REPLACEMENT OF FISHMEAL WITH ALTERNATIVE PROTEIN SOURCE

FROM POULTARY WASTE MEAL IN CHANNA GACHUA JUVENILE ON

GROWTH PERFORMANCE, NUTRIENT UTILIZATION AND AMINO ACID
COMPOSITION
JAMES MILTON1*, HANIFFA M.A2KANNAN K3, RANJITH L3 KAVITHA K2
1*

College of Fishery, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction

of Ministry of Education, Huazhong Agricultural University, Wuhan, Hubei, 430070, PR

China
2

Centre for Aquaculture Research and Extension (CARE),St.Xaviers Collge

(Autonomous)Palayamkottai- 627002, Tamilnadu, India.

3

Central Marine Fisheries Research Institute (ICAR), Tuticorin Research Centre of

CMFRI,Tuticorin - 628 001, Tamilnadu, India.

Corresponding Author Email : milton.sxc@gmail.com; kannan.k84@gmail.com.

Tel : +86-13100673065 Fax : 0462 -2560670
Number of Pages :19
Number of Tables :2
Number of Figures :2

ABSTRACT
1

Three experimental diets were formulated to contain 48 g kg -1 meals, made of poultry waste
(DOP) jawala fish meal (DOJ) and anchovy fish meal (DOA). The diets were fed to juvenile
C. gachua (3.2 g) for 45 days. fish weight gain, feed intake, feed conversion ratio (FCR),
survival, specific growth rate (SGR) where found out. Feed intake, FCR and PER were
corrected for nutrients. No significant differences in feed consumption and survival were
found, but poultry waste meal diet resulted in a slightly higher final weight (14.9 for poultry
versus 13.5 and 13.2 g for DOJ and DOA), and better FCR (1.09 versus 1.2 and 1.25) and
HSI (2.0 versus 1.7 and 1.5), the differences with DOJ diet being significant (Duncan, a =
0.01). Amino acid composition for all treatments generally reflected the composition in the
muscle have twelve amino acid was found to play an important role in C. gachua. Results of
this study thus suggests that C. gachua fed diets containing poultry waste meal produce better
growth than those fed fish meal diet without showing any signs of nutrient deficiency.
Keywords: Amino acid profile, Dwarf snakehead, Channa gachua, Diet quality

INTRODUCTION

Channa gachua is commonly called dwarf snakehead, frog snakehead and brown
snakehead (Talwar and Jhingran, 1992) and the vernacular names are Para korava, Cheng,
Chenga and Koravu. It inhabits medium to large rivers, mountain streams and stagnant water
bodies. It is one among the small snakehead species and preferred both for food as well as for
ornamental purposes. Even though it is widely distributed in Asia, it has declined drastically
(vulnerable) in India (CAMP, 1998; Anjan et al., 2007) and endangered in Singapore (Lim
and Ng, 1990). Therefore, proper culture technique of this species is the prime need.
The success of intensive culture depends on the feed that contain optimum protein
prepared by locally available feed ingredients (Narejo et al., 2010). The feed thus prepared
should fulfil the need of essential amino acids, minerals, vitamins, growth promoting
substances and energy. Knowledge on specific protein requirement of fish is essential for feed
formulation and for successful intensive culture (Steffens, 1981). Some species accept
formulated feed during the last part of the yolk absorption period, i.e., after the larvae have
utilized most of their yolk reserves they start to ingest exogenous food (Balon, 1985; Kamler
1992). The optimum growth of all aquatic animals largely depends on food quality (Haniffa
and Venkatachalam, 1980; Haniffa and Arul, 1992; Joseman et al., 1994) and the quality of
food provided during the weaning of larvae is a critical step in larviculture (Devresse et al.,
1991). Qin and Fast (Qin and Fast, 1997; Haniffa et al., 2004; Haniffa et al., 2011) reported
that larval snakehead C.striatus accepted formulated feed beginning at 12 mm total length.
Qin and Fast (1996) also developed a protocol for weaning larval snakeheads (Channa
striatus) from live artemia to formulated feed.
There are several protein sources that have the potential of replacing fishmeal in
aquaculture feeds without affecting the growth performance of fish (Tacon et al., 2009). Past
research has established precedence for the partial and total replacement of fishmeal as a
protein source with poultry by-product meal, the amount of fishmeal replacement possible is
species specific (Fowler, 1991; Gouveia, 1992; Hasan et al., 1993, Hasan et al., 1997;
Steffens, 1994; Sadiku and Jauncey, 1995; Appelbaum et al., 1996; El-Sayed, 1998, Nengas
et al., 1999; Webster et al., 2000; Davis and Arnold, 2000; Sevgili, 2002). Emre et al., (2003)
studied the effect of partial replacement of fishmeal with rendered animal protein ingredients:
poultry by product meal and meat and bone meal alone and in combination with lysine and
methionine supplements in practical feed for mirror carp. Poultry waste has been widely
studied as an alternate protein source to fish meal in various fish species diets (Gallagher and
LaDouceur, 1995; Emre et al., 2003; Wang et al., 2005). As compared to fish meal, Poultry
waste meals contains a marginally lower concentration of one or more amino acids essential
3

for fish, including methionnine+cystine, lysine, and phenylalanine (Tacon and Jackson, 1985;
Davies et al., 1989; Nengas et al., 1999; Sevgili, 2002).
This study provides information on evaluation of different diets containing increasing
percentage of crude protein with essential amino acids on survival, growth performance and
FCR of murrel C. gachua.
MATERIALS AND METHODS
Fish collection and experimental set up:
The experiment was conducted in the CARE, St.Xaviers College, Palayamkottai,
Tamil Nadu, India, for a period of 60 days (2 March to 30 April 2011) during summer. It was
carried out in indoor rectangular borosilglass-reinforced glass tanks of 0.4 m 2 area each.
Water level was maintained at 50 2 cm level throughout the experimental period. Each tank
was provided with four PVC pipes of 2 cm diameter as hideouts. Water was exchanged at a
rate of 25% per 3 days.
The juvenile C. gachua were produced from induced spawning of broodstock (using
HCG) at CARE Aquafarm and they were nursery reared for 2 months in 500 L capacity FRP
tanks to attain juveniles of above 2.0 g size. The nursery-reared juvenile fish with a size range
of 3.23 3.45 g were stocked in the FRP tanks at a rate of 10 fish per tank. The treatment was
conducted in triplicates. The fishes were weighed initially and thereafter fortnightly until the
end of the experiment after removal of excess water from fishes by using nylon nets. The
samples for water quality parameters were taken at weekly intervals between 8.30 and 9.00
hours (prior to water exchange) following the standard procedures outlined by American
Public Health Association (1980). The temperature, pH, dissolved oxygen and ammoniaof the
water ranged 28.5 30.70C, 7.08.3, 4.36.5 mg L-1, 0.030.16 mg L-1respectively.
Experimental diet preparation
Poultry waste was collected from locally available shops in Palayamkottai, India. The
poultry waste was washed in water, boiled in 500C, coarsely powdered, and stored at room
temperature until used. The protein sources used in the experimental diets included high
protein soybean meal, poultry waste meal (DOP), anchovy fish meal (DOA) and jawala fish
meal (DOJ). The experimental diets were prepared separately using known quantities of
ingredients to maintain the required protein level. The selected ingredients were powdered
and sieved to get fine particles of uniform size. Then the ingredients were weighed according
to the formulation and hand kneaded by adding sufficient quantity of distilled water and
finally made into dough. The dough was then autoclaved in a closed aluminium container at
121 C for about 15 minute and then cooled. The cooled dough was separated into four parts.
4

One part was treated as control feed and the remaining parts were used for mixing poultry
waste meal, anchovy fish meal and jawala fish meal. The prepared feed were stored in air
tight containers and kept in refrigerator. Ingredients were mixed in a feed mixer and passed
through a feed extruder to make 3 mm diameter pellets. The pellets were dried in an oven
maintained at 600C. Dried feed pellets were kept in separate, tightly capped bottles and stored
in a deep freezer at 200C until used during the trial. Feed ingredients used and proximate
compositions of all the diets are presented in Table 1.
The test individuals were fed ad libitum twice a day at 10 hr and 16 hr. Water in each
aquarium was changed daily before the first feed ration was given. Unfed and dead larvae
were siphoned from each aquarium daily. Fish were sampled once in a week for growth
estimation. Five individuals from each aquarium were directly measured on a measuring
board and weight of the fish was measured using an electronic balance (sensitivity; 0.001 g).
At the end of 60th day total length and live weight of all fishes were measured and data were
tabulated.
Statistical analysis
The influence of different diets on growth parameters viz:
Survival rate = no. of final fish / no. of initial fish x 100,
Weight gain (%) = final live weight - initial live weight
SGR = (In loge final weight In loge initial weight)/ time (days)100 and
FCR = dry food consumed (g) / wet weight gain (g)
All statistical computations were performed using Duncans multiple comparisons at the
significant (P< 0.001)
Bio-Chemical and Amino Acid Analysis:
Diets were analysed for proximate composition (Table1) as follows: Protein was
determined by combustion; lipid was determined by the acid hydrolysis method; fiber was
determined by using the fritted glass crucible method; and carbohydrate (NFE) was
determined by difference. The powdered samples each 250mg were weighed accurately in an
electronic balance and transferred into labeled glass test tubes (BOROSIL). 1ml of 6M
Hydrochloric acid solution was added with sample in specified test tubes. These test tubes
were sealed at the top under vacuum by high temperature gas flame, conducted triplicates of
samples. All the sealed tubes were kept in a hot-air oven at 110 oC for 48hrs continuously.
After completion of digestion, broken the tubes at the top and transferred the digest into glass
beaker (BOROSIL), rinsed the tubes 5 times with distilled water. The acid in the digest was
evaporated to core dry under vacuum using Roto-vac evaporator. The residual content was
5

dissolved with distilled water and made-up to 6ml in a centrifuge tubes. This solution
contains 41.6g dried raw sample in 1l distilled water and used as test solution for aminoacid profile analysis by HPTLC technique. The analysis was carried out by a commercial
analysis laboratory (Manian Research Laboratory, Coimbatore, India).
RESULTS
Growth Performance:
Mean weight and growth of C. gachua fingerlings fed with 4 different kinds of feeds
over a period of 60 days are summarized in Table 2. The best average daily growth rate was
significantly higher in fish fed on poultry waste meal diet (0.064mg/g/day) followed by
control (0.053mg/g/day) whereas growth rate was poor in fish fed on jawala fish meal diet
(0.048mg/g/day). Similar trend also was observed for SGR. For instance the maximum value
was seen in poultry waste meal diet fed fish (3.51) followed by control (3.2) and the
minimum value was found in jawala fish meal fed C. gachua (3.04). poultry waste meal diet
showed the best (lowest) FCR (1.09), whereas the highest FCR value (1.25) was recorded in
anchovy fish meal diet and the difference was statistically significant (P< 0.001). The best
survival was noticed in poultry waste meal diet (76%) whereas the lowest was observed in
anchovy diet.
Biochemical Composition:
The biochemical composition of test diets is given in the Table 1. Among the four
formulated diets, the highest protein content was found in poultry waste diet (40.32%),
followed by anchovy diet (37.03%), jawala diet (35.78%) and control (25.89%) respectively.
The carbohydrate content of the C. gachua ranged between 12.05% and 13.93% in all the
diets including control diet. The lipid content varies from 5.54% (control diet) to 7.97%
(jawala diet).
Amino acid Composition:
The profile of amino acid detected through HPTLC analyses for the diets and muscle
tissue of C. gachua is represented in Figures 1 and 2 respectively. The diets showed the
presence of seventeen amino acids of which arginine, histidine, isoleucine, leucine, lysine,
methionine, phenylalanine, threonine, tyrosine and valine are essential amino acids, whereas
alanine, aspartic acid, cystine, glutamic acid, glycine, proline and serine are non-essential
amino acids.

From the results it has been detected on a whole, as the amino acid level is lower in
the control when compared to other diets viz., poultry waste meal diet, anchovy diet and
jawala diet. Among the other three diets significant difference was noticed between control
and other diets. The analyses of the muscle tissues of C. gachua revealed the presence of
twelve amino acids of which lysine, arginine, histidine, threonine, valine, isoleucine, leucine
and methionine are EAA and cystine only was the one non-EAA.
The amino acid contents viz., histidine, arginine, leucine, glycine and alanine were
found to be higher / elevated in C. gachua fed with diet poultry waste meal diet when
compared to other diets. In contrast, isoleucine content was found to be lower for the same
when compared with other diets. But as a whole as per EAAI of the muscle tissue, higher
value (246.89) has been recorded in poultry waste meal diet containing 40% of crude protein
followed by 212.46 for C.gachua fed 35% protein (anchovy diet), 208.86 for fish fed 37%
protein (jawala Diet) and the least value of 185.65 for Control diet (Figure 2).
DISCUSSION
Traditionally, Fish meal (FM) is the main and preferred source of protein used in
agricultural feed industry because of the excellent nutritional composition. However, the
rising demand and limited supply make FM an expensive protein source (FAO, 2006). The
FM price has risen from about US$ 600 per metric ton in 2005 to about US$ 2000 per metric
ton in first quarter of 2010 (IMF, 2010). The inconsistency of supply, the growing demand
and price among other problems are limiting the use of Fish meal, putting a great pressure on
the feed resource industry to find economical alternative source of protein. Alternative
protein sources such as Poultry waste (Fowler, 1991; Gouveia, 1992; Hasan et al., 1993,
Hasan and Das, 1993; Steffens, 1994; Sadiku and Jauncey, 1995; Appelbaum et al., 1996;
Nengas et al., 1999; Webster et al., 2000; Davis and Arnold, 2000), have been tested using
different fish models.
In the present study, poultry waste meal supplemented diets showed better results with
survival rate of more than 70% (P<0.01) and considerable increase in weight gain and
specific growth rate (SGR) of C. gachua. FCR (1.09) and SGR (3.51) were noticed as the
best in poultry waste meal diet. In aquaculture a feed with FCR of 1:1 and 1:2 are considered
as very efficient (Jauncey, 1982). Feed conversion ratio (FCR) values in the present study
(1.09-1.25) were similar to, previous reports of Rouse & Kahn (1998), Jones & Ruscoe
(2000), Thompson et al. (2004) and Webster et al. (2004). Poultry waste meal diet containing
40.32% crude protein was found to meet the protein requirement of C. gachua, which seems
7

to be very close to the requirement observed for H. longifilis with 45.36% (Ovie, 2003) and
hybrid H. longifilis with 45% (Olufeagba, 1999).
Wang et al. (2005) observed that poultry by product could replace 30% to 50% of FM
in Cuneate drum diet without affecting the SGR, final BW and FCR. Replacement of fish
meal with poultry by product at 80% and 100% levels in the pacific white shrimp and
sunshine bass diet respectively does not have negative effects on the FCR and BW (Emre et
al., 2003). Gallagher and LaDouceur (1995) observed that juvenile Palmetto bass fed with a
diet containing 12% fish meal and 36% low-ash poultry by product had weight gains similar
to fish fed with a diet containing 47% fish meal. Negre et al. (1999) reported that 100%
replacement of fish meal with poultry waste does not adversely affect the growth
performance of Sea breams. Poultry by products seems to be a good dietary protein
alternative to fish meals in fish diet. The observed performance indicators of C.gachua in this
study suggested that Poultry waste can comprise up to 6% of a practical diet and a complete
replacement of fish meal without compromising growth performance. Final body weight
(FBW), final body length (FBL), SGR and FCR of the C.gachua fed with poultry waste as
total replacement of fish meal were similar to those fed with control diets. These results are in
agreement with Webster et al. (1999); who reported that Sunshine bass fed with a diet
containing poultry by products and soy bean meal as a complete replacement of fish meal had
similar growth performance as fish fed diets containing fish meal. The FCR and PER values
observed in this study were within the range reported by Soltan (2009) who replaced dietary
fish meal with poultry by product plus different grain sources and enzyme supplements in
Nile tilapia diets.
Amino acids are the building blocks of proteins and serve as body builders. They are
utilized to form various cell structures, of which they are key components and they serve as
source of energy (Babsky et al., 1989). The results of the present study indicated that dwarf
snakehead fed with poultry waste meal diet (40% protein) replacing the fish meal is
suggestive for the betterment of survival and growth of C. gachua. This study is in
accordance with the evaluation of practical diets level with / without fish meal for juvenile
red claw fish (Webster et al., 2004), where 35% protein was found to be essential for their
growth. Our study is also supported by Muzinic et al. (Muzinic et al., 2004) where juvenile
red claw fed with 40% protein showed significant growth rate.
The gross dietary protein requirement is influenced directly by the amino acid
composition of the diet (Wilson, 2002). Carbohydrate provides immediate energy and
increases protein sparing effect on growth (Shiau and Peng, 1992). Total lipid including
8

cholesterol is also essential for survival and growth of crustacean (Mullen and Martin, 1992).
It has been reported that the amino acids viz: arginine, alanine and glycine stimulated
ingestion in sea bream larvae (Kolkovski et al., 1997). Tryptophan plays an important role in
the brain as a precursor of the neurotransmitter; serotonin which has a major effect on the
feeding behaviour of animals (Kolkovski et al., 1997). Histidine is involved in many
metabolic functions including production of histamines, which take part in allergic and
inflammatory reactions. It plays an important role in maintaining the osmoregulation and
energy production (Abe and Ohmama, 1987). Methionine and lysine are generally critical in
feed formulation (Takon, 1993). Glycine, which is one of the major components of human
skin collagen, together with other essential amino acids such as alanine, proline, arginine,
serine, isoleucine and phenyl alanine form a polypeptide that will promote regrowth and
tissue healing (Kanazawa et al., 1977; Heimann, 1982; Witte et al., 2002).
The amino acid profile of the practical diets has also been done revealing the presence
of essential amino acids, viz., Phenylalanine, threonine, tyrosine, methionine etc, in higher
amount in poultry waste meal diet with 40% protein than other diets. This suggests that the
specific functions governed by these amino acids were performed well in the fry fed with
poultry waste mixed diet. Replacement of fish meal with poultry waste meal appears to
provide sufficient levels of essential amino acids to meet the necessary requirements of
C.gachua. Webster et al. (Webster et al., 1994) indicated that juvenile red claw (Small
0.025g) required 33% dietary protein to achieve highest / maximum growth rate; whereas
Manomaitis (Manomaitis, 2001) evaluated protein requirements of two age classes of
juvenile red claw and reported that small (0.02g) juveniles required 40% dietary protein
whereas diet with 30% protein for large (3.0g) red claw. Thus instead of taking anchovy and
jawala (where significantly lower protein content observed), poultry waste meal with protein
content of 40% is proved to be effective as per survival and growth and also essential amino
acids index (EAAI).Elimination of fish meal as corresponding substitute with poultry waste
meal showed modest change in EAAI, that is significant increase was observed in C.gachua
fed with Control (with no added fish meal). This proves the importance of essential amino
acids for the growth of fish.
However, we observed that fish consuming the diet containing poultry waste had a
significantly lower PER than those fed the diet containing fish meal (1.95 vs 2.13, P<.01).
The lower PER value resulting from the PBM diet could be due to the: i) limiting amino acid
(histidine, methionnine+cystine, lysine and phenylalanine) content (Tacon and Jackson, 1985;
Nengas et al., 1999; Sevgili, 2002), High processing temperature of poultry wastes may
9

reduce digestibility of protein and amino acids due to the lysine and cystine+cystein losses
(Opstvedt et al., 1984; McCallum and Higgs, 1989). Nontheless, the results of this study
confirm the conclusions from previous studies that poultry wastes could be used as
alternative to fish meal at various levels without adversely affecting the growth performance
of fish: 20% in feeds for Chinook Salmon (Fowler, 1991), 25% for silver Sea bream (ElSayed, 1994), 21% for Australian Snapper (Quartararo et al., 1998), 71% for gilthead Sea
bream (Nengas et al., 1999), and 14% for red drum (Kureshy et al., 2000) and sunshine bass
(Webster et al., 2000). Based on previous studies and our findings, poultry waste is a good
alternative to fish meal in murrel diets and poultry waste could be used to replace fish meal in
aquaculture feed without affecting growth performance.
In the present study, it is suggested that poultry waste meal is a better supplementary
material than fish meal since it has produced better growth in C. gachua. Moreover anchovy
mixed diet also produced favourable results regarding growth of C. gachua when compared
to jawala diet. But in the view of selecting a profitable supplement in feed management
practices, use of poultry waste meal may be recommended since it is a biowaste polluting the
environment. Hence it is clear that poultry waste meal can be recommended as an alternative
ingredient for fish meal in feed formulation of freshwater murrel culture as well as fish and
shrimp culture.
ACKNOWLEDGEMENTS
The work was supported by CSIR Emeritus Research Grant (No.21 (0670)/07/EMRII) sanctioned to Dr. M. A. Haniffa. We are grateful to Rev. Dr. Alphonse Manickam S.J.,
Principal, St. Xaviers College, Palayamkottai for providing necessary facilities.

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15

Table 1. Ingredient used and proximate composition of the experimental diets (g kg-1 DM)

DOP

DOA

DOJ

Ingredients

Control

Soy Flour

25

10

10

10

Tapioca Flour

20

10

10

10

Wheat Flour

20

10

10

10

Rice Flour

20

10

10

10

Rice Bran

10.7

7.5

7.5

7.5

Fish Oil

2.3

2.3

2.3

2.3

Vitamin / Mineral Mix *

Anchovy (%)

48

Jawala (%)

48

Poultry waste (%)

48

Crude Protein (%)a

25.89

40.32

37.03

35.78

Crude Carbohydrate (%)a

12.05

12.25

13.93

12.23

Crude Fibre (%)a

2.22

2.00

2.38

2.10

Crude Lipid (%)a

6.12

5.54

6.54

7.97

Crude Ash (%)a

15.04

14.45

19.02

17.05

Proximate composition, g kg-1

* Vitamin-mineral premix (mg Kg-1 diet): 80:riboflavin, 6.0:biotin, (IU) vitamin A, 6000:vitamin ANCHOVY
DIET, 2000:vitamin E, 6000 IU, 0.1:nicotinic acid, pantothenic acid, 180 menadione, 0.6: thiamin
hydrochloride, 15:pyridoxine, 40:inostol, 400:astaxanthin, 60:choline chloride, Zinc (as sulphate)72, iron (as
sulphate) 36, manganese (as sulphate) 12, selenium (as selenate) 0.2, chromium (trivalent, as chloride) 0.8,
iodine (as iodate)1.2, copper (as sulphate) 24, cobalt (as chloride) 0.6 and molybdenum (as molybdate) 0.2.

16

Table 2. Survival and growth of fry of Channa gachua (Hamilton) fed on different feeds.
Parameters

Control

DOP

DOA

DOJ

Initial Weight (g)

3.23 0.24

3.07 0.15

3.26 0.20

3.37 0.27

Final Weight (g)

13.6 0.30

14.9 0.18

13.5 0.24

13.2 0.44

Weight gain (g)

10.37

11.83

10.24

9.83

Growth rate (mg/g/day)

0.053

0.064

0.052

0.048

FCR

1.230.12

1.09 0.11*

1.25 0.12*

1.20 0.15

SGR (%)

3.20.13

3.51 0.09*

3.14 0.12

3.040.17*

72

76*

70

75

Survival

(%)

Each value is mean SD of 10 individual. *Significant at 1.0% level (P < 0.01).

17

Figure1 Percentage of essential Amino acids composition (g/ 100 g wet wt.) of different
diets

18

Figure 2. Percentage of essential Amino acids composition (g/ 100 g wet wt.) of muscle tissue
of fry C.gachua

19