Mirea (Ciortan) C. et. al.

/Scientific Papers: Animal Science and Biotechnologies, 2013, 46 (2)

Results Regarding Growth Performance of Nile tilapia

(Oreochromis niloticus, Linnaeus, 1758) Fed with an Additive
Feed, Vitamin C, in a Recirculating Aquaculture System
Catalina Mirea (Ciortan), Victor Cristea, Rodica Iulia Grecu,
Lorena Dediu, Sarbu Adina

“Dunarea de Jos” University of Galaţi, Aquaculture, Environmental Sciences and Cadastre Department
800008-Galati, Domneasca, 47, Romania

Abstract
The aim of this study was to determine the vitamin C status on growth performance, survival rate and biochimical
parameters from tissue of Nile tilapia (Oreochromis niloticus, Linnaeus, 1758) with average weight of 93±1.0 g were
fed with different dietary vitamin C rations and reared in a recirculating aquaculture system (RAS). Growth
performance , survival rate and biochimical parameters from tissue of Nile tilapia were measured in groups fed diets
supplemented with 100, 750, 2000 mg.kg-1L-ascorbic acid (AA) and almost ascorbate free diet (Total AA>5 mg.kg-
1) as a control. They were stocked in 12 rearing units at 28 oC water temperature for 30 days. Diet of 33% protein
was offered as feed, 3 times daily. Growth measurements of tilapia were recorded on beginning and at the end of the
experiment. Results showed that growth performance for Nile tilapia had not significantly (p>0.05) differences
between the control and the tretments . Survival rate was the same for al the treatments. The feed conversio ratio
(FCR) was not significant (p>0.05) between the control and the treatments. Resuls showed that the feed impoved
with AA had no significant differences between treatments and control regarding the optimum growth performance
and survival rate at Nile tilapia.
Keywords: ascorbic acid, diet, growth performance, Nile tilapia, RAS, treatments, vitamin C

1. Introduction Tilapias showed classical vitamin C deficiency

Vitamins are organic substance essential for
growth, health, reproduction and maintenance in
animals. Since fish cannot synthetize sufficient
quantity for normal development, they have to be
supplied in diet [1].
Vitamin C is an indispensable nutrient for fish, as
they cannot synthesize this nutrient due to the lack
of enzyme L-gulonolactone oxidase; therefore
they depend on exogenous supply through dietary
source [1].

* Corresponding author: Catalina Mirea (Ciortan)
Tel: 0040742069549
Email: catalinamirea@yahoo.com
catalina.ciortan@ugal.ro
signs when fed a vitamin–deficient diet in the
absence of natural foods. The requirement for
normal growth of O.aureus has been reported to
be 50 mg AA/Kg of diet. In juvenile O.niloticus x
O. aureus, a dietary level of 79 mg AA/kg of diet
is needed for maximum growth. A recommended
dietary vitamin C inclusion level for O.niloticus is
420 mg AA/kg of diet [1].
Based on findings on the influence of vitamin C
supplementation on immune response and on
increasing field experience in this important area
of fish nutrition, the following dose is
recommended (1000 mg AA/kg of diet) when the
immune system is challenged (stressing situation,
handling and grading, winter wounds, disease
outbreaks and release of smut into the sea).

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Vitamin C has an important role the intracellular Nile tilapia in agreement with El-Sherif et al.,
and extracellular processing and assembly of (2009) [3].
collagen from its precursors. Collagen is a Figure 1 showed further that DO was in the
principal constituent of the skin, scales, mucus, optimum range for Nile tilapia like El–Sherif et
cartilaginous tissues, and bones in conjunctive al., (2009) [5] showed that the best growth rate
tissue formation, which involves all organs of the DO ranged from 7 to 8.3 mg L-1.
body. Acid ascorbic was also reported to have
many curative effects due to its proprieties, by
many authors. They reported that ascorbic acid
reduced the mortalities, lowered the metal content
of tissue and prevented the inhibition of blood
AST and LDH activity of Tilapia zillii after
exposure of mercury. Abdel-Tawwab et al. [2]
found that a high level of acid ascorbic enhanced
the weight gain, specific growth rate and survival
rate of Nile tilapia exposed to sub lethal dose of
mercury.

2. Materials and methods

The experiment was carried out within the pilot

recirculating system of Aquaculture,
Environmental Science and Cadastre Department,
University “Dunarea de Jos” of Galati.
The experimental activity period lasted for 30
days, from March until April 2012, in twelve
rearing units with a capacity of 45 l/unit
(45×30×39 cm). The system was provided with
mechanical filtration type TETRATEC EX 400.
De-chlorinated water in the rearing units were
aerated by a constant supply air of compressed air
pump type RESUN Quiet LP 100 , and the water
was exchange once a day with 50% of water from
each unit, before morning feeding. Dissolved
oxygen and temperature was daily measured in the
morning with Hannah HI 98186 oximeter, and
NO2-, NO3-, NH3-, were measured with Figure 1 and 2. Mean values of physico-chemical
Spectroquant Nova 400. characteristic of water

Water parameters The second figure showed optimum values for

The recorded values showed suitable
environmental conditions for rearing Nile tilapia
during the experimental period (Figure 1. and
Figure 2.).
These values are in agreement with those of El-
Sherif and al., (2009) [3] who noted that the
optimum range for growth and food conversion
was 21-28oC. The values of pH during the
experiment was 6.80 the minimum and 7.95 the
maximum this values are in the optimum range of
nitrite, nitrate and ammonium for growth
performance and food conversion.
Experimental fish. Nile tilapia (Oreochromis
niloticus) used in this experiment were 93±0.66 g
mean average weight and 17±0.8cm of length.
Fish were homogenous in size, body weights and
healthy. They were fed on the same diet like in the
experiment for 2 weeks before the study to adapt
them to the experiment conditions. Through the

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adaptation period the week and the dead fish were
eliminated daily.
Experimental design. The fish was stocked into
12 rearing units at 100, 750, 2000 mg.kg-1L-
ascorbic acid (AA) and almost ascorbate free diet
(Total AA>5 mg.kg-1) as a control. Automatic
heaters with thermostat were used to obtain the
temperatures. The number of tilapia stocked in the
rearing units was in a rate of 10 fish per unit for
30 days with three replicates for each treatment.
Commercial diet was used in the experiment; the
pellets size was 2 mm, which had 33% crude
protein.
The daily feeding tare was 2% of the total
stocking biomass according to the feeding chart;
the fish were fed three times daily.
The feed was offered spreading by hand in the
unit. The AA used was provided from Janssen
Chimica Company (Geel, Belgium) and had
99.9% purity. Working protocol of incorporation
Vitamin C in feed following these steps:
 Dissolution of Vitamin C in gelatin solution
2%;
 Uniformly spraying the final solution on
thesurface of feed grains by continuous
agitation;
 Drying the pellets incorporated with probiotic
at To=25oC for 2 h.
The determination of vitamin C from tissue at the
end of the experiment was established with a
chemical method (Riemschneider et al., 1976) as a
simplified method for the simultaneous
determination of the total vitamin C, DNPH
method [4].
Growth parameters. The following parameters
were used to evaluate tilapia growth performance:
 Weight gain (W)=Final Weight (Wt)–Initial
Weight (W0) (g);
 Individual Weight Gain (IWG) (g/ex)=(Final
Weight (Wt)–Initial Weight (Wo))/Total Fish
Exemplars;
 Food Conversion Ratio (FCR)=Total feed
(F)/Total Weight Gain (W) (g/g);
 Specific Growth Rate (SGR)=100 × (ln Wt–ln
Wo)/t (%BW/day);
 Protein Efficiency Ratio (PER)=Total Weight
Gain (W)/Amount of protein fed (g) ;
Processing the statistical data
Statistical analysis was performed using t test
(p=0.05) and ANOVA (Microsoft Corporation).
240
The coefficient of variation (CV) was calculated
as the ratio of the standard deviation to the mean
in order to have a measure of dispersion.
3. Results and discussion
Technological indicators of Nile tilapia growth
obtained at the end of the experimental period are
summarized in Table 1 and graphically presented
in Figure 3. The following is a critical analysis of
the most significant technological indicators,
namely, specific growth rate (SGR) and feed
conversion ratio (FCR).
Figure 3. FCR, SGR and PER after
the treatment with vitamin C
Growth performance parameters
At the end of the experimental period, the first
three variants (V1, V2, V3) for the biomass of the
culture we can see that we have the same biomass
for all the treatments applied.
It is known that between efficiency of feed
utilization factor expressed by the feed .For this
experiment we observe (Figure 3) similar values
of FCR's for versions V1, V2, V3 (0.56, 0.57,
0.57), trend that is observed by SGR too or a
better characterization of fish biomass was
calculated coefficient of variation (Table 1).
Increased growth biomass was proportional to the
initial biomass, but no significant difference in
statistical terms. Subject to the application of the
same intensity of feeding in all cases, was
followed, in the experiments performed to identify
a possible correlation between another important
indicator conversion factor protein (PER) and feed
conversion factor (Figure 3).
From the graphical representation of these two
parameters can be seen primarily as feed
conversion factor is maximum in variant 3 (V3),
 Mirea (Ciortan) C. et. al./Scientific Papers: Animal Science and Biotechnologies, 2013, 46 (2)

as well as the other growth parameters. It is worth mentioning also that the variation in the
conversion factor of the food on the four factor protein.
parameters was symmetrical with the conversion

Table 1. Technological indicators for Nile tilapia at the end of the experiment
Control V1 V2 V3
(100 mg AA/Kg (725 mg AA/Kg (2000 mg AA/Kg
comercial feed) comercial feed) comercial feed)
B1(Avarage) B2(Avarage) B3(Avarage) B4(Avarage)
Initial numbers of fish 10.00 10.00 10.00 10.00
Final numbers of fish 10.00 10.00 10.00 10.00
Survival (%) 100.00 100.00 100.00 100.00
Initial biomass (g) 932.67 928.00 923.33 926.33
Final biomass (g) 2110.67 2099.33 2093.67 2093.00
Biomass gain (g) 1178.00 1171.33 1170.33 1166.67
Mean individual weight (g/ex) 93.27 92.80 92.33 92.63
Mean final fish weight (g/ex) 211.07 209.93 209.37 209.30
Specific growth rate SGR (%/day) 2.72 2.72 2.73 2.72
Feed conversion ratio FCR (g/g) 0.57 0.57 0.57 0.57
Protein efficiency ratio PER ((g/g) 5.30 5.29 5.30 5.27
*the values from Table 1 were statistically analysed (p=0.05), and we can conclude that are not significant
differences between the groups, the technological indicators indicate that SGR, FCR and PER was almost the same
in the entire group.

From the analysis presented in a graphical form,

growth equations for each of the experimental
variants (Figure 4).

Figure 4. Weight–Lenght regression

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Two main conclusions can be drawn. The first
refers to the fact that there is no difference
between the relatively high allometric factors of
each variant the nearest being optimal was V3
(2000 mg AA/kg feed). The second conclusion
drawn from equation R2 sensitive growth
parameter equal terms in all breeding which
expresses uniformity coefficient condition in all
experimental variants.
Estimation of vitamin C from tissue
After determination of the λ max of the colored
complex (521 nm) using a UV- spectrophotometer
the absorbance of the all standards (converted to
colored complex) were taken to construct a
calibration curve. The calibration curve was
constructed by plotting the concentration versus
the corresponding absorbance.
Figure 5. Calibration curve of standard vitamin C
4. Conclusions
Doing the critical analysis of all technological
indicators in this experiment, it can be concluded
that the level of vitamin C in feed is not directly
proportional to the growth performance in a
recirculating system.
We also can conclude that the individual growth
in Variant 2 (V2) was not uniform like in the
others variant how we can see in Figure 4, the
regression show that the growth is nonlinear for
this group.
Due to its vitamin C on the immune system is the
main factor in preventing nutrition, based on the
presented in this paper we assert that statistical
calculations are not distinguished by any dose of
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The ascorbic acid concentration vary with
conditions such as temperature and the storage
period on peservation.
It is observed that vitamin C content slowly
decreases with temperature and storage period.
The result showed that vitamin C retained in fish
tissue is deacresing with increasing the value of
the dose included in the feed (Figure 6).
To optimize the stability of vitamin C during the
feed production and storage, it is recommended to
use a form of it phosphors. The recommended
dose of vitamin C for trout, salmon and catfish is
1000 mg AA/kg feed (Viviane Verlhac and
Jacques Gabaudan et al., 2003) [6], which led us
to the choice of larger amounts or lower than this
optimum in this experiment.
Figure 6. Concentration of vitamin C from
samples
vitamin C that we can add the feed to enhance
feeding or to have important technological
performance, however we can say that the
addition of AA on fish had a beneficial effect on
mortality is not registered throughout the
experiment and also any illness.
In the literature we can say that we recommend a
diet supplemented with vitamin C occurred
whenever the disease in the growth of biomass
crops.
Because vitamin C cannot be synthesized by the
fish we see that there are minimal amounts of this
vitamin in any commercial feed.
Finally recommend the use of vitamin C in feed
after each period of technological stress and low
immunity after a starvation period or a treatment.
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Acknowledgements
The work of Catalina Mirea (Ciortan), was supported
by Project SOP HRD – TOP ACADEMIC 76822/2010,
funded by the European Union and the Romanian
Government. The authors particularly thank the project
management team for the financial support.
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