Effects of Feeding Frequency on Growth Performance and Survival Rate of Zebra Fish (Danio rerio)

Abstract

This study (at 60 days) was carried out to evaluate the effect of different feeding frequency on growth performance and survival rate of zebrafish (Danio rerio) larvae. Five groups of zebrafish larvae (average weight 0.15 g) were fed either five meals per day (T1), four meals per day (T2), two meals per day (T3), one meal per day (T4) and every other day (T5) and were compared with each other. The Zebrafish larvae in experimental treatments were fed with 5 percent body weight. A higher final body weight (FBW) and specific growth rate (SGR) was obtained in treatment T1 and T2 and were significantly higher than other groups (P<0.05), but these treatments (T1 and T2) were not significantly different when compared with each other (P>0.05). The best feed conversion ratio (FCR) was observed in T2 (four meals per day). However T1 and T2 were not significantly different (P>0.05) in growth performance but T2 showed better result. Survival rate among the treatments were not significantly different (P>0.05).

Full text

Advanced Journal of Agricultural Research
Vol. 1(001), pp. 007-010, May, 2013
©2013 Advanced Journals
http://www.advancedjournals.org/AJAR
Short Communication
Effects of Feeding Frequency on Growth Performance and Survival Rate
of Zebra Fish (Danio rerio)
Hamed NEKOUBIN
1
* Golin RAKHSHANIPOUR
2
, Shima HATEFI
1
, Mohammad SUDAGAR
1
and Salar
MONTAJAMI
1
1
Department of Fishery, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
2
Department of Agricultural Extension and Education, Islamic Azad University of Birjand, Iran.
*Corresponding author. E-mail: nekoubin.hs@gmail.com
Accepted 26
th
April 2013
This study (at 60 days) was carried out to evaluate the effect of different feeding frequency on growth
performance and survival rate of zebrafish (Danio rerio) larvae. Five groups of zebrafish larvae (average
weight 0.15 g) were fed either five meals per day (T1), four meals per day (T2), two meals per day (T3), one
meal per day (T4) and every other day (T5) and were compared with each other. The Zebrafish larvae in
experimental treatments were fed with 5 percent body weight. A higher final body weight (FBW) and
specific growth rate (SGR) was obtained in treatment T1 and T2 and were significantly higher than other
groups (P<0.05), but these treatments (T1 and T2) were not significantly different when compared with
each other (P>0.05). The best feed conversion ratio (FCR) was observed in T2 (four meals per day).
However T1 and T2 were not significantly different (P>0.05) in growth performance but T2 showed better
result. Survival rate among the treatments were not significantly different (P>0.05).
Key words: Zebrafish (Danio rerio), growth performance, survival rate, feeding frequency.
INTRODUCTION
Over the past twenty years, the zebrafish (Danio rerio)
has emerged as an excellent vertebrate model for
studying genetics and development (Fishman, 2001) and
more recently, for the screening of therapeutic drugs
(Penberthy et al., 2002; Lamason et al., 2005). The
zebrafish and human genomes have been shown to
share extensive conserved synthetic fragments and are
therefore increasingly seen as a powerful and highly
amenable model system for many human and animal
diseases with complete genome, available. In addition,
many zebrafish genes and their human homologue
display structural and functional similarities (Fishman,
2001).
Ornamental fish farming is an important primary
industry. Ornamental fishes are often referred as living
jewels due to their color, shape and behavior. They are
peaceful, generally tiny, attractively colored and could
be accommodated in confined space (Kasiri et al., 2011;
Lim and Wong, 1997). Modern ornamental fish culture
and breeding operations, have become vertically and

008 Adv. J. Agric. Res.
horizontally intensified, necessitating a continuous
supply of nutritionally balanced, cost effective feed
(Kasiri et al., 2011; Mandal et al., 2010).
Since diet cost represents 30-70% of the total
operating cost of an aquaculture enterprise (Webster et
al., 2001), determination of the appropriate daily feeding
rate and frequency required to give optimal growth and
feed efficiency could reduce the amount of diet fed,
decrease the amount of time involved in feeding, and
then increase profits. Results of feeding suggest that
feeding rate or frequency may significantly influence
growth rate and body composition. Therefore, in
different fish species cultured under various
environmental and husbandry conditions, more effort is
needed to calculate the optimum feeding rates and
frequencies (Andrews and Pages, 1975; De Silva and
Anderson, 1995). Many authors studied the effect of
feeding frequency on food intake and growth on edible
fishes (Jobling, 1983; Bascinar et al., 2011) but little
attention has been paid to the impact of feeding
frequency on growth and reproductive performance in
ornamental fishes (James and Sampath, 2003).
The present study was aimed at determining optimum
daily feeding frequencies of zebrafish.
MATERIALS AND METHODS
Fish
Larvae of zebrafish with initial weight, 0.15±0.1 g were
obtained from an Institute of commercial supplier, the
Ornamental Fish Hatchery in Gorgan, Iran. They were
kept in glass aquariums (each with a dimension of
30×40×60 centimeters). This experiment was conducted
in a completely randomized design with five treatments
and three replicates per treatment for a total of thirty
Larvae of zebrafish. The density of fish larvae per
aquarium were 10 fishes. The fish were weighed
individually at the beginning, during and at the end of
the experiment. Water quality parameters of input
water to rearing system were monitored each week
throughout the experimental. The water temperature
was 19.46±1.23°C, pH was 7.85±0.26 and water oxygen
level was maintained above 7.65 ± 0.55 MG L
-1
during
the experiment by an electrical air pump (using a single
filtration unit).
Feed analysis
Nutrient compositions of experimental diets (Biomar)
Table 1. Nutrient composition of experimental diets.
.
Ingredients %
Protein 54
Lipid 18
Fiber 1.5
Ash 10
Vitamin 2
are given in Table 1. Proximate composition of diets was
carried out using the Association of Analytical Chemists
(AOAC, 2000) methods. Protein was determined by
measuring nitrogen (N×6.25) using the Kjeldahl method;
Crude fat was determined using petroleum ether (40–60
Bp) extraction method with Soxhlet apparatus and ash
by combustion at 550°C.
Feeding frequency
In experiments, fishes were fed by commercial extruder
diet (Biomar) to satiation, the fishes con-sumed all of the
food in primal five minutes. Triplicates of five feeding
schedules were tested: (T1) (6 A.M., 11 A.M., 16 P.M., 21
P.M. and 12 A.M.), four meals per day (T2) (6 A.M., 12
P.M., 18 P.M., and 12 A.M.), two meal per day (T3) (6
A.M. and 18 P.M.), one meal per day (T4) (6 A.M.) and
every other day (T5) (6 A.M.) for 60 days.
Determination of growth parameters
Growth parameters were calculated as follows:
Body Weight Gain (BWG) = final fish weight (G) – initial
fish weight (G) (Tacon, 1990)
Specific growth rate (SGR) = (Ln W
t
- Ln W
0
) × 100 t
-1
(Hevroy et al., 2005)
Feed conversion ratio (FCR) = P/(W
t
– W
0
) (Shalaby et al.,
2006)
Daily growth rate (DGR) = {100 × (Final weight (g) –
Initial Weight (G))/ (days× Initial weight (G))} (De Silva
and Anderson, 1995)
Survival rate = (N
t
× 100 N
0
-1
) (Ai et al., 2006)

Nekoubin et al. 009
Table 2. Growth parameters and survival rate of zebrafish (Danio rerio) larvae in experimental treatments (trial 1-5).
Growth Indices T1 T5 T4 T3 T2
Initial weight (g) 0.15±0. 01 0.15±0. 01 0.15±0. 01 0.15±0. 01 0.15±0. 01
Final body weight (g) 0.231±0.1
a
0.234±0.1
a
0.191±0.0
d
0.201±0.1
c
0.216±0.1
b
Body weight Gain (g) 0.76±0.2
a
0.83±0.0
a
0.4±0.0
d
0.53±0.1
c
0.67±0.1
b
Specific growth rate for weight (% BWday
1
) 0.69±0.04
a
0.74±0.04
a
0.04±0.0
d
0.51±0.05
c
0.61±0.05
b
Feed Conversion Ratio (%) 4.47±0.11
c
4.4±0.1
c
5.35±0.15
a
5.3±0.07
a
5.1±0.1
b
Feed Conversion efficiency (%) 0.22±0.1
a
0.23±0.1
a
0.19±0.01
b
0.19±0.0
b
0.2±0.0
b
Daily growth rate (DGR) 0.85±0.06
a
0.92±0.06
a
0.44±0.0
d
0.59±0.05
c
0.74±0.06
b
Survival rate (%) 98.79±2.18
a
98.79±2.18
a
94.28±4.28
a
96.18±2.18
a
96.18±2.18
a
Groups with different alphabetic superscripts differ significantly at P<0.05 (ANOVA).
Statistical analysis
In order to determine significant differences, results
were analyzed by one-way Analysis of variance (ANOVA)
and Duncan’s multiple range tests were used to analyze
the significance of the difference among the means of
treatments by using the SPSS program.
RESULTS
In the present study, the zebrafish larvae were subjected
to different daily feeding frequencies at five feeding
rates. The results clearly showed that increasing the
feeding frequency had beneficial effects on the growth
parameters on Zebrafish larvae. The feeding and growth
parameters of Zebrafish larvae are presented in Table 2.
Maximum Body Weight Gain (BWG) was observed in T2
(0.83±0.0) and the lowest BWG was observed in T5
(0.4±0.0). Also specific growth rate (SGR) values of
treatments T2 (0.74±0.04) and T1 (0.69±0.04) were
significantly higher than other groups (P<0.05) and
lowest SGR was observed in T5 (0.04±0.0). Daily growth
rate (DGR) was also improved with the feeding
frequency, and the difference was significant among
treatments (P<0.05). Maximum DGR was observed in T2
(0.92±0.06) and T1 (0.85±0.06) which was significantly
different compare to other groups (P<0.05), however
DGR in T2 was higher than T1, but was not significantly
different to T1 (P>0.05).
The growth parameters were significantly affected by
feeding frequency (P<0.05). Also the food conversion
ratio (FCR) by increase feeding frequency was
significantly decreased in comparison to those of
other groups (P<0.05) and the lowest FCR was observed
in T2 (4.4±0.1) and T1 (4.47±0.11) and had significantly
different (P<0.05) to other groups.
DISCUSSION
Optimum feeding frequency, for maximum growth of
fish, may vary with species and size of fish, culture
conditions, food quality, amount of feed provided and
water temperature (Lee et al., 2000; Kestemont and
Baras, 2001). Studies conducted on other fish species
have shown that feed consumption and growth generally
increased with feeding frequency up to a given limit
which is in agreement with our findings (Kasiri et al.,
2011; Bascinar et al., 2011; Wang et al., 1998). In the
present study, zabrafish fed with four meals per day (T2)
were better than five meals per day (T1) and two meals
per day (T3). Similar research was reported by Grayton
and Beamish (1977), who fed rainbow trout fry and
found out that three meals a day were better than one
and six meals per day. Kasiri et al. (2011) also found out
from their research that feeding angelfish juvenile four
meals a day was better than two or one meal per day.
Feeding frequency in this study did not have effect on
the survival rate of zebrafish (P>0.05) however the
results in T1 and T2 (98.79±2.18) were higher than other
treatments. Similar results were reported by Goldan et
al. (1997) and Aydin et al., (2011) who found out that
feeding frequencies did not significantly affect the
survival rates of sea bream (Sparus aurata) and juvenile
black sea turbot (Psetta maxima).
Ornamental fish farmers incur a higher cost when
cultured fish are fed five times daily compared with four
daily because more time and labor are required and
more feed is likely to be wasted. This will also increase

010 Adv. J. Agric. Res.
the production costs of fish. Based on food consumption
data and growth performance, the highest SGR and
lowest FCR were obtained with fish fed four times a day,
in the present study. It appears that, feeding four times a
day to satiation may be accepted as sufficient for on
growing of zebrafish under the conditions of this
experiment.
ACKNOWLEDGEMENTS
The authors are grateful to M. Hosseinzadeh for the
assistance with the field studies.
REFERENCES
Ai Q, Mai K, Tan B (2006). Replacement of fish meal by
meat and bone meal in diets for large Yellow croaker
(Pseudosciaena crocea). Aquaculture, 260: 255-263.
Andrews JW, Pages JW (1975). The effects of frequency
of feeding on culture of catfish, Trans. Am. Fish Soc.,
104: 317- 321.
AOAC (2000). Oficial methods of analysis. Association of
oficial analytical chemist. EUA, 2000.
Aydın I, Kucuk E, Şahin E, Kolotoglu L (2011). Effect of
feeding frequency and feeding rate on growth
performance of juvenile black sea turbot (Psetta
maxima), J. Fish., 5(1): 35-42.
Bascinar N, Cakmak E, Cavdar Y, Aksungur N (2007). The
effect of feeding frequency on growth performance
and feed conversion rate of Black sea trout (Salmo
trutta labrax Pallas, 1811), Tur. J. Fish Aqua. Sci., 7: 13-
17.
De Silva SS, Anderson TA (1995). Fish Nutrition in
Aquaculture. Chapman & Hall Aquaculture Series,
London; 319 pp.
Fishman MC (2001). Genomics: Zebrafish--the canonical
vertebrate, 294: 1290-1291.
Goldan O, Popper D, Karplus I (1997). Management of
size variation in juvenile gilthead seabream (Sparus
aurata), I Particle size and frequency of feeding dry
and live food, Aquaculture, 152: 181– 190.
Grayton BD, Beamish FWH (1977). Effects of feeding
frequency on food intake, growth and body
composition of rainbow trout (Salmo gairdneri),
Aquaculture 11: 159-172.
Hevroy EM, Espe M, Waagbo R (2005). Nutrition
utilization in Atlantic salmon (Salmo salar) fed
increased level of fish protein hydrolyses during a
period of fast growth, Aquacult. Nutr., 11: 301-313.
James R, Sampath K (2003). Effect of animal and plant
protein diets on growth and fecundity in ornamental
fish, Beta splendens (Regan), J. Aquacult. Bamid.,
55(1): 39-52.
Jobling M (1983). Effect of feeding frequency on food
intake and growth of Arctic charr, Salvelinus alpinus
(L). J. Fish. Biol., 23: 177- 185.
Kasiri M, Farahi A, Sudagar M (2011). Effects of Feeding
Frequency on Growth Performance and Survival Rate
of Angel Fish, Pterophyllum scalare (Perciformes:
Cichlidae), Vet. Res. Forum, 97- 102.
Kestemont P, Baras E (2001). Environmental factors and
feed intake: mechanisms and interactions, in Houlihan
et al., ed,. Food intake in fish Blackwell Science-COST
Action; 827, Oxford.
Lamason RL, Mohideen MA, Mest JR, Wong AC, Norton
HL (2005). A putative cation exchanger affects
pigmentation in zebrafish and humans, 310: 1782-
1786.
Lee SM, Cho SH, Kim DJ (2000). Effects of feeding
frequency and dietary energy level on growth and
body composition of juvenile flounder Paralichthys
olivaceus (Temminck and Schlegel), Aquacult. Res., 31:
917– 921.
Lim LC, Wong CC (1997). Use of the rotifer, Brachionus
calyciflorus Pallas, in freshwater ornamental fish
larviculture. Hydrobiologia, 358: 269- 273.
Mandal B, Mukherjee A, Banerjee S (2010). Growth and
pigmentation development efficiencies in fantail
guppy, Poecilia reticulata fed with commercially
available feeds. Agricult. Biol. J. N. Am., 1(6): 1264-
1267.