
of average final weight. Shiau et al. (1990) also observed that defatted or full fat soybean
meal could be used to replace 30 % fishmeal protein in a diet for tilapia.
Table 2. Performance of Cirrhinus cirrhosus fry fed on experimental diets for 60 days.
Parameters
Average live weight gain(%)
Mean final body weight (g)*
SGR (%/day)
PER
FCR
ANPU (%)
Survival rate (%)
Diet number
1
160.90b
± 5.61
3.38b
± 0.09
1.60b
± 0.03
. 1.17a
± 0.04
2.86b
± 0.11
18.02a
± 0.62
100
2
164.20b
± 6.87
3.42b
± 0.10
1.63b
± 0.05
1.17a.
± 0.07
2.86b
;:!: 0.16
18.05a
± 1.05
100
3
176.703
± 3.72
3.57 3
± 0.08
1.703
± 0.03
1.22 3
± 0.05
2.73b
± 0.10
19.08 3
± 0.74
100
4
145.5c
± 2.14
3.18c
± 0.04
1.50c
± 0.02
1.07b
± 0.03
3.12 3
± 0.07
15.01 b
± 0.36
100
*Mean initial weight (g) 1.29g
*Values having the same superscripts in the same row are not significantly different (P>O.OS).
Histological studies of fish samples of liver from different treatments were
compared to the control group, which did not show any alteration from normal histology
(Fig. 1), slight irregularly arranged hepatocytes and some glycogen deposition was
observed in the liver of fish fed with the 50% soybean meal in the diet (Fig. 3), when the
level of soybean meal was used up to 75% in the diets necrotic hepatocytes with huge
empty spaces and some glycogen was present (Fig. 4). Although, the liver histology of
fish fed the 25% soybean meal in the diet was similar to the control (Fig. 2). Robaina et
184
Replacement of fish mea] protein by soybean meal protein
al. (1995) reported that areas with high levels of hepatocytes vacuolization and
disorganization were present in some samples when fish fed with 30% soybean meal
diet. While Hossain and Jaunecy (1989) observed histological abnormalities in liver
tissues of common carp fed with 50% mustard protein. The histological changes in liver
of the fishes were observed due to rearing in control condition and higher inclusion of
soybean meal protein as soybean meal also possess additional factors other than tripsine
inhibitor like phytic acid.
Fig. 1. Section of liver of Cirrhinus ci1Thosus from
Treatment 1. Normally arranged hepatocytes
with no glycogen and intercellular fat
deposition (H&Ex420).
Fig. 3. Section of liver of Cin·hinus cirrhosus
from Treatment 3. Normally arranged
hepatocytes with no glycogen and
intercellular fat deposition (H&Ex420).
Fig. 2. Section of liver of Cirrhinus cirrhosus
from Treatment 2. Normally arranged
hepatocytes with no glycogen and
intercellular fat deposition (H&Ex420).
Fig. 4. Section of liver of Ci11'hinus cirrhosus
from Treatment 4. Necrotic hepatocytes
with huge empty spaces and some glycogen
deposition are seen (H&Ex420).
The whole body compositions of the experimental fishes are shown in Table 3. It is
seen that protein and fat contents increased while moisture and ash contents decreased
after the feeding trials in all the treatments in comparison to the respective initial
values. Carcass moisture and ash content were lower with increasing amount of protein
185
D.A. Jahan et al.
and lipid. Though all the fishes were fed with iso-nitrogenous diets, the maximum
amount of carcass protein and lipid was recorded from diet 2 while the maximum
amount of ash was recorded from diet 1. Devi et al. (1999) observed higher protein and
lower lipid level in the carcass and the muscle tissues of L. rohita fingerlings fed with
soybean meal based rations compared to control (0% soybean meal). But Pongmaneerat
and Watanabe (1993) reported that using soybean meal as the sole protein source the
body protein was not greatly influenced in carps. Watanabe et al. (1992) reported that
using de-fatted soybean meal as a substitute for fishmeal in adult and juvenile yellow
tail containing 0-30% soybean meal proximate composition were not marked difference
and the lipid content was also lower in the diets containing soybean meal which is
slightly agreed with the present study. In the present study lipid content was increased
up to 25% replaced fishmeal by soybean meal but decreased gradually at inclusion of
soybean meal 50% and above.
Table 3. Whole body composition(%) of Cirrhinus cirrhosus fry fed on
experimental diets for 60 days.
Muscle
composition
(%)
Initial value
Moisture
Protein
Lipid
Ash
79.43 ± 0.41
12.32 ± 0.20
2.46 ± 0.09
3.81 ± 0.05
Final values
Diets
1
75.34 ± 0.82
13.67 ± 0.41
4.52 ± 0.43
2.76 ± 0.10
2
74.70 ± 0.70
14.07 ± 0.77
4.75 ± 0.39
2.61 ± 0.22
3
75.44 ± 0.59
13.72 ± 0.29
4.50 ± 0.36
2.57 ± 0.26
4
75.36 ± 0.73
13.64 ± 0.54
4.39 ± 0.29
2.53 ± 0.13
Results of the present study demonstrated growth performance of C. cirrhosus fry fed
on soybean meal without supplementation of amino acids. Although, the inclusion of
25% soybean meal in the diet, the growth performance of C. cirrhosus fry was
comparable to that of the control where 100% fishmeal was used but replacement of
50% fish protein by soybean protein, the growth performance of C. cirrhosus fry was
excellent. Shiau et al. (1989) observed that tilapia diet in which 100% fishmeal was
replaced by soybean meal either with or without supplementation of methionine showed
significantly lower weight gain, feed conversion ratio, protein efficiency ratio and
protein digestibility than that of the groups fed with diets containing fish meal as the
main source of protein. Akiyama (1989) reported that energy, phosphorus and
methionine levels must be considered when utilizing high levels of soybean meal in fish
feeds. Soybean meal is not nutritionally better than fishmeal but it is more cost
effective. According to Nose. (1979) the amino acid composition of soybean meal
indicates that soybean meal is deficient in methionine when compared to the amino acid
requirement of common carp fingerlings. Moreover, Viola et al. (1982) reported that
soybean meal is deficient in available energy and lysine as well as methionine, for carp
(Cyprinus carpio). So in the present study the observed reduction in growth of C. cirrhosus
fry with diet 4 might be caused by an imbalance of amino acids which was covered by
186
Replacement of fish meal protein by soybean meal protein
fishmeal protein in other diets. Therefore, results of the present study indicate that
soybean meal might be a promising ingredient as a substitute of fishmeal in the diet of
C. cirrhosus fry and SO% replacement of fishmeal might be done by soybean meal
without amino acid supplementation for better growth.
Acknowledgement
Authors are thankful to BAD-USDA-Soybean Project for providing fund to carryout the research
work. The authors are also thankful to Dr. Lutfur Rahman, Professor, Department of Genetics
and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh for kind cooperation.
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(Manuscript received 8 May 2007)
188