See

discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/264609004

Food consumption and digestive enzyme

activity of Clarias batrachus exposed to various
temperatures

Article in Aquaculture Nutrition · June 2014

DOI: 10.1111/anu.12072

CITATIONS READS

2 35

5 authors, including:

Tauqueer Ahmad Bronson Kumar Khangembam

University of Delhi University of Delhi
3 PUBLICATIONS 2 CITATIONS 5 PUBLICATIONS 9 CITATIONS

SEE PROFILE SEE PROFILE

Jaigopal Sharma Rina Chakrabarti

Barkatullah University University of Delhi
45 PUBLICATIONS 236 CITATIONS 100 PUBLICATIONS 1,010 CITATIONS

SEE PROFILE SEE PROFILE

All in-text references underlined in blue are linked to publications on ResearchGate, Available from: Rina Chakrabarti
letting you access and read them immediately. Retrieved on: 10 October 2016
 Aquaculture Nutrition
2014
..........................................................................................
1 1
1 2
Aqua Research Lab, Department of Zoology, University of Delhi, Delhi, India;
nological University, Delhi, India
The effect of temperature on the food consumption rate
and the digestive enzyme activities of Clarias batrachus
(80.60
5.34 g) were evaluated. Fish were exposed to six
different temperatures of 10, 15, 20, 25, 30 and 35 °C fol-
lowing an acclimation temperature of 25 °C. The rate of
temperature change was 2 °C day1. Highest food con-
sumption was recorded at 25 °C. It gradually reduced with
decreasing water temperature. Food consumption rate was
significantly (P < 0.05) lower at 10 °C compared with other
treatments. Hence, 46.67, 8.20–23.58 and 1.02–6.15%
reduced food consumptions were recorded in groups
exposed at 10, 15 and 20 °C temperatures, respectively,
compared with the 25 °C. The consumption rate was not
affected in fish exposed at 30 and 35 °C. Total protease,
trypsin and chymotrypsin activities were significantly
(P < 0.05) higher in fish exposed at 25 °C compared with
others. Lipase activity was significantly (P < 0.05) higher in
fish exposed at 30 °C compared with others. Lowest enzyme
activities were recorded at 10 °C. Water temperature below
25 °C affected the food consumption and digestive enzyme
activities in fish that served as indicators of stress in fish.
KEY WORDS: Clarias batrachus, digestive enzymes, food con-
sumption, temperature
Received 6 December 2012; accepted 25 February 2013
Correspondence: R. Chakrabarti, Aqua Research Lab, Department of
Zoology, University of Delhi, Delhi 110007, India. E-mail: aquaresearch
lab@yahoo.co.in
Among the various environmental factors, temperature
plays critical role in the life of poikilothermic aquatic
organisms. Physiological processes of fish viz. food
..............................................................................................
ª 2014 John Wiley & Sons Ltd
doi: 10.1111/anu.12072
1 2 1
Department of Biotechnology, Delhi Tech-
consumption, digestion, immune system, etc. are influenced
by the water temperature where they live. The quantifica-
tion of food consumption by fish is a parameter central to
fish ecology, both for the assessment of wild fish stocks,
where food availability may limit growth and reproduction,
as well as in aquaculture, where it is often desirable to
maximize feed intake without wasting expensive feed to
maximize profit (Richter et al. 2004). Consumption esti-
mates often constitute important parameters in aquatic eco-
system models, such as the ECOPATH model (Christensen
& Pauly 1992), where they help to govern energy flows
between compartments and trophic levels.
Among the factors governing growth of fish, food is the
driving force, temperature is a rate-controlling factor and
fish size a scaling factor (Brett & Groves 1979). When the
effects of such factors on feeding, metabolism and growth
are known, models can be used for estimations of feeding
in natural populations instead of direct measurements of
food intake in the field (Fonds et al. 1992). Rate of feed-
ing, metabolism and growth are affected not only by the
availability of food but also directly by water temperature.
Even ingestive variations were reportedly influenced by
water temperature (Lagler et al. 1977). The temperature
coefficient (Q10) represents the degree of sensitivity of an
organism to temperature (Diaz-Iglesia 1988).
Digestion of ingested food is essential to meet the energy
requirement of the fish. The ability of any fish to digest a
given diet and absorption of its nutrients depends on the
presence and the quality of digestive enzymes (Chakrabarti
& Rathore 2009). As protein utilization is fundamental to
growth, proteases have an important role to play in the
larval fish as in the adult (Kumar et al. 2007). The
digestive proteases of different species showed variations
(Chakrabarti & Sharma 2005), which may influence their
digestive capability and feeding habits. Among the various
nutritional components of prepared diet, protein source is
the most expensive that can be replaced by lipid source.
Therefore, lipase plays significant role in the digestion of
lipid-rich prepared food.
The Global climate is changing rapidly, and significant
climate shifts will affect freshwater fish communities and
fisheries. Global warming is now 0.6 °C in the past three
decades and 0.8 °C in the past century (Hansen et al.
2006). Climate change will present fishes with new environ-
ments through a number of mechanisms not necessarily
operating independently with the possibility of synergy, or
an interaction between them is highly probable. All fresh-
water fishes are exotherms that cannot regulate their body
temperature through physiological means (Moyle & Cech
2004) The increasing global temperature can affect individ-
ual fish by altering physiological functions such as thermal
tolerance, growth, metabolism, food consumption, repro-
ductive success and the ability to maintain internal homo-
eostasis in the face of a variable external environment (Fry
1971). Fish populations that are faced with changing ther-
mal regimes may increase or decrease in abundance, experi-
ence range expansions or contractions, or face extinction
(Ficke et al. 2007).
The Asian catfish, Clarias batrachus (Linnaeus, 1758),
popularly known as magur is widely distributed throughout
Indian subcontinent. It is characterized by its flesh quality,
less bones, rapid growth and high marketability that make
it a good candidate for aquaculture (Singh et al. 2009). Lit-
tle information is available on the effect of temperature on
the performances of this economically important species.
The present investigation attempts to establish relationship
between water temperature and food consumption rate of
Clarias batrachus in the present scenario of climate change.
The effect of temperature on the digestive enzyme activities
was also recorded. Digestion of ingested food is essential to
meet the energy requirement of the fish. Food consumption
rate and digestive enzyme profile are good indicators of
health status of fish. These are important parameters of
aquaculture, where it is often desirable to maximize feed
intake without wasting expensive feed to maximize profit.
Asian catfish Clarias batrachus (popularly known as
magur) were obtained from Chatterjee Brothers’ Fish
Farm, West Bengal. Fish (initial weight: 80.60
5.34 g)
were randomly distributed in glass aquaria (five fish 50 L
aquarium) inside the wet laboratory facility and were accli-
mated at 25 °C for 21 days. This duration of acclimation
helped to avoid handling stress. Fish were maintained at
12 h: 12 h dark and light regime throughout the study per-
iod. Each aquarium was connected to a filtration unit and
a cooling/heating unit. Aquaria were covered with trans-
parent fibre glass sheets with two holes for easy passage of
inlet and outlet pipes to the cooling/heating units. Water
from fish culture unit was constantly filtered through the
filtration unit. This water consequently entered the cooling
or heating unit and finally back to the fish culture unit.
Heating and cooling units remained non-functional during
acclimatization period. pH and dissolved oxygen level ran-
ged from 7.8–8.2 and 5.8–6.1 mg L1, respectively, during
the study period. As the ambient temperature was
25
2 °C, fish were acclimated at 25 °C to give natural
environmental condition to the fish.
Fish were exposed simultaneously to six different tempera-
tures: 10, 15, 20, 25, 30 and 35 °C. Three replicates were
used for each temperature. 25 °C temperature was consid-
ered as ambient temperature (control). Each experimental
temperature was achieved at a rate of change of 1 °C per
12 h starting from acclimation temperature 25 °C. This
required 2.5–7.5 days to reach the assigned temperatures.
The range of temperature selected in this experiment was
based on seasonal variation of environmental temperature
in India. The total experimental exposure period was
7 days after reaching the assigned temperature in all treat-
ments, except 10 °C. All fish died at 10 °C within 4 days.
There was no mortality of fish in other temperature
groups.
Fish were fed with practical diet, prepared in our labora-
tory at the rate of 50 g kg1 body weight of fish (5%)
(Table 1) throughout the study period. The proximate com-
position of the prepared diet was analysed through SGS
India Pvt. Ltd. Crude protein, fat and moisture contents
were analysed following the standard methods (BIS 1975).
Total carbohydrate and free fatty acids were analysed by
following the methods of AOAC (1995) and AOCS (2009),
respectively. Amino acid profile of diet was evaluated using
6-aminoquinolyl-N-hydroxylsuccinimidyl carbamate (AQC
reagent).
The total amount of food was divided in two parts and
delivered at 9.00 a.m. and 5.00 p.m. A pilot scale study
..............................................................................................
Aquaculture Nutrition ª 2014 John Wiley & Sons Ltd
 Table 1 Ingredients, proximate composition and amino acid profile
of experimental diet
Ingredients (g kg1)
Dry fish powder
Wheat flour
Cod liver oil
Vitamin-mineral premix
Proximate composition (g kg1)
Moisture
Crude protein
Crude fat
Total carbohydrate
Free fatty acid
Amino acid profile (g kg1)
Histidine
Isoleucine
Leucine
Lysine
Methionine
Phenylalanine
Threonine
Valine
Arginine
was performed during acclimation period to establish the
optimal feeding rate. Fish were fed with 8%, 7%, 6%, 5%
and 4% of body weight. It was found that 5% feeding
scheme was suitable for this size group of magur and for
maintenance of water quality. The food consumption rate
of fish was monitored regularly after each feeding. For this
purpose, unconsumed food was siphoned from individual
aquarium after three hours of feeding. The collected food
was dried, and the weight of the dry food was recorded.
The amount of food consumed during each meal was cal-
culated as the difference between the mass of the food
offered and that of the uneaten food after drying. Food
consumption that was calculated for individual fish after
each meal is expressed as g fish per meal.
Fish were collected from each aquarium (two fish per aquar-
ium) after 12 h of reaching assigned temperature to study
the effect of acute stress on the digestive enzyme profile of
magur. The fish were dissected on a glass plate maintained
at 0 °C. The digestive tract of individual fish was collected,
rinsed with chilled distilled water, blot-dried and weighed.
Then, the sample was homogenized with chilled distilled
water (1:10) to maintain neutral pH of extract as this extract
was used for various enzyme assays at different pHs. The
homogenate was centrifuged at 10000 9 g for 15 min at
4 °C. The supernatant was used for analysis. Total soluble
protein was measured using the method of Bradford (1976).
..............................................................................................
Aquaculture Nutrition ª 2014 John Wiley & Sons Ltd
Bovine serum albumin (Sigma, St Louis, MO, USA) was
used as a standard against the sample protein (1 mg mL1).
Total protease activity was measured using 1% azocasein
583.3
(Sigma, St Louis, MO, USA) as substrate in 50 mM Tris-
402.7
10.0 HCl, pH 7.5 (SRL, Mumbai, India) following the method
4.0 of Garcia-Carreno (1992). The change of absorbance was
recorded at 366 nm using UV-1601 Spectrophotometer
23.6
438.7
(Shimadzu, Japan). The enzyme activity was recorded in
93.2 Units mg protein1 min1.
364.4 Trypsin and chymotrypsin activities were measured using
656.8
Na-Benzoyl-DL-arginine p-nitroanilide hydrochloride (BAP-
12.0 NA) and N-Succinyl-Ala-Ala-Pro-Phe p-nitroanilide (SAPNA)
19.0 (Sigma, St Louis, MO, USA), as substrate, respectively.
32.0
Change of absorbance was recorded under kinetic mode for
32.0
12.0 3 min at 410 nm (Erlanger et al. 1961). Activity units were
18.0 expressed as change in the absorbance mg protein1 min1.
16.0 Activity units were calculated by the following equation:
23.0
23.0
Activity units


Abs410 min1  1000  mL of reaction mixture (1)
¼
8800  mg protein in reaction mixture
The molar extinction coefficient of p-nitroanilide is 8800.
Lipase activity was measured according to the methods
of Winkler & Stuckmann (1979). The principle of the assay
is the colorimetric estimation of p-nitrophenol released
because of enzymatic hydrolysis of p-nitrophenyl palmitate
(Sigma, St Louis, USA) at 410 nm. One enzyme unit was
defined as 1 lmol of p-nitrophenol enzymatically released
from the substrate mL1 min1. The extinction coefficient
of p-nitrophenol is 15000 cm2 mg1.
Temperature coefficient (Q10) Q10 values for different
enzyme activities were calculated between different temper-
atures using the Van’t Hoff equation (Hochachka &
Somero 2002):
Q10 ¼ ðR2 =R1 Þ10=ðT2 T1 Þ (2)
Where T1 and T2 represent two different temperatures
(T2 > T1); R1 and R2 represent enzyme activity at T1 and
T2, respectively.
Data were compiled as means
SE. All data were analy-
sed using one-way analysis of variance (ANOVA) and regres-
sion analysis using SPSS software 19.0. Statistical
significance was accepted at P < 0.05 level.
 swimming activity might influence the food consumption
rate of fish.
In 15 °C temperature treatment, the food consumption
rate was reduced by 24% on day 4 of reaching this temper-
There was no significant (P > 0.05) difference in average
ature. Then, the consumption rate gradually increased.
weight of fish (95
4 g) after 21 days of acclimation at
There was 16% recovery of food consumption rate on day
25 °C. The average weight of magur exposed at 15 and
7 compared with the day 4 of the same treatment. The
20 °C was 90
7 and 91
5 g, respectively, after 7 days
food consumption rate was significantly (P < 0.05) lower at
of exposure at the assigned temperature. The average
15 °C compared with 20–35 °C groups (Fig. 2). A four-
weight of fish at 25, 30 and 35 °C was 97
2.7, 98
4
degree polynomial relationship (Fig. 2) was found between
and 95
7 g, respectively, after 7 days of exposure at the
the temperature and food consumption rate of magur
assigned temperature. In 10 °C treatment, as all fish died
(R² = 0.92) in this treatment.
after four days of reaching assigned temperature, there
In the 20 °C temperature exposed group, 5% reduced
were no data for 7 days exposure.
food consumption was recorded on first day of reaching
this temperature compared with the consumption rate at
25 °C. Food consumption rate of fish showed six-degree
polynomial relationship (Fig. 3) with the decreasing water
There was no significant (P > 0.05) difference in food con-
temperature (R² = 0.94) in this group.
sumption rate (1.95
0.01 g fish1) of magur during accli-
The highest food consumption of magur was recorded at
matization (25 °C). As the water temperature gradually
25 °C and the rate gradually decreased; lowest consump-
decreased in treatments 10, 15 and 20 °C, the rate of food
tion was found at 10 °C. A polynomial three-degree rela-
consumption of individual fish also decreased. Significantly
tionship was found between the culture temperature and
(P < 0.05) lower food consumption rate was recorded at
food consumption rate of magur regardless of the treat-
10 °C compared with the fish exposed to other tempera-
ment (Fig. 4). There was no change in food consumption
tures (Fig. 1). Fish consumed 46.67% less food on first and
rate of fish exposed to 30 and 35 °C compared with the fish
second days of reaching 10 °C compared with the acclima-
maintained at 25 °C.
tion one. Fish survived two more days in this treatment
without feeding. A four-degree polynomial relationship was
found between the water temperature and the food con-
sumption rate of fish (R² = 0.99). Fish showed lethargic
Total protease activity was significantly (P < 0.05) higher
swimming behaviour at 10–15 °C temperatures. Fish were
in fish assigned at 25 °C compared with the fish of other
mostly remaining at the bottom of the aquarium. Less
groups. Lowest protease activity was found in 10 °C

Figure 1 Effect of temperature on the food consumption rate of
Clarias batrachus (n = 3). Water temperature gradually decreased
from 25 to 10 °C. Bar indicates the food consumption of fish in
different days at a particular temperature.
Figure 2 Effect of temperature on the food consumption rate of
Clarias batrachus (n = 3). Water temperature gradually decreased
from 25 to 15 °C. Bar indicates the food consumption of fish in
different days at a particular temperature.
..............................................................................................

Aquaculture Nutrition ª 2014 John Wiley & Sons Ltd

 Figure 5 Total protease activity found in the digestive tissue of
Figure 3 Effect of temperature on the food consumption rate of
Clarias batrachus exposed to various temperatures (n = 3).
Clarias batrachus (n = 3). Water temperature gradually decreased
from 25 to 20 °C. Bar indicates the food consumption of fish in
different days at a particular temperature.

Figure 6 Trypsin activity found in the digestive tissue of Clarias

batrachus exposed to various temperatures (n = 3).

Figure 4 Food consumption of Clarias batrachus showed polyno-

mial (three degree) relationship with decreasing temperature from
the acclimation temperature (25 °C).

assigned group (Fig. 5). A 3.90–5.38-fold decreased prote-

ase activity was recorded in fish exposed at 20–10 °C com-
pared with the enzyme activity found at 25 °C. Whereas
1.31–1.74-fold reduced activity was found in groups
exposed to 30–35 °C compared with the 25 °C treatment.
A similar trend was also found with trypsin activity. The
enzyme activity was significantly (P < 0.05) higher in the
group maintained at 25 °C compared with others (Fig. 6).
This group was followed by groups exposed at 35 and Figure 7 Chymotrypsin activity found in the digestive tissue of
30 °C. Trypsin activity was lowest in the fish exposed at Clarias batrachus exposed to various temperatures (n = 3).
10 °C. Like trypsin, chymotrypsin activity was significantly
(P < 0.05) higher in the fish maintained at 25 °C compared with others. This was followed by the group exposed to
with the other groups (Fig. 7). Lipase activity was signifi- 20 °C. Lowest lipase activity was recorded in fish exposed
cantly (P < 0.05) higher in fish exposed at 30 °C compared at 10 °C (Fig. 8).
..............................................................................................

Aquaculture Nutrition ª 2014 John Wiley & Sons Ltd

 et al. 1989; Bendiksen et al. 2002), growth (Brown et al.
1989) and activity (Clark et al. 1995). Kumari et al. (2006)
reported the decreased growth rate and increased suscepti-
bility to opportunistic pathogens in magur during winter.
Reduced food consumption results into poor growth in fish
during winter months. The average weight of magur was
found to be reduced at low temperature (10–20 °C),
whereas fish at ambient and higher temperature gained
weight in the present study. Foraging particularly mediates
individual performance, as food acquisition is required to
fuel all biological functions (Godin 1997). An individual
that ingests higher quantities of food, for example, is likely
Figure 8 Lipase activity found in the digestive tissue of Clarias to have higher rates of growth and better body condition,
batrachus exposed to various temperatures (n = 3). thereby supporting their survival and fecundity (Lambert
et al. 2000). Atlantic salmon parr responded to the temper-
The temperature coefficient (Q10) values for different ature fluctuation with reduced feed intake (Folkedal et al.
enzymes at various temperature ranges (10 °C interval) 2010).
were calculated (Table 2). A temperature-dependent There was no change in food consumption rate of fish
enzyme activities were found for total protease, chymotryp- exposed at 30 and 35 °C compared with the fish main-
sin and lipase at the temperature range 10–20, 15–25 and tained at 25 °C. The normal activity of the fish at 30 and
20–30 °C as the Q10 values were >1.0. Temperature-depen- 35 °C influenced the food consumption of fish. Although
dent trypsin activities were found at temperature range increased temperature can increase metabolic rate and
10–20, 15–25 °C. Highest Q10 values for total protease, activity levels (Brett 1979, 1995; Jobling 1994; Taylor et al.
trypsin and chymotrypsin were obtained at 15–25 °C, 1997; Biro et al. 2007, 2010), a common observation is that
whereas for lipase it was at 10–20 °C. feeding rate declines once the optimal temperature exceeds.
Our results clearly showed that the food consumption
and digestive enzyme activities were affected by decreased
temperature. High stressful conditions were observed at
Temperature of aquatic environment is important for temperature 10 °C. At this temperature, the fish showed
ensuring survival of aquatic animals, and failure to adapt less appetite for feed. In common carp, Cyprinus carpio
temperature fluctuations is generally ascribed to the inabil- feed conversion efficiency decreased with low temperatures
ity of fish to respond physiologically. In the present study, (Goolish & Adelman 1984), which might be result of
reduced food consumption was recorded with decreasing enzyme kinetics (Singh et al. 2009). The study of digestive
water temperature, which was corelated with the reduced enzyme activity of magur in the present study showed that
swimming activity of the fish. Reduced feeding appears to enzymes associated with protein and lipid digestion were
be a behavioural tactic to maximize energetic efficiency at affected by low temperature of 10–20 °C. At 30 and 35 °C,
high temperature (Brett 1971). Fish are exposed to lower enzyme activities associated with protein digestion (total
temperatures during the winter, and these temperatures protease, trypsin and chymotrypsin) were reduced com-
have a significant negative impact on fish appetite (Brown pared with the fish maintained at acclimation temperature
25 °C, but the lipase activity was highest at 30 °C. Dehad-
Table 2 Values of Q10 for different enzyme activities between dif- rai et al. (1985) reported that magur grew well up to 32 °C
ferent temperatures temperature. Fish were seen to be under stress at 35 °C,
and mortality of fish started at 38 °C. The effects of the
Temperature interval (°C)
temperature on enzyme activity were quantified by calculat-
Enzyme 10–20 15–25 20–30 25–35 ing the temperature coefficient or Q10 of the process. The
Total protease 1.22 3.82 2.51 0.75 physiological processes such as rates of respiration and
Trypsin 1.52 2.89 0.87 0.49 enzymatic process, Q10 values near 2.0 or slightly higher,
Chymotrypsin 1.42 3.09 1.27 0.65 are observed when thermal effects are studied within the
Lipase 2.12 1.31 1.12 0.65
species’ physiological range of body temperatures
..............................................................................................

Aquaculture Nutrition ª 2014 John Wiley & Sons Ltd

 (Hochachka & Somero 2002). All enzymes except lipase
had a higher Q10 value for the temperature range of 15–
25 °C than 10–20 °C. At the temperature (up to 35 °C)
above the acclimation one (25 °C), Q10 values were <1.0,
which indicated the temperature-independent reaction.
Singh et al. (2009) suggested that C. batrachus fry fed with
diet having 360 g kg1 protein level and reared in the
water temperatures 28 and 32 °C can attain maximum
body weight. In the present study, Clarias batrachus
showed unaffected food consumption (400 g kg1 protein
content) rate between the temperature range of 25–35 °C.
Being a tropical species, magur showed adaptability to the
higher temperatures compared with the lower temperature
that was confirmed in the Q10 value.
Reduced food consumption and digestive enzyme activity
are sensitive indicators of stress in fish. This physiological
stress leads to the immunosuppression in fish. Proper care
should be taken during the winter months in the pond cul-
ture as food consumption of fish is reduced. To improve the
physiological condition specific food ingredient viz. immu-
nostimulant, antioxidant, etc. may be included in the com-
mercial diet of fish after observing the decreased food intake
in the species. Food contributes immensely in the production
cost of aquacultural species. Simultaneously, excess food
leads to water quality deterioration, which results into out-
break of disease and mortality of fish. Therefore, a tempera-
ture-dependent feeding rate for each developmental stage of
individual species is required for cost-effective production.
The present study has provided the base line data for the
development of husbandry conditions of Clarias batrachus.
This study has been carried out with the financial support
from Indian Council of Agricultural Research, ICAR, New
Delhi (NFBSFARA project, AS-2001/2010-11). The proxi-
mate composition of the prepared diet was analysed
through SGS India Pvt. Ltd., SGS House, 4B Adi Shank-
aracharya Marg, Vikhroli (West), Mumbai 400083, India.
AOAC Association of Official Analytical Chemists (1995) Official
Methods of Analysis of AOAC. 16th edn, Vol 1, Arlington, Vir-
ginia, USA.
AOCS American Oil Chemists’ Society (2009) Preparation of
methyl esters of fatty acids. In: Official Methods and Recom-
..............................................................................................
Aquaculture Nutrition ª 2014 John Wiley & Sons Ltd
mended Practices of the American Oil Chemists’ Society (Fire-
stone, D. ed.), 6th edn, pp. Ab 5–49. AOCS Press, Champaign.
Bendiksen, E., Jobling, M. & Arnesen, A.M. (2002) Feed intake of
Atlantic salmon parr Salmo salar L. in relation to temperature
and feed composition. Aquacult. Res., 33, 525–532.
Biro, P.A., Post, J.R. & Booth, D.J. (2007) Mechanisms for cli-
mate-induced mortality of fish populations in whole-lake experi-
ments. Proc. Natl Acad. Sci. USA., 104, 9715–9719.
Biro, P.A., Beckmann, C. & Stamps, J.A. (2010) Small within-day
increases in temperature affects boldness and alters personality
in coral reef fish. Proc. Bio. Sci., 277, 71–77.
BIS Bureau of Indian Standards (1975) Indian Standard Methods
of Test for Animal Feeds and Feeding Stuffs: Part I General
Methods. Standard No. IS: 7874, BIS Publications, India.
Bradford, M.M. (1976) A rapid and sensitive method for the quan-
titation of microgram quantities of protein utilizing the principle
of protein-dye binding. Anal. Biochem., 72, 248–254.
Brett, J.R. (1971) Energetic response of salmon to temperature. A
study of some thermal relations in the physiology and freshwater
ecology of sockeye salmon (Oncorhynchus nerka). Am. Zool., 11,
99–113.
Brett, J.R. (1979) Environmental factors and growth. In: Fish
Physiology (Hoar, W.S., Randall, D.J. & Brett, J.R. eds), Vol. 8,
pp. 599–675. Academic Press, London.
Brett, J.R. (1995) Energetics. In: Physiological Ecology of Pacific
Salmon (Groot, C., Margois, L. & Clarke, W.C. eds), pp. 3–68.
University British Columbia Press, Vancouver.
Brett, J.R. & Groves, T.D.D. (1979) Physiological energetics. In:
Physiological Energetic (Hoar, W.S., Randal, D.J. & Brett, J.R.
eds), pp. 279–352. Academic Press, New York.
Brown, J.A., Pepin, P., Methven, D.A. & Somerton, D.C. (1989)
The feeding, growth and behaviour of juvenile cod, Gadus mor-
hua L., in cold environments. J. Fish Biol., 35, 373–380.
Chakrabarti, R. & Rathore, R.M. (2009) Ontogenic changes in the
digestive enzyme pattern and characterization of proteases in
Indian major carp Cirrhinus mrigala. Aquacult. Nutri., 16, 569–
581.
Chakrabarti, R. & Sharma, J.G. (2005) Digestive physiology of
fish larvae during ontogenic development: a brief overview.
Indian J. Anim. Sci., 75, 1337–1347.
Christensen, V. & Pauly, D. (1992) ECOPATH II-a software for
balancing steady-state ecosystem models and calculating network
characteristics. Ecol. Model., 61, 169–185.
Clark, D.S., Brown, J.A., Goddard, S.J. & Moir, J. (1995) Activity
and feeding behaviour of Atlantic cod (Gadus morhua) in sea
pens. Aquaculture, 131, 49–57.
Dehadrai, P.V., Yusuf, K.M. & Das, R.K. (1985) Package and
practices for increasing production of air breathing fishes. In:
Aquaculture Extension Manual, New Series, Information and
Extension Division of CIFRI, pp. 1–14. Indian Council of Agri-
culture Research, Barrackpore.
Diaz-Iglesia, E. (1988) Aspectos de la Fisiologia de Animals Acu-
aticos. Editorial Pueblo y Educacion, La Habana, Cuba. pp.119.
Erlanger, B.F., Kokowski, N. & Cohen, W. (1961) The prepara-
tion and properties of two chromogenic substances of trypsin.
Arch. Biochem. Biophysiol., 95, 271–278.
Ficke, A.D., Myrick, C.A. & Hansen, L.J. (2007) Potential impacts
of global climate change on freshwater Fisheries. Rev. Fish Biol.
Fisheries., 17, 581–613.
Folkedal, O., Torgersen, T., Nilsson, J. & Oppedal, F. (2010)
Habituation rate and capacity of Atlantic salmon (Salmo salar)
parr to sudden transitions from darkness to light. Aquaculture,
307, 170–172.
Fonds, M., Cronie, R., Vethaak, A.D. & van der Puyl, P. (1992)
Metabolism, food consumption and growth of plaice (Pleuronec-
tes platessa) and flounder (Platichthys flesus) in relation to fish
size and temperature. Neth. J. Sea Res., 29, 127–143.
Fry, F.E.J. (1971) The effect of environmental factors on the phys-
iology of fish. In: Fish Physiology: Environmental Relations and
Behavior (Hoar, W.S. & Randall, D.J. eds), pp. 1–98, Academic
Press, New York.
Garcia-Carreno, F.L. (1992) The digestive proteases of langostilla
(Pleuroncodes planipes, decapoda): their partial characterization,
and the effect of feed on their composition. Comp. Biochem.
Physiol. B., 103, 575–578.
Godin, J.G.J. (1997) Behavioural Ecology of Teleost Fishes.
Oxford University Press, USA.
Goolish, E.M. & Adelman, I.R. (1984) Effects of ration size and
temperature on the growth of juvenile common carp (Cyprinus
carpio L.). Aquaculture, 36, 27–35.
Hansen, J., Sato, M., Ruedy, R., Lo, K., Lea, D.W. & Medina-Eli-
zade, M. (2006) Global temperature change. Proc. Nat. Acad.
Sci. USA., 103, 14288–14293.
Hochachka, P. & Somero, G. (2002) Biochemical Adaptation,
Mechanism and Physiological Evolution. pp. 480, Oxford Uni-
versity Press, New York.
Jobling, M. (1994) Fish Bioenergetics. Chapman & Hall, London.
Kumar, S., Fernando, L.G., Chakrabarti, R., del Toro, M.A.N. &
Cordova-Murueta, J.H. (2007) Characterization of digestive pro-
teases of three carps Catla catla, Labeo rohita and Hypophthal-
michthys molitrix and in vitro digestibility study. Aquacult. Nutr.,
13, 1–9.
Kumari, J., Sahoo, P.K., Swain, T., Sahoo, S.K., Sahu, A.K. &
Mohanty, B.R. (2006) Seasonal variation in the innate immune
parameters of the Asian catfish Clarias batrachus. Aquaculture,
252, 121–127.
Lagler, K.F., Bardach, J.E., Miller, R.R. & Passino, D.R.M.
(1977) Ichthyology. 2nd edn, Wiley, New York.
Lambert, Y., Dutil, J.D. & Ouellet, P. (2000) Nutritional condition
and reproductive success in wild fish populations. In: Proceedings
of the 6th International Symposium of Reproductive Physiology
of Fish (Norberg, B., Kjesbu, O.S., Taranger, G.L., Andersson,
E. & Stefansson, S.O. eds.), pp. 77–84. John Grieg, Bergen.
Moyle, P.B. & Cech, J.J. (2004) Fishes: An Introduction to Ichthy-
ology. 5th edn. Prentice Hall, Englewood Cliffs.
Richter, H., L€uckst€adt, C., Focken, U. & Becker, K. (2004) Some
mathematical considerations in estimating daily ration in fish
using food consumption models. Ecol. Model., 171, 381–393.
Singh, R.K., Desai, A.S., Chavan, S.L. & Khandagale, P.A. (2009)
Effect of water temperature on dietary protein requirement,
growth and body composition of Asian catfish, Clarias batrachus
fry. J. Therm. Biol., 34, 8–13.
Taylor, E.W., Egginton, S., Taylor, S.E. & Butler, P.J. (1997) Fac-
tors which may limit swimming performance at different temper-
atures. In: Global Warming: Implications for Freshwater and
Marine Fish (Wood, C.M. & McDonald, D.G. eds.), pp. 105–
133. Cambridge University Press, Cambridge.
Winkler, U.K. & Stuckmann, M. (1979) Glycogen, hyaluronate,
and some other polysaccharides greatly enhance the formation
of exolipase by Serratia marcescens. J. Bacteriol., 138, 663–670.
..............................................................................................
Aquaculture Nutrition ª 2014 John Wiley & Sons Ltd