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Aquaculture Research, 2017, 110 doi:10.1111/are.


Effect of inoculation of the cyanobacteria Oscillatoria

sp. on tilapia biofloc culture

Anselmo Miranda-Baeza1 opez1, Jos

, Mara de los A Mariscal-L opez-Elas2,
e A L
1 3
Martha E Rivas-Vega , Mauricio Emerenciano , Adolfo S anchez-Romero1 &
e L Esquer-M
Universidad Estatal de Sonora (UES) Unidad Navojoa, Navojoa, Sonora, Mexico
Departamento de Investigaciones Cientficas y Tecnol
ogicas de la Universidad de Sonora (DICTUS), Universidad de
Sonora, Hermosillo, Sonora, Mexico
~o Superior da Regi~
Universidade do Estado de Santa Catarina (UDESC) Centro de Educaca ao Sul, CERES, Laguna, Santa
Catarina, Brazil

Correspondence: A Miranda-Baeza, Universidad Estatal de Sonora (UES) Unidad Navojoa, Carretera a Huatabampo, km 5,
Navojoa, Sonora 85800, Mexico. E-mail:

expansion, including its good taste and fast

growth. Tilapia can be grown in ponds and cages,
Cyanophytes are the most ancient photosynthetic supports high stocking densities, resists adverse
organisms. During its evolution, they have devel- environmental conditions, tolerates low oxygen
oped various ecophysiological adaptation strategies concentrations and uses the primary productivity
to survive in extreme conditions. The environment of ponds as food source (El-Sayed 2006). In addi-
prevailing under biofloc cultures provides various tion to the biological and operational characteris-
conditions appropriate for cyanobacterial prolifera- tics that facilitate its growth, tilapia is widely
tion. An outdoor experiment (7 weeks) was per- accepted in the markets of both developed and
formed with a simple random design consisting of developing countries (FAO 2014).
four inoculation levels (by triplicate) of Oscillatoria One of the problems associated with tilapia
sp. (0.0, 0.1, 0.5 and 1 mg L1) in saltwater bio- farming in arid regions is the availability of fresh-
floc. The objective was to evaluate the effect of the water; therefore, its cultivation in salt water and
cyanobacteria inoculation on water quality and tila- biofloc could be one of the most viable alterna-
pia production parameters. The results indicated tives. Biofloc technology (BFT) allows operate with
that the amount of Oscillatoria sp. inoculated signifi- minimal water exchange and reduces the use of
cantly affected water quality (pH, chlorophyll a, TSS pelletized food. In a traditional culture, excess of
and NO3-N) and tilapia parameters (final weight, nutrients and organic matter must be discarded to
feed conversion ratio, specific growth rate and sur- prevent the increase in ammonia and other toxic
vival). No significant effects on dissolved oxygen, compounds, whereas biofloc systems take advan-
total ammonia nitrogen (TAN) or NO2-N were tage of these components through stimulation of
observed. We recommend identifying the cyanobac- microbial biomass, which is used as a live and per-
teria species that are able to grow in a biofloc sys- manent source of food (Ekasari, Crab & Verstraete
tem and their possible adverse effects on the system. 2010; L opez-Elas, Moreno-Arias, Miranda-Baeza,
Martnez-Cordova, Rivas-Vega & M arquez-Ros
Keywords: biofloc, cyanobacteria, nitrogenous 2015).
compounds, Oscillatoria sp., tilapia culture In biofloc systems, a highly diverse consortium
of unicellular organisms is present including auto-
trophs, heterotrophs, prokaryotes and eukaryotes
(Martnez-Cordova, Martnez-Porchas, Emeren-
Tilapia is one of the most cultivated fish in the ciano, Miranda-Baeza & Gollas-Galv an 2016). Sev-
world. Several features have favoured their eral authors have reported the presence of

2017 John Wiley & Sons Ltd 1

Oscillatoria sp. on biofloc tilapia culture A Miranda-Baeza et al. Aquaculture Research, 2017, 110

cyanobacteria in biofloc cultures (Ju, Forster, Con- Oscillatoria, cyclic peptides and toxic alkaloids have
quest, Dominy, Kuo & Horgen 2008; Ballester, been identified (Sivonen & Jones 1999).
Abreu, Cavalli, Emerenciano, De Abreu & Wasie- According to De Schryver, Crab, Defoirdt, Boon
lesky 2010; Becerra-D orame, Martnez-Cordova, and Verstraete (2008) and Martnez-C ordova et al.
Lopez-Elas & Martnez-Porchas 2011). However, (2016), biofloc systems convert excess nutrients in
the impact that a high concentration of these the microbial biomass. These nutrients are subse-
micro-organisms may have on water quality and quently consumed by the cultured animals,
productive parameters has not been assessed. To increasing the water residence time (Ekasari et al.
know this, information is important, specially 2010), and reducing costs from water
because the cyanobacteria can grow under hetero- consumption.
trophic conditions, fix N2 and generate Avnimelech (2007) evaluated the assimilation
cyanotoxins. of biofloc by tilapia and concluded that its compo-
The genus Oscillatoria belongs to the class Oscil- nents can contribute to approximately 50% of the
latoriales and division Cyanobacteria and is char- protein requirement for tilapia. Azim and Little
acterized by long filamentous flattened cells (2008) evaluated tilapia (Oreochromis niloticus) cul-
without a dark green mucilaginous sheath. It is tured in biofloc and determined that fish produc-
very common in microbial mats in both marine tion was 45% increased in the experimental tanks
and freshwater environments, especially in the compared with the control (traditional culture).
presence of high organic load (Fuentemayor, Jonte, However, to contribute to the qualitative nutri-
Rosales-Loaiza & Morales 2009). ent requirements and obtain good growth rates, it
Cyanobacteria are photosynthetic micro-organ- is necessary that the biofloc be ingested, digested
isms with great morphological and structural vari- and assimilated. The environment that develops
ability; throughout evolution, they have developed under biofloc cultures provides various conditions
various ecophysiological adaptive strategies in appropriate for cyanobacterial proliferation. The
response to adverse environmental conditions risk of pollution by an aggressive cyanobacterial
(Lopez-Rodas, Maneiro & Costas 2006). As a strain could occur during fish farming. Thus, it is
result, cyanobacteria can be found in places with necessary to generate information about the effects
extreme temperature, humidity, solar radiation, of the presence of cyanobacteria in BFT. The main
pH and nutrient concentrations. objective of this study was to evaluate the effect of
Cyanobacteria are opportunistic micro-organ- Oscillatoria sp. inoculation at four concentrations
isms. Specific conditions, such as nutrient avail- on nitrogenous compounds, suspended solids and
ability, temperature and light intensity, can the production parameters of tilapia (Oreochromis
promote the explosive growth of their populations niloticus 9 Oreochromis mossambicus) reared in
(Carpenter & McCarthy 1975). In addition, the saltwater biofloc system.
ability of cyanobacteria to adapt to growth in a
wide range of radiation levels is regulated by a
Materials and methods
complex system of thylakoid components (Mura-
kami & Yoshihiko 1991).
Isolation and maintenance of the strain
Heterocysts in cyanobacterial cells are almost
colourless, have a thick wall and perform the func- The cyanobacterial strain was obtained from the
tion of molecular nitrogen fixation (N2), which bottom of shrimp ponds in Sonora State (north-
can occur under anaerobic conditions (Neuer, west Mexico), isolated by serial dilution as
Papen & Bothe 1983). In addition to species with described by Band (2007) and subsequently main-
heterocysts, other species without these cells may tained under laboratory conditions until the exper-
be able to fix N2, including the multicellular gen- iments began. Identification at the genus level was
era Oscillatoria (Carpenter & McCarthy 1975) and conducted by the Institute of Marine Sciences and
Microcoleus (Malin & Pearson 1988). Limnology of the Universidad Nacional Aut onoma
Cyanobacteria are micro-organisms associated de Mexico. The F/2 (Guillard & Ryther 1962) cul-
with toxin production. Cyanotoxins are a diverse ture medium was used during strain maintenance.
group of toxins with complex chemical structure Light was provided by white lamps, and the tem-
and varied toxicological effects. The type of toxin perature was maintained between 22 and 25C.
produced depends on the cyanobacterial genus. In The strain was transferred every 15 days.

2 2017 John Wiley & Sons Ltd, Aquaculture Research, 110

Aquaculture Research, 2017, 110 Oscillatoria sp. on biofloc tilapia culture A Miranda-Baeza et al.

following the suggestion of Furtado, Poersch and

Experimental design
Wasielesky (2011).
This experiment was a simple random design with The chlorophyll a was measured weekly, and
four treatments by triplicate. The treatments con- 10 mL of samples was obtained and concentrated
sisted of inoculation with 0.0, 0.1, 0.5 and in a refrigerated centrifuge (10 000 rpm for 5 min
1 mg L1 Oscillatoria sp. corresponding to treat- and 4C). The supernatant was discarded, and
ments C0, C1, C2 and C3 respectively. 9 mL of 90% acetone was added following the
The 12 experimental units (plastic tanks with a technique of Strickland and Parsons (1972). Total
capacity of 70 L) were filled with 50 L of salt suspended solids were measured as outlined in
water (35 &) previously sterilized with calcium Hach Method 8006 (Hach 2007), using a Hach
hypochlorite (3 ppt) and neutralized with sodium DR/2800 spectrophotometer (Hach Co. Loveland,
thiosulfate. Each tank was stocked with 250 g of CO, USA). The method was previously calibrated
hybrid red tilapia juveniles (Oreochromis mossambi- and corrected by the traditional gravimetric
cus x Oreochromis niloticus), equivalent to method according to Miranda-Baeza, Voltolina and
5 kg m3. The organisms (14.5  2.6 g) were Cordero-Esquivel (2006).
randomly selected from a pre-breeding tank. Dur- Total ammonia nitrogen (TAN), nitrite nitrogen
ing the experiment, the tilapia were fed to satia- (NO2-N) and nitrate nitrogen (NO3-N) were mea-
tion with commercial diet with 35% protein sured every week. For this purpose, water samples
(Purina; Agribrands Purina Mexico, S.A. de C.V. (250 mL) were collected and centrifuged, and the
Estado de Mexico, MEX). supernatant was used to measure the respective
To supply oxygen and maintain solids in sus- nutrients. The TAN was measured with the salicy-
pension, constant aeration was provided by an late method (8155), NO2-N by the diazotization
electric blower of 1/3 HP and air stones placed on with ferrous sulphate in acid medium method
the bottom. The study was carried out in outdoor (8507) and NO3-N by the cadmium reduction
conditions with a natural photoperiod (14-h light; method (8171) according to the procedures
10-h dark) and no temperature control. No water described in the Hach spectrophotometer manual
exchange was performed, but evaporation losses (Hach 2007).
were compensated by freshwater. The carbon/ni-
trogen (C/N) ratio was maintained at 1:15, and
Tilapia production parameters
the input of C and N due to food was considered
and complemented with glucose (unrefined sugar) The specific growth rate (SGR) of the tilapia was
according to Avnimelech (2009). The growth per- estimated from the equation suggested by El-Har-
iod lasted 7 weeks. To maintain the total sus- oun, Goda and Chowdhury (2006)
pended solids below to 400 mg L1, 50% of tank
volume was filtered through 10-lm mesh on days SGR Ln final weightg
20 and 35.  Ln initial weightg100=culture time:

The survival rate was obtained from the rela-

Water variables measurement
tionship (final number of organisms/initial num-
Temperature and dissolved oxygen were mea- ber) 9 100.
sured twice daily (8:00 and 16:00 hours) using
YSI 58 digital oximeter (YSI Inc., Yellow Springs,
Data analysis
OH, USA). The pH was measured daily with a
Denver Instruments pH-10 digital pH meter The assumptions of normality and homogeneity of
(Denver Instruments, Denver, CO, USA), and variance were verified. Furthermore, one-way
salinity was measured weekly with an Aquatic ANOVA (Zar 1996) was performed to determine sig-

Eco-Systems refractometer (Pentair Aquatic Eco- nificant differences between the basic water vari-
Systems, FL, USA). Total alkalinity was measured ables and tilapia production parameters. In cases
on a weekly basis according to APHA (1998). where the differences were significant (P < 0.05),
Corrections of alkalinity (as CaCO3) were per- a posteriori multiple comparison Tukeys tests
formed to maintain concentrations above were applied. STATISTICA for Windows 5.1, StatSoft
100 mg L1 using bicarbonate (NaHCO3) Inc. software was used for data processing.

2017 John Wiley & Sons Ltd, Aquaculture Research, 110 3

Oscillatoria sp. on biofloc tilapia culture A Miranda-Baeza et al. Aquaculture Research, 2017, 110

Results 1000

Chlorophyll a (g L1)
The average temperature throughout the study 800

ranged between 25.3 and 25.8C in the morning 600

and between 27.9 and 28.6C in the afternoon.
No significant differences among treatments were 400
y = 914.95x2+ 1394.2x + 293.28
R = 0.924
observed (Table 1). Dissolved oxygen ranged from 200
4.29 to 4.47 mg L1 in the morning and from
3.54 to 3.80 mg L1 in the afternoon without sig- 0
0 0.2 0.4 0.6 0.8 1 1.2
nificant differences between treatments. The pH Oscillatoria inoculation (mg L1)
presented means between 7.70 and 7.81 in the
Figure 1 Relationship among inoculation level of
morning and 7.62 and 7.71 in the afternoon. In Oscillatoria (0.0, 0.1, 0.5 and 1 mg L1) and chloro-
both cases, the difference between treatments was phyll a (global averages for each treatment) into the
significant (P < 0.05). The salinity varied from tilapia culture tanks.
34.9 to 35.7& without a significant difference
among treatments.
The overall average of chlorophyll a ranged Table 2 Chlorophyll a total suspended solids and
from 226 to 780 lg L1. The minimum corre- alkalinity (mean  SD) in the different treatments
sponded to treatment C0 and the maximum to
Chlorophyll a TSS Total alkalinity
treatment C3. The ANOVA revealed significant differ-
Treatment (lg L1) (mg L1) (mg CaCO3 L1)
ences (P < 0.05) between treatments (Table 3).
The averages of chlorophyll a versus the inocula- C0 226  75a 277  48a 108.2  7.2
C1 517  120b 258  57a 114.1  8.4
tion level were described by a polynomial model
C2 728  117c 288  97ab 112.4  10.2
(Fig. 1). The average total suspended solids varied C3 780  98c 337  102b 105.3  12.1
from 258 to 337 mg L1. The minimum corre-
sponded to treatment C1 the maximum to C3, and One-way ANOVA. Different letters in the same column indicate
significant differences, P < 0.05; a < b < c.
significant differences were identified (P < 0.05)
(Table 2). The total alkalinity (as CaCO3) varied
from 105.3 to 114.1 mg L1, with no significant Throughout the experiment, the value ranged
differences among treatments (P < 0.05) (Table 2). between 0.82 and 0.85 mg L1 without a signifi-
The TAN was not detected during day 1. How- cant difference (P < 0.05).
ever, TAN values ranged between 3 and 5 mg L1 The NO3-N increased from day 9, peaked at day
on day 9. Subsequently, the concentration 18 (55 and 65 mg L1) and ended between 10
decreased significantly and was close to zero at and 20 mg L1 (Fig. 2c). The average throughout
day 18 (Fig. 2a). The overall average for the entire the experiment ranged between 22 and
experiment ranged between 0.96 and 33 mg L1. The lowest concentration corresponded
1.75 mg L1 with no significant differences to treatment C3, and the highest concentration
between treatments (P < 0.05). was found with treatment C0. The difference was
The NO2-N increased from day 6 and peaked significant (P < 0.05).
around day 12. At the end of the experiment, the The initial biomass of tilapia varied from 5.0 to
concentration reached 0.01 mg L1 (Fig. 2b). 5.1 kg m3. At the end of the study, the average

Table 1 Temperature, dissolved oxygen and pH (mean  SD) in the different treatments

Temperature (C) Oxygen (mg L1) pH

Tr am pm am pm am pm

C0 25.8  2.0a 27.9  1.4a 4.30  0.28a 3.80  0.26a 7.70  0.02a 7.62  0.02a
C1 25.8  2.4a 28.6  1.6a 4.47  0.28a 3.78  0.20a 7.80  0.06b 7.71  0.04b
C2 25.7  2.2a 28.1  1.7a 4.39  0.39a 3.73  0.39a 7.81  0.05b 7.71  0.03b
C3 25.3  2.3a 28.1  1.6a 4.29  0.35a 3.54  0.42a 7.74  0.04ab 7.68  0.06ab

One-way ANOVA. Different letters in the same column indicate significant differences, P < 0.05, a < b.

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Aquaculture Research, 2017, 110 Oscillatoria sp. on biofloc tilapia culture A Miranda-Baeza et al.

(a) 6 Discussion
5 C1 Temperature and salinity were maintained within
C2 the suitable ranges reported by El-Sayed (2006)
TAN (mg L1)

C3 for growing tilapia in seawater. In biofloc systems,
dissolved oxygen (DO) is considered one of the crit-
ical variables due to the high respiratory rate of
1 micro-organisms. In such systems, the cultured
0 species consumes a reduced proportion of the DO
0 10 20 30 40 50
(b) (Ray, Seaborn, Leffler, Wilde, Lawson & Browdy
C0 2010; Vinatea, G alvez, Browdy, Stokes, Venero,
C1 Haveman, Lewis, Lawson, Shuler & Leffler 2010),
NO2-N (mg L1)

C2 and the remainder is used by the micro-organisms

C3 present: the bacterial population, which consumes
oxygen continuously (Avnimelech 2009), and
0.5 microalgae, which consume oxygen during the
night or on cloudy days.
0.0 In this study, the overall mean of dissolved oxy-
0 10 20 30 40 50
(c) gen was similar between treatments. However, in
80 C0 the afternoon, the concentration was reduced in
70 C1 treatments with more Oscillatoria inoculum. In
NO3-N (mg L1)

contrast to expectations, the morning readings
40 exhibited higher levels of DO, which indicates that
30 the production of oxygen by photosynthesis was
20 unable to compensate the loss of DO due to
10 decreased solubility caused by afternoon tempera-
0 ture. An increase of 3C (the difference between
0 10 20 30 40 50
Culture day morning and afternoon) provoked a decreased of
1 mg L1 DO. Therefore, the high content of
Figure 2 Average values of total ammonia nitrogen, micro-organisms in the biofloc may increase the
nitrite nitrogen and nitrate nitrogen (mg L1) during
risk of hypoxia when aeration is not efficient.
the experiment.
However, most tilapia species can tolerate low DO
levels; Teichert-Coddington and Green (1993)
ranged from 6.0 to 8.2 kg m3 with significant dif- found that reducing aeration in tilapia ponds from
ferences (P < 0.05); the lowest yields were obtained 30% to 10% of oxygen saturation did not affect
in the C2 and C3 treatments, and the highest were fish growth. In this study, the overall DO averages
noted in C1 and C0. The final individual weight were between 3.54 and 4.29 mg L1, which is
varied from 21.6 to 24.1 g with significant differ- appropriate for tilapia culture (El-Sayed 2006).
ences between treatments (P < 0.05). The lowest Additionally, the levels were into the interval
weight was obtained in the C2 and C3 treatments, (3.07.5 mg L1) reported by Azim and Little
whereas the highest was in C1 and C0 (Table 3). (2008) who reared tilapia in biofloc system.
The feed conversion ratio (FCR) varied from Another important variable in biofloc systems is
1.35 to 1.85 with significant differences between pH, due to its effect on the ionization of ammonia.
treatments (P < 0.05). The lowest averages were Higher values (pH >9) reduce this ionization and
obtained in C0 and C1, and the highest averages cause a considerable proportion of NH4+ to
were observed in C2 and C3. Survival ranged from become NH3, which even at low concentrations is
72% to 98%; the lowest corresponded to C3 and toxic to many aquatic species (Hopkins, Hamilton,
the highest to C0, with significant differences Sandier, Browdy & Stokes 1993). Biological factors
between treatments (P < 0.05). Specific growth that influence pH include photosynthesis and res-
also showed significant differences (P < 0.05); the piration, both of which have great activity in bio-
lowest average (0.36) was obtained in C3 and the floc systems. Photosynthesis can have a noticeable
highest (1.05) in C0 (Table 3). effect on the pH of water. Photosynthetic activity

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Oscillatoria sp. on biofloc tilapia culture A Miranda-Baeza et al. Aquaculture Research, 2017, 110

Table 3 Tilapia production parameters (mean  SD) in the different treatments


Treatment Initial (kg m3) Final (kg m3) Final weight (g) FCR SGR Survival (%)

a b b a b
C0 5.0  0.03 8.22  0.48 24.0  1.7 1.35  0.12 1.05  0.14 98  0.5b
C1 5.1  0.15a 8.12  0.23b 24.1  1.0b 1.40  0.18a 0.97  0.12b 96  2.6b
C2 5.0  0.05a 6.26  0.15a 21.7  0.6a 1.83  0.14b 0.45  0.06a 82  3.6a
C3 5.1  0.05a 6.00  0.34a 21.6  0.5a 1.85  0.08b 0.36  0.13a 72  3.0a

One-way ANOVA. Different letters in the same column indicate significant differences, P < 0.05, a < b.

removes CO2 from the system and causes an addition, they can be grown in fully hetero-
increase in the pH of the water. In contrast, respi- trophic conditions (Gallon, Hasem & Chaplin
ration provides CO2, and the pH decreases (Tim- 1991). This phenomenon occurred in this experi-
mons, Ebeling, Wheaton, Summerfelt & Vinci ment; although treatment C0 was not inoculated
2002). Lower pH levels were recorded in treat- with Oscillatoria, their presence became observ-
ment C0, whereas it was the highest in the C1C3 able after 10 days of culturing and increased
treatments. These results were consistent with moderately.
expectations, as C0 had the lowest photosynthesis In the present experiment, chlorophyll a global
rate. means ranged from 226 to 780 lg L1. The high-
In aquaculture ponds, carbon dioxide, pH, alka- est concentration was noted in treatment C3 and
linity and hardness are interrelated. The water was consistent with the range of 500 to
with moderate-to-high alkalinity and slightly basic 900 mg L1 reported by Decamp, Conquest, Cody,
pH (7.08.3) has good buffering capacity; biofloc Forster and Tacon (2007) for shrimp farming with
systems lose this ability and therefore require fre- zero water exchange. Figure 1 describes the effect
quent restoration (Azim & Little 2008). Furtado of inoculation over chlorophyll a concentration
et al. (2011) suggest alkalinity greater than and indicates that the Oscillatoria inoculum effect
100 mg L1 and pH above to 7 for saltwater bio- is nonlinear, and the lower initial inoculum of
floc systems; in this study, alkalinity was con- 0.1 mg L1 generated an average concentration of
trolled; as a result, no significant differences 517 lg L1 of chlorophyll a, while with 1 mg L1
among treatments were determined. It was not was of 780 lg L1.
necessary to restore pH because its levels were Although several authors have reported the
greater than 7. abundance of cyanobacteria in biofloc as
Several authors have reported the presence of cells mL1, it is clear that errors potentially
cyanobacteria in biofloc cultures. In a study with occurred during the quantification of filamentous
post-larval shrimp, Becerra-D orame et al. (2011) forms, because these organisms are entangled in
reported cyanobacteria concentrations of the bioflocs. In addition, fragmentation occurs
2.1 9 104 cells mL1 in a heterotrophic biofloc during any separation process. Thus, in this
and 3.3 9 106 in an autotrophic biofloc. These study, we decided to perform indirect determina-
results demonstrate the ability of cyanobacteria to tion through chlorophyll a. Fuentemayor et al.
proliferate in biofloc systems. In another study, (2009) conducted an Oscillatoria sp. culture
Ballester et al. (2010) reported that filamentous under controlled conditions and estimated that
cyanobacteria reached 218 filaments mL1 and for each mg of biomass, between 2.10 and
17 200 cells mL1 in a biofloc system. Further- 2.97 lg L1 of chlorophyll a was extracted.
more, Ju et al. (2008) indicated that cyanobacteria Because the water was sterilized, it is possible to
accounted for 5.9% of the total phytoplankton assume that the chlorophyll a detected in this
cells in a biofloc shrimp culture, whereas chloro- study came from Oscillatoria; at the end of the
phytes reached 82.3%. culture, we obtained cyanobacterial biomass
The presence of cyanobacteria in BFT is ranging from 125 mg L1 (C0) to 260 mg L1
explained by their ability to adapt to adverse (C1, C2 and C3), corresponding to slightly
conditions of light, temperature and high organic greater than 50% of TSS in the C1, C2 and C3
matter load (Fuentemayor et al. 2009); in treatments.

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Aquaculture Research, 2017, 110 Oscillatoria sp. on biofloc tilapia culture A Miranda-Baeza et al.

The presence of TSS is another major variable day 15 of the experiment. The gap between the
in biofloc systems. Azim and Little (2008) reported TAN, NO2-N and NO3-N curves coincides with the
values of TSS from 560 to 597 mg L1 in a cul- appearance of ammonia-oxidizing bacteria (Nitro-
ture of tilapia in biofloc, whereas Avnimelech somonas) and subsequently nitrite-oxidizing bacte-
(2007) reported a range of 460 to 643 mg L1. In ria (Nitrobacter). This process has been widely
the present study, the average was between 277 documented in recirculating water systems (Tim-
and 337 mg L1, which is lower than those mons et al. 2002).
reported in the papers cited. We decided to main- In this study, the nitrate concentration showed
tain TSS below 400 mg L1 by conducting partial an increase from day 1 to day 16, with a later
biofloc harvests to prevent clogging of gills and decrease to the end of the experiment. The outdoor
risks due to low oxygen diffusion from the high biofloc cultures do not have light-limited condi-
concentration of Oscillatoria sp. in the C1, C2 and tions, and it is possible to obtain high autotrophic
C3 treatments. Emerenciano, Gaxiola and Cuzon production. This condition and the moderate C/N
(2012) indicate that high levels of TSS can affect ratio (1:15) used in our study caused high
the oxygen diffusion through the gills, but we did demand of nutrients; therefore, it is possible that
not found specific information to determine the Oscillatoria had consumed NO3-N when TAN was
exact effect of biofloc biomass over the oxygen dif- scarce (Fig. 2); Azim and Little (2008) as well as
fusion in tilapia gills; it could be an interesting Ekasari and Maryam (2012) reported in biofloc
topic for further research. tilapia cultures. It has been documented the
At the beginning of the experiment, the TSS microalgae metabolism is capable to use different
level in all treatments was 100 mg L1 with a sources of inorganic nitrogen (Li, Horsman, Wang,
rapid increase. Therefore, it was necessary to crop Wu & Lan 2008).
biofloc on days 20 and 35 (50% of volume was fil- The presence of microalgae may have an advan-
tered through 10-lm mesh). The C3 treatment tage in biofloc cultures because they permit nitro-
recorded the highest increase (reaching 570 mg gen compounds to re-enter into the food chain.
L1 at day 20). For treatment C0, although it did However, at high concentrations, microalgae could
not receive inoculum, the amount of TSS was sim- be a risk because they can become consumers of
ilar to that in C1 and C2, indicating that the rate O2 on cloudy days. In microalgae-dominated sys-
of formation and accumulation of SST in an auto- tems, the maximum NH4+ assimilation capacity is
trophic system may be similar to that in a mixo- 0.7 g m2 day1. In contrast, in heterotrophic
trophic system. bacteria-dominated systems, the assimilation
The different inoculation levels of Oscillatoria sp. capacity can be unlimited if an appropriate C/N
showed no significant differences in TAN concen- ratio is maintained (Avnimelech 2009).
tration. However, during the first 15 days, in the The cyanobacterium Oscillatoria sp. had signifi-
C3 treatment, the concentration remained lower cant effects on the tilapia biomass produced. Lower
than that in the other treatments. Some microal- production was recorded in treatments C2 and C3,
gae prefer NH4+ over other chemical forms of N. whereas similar production was noted in treat-
Hii, Soo, Chuah, Mohd-Azmi and Abol-Munafi ments C0 and C1. The final average weight in
(2011) conducted a study with Nannochloropsis sp. treatments C2 and C3 was between 21.6 and
In the presence of ammonium and nitrate, the 21.7 g, whereas the final average was 24 g in
microalgae had a preference for NH4+ and also treatments C0 and C1.
exhibited increased growth when this compound The incorporation of Oscillatoria had a signifi-
was the unique source of N. These phenomena are cant effect on survival. Treatment C0 had 98%
consistent with our study because the use of NO3 survival, whereas survival was only 72% in treat-
was initiated when TAN was consumed (day 15) ment C3. The genus Oscillatoria may produce toxic
(Fig. 2). cyclic peptides, including microcystins, anatoxin-a
The maximum level of NO3-N was achieved on and aplysiatoxins (Sivonen & Jones 1999).
day 20; afterwards, it began to decline along with Although the toxicity of Oscillatoria in tilapia has
NO2-N. In the case of C1, C2 and C3, the decrease not been studied, the results of this experiment
in NO3-N can be explained because Oscillatoria indicate that its presence caused negative effects
started using oxidized N compounds when the on the growth performance of tilapia, especially in
TAN had reached low levels (below 1 mg L1) at treatments with greater abundance (C2 and C3).

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Oscillatoria sp. on biofloc tilapia culture A Miranda-Baeza et al. Aquaculture Research, 2017, 110

More research is needed to evaluate the pres- Avnimelech Y. (2007) Feeding with microbial flocs by
ence of cyanobacteria in biofloc systems consider- tilapia in minimal discharge bioflocs technology ponds.
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conversion ratios (1.35 and 1.40), which exhibited formance of Farfantepenaeus paulensis juveniles nursed
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FCR values obtained in the C0 and C1 treatments CIBNOR, Mexico.
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