Effect of Ration Level and Feeding Frequency on Growth, Nutrient Utilization and Body Composition of Juvenile Black Fin Sea Bream, Acanthopagrus berda (Forsskal 1775)

Abdur Rahim1, Ghulam Abbas1*, Lorenzo Gallus2, Sara Ferrando2, Muhammad Hafeez-ur-Rehman3, Abdul Ghaffar4 and Abdul Mateen5

1Centre of Excellence in Marine Biology, University of Karachi, Karachi-75270, Pakistan

2Department of Earth, Environment and Life Sciences (DISTAV), University of Genoa, Italy

3Department of Fisheries and Aquaculture, University of Veterinary & Animal Sciences, Lahore, Pakistan

4Department of Life Sciences, The Islamia University of Bahawalpur, Bahawalpur, Pakistan

5Department of Zoology, Wildlife and Fisheries, University of Agriculture, Faisalabad, Pakistan

ABSTRACT

In this study, effect of different feeding level and frequency on growth performance, nutrient utilization and body composition of juvenile black fin sea bream, Acanthopagrus berda (weight 23.5g−125.8 g) were investigated. Fish were randomly distributed in rectangular glass tanks (150 liter) and were fed at six ration levels of 2.0, 2.5, 3.0,3.5, 4.0 and 4.5% body weight per day (BW d−1) and four feeding frequency i.e., once, twice, thrice and four times per day. Fish in all replicates were fed with diet comprising 40% protein and 20% lipid for 75 days. Higher percent weight gain (% WG), best feed conversion ratio (FCR) and specific growth rate (SGR) were recorded at ration level from 2.5 to 4.5% BW d−1 and feeding frequency of three to four times daily. The moisture, protein and ash contents of whole body of the fish were not significantly (P>0.05) affected by feeding frequency. The highest lipid contents were observed in fish at feeding frequency of three to four times daily. The condition factor (CF), viscerosomatic index (VSI) and hepatosomatic index (HSI) were significantly (P< 0.05) higher in fish at ration levels from 2.5 to 4.5% BW d−1 and feeding frequency of three to four times daily. These results concluded that the optimum ration level and feeding frequency of the juvenile sea bream A. berda (weight 23.5g to 125.8 g) are 2.5% BW d−1 and three times daily, respectively, under similar culture conditions.

Article Information

Received 29 January 2016

Revised 24 May 2016

Accepted 02 August 2016

Available online 10 February 2017

Authors’ Contributions

GA conceived and designed the study and wrote the article. AR executed all the experimental work. MHR analyzed feed components. AM analyzed fish meat samples. AG statistically analyzed the data. SR and LG helped in preparation of manuscript.

Key words

Sea bream (Acanthopagrus berda), Feeding frequency, Ration level, Nutrient utilization, Growth.

* Corresponding author: abbas.cemb@yahoo.com

0030-9923/2017/0002-517 $ 8.00/0

Copyright 2016 Zoological Society of Pakistan

DOI: http://dx.doi.org/10.17582/journal.pjz/2017.49.2.557.563

INTRODUCTION

 

Sea breams are the most important commercial fishes constituting a significant demersal fish resource of Asia, Africa and Europe (Mourente and Bell, 2006; Abbas et al., 2015; Rahim et al., 2015). Black fin sea bream, Acanthopagrus berda has high market value in Pakistan due to its best taste and public demand (Abbas et al., 2011; Anonymous, 2012). But, captive stocks of sea bream have been drastically decreased than other groups of fishes like snappers and grunts in the last decade in our waters due to over exploitation and mismanagement of fisheries resources (Anonymous, 2012). Feasible remedy for government and farmers in this regard is the culture of this species on commercial scale in order to fulfill the protein needs of the country (Abbas et al., 2015; Rahim et al., 2015). Fish feed is considered as an important component for such type of aquaculture development on commercial scale (Abbas et al., 2011). Feed contributes considerably to the running cost of any culture system. In addition, determination of daily ration is important for sustainable growth and suitable feed conversion of fish (Wang et al., 2007; Aderolu et al., 2010;

Abbas et al., 2015). Ration size and frequency varies from species to species and also depend on many physical factors like temperature (Russel et al., 1996). Moreover, feeding frequency and ration level greater than optimum level may decrease digestive efficiency and also have adverse effect on the water quality (Andrews and Page, 1975; Ng et al., 2000). Therefore, an optimum ration level must be investigated for sustainable aquaculture which will not only reduce the running cost but will prevent water quality from deterioration (Abbas and Siddiqui, 2009). Although, some studies have been conducted on the nutrient requirements of demersal fish (Catacutan et al., 2001; Abbas and Siddiqui, 2003; Catacutan and Pagador, 2004; Mourente and Bell, 2006). However, no information about ration level and feeding frequency of black fin sea bream, A. berda is available. Therefore, present study was planned to determine the optimum ration size and feeding frequency of A. berda juvenile growing from 23.5 g to 125.8 g for best growth.

 

MATERIALS AND METHODS

 

Experimental feed preparation

The experimental feed (42% protein and 20% lipid) was prepared from the available ingredients containing fish meal as source of protein, and tapioca was used as source of carbohydrate. Cod liver oil was used as source of lipid for energy. Minerals and vitamins were also added to the experimental diets. All components were weighed, grounded and mixed mechanically to realize homogeneity of ingredients. Water (150 ml/kg) was added to the mixture and was remixed. Thus, soft dough was pelleted by 2mm die. These pellets were then dried under shade for 10 h and stored for further use.

Fish juvenile collection and management

Sea bream juveniles were collected from Sonari coast located towards west from Karachi, and they were acclimatized for 15 days prior to start the experiment. After acclimatization, they were randomly distributed into the glass tanks (150 liter) 10 fish in each. All these tanks were supplied with sand-filtered sea water which was aerated continuously. All the fish were placed in similar photoperiod of 12L:12D. Feed was supplied by hand and uneaten feed was collected after 2 h. The water was cleaned daily by siphoning. Fish length and weight were noted fortnightly.

Experimental procedure and feeding trial

In order to determine the effect of ration level and feeding frequency on growth, two separate trials were conducted in the laboratory of Aquaculture, CEMB, University of Karachi. In Experiment I, 10 fish (mean weight 23.5±0.6 g) were stocked in rectangular tanks in three replication. Fish were randomly distributed in rectangular tanks, and were individually weighed. Six levels (2.0, 2.5, 3.0, 3.5, 4.0 and 4.5% of their body weight per day, BW d−1) of the experimental diet were fed to the fish three times a day in each replication for 75 days.

In Experiment II, juvenile sea bream were treated in the same experimental facility as mentioned in Experiment I. Sea bream juvenile (mean weight 23.5±0.6 g) were randomly distributed in rectangular tanks, 10 fish in each tank with three replication. Feed were supplied with 2.5 BW d-1 with four feeding frequency (once daily at 0800 h, twice daily at 0800 and 1800h, three times daily at 0800, 1300 and 1800 h, and four times daily 0800, 1120, 1440 and 1800 h) for 75 days. The length and weight of fish were noted fortnightly. Uneaten diet was removed by siphoning and thus feed intake was recorded.

Chemical analysis and measurement

At the end of the experiment, three fishes from each tank were killed and then dissected to calculate the weight of liver and viscera for determining the hepatosomatic index (HSI) and viscerosomatic index (VSI). Three fishes were also collected and killed for carcass analysis. Crude lipid (CL), moisture and crude protein (CP) were determined by using the procedure of Association of Official Analytical chemist (AOAC, 2000). The moisture was estimated at 105°C for 24 h with the help of an oven (Labostar-LG122 Tabia Espec, Osaka, Japan). Crude lipid was estimated by chloroform/ methanol (2:1v/v) extraction procedure (Folch et al., 1957). Crude protein was determined by using Kjeldahl method (N×6.25) using automatic Kjeldahl system (Buchi 430/323). Ash was calculated by burning in a muffle furnace. Energy in each treatment was determined with the help of automatic bomb-calorimeter (Parr Instruments, model1265, Moline IL, USA). The data of these parameters were taken as mean of three replicates.

Statistical analysis

The experimental data was analyzed by one way analysis of variance (ANOVA) to determine biological and chemical indices of fish. Difference among means was calculated by 5% probability levels addressing Duncan’s multiple range tests (Zar, 1996). Optimum ration level and feeding frequency for maximum growth of black fin sea bream were estimated by the maximum percent weight gain of initial weight. The weight gain (WG), protein efficiency ratio (PER), specific growth rate (SGR), feed conversion ratio (FCR), feed intake (FI) and condition factor (CF) hepatosomatic index (HSI) and viscerosomatic index (VSI) were determined by the following formulae:

 

CF = 100 × weight / length3.

SGR = 100 × (ln final weight – in initial weight/period).

FCR = diet given / weight gain).

HSI = weight of liver (g)/empty fish weight (g) ×100.

PER = wet weight gain / N×6.25 intake.

VSI = 100 × [wet weight of visceral organs and associated fat tissue (g) / wet body weight (g)].

 

Table I.- Feed formulation and chemical analysis of test diet.

Ingredients1	g 100 g-1 diet (dry)
Fish meal	37.5
Tapioca flour	13.6
Lupine seed meal	6.8
Corn gluten meal	7.5
Wheat flour	12
Vitamin-mineral premix	2.6
Cod liver oil	20
Proximate composition2
Moisture	7.6±0.5
Crude protein3	42.1±2.3
Crude lipid	20.0±1.0
Crude fiber	3.4±0.7
Ash	13.6±0.9
Carbohydrates4	20.9±1.6
Energy (kJ/g)	25.2±1.9

1Rahim et al., 2015. 2Dry matter (%): number of samples = 5. 3Measured as N × 6.25. 4Carbohydrates = 100 – (%protein + %fat + %ash + %fiber).

 

Table II.- The weight gain, specific growth rate, feed conversion ratio, protein efficiency ratio, feed intake of juvenile A. berda fed at different feeding levels in rectangular tanks (Experiment I).

Parameter	Ration level (% BW d−1)
	2.0	2.5	3.0	3.5	4.0	4.5
Final weight	85.2±0.7a	125.5±0.4a	125.8±0.9b	125.6±2.4b	125.8±1.4b	125.9 ±3.8b
WG% 1	262.55±0.5a	434.04±0.8a	435.31±0.4b	434.4±0.6b	435.31±0.5b	435.7±0.4b
SGR2	1.71±0.4a	2.23±0.3b	2.23±0.5b	2.23±0.3b	2.23±0.4b	2.23±0.3b
FCR3	0.4±0.01c	0.20±0.02a	0.27±0.01b	0.27±0.02b	0.27±0.03b	0.27±0.02b
PER4	1.2±0.2a	1.3±0.3b	1.3±0.3b	1.5±0.2c	1.4±0.3bc	1.5±0.3c
FI5	35.1±1.3a	35.3±1.3a	35.2±1.4a	35.2±1.3a	35.1±1.3a	35.1±1.2a
CF6	2.6±0.1a	3.1±0.2a	3.1±0.1a	3.2±0.2a	3.1±0.1a	3.1±0.2a
HSI7	1.2±0.2a	1.4±0.1b	1.4±0.1b	1.4±0.2b	1.4±0.2b	1.4±0.2b
VSI8	6.1±0.1a	6.9±0.1b	7.1±.02b	7.5±0.1bc	7.7±0.2bc	7.7±0.1bc
Survival	100	100	100	100	100	100

Similar superscripts show no significant (P>0.05) difference among treatments. 1WG, % of initial weight =100 × [final weight – initial weight / initial weight. 2CF =100 × weight / length3. 3FI = diet given as % body weight – remaining diet pellets. 4SGR =100 × ln final weight – ln initial weight / period. 5FCR = diet given / WG). 6HSI = wet of liver (g) / empty fish weight (g) ×100: total of initial was 1.24%. 7PER = wet weight gain / N×6.25 intake. 8VSI = 100 × [wet weight of visceral organs and associated fat tissue (g) / wet body weight (g)].

 

RESULTS

 

Water quality

During the study period, water temperature ranged from 27+0.5°C to 28+0.4°C. Salinity was maintained at 20.2±0.7%. Dissolved oxygen (DO) remained 6.8 ml l−1to 7 ml l−1and pH was found to be 6.9 to 7.7. Ammonia (NH4-N) and nitrites (NO2-N) were not more than 0.1±0.008 ml l−1 and nitrates were less than 0.02 ml l−1.

Chemical composition of the experimental diet

The experimental feed was analyzed for achieving desired amounts of protein and lipid. The chemical composition showed that feed contained approximately 42.1% protein, 20.0% lipid, 13.6 % ash, 20.9% carbohydrate, 7.6% moisture and 25.2kJ/g energy (Table I).

Effects of ration size

No disease was noted during the entire experimental duration and thus no mortality was observed. Weight gain (WG) increased considerably (P<0.05) with increasing feeding level up to 2.5% BW d−1 (Table II). Best specific growth rate (SGR) was also noted in fish fed with 2.5% BW d−1. But no further increase was observed when feeding level was increased away from 2.5% BW d−1to 4.5BW d−1. Feed conversion ratio (FCR) shows considerable best value for fish fed with 2.5% BW d−1 and no enhancement were found when it was increased up to 4.5% BW d−1. Protein efficiency ratio (PER) increased slightly with increase in ration level from 2 to 3.5 BW d−1, but further increase in ration level showed no effect on the PER ratio. The proportion of moisture, protein and ash contents of fish whole body were not significantly (P>0.05) affected by feeding levels (Table III). Hepatosomatic index (HSI) viscerosomatic index (VSI), and condition factor (CF) of fish fed 2.0 BW d−1was considerably less than the 2.5, 3.0, 4.0 and4.5% BW d−1 (Table III). Feed intake was not influenced by dietary treatments.

 

Table III.- Whole body composition of A. berda fed with diets of different feeding frequency for 75 days. (Experiment I).

Parameter	Feeding frequency (number of meals d-1)
	1.0	2.0	3.0	4.0
Moisture	70.5±0.07a	70.6±0.21a	70.8±0.08 a	70.7±0.21a
Protein1	17.8±0.14a	18.5±0.27 a	18.9±0.31a	18.8±0.14a
Lipid	10.1±0.14 a	12.1±0.71 a	12.5±0.14a	12.9±0.28 a
Ash	4.4±0.05a	4.3±0.06 a	4.4±0.06a	4.1±0.07a

Similar superscripts show no significant (P>0.05) difference among treatments. Initial body proximate composition was: moisture 71.1%, Protein 17.3%, lipid 9.2% and ash 4.3%. 1Measured as nitrogen × 6.25.

 

Effects of feeding frequency

No pathological symptom and mortality among treatments was noted during this study. Percent WG, SGR, and best FCR were noted in fish fed with three and four times daily. While low percent weight gain and specific growth rate were recorded for the fish fed with once and twice daily (Table IV). PER of fish fed for all treatments was not significantly different. No significant different in feed intake was observed among all treatment. VSI of the fish fed with three and four time a day was found greater than that of one and two times daily. HIS of the fish fed with one time daily was less than that of two, three, and four times. The whole body protein, lipid, moisture, and ash contents of fish were not significantly (P>0.05) affected by feeding frequency (Table V). However, lipid content of fish fed two, three and four times daily was considerably (P<0.05) greater than fish fed one time daily.

 

Table IV.- The weight gain, specific growth rate, feed conversion ratio, feed intake, protein efficiency ratio and condition factor of juvenile A. berda at different feeding frequencies for 75 days (Experiment II).

Parameter	Feeding frequency
	1.0	2.0	3.0	4.0
Final weight	85.5±0.92a	95.5±1.41b	125.8±0.72c	123.6±0.21c
WG%1	263.8±1.28a	306.3±1.21b	435.3±1.41c	425.9±1.21c
SGR2	1.72±0.21a	1.86±0.28a	2.2±0.35b	2.2±0.25b
FCR3	0.13±0.02b	0.11±0.02b	0.08±0.04a	0.082±0.02a
FI4	34.1±0.21a	35.2±0.74b	35.2±0.35b	35.1±0.61b
PER5	1.3±0.04a	1.3±0.51a	1.5±0.08b	1.4±0.04a
CF6	3.1±0.35a	3.2±0.21a	3.1±0.35a	3.1±0.21a
HSI7	1.3±0.14a	1.2±0.35a	1.4±0.07b	1.4±0.21b
VSI8	6.5±0.31a	7.5±0.21b	7.7±0.31b	7.5±0.31b
Survival	100	100	100	100

Similar superscripts show no significant (P>0.05) difference among treatments. 1WG, % of initial weight =100 × [final weight – initial weight / initial weight. 2CF =100 × weight / length3. 3FI = diet given as % body weight – remaining diet pellets. 4SGR = 100 × (ln final weight – ln initial weight / period). 5FCR = diet given / WG). 6HSI = wet of liver (g) / empty fish weight (g) ×100: total of initial was 1.24%. 7PER = wet weight gain / N×6.25 intake. 8VSI = 100 × [wet weight of visceral organs and associated fat tissue (g) / wet body weight (g)].

 

Table V.- Whole body composition of A. berda fed with diets of different feeding frequency for 75 days (Experiment II).

Parameter	Feeding frequency (number of meals d-1)
	1.0	2.0	3.0	4.0
Moisture	70.2±1.03a	71.3±1.11a	70.5±1.04a	70.6±1.22 a
Protein1	17.5±0.14a	18.8±0.25a	18.7±0.21a	18.5±0.34 a
Lipid	10.3±0.12a	12.2±0.51a	12.6±0.12a	12.4±0.17 a
Ash	4.3±0.03a	4.2±0.05a	4.3±0.03a	4.2±0.03 a

Similar superscripts show no significant (P>0.05) difference among treatments. Initial body proximate composition was: moisture 71.2%, protein 17.1%, lipid 9.3% and ash 4.1%. 1Measured as nitrogen × 6.25.

 

DISCUSSION

 

In the present study, best growth performance was noted in juveniles of black fin sea bream (A. berda) fed at 2.5% BW d−1 with feedings of three and four times daily for 75 days. Similar findings were reported by Haruna et al. (2014) and Aderolu et al. (2010). Fish fed at feeding level of 2.5% BW d-1 and feeding frequency of three time daily showed significantly higher WG, SGR and FCR. These results were in agreement with the observations noted for other carnivorous species (Russel et al., 1996; Kayano et al., 1993; Cho et al., 2003; Aderolu et al., 2010; Nekoubin and Sudagar, 2012; Al-Zahrani et al., 2013). However, it is fact that higher feeding level and feeding frequency have adverse effects on water quality and may increase operation cost (Jobling, 2012). Therefore, feeding to fish should be reduced to a minimal level. This minimal level corresponds to 2.5% BW d−1 with three times a day in the present study. Hence optimization of feeding level and feeding frequency is necessary (Abbas et al., 2015). Moreover, feeding level less than 2.5% BWd-1 decreased weight gain, this indicates that sea bream must require 2.5% BWd-1 feeding level for best growth similar to the findings for other fishes with similar feeding behavior (Al-Zahrani et al., 2013; Abbas et al., 2015). Moreover, when feeding was at 2.0% BW d−1 with one and two times a day, fish showed the lowest growth which might have been due to the nutrient requirement for maintenance. It appears that a large proportion of nutrient in the diet was used to maintain life, and only a small proportion was available for growth. Low feeding level and high feeding frequency proved that with excessive ration level, the supplied feed are partially consumed by fish and large amount of feed become useless by dissolved in water deteriorating water quality (Cho et al., 2003; Ribeiro et al., 2012; Abbas et al., 2015). While in moderate ration level and feeding frequency the required amount of feed was available for fish to meet the current feed requirement which enhanced the growth and inhibit water from deterioration parallel with study of many scientist for other demersal fish like sea bass Dicentrarchus labrax (Tsevis et al., 1992). They found best results about growth performance on low feeding level and high feeding frequency.

Feed intake increased with the increase in feeding frequency from one to three times a day, but further increase in feeding frequency from three to four times do not affect feed intake values. This indicates that further increase in feeding frequency dose not enhance the growth but causes wastage of food. Studies conducted on many fishes revealed that feed consumption as well as growth performance generally improved with feeding frequency up to a cretin limit as described by Abbas et al. (2015) which is in line with the present study. It was also noted that excessive feeding of fish caused extra stress on the gut and create gastro problem in digestion (Jobling, 2012). In the present study, PER was not affected by the feeding frequency but ration level slightly affected the PER, increasing slightly with increase in ration level from 1.0% BW d-1 to 3.5% BW d-1 but further increases up to 4.5% BW d-1 have no effects on PER value.

Indices of condition like CF, HIS and VSI play important role for the assessment of nutritional status of fish (Ng et al., 2000; Abbas et al., 2015). In this study, HSI and VSI are not significantly affected by the feeding frequency. These results tally with the findings of Zakes et al. (2006) and Iqbal et al. (2015). In addition, similar results were found by Gines et al. (2004) for gilthead sea bream Sparus aurata. CF was similar among all treatments agreeing with the study of Al- Zahrani et al. (2013). They found that feeding frequency did not affect the condition indices.

Keeping in view the effects of ration size and feeding frequency on whole body composition, crude protein was not significantly affected by the feeding level and feeding frequency. This indicates that all diets can provide the dietary protein for maintenance of the fish body but less feeding level and feeding frequency did not enhance the growth as described by Cho et al. (2003). Positive correlation was found between feeding frequency and whole body lipid in the present study which is in agreement with the study of Jegede and Olorunfemi (2013). He found direct relationship between lipid and feeding frequency. It is fact that optimization of feeding frequency and feeding level not only improve the growth, FCR, and SGR but also decreases the running cost and water deterioration of any culture operation. On the basis of these results, it is suggested that black fin sea bream A. berda (weight 23.5g to 125.5 g) perform best growth on feeding level of 2.5% BW d-1 and feeding frequency of three times a day under the present culture conditions.

 

ACKNOWLEDGEMENTS

 

The senior author is grateful to the HEC for providing facilities and fellowship to complete this work as a part of Ph. D. research.

 

Statement of conflict of interest

Authors have declared no conflict of interest.

 

refereces

 

Abbas, G. and Siddiqui, P.J.A., 2003. Effect of feeding rate on growth, feed conversion and body composition of juvenile mangrove red snapper reared in sea water tanks. Pakistan J. Zool., 35: 151–156.

Abbas, G. and Siddiqui, P.J.A., 2009. Effects of different feeding level on the growth, feed efficiency and body composition of juvenile mangrove red snapper, Lutjanus argentimaculatus (Forsskal 1775). Aquacult. Res., 40: 781–789. https://doi.org/10.1111/j.1365-2109.2008.02161.x

Abbas, G., Siddiqui, P.J.A. and Jamil, K., 2011. The optimal protein requirements of juvenile mangrove redsnapper, Lutjanus argentimaculatus fed isoenergeticdiets. Pakistan J. Zool., 44: 469–480.

Abbas, G., Waryani, B., Ghaffar, A., Rahim, A., Hafeez-ur-Rehman, M. and Aslam, M., 2015. Effect of ration size and feeding frequency on growth, feedutilization, body composition and some haematologicalcharacteristics of juvenile snapper, Lutjanus Johnii (Baloch, 1792). Pakistan J. Zool., 47: 719–730.

Al Zahrani, A.W., Mohamed, A.H., Jr, A.E.S. and Traifalgar, R.F.M., 2013. Effects of feeding rate and frequency on growth and feed utilization efficiency in the camouflage grouper (Epinephelus polyphekadion) fingerlings fed a commercial diet. Eur. J. exp. Biol., 3: 596–601.

Andrews, J.W. and Page, J.W., 1975. The effects of frequency of feeding on culture of cat fish. Trans. Am. Fish. Soc., 104: 317–321. https://doi.org/10.1577/1548-8659(1975)104<317:TEOFOF>2.0.CO;2

Aderolu, A.Z., Seriki, B.M., Apatira, A.L. and Ajaegbo, C.U., 2010. Effects of feeding frequency on growth, feed efficiency and economic viability of rearing African catfish (Clariasgariepinus, Burchell 1822) fingerlings and juveniles. Afri. J. Fd. Sci., 4: 286– 290.

Anonymous, 2012. Hand book of fisheries statistics of Pakistan. A publication of Marine Fisheries Department, Government of Pakistan. Ministry of Food, Agriculture and Cooperatives (Livestock Division), 20: 215 pp.

AOAC, 2000. Official methods of analysis of Association of Official Analytical Chemists. Vol. I. 17th edn. Association of Official Analytical Chemists, Arlington, USA, pp. 684.

Catacutan, M.R. and Pagador, G.E., 2004. Partial replacement of fishmeal by defatted soybean meal in formulated diets for the mangrove red snapper, (Forsskal 1775). Aquacult. Res., 35: 299−306. https://doi.org/10.1111/j.1365-2109.2004.01015.x

Catacutan, M.R., Pagador, G.E. and Teshima, S., 2001. Effect of dietary protein and lipid levels and protein to energy ratios on growth, survival and body composition of the mangrove red snapper, Lutjanus argentimaculatus (Forsskal 1775). Aquacult. Res., 32: 811–818. https://doi.org/10.1046/j.1365-2109.2001.00618.x

Cho, S.H., Lim, Y.S., Lee, J.H., Lee, J.K. and Park, S., 2003. Effects of feeding rate and feeding frequency on survival, growth and body composition of ayu post larvae Plecoglossus altivelis. J. World Aquacult. Soc., 34: 85–91. https://doi.org/10.1111/j.1749-7345.2003.tb00042.x

Folch, A.C., Leed, M. and Sloane-Stanley, G.M., 1957. A simple method for isolation and purification of total lipids from animal tissues. J. biol. Chem., 226: 497–509.

Gines, R., Afonso, J.M., Arguello, A., Zamorano, M.J. and Lopez, J.L., 2004. The effects of long-day photoperiod on growth, body composition and skin colour in immature gilthead sea bream (Sparusaurata L.). Aquacult. Res., 35: 1207–1212. https://doi.org/10.1111/j.1365-2109.2004.01126.x

Haruna, M.A.H., Muhd, I.U., Ahmad, M.K. and Umar, R., 2014. Evaluation of different feeding frequencies on growth performance and feed utilization of Clarias gariepinus (Burchell, 1822) fingerlings. Bayero J. Pure appl. Sci., 7: 142–144.

Iqbal, K.J., Ashraf, M., Qureshi, N.A., Javid, A., Abbas, F., Rehman, M.H., Rasool, F., Khan, N. and Abbas, S., 2015. Optimizing growth potential of Labeo rohita fingerlings fed on different plant origin feeds. Pakistan J. Zool., 47: 31–36.

Jegede, T. and Olorunfemi, O.T., 2013. Effects of feeding frequency on growth and nutrient utilization of Oreochromis niloticus (Linnaeus 1757) fingerlings. Global J. Inc., 13: 0975–5896.

Jobling, M., 2012. Nutrient requirements of fish and shrimp. National Research Council (NRC): The National Academies Press, Washington, D.C., 2011, pp, 376+XVI.

Kayano, Y., Yao, S., Yamamoto, S. and Nakagawa, H., 1993. Effects of feeding frequency on the growth and body constituents of young red spotted grouper, Epinephelus akaara. Aquaculture, 110: 271–278. https://doi.org/10.1016/0044-8486(93)90375-9

Mourente, G. and Bell, J.G., 2006. Partial replacementof dietary fish oil with blends of vegetable oils (rapeseed, linseed and palm oils) in diets for European sea bass (Dicentrarchuslabrax L.) over a long term growth study: Effects on muscle and liver fatty acid composition and effectiveness of a fish oil finishing diet. Comp. Biochem. Physiol., 145: 389−399. https://doi.org/10.1016/j.cbpb.2006.08.012

Nekoubin, H. and Sudagar, M., 2012. Effects of feeding frequency on growth performance and survival rate of grass carp (Ctenopharyngodon idella). World appl. Sci. J., 178: 1001-1004.

Ng, W.K., Lu, K.S., Hashim, R. and Ali, A., 2000. Effects of feeding rate on growth, feed utilization and body composition of a tropical bagrid catfish. Aquacult. Int., 8: 19–29. https://doi.org/10.1023/A:1009216831360

Rahim, A., Abbas, G., Waryani, B., Ghaffar, A., Monwar, M.M., Hafeez-ur-Rehman, M. and Dastagir, G., 2015. Influence of varying dietary lipid levels on growth, feed conversion and chemical composition of meat and liver of the juvenile black fin Sea bream, Acanthopagrus berda (Forsskal 1775). Pakistan J. Zool., 47: 1467-1473.

Russell, N.R., Fish, J.D. and Wootton, R.J., 1996. Feeding and growth of juvenile sea bass: the effect of ration and temperature on growth rate and efficiency. J. Fish Biol., 49: 206–220. https://doi.org/10.1111/j.1095-8649.1996.tb00017.x

Ribeiro, F.D.A.S., Vasquez, L.A., Fernandes, J.B.K. and Sakomura, N.K., 2012. Feeding level and frequency for freshwater angel fish. R. Bras. Zootec., 41: 1550–1554. https://doi.org/10.1590/S1516-35982012000600033

Tsevis, N., Kloudatos, S. and Conides, A., 1992. Food conversion budget in sea bass, Dicentrarchus labrax, fingerlings under two different feeding frequency patterns. Aquaculture, 101: 293–304. https://doi.org/10.1016/0044-8486(92)90032-G

Wang, Y., Kong, L.J., Li, K. and Bureau, D.P., 2007. Effects of feeding frequency and ration level on growth, feedutilization and nitrogen waste output of cuneate drum (Nibea miichthioides) reared in net pens. Aquaculture, 271: 350–356. https://doi.org/10.1016/j.aquaculture.2007.03.022

Zakes, Z., Kowalska, A., Czerniak, S. and Demska-zakes, K., 2006. effect of feeding frequency on growth and size variation in juvenile pike perch, Sander lucioperca. Czech J. Anim. Sci., 51: 85–91.

Zar, J.H., 1996. Biostatistical analysis. Prentice-Hall Inc., New Jersey, pp. 662.