Aquaculture 479 (2017) 564–570

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Aquaculture
journal homepage: www.elsevier.com/locate/aquaculture

Effect of the transfer at different moments of juvenile Nile tilapia MARK

(Oreochromis niloticus) to the biofloc system in formation
Gabriel Francisco de Oliveira Alvesa, Arthur Francisco Araújo Fernandesb,
Érika Ramos de Alvarengaa, Eduardo Maldonado Turraa, Alexandre Benvindo de Sousaa,
Edgar de Alencar Teixeiraa,⁎
a
Departamento de Zootecnia da Escola de Veterinária da Universidade Federal de Minas Gerais, Av. Antônio Carlos, n0. 6627, Caixa Postal 567, Campus da UFMG, CEP
30123-970 Belo Horizonte, MG, Brazil
b
University of Wisconsin – Madison, 470 Animal Science Building 1675, Observatory Dr., Madison, WI 53706, USA

A R T I C L E I N F O A B S T R A C T

Keywords: Among the production systems used in aquaculture, biofloc systems are gaining prominence due to their need for
Intensive systems little or no water exchange and their high stocking densities. Considering the potential of these systems for the
Biofloc technology production of tilapia, the aim of this study was to evaluate the effect of different moments of the transfer of Nile
Body composition tilapia fingerlings to the biofloc system. For this study, 1400 Nile tilapia fingerlings weighing 1.40 ± 0.04 g
Nile tilapia performance
were distributed into 20 rectangular 150 L tanks in a completely randomized design with five treatments
(Control – clear water, T1 – transition on day 1, T2 – transition on day 15, T3 – transition on day 30, and T4 –
transition on day 45) and four replications. The study lasted 76 days, and the animals remained under the same
control conditions until the time of transition into the biofloc system. The data obtained were subjected to
analysis of variance, and the means were compared with the Student-Newman-Keuls test (p < 0.05). The
average data of final weight (46.16 g), daily weight gain (0.59 g) and feed conversion rate (1.08) did not differ
(p > 0.05) between treatments. The final density was higher in the control treatment (17.44 kg m− 3). The
densities in the T1, T2, T3 and T4 treatments were 10.20, 12.47, 12.68 and 11.49 kg m− 3, respectively. Survival
did not differ according to the Kruskal-Wallis test (p < 0.05); however, the body composition was affected by
the biofloc system, with a greater deposition of lipids in T1. The water quality parameters varied over time, with
N-NO2− peaks in the treatments transitioned to the biofloc system regardless of the time of transition. The
results suggest that Nile tilapia fingerlings should be transitioned to the biofloc system at a weight of 1.4 g and
that they do not require prior cultivation in another type of system.
Statement of relevance: Considering the potential of biofloc systems for the production of tilapia, this study
evaluated the effect of different transition times of juvenile Nile tilapia (O. niloticus) to this systems showing that
this species did not require prior cultivation in another type of system after masculinization.

1. Introduction
The growth and expansion of aquaculture has led to increasing
concern on the impact of these systems on the environment. Because
traditional cultivation systems use generally large volume of water to
maintain quality, untreated effluents might be released into adjacent
environs. Such systems could be potential threats for eutrophication
causing pollution and even spread the diseases (Boyd, 2003; Wetzel,
2001). Recently to reduce environmental impact of aquaculture system
biofloc technology (BFT) has been promising (Avnimelech, 2012).
These cultivation methods are to known for high fish stocking densities
with little to no water exchange (Avnimelech, 1999) instead equipped
for aeration and water agitation to produce aerobic and heterotrophic
biota aggregates of microbial flocs (i.e., bacteria, microalgae, protozoa,
rotifers, feces, food remains and dead animals) (Avnimelech, 2007; De
Schryver et al., 2008).
It has been shown that the microbial community present in the
biofloc contributes in two main ways: maintenance of water quality and
nutrition. In the first case, the community aids in the direct assimilation
of inorganic nitrogen to produce microbial protein (Avnimelech, 1999;
Ebeling et al., 2006). This protein promotes nutritional gain, serves as
part of the diet of the cultured organisms, and may contribute sub-
stantially on improvement of feed conversion, which reduces the pro-
duction costs (Emerenciano et al., 2013). For these reasons, this system

⁎
Corresponding author.
E-mail address: edgarvet@ufmg.br (E. de Alencar Teixeira).

http://dx.doi.org/10.1016/j.aquaculture.2017.06.029
Received 23 August 2016; Received in revised form 17 June 2017; Accepted 19 June 2017
Available online 20 June 2017
0044-8486/ © 2017 Elsevier B.V. All rights reserved.
G.F. de Oliveira Alves et al.
is assumed to be an alternative allowing sustainable development of
aquaculture while simultaneously offering responses to environmental,
social and economic concerns (Crab et al., 2012; De Schryver et al.,
2008).
Studies have demonstrated the feasibility of tilapia cultivation in
biofloc systems because they reduce water, space and food require-
ments, which can become a limiting factor in intensive aquaculture
systems (Avnimelech, 1999; Ekasari et al., 2015; Luo et al., 2014;
Milstein et al., 2001; Avnimelech, 2007; Azim and Little, 2008; Choo
and Caipang, 2015). However, biofloc systems may be more advanta-
geous to tilapia mainly during their juvenile stages. However, little
information is available on growth response of early tilapia on biofloc.
Since the vast majority of commercial production units for Nile tilapia
fingerlings offer masculinized animals weighing from 0.5 to 2.0 g, it
may be more advantageous to receive the fingerlings and direct in-
troduction into the biofloc system to greater control of production and
use of higher stocking densities compared to nurseries. Thus, when
receiving the fish and growing in other systems for the production of
juveniles, such as nurseries that adopt low storage densities and allow a
limited control of the growing conditions, the results can be adversely
affected. With the reduction of the maintenance time in other cropping
systems and a shorter time for the transfer of the fingerlings to the
biofloc system, higher storage densities can be used, together with the
adequate control of water quality parameters and reduction of the
consumption of water and land (Avnimelech, 2012). Thus, this study
evaluated the effect of different transfer times of Nile tilapia juveniles
(Oreochromis niloticus) from a system where water changes were made
to maintain the culture conditions for the biofloc system without water
exchange, evaluating the aspects water quality, animal performance,
survival and body composition of the animals produced.
2. Materials and methods
2.1. Location
The experiment was conducted in a greenhouse belonging to the
Aquaculture Laboratory of the Veterinary School of the Federal
University of Minas Gerais (Universidade Federal de Minas Gerais -
UFMG), Brazil. All procedures were previously approved by the Ethics
Committee on Animal Use of UFMG under protocol number 98/2013.
2.2. Fish and experimental conditions
For the experiment, 1400 masculinized Nile tilapia (O. niloticus)
fingerlings were used. The animals were distributed into 20 rectangular
polypropylene tanks with volumes of 150 L each at a density of 70 fish
per tank; the fish altogether had an initial mean biomass of
98.10 ± 3.08 g, with a mean initial weight of 1.40 ± 0.04 g.
The study lasted 76 days and followed a completely randomized
design, with five treatments and four replications for a total of 20 ex-
perimental units. Each tank had a 300 W heating thermostat fixed at
28 °C water temperature in the tanks to reduce temperature variations
throughout the experimental period. The treatments evaluated were
four transitional moments of the fish of a system where water changes
were made to maintain the culture conditions for the heterotrophic
medium, initiating the formation of biofloc, besides to the treatment
that remained during all study with water changes (designated as
control). In the control treatment, the animals were maintained in a
clear water system with daily water changes to maintain quality. The
other treatments were as follows: T1, the animals were subjected to the
formation of microbial flocs (bioflocs) beginning on the first day of the
experiment (mean initial weight at the initiation of biofloc formation of
1.4 ± 0.04 g; T2, the animals remained under control conditions until
the 15th day, at which time biofloc formation was initiated (mean
weight at the start of biofloc formation of 4.28 ± 0.13 g); T3, the
animals remained under the same conditions as the control until the
565
Aquaculture 479 (2017) 564–570
30th experimental day, followed by the initiation of biofloc formation,
with a mean weight of 11.10 ± 0.67 g; and T4, the animals were ex-
posed to the formation of microbial flocs only after 45 days of control
conditions, with a mean weight of 20.40 ± 1.09 g.
2.3. Maintenance of experimental conditions and formation of the biofloc
system
All tanks began the study with fresh water from an artesian well.
The water exchange was suspended as the formation of the biofloc
systems was initiated according to the previously established treat-
ments. In the control treatment and the treatments that had not yet
transitioned to the biofloc system, daily water exchange occurred at the
following rates: 50% until the 7th day; 75% from the 8th to the 22nd
day; 90% from the 23rd to the 30th day; 160% from the 31st to the
52nd day and 240% from the 53rd day to the end of the experimental
period. The water used for the exchanges was heated to a temperature
of 26–28 °C to avoid large variations in water temperature at the time of
exchange.
Fertilizations were performed for the formation and maintenance of
the bioflocs to stimulate the conversion of nitrogen into biomass by the
bacterial community. Molasses was used as the carbon source, and the
carbon content of this compound was considered to calculate the
amount added to the system. For the first three days, the method de-
scribed by Avnimelech (1999) was used. After this period, the carbon
input was determined according to Ebeling et al. (2006) employing a
C:N ratio of 6:1 based on the total ammonia nitrogen (TAN). Fertili-
zation was performed when the content was greater than or equal to
1 mg L− 1 of TAN.
2.4. Quality parameters for water and feed
The temperature, dissolved oxygen and pH were monitored daily in
the morning and evening (at 7:00 am and 4:00 pm, respectively) using
the multiparameter YSI 6920 V2® probe (Yellow Springs Incorporated -
YSI, Yellow Springs, OH, USA). The salinity was measured every three
days using the same multiparameter probe. The TAN and nitric nitrogen
(N-NO2−) were monitored daily and every two days, respectively. Total
ammonia was determined according to the methodology recommended
by UNESCO (1983), and nitrite was determined according to
Bendschneider and Robinson (1952). Samples were collected at
7:00 am before the water exchange for the control treatment and the
treatments that had not yet transitioned to biofloc systems. For pH
maintenance, calcium hydroxide (Ca(OH)2) was added when the values
were below 7.0 as recommended by Furtado et al. (2011).
Feed was offered ad libitum five times per day until the 45th day
when the schedule changed to four daily feedings until the end of the
experimental period. Commercial extruded feed for tilapia was used
with a 45% crude protein content (FRI-RIBE Nutreco, Brazil).
2.5. Animal performance and survival
Initially and then every fifteen days, biometric measurements were
taken using a reversible portable digital dynamometer (Instrutherm –
Instrumentos de Medição, Brazil). The performance was evaluated
based on zootechnical indices. The mean final weight (g) was calculated
by dividing the final biomass (g) by the final number of animals. The
final density (kg m− 3) was obtained by dividing the final biomass (kg)
by the volume in m3. The mean daily weight gain (g) was obtained by
dividing the individual biomass increment (mean final weight − mean
initial weight) by the number of days the animals remained in the ex-
periment. Feed conversion (FC) and the specific growth rate (SGR) were
obtained using the following formulas: FC = Feed intake/(final
weight − initial weight) and SGR (% day− 1) = ((ln final weight − ln
initial weight)/time) × 100. Simultaneous with the biometric mea-
surements, all animals were counted to determine survival as follows:
G.F. de Oliveira Alves et al. Aquaculture 479 (2017) 564–570

(final number of animals/initial number of animals) × 100.
2.6. Body composition
Two animals from each experimental unit were frozen in a freezer at
− 18 °C for analysis of body composition. The frozen animals were cut
into pieces approximately 1 cm in thickness, frozen at −40 °C for 48 h
and lyophilized for 72 h to eliminate moisture from the samples. After
lyophilization, the samples were disintegrated in a food processor until
the whole sample could pass through a 1 mm sieve.
The body composition was evaluated, determining the content of
dry matter, crude protein, ether extract, calcium, phosphorus and ash
according to the standard methodology of AOAC (2005). The energy
content was determined using a PARR 6200 bomb calorimeter (Parr
Instrument Company, IL, USA). The analyses were performed at the
Animal Nutrition Laboratory of the Veterinary School of UFMG.
2.7. Statistical analysis
The data obtained were subjected to analysis of variance taking into
account the necessary assumptions. When differences were found, the
means were compared using the Student-Newman-Keuls (SNK) test
(p < 0.05). The survival data were analyzed using the Kruskal-Wallis
non-parametric test (p < 0.05). The statistical analyses were per-
formed using the SAEG (2007) program.
3. Results
3.1. Water quality
The water physicochemical parameters differed (p < 0.05) among
the different transition times to the biofloc system (Table 1). The pH
was affected by the different transition times (p < 0.05). The higher
the time of residence in the biofloc system (i.e., the earlier the time of
transition to the system), the lower the pH values.
The mean TAN concentration followed a pattern similar to the pH,
with higher concentrations in the control group and lower concentra-
tions in the treatments that were under the biofloc system for a longer
period of time (p < 0.05) (Table 1). The time of formation of the
biofloc system significantly (p < 0.05) affected the nitric nitrogen (N-
NO2−) levels because there was a rapid elevation in and maintenance of
these levels for periods of 25–35 days, as shown in Fig. 1. The salinity
differed among treatments (p < 0.05), as shown in Table 1.
3.2. Animal performance
The mean values for the animal performance, mean final weight,
Table 1
Variables of water quality in juvenile Nile tilapia cropping (Oreochromis niloticus) trans-
ferred to the systems with biofloc conditions at different times of cultivation.
Variable Transfer moment
Control 1st day 15th day 30th day 45th day
final density, mean daily weight gain, survival, feed conversion and
specific growth rate parameters are shown in Table 2. The formation of
the biofloc systems at different time points when the animals were at
different weight ranges did not significantly affect the mean final
weight, survival and feed conversion (p > 0.05), as shown in Table 2.
The mean daily gain did not vary significantly (p > 0.05), although
an oscillation was observed in this parameter during the study due to
the transition of the animals from a system of daily water exchange to a
biofloc system where water exchange did not occur (Fig. 2).
The specific growth rate over time varied as a function of the
transition of the animals to the biofloc system in formation. However,
the mean specific growth rate was not affected (Table 2). The final
density (kg m− 3) was affected by the treatments (p < 0.05). A final
density of 17.44 kg m− 3 was recorded for the control; this value was
higher than the density in the experimental treatments, which did not
vary significantly among one another (p > 0.05). In the experimental
treatments, the final densities were 10.20, 12.47, 12.68 and
11.49 kg m− 3 for the T1, T2, T3 and T4 treatments, respectively. The
water consumption in the control was 6845.57 L kg− 1 and 112.34;
805.73; 1831.26; 4298.22 L kg− 1 in treatments T1, T2, T3 and T4,
respectively.
3.3. Body composition
The mean body composition and energy content values for the an-
imals transitioned to the biofloc system at different time points during
cultivation are summarized in Table 3. The dry matter content was not
affected by the transition of the animals to the biofloc system
(p > 0.05). The control group showed a higher crude protein con-
centration than did the other treatments (p < 0.05). Differences in the
protein content were not detected among the experimental treatments
transitioned at varying time points during cultivation to a biofloc
system (p > 0.05).
The ether extract content, which represents body fat, was affected
by the evaluated treatments. The treatment that transitioned to a bio-
floc system on the 1st day of the experiment (T1) had an ether extract
content of 223.61 g kg− 1, which was similar to treatments T2, T3 and
T4 but significantly different from the control treatment, which had an
ether extract content of 175.65 g kg− 1 in the body composition of the
animals. The mineral contents (ash) in the carcasses of animals transi-
tioned at different time points to the biofloc system were lower
(p < 0.05) than the contents measured in the control group animals.
When analyzed individually, the calcium and phosphorus levels showed
patterns similar to the patterns observed for the ash content, with the
highest values obtained in the control treatment and significant dif-
ferences compared to the other treatments (Table 3).
The gross energy results reflect the values obtained for both the
crude protein and the ether extract and ash. The gross energy value for
treatment T1, in which the animals were cultivated in a biofloc system
from day 1 of the experiment, was 22.69 kJ g− 1, which was higher than
that of the control (21.33 kJ g− 1); both values were equal to the other
treatments (Table 3).
CV (%)1
4. Discussion

Temperature (°C) 28.27 28.55 28.60 28.29 28.37 0.44 The water temperature did not differ among the evaluated treat-
Dissolved oxygen 5.71 5.70 5.59 5.67 5.69 2.03 ments and averaged 28.42 ± 2.41 °C, which was within the tempera-
(mg L− 1) ture range for the development, reproduction, and growth of tilapia of
pH 7.61a 7.05d 7.00d 7.28c 7.50b 0.93
TAN (mg L− 1)2 1.07a 0.41d 0.42d 0.54c 0.67b 6.56
20 to 35 °C (El-Sayed, 2006) and close to the range for growth opti-
N-NO2− 0.77b 16.93a 24.75a 20.80a 23.44a 16.79 mization of 29 to 31 °C (Popma and Lovshin, 1996). The dissolved
(mg L− 1)3 oxygen concentration remained above 5.0 mg L− 1 in all treatments,
Salinity (g L−) 0.11d 0.64a 0.64a 0.46b 0.24c 2.39 which was the minimum level recommended by Boyd and Tucker
(1998) to not limit the food intake, growth rate and general welfare of
Means followed by different letters in the row differ by the SNK test (p < 0.05).
1
Coefficient of variation.
the fish. In this study, the dissolved oxygen levels did not affect the
2
Total ammonia nitrogen. performance of the juvenile Nile tilapia or the development of the mi-
3
Nitrogen nitric. crobial community because the levels were maintained above those

566
G.F. de Oliveira Alves et al. Aquaculture 479 (2017) 564–570

Fig. 1. Changes in mean concentrations of nitric nitrogen (N-

70.00
NO2−) along the Nile tilapia cultivation (O. niloticus) according to
60.00 the different moments of transfer of fingerlings for biofloc system.
N-NO2- (mg L-1)

50.00
40.00
30.00
20.00
10.00
0.00
1 7 15 22 30 37 45 52 60 68 76
Time (days)

Control 1st day BFT (T1) 15th day BFT (T2)

30th day BFT (T3) 45th day BFT (T4)

Table 2 Table 3
Average final weight, final density, average daily gain, feed conversion, survival and Body composition and energy concentration (dry matter basis) in juvenile Nile tilapia (O.
specific growth rate in the production of juvenile Nile tilapia (Oreochromis niloticus) niloticus) transferred at different times of culture to the biofloc system.
transferred at different times to the biofloc system.
Variable Transfer moment CV (%)1
Variable Transfer moment CV (%)1
Control 1st day 15th day 30th day 45th day
Control 1st day 15th day 30th day 45th day
Dry matter (%) 23.38 24.81 24.23 23.54 23.40 5.48
Average body 50.30 44.15 46.51 47.00 42.86 8.79 Protein (g kg− 1) 629.60a 588.11b 596.20b 597.38b 594.26b 3.85
weight (g) Gross energy 21.33b 22.69a 21.85ab 21.96ab 21.60ab 4.37
Final density 17.44a 10.20b 12.47b 12.68b 11.49b 6.24 (kJ g− 1)
(kg m− 3) Calcium 46.11a 37.61b 40.44b 39.00b 42.15b 8.67
Average daily 0.64 0.56 0.59 0.60 0.55 9.05 (g kg− 1)
a b b b b
gain (g) Phosphorus 33.04 28.69 29.62 28.78 30.60 5.1
Survival (%)2 96.30 66.20 74.54 75.93 75.00 (g kg− 1)
Feed conversion 0.99 1.18 1.02 1.04 1.15 9.39 Ash (g kg− 1) 173.24a 146.05c 152.59bc 148.02bc 159.32b 6.3
Specific growth 4.75 4.55 4.59 4.65 4.52 2.91 Ether extract 175.65b 223.61a 204.21ab 202.74ab 195.37ab 13.07
rate (% (g kg− 1)
day− 1)
Means followed by different letters in the row differ by the SNK test (p < 0.05).
1
Means followed by different letters in the line differ through SNK test (p < 0.05). Coefficient of variation.
1
Coefficient of variation.
2
Análise through Kruskal Wallis test (p < 0.05).
was within the range considered suitable for tilapia (El-Sayed, 2006),
due to the corrections performed with calcium hydroxide (Ca(OH)2).
required by heterotrophic and nitrifying bacteria, as suggested by The decrease in the ammonia levels in the treatments transitioned to a
Timmons and Ebeling (2010). biofloc system compared to the control demonstrates the effectiveness
The decrease in pH observed in the treatments transitioned into a of raising the C:N ratio in maintaining adequate TAN levels in this
biofloc system might be associated with the demand for carbonate ions system (Avnimelech, 1999; Nootong et al., 2011; Samocha et al., 2007;
and bicarbonate by the microbial community for its development, Serra et al., 2015). The differences observed in the TAN concentrations
which can lead to the consumption of alkalinity and thus a reduction in between these treatments as a function of the time under the biofloc
pH levels (Ebeling et al., 2006). Despite the reduction in pH in the tanks system could be due to the stabilization phase the bacterial commu-
under the biofloc system, the pH remained at approximately 7.0, which nities were undergoing because the times of transition to the biofloc

Fig. 2. Changes in average daily gain, in-

1.40
dividual and every 15 days a juvenile Nile
tilapia (Oreochromis niloticus) that were
1.20
Average final weight (g)

transferred at different times of cultivation

to the biofloc system.
1.00

0.80

0.60

0.40

0.20

0.00
15 30 45 60 76
Time (days)
Control 1st day BFT (T1) 15th day BFT (T2)
30th day BFT (T3) 45th day BFT (T4)

567
G.F. de Oliveira Alves et al.
system were different. Widanarni et al. (2012) tested different storage
densities for tilapia (Oreochromis sp.) in biofloc systems and en-
countered similar results, varying from 0.01 to 1.13 mg L− 1 of TAN;
these results were lower than those obtained by Wambach (2013) in
Nile tilapia (O. niloticus) cultivations in biofloc systems under forma-
tion, where the mean values were between 3.32 and 4.48 mg L− 1 of
TAN.
The occurrence of nitrification has been observed in biofloc systems
(especially those under formation, such as in the present study), re-
sulting in nitrite accumulation (Burford and Longmore, 2001; Hari
et al., 2006; Azim and Little, 2008; Nootong et al., 2011; Widanarni
et al., 2012; Luo et al., 2014). Failure to perform water exchanges and
the addition of carbon sources, which raises the C:N ratio, may have
affected the nitrification rate (Ling and Chen, 2005), thereby con-
tributing to the nitric nitrogen accumulation and the maintenance of
high levels for up to seven weeks (Fig. 1). This result is similar to the
peaks reported during the formation of biofloc systems by various
studies (Hari et al., 2006; Nootong et al., 2011; Avnimelech, 2012; Luo
et al., 2014).
Systems operating with biofloc technology have minimum or zero
water exchange as a basic principle, which may lead to the accumula-
tion of nutrients from animal excretion and food remnants (Avnimelech
and Ritvo, 2003; Avnimelech, 2007). Because salt was not added to
increase the water salinity in this study, water exchanges were not
made in treatments under the biofloc system. Therefore, the differences
encountered in salinity (p < 0.05) are due to the accumulation of salts
derived from leftover feed. These differences might have occurred as a
function of the time maintained under a biofloc system because higher
levels of salinity were encountered in treatments that remained for
longer times under biofloc systems.
When animal performances were evaluated, we found that the fish
transitioned to the biofloc system did not show significant drops in
growth regardless of the timing of the transition. Survival analyzed
using the Kruskal-Wallis test was not affected (p > 0.05). When ana-
lyzing the time points during cultivation when mortalities occurred, we
found that the time points were concentrated immediately after the
reduction in the ammonia levels and the initial elevation in the N-NO2−
level, similar to the report by Day et al. (2016). However, the increase
in the N-NO2− level in the present study was not sufficient to affect
survival.
The adoption of high stocking densities in systems using biofloc
technology has been cited as one of its positive points (Avnimelech,
2012). The final density values for the treatments subjected to micro-
bial flocs in this study were close to those found by Widanarni et al.
(2012) in a study evaluating different stocking densities for red tilapia
(Oreochromis sp.), with a mean daily weight gain ranging from 0.52 to
1.16 g day− 1 for juveniles ranging from 78 to 190 g. Additionally, the
stocking density values were similar to the results obtained for the same
species by Wambach (2013), who performed a removal of solids during
cultivation. Crab et al. (2009) used the biofloc system in greenhouses
with water exchange to reduce the levels of nitrogen compounds and
achieved higher densities but recorded a lower mean daily weight gain
of 0.27 to 0.29 g day− 1. It is worth noting that the mean daily weight
gain in the present study reached almost twice that observed by Crab
et al. (2009), which could be associated with the effect of using lower
stocking densities and higher water temperatures compared to the cited
study.
At the time point when the animals were transitioned into biofloc
systems according to the pre-established treatments, a reduction in food
intake was observed similar to that reported by Day et al. (2016). This
momentary decrease in intake was primarily due to the occurrence of
N-NO2− peaks and the maintenance of these high concentrations for
weeks, which is characteristic of biofloc systems in formation
(Avnimelech, 2012; Nootong et al., 2011) and can be observed in Fig. 1.
After the reestablishment of feed intake (i.e., after the reduction in the
N-NO2− concentration), there was no significant difference in feed
568
Aquaculture 479 (2017) 564–570
conversion despite the higher feed intake in the treatments under the
biofloc system compared to the feed intake prior to the transitions
(Table 2). Feed conversion rates of 2.3 and 2.2 were reported for tilapia
hybrids (O. niloticus × O. aureus) cultivated from 125 to 415 g in a
biofloc system with a daily water exchange of 8% and in conventional
intensive systems (daily water exchange of 500%) by Milstein et al.
(2001). Azim and Little (2008) reported a feed conversion rate of 3.48
for juvenile tilapia (O. niloticus) in biofloc systems. Crab et al. (2009)
cited a conversion rate of 1.9 for juveniles ranging from 50 to 100 g,
Wambach (2013) reported an FC between 1.13 and 1.72 for juvenile
Nile tilapia cultivated at different stocking densities, and Day et al.
(2016) cited an FC of 1.01 for juveniles ranging between 36.2 and
80.0 g.
The comparison of these results is made difficult by the different
weight ranges of the animals studied, although the studies mentioned
above were also performed on animals in biofloc systems. Another
factor that completely differentiates the cultivation conditions in bio-
floc systems is the use of water from previous cultivations. This pre-
viously used water has pre-established microbial colonies that con-
tribute to the more rapid stabilization and maintenance of water quality
conditions and avoid the occurrence of peaks (especially nitric ni-
trogen), which is considered a positive point for this system
(Krummenauer et al., 2012).
Similar to the lack of an effect on feed conversion, the mean daily
weight gain and final mean weight did not differ, although there was a
momentary fluctuation in intake due to the increase in the nitric ni-
trogen levels, as shown in Table 2. Thus, compensatory growth may
have occurred in the present study. The hypothesis of the occurrence of
compensatory growth is supported by the variations in the specific
growth rate (SGR). After the initial formation of the biofloc system,
there was a drop in the SGR, followed by a recovery in the following
period independent of the time of transition. In contrast, the mean SGR
throughout the cultivation period did not differ according to the SNK
test (p > 0.05), as shown in Table 2.
Several studies have addressed the occurrence of compensatory
growth in fish production (Figueroa et al., 2000; Wang et al., 2000;
Takagi, 2001; Gao and Lee, 2012). Compensatory growth has been
defined as an accelerated growth phase after the reestablishment of
favorable conditions after passing through a period of growth depres-
sion caused by complete or partial food restriction. It can also be un-
derstood as an internal adjustment mechanism to improve the adapta-
tion of animals to different environments (Ali et al., 2003).
Quinton and Blake (1990) reported water quality as one factor that
might affect compensatory growth in fish. In the present study, the
variations in water quality parameters and the greater dynamism of this
type of system may have contributed to the occurrence of compensatory
growth when the animals transitioned to the biofloc system, as reported
by Avnimelech and Kochba (2009). Wang et al. (2000, 2005) observed
the occurrence of compensatory growth in tilapia hybrids grown both in
salt and freshwater when subjected to food restriction. The type of
compensation (partial or total) was determined according to the re-
striction period; when the food restriction lasted for up to one week, the
animals achieved total compensation, with higher intake and higher
specific growth rates than those of animals not under food restriction.
According to the authors, this finding indicated that the occurrence of
compensatory growth and hyperphagia were the main mechanisms
responsible for this phenomenon.
The determination of the duration of the food restriction period in
the present study was complicated because we did not aim to restrict
food from the animals. The reduction in intake is likely due to the
physiological adjustment needed to tolerate the increased levels of ni-
trogen compounds, especially N-NO2−. Additionally, feed was offered
daily, and the animals responded to the food and still had microbial
flocs in the cultivating unit. All of the treatments in which the animals
were transitioned to a biofloc system at some time point showed pos-
sible total compensatory growth because the final mean weights did not
G.F. de Oliveira Alves et al.
differ (p > 0.05). The ability to tolerate high concentrations of ni-
trogen compounds in water during the formation of the biofloc system
requires the use of species that can withstand variations in water
quality conditions (Boyd, 2004); thus, Nile tilapia are suitable for cul-
tivation in biofloc systems (Day et al., 2016). Based on the results
presented in this study, the formation of biofloc systems with finger-
lings from 1.4 g up to approximately 20 g did not affect the survival or
mean weight gain of the animals and also contributes to the reduction
in water consumption, providing savings of 98.36; 88.23; 73.25 and
37.21% in T1, T2, T3 and T4 treatments, respectively, in relation to the
control.
Few studies have addressed the body compositions of farmed fish
cultivated in biofloc systems. Azim and Little (2008) found no differ-
ences in the body compositions of animals produced in biofloc or re-
circulation systems; the animals in the biofloc systems showed the
following mean composition: 514.8 g kg− 1 of crude protein;
303.4 g kg− 1 of ether extract and 164.8 g kg− 1 of ash. However, the
energy content was 23.77 kJ g− 1, which differed from our observation
in this study where there was a distinction between the control group
and the other treatments.
The differences observed in this study may be due to compensatory
growth, which possibly occurred due to variation in the nitrite levels.
Jobling et al. (1994) suggested that the composition of weight gain
might differ between animals that underwent periods of restriction and
those that did not. The authors further stated that an improvement in
the feeding efficiency was to be expected when the increment was
preferentially lean mass compared to those that deposited larger
amounts of body fat. A result similar to the pattern described above was
observed for tilapia hybrids (O. niloticus × O. aureus) subjected to dif-
ferent feeding regimens, with restrictions of up to three days followed
by an equal period of re-feeding for a total of 60 days (Abdel-Hakim
et al., 2009). In contrast to these results, changes in carcass composition
were not detected in studies evaluating the compensatory growth in
tilapia hybrids after re-feeding and independent of the restriction time
periods (Wang et al., 2000, 2005) and improvements in the feeding
efficiency were observed.
Unlike reports from other studies, in the present study the animals
transitioned into the biofloc system showed changes in the body com-
position profile compared to the control group regardless of the time.
The transitioned animals appeared to have a lower protein content, and
the T1 treatment (transitioned on day 1) had a higher lipid content than
the control despite the lack of differences in feed conversion among the
treatments.
Ali et al. (2003) reported that body composition could vary over the
food restriction period and that general body composition after the re-
feeding and subsequent compensatory growth periods tended to be si-
milar to the body composition of the unrestricted animals. Thus, the
differences observed in the body composition profiles in the present
study may be more closely associated with the availability of microbial
flocs in the experimental units than with the compensatory growth it-
self. This discrepancy may be due to the possibility of the animals
consuming not only the supplementary diet but also the microbial flocs,
which may have complemented their diet.
For the marine shrimp Litopenaeus vannamei, the microbial biomass
can contribute between 18 and 29% of the nitrogen retained in the
carcass (Burford et al., 2004). For tilapia, microbial flocs may con-
tribute 25 to 50% of the protein requirement (Avnimelech, 2007;
Avnimelech and Kochba, 2009). Although these studies are associated
with the protein requirements of cultivated organisms, other composi-
tional aspects of microbial flocs may affect the body composition.
The deposition of lipids is often associated with the imbalance be-
tween energy and protein in the diets and the development phases of
the fish. Greater fat deposition in the body composition has been re-
ported in tilapia produced in biofloc systems (Long et al., 2015). In the
present study, the animals were in the early growth stage, and the
differences in body composition were likely due to nutritional aspects.
569
Aquaculture 479 (2017) 564–570
Because the C:N ratio is increased in biofloc systems for the assimilation
of nitrogen compounds by the microorganisms, the nutritional balance
between the diet and microbial flocs may not have been suitable for the
cultivated organisms, thereby affecting the body composition.
The accumulation of fat observed in animals transitioned on day 1
(T1) and kept for 76 days in the biofloc system was not desirable. If this
trend continues through the remaining stages of production up to the
point of slaughter, there may be a reduced carcass and fillet yield.
However, no differences were detected in the crude protein and ether
extract contents in the muscles of juvenile tilapia grown in biofloc
systems and those grown in recirculating systems (Luo et al., 2014).
Thus, there is a need to assess the production of Nile tilapia in biofloc
systems in all stages and under specific nutritional management stra-
tegies.
5. Conclusions
The performance of juvenile tilapia transitioned to biofloc systems
in formation and without water exchange suggested that the transition
could be performed with animals weighing 1.4 g. Thus, biofloc systems
can be initiated after the acquisition of masculinized fingerlings. The
use of alternatives to reduce or mitigate the effects of nitrogenous
compounds in biofloc systems (particularly nitrite) and the occurrence
of compensatory growth should be investigated in future studies.
Nutritional and feeding management strategies should be prepared for
the optimization of nutrition in animals cultivated in biofloc systems,
aiming at a higher accumulation of body protein and the avoidance of
excessive lipid deposition because biofloc systems are capable of al-
tering the body composition.
Acknowledgements
This research received support from PRPQ-UFMG (Pró-Reitoria de
Pesquisa, Universidade Federal de Minas Gerais, Brazil), FAPEMIG
(Fundação de Amparo à Pesquisa do Estado de Minas Gerais, Brazil),
CNPq (Conselho Nacional de Desenvolvimento Científico e
Tecnológico, Brazil) and CAPES (Coordenação de Aperfeiçoamento de
Pessoal de Nível Superior, Brazil).
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