Scientific Papers. Series D. Animal Science. Vol. LXIV, No.

1, 2021
ISSN 2285-5750; ISSN CD-ROM 2285-5769; ISSN Online 2393-2260; ISSN-L 2285-5750

THE INFLUENCE OF STOCKING DENSITIES ON GROWTH

PERFORMANCE OF COMMON CARP (CYPRINUS CARPIO, LINNE 1758)
REARED IN A RECIRCULATING AQUACULTURE SYSTEM

Anca Nicoleta CORDELI (SĂVESCU), Lucian OPREA,

Mirela CREȚU, Mihaela MOCANU

“Dunărea de Jos” University of Galaţi, 47 Domnească Street, 800008, Galați, Romania

Corresponding authors emails: [Link]@[Link]; [Link]@[Link]

Abstract

To determine the influence of stocking density on the common carp growth performance and body composition, four
variants: 0.9 kg m-3 (V1), 1.8 kg m-3 (V2), 2.6 kg m-3 (V3), and 3.5 kg m-3 (V4) was carried out during 31 days. In this
context, 2100 common carp fry, with an initial weight of 1.8 g fish-1, were maintained in twelve rearing units of a
recirculating aquaculture system. Fish were fed three times per day with extruded feed containing 50% crude protein
and 14% fat. At the end of the experiment, the fish's growth performance was assessed. Better results were obtained for
the V1 variant. Regarding the biochemical composition of fish meat, significant differences (p˂0.05) were recorded in
the water content, while the ash lipids and protein content showed no significant differences (p˃0.05) between the
experimental variants. Therefore, it can be concluded that a stocking density of 0.9 kg m-3 is optimal for rearing
juveniles of common carp without compromising the specific growth rate (SGR), survival, and biochemical composition
of fish meat.

Key words: biochemical composition, FCR, SGR, stocking density.

INTRODUCTION these systems depends on the density at which

the fish are stocked (North et al., 2006).
According to FAO, aquaculture is contributing Therefore, determining the optimum stocking
more and more to world production of aquatic density is the most important criterion for
food, given that, in the case of the wildest fish designing an intensive aquaculture system
stocks, the limits of sustainable exploitation are (Summerfelt & Vinci, 2008).
at present almost touched or even outdated. In Higher stocking densities can affect digestion
2018, 46% of the total production and 52% and food absorption (Abdel-Tawwab et al.,
from fish for human consumption were assured 2014), reduce fish growth performance,
from aquaculture (FAO, 2020). survival, size variation, health, and fish
In Romania, aquaculture is predominantly mortality (Ruane et al., 2002; Pouey et al.,
freshwater, with the availability of inland 2011). Also, higher stocking densities lead to
waters providing the right conditions for fish deterioration of water quality because of the
farming. According to FAO Fishstat, in 2018, metabolic excretion of fish (Çağiltay et al.,
the aquaculture sector from Romania produced 2017), increase stress (Aksungur et al., 2007),
12298 tons. an aspect which can have consequences in the
The most important cultured fish species from aggressive behavior of fish. According to a
our country is common carp (Eurostat, 2017). study by Firas et al. (2020), at higher stocking
Usually, it is raised extensively or semi- densities, fish spent more time feeding and
intensive in polyculture with Asian cyprinids. swimming and less time resting, an aspect that
These growing technologies are based on the can negatively affect fish growth. On the other
natural productivity of the ponds with some hand, lower stocking densities may reduce the
additional feed based on local cereals. overall production (Apu et al., 2012), causing
Lately, there is also an increasing interest economic losses.
regarding the rearing of carp in intensive Various studies have been carried out about the
conditions, mainly in recirculating aquaculture effects of stocking density on fish growth
systems (RAS). Generally, the profitability of performance for different species such as
509
rainbow trout (Sirakov & Ivancheva, 2008; Simultaneously, the concentration of nitrogen
Mocanu et al., 2011; McKenzie et al., 2012), compounds was measured twice per week with
Atlantic salmon (Wang et al., 2019), African the help of the Spectroquant Nova 400
catfish (Van de Nieuwegiessen, 2009) and so photometer with Merck kits.
on. Positive or negative effects on growth
performance have been reported from these Table 1. Ingredients of the experimental diet
studies, and the pattern of this relationship
Composition Quantities
appears to be species-specific. That is why, to
Crude protein 50%
maximize the production and profitability of a Fat 14%
RAS system, it is important to determine the Crude cellulose 2%
optimum stocking density suitable for each Digestible energy 4100 kcal kg-1
species and each growing stage. Lysine 2.5%
Although carp is a fish raised worldwide, the Phosphor 1%
information about the optimal stocking density Copper 6 mg
Vitamin A 20000 UI kg-1
practiced in the RAS is limited or non-existent. Vitamin D3 2000 UI kg-1
Therefore, this study aimed to investigate the Vitamin E 200 mg kg-1
effects of stocking densities on the growth Vitamin C 200 mg kg-1
performance, survival, and biochemical Ingredients: fish meal, poultry meal, corn gluten,
composition of common carp, with an initial wheat gluten, wheat flour, animal fat, feed yeast,
weight of 1.8 g, reared in a recirculating hemoglobin, vitamins, minerals.
aquaculture system.
Fish growth performance. At the end of the
MATERIALS AND METHODS experiment, the following technological
efficiency indicators were calculated: growth
Experimental design. The present study was rate, food conversion ratio, specific growth
conducted for 31 days in a recirculating rate, and the protein efficiency ratio using the
aquaculture system (RAS) at the Faculty of following equations:
Food Science and Engineering, University ✓ Weight Gain (W) = Final Weight (Wt) –
Dunărea de Jos, Galați, România. The RAS Initial Weight (W0) (g);
system is provided with twelve rearing units, ✓ Food Conversion Ratio (FCR) = Total feed
with a volume of 0,132 m3 each. The (F)/Total weight gain (W) (gg-1);
recirculating system was described in detail in ✓ Specific Growth Rate (SGR) (%Body
the paper of Mocanu et al. (2011). weight day-1)) = [(LnWt–LnW0)/t] × 100.
A total of ț2100 fishes with an initial weight of Somatic measurements were made at the end of
1.8 g fish-1 were stocked in the rearing units of the trial at 50 fish/experimental variant. Total
the RAS system to create four experimental length (TL) and body weight (BW) for each
variants: V1-70 fish, and the initial stocking variant were used to determine the relationship
density of 0.9 kg m-3, V2-140 fish, and the W=a×Lb, where “a” is the intercept (the initial
initial stocking density of 1.8 kg m-3, V3-210 growth coefficient), and “b” is the allometric
fish and the initial stocking density of 2.6 kg coefficient (Ricker, 1975). The coefficient of
m-3, and V4-280 fish and the initial stocking variation (CV, %) was calculated as the ratio of
density of 3.5 kg m-3. The experiment was the standard deviation to the mean of weight to
conducted in triplicate. have a measure of fish dispersion.
Fish were fed three times per day with a diet Proximate analysis of fish. To determine the
containing 50% crude protein and 14% fat at a biochemical composition of fish, samples were
feeding level of 5% BW day-1 (Table 1). taken both in the initial and final stages of the
During the experiment, fish were kept under a experiment. The proximate composition of fish
natural photoperiod of approximately 12/12 h was analysed using the AOAC (2000) method.
light/dark cycle. The chemical composition of meat crude
Water quality parameters such as dissolved protein was analysed according to the Dumas
oxygen, temperature, and pH were recorded method (N×6.25), and crude lipids were
daily with the help of Hannah 98194. determined by the Soxhlet method, using
petroleum ether as a solvent.
510
Dry matter was determined by drying the Kolmogorov-Smirnov tests determined the
samples at 105 ± 2ºC using Jeio Tech normality of the data used for analysis. One-
Convection Oven, and ash was evaluated by way ANOVA and Duncan’s multiple range
calcification at temperatures of 550 ± 20°C in a tests were used to compare the differences
Nabertherm furnace. between the experimental groups. Significance
The main indicators used for the evaluation of was determined at α = 0.05.
biochemical fish compositions were as follows:
✓ Protein efficiency ratio (PER): PER = RESULTS AND DISCUSSIONS
(Bf-Bi)/(F×PB), where: F = quantity of
administrated fed (kg), PB = amount of fed Water quality. It is well known that water
protein (%); quality has a significant impact on the fish's
✓ Protein utilization efficiency (PUE): PUE = biology and physiology, affecting the health,
100×(W×Pf–Wi×Pi)/(F×Pb) (%), where: Pf welfare, and productivity of a fish culture
- muscle tissue protein at the end of the system. Generally, at higher stocking densities,
experimental period (%); Pi - muscle tissue lower growth performance is also determined
protein at the initial stage of the by an increased production waste production
experimental period (%); Wf - final biomass rate.
(kg); Wi - initial biomass (kg); F - total feed Water chemical parameters during the
quantity consumed (kg); Pb - administrated experimental period are presented in Table 2.
feed protein concentration (%). In our recirculating system, all the water
Data analysis. Data were analysed using SPSS parameters were reasonably constant during the
21 for Windows. Data regarding fish growth experimental period and were not affected by
performance and the biochemical composition stocking density (ANOVA, p˃0.05).
were expressed by average and standard
deviation (Average ± SD).

Table 2. Synthetic table with the average values (± SD) of the main physicochemical parameters of water
Parameters V1 V2 V3 V4
Temperature °C 21.2±0.18 21.6±0.11 21.4±0.12 21.6±014
Dissolved oxygen (mg L-1) 7.71±0.12 7.23±0.09 7.10±0.12 7.05±0.08
pH (pH units) 7.62±0.11 7.32±0.09 7.41±0.11 7.29±0.10
N-NO3- (mg L-1) 21.13±0.20 19.1±0.7 18.5±0.36 17.9±0.65
N-NO2- (mg L-1) 0.03±0.02 0.04±0.01 0.05±0.02 0.05±0.01
N-NH4+ (mg L-1) 0.17±0.02 0.22±0.05 0.19±0.03 0.19±0.04
P2O5 (mg L-1) 5.90±0.19 5.53±0.35 5.2±0.43 5.76±0.32
Note: Data are presented as triplicate mean ± SD.

Water temperature was around 21.2 ± 0.18 °С were cleaned daily, and only 10% of fresh
in V1, and 21.6 ± 0.14 °С in V4, dissolved water was added to the RAS system. Also, a
oxygen content varied between 7.05 ± 0.08 mg significant role for the maintenance of optimum
L-1 in V4 and 7.71 ± 0.12 mg L-1 in V1. Water water chemical parameters in RAS during the
pH in the recirculation system was kept experimental period has the mechanical and
constant, and the lowest values were recorded biological filters which conditioned the water
in the V4 variant (7.29 ± 0.10 pH units). Also, properly.
the nitrogen compounds were in the optimum Fish growth performance. The mean final
interval for the cultivated fish species. The weight, mean weight gain, percentage survival
maintenance of these optimum water concen- rates, FCR, and SGR of fish in all the
trations was possible by the benefit of treatments at the final of the experiment are
optimized technical and technological presented in Table 3.
parameters of the RAS. The fish rearing units

511
 Table 3. Technological performance indicators obtained at the end of the experimental period
Growth performance V1 V2 V3 V4
Initial biomass (g) 126 252 378 504
Initial biomass (kg m-3) 0.9 1.80 2.60 3.50
The initial number of fish 70 140 210 280
Initial weight (g fish-1) 1.80±0.00 1.80±0.00 1.80±0.00 1.80±0.00
Final biomass (g) 313.67±6.66 521±12.77 692±20.52 879.67±28.50
Final biomass (kg m-3) 2.20±0.05 3.65±0.09 4.84±0.14 6.16±0.20
The final number of fish 68.00±1.0 122±3 171.67±6.51 221.67±6.03
Final weight (g fish-1) 4.61±0.03* 4.27±0.08** 4.03±0.15*** 3.97±0.02***
Weight gain (g) 187.67±6.66* 269.00±12.77** 314±20.52*** 375.67±28.50****
Weight gain (kg m-3) 1.31±0.05* 1.88±0.09** 2.20±0.14*** 2.63±0.20****
Individual weight gain (g) 2.81±0.03* 2.47±0.08** 2.23±0.15*** 2.17±0.02***
Survival rate (%) 97.14±1.43* 87.14±2.14** 81.75±3.1*** 79.17±2.15***
SGR (% day-1) 2.94±0.07* 2.34±0.08** 1.95±0.10*** 1.80±0.10***
FCR(g g-1) 1.84±0.06* 2.57±0.12** 3.31±0.22*** 3.69±0.28****
Note: Data are presented as triplicate mean ± SD.

Despite the fact that all the water parameters 10

were adequately maintained in all treatment
8
Individual weight (g)
groups, in the present study, stocking density
affects the growth performance of common 6 y = 0.06x2.46
carp, being observed a negative correlation 4 R² = 0.82
between stocking density and growth
2
performance.
The data obtained for each experimental variant 0
0 2 4 6 8 10
regarding the final fish weight showed no
Total length (cm)
deviations from the normal distribution (p>0.05
with Kolmogorov–Smirnov test) that permitted Figure 1. Length-weight regression at the end of the
us to apply the parametric tests further. One- experiment for the V1 variant (n = 50 fish)
way ANOVA used at the end of the experiment
10
showed significant differences between the
final weight of fish (ANOVA, p˂0.05). So, the
Individual weight (g)

8
y = 0.04x2.59
mean final weight of fish at the end of the 31- 6 R² = 0.80
day experimental period was as followed: V1-
4
4.61±0.03 g, V2-4.27±0.08 g, V3-4.03±0.15 g,
and V4-3.97±0.02 g. The post hoc Duncan 2
analysis showed that the final weight of fish 0
from V1 and V2 was significantly higher than 0 2 4 6 8 10
Total length (cm)
those of fish from V3 and V4.
At the end of the experiment, total length (TL)- Figure 2. Length-weight regression at the end of the
weight (W) regressions were plotted (Figures experiment for the V2 variant (n = 50 fish)
1-4) to obtain more information about the
growth patterns of the fish. 10
The slope (b) values obtained for all the y = 0.03x2.74
Individual weight (g)

8
experimental variants showed a negative R² = 0.87
6
allometric growth indicating that the fish length
was higher than the body mass. However, in 4
this study, the length-weight relationship was 2
found to be highly correlated, and all values of
0
the coefficient of determination were greater
0 2 4 6 8
than 0.80. Total length (cm)

Figure 3. Length-weight regression at the end of the

experiment for the V3 variant (n = 50 fish)

512
 significantly higher in V1 (2.94±0.07% day-1).
10
The post hoc analysis showed that the SGR
8
y = 0.04x2.58 values from V1 were higher than V2
Individual weight (g)

6 R² = 0.94 (2.34±0.08% day-1), while in V3 (1.95±0.10%

4
day-1) and V4 (1.80±0.10% day-1), the values
are similar (p˃0.05).
2 Regarding FCR, Duncan's multiple range test
0 divided the obtained values into four distinct
0 2 4 6 8 10 groups, the best values being obtained in V1.
Total length (cm)
The FCR ranged from 1.84±0.06 in V1 to
Figure 4. Length-weight regression at the end of the 3.69±0.28 in V4 and increased with an increase
experiment for the V4 variant (n = 50 fish) in fish stocking density. Therefore, the higher
The coefficient of variation (CV) showed FCR values obtained at the highest stocking
higher variability in the V4 variant densities indicate low food utilization
(36.35±3.88%), followed by the V2 efficiency.
(32.66±7.06%) and V3 (32.53±3.10%). The In the present study, a negative correlation was
lowest CV was obtained in V1 observed between the stocking density and fish
(CV=24.47±3.11%). growth performance. The effect of stocking
Results showed that increasing stocking density density on growth is in line with the results
results in higher variation in individual growth. obtained for other cultured fish species.
An increase of the CV over time indicates HtayHtay et al. (2019) conducted a study over
inter-individual competition within the fish five months stocking carp as follows (with an
group (Azaza et al., 2013). Obtaining a higher individual weight of 0.5-1.6 g, and standard
variability at higher stocking density is length 2.2-4.9 cm): 5 fish per tank, ten fish per
undesirable in aquaculture, preferable to reduce tank, and 15 fish per tank (water volume 40
fish size variations and obtain homogeneous L/tank). After five months, the best results for
fish size, which facilitates feeding, harvesting, fish survival and growth performance were
marketing, and processing (Azaza et al., 2010; observed in the variant with the lowest stocking
Azaza et al., 2013). density (5 fish/tank). Also, Marandi et al.
The individual weight gain of fish was (2018), reported after 45 days of growing,
significantly affected by stocking density better values of FCR and SGR for common
(ANOVA, p<0.05). Duncan's multiple range carp (initial weight of 1.41±0.5 g fish-1) at
tests showed three distinct groups: the lower stocking densities (20 fish/tank, or 0.70 g
individual weight gain of fish from V1 was L-1).
significantly different from those of V2. In The proximate composition of common carp
contrast, the individual weight gain of fish from reared at different stocking densities is
V3and V4 was similar. presented in Table 4. Generally, fish's
Survival is a crucial indicator of fish health biochemical composition is influenced by many
status (Rey et al., 2019). In our experiment, it factors that depend especially on species, size,
can be observed that the survival rate was age, environmental conditions, and feeding
directly influenced by the stocking density, and (Cho, 2001).
significant differences (ANOVA, p˂0.05) were The results showed significant differences
obtained. The post hoc analysis showed that the (ANOVA, p˂0.05) in fish water content the
survival rate of fish from the V1 (97.14±1.43%) percentage between the four stocking densities.
group was significantly higher than that from So, the water content from V1 and V2 was
V2 (87.14±2.14%), while no significant significantly different from the water content
differences (p˃0.05) were recorded between the from V3 and V4 variants. A significant increase
V3 (81.75±3.1%) and V4 (79.17±2.15%) in water content was observed in V1, V3, and
groups. V4 variants compared with the initial moment.
Also, in SGR and FCR, the best values were Regarding the protein, lipids, and ash content,
obtained in the lowest stocking density (V1). no significant differences (ANOVA, p˃0.05)
The average specific growth rate was were recorded between the four stocking

513
densities, but significant differences were significant increase in protein and lipids'
recorded compared to the initial moment content and a significant decrease in ash
(ANOVA, p˂0.05). Thus, there was a content.

Table 4. The proximate composition of common carp meat reared at different stocking densities
Experimental variants
Parameters
Initial V1 V2 V3 V4
Water (%) 76.11±0.18 76.48±0.07ac 76.33±0.22 75.20±0.22 75.32±0.08
Protein (%) 12.29±0.18 13.15±0.02bc 13.26±0.02 13.15±0.16 13.53±0.47
Water/protein 6.19±0.03 5.81±0.05bc 5.75±0.02 5.71±0.05 5.57±0.20
Lipids (%) 7.78±0.07 8.80±0.08bc 8.71±0.2 8.81±0.21 8.63±0.21
Ash (%) 2.19±0.10 1.42±0.02bc 1.39±0.07 1.46±0.03 1.44±0.03
Note: Data are presented as triplicate mean ± SD;
a-significant differences between the experimental variants (p<0.05); b-insignificant differences between the experimental variants (p>0.05).
c-significant differences from the initial moment (p<0.05); d-insignificant differences from the initial moment (p<0.05)

The water to protein ratio is a suitable values from V1 were significantly different
instrument to detect excessive water, being (p˂0.05) from those obtained in V2, while in V3
more precise and reliable than the water content and V4, no significant differences were
itself (Manthey-Karl et al., 2012). If the value obtained in the PER values.
of the Water/Protein ratio is lower, the Also, significant differences (ANOVA, p˂0.05)
nutritional value is higher. The applied stocking were obtained between the values of protein
densities on common carp have no significant utilization efficiency. The evolution of PUE
differences (ANOVA, p˃0.05) in the water to emphasizes a better protein valorisation
protein ratio. Compared with the initial inversely proportional to the increase of the
moment, the nutritional value of fish meat stocking density. Duncan's multiple range tests
reflected by the water/protein ratio was revealed four distinct groups belonging to each
significantly better (ANOVA, p˂0.05). At the tested stocking density.
beginning of the experiment, the water/ protein Our values obtained by us regarding carp
ratio was 6.19±0.03, and at the end of the carcass's biochemical composition are similar
experiment, this ratio decreased for all groups, to those obtained from other authors. In a study
reaching 5.81±0.05 in the V1, 5.75±0.02 in V2, conducted by Khushwinderjit et al. (2018), the
5.71±0.05 in V3, and 5.57±0.20 in V4. biochemical composition of flesh of the
To have more information regarding the fish common carp fingerlings (with the weight
protein gain, we calculate the protein efficiency between 5.41-5.49 g, and length between 6.61-
ratio (PER) and the productive protein value 6.74 cm) fed with diets replacing protein of
(PUE) (Figure 5). plant origin with animal protein in the form of
fish silage at different levels, was as follows:
water content ranged between 78.20-81.43%,
crude protein 13.90- 16.50%, fat 1.60-2.50%,
and ash between 1.06-1.60%.

CONCLUSIONS

The effects of stocking density were evident in

the growth of common carp in weight gain and
the final weight of fish. The best stocking
density concerning growth performance and
Figure 5. The protein efficiency ratio (PER) and protein feed conversion efficiency was at 70 fish per
utilization efficiency (PUE) rearing unit, with the initial stocking density of
0.9 kg m-3. However, the final results regarding
ANOVA test revealed significant differences the fish-stocked weight were still low (0.9 kg
(p˂0.05) in PER and PUE values. Duncan's m-3). This study, therefore, recommends further
multiple range tests showed that the PER
514
research with lower stocking densities, which FAO, Food and Agriculture Organization of the United
would result in higher final productions. Nations. (2018). The State of World Fisheries and
Aquaculture 2018 - Meeting the sustainable
development goals. Rome, 210 p.
ACKNOWLEDGMENTS Firas, A.M., Nizar, J.H., Nareen, S.A.A., & Salam, R.H.
(2020). Effect of Different Stocking Densities on
The principal author of the article thanks to the Behavior of Common Carp (Cyprinus carpio) in
“Dunărea de Jos” University of Galați, which Duhok Province, Kurdistan Region, Iraq.
through the University Degree Program and the International Journal of Scientific Research in
Biological Sciences, 7(4), 1‒6.
doctoral studies contract has supported the Horwitz, W., & AOAC-Association of Official
achievement. Also, the authors are grateful for Analytical Chemist (2000). Official Methods of
the technical support offered by MoRAS AOAC International. 17th edition, Volume 2, Chapter
through the Grant POSCCE ID1815, cod SMIS 49, 1-64. Gaithersburg, USA: AOAC International
48745 ([Link]). Publishing House.
HtayHtay, W., Sandar, M., & Nyunt, L. (2019). Effect of
stocking density on the growth rate of common carp
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