Aquaculture

ELSEVIER Aquaculture 156 (1997) 241-249

Interactions of dietary levels of protein and energy

on fingerling snakehead, Channa striate
K. Samantaray *, S.S. Mohanty
Fish Nutrition Laboratory, College of Fisheries. Orissa U&x. of Agri. and Tech., Rangailunda, Berhampur
760 007, Ganjam, Orissa, India

Received 10 October 1996; revised 5 February 1997; accepted 5 May 1997

Abstract

A 4 X 3 factorial experiment was conducted to determine the optimum protein to energy

(P/E) ratio for fingerling snakehead. Four crude protein levels and three energy levels at each
protein level were utilized. Laboratory-made brown fish meal was used as the major source of
protein in all the test diets. Each of the experimental diets was fed to triplicate groups of 15
fingerlings with an average individual weight of 12 g in 1IO-I flow-through fibre blast tanks at
28 f 2°C with 5.9 to 7.2 mg dissolved oxygen I-‘. The diets were fed at a rate equalling 3 wet
body wt% day ’ in two equal rations and adjusted weekly for g-weeks. The highest growth was
attained by fingerlings fed a 40% protein diet with a P/E ratio of 90.9 mg protein kcal-’
digestible energy. A lower but not significantly different (P > 0.05) growth response was attained
by fingerling fed a 45% diet with a P/E ratio of 93.8 mg kcal- ‘. Protein efficiency ratio (PER)
was affected by the increase in energy level from 400 to 480 kcal 100 g-’ at all protein levels
except 40% where a diet with energy 440 kcal 100 g- ’ showed maximum PER value. The lowest
feed conversion ratio (FCR) was obtained at an P/E ratio of 90.9 mg protein kcal _ ‘. Carcass
moisture content was not affected by dietary P/E ratio. Other changes in carcass composition
due to the change in dietary P/E ratio were not consistent. This study also indicated that a lipid
level of 13% can be effectively utilized by snakehead fingerlings without any adverse effects.
0 1997 Elsevier Science B.V.

Keyvords: Channa striata; Snakehead; Murrel; Protein requirement; Protein to energy ratio

1. Introduction

Channa striata, snakehead or murrel, is a cultivable food species in the Indian

subcontinent. Its air-breathing characteristics and general hardiness allow them to be

* Corresponding author. Tel.: f91 680 282235: fax: +91 674 407780.

00448486/97/$17.00 0 1997 Elsevier Science B.V. All rights reserved

PII SOO44-8486(97)00140-3
242 K. Samantaray, S.S. Mohanty/Aquaculture 156 (19971241-249

cultured in areas which are not suitable for the culture of Indian major carps. Previous
studies have shown that snakehead fingerlings require about 50% crude protein for
maximum growth (Wee and Tacon, 1982; Wee, 1986) whereas the fry require about
55% crude protein for maximum growth (Mohanty and Samantaray, 1996). The protein
requirement of any fish varies with water temperature, water quality and fish size
(National Research Council, 1977). The non-protein energy level may also influence the
protein requirement of fish. Prather and Love11 (1973) postulated that high levels of
protein without sufficient energy in the diet may be harmful to fish. Page and Andrews
(1973) observed that larger size fish require more energy and less protein compared to
smaller fish.
The protein sparing effect of fat and carbohydrate has been studied in other fishes
(Cho and Kaushik, 1990). The protein to energy (P/E) relationships for a number of
fishes, maintained under various culture conditions, have been reported. Mukhopadhyay
et al. (1986) observed that a P/E ratio of 87.6 mg protein kcal- ’ produced maximum
growth in Clarias batrachus, whereas 95 mg protein kcall’ was found to be optimal for
Labeo rohita (Das et al., 1991). For Indian carps, an P/E ratio of 8.4 and 10.6 mg
protein kcal-’ for fingerling and advanced fingerling, respectively, have been reported
(Seenappa, 1992).
The present study was conducted to evaluate the response of murrel fingerlings to
diets containing various protein levels and P/E ratios and to determine the maximum
inclusion level of dietary lipid to spare protein for growth.

2. Material and methods

2.1. Diets

12 experimental diets were formulated using laboratory-made brown fish meal as the
major source of protein to have three digestible energy levels (400, 440 and 480 kcal) at
4 protein levels (35, 40, 45 and 50%) (Table 1). The energy level at each protein level
was carefully maintained in the present study to determine the optimum P/E ratio for
each energy level. Lipid constituted 9, 13 and 17% of the dry diet at each protein level.
Since the energy values for the dietary ingredients utilized in the study are not available
for snakehead, the energy value of each diet was calculated based on standard physio-
logical fuel values of 4,4 and 9 kcal g- ’ for protein, carbohydrate and lipid, respectively
(Pike and Brown, 1967). Inevitably, there were slight alterations in the percentage
contribution of ingredients between the diets primarily those of cellulose and rice bran.
The energy levels were adjusted by varying the quantities of oil to produce low, medium
and high energy diets.
Diets were prepared by mixing the dry ingredients in an electric grinder and then
blending in the oils. Water was added and the moist mixture was pelleted using hand
pelletizer. The pellets were oven-dried at 60°C. The dry pellets were crumbled to
appropriate sizes before feeding to the fish.
 K. Samantaray, S.S. Mohan@/Aquaculture 156 (1997) 241-249 243

Table 1
Ingredient and proximate composition of the test diets, expressed as dry wt%
Ingredient Diet number
1 2 3 4 5 6 7 8 9 IO 11 12
Fish meal 24 24 24 27 27 27 32 32 32 37 37 31
Groundnut oil cake 24 24 24 27 27 27 32 32 32 37 37 37
Rice bran 25 25 25 22 22 22 17 17 17 11 11 11
o-Cellulose 23.5 14.5 15.0 20.5 16.5 12.0 15.5 11.5 7.0 11.5 9.5 5.0
Binder” I 1 I 1 1 I I 1 1 1 1 1
Vegetable oil _ 3 7.5 - 3 7.5 - 3 7.5 - 3 7.5
Cod liver oil 0.5 1.5 1.5 0.5 1.5 1.5 0.5 I .5 I .5 0.5 1.5 I .5
Vitaminb and mineral’ 2 2 2 2 2 2 2 2 2 2 2 2
mixture (1 gm each)

Nutrient content

Crude protein 35 35 35 40 40 40 45 45 45 50 50 50
Crude hpid 9 13 17 9 13 17 9 13 17 9 13 17
Digestibel energy 400 440 480 400 440 480 400 440 480 400 440 480
Protein toenergy(P/E) 87.5 79.5 72.9 100.0 90.9 83.3 112.5 102.3 93.8 125.0 113.6 104.2
ratio (mg protein/kcal)

’ Carboxymethyl cellulose.
‘To supply/l00 g diet: Vitamin A (as acetate), 10000 IU; cholecalciferol, 1000 IU; thiamin mononitrate, 10.0
mg, riboflavin, 10 mg; pyridoxine hydrochloride, 60 mg; cyanocobalamin, 30 pg; nicotinamide, 200 mg;
ascorbic acids, 300 mg; cu-tocopheryl acetate, 50 mg: biotin, 0.5 mg.
’ To supply/100 g diet: Calcium phosphate 258 mg; magnesium oxide, 120 mg; ferrous sulphate, 64.08 mg;
manganese sulphate, 4.06 mg; copper sulphate, 6.78 mg; zinc sulphate, 4.40 mg; sodium molybdate, 0.5 mg;
sodium borate, I .76 mg.

2.2. Experimental animal and design

Fingerling Ch. striata of average weight 12 k 2 g (10.0-13.8 g), obtained from a

local supplier, were used for this experiment. The fingerlings underwent a 2-week
conditioning period to adjust to the experimental diet and conditions, prior to the
beginning of the experiment. During this period the fish were fed one of the experimen-
tal diets (diet 1) with the lowest protein level.
The experiment was conducted in two phases in 18 110-l fibre blast tanks each with
its own recirculating system to get three replicates of each treatment, as adequate
numbers of tanks were not available. The flow rate was maintained at 2.0 1 mini ‘. In
phase one diets 1 to 6 were tested and in phase two diets 7 to 12 were tested following
the same protocol for each phase. Each phase of the experiment was continued for 56
days. All tanks were aerated continuously from 08:OO to 15:OO h throughout the
experimental period. Water temperature, dissolved oxygen and pH were recorded daily
and were 28“ _+ 2°C 5.9 to 7.2 and 7.3 to 7.6 mg l-‘, respectively. There was not much
variation in these parameters in the two phases of the experiment. The diurnal light/dark
cycle of 12: 12 h was maintained by supplemental incandescent lighting.
The experimental fish were weighed individually at the beginning and end of the
experiment and batch weighed at weekly intervals for feed adjustments. The fish were
244 K. Samantaray, S.S. Mohanty/Aquaculture 156 (19971 241-249

fed at a rate equalling 3
days a week.
At the beginning of
carcass analysis. At the
feeding trial were killed
wet body wt% per day, twice daily at 09:OO and 16:00 h, seven
the experiment, 12 fingerlings were sacrificed and stored for
end of the experiment, 9 fish (3 from each tank) from each
and frozen immediately for subsequent carcass analysis.

2.3. Analytical methods

Diet ingredients, experimental diets and fish carcasses were analysed following
Association of Official Analytical Chemists (197.5) methods. Moisture was determined
by oven drying at 100°C to constant weight. Crude protein was determined based on
Kjeldahl nitrogen (crude protein = N X 6.25). Samples were extracted with petroleum
ether for 8 h in a Soxhlet extraction apparatus for crude fat determination. Ash content
was determined in a muffle furnace for 6 h at 550°C.
The growth rate (average daily growth percent, %ADG), feed conversion ratio (FCR),
protein efficiency ratio (PER) and apparent net protein utilization percent (%ANPU) or
percent protein deposited were calculated as follows:
Final body weight - initial body weight
%ADG = x 100
days
Dry feed fed in g
FCR =
Wet weight gain in g
Wet weight gain in g
PER =
Dry protein fed in g
Final carcass protein - initial carcass protein
%ANPU or %Protein deposited =
Total dry protein fed
x 100
All data were analyzed using analysis of variance (SPSS/PC program). Multiple
comparisons among means were made with Duncan’s multiple range test (Duncan,
1955).

3. Results

The growth responses of snakehead fingerlings fed the 12 experimental diets with 4
protein levels and 3 energy levels are shown in Table 2. The growth rate was found to
increase with an increase in energy from 440 (9% lipid) to 480 kcal 100 g-’ (17% lipid
level) at the higher protein levels (45-50%) but decrease at the lowest protein level
(35%). At 40% dietary protein, growth rate was found to be highest at 440 kcal 100 g-’
(13% lipid). Of the 12 experimental diets, diets 5 and 9 with energy levels of 440 kcal
(13% lipid) and 480 kcal (17% lipid) at protein levels of 40 and 45, respectively,
showed better growth performance compared to that of other diets (Table 2). However,
K. Samantaray, S.S. Mohanty/Aquadture 156 (1997) 241-249 245
246 K. Samantaray, S.S. Mohanty/Aquaculture 156 (1997) 241-249
 K. Samantaray, S.S. Mohanty/Aquaculture 156 (1997) 241-249 241

diet 5 produced fish more efficiently in terms of protein deposition (%ANPU) when
compared with diet 9 with the higher protein level.
The %ADG was positively correlated to the mean protein intake of fish day -’ (X)
and can be expressed by the equation: %ADG = 5.268 + 0.064X (r = 0.92; P < 0.05).
The trend of changing %ADG in relation to P/E ratio, shown in Table 2, indicates
that the diet at a P/E ratio of 90.9 mg protein kcall’ (diet 5) was optimum for
maximum growth of snakehead fingerlings. The growth remained almost at the same
level with an increase in P/E ratio up to 93.8 mg protein kcal-’ (diet 9). However,
with further increase in P/E ratio to 100 mg protein kcal-’ at the 40% crude protein
level (diet 4) or decrease in P/E ratio to 87 mg protein kcal- ’ at the 35% crude
protein level (diet l), growth rate was reduced significantly.
Diets with lower protein levels (35-40%) produced fish more efficiently in terms of
protein deposition than the diets with higher protein (45-50%) (Table 2). It was also
observed that protein deposition was highest at an energy level of 440 kcal 100 g- ’
(13% lipid) than at the other two energy (400 and 480 kcal 100 g- ’ > or lipid (9 and
17%) levels at 35% dietary protein level. The mean PER and FCR in relation to dietary
protein and energy is shown in Table 2. A significant increase in PER was observed
with an increase in dietary energy from 400 to 480 kcal 100 g-’ at 45% protein and 400
to 440 kcal 100 g-’ at 40% and 50% protein; it decreased with increase in energy at
35% dietary protein. The highest PER and lowest FCR values were obtained at a P/E
ratio of 90.9 mg protein kcal- ’ . The FCR values were consistently poor in this
experiment, more particularly for the fish fed 35% protein diets.
Except for moisture, the carcass composition was found to be affected by changes in
dietary P/E ratio (Table 3). Carcass protein content was found to increase with an
increase in energy from 400 to 440 kcal 100 gg ’ at the lower dietary protein levels
(35-40%).

4. Discussion

The importance of dietary protein level in relation to energy for snakehead fingerlings
is evident in the present study as was found earlier for several other fish species (Garling
and Wilson, 1976; Murai et al., 1985; Lovell, 1989; Das et al., 1991; De Silva et al.,
1991; Garcia-Riera et al., 1993). The snakehead fingerlings grew best with a diet
containing 40% protein, 440 kcal energy and 13% lipid with a P/E ratio of 90.9 mg
protein kcal~ ’ . However, a lower, but not significantly different (P > 0.05) growth was
attained with a diet containing 45% protein, 480 kcal energy, 17% lipid and P/E ratio
of 93.8 mg protein kcall ’ The dietary protein requirement of snakehead fingerlings has
been reported to be 50% (Wee and Tacon, 1982; Wee, 1986; Mohanty and Samantaray,
1997) using isocaloric diets containing 432 kcal digestible energy. But in the present
study, it was observed that, at an optimum P/E ratio, maximum growth can be
obtained at a lower dietary protein level. This indicates that if the dietary energy level is
not maintained by proper use of either lipid or carbohydrate, then protein may be
utilized for energy rather than for growth.
A P/E ratio of 88 mg protein kcal - ’ between protein levels of 24 to 36% has been
reported to be optimum for channel catfish (Garling and Wilson, 1976), 87.6% for Cl.
248 K. Samantaray, S.S. Mohanty/Aquaculture 156 (19971241-249

batruchus (Mukhopadhyay et al., 1986) and 95% at 38% protein level for L. ruhitu (Das
et al., 1991). For snakehead fingerlings, in the present study, the optimum P/E ratio
was found to be 90.9 mg protein kcal-’ at 40% protein level based on growth
performance, FCR, PER and %ANPU.
The minimum level of lipid required for maximum growth, survival and feed
conversion of snakehead fingerlings has been reported to be 6% (Boonyaratpalin, 198 1).
In common, carp growth was not depressed when 18% dietary lipid was fed (Jauncey,
1982). Channel catfish, however, showed growth depression when the dietary lipid level
was increased from 12% to 16% of the dry diet (Dupree et al., 1979). Red tilapia can
effectively utilize a lipid level of 18% (De Silva et al., 1991). No growth-depressing or
pathological effects were observed when high levels of dietary lipid were fed to rainbow
trout (Garcia-Riera et al., 1993). From the present study, it can be concluded that a lipid
level of 13% can be effectively utilized by snakehead fingerlings.
The FCR and PER are known to decrease and increase, with increasing dietary
protein content up to a level that gives optimum growth, respectively (Wee and Tacon,
1982; Wee, 1986). Results of the present study indicate that dietary energy at all protein
levels influence PER. Snakehead are slow feeders and thus nutrient leaching occurred in
the present study and may be the cause for the poor FCR.
The changes in carcass composition in relation to dietary protein and energy levels
were less consistent. The protein level appeared to increase at the lower dietary protein
(35-40%) and energy (400-440 kcal) levels.
This study has demonstrated the capability of snakehead fingerlings to spare protein
and the degree to which the lipid content in the diet can be effectively used to obtain
optimal growth. Diets containing about 90.9 mg protein kcall’ energy were adequate
for snakehead fingerlings for maximum growth within certain dietary protein and energy
concentrations.

Acknowledgements

The financial assistance of International Foundation of Science (Grant No. A/2206-1)

for this study is gratefully acknowledged.

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