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Effect of Feeding Frequency on Growth Performance, Feed Utilization and Body

Composition of Juvenile Nile Tilapia, Oreochromis niloticus (L.) Reared in Low
Salinity Water

Article in Pakistan journal of zoology · January 2016

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Pakistan J. Zool., vol. 48(1), pp. 171-177, 2016.

Effect of Feeding Frequency on Growth

Performance, Feed Utilization and Body
Composition of Juvenile Nile Tilapia,
Oreochromis niloticus (L.) Reared in
Low Salinity Water
Abdul Malik Daudpota,1 Ghulam Abbas,1,* Illahi Bux Kalhoro,2 Syed Sajjad A. Shah,3
Hameeda Kalhoro,4 Muhammad Hafeez-ur-Rehman5 and Abdul Ghaffar6
1
Centre of Excellence in Marine Biology, University of Karachi, Karachi
2
Department of Anatomy and Histology, Faculty of Animal Husbandry and Veterinary
Science, Sindh Agriculture University, Tando Jam
3
Fish Hatchery Chilya, Thatta, Directorate of Fisheries Sindh (Inland), Hyderabad
4
Department of Fresh Water Biology and Fisheries, University of Sindh, Jamshoro
5
Department of Fisheries and Aquaculture, University of Veterinary and Article Information
Animal Sciences, Lahore Received 28 May 2015
6 Revised 10 July 2015
Department of Life Sciences, Islamia University of Bahawalpur, Bahawalpur Accepted 11 July 2015
Available online 1 January 2016
AB S TR AC T
Authors’ Contributions:
Feeding frequency is well known for the reduction of aquaculture production cost and preventing water quality AMD did the experimental work.
deterioration as the result of excess feeding. The influence of feeding frequency was studied in the present study, SSAS maintained fish stocks. HK
when juvenile Nile tilapia Oreochromis niloticus (initial body weight 1.0 g) were maintained in rearing tanks 35 prepared fish feed. AG analyzed feed
liters each (water temperature 29oC, salinity 14‰, pH 7.7, dissolve oxygen 5.1ml/l and ammonia never exceeded components. MHR analyzed fish
0.1 ml/l) and were fed diet of 35% protein for 42 days. Fish were fed at four frequencies: two, three, four and five meet. GA designed the experiment,
times a day. Each feeding frequency was assigned to three tanks of fish with 10 fish per tank. Results showed that wrote the manuscript with the help
significantly higher weight gain, specific growth rate, feed conversion and protein efficiency ratio were observed at of IBK.
feeding frequency of four to five times daily. Moisture, protein and ash contents of whole body were not affected
by feeding frequency. Lipid content of fish fed four and five times daily was significantly higher than that of the Key words:
fish fed one and two times daily. The condition factor remained consistent at all feeding frequencies and survival Oreochromis niloticus, growth, feed
was 100% throughout the experiment. These results suggest that under similar culture conditions, the optimum conversion, feeding frequency, body
feeding frequency of juvenile Nile tilapia (from initial body weight of 1.0 g to 5.8 g) is four times daily. composition.

INTRODUCTION al., 1993; Cho et al., 2003). Knowledge about the

optimum feeding is important not only for regulating the
feed intake, growth and chemical composition of fish but
T he commercial feasibility of any intensively also for preventing water quality deterioration as a result
of overfeeding (Du et al., 2006; Salama, 2008; Ertan et
cultured fish species depends on market demand and cost
of production. The largest section of the production cost al., 2015). On the other hand, overfeeding of fish can
lies in feed, with protein containing the most expensive overload the stomach and intestine, leading to decreases
component (Pandian, 1989; National Research Council, in digestive efficiency and reductions in feed utilization
1993). However, to develop fish culture at commercial (Jobling 1986; Hung and Lutes, 1987; Storebakken and
level, it is important to establish an appropriate feeding Austreng, 1987; Du et al., 2006). Thus, the diet amount
management strategy that is based on identification of the fed each time, or feeding frequency, may influence diet
daily feeding patterns or rhythms (Phillips et al., 1998; utilization (Sanches and Hayashi, 2001). This is due to
Wang et al., 1998). It is well known that the amount and the fact that diet is directly applied to water and the non-
timing of feeding play an important role in growth and uptaken portion will be dissolved and lixiviated. Feed
feed utilization efficiency (Brett and Groves, 1979; conversion ratio increase and environmental pollution are
Reddy and Katre, 1979; Hung and Lutes, 1987; Hung et the results. Since fish juveniles uptake a high daily diet
ratio to meet their nutritional requirement and thus ingest
adequate amount of diet, and since high feeding
_______________________________
* Corresponding author: [email protected]
frequency results in high daily diet intake ratio and small
0030-9923/2016/0001-0171 $ 8.00/0 amounts of diet per feeding (Sanches and Hayashi, 2001),
Copyright 2016 Zoological Society of Pakistan
172 A.M. DAUDPOTA ET AL.

a higher frequency may be the most adequate. The experimental feeds were then stored at –20C for
Tilapia (Oreochromis spp.) are known as feeding trials.
commercially important food fishes for aquaculture
throughout several regions of the world such as China, Table I.- Formulation and chemical analysis of the
South-east Asia, Africa, USA and Latin experimental diet.
America/Caribbean (Lim and Webster, 2006;
Chowdhury, 2011). According to FAO (2012) global Ingredients (%) g 100 g-1 diet (dry)
tilapia production, which totaled less than 500,000 metric
Fish meal 34.5
tons in the early 1990s, topped 3.5 million metric tons in
Wheat brawn 16.0
2011. In 2012 it increased up to 2.7 percent, and further Rice brawn 11.0
climbed to 3.4 percent in 2013. By 2014, it is expected to Mustered oil cake 13.0
approach around 3.9 million tons. In Pakistan, tilapia is Wheat flour 20.0
known for its good quality meat and it is cheaper as Vitamin-mineral premix1 2.5
compared to other food fishes. It can play an important Fish oil 3.0
role in aquaculture development in the country. Few
feeding trials indicated that Nile tilapia (Oreochromis Proximate composition2
niloticus) has potential for aquaculture in Pakistan Moisture 7.0±0.5
Crude protein3 34.6±0.5
because of its hardiness and rapid growth (Jamil et al.,
Crude lipid 5.8±0.6
2004; Khan et al., 2014; Chughtai et al., 2015). Crude fiber 5.9±0.4
Although some work has been done on the feeding Ash 7.0±0.7
patterns of Nile tilapia (Macintosh and Little, 1995; Tung NFE4 46.7±0.3
and Shau, 1991; Vera Cruze and Mair, 1994; Sanches and Energy (kJg-1) 20.2±0.6
Hayashi, 2001), data regarding feeding frequency for this P/E (mg crude protein kJ-1) 17.1±0.5
species in Pakistani waters is scarce. It was in view of
1
this paucity of information that the present study was Vitamin and mineral mixture contained the following
carried out to investigate the influence of feeding ingredients (g 100 g−1 diet): Ascorbic acid (vit C), 15.2;
frequency on the growth, feed efficiency and body thiamin HCl (vit B6), 1.1; inositol, 39.5; calcium, 1.25; zinc,
1.0; retinol (vit A), 1.5; phosphorus, 3.5; choline chloride, 3.5;
composition of juvenile Nile tilapia for a period of 42 magnesium, 2.0; copper, 1.0; pyridoxine (vit B6), 1.3;
days. In this study, a low-cost feed was formulated using phospholipids, 3.5; α-tocopherol acetate (vit E), 5.5; folic acid,
locally-available ingredients and was supplied to the fish 0.4; cholecalciferol (vit D3), 7.5; cyanocobalamine (vit B12),
at different feeding frequencies for 42 days. 0.006; riboflavin (vit B2), 1.5; menadione sodium bisulphite
(vit K3), 0.03; manganese, 2.0; iodine, 2.0; sodium, 1.0; iron,
MATERIALS AND METHODS 1.0; nicotinic acid, 4.3; biotin, 0.35.
2
Dry matter basis (%): mean ± SE, number of determination =3.
3
Experimental diet Measured as nitrogen × 6.25.
4
Fish feed (20.2 kJ g-1 digestible energy) was Nitrogen-free extract = 100 – (% protein + % fat + % ash + %
fiber).
formulated on dry matter basis (g 100-1) in one batch to
supply calculated protein levels of 35% with fishmeal
providing the majority of dietary protein (Table I). A Experimental design
mixture of minerals and vitamins were added to the Juveniles of Nile tilapia, Oreochromis niloticus
ingredients (rice bran, wheat bran, mustered oil cake and (mean weight 1±0.01g and mean length 3.8±0.02 cm)
wheat flour) of diets. All these ingredients were collected from Government Fish Hatchery, Chilya,
purchased from the local markets and were ground to Thatta, Sindh were held in seawater for fifteen days
500µm and mechanically mixed for15 min to ensure before starting the experiment. After the acclimatization
homogeneity. Fish oil was added and then mixed again phase, fish were randomly distributed in twelve (12)
for 15 minutes. Water (250 mL kg-1 dry ingredients plastic experimental indoor tanks (10 fish per tank). The
mixture) was added and mixed for another 15 minutes to water carrying capacity of each tank was 35 liters.
attain a consistency appropriate for pelleting. The wet Aeration was provided by aerators throughout the entire
mash was pelleted with a California Laboratory Pellet experiment which lasted 42 days. Fish were subjected to
Mill (model CL-type 3, California Pellet Mill Company, a natural photoperiod and all tanks had similar light
San Francisco, CA, USA.) using a 2-mm die. No heating conditions. Physico-chemical parameters i.e.,
or steam was used in the pelleting process and the wet temperature, salinity, pH and dissolved oxygen were
pellets were air-dried at room temperature for 20 hours. monitored daily.
 FEEDING FREQUENCY OF NILE TILAPIA JUVENILES 173

Feeding protocol weight gain (expressed as the percent of initial body

Experimental diet containing 35% protein weight at the end of the experiment), specific growth rate
concentration was tested to find out the optimum feeding (SGR) (ln final body weight – ln initial body weight/time,
frequency of Nile Tilapia (Oreochromis niloticus). expressed as % per day), feed conversion ratio (FCR)
During this experiment, diet was supplied to the (feed fed/wet weight gain), protein efficiency ratio (PER)
experimental juveniles with the frequency of two, three, (wet weight gain/protein intake).
four and five times per day. Fish were hand-fed on daily
ration of 3% wet body weight per day for 42 days. The Statistical analysis
daily feed supplied was recorded and uneaten feed was The data on fish growth, feed utilization efficiency
collected two hours after the start of feeding. The amount and whole fish body constituents were subjected to one-
of food to be provided being adjusted following weekly way analyses of variance (ANOVAs) to determine
sampling for the determination of gain in weight and whether there was a significant difference (P<0.05)
length per treatment which lasted 42 days. Each tank was among fish fed at different protein levels. Differences
completely drained and thoroughly scrubbed on the day between means were assessed at the 5% probability level
of sampling. using Duncan’s multiple range test, as described by
Robbins et al., (1979) and Steel and Torrie (1980). The
Measurement and analysis data are presented as mean±SE of the replicate groups.
At the end of study five fish were removed from
each tank, killed and pooled for whole body composition RESULTS
analysis. Fish whole-body samples were taken out of the Water quality
–20 C cold store and thawed at room temperature using The water temperature was maintained at
a fan. Subsequently, all these samples were homogenized, 29±0.5°C (mean±SD). Salinity was 14.0±0.7‰ and pH
dried and then ground into a powder before chemical ranged from 7.6 to 7.9 with a mean of 7.7±0.05
composition analysis. throughout the study period. Dissolve oxygen was
At the end of the experiment, three replicate 5.1±0.1 ml/l. Ammonia never exceeded 0.1±0.004 ml/l.
samples with10 fish per replicate were taken and kept
frozen at –20 C for subsequent analysis of the fish whole Growth performance
body composition. Growth performance of Nile tilapia juveniles was
The moisture, protein, lipid and ash contents of significantly affected by feeding frequency (Table II).
experimental diets and samples were analyzed according Body weight gain and SGR of the fish fed four and five
to the standard methods (Association of Official times daily were significantly (P<0.05) higher than of
Analytical Chemists 2000). Moisture was determined by those fed the two and three times daily. Weight gain and
drying in an oven (Labostar-LG 122, Tabai Espec, Osaka, SGR tended to plateau at around 480.0 g and 4.19% day-1
Japan) at 105 C for 24 h; ash by burning in a muffle respectively.
furnace (Isuzu Seisakusho, Tokyo, Japan) at 550C for 18
h; crude protein by the Kjeldahl method (N × 6.25) using Feed conversion and condition indices
an automatic Kjeldahl System (Buchi 430/323, Flawil, Feed intake, expressed on a dry matter basis,
Switzerland); crude fiber by acid detergent fiber analysis; increased slightly with an increase in feeding frequency.
and crude lipid by the chloroform/ methanol (2:1, v/v) Fish fed the four and five times daily showed
extraction procedure (Folch et al., 1957). The significantly higher (P<0.05) feed intake than the other
carbohydrate content was calculated by subtracting the groups (Table II). The same trend was observed in feed
content of lipids, total protein and ash from the dry conversion ratio (FCR) and PER. There were no
weight, and gross energy estimation was made using an significant differences in condition factor (CF) between
automatic bomb-calorimeter (Parr Instrument, all the groups where survival remained 100%.
model1265, Moline, IL, USA). All chemical analyses
were performed in triplicate and averaged. Body composition
The moisture, protein and ash contents of fish
Calculation of growth parameters whole body were not significantly (P>0.05) affected by
At the end of the experiment, all fish from each feeding frequency (Table III). The lipid content of fish
tank were individually weighed and their total length was fed four and five times daily was significantly (P<0.05)
measured for calculation of the condition factor [CF = higher than that of fish fed two and three times daily. The
(100 × body weight in g)/ (TL in cm) 3]. Growth and feed lipid contents of whole body increased significantly
efficiency were monitored in terms of the final weight, (P<0.05) with increasing feeding frequency (Table III).
174 A.M. DAUDPOTA ET AL.

Table II.- The growth rate and feed utilization of juvenile Nile tilapia fed at different frequencies for 42 days.

Feeding frequency
Parameters 2 3 4 5

Final weight (g) 3.5±0.1a 4.6±0.3b 5.8±0.5c 5.9±0.4c

Weight gain, % of initial weight1 250.0±1.6a 360.0±2.5b 480.0±2.2c 490.0±1.0c
Specific growth rate2 2.98±0.01a 3.63±0.03b 4.19±0.03c 4.23±0.01c
Feed intake3 (g fish-1) 10.5.±1.3a 13.3±1.5b 16.6±1.1c 17.0±1.0c
Feed conversion ratio4 4.2±0.2a 3.69±0.1a 3.46±0.5b 3.47±0.2b
Protein efficiency ratio5 1.28±0.04a 1.48±0.01a 2.13±0.02b 2.39±0.01b
Condition factor6 2.5±0.03 a 2.6±0.04 a 2.7±0.03 a 2.6±0.04 a
Survival (%) 100 100 100 100

Values (means±SE, n = 3 and each n consists of 10 fish per replicate) in the same row with different superscripts are significantly
different (P>0.05). Initial body weight and length of the fish was 1±0.05 g and 3.5 cm ± 0.01 respectively.
1
Weight gain, % of initial weight = 100 × [final body weight – initial body weight / initial body weight].
2
Specific growth rate = 100 × [ln final body weight – ln initial body weight / time in days].
3
Feed intake = total feed fed as % body weight – total uneaten feed.
4
Feed conversion ratio = total feed fed (g) / total wet weight gain (g).
5
Protein efficiency ratio = wet weight gain / protein (N × 6.25) intake.
6
Condition factor (CF) = 100 × (weight / length3).

Table III.- Whole body composition (% dry weight basis) of Nile tilapia juvenile fed at different frequencies for 42 days.

Feeding frequency
Parameters 2 3 4 5

Moisture 71.2±1.9a 71.0±1.7a 71.3±1.5a 71.2±1.5a

Protein 52.8±2.1a 52.9±1.5a 53.1±1.2a 53.0±1.4a
Lipid 33.6±1.8a 33.5±1.4a 35.1±1.3b 35.4±1.5b
Ash 12.2±1.5a 12.4±1.3a 12.6±1.1a 12.4±2.0a

Values (mean±SE, n =3 and each n consists of 10 fish per replicate) in the same row with different superscripts are significantly
different (P>0.05). Chemical composition of initial body was: moisture 70.0%, protein 51.5%, lipid 33.0% and ash 11.2%

DISCUSSION for growth. The finding that higher growth of Nile tilapia
accompanied with higher food conversion similar to the
In the present study, juveniles of Nile tilapia were findings in other fishes, such as sturgeon Acipenser
shown to require feeding frequency of four times or five transmontanus (Hung and Lutes, 1987), rainbow trout
times daily for the highest growth performance, Oncorhynchus mykiss (Storebakken and Austreng, 1987),
associated with significantly higher weight gain and striped bass Morone saxatilis (Hung et al., 1993), tilapia
better feed conversion. Since growth and feed conversion Tilapia aureus (Papoutsoglou and Voutsinos, 1988),
are influenced by feedings and higher feedings are clownfish Amphiprion percula (Johnston et al., 2003),
counterproductive (Brett and Groves, 1979; Reddy and bagrid catfish Mystus nemurus (Ng et al., 2000) and grass
Katre, 1979; Hung and Lutes, 1987; Hung et al., 1993; carp Ctenopharyngodon idella (Du et al., 2006).
Ng et al., 2000). Therefore, feeding to fish should be Regardless of the existence of these fish species and
reduced to a minimal level. This minimal level categories, many other authors reported that feeding of
corresponds to four times a day in the present study. four times a day is the best for tilapia fry (Abucay and
Feeding above four times daily was over the satiety level Mair, 1977; Jobling, 1983; Macintosh and De Silva,
of the fish and some food waste might occur. Moreover, 1984; Poma and Green, 1990; Tung and Shiau, 1991;
when feeding was with two and three times a day, fish Chambers, 1984; Vera Cruz and Mair 1994; Ridha and
showed the lowest growth which might have been due to Lone, 1995; Cui et al., 1994). These observations are in
the nutrient requirement for maintenance. It appears that conformity with the results of the present study.
a large proportion of nutrient in the diet was used to The proper feeding frequency for maximum
maintain life, and only a small proportion was available growth of fish may differ by fish species, fish size,
 FEEDING FREQUENCY OF NILE TILAPIA JUVENILES 175

dietary protein and energy contents (Minton, 1978; Wang preferred energy reserve for deposition or mobilization
et al., 1998; Company et al., 1999; Lee et al., 2000). over protein in juvenile tilapia which is long-lasting by
Generally, fish fry one fed smaller meals with greater the lower lipid contents of fish whole body were found at
frequency while once per day is usually sufficient for lower feedings.
broodstock and older fish. Usually, the quantity of feed The findings of this study suggest that the
required for optimal growth diminishes as fish age optimum feeding frequency of juvenile Nile tilapia, O.
(Schnai et al., 2005). For instance, Pullin and Lowe- niloticus growing from 1 g to 5.8 g was four meals a day.
McConnell (1982) recommended feeding tilapia, O. At this optimum feeding, improved weight gain, greater
niloticus 12 times daily when their weight is 20 g but food intake, higher feed conversion and superior protein
only twice daily when they weigh 200 g. Rainbow trout retention efficiency are factors apparently responsible for
(0.3 g) grows best when fed eight times per day, but the faster growth rate and lipid reserves in juvenile Nile
similar results were obtained when 15 g fish were fed tilapia under the present experimental conditions.
three times per day (Piper, 1982).
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