Aquacultural Engineering 82 (2018) 56–62

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Aquacultural Engineering
journal homepage: www.elsevier.com/locate/aque

Synthetic fibre as biological support in freshwater recirculating aquaculture T

systems (RAS)
⁎
Marco Shizuo Owataria, , Gabriel Fernandes Alves Jesusa,
Marcos Estevão Santiago de Melo Filhob, Katt Regina Lapac, Maurício Laterça Martinsa,
José Luiz Pedreira Mouriñoa
a
AQUOS-Aquatic Organisms Health Laboratory, Aquaculture Department, Federal University of Santa Catarina (CCA, UFSC), Rodovia Admar Gonzaga 1346, 88040-
900, Florianópolis, SC, Brazil
b
Chemical Engineering Department, Federal University of Santa Catarina, Brazil
c
Aquaculture Department, Federal University of Santa Catarina, Brazil

A R T I C LE I N FO A B S T R A C T

Keywords: The aim of this study was to evaluate the use of synthetic fibre as a biological support for the adhesion of
Rhamdia quelen nitrifying bacteria in an aquaculture recirculation system (RAS). It was developed from three assays over 120
Oreochromis niloticus days. In the first assay, the synthetic fibres used as biological support were introduced in tanks of biological
Intensive farming filtration of the system for posterior respirometry analysis and scanning electron microscopy (SEM).
Biological filter
Respirometry and SEM were performed 10 days after inoculation with nitrifying microorganisms. Water quality
Nitrifying bacteria
parameters were monitored daily, and the respirometry showed that the bacteria in this assay were consuming
the following amounts of ammonium: concentrations [C1]35.369 mg NH3/L, R2 = 0.9912; [C2]51.628 mg NH3/
L, R2 = 0.9883; [C3]79.494 mg NH3/L, R2 = 0.986; and [C4]215.225 mg NH3L, R2 = 0.9934. In the second
assay, a 1920-L tank was stocked with 120 Nile tilapia, Oreochromis niloticus, with an initial weight
32.11 ± 7.6 g and a biomass of 3.8 kg. After 60 days, the tank and its contents were assessed to evaluate
zootechnical parameters and physical–chemical parameters of water quality. From these results, a third assay
was developed in which the biomass of fish was increased to challenge the recirculation system. The tank was
stocked with 480 jundiá Rhamdia quelen (initial weight 11.34 ± 2.4 g and biomass 5.4 kg) for 60 days. In both
the tilapia and jundiá assays, the fish were fed four times per day with a commercial diet of 35% crude protein
and 42% crude protein, respectively, at 5% of each individual fish’s body weight. At the end of the zootechnical
assays, the synthetic fibres used showed efficient biological support for bacterial growth, as confirmed by
scanning electron microscopy. The fibres also demonstrated maintenance of the water quality, which allowed
good fish growth in the recirculating aquaculture system, and the maintenance of up to 11.19 kg/m³ of biomass
of fish.

1. Introduction processes, whereby bacteria, under good environmental conditions,

The recirculation aquaculture system (RAS) is based on high fish
densities and can use 99% less water than traditional cropping systems.
It is costly to execute a RAS, and any mistake in the dimensions or
choice of equipment can result in damage to the system. Nevertheless,
when well built, a, RAS provides great advantages and allows for the
control of physical–chemical water variables in culture (Nazar et al.,
2013). An RAS can be designed to remove undesirable residues in the
water, such as sand filters, sedimentation structures and biofilters (Bijo,
2007).
Removal of ammonia in an RAS is accomplished by nitrifying
oxidise ammonia to create nitrite. Nitrifying bacteria are mostly che-
moautotrophic, and they generate their energy from the oxidation of
ammonia and nitrite (Liu et al., 2013; Sliekers et al., 2002), known as
epibacterial community, which are needed to support substrate de-
gradation.
The determination of the maximum and minimum nitrification ca-
pacities of a biofilter is directly linked to its biofilm. This biofilm is
formed on the surface area available for adhesion of layers of different
populations and associations of microorganisms in the support material.
Media materials, also called filter media, are commonly evaluated and
compared across the total surface area available per cubic meter of

⁎
Corresponding author.
E-mail address: marco.owatari@posgrad.ufsc.br (M.S. Owatari).

https://doi.org/10.1016/j.aquaeng.2018.06.001
Received 29 January 2018; Received in revised form 26 April 2018; Accepted 11 June 2018
Available online 25 June 2018
0144-8609/ © 2018 Elsevier B.V. All rights reserved.
M.S. Owatari et al.
media. However, estimating the rates of nitrification based on the
theoretical surface area can often be misleading because the biofilm
that covers the media is able to modify the characteristics of the media
and create a new surface area, reducing the area available to nitrifying
bacteria (Guerdat et al., 2010). From this perspective, the efficiency of
the biological filter and the nitrification processes in an RAS are directly
related to the types of media used as biological support for bacterial
adhesion (Lekang and Kleppe, 2000). A variety of both natural and
artificial media can be used in the biological filters of an RAS, and they
must favour the growth and adhesion of nitrifying bacteria (Ridha and
Cruz, 2001).
Microbiological characteristics of bacterial biomass that act in the
nitrification process and the associated activity of microorganisms may
be determined using a kinetic study that involves multiple methods.
Microbiological characteristics are based on variations in the substrate
consumption and formation of some products in the environment,
which are directly related to biological reactions, dissolved oxygen
variations, pH, temperature and alkalinity (Timmons and Ebeling,
2010). Respirometry stands out as a rapid and simple technique that
can be used to determine the velocity of the biological consumption of
oxygen under experimental conditions (International Association on
Water Quality (IAWQ), 1998). The present study aimed to use a re-
spirometric assay to determine the performance of synthetic fibres as
biological support for biofilters in recirculating freshwater systems for
fish.
2. Material and methods
2.1. Description of the recirculating experimental system
The experiment was developed in the AQUOS Laboratory, UFSC, SC,
Brazil (27°35′49′' S, 48°32′58′' W). It involved the use of a circular
plastic 2,400-L tank that had 1920 L of usable volume. The system had a
centrifuge pump (Eletroplas® model, ICS 50 BV) that was responsible
for the transport of water from an external 200-L stocking tank to the
internal 300-L reservoir used for water distribution. The system was
installed 2 m above the fish tanks, and it had a mean water flow rate of
1900 L/h.
Treatment of the water for mechanical and biological filtration re-
quired the use of a settler and four 200-L plastic tanks. The settler re-
moved the larger particles and had a capacity of 120 L and a conical
bottom. The first plastic tank was able to remove the physical solid
sediment. The second tank contained previously cleaned and disin-
fected oyster shells to maintain the alkalinity of the system (Summerfelt
et al., 2015). The third and fourth tanks were used for biological fil-
tration, and they contained 200 units of synthetic fibre
Fig. 1. Apparatus used to measure respirometry of the synthetic fibre used as biological support.
57
Aquacultural Engineering 82 (2018) 56–62
(225 mm x 110 mm x 10 mm) as media for biological support.
2.2. Technical characteristics of biological support
The biological support used in the assay was composed of synthetic
fibres and abrasive material, a waterproof polyester resin that disable
the contact of water with minerals. It was designed in a reticulated
manner like a web constituted of resistant polyethylene.
2.2.1. Colonisation of synthetic fibre
For the nitrifying microorganism colonisation assay, the re-
circulating system was filled with 3320 L of previously chlorinated
(5 ppm of sodium hypochloride) potable water, and it was de-
chlorinated with sodium thiosulphate (1 ppm). Water quality was
monitored daily with a multiparameter meter (Hanna Instruments
model HI 9828) to measure temperature, dissolved oxygen and pH and
with a colorimetric kit (Acqua Supre) to measure alkalinity, ammonia
and nitrite.
During the assay, there was no water exchange, but there was water
replacement (maximum of 1% weekly) for any loss that occurred due to
evaporation. To generate a degradation substrate for nitrifying micro-
organisms, a broth containing 200 g of a commercial food for fish,
KOWALSKI® (crude protein, 350 g/kg), was used as the initial source of
nitrogen. This amount was calculated as adequate to be offered to 120
fish (estimated biomass of 3.8 kg at the beginning of the assay). The diet
was placed in a 400-mL beaker of water until complete the pellets were
destabilised, which allowed the soluble nitrogenated compounds to
leach into the liquid fraction. After that, 100 mL of the broth, which
contained soluble nutrients, was added to the system to increase the
ammonia concentration. In total, four pulses of 100 mL of diet broth
were added over 10 days in an effort to colonise the synthetic fibre.
The first inoculum of nitrifying bacteria was added from a com-
mercial product (NITE-OUT II®) that contained Nitrosomonas,
Nitrobacter and Nitrospira. Its basic composition, as provided by the
manufacturer, was 40% nitrifying bacteria, 58% water, 1% sodium
bicarbonate and less than 1% preservative. A total of 250 mL of the
inoculum were added to tanks three and four. For the addition of the
commercial product, the recirculating water system was stopped for 2 h
so that the nitrifying bacteria could adhere on the synthetic fibres in the
tanks. The duration of the colonisation assay was 10 days.
2.2.2. Respirometry assay
Seven days after use of the system began, when the total ammonia
level and nitrite remained reduced, we began the respirometry assay to
verify the presence of nitrifying chemoautotrophic microorganisms.
The setup of the equipment was adapted from Spanjers et al. (1996),
M.S. Owatari et al.
and it included a 350-ml Erlenmeyer flask employed as a bioreactor
(Fig. 1).
Fig. 1
For this assay, one synthetic fibre was removed from the tank, and a
smear was made to remove the biomass of microorganisms that had
adhered to the surface. The media was washed with the aid of a specific
culture medium for nitrifying bacteria, as described by Campos et al.
(1999). The final concentration was adjusted to 1 g of total suspended
solids (SST)/L (according to the culture composition reported by
Campos et al. (1999)). The sample was washed, and 300 mL of the 1 g
SS/L suspension was transferred to the respirometry equipment. After
the addition of the bacterial biomass, the system was aerated with an
air compressor, and the pH was adjusted to 7.8–8.0 with a 2.5% (w/v)
sodium hydroxide solution (NaOH) and a 2.5% (v/v) hydrochloric acid
(HCl) solution. The temperature was maintained at 35 °C and a mag-
netic agitator was used at 300 rpm during the respirometric assay.
After aeration was interrupted, the determination of endogenous
respiration began. The dissolved oxygen was measured at 30 s intervals
using an oxymeter (YSI 55, Pro 20). The percent saturation was con-
trolled so that is remained above 30% in order to not compromise the
bacterial activity. After that, aeration was re-established (International
Association on Water Quality (IAWQ), 1998).
Respiration of the bacterial biomass was determined by using an
ammonium chloride solution (NH4Cl). First, a pulse of 1 g/L of am-
monium chloride was made to the system. The aeration was inter-
rupted, and the oxygen was read at 30 s intervals. A total of four pulses
of ammonium chloride were used: concentration [C1]35.369 mg NH3/
L; [C2]51.628 mg NH3/L; [C3]79.494 mg NH3/L; and [C4]215.225 mg
NH3/L.
Ammonia determination followed the Nessler colorimetric method
described by Vogel (1981), whereby the Nessler reagent was added to
the sample in a proportion of 1:50. After 10 min, the sample was read at
525 nm absorbance using a spectrophotometer (HAC DR 5000). With
the absorbance value, the ammonia concentration was verified using a
standard curve that was obtained from the ammonium chloride solu-
tion. The respiratory velocity (that is, the oxygen consumption) caused
by the substrate degradation was expressed by the following equation:
r0 = QO2 X (1)
where,
r0 = respiration rate or oxygen consumption;
QO2 = specific breathing rate (mg O2/g of cells per hour); and
X = cell concentration (g of cells per L).
Thus, the mass balance for oxygen in this system was:
dC
= Kla (Cs−C )−QO2 X
dt (2)
Where,
Cs = concentration of O2 at saturation;
C = concentration in the liquid; and
the term Kla (CS-C) relates to the oxygen transfer.
When the transfer was cancelled by eliminating the aeration, we
obtained:
dC
= − QO2 X
dt (3)
Which resulted in the following integrated equation:
C = C0−QO2 X * t (4)
Therefore, the linear model between the dissolved oxygen concentra-
tion (C) and time (t) was determined.
With the data obtained, a graphical model of
Michaelis–Menten–Monod kinetics (Monod, 1949) was constructed.
The initial velocity of the reaction was measured using the substrate
concentrations as the scale (nominated as [S]); the velocity of the re-
action (V) increased as [S] increased. Furthermore, as [S] increased, the
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Aquacultural Engineering 82 (2018) 56–62
enzyme saturation and the velocity reached a maximum value (Vmax),
according to the following equation:
QO2 max . [N −NH3]
QO2 =
[N −NH3] + Ks (5)
where,
Ks = the affinity for the substrate mg[N-NH3]/L.
It is important to emphasise that the oxygen consumption that oc-
curred due to endogenous respiration and degradation of the biomass
substrate was necessary to differentiate the two drops. The QO2X found
was subtracted from the value of the endogenous drop.
2.2.3. Scanning electron microscopy
In order to verify the synthetic fibre structure used as a biological
support and the bacterial colonisation, samples of the fibre were col-
lected from the third and fourth tanks (biological filters) at the same
time that the respirometry assay was performed. Samples were fixed in
2.5% glutaraldehyde solution, 2.0% sucrose and buffered with 0.1 M
cacodylate (pH 7.2) (Schmidt et al., 2010). After that, they were de-
hydrated in a graded ethanolic series, submitted to critical-point drying
in a EM-CPD-030 (Leica, Heidelberg, Germany) equipment, mounted in
specimen stubs and metalized with gold (Metalizador Blatec, CED 030).
Photomicrographs were obtained using a Jeol 6390 L V microscope (a
20 kV; JEOL Ltd., Tokyo, Japan).
2.3. Fish harvesting in the experimental tanks
At the end of the colonisation assay, an experimental unit was filled
with 120 Nile tilapia (Oreochromis niloticus). The fish samples were
acquired from a fish farm located in the municipality of Presidente
Getúlio, SC, Brazil. The fish in the system had a mean weight of
32.11 ± 7.60 g, which represented a total biomass of 3.8 kg and a
stocking density of 1.97 kg/m³. During the 60 days of the experiment,
they were fed four times per day with a commercial diet (Kowalski®)
that contained 35% crude protein at 5% of the biomass.
After analysing the water quality parameters and zootechnical in-
dices obtained from the tilapia, we designed another assay with the goal
of increasing the biomass in order to compare the biological filter ef-
ficacy. After the tilapia assay, the system was given a sanitary break
before the new assay was set with 480 jundiá (Rhamdia quelen). The fish
for this second assay were acquired from a commercial fish farm located
in Lages, SC, Brazil. The fish in the system had a mean weight of
11.34 ± 2.4 g, which represented a total biomass of 5.4 kg and a
stocking density of 2.81 kg/m³. During the 60 days of the experiment,
they were fed four times per day with a commercial diet (NICOLUZZI®)
that contained 42% crude protein at 5% of the biomass.
All animal procedures were approved by the Ethic Committee on
Animal Use – CEUA/UFSC nº PP00928 and 7119220216.
2.4. Statistical analysis
For the respirometry analyzes, a simple linear regression between
time and oxygen consumption was performed. The data obtained in the
experiment were analyzed with statistica® 13.0 software.
3. Results and discussion
Maturation of the biological filter was observed in the first days
after inoculation of the microorganisms. The analysis of total ammonia,
toxic ammonia and nitrite showed decreased values, which confirmed
the nitrification process. The maximum and minimum values of total
ammonia nitrogen (TAN) during this period were 3.6 mg/L and 0.6 mg/
L, respectively. Changes to the TAN values were observed due to the
inclusion of a nitrogen source in the system as a result of the addition of
the diet broth diluted in the water. Ten days after colonisation of the
synthetic fibres, a reduction was observed in the total ammonia of
M.S. Owatari et al.
Fig. 2. Temporal variation of total ammonia concentration in the first days of the experiment after colonizing the synthetic fibre with diet as biological filter. Arrows
indicate the addition of fluid diet when observing decrease of the total ammonia nitrogen (TAN).
between 35 mg/L and 1.2 mg/L (Fig. 2).
Kuhn et al. (2010) used a commercial product (PondProtect-L®) for
marine shrimp that was composed of nitrifying bacteria (15 ppm) as the
initial bacterial inoculum of the biofilter with Kaldnes media. They
reported nitrifying activity two days after the product was added to the
system. They also reported that total ammonia did not exceed the
concentration of 1.0 mg/L during the 11 days of the experiment.
Cycles of aquaculture production normally depend on auto-
chthonous nitrifying bacterial colonisation in the biofilters, but con-
siderable time is needed for the establishment of healthy and viable
nitrifying bacteria (Kuhn et al., 2010). The use of a commercial product
to start the process has shown to be an efficient protocol for media
colonisation in the present study.
Fig. 2
3.1. Respirometry and specific oxygen consumption
In the respirometry assay, the oxygen consumption over time was
verified after the aeration was interrupted (Figs. 3 and 4). Simple
methods for the use of respirometry have been previously reported;
they emphasise the evaluation of a respirogram (Spanjers et al., 1999)
and using less sophisticated equipment on a small scale (Gernaey et al.,
1997; Spanjers et al., 1996; Xu and Hasselblad, 1996).
Figs. 3 and 4
In observing our results, the QO2 and an increase in the respiration
activity of the microorganisms was found to be directly related to the
Fig. 3. Relationship between oxygen consumption along the time in the endogen respiration of the microorganisms removed from the synthetic fibre. The values
were registered at 30 s interval during the respirometry assay.
59
Aquacultural Engineering 82 (2018) 56–62
addition of the substrate (Fig. 5). This value increased as the ammonia
concentration increased in the environment, and through the re-
spiratory quantification of the biomass, the capacity to enhance the
biodegradability of organic substances could be determined (Urfer and
Huck, 2001).
Fig. 5
The kinetics of oxygen consumption is an important factor to pro-
mote more confidence to this test by showing the metabolic activity of
the microbiota present in the environment. A study by Spanjers et al.
(1996) showed that linearity is normally observed in this process. In
other words, there is a logical sequence between the substrate and
oxygen demand, as supported in the present assay. The results de-
monstrate a linear relationship between the amount of substrate added
to the system and the oxygen demand.
Costa et al. (2007) described respirometry as a useful tool to eval-
uate the metabolic capacity of microorganisms because it is practical
and reliable for kinetics studies. Gernaey et al. (2001) used re-
spirometry to compare the oxygen consumption in activated sludge
with different concentrations of organic material, and they found a
constant linear relation between inclusion of the substrate and oxygen
consumption, which corroborated the present results. It would be fea-
sible to apply respirometry tests to evaluate the characteristics of re-
sidual water from aquaculture systems on the basis of the microbial
activity.
Colonisation of the synthetic fibres was also verified by scanning
electron microscopy, making it possible to observe different
M.S. Owatari et al.
Fig. 4. Relationship between oxygen consumption along the times by the microorganisms removed from the synthetic fibre with the presence of substrate ammonium
chloride (NH4Cl). The values were registered at 30 s interval during the respirometry assay after included the ammonium chloride ([c]1 = 35.369 mg.NH3 L−1; [c]
2 = 51.628 mg.NH3 L−1; [c]3 = 79.494 mg.NH3 L−1; [c]4 = 215.225 mg.NH3 L−1).
Fig. 5. Oxygen consumption in relation to time with an increase in the am-
monium chloride supply.
morphotypes of bacteria 10 days after media colonisation. Apart from
no measurements of the specific surface area, ramified structures could
be found, which confirmed the biological support (Fig. 6).
Morphological characteristics and the action of the bacterial co-
lonies in the nitrifying process into biological reactors under laboratory
conditions was described by Schramm et al. (1998) during a study
identifying the nitrifying bacteria as Nitrosospira and Nitrospira spp. In
the present study, although there was no identification of the micro-
organisms present in the biological surface, our results from the re-
spirometry assay involving pulses of inorganic nitrogen without the
presence of organic carbon made it possible to infer that the che-
moautotrophic process did occur. During that 120 days of the experi-
mental period, the efficiency of the biological filter with synthetic fibres
was tested, and results showed that the tilapia and jundiá reached sa-
tisfactory weight gain and survival under adequate water quality.
3.2. Assay with fish in the system
For O. niloticus and R. quelen, in the present study was verified a
60
Aquacultural Engineering 82 (2018) 56–62
final biomass of 12.6 g and 21.5 g, with specific growth of 1.98 g and
2.29 g, respectively (Table 1). Testing several plastic materials as bio-
logical support for tilapia (120 g), the fish were fed three times per day
with diet containing 36% crude protein in a recirculating system, Al-
Hafedh et al. (2003) observed a mean daily growth of 2 g, similar to
that found in the present study. In the studies of Canton et al. (2007),
fish were fed four times per day and reached a mean weight of
41.10 ± 4.56 g, with a specific growth rate of 1.56% per day and
survival of 80.12%. In contrast, our study showed higher values for
jundiá due to the controlled conditions of the study. In fact, this ex-
plains the best growth performance found in this study when compared
to other studies.
After 70 days of the jundiá assay, the parameters of the filtration
system did not show oscillations as occurred at the beginning of the
assay. This was due to the fact that the bioreactors were colonised with
nitrifying bacteria from the initial inoculum into the biofilter
(Blancheton et al., 2013).
Al-Hafedh et al. (2003) described the performance of different
plastic materials used as biological support (for example, plastic roll,
PVC pipe and a cleaning sponge) in biofilters of recirculating systems.
The Al-Hafedh et al. (2003) study used Nile tilapia at an initial stocking
density of 20 kg/m3, and at a final density of 39 kg/m³, and the fol-
lowing conditions were reported: water temperature = 24.36 ± 0.4 °C;
dissolved oxygen (DO) = 6.12 ± 0.23 mg/L; pH = 7.77 ± 0.05; total
ammonia nitrogen (TAN) = 0.92 ± 0.05 mg/L; and nitrite (N-
NO2) = 0.22 ± 0.04 mg/L. On the other hand, Guerdat et al. (2010)
evaluated three biological filter models (including, moving bed bior-
eactor, floating bead filter, and fluidised sand filter) available com-
mercially for recirculating systems in aquacultures. The Guerdat et al.
(2010) study reported the use of 5000 Nile tilapias at a stocking density
of 18.58 kg/m³ and the following water-quality parameters: tempera-
ture = 28.9 ± 2.3 °C, DO = 5.8 ± 0.8 mg/L, pH = 7.16 ± 0.14,
TAN = 0.69 ± 0.3 mg/L and N-NO2 = 2.1 ± 1.5 mg/L. In the present
study water-quality parameters were similar in both assays tested
M.S. Owatari et al. Aquacultural Engineering 82 (2018) 56–62

Fig. 6. Scanning Electron Microscopy (SEM) of the structure of biological media synthetic fibre-like. Bacterial morphotypes adhered to the synthetic fibre (A) and 10
days after colonization (B).

Table 1 tilapia growing under sterile conditions and the primary maturation of
Zootechnical parameters (means+standard deviation) of Oreochromis niloticus the biofilter before the experimental assay. In the assay with the jundiá,
and Rhamdia quelen in the recirculating aquaculture system. the recirculating system was mature, and the water-quality parameters
Zootechnical parameters Oreochromis niloticus Rhamdia quelen were kept more homogenous with no oscillations.

Number of fish (unit) 120 480 4. Conclusion

Initial total biomass (kg) 3.8 5.4
Initial weight (g) 32.11 ± 7.6 11.34 ± 2.40
Total final biomass (kg) 12.6 21.5 The fibre evaluated in the present study was efficient as biological
Final weight (g) 105.53 ± 34.25 44.79 ± 9.38 support in the recirculating aquaculture system for tropical fish. It
Weight gain (g) 73.42 ± 26.65 33.45 ± 6.98 supported a stocking density of 6.56 kg of O. niloticus/m³ and 11.19 kg
Specific growth rate (% day) 1.98 2.29 of R. quelen/m³, and it contributed to the maintenance of good water
quality.
Table 2
Acknowledgements
Mean values and standard deviation of the water quality during 60 days of fish
maintenance in the recirculating aquaculture system (RAS). DO: dissolved
oxygen, T: temperature, TAN: total ammonia nitrogen. The authors thank National Council of Scientific and Technological
Development (CNPq) for research grant and financial support to J.L.P.
Parameters Oreochromis niloticus Rhamdia quelen
Mouriño (CNPq 308292/2014-6) and M.L. Martins (CNPq 446072/
DO (mg L )−1
6.97 ± 1.25 6.70 ± 0.41 2014-1, 305869/2014-0); Chemical Engineering Department (UFSC)
pH 6.78 ± 0.46 7.27 ± 0.33 for respirometry assay; Central Laboratory of Electronic Microscopy
T (°C) 24.41 ± 1.25 28.81 ± 0.83 (LCME, UFSC) for Scanning Electron Microscopy and Alevinos do Sabiá
TAN (mg L−1) 1.45 ± 1.21 0.17 ± 0.15
fish farming for fish donation.
Tóxic ammonia (mg L−1) 0.004 ± 0.003 0.003 ± 0.003
Nitrite (mg L−1) 1.25 ± 0.98 0.56 ± 0.61
Alkalinity (mg L−1) 38.90 ± 21.19 19.35 ± 4.47 References

(Table 2), with an initial biomass of 1.97 kg/m³ (tilapia) and 2.81 kg/
m³ (jundiá), and a final biomass of 6.56 kg/m³ and 11.19 kg/m³, re-
spectively. We suggest more tests with these media using higher
stocking densities.
Colt et al. (2006) have related great variation of studies that sta-
tistically define a stabilised biofilter. On the other hand, some authors
argue that a stable biofilter must be capable of keeping the TAN and N-
NO2 below 0.7 mg/L, with an ammonia load of 8.64 g, during seven
consecutive days (Sandu et al., 2002), and the total ammonia–nitrogen
load removed must be equal to the load influx in the system (Zhu e
Chen, 2001). It was possible to verify the standard stability of the
biofilter; this was accomplished mainly during the second assay with
jundia, during which the ammonia load was 1.6 g and total ammonia
nitrogen and nitrite was kept below 0.5 mg/L during almost the entire
experimental period.
Total ammonia presented characteristic kinetic patterns in each
period; however, clear variations were observed at the beginning of the
assay with tilapia, whereby decreased TAN occurred after the peaks
registered. It is possible that this observation occurred as a result of the
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