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Citation of this paper

Effect of replacement of fishmeal with processed soybean on growth performance and nutrient retention in Channa punctatus (bloch.) fingerlings

M Jindal, S K Garg, N K Yadava and R K Gupta

Department of Zoology and Aquaculture, CCS Haryana Agricultural University, Hisar 125 004 (Haryana)
mypshya@rediffmail.com

Abstract

To study the growth performance of fish Channa punctatus, ten experimental diets (D1 to D10) were formulated by replacing fishmeal (FM) with hydrothermically processed soybean (HPS) at four inclusion levels (25, 50, 75 and 100%) from the control diet with and without supplementing the diets with mineral premix and amino-acids (MPA).

 

Studies have revealed an enhancement in the live weight gain, growth per cent gain in body weight, specific growth rate, apparent protein digestibility, protein efficiency ratio and energy retention in the fingerlings fed on diets containing HPS at high (75-100 gmg-1 of diet)  inclusion levels. Crude protein, fat and energy contents in the fish body also increased, while those of moisture and ash decreased significantly (P<0.05) in the fish fed on diets where most of the FM was replaced with HPS. An analysis of the water samples collected from the treated waters revealed that total ammonia and reactive phosphate excretion decreased with increase in the inclusion levels of HPS in the diets depicting a negative correlation with growth and digestibility and positive with feed conversion ratio. Such a replacement could save total feed costs and alleviate pollution problems in intensive aqua-cultural systems.

Keywords: Carcass, Channa punctatus, excretion, fishmeal, growth, protein digestibility, processed soybean, replacement


Introduction

Fish feeding represents over 50% of operating costs in intensive aquaculture, with protein being the most expensive dietary source. The ultimate aim of artificial feeding in aquaculture is to achieve maximum protein deposition with minimum inputs of feeds at a minimum cost. The development of commercial aqua feeds has been traditionally based on fishmeal (FM) as the main protein source due to its high protein content and balanced essential amino acids (EAA), digestible energy sources, minerals and vitamins. FM is the most expensive protein source in animal and aquaculture feeds (Tacon 1993). The increasing costs and unpredictable availability of FM necessitates the search for its replacement with cheaply and abundantly available plant protein feed stuffs such as soybean, rapeseed meal, guar, moong, sorghum etc. due to their low prices and availability (Robinson and Menghe 2007). Due to worldwide dominance of soybean and its appreciation as quality protein, several workers have attempted to replace FM with soybean meal (SBM) in diets for several fish species (Watanabe and Pongnaneerat 1993, Kaushik et al 1995, Degani 1987, Webster et al 1995, Boonyaratpalin et al 1998, Garg 1998, 1999, Nyirenda et al 2000, Kalla et al 2003). Studies have revealed that the use of SBM supplement in fish feed not only reduces the cost of feed formulation but also reduces the postprandial excretion of total ammonia and reactive phosphate in the water (Kim et al 1995a, Vielman et al 2000).

 

Presence of endogenous anti-nutrient factors (ANF’s) i.e. lectin, phyto-haemoglutinin, anti vitamins and protease (trypsin) inhibitors (Liener 1980), however, limits their use. Development of new processing technologies for the removal of ANF’s has facilitated their increased use as protein source in the fish feed (Garg et al 2002). Therefore, for the elimination of ANF’s, soybean seeds (Glycine max) were hydrothermically processed before incorporating in the diets. Present studies examine the effects of hydrothermically processed soybean (HPS-Plant protein) and FM (animal protein) in the fingerlings of Channa punctatus on their growth, nutrient utilization and body composition. Effect of protein sources on postprandial excretion of total ammonia and reactive phosphate in the treated waters were also examined.
 

 

Materials and methods

 

Experimental diets

 

A control diet was prepared using FM (as the chief protein source), groundnut oil cake and rice bran. Processed full fat soybean was used as FM replacer and prior to incorporation it was subjected to hydrothermical treatment at 15 lbs at 1210C for 15 minutes for the removal of ANF’s (Garg et al 2002). Ten isocaloric and isonitrogenous diets (D1 to D10) were formulated according to Table 1. Diets D1 and D6 were used as control diets. The proximate compositions of the formulated diets are also given in Table 1.


Table 1.  Ingredient and Proximate Composition (% dry weight basis) of experimental diets (D1 to D10)

Ingredients

Diet Number

D1

D2

D3

D4

D5

D6

D7

D8

D9

D10

Groundnut Oil Cake

60.0

60.0

60.0

60.0

60.0

60.0

60.0

60.0

60.0

60.0

Rice Bran

24.0

24.0

24.0

24.0

24.0

23.0

23.0

23.0

23.0

23.0

Fish Meal (FM)

10.0

7.5

5.0

2.5

-

10.0

7.5

5.0

2.5

-

Hydrothermically treated Soybean

-

2.5

5.0

7.5

10.0

-

2.5

5.0

7.5

10.0

Chromic Oxide ( Cr2O3)*

1.0

1.0

1.0

1.0

1.0

1.0

1.0

1.0

1.0

1.0

Binder (Carboxyl methyl cellulose)

5.0

5.0

5.0

5.0

5.0

5.0

5.0

5.0

5.0

5.0

MPA**

-

-

-

-

-

1.0

1.0

1.0

1.0

1.0

Proximate Compositionm %

Crude Protein

31.5

30.9

30.6

29.3

29.5

31.8

31.2

31.6

31.3

30.9

Crude Fat

8.9

9.2

9.4

9.5

9.7

8.9

9.2

9.3

9.6

9.7

Crude Fiber

9.3

9.5

9.5

9.4

9.4

9.3

9.5

9.4

9.4

9.4

Total Ash

11.4

10.9

10.3

9.5

8.8

11.9

11.2

10.3

9.7

9.1

NFE

38.9

39.6

40.3

40.2

42.7

38.1

39.0

39.3

40.0

42.7

Gross Energy, KJg-1

17.6

17.7

17.9

18.0

18.1

17.6

17.7

17.8

18.0

18.1

* - Cr2O3 was used for estimating apparent digestibility

** - mineral premix and amino acid (MPA) supplemented @ 10gkg-1 diet

(each Kg contains Cu – 312 mg; Co – 45 mg; Mg – 2.114 g; Fe – 979 mg; Zn – 2.13 g;           I – 156 mg; DL-Methionine – 1.92g; L-lysine mono hydrochloride – 4.4 g; Ca – 30%;             P – 8.25%)

Diets D1 to D5 not supplemented with MPA; Diets D6 to D10 supplemented with MPA.

Diets D1 and D6 were used as control diet.


Experimental design

 

Fingerlings  of Channa punctatus (Mean body weight 12.93+ 0.70) were obtained from fish dealers of Hisar during April-May and acclimated in the laboratory for a minimum period of 7 days and were fed ad libitum on a feed containing FM as the protein source (D1) between 0800 – 0900 h.

 

All treatments were conducted in transparent glass aquaria (60 30 30 cm), containing 30L of chlorine free water, in replicate of three, kept in the laboratory where the temperature was maintained at 2010 C and a lighting schedule at 12:12h of light. All groups of fish were fed daily only once between 0800 – 0900 h, at the fixed feeding rate of 2% BWd-1 for the whole experimental duration of 45 days on different diets depending upon the treatment.

 

Fish were bulk weighed every 10th day and feeding rates adjusted accordingly. In order to maintain water quality, water in the aquaria was replenished daily with the water which had been previously equilibrated to the desired temperature (200C).

 

Fish fry were fed with respective diets and thereafter, the uneaten feed was siphoned out and stored separately for calculating the feed conversion ratio (FCR). Faecal matter was collected by pippeting following the method of Spyridakis et al (1989) every morning. The pooled faecal samples were dried in an oven maintained at 600C for subsequent analysis. Individual weight of the fish fry was recorded at the beginning and at the end of the experiment.

 

Analytical techniques

 

The feed ingredients, experimental diets, faecal samples and fish carcass (initial and final) were analysed following the procedure of AOAC (1995). Chromide oxide levels in the diets as well as in the faecal samples were estimated spectrophotometrically following the method of Furukawa and Tsukhara (1966).

 

Live weight gain (g), growth percent gain, specific growth rate (SGR, %d-1), protein efficiency ratio (PER) and gross conversion efficiency (GCE) were calculated using standard methods (Steffens 1989). Apparent nutrient digestibility (APD) of the diets were calculated according to Cho et al (1982) as follows:


At the end of feeding trials, water samples from each aquarium were collected at 2h interval for the estimation of excretory levels of total ammonia (NH3-N) and reactive phosphate (P) following APHA (1998) and calculated as follows:

 

 

ANOVA, followed by Duncan’s multiple range test (Duncan 1955) was applied to find out the significant differences between different treatments. Correlation coefficient was determined between different variables.

 

Results

 

The proximate composition of the diets revealed that crude protein ranged between 29.34 – 31.50%, crude fat 8.86 – 9.70%, crude fiber 9.3 – 9.5%, total ash 9.06 – 11.93% and energy 17.57 – 18.12 KJg-1 (Table 1).

 

Growth and nutrient retention

 

Mortality was low and independent of the experimental treatments (Table 2).


Table 2. Growth performance of fish Channa punctatus fed on ten experimental diets containing fish meal and processed soybean meal at various inclusion levels

Diet No.

Weight gain, g

Growth,

% gain

Growth/ day, %

SGR

FCR

APD

GCE

PER

NH3-N

Reactive P

Mortality

%

mg100g-1 BW of fish

D1

4.473A

34.052A

0.640A

0.651A

2.868A

75.953A

0.348A

0.142A

0.606

0.276

20

S.E.

+ 0.049

+ 0.299

+ 0.004

+ 0.004

+ 0.032

+ 0.339

+ 0.004

+ 0.001

+0.003

+0.006

D2

4.690AB

35.443A

0.669AB

0.674A

2.771AB

76.963B

0.361A

0.151AB

0.553

0.263

10

S.E.

+ 0.043

+ 0.362

+ 0.005

+ 0.005

+ 0.026

+ 0.057

+ 0.003

+ 0.001

+0.003

+0.008

D3

5.023C

38.694B

0.720C

0.726C

2.597C

78.457C

0.384B

0.164C

0.523

0.253

-

S.E.

+ 0.006

+ 0.270

+ 0.004

+ 0.004

+ 0.003

+ 0.264

+ 0.000

+ 0.000

+0.008

+0.008

D4

5.223CD

40.398BC

0.747CD

0.754CD

2.518CD

82.143D

0.397B

0.173CD

0.496

0.226

20

S.E.

+ 0.033

+ 0.322

+ 0.005

+ 0.005

+ 0.016

+ 0.380

+ 0.002

+ 0.001

+0.003

+0.008

D5

5.360DE

41.197CD

0.759DE

0.766DE

2.476CE

85.297E

0.404BC

0.182DE

0.473

0.190

-

S.E.

+ 0.105

+ 0.717

+ 0.011

+ 0.011

+ 0.047

+ 0.104

+ 0.007

+ 0.005

+0.003

+0.005

D6

4.583AF

33.839A

0.642A

0.647A

2.844AF

76.827B

0.352A

0.144A

0.506

0.240

10

S.E.

+ 0.171

+ 1.083

+ 0.017

+ 0.018

+ 0.103

+ 0.088

+ 0.013

+ 0.004

+0.003

+0.000

D7

4.850BCF

35.847A

0.675B

0.680 A

2.703BCF

79.137C

0.370AB

0.155BC

0.490

0.216

20

S.E.

+ 0.043

+ 0.327

+ 0.005

+ 0.005

+ 0.024

+ 0.426

+ 0.003

+ 0.001

+0.000

+0.003

D8

5.193CE

39.980 BD

0.740CE

0.747CE

2.551C

80.823F

0.392B

0.169C

0.480

0.203

20

S.E.

+ 0.013

+ 0.089

+ 0.001

+ 0.001

+ 0.006

+ 0.160

+ 0.001

+ 0.000

+0.005

+0.003

D9

5.676G

43.797E

0.798F

0.807F

2.381DEG

83.517G

0.420CD

0.187E

0.453

0.190

-

S.E.

+ 0.017

+ 1.314

+ 0.019

+ 0.020

+ 0.075

+ 0.280

+ 0.013

+ 0.007

+0.003

+0.000

D10

5.983H

45.883F

0.829G

0.839G

2.282G

87.990H

0.438D

0.199F

0.426

0.173

10

S.E.

+ 0.113

+ 0.815

+ 0.011

+ 0.012

+ 0.042

+ 0.324

+ 0.008

+ 0.005

+0.008

+0.003

All values are mean + S.E. of mean  

Mean with same letter in the same column are not significantly (p>0.05) different.

Data were analysed by Duncan’s Multiple Range Test


Growth, per cent gain in body weight and SGR (%) increased with each increase in the inclusion levels of HPS up to 100 gKg-1 of diet in comparison with the fish fed on diet containing FM as the major protein source (Table 2, Figure 1).


D1  to D5  : Without MPA;    D6  to D10  : With MPA


Figure 1.  
Weight gain (g) in Channa punctatus fed on diets D1 to D10 containing fishmeal and processed soybean (without and with MPA) at various inclusion levels.


Studies have revealed that FCR and APD values improved with each increase in the inclusion of HPS. A further improvement in growth and digestibility took place on supplementing the diets with mineral premix and amino acids (MPA) (Figure 1). Significantly (P<0.05) highest values, thus, were observed in fry fed on diet D10 containing 100g of soybean Kg-1 of diet and supplemented with MPA. Similar effect of FM replacement with HPS were observed in PER, SGR and GCE.

 

Proximate carcass composition

 

Protein, fat and energy contents increased, while those of moisture and ash decreased with each increase in the inclusion level of HPS. Significantly (P<0.05) highest protein, fat and energy levels were seen in fish fed on diet D10 in comparison with fish fed on diet D1 containing FM as the major protein source (Table 3).


Table 3.  Proximate Carcass Composition ( % fresh weight ) of the fish Channa punctatus fed on ten experimental diets from D1 to D10

Diet No.

Moisture

Crude Protein

Crude fat

 

Total Ash

NFE

Energy, KJg-1

D1

77.930A

12.500A

3.756A

3.110AE

2.710A

4.905A

S.E.

+ 0.322

+ 0.232

+ 0.061

+ 0.040

+ 0.040

+ 0.076

D2

77.287AB

13.247B

3.950B

3.060AB

2.463B

5.115B

S.E.

+ 0.397

+ 0.250

+ 0.101

+ 0.047

+ 0.029

+ 0.098

D3

76.227C

14.137C

4.173C

3.056AC

2.406BC

5.405C

S.E.

+ 0.274

+ 0.168

+ 0.057

+ 0.033

+ 0.032

+ 0.066

D4

76.073C

14.377C

4.226C

2.966D

2.366BD

5.475C

S.E.

+ 0.101

+ 0.139

+ 0.023

+ 0.026

+ 0.084

+ 0.025

D5

76.000C

14.357C

4.296C

2.960D

2.393BE

5.503C

S.E.

+ 0.534

+ 0.378

+ 0.103

+ 0.057

+ 0.073

+ 0.129

D6

76.840BC

13.303B

3.990BD

3.163AE

2.706A

5.185BD

S.E.

+ 0.015

+ 0.066

+ 0.020

+ 0.013

+ 0.800

+ 0.005

D7

76.547C

13.957C

4.133CD

3.120AE

2.260CDEF

5.319CD

S.E.

+ 0.012

+ 0.013

+ 0.026

+ 0.011

+ 0.011

+ 0.011

D8

76.380C

14.110C

4.186C

3.000BCD

2.330BE

5.391C

S.E.

+ 0.010

+ 0.080

+ 0.018

+ 0.000

+ 0.090

+ 0.008

D9

75.960C

14.930D

4.280C

2.980BCD

1.850G

5.539C

S.E.

+ 0.043

+ 0.111

+ 0.005

+ 0.017

+ 0.112

+ 0.009

D10

75.070D

15.710E

4.500B

2.986BCD

1.750G

5.792E

S.E.

+ 0.034

+ 0.023

+ 0.015

+ 0.003

+ 0.000

+ 0.011

All values are mean + S.E. of mean;

Mean with same letter in the same  column are not significantly (p>0.05) different.

Data were analysed by Duncan's Multiple Range Test


Postprandial excretory levels of total ammonia (nh3-n) and reactive phosphate (p)

 

 Postprandial excretory levels of NH3-N and reactive P. were found high in the treated waters for the fish fed on experimental diets containing FM as the protein source. The excretion decreased on increasing the inclusion levels of HPS. Peak values in NH3-N excretion occurred after 6-8h and that of reactive P occurred after 8h of feed distribution. The peak time of excretion of NH3-N and reactive P for the groups of fish fed on experimental diets containing HPS at varying inclusion levels was slightly earlier than the groups of fish fed on FM as the only source of protein. A further reduction in the excretion of NH3-N and reactive P was observed when diets were supplemented with MPA indicating that incorporation of MPA reduces the excretion of NH3-N and reactive P in the water medium (Figure 2 and 3).


A

B

 

Figure 2.  Excretion patterns of total ammonia (mg/100g Body weight, BW) in treated waters in fish Channa punctatus fed on diets D1 to D10 "A" without MPA (D1to D5) "B" with MPA (D6  to   D10 )



A

B

 

Figure 3.  Excretion patterns of Ortho-phosphate (mg/100g Body weight, BW) in treated waters in fish Channa punctatus fed on diets D1 to D10 "A" without MPA (D1to D5 ) "B" with MPA (D6  to   D10 )


Discussion

 

In the present studies, significantly (P<0.05) higher growth and digestibility was observed in Channa punctatus fed on diets containing HPS which indicate that partial/complete replacement of FM was possible with the incorporation of processed full fat soybean and also on supplementing the diets with MPA without compromising the growth performance, digestibility and nutrient retention in Channa  punctatus.

 

Many authors have reported depression in growth in fish fed on soybean containing diets. The poor growth obtained in tilapia on replacing FM by raw soybean (Mazid et al 1994) was attributed to the presence of ANFs present in the untreated soybean. Raw legume seeds contain several ANFs (Liener 1980), which may be destroyed by thermal treatment (Garg et al 2002). Huisman and Van der Poel (1991) concluded that trypsin inhibitors can be eliminated by atmospheric steaming (1020C) for long processing times. However, processing time which exceeds 40 minutes do not further increase apparent digestibility for nitrogen (Garg et al 2002).

 

The apparent protein digestibility (APD) and energy retention increased with increase in the inclusion levels of plant proteins in the diets. These results are similar to those observed on a cyprinid Labeo rohita for soybean (Hossain et al 1997). Studies have further revealed that APD had no effect on protein and energy retention of the fish (Bureau et al 2000). In general, the pattern of APD corresponds to growth trends of fish fed on different diets incorporating plant origin proteins (Table 2).The high protein digestibility of plant origin feeds in comparison to FM based feeds may be related to the omnivorous/herbivorous feeding habits of Channa  punctatus. These findings strengthen the view that the enzyme system in Channa punctatus are better equipped to digest and assimilate nutrients from plant origin feeds.

 

Present studies have revealed that growth and digestibility in Channa punctatus was further enhanced when HPS diets were supplemented with MPA. Significantly (P<0.05) higher growth was observed in fish fed on diet D10. The results are in agreement with Webster et al (1995), who have succeeded in achieving complete replacement of FM by the use of HPS in the catfish, Ictalurus furcatus with supplement of methionine (Robinson and Menghe 2007) in catfishes. Supplement of MPA was also found to enhance the growth of eels. Garg (1998, 1999) was also able to achieve higher growth coupled with low FCR in Cirrhinus mrigala fingerlings fed on HPS based diets supplemented with MPA and observed enhanced growth, high digestibility and high retention of protein and phosphorous.

 

Studies in which SBM was successfully used as partial or complete replacer for FM were reported for Tilapia (Mazid et al 1994), Labeo rohita (Khan and Jafri 1994), blue catfish, Ictalurus furcatus (Webster et al 1995), common carp, Cyprinus carpio (Kim et al  1995 a, b), Cirrhinus mrigala (Garg 1998, 1999),  Mugil cephalus (Kalla et al 2003).

 

With the incorporation of HPS in the diets crude protein, fat and energy values increased, while those of moisture and ash decreased. These results appear to be in agreement to those of Ray and Das (1992) on Labeo rohita and Shearer (1994) on trout and Kalla et al 2003 on Mugil cephalus.

 

Fishes excrete nitrogen mainly as ammonia which is found to be influenced by dietary protein quality and quantity (Kaushik and Cowey 1991). The reduction in NH3-N and reactive P excretion was found to be negatively correlated with the growth and digestibility parameters in Channa punctatus and a positive correlation was observed with FCR values.

 

According to Viola and Lahav (1993) calculated amounts of excreted (not retained) nitrogen per Kg gain was reduced by 20% with the lysine supplemented feeds, as compared too the 30% protein feed. Concomitantly, calculated phosphorous excretion per Kg gain was also decreased approximately by 100%.

 

The decrease in NH3-N and reactive P excretion in the treated water with the use of proteins of plant origin in feed has important implications on the management of highly intensive farming system. Therefore, more studies are warranted in this area.


Conclusions

 

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Received 17 June 2007; Accepted 26 September 2007; Published 1 November 2007

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