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Aquacultural Engineering 58 (2014) 2028

Contents lists available at ScienceDirect

Aquacultural Engineering
journal homepage: www.elsevier.com/locate/aqua-online

Culture of yellowtail kingsh (Seriola lalandi) in a marine recirculating

aquaculture system (RAS) with articial seawater
J. Orellana a, , U. Waller b , B. Wecker c
a

Escuela de Ciencias del Mar, Ponticia Universidad Catlica de Valparaso, Av. Altamirano 1480, Valparaso, Chile
Hochschule fr Technik und Wirtschaft des Saarlandes, Goebenstr. 40, 66117 Saarbrcken, Germany
c
neomar GmbH, Am Osterberg 22, 31311 Uetze-Eltze, Germany
b

a r t i c l e

i n f o

Article history:
Received 18 June 2013
Accepted 17 September 2013
Keywords:
Seriola lalandi
RAS
Growth
Nitrication
Denitrication

a b s t r a c t
The development of new species is a high priority for the diversication of the Chilean aquaculture
sector. The yellowtail kingsh (Seriola lalandi) is a promising candidate for commercial production in
recirculating aquaculture systems (RAS). This paper presents data on the culture of yellowtail kingsh
in a marine RAS working for 488 days using articial sea water. Growth performance, feed conversion,
feeding rate, condition factor and mortality were determined for sh having an average initial weight
(S.D.) of 0.7 0.2 g up to a nal average weight of 2006 339.0 g. The RAS conguration (drum lter,
protein skimmer with ozone, biological nitrication and denitrication, carbon dioxide removal and
oxygenation) showed performance stability under the conditions assayed (low water renewal rate). Total
ammonia nitrogen and nitrite-nitrogen concentration averaged 0.74 0.42 mg/L and 0.21 0.24 mg/L
respectively. After installation, the denitrication reactor kept nitrate-nitrogen concentrations below
40 mg/L. Nitrate-nitrogen was totally reduced at oxidation reduction potential values between 150
and 250 mV. Water temperature averaged 22.6 1.4 C and oxygen was maintained close to saturation
levels. Carbon dioxide concentration was in average 8.3 2.47 mg/L and pH 7.5 0.1. Water renewal rate
was 0.45% of the total system volume per day. The system proved the capability to maintain optimal
water quality and secured animal welfare.
2013 Elsevier B.V. All rights reserved.

1. Introduction
The development of new nsh species is a high priority for the
diversication of the aquaculture sector in Chile, in order to expand
the production to high-value resources and different geographical zones. Around 95% of the Chilean aquaculture sh production
is salmon and the culture activities are principally focused in one
region. As part of the Chilean aquaculture diversication Program
(Programa de Diversicacin de la Acuicultura Chilena, PDACH)
the yellowtail kingsh (Seriola lalandi) has been investigated for
culture and ngerlings are commercially produced in recirculating aquaculture systems (RAS) from closed life cycle spawning
since 2009. Most of the juvenile production has being destined to
growth trials abroad, mainly in RAS. Such systems offer the necessary bio-security for the culture of a non-native species, water
quality control as well as waste management. Biosecure RAS also
avoids disease outbreaks and parasites because of the lack of intermediate hosts.

Modern closed RAS can operate with articial seawater and less
than 1% of water renewal per day. These high-tech systems allow
the land-based cultivation of exotic species of high commercial
interest, close to the consumer, and with zero discharge of nutrients
and organic matter into natural ecosystems when combining with
multi-trophic integrated aquaculture (IMTA). Additionally, product
traceability is possible. This type of technology is environmentally sound and contributes to the sustainability of aquatic food
production.
The purpose of this study was to provide data on the culture
of yellowtail kingsh in a marine RAS. There appear to be no published data for growth performance of yellowtail kingsh in a closed
RAS operating with articial sea water, and less than 1% of water
renewal per day. This data is of importance for optimizing both,
sh and system performance. In view of the available literature,
this study seems to be the very rst investigating the feasibility of
RAS culture of yellowtail kingsh.
2. Materials and methods
2.1. Fish

Corresponding author. Tel.: +56 32 2274239.

E-mail addresses: jaime.orellana@ucv.cl (J. Orellana),
uwe.waller@htw-saarland.de (U. Waller), bert.wecker@neomar.de (B. Wecker).
0144-8609/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.aquaeng.2013.09.004

S. lalandi ngerlings (n = 2000) were transported from the company Acuicola del Norte S.A., Caldera, Chile to neomar GmbH,

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J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028
Table 1
Tap water quality parameters, determined according to the analytical methods
established by the German drinking water regulation (Trinkwasserverordnung TVO
of 2001) in accordance with the European Union Council Directive 98/83/EC of 1998.
Maximum permissible values (in cursive) are presented.
Parameter
Conductivity

pH

Turbidity

(S/cm)

(FNU)

2500
557

6.59.5
7.69

1.0
0.07

Cl
250
57.8
Pb
0.01
<0.001

S
240
148
Cd
0.005
<0.001

Fe
0.2
0.017
Cr
0.05
<0.001

Mn
0.05
<0.001
CN
0.05
<0.005

13.19

NH4
0.5
<0.04
F
1.5
0.16

2.36

NO2
0.5
<0.03
Cu
2
<0.01

NO3
50
2.0
Ni
0.02
0.0063

Ar
0.01
<0.001
Hg
0.001
<0.0001

Formazine nephelometric units.

Table 2
Element composition in percentage by weight of the lab-made sea salt.
Ingredienta

Quantity (%)

Chloride
Natrium
Sulfate
Magnesium
Potassium
Bicarbonate
Carbonate

55.69
30.31
7.12
3.15
1.13
0.57
0.05

Element (mg/L)
NO3
<14

NH4
<14

B
368

Cu
<64

P
<12

K
469

Ca
485

Mg
1361

S
946

(g/L)

(mmol/L)

Element (mg/L)
Na
200
31
Al
0.2
<0.02

Table 3
Concentration of major and trace water-soluble elements, pH, and electrical conductivity (EC) in the mixed articial sea water.

Na
9920

Cl
18,930

Total hardness
( dH)

21

a
Trace elements (Cu, Fe, Mo, Mn, Zn and B) are included within major salt components.

Uetze-Eltze, Germany, at a size of approximately 0.5 g. After 50 h

of transport, sh were received and transferred into the recirculating system at 21.3 C, 32 S, pH of 8.05, redox potential of 243 mV,
95% oxygen saturation, and a photoperiod of 12:12 h light:dark. The
ngerlings were fed daily with commercial pellets. The rst 35 days
pellets containing 52% protein and 20% lipids (Nutra, Skretting, The
Netherlands) were fed at a ratio between 9.0 and 8.1%BW/d, followed by pellets containing 55% protein and 16% lipids (Marico
Mistral, Coppens, The Netherlands), at a feeding ratio between 1.9
and 6.8%BW/d until day 166. Afterwards, pellets containing 50%
protein and 15% lipids (Europe 15, Skretting, The Netherlands) were
fed at 0.61.5%BW/d. Changes in feed type and size are detailed in
Table 4.
2.2. System design
The study was carried out in a RAS working with articial
seawater and less than 1% of water replenishing rate per day.
Seawater was prepared by mixing tap water with a lab-made
salt mixture. The results of the tap water analysis are detailed
in Table 1. The salt composition (Table 2) contains mainly the
seawater elements given by Spotte (1979). The concentration of
major and trace elements in the mixed articial seawater are
presented in Table 3. The RAS (Fig. 1) comprised of a drum lter with a 60 m screen panel (Hydrotech HDF 501-1P, Velinge,
Sweden) and a protein skimmer (Sander Helgoland 500, UetzeEltze, Germany) for the removal of large and ne particulate waste
respectively. The protein skimmer was operated with ozone to
enhance the removal of ne solids, reduce bacteria load and oxidize
nitrite and colour, as reported by Bullock et al. (1997), Summerfelt
and Hochheimer (1997), Summerfelt et al. (1997), Krumins et al.
(2001a), Summerfelt (2003), and Tango and Gagnon (2003). Water
temperature was maintained through nine immersion heaters

Fe
<112

Mn
<55

Mo
<96

Zn
<65

pH

EC (mS/cm)

8.3

46.5

(Shego 300 W, Offenbach, Germany) and a cooling unit (AquaMedic

Titan 2000, Bissendorf, Germany). A moving bed bioreactor (MBBR,
AnoxKaldnes, Lund, Sweden) was used for ammonia removal using
K3 media. The sh tank (7.3 m3 ) was supplied with oxygen gas
through a ceramic diffuser. Carbon dioxide was removed through
water cascades at the outlet of the biolter. Calcium hydroxide
(lime) slurry was added for pH stabilization, controlled by the pH
reading in the sump. For the removal of nitrate a denitrication
unit was installed 40 days after the start of the experimental run.
The anaerobic biolter was supplied with acetic acid as hydrogen
proton donor and carbon source.
2.3. Measurements
During the trial, dissolved oxygen and temperature were
measured with a multimetre (Hach Lange sc1000), pH and
oxidationreduction potential (ORP) were measured with probes
(Brkert 8205 and 8206 respectively), carbon dioxide with the
OxyGuard CO2 metre, salinity with a refractometer, and hardness with a test kit (Tetra). Total ammonia nitrogen (TAN, Nessler
method), nitrite-nitrogen (NO2 -N, dizotization method), nitratenitrogen (NO3 -N, cadmium reduction method), phosphate (PO4 ,
amino acid method) and total residual oxidants concentration (TRO,
DPD method) were measured using a spectrophotometer (Hach
Lange DR2000). All parameters were determined daily at approximately 8:00 h, with the exception of TRO, that was measured at
irregular intervals. Water samples were taken from the sump and
from the sh tank when TRO was determined. After the installation
of the anaerobic biolter, an additional water samples was taken
from this unit to determine the concentration of NO2 -N and NO3 -N
to evaluate denitrication rate.
Growth performance was assessed as specic growth rate (SGR)
expressed as percentage per day (%/d), described by the formula:
SGR = (ln(Wf ) ln(Wi ))

100
t

(1)

where Wf , nal weight (g); Wi , initial weight (g) and t, time between
measurements (d).
Additionally, fork length was measured with every weight
sample to determine condition factor (K), based on the review presented by Froese (2006) and using the formula:
K=

W
100
L3

(2)

where W, individual weight (g); L, fork length (cm)

Feed conversion ratio (FCR) was calculated for every sample
period as the relation between feeding rate and SGR. Fish (randomly
selected from the tank, N = 50 in average) were anaesthetized using
eugenol oil (Oleum caryophylli) at a dose between 0.3 and 1.8 mL per
100 L seawater. After sampling, sh were returned to the tank.
3. Results
After arrival and opening the transport bags, water and sh
were distributed into a small 600 L tank prior to introduction into

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J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028

Fig. 1. System set-up of the experimental RAS used in this study. Water, air and oxygen ows are indicated with arrows. Water parameters such as oxidationreduction
potential (ORP), temperature (TEMP), salinity (S), hardness ( dH), carbon dioxide (CO2 ), and pH are detailed in the respective measuring points.

the RAS. The transport water had a pH of 7.5 and a temperature

of 21.2 C. Oxygen saturation amounted 101%, salinity 32, hardness 15 dH and CO2 11.1 mg/L. TAN and NO2 -N was 3.7 mg/L and
0.09 mg/L respectively. Water quality was slowly adjusted by mixing transport water and RAS water.

Daily TAN and NO2 -N concentrations in the experimental RAS

are presented in Fig. 2A and B respectively. TAN concentration
averaged 0.74 0.42 mg/L (min 0.17 mg/L and max 2.75 mg/L)
and NO2 -N concentration averaged 0.21 0.20 mg/L (min
0.01 mg/L and max 2.40 mg/L) over 488 days. High TAN peaks were

TAN

Nitrite-N
3.0

A
Concentration (mg/L)

Concentration (mg/L)

3.0

2.0

1.0

0.0

2.0

1.0

0.0
0

100

200

300

400

500

100

200

Days
Nitrate-N

400

500

Phosphate

160

60

D
Concentration (mg/L)

140
Concentration (mg/L)

300

Days

120
100
80
60
40

40

20

20
0

0
0

100

200

300

Days

400

500

100

200

300

400

500

Days

Fig. 2. Total ammonia nitrogen (TAN) (A), nitrite-nitrogen (B), nitrate-nitrogen (C), and phosphate (D) concentration in the RAS over 488 days of experimentation.

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J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028
114

106

Temperature (C)

O2 saturation (%)

30

110

23

102
98
94
90

25

20

86
82

15

100

200

300

400

500

100

200

Days

300

400

500

Days

Fig. 3. Oxygen (O2 ) concentration (A) and water temperature (B) in the RAS over 488 days of experimentation.
8.2

20

7.8

pH

18

CO2 concentration (mg/L)

8.0

7.6
7.4
7.2
7.0
6.8

16
14
12
10
8
6
4
2
0

100

200

300

400

500

Days

100

200

300

400

500

Days

Fig. 4. pH (A) and carbon dioxide (CO2 ) (B) concentration in the RAS over 488 days of experimentation.

occasionally observed, related to changes in the feed type and size,

and systems performance. NO2 -N peaks (above 1.0 mg/L) were
observed between day 28 and day 80 of the experiment. After this
period NO2 -N concentration decreased and values mainly below
0.5 mg/L were observed.
An accumulation of NO3 -N and PO4 in the system (Fig. 2C and
D) was observed during the rst 90 days of the experiment. Maximum NO3 -N concentration reached 154.5 mg/L on day 86. Fish
feed intake was negatively inuenced by increasing NO3 -N concentration and therefore, around 50% of the system water was
exchanged. The anaerobic biolter was brought into operation on
day 40. NO3 -N removal rates were observed from day 100 on. Later
on, NO3 -N concentrations rose to values above 100 mg/L around
day 360 and again around day 380. This corresponds to events
related to a substantial drop in the pH to values around 4.0 inside
the denitrication reactor in both cases.
Fig. 3A shows the daily dissolved oxygen concentration for the
study period indicating uctuations between days. The sh tank
was continuously supplied with oxygen gas through a ceramic diffuser, controlled by a proportional valve and a threshold of 98%
saturation. Average oxygen amounted to 97.6 3.2% saturation
(min 83.6% and max 113%). After day 363 and until the end of the
experiment, the oxygen control unit was re-set to a threshold of
100% saturation, which explains the change in the range of the
daily uctuations in the recorded data. Temperature uctuations
(Fig. 3B) differed between summer (between days 180 and 310) and
winter although room temperature was controlled during winter
by heating (20 C) but somewhat dependant on external weather
conditions. Water temperature averaged 22.6 1.4 C (min 15.1 C
and max 28.3 C). Extreme high values reects the under designed
cooling unit during summer time, approximately between days 180
and 310. The switch-on of the immersion heaters during winter
time, at the beginning and at the end of the experiment, resulted
in proper temperature control.
Daily pH values are presented in Fig. 4A. Mean pH was 7.5 0.1,
however, the data have to be assessed on the time scale as the effect
of pH on other water quality parameters is crucial. Maximum and

minimum values reached up to 8.1 and 6.9 respectively. Because of

the pH controlled addition of lime (calcium hydroxide suspension),
pH values were maintained between 7.4 and 7.8 at all times with
little daily uctuations. Towards the end of the study period, single
pH values above 7.8 were observed.
Fluctuations in the carbon dioxide concentration (Fig. 4B)
were observed between daily measurements. An increment was
recorded from day 1 and until day 80 approximately. Average CO2
concentration was 8.3 2.47 mg/L. Highest concentration measured was 17.1 mg/L on day 232. Degassing was affected by water
temperature, depicting the uctuations in the CO2 concentration
during summer time (between days 180 and 310) in relation to the
depletion during winter, with the highest stocking density towards
the end of the experiment.
Daily ORP in the protein skimmer (Fig. 5A) and in the pump sump
(Fig. 5B) averaged 332 62 mV and 202 30 mV respectively. The
dosage of ozone into the protein skimmer was controlled by an ORP
threshold set at 300 mV during the rst 80 days, then at 320 mV
until day 175, at 360 mV until day 182, and at 400 mV until the
end of the experiment. High variations in the daily ORP readings
were observed for the protein skimmer (Fig. 5A). In contrast, the
pump sump showed a more stable ORP values during the whole
experiment, despite some very low single readings below 150 mV
(Fig. 5B).
The relative NO3 -N removal in relation to the ORP value in
the denitrication reactor is depicted in Fig. 6. The daily NO3 -N
removal efciency in relation to the measured ORP value shows
that, the optimal window for a complete NO3 -N reduction to
gaseous nitrogen begins at ORP values below 150 mV. During
operation ORP may fall to values around 250 mV. ORP values
above 100 mV resulted in an incomplete denitrication process,
where only 2030% of the NO3 -N was removed.
Fig. 7A shows the increment in mean total weight of the sh
over time during the trial period. Fish were reared from an average weight of 0.74 0.18 g to 2006 339.03 g in 488 days. At the
end of the experimental period, the RAS had a stock of 374 sh and
a density of 103.5 kg/m3 (Table 4). The relationship between mean

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J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028

ORP sump
350

300
250

ORP (mV)

ORP (mV)

ORP skimmer
600
550
500
450
400
350
300
250
200
150
100
50
0

200
150
100
50
0

100

300

200

400

100

500

Days

200

300

400

500

Days

Fig. 5. Oxidationreduction potential (ORP) inside the protein skimmer and in the pump sump of the experimental RAS, over 488 days of experimentation.

Relative NO3-N removal (%)

120%

Water replenishing rate amounted to 0.45% of the total system

volume per day, on average. Daily water loss mainly corresponded
to the formation and discharge of foam through the protein skimmer, and through the water content in the sludge removed by the
micro screen drum lter.

100%
80%
60%

4. Discussion

40%
20%
0%
-400

-300

-200

-100
ORP (mV)

100

200

Fig. 6. Relative nitrate-nitrogen (NO3 -N) removal (reduction to N2 ) in relation to the

oxidationreduction potential (ORP) measured in the outow of the denitrication
reactor.

fork length and mean weight is graphically presented in Fig. 7B. The
isometric growth of S. lalandi is explained by the value of the parameter b (2.99) and the coefcient of determination of the correlation
(R2 = 0.99).
Table 4 summarizes the growth performance of S. lalandi. The
initial value of the condition factor (K) shows a good recovering
after transportation stress before stocking and a good adjustment
to gradually appropriate feeding levels. Specic growth rate (SGR)
declined with weight increase. The feeding of the sh was carried
under commercial conditions i.e., maximum feeding amount for
optimal growth. Fish were fed normally through automatic feeders.
Feeding rate (%BW) decreased over time. FCR was affected by mortalities when sh died after weight determinations or harvesting.
Some events related to high CO2 concentration and high temperatures during summer time had an inuence on feed intake. The
highest observed FCR of 6.7 remains unexplained.
2500

2500

2250
1750
1500
1250
1000
750
500
250

W = 0.0225 L 2.9908
R = 0.99

2000

2000

Mean total weight (g)

Mean total weight (g)

The RAS described in this study was designed and operated

with the aim to investigate the growth of S. lalandi reared over
an extended period of time (488 days). The long-term sustainable operation of a modern aquaculture unit will increasingly
require the integration of cost-effective treatment process which
can include the re-use of water. The closed RAS with less than 1%
of water renewal rate per day, working with articial seawater,
allowed the investigation of system components and their interactions with specic biological, physical and chemical processes.
Taking this background into consideration, the type of RAS
under study needed special technical approaches in order to evaluate the performance of the system (water quality versus sh
growth performance) while aiming a low rate of water consumption. This aspect was considered as an operational key factor, often
limiting the commercial application in land-based sh farm facilities, because of (a) costs and (b) not providing the anticipated
benet for minimizing the environmental impact associated to the
discharge of wastes (including particulate and dissolved matter).
The design of the experimental RAS included a two-step solid
separation, a design element that differs from many other systems. Large particles were collected and eliminated via the drum
lter, while ne solids were removed via the protein skimmer.
The use of ozone in this study nds several comparative investigations (Blogoslawsky and Stewart, 1977; Brazil et al., 1998; Chen
et al., 1993; Colberg and Lingg, 1978; Honn, 1979; Kobayashi et al.,
1993; Krumins et al., 2001a; Rosenthal and Otte, 1979; Rosenthal
and Krner, 1985; Sander and Rosenthal, 1975; Schroeder et al.,

1500
1000
500
0

0
0

100

200

300
Days

400

500

10

20
30
Mean fork length (cm)

40

50

Fig. 7. Growth performance of Seriola lalandi (A) and length-weight relationship (B) in the RAS over 488 days of experimentation. Length-weight mathematical relationship
and correlation index is presented.

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J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028

25

Table 4
Mean weight, mean length and condition factor (K) (standard deviation), specic growth rate (SGR), tank biomass and stocking density, feeding rate, mortality and feed
conversion ratio (FCR) determined for 21 sample periods (N = 50 individuals in average) between day zero until day 488. Harvesting of sh is included in mortality calculations
from day 187 until day 488.
Day

Fish
number

Mean weight
(g)

Mean length (cm)

0
7
14
21
35
49
62
77
103
110
139
147
166
180
187
211
245
286
306
359
488

1461
1343
1306
1292
1283
1259
1257
1225
877
858
844
831
756
735
731
571
545
531
522
502
374

0.7 0.2
1.9 0.4
4.2 0.9
8.2 2.0
28.7 4.0
52.7 10.3
74.3 16.0
93.1 23.1
138.4 33.2
148.8 38.9
201.6 70.8
245.6 93.4
331.1 91.9
446.4 131.4
470.0 131.7
537.6 193.6
683.3 190.2
903.3 290.6
928.0 214.5
1280.6 256.9
2006.0 339.0

3.2
4.6
5.8
7.3
10.0
13.2
15.6
14.6
18.9
19.3
21.4
22.8
25.5
27.7
26.0
30.2
32.7
35.3
36.0
39.5
41.7

0.3
0.3
0.4
0.6
0.6
1.0
1.6
1.5
2.1
2.3
2.7
3.0
3.4
3.3
3.8
4.0
3.2
4.2
3.4
4.4
5.0

K
2.2
1.9
2.1
2.1
2.9
2.3
2.0
2.0
2.1
2.2
2.0
2.0
2.0
2.1
2.7
1.9
1.9
2.0
2.0
2.1
2.9

SGR
(%BW/d)

0.2
0.2
0.2
0.3
0.5
0.3
0.3
0.7
0.4
0.4
0.4
0.6
0.5
0.5
0.6
0.4
0.4
0.5
0.4
0.5
1.2

13.5
11.4
9.4
8.9
4.3
2.6
1.5
1.5
1.0
1.0
2.5
1.6
2.1
0.9
0.5
0.7
0.7
0.1
0.6
0.3

Biomass (kg)

Stocking density
(kg/m3 )

Feeding
rate (%BW)

1.08
2.56
5.54
10.62
36.88
66.29
93.37
114.04
121.34
127.71
170.17
204.08
250.36
328.13
347.61
306.95
372.42
479.67
484.43
642.84
750.24

0.1
0.4
0.8
1.5
5.1
9.1
12.9
15.7
16.7
17.6
23.5
28.1
34.5
45.3
47.9
42.3
51.4
66.2
66.8
88.7
103.5

8.1
8.4
9.0
6.8b
4.0
4.5
3.7
2.2
2.9
2.2
1.9
1.5c
1.5
1.7
1.3d
1.4
1.2e
0.9
0.8
0.6

Mortality
(%/d)

FCR

1.15f
0.39
0.15
0.05
0.13
0.01
0.17
1.09g
0.31
0.06
0.19
0.48
0.20
0.08
0.91h
0.13
0.06
0.08
0.07
0.20i

0.60
0.74
0.96
0.76
0.94
1.69
2.44
1.44
2.80
2.13
0.77
0.95
0.72
1.85
2.50
2.04
1.82
6.70
1.34
1.78

Changes in feed type/size:

a
Nutra/1.2 mm.
b
Marico Mistral/3 mm.
c
Europe 15/R-3.
d
Europe 15/R-5.
e
Europe 15/R-7.
f
High mortalities (N = 118) after stocking.
g
High mortalities (N = 283) due to a low oxygen event.
h
Transfer of individuals (N = 154) to another RAS facility.
i
Transfer of individuals (N = 81) to another RAS facility.

2011; Summerfelt, 2003; Tango and Gagnon, 2003) but not with an
identical system conguration. Ozone was applied to enhance the
formation of foam and to optimize particle removal (Chen et al.,
1993; Krumins et al., 2001b; Reuter and Johnson, 1995). Water
renewal rates of less than 1% of the system volume per day are one
of the outcomes that indicate the high performance characteristics
of the RAS conguration. Although often not practical in largescale systems, the overall attempt to minimize water exchange
has relevance in terms of judging the effectiveness of operational
conditions in the system.
The RAS and its components should offer save limits to the
species in cultivation, in terms of TAN, NO2 -N and NO3 -N concentrations, as have been tentatively identied in the scientic
literature (Wickins, 1980; Hochheimer and Wheaton, 1997; Malone
and de los Reyes, 1997) and by intergovernmental and regional
regulatory authorities (EIFAC, 1986). Excessive concentrations of
these nitrogen compounds may result in growth depletion (Guillen
et al., 1993; Tomasso, 1993; Wajsbrot et al., 1993), immune depression (Goncalves et al., 2012; Hurvitz et al., 1997; Li et al., 2013),
and death (Lemari et al., 2004; Meade, 1985; Person-Le Ruyet
et al., 1995; Russo and Thurston, 1991; Tomasso, 1993) depending
on concentration level and time of exposure. Compared to owthrough ponds, reuse systems typically have signicantly reduced
make-up ows that can result in the accumulation of unwanted
compounds. Also, the ability to independently control parameters
such as dissolved oxygen, may allow culture animals to tolerate
higher levels of other water quality parameters (compare with
other culture types) (Colt, 2006).
The performance of the aerobic biolter in the present work was
satisfactory in terms of the requirements for an intensive sh farm.
Relatively high TAN and NO2 -N concentrations were observed
at the beginning of the experiment. Similar observations were
reported by Collins et al. (1975), LaBomascus et al. (1987), Nijhof

and Bovendeur (1990), Olivar et al. (2000), Thoman et al. (2001),

and Seo et al. (2001). Average TAN and NO2 -N concentrations were
higher than those reported by Abbink et al. (2011), but within the
safe limits recommended for warm water intensive aquaculture
systems. Blancheton (2000) considered that TAN concentration
for sea bass adults should be below 2 mg/L, while reduced growth
performance was observed at values ranging between 6 and
10 mg/L (Lemari et al., 2004). It has to be noted that the effect
of ammonia concentration may have greatly varied because of
numerous other interfering factors that affect the dissociation of
the unionized ammonia fraction (NH3 ). The equilibrium between
unionized (NH3 ) and ionized (NH4 + ) ammonia is highly dependent
on water pH, salinity, and temperature. Although the average pH
in the present study was relatively low (7.5), the percentage of
NH3 at 22 C is about 2% (Wheaton, 1977). NO2 -N concentration
averaged 0.21 mg/L (max 2.4 mg/L). These values are slightly
higher than those reported by Abbink et al. (2011) for the same
temperature range, but concordant with the safe limits described
by Blancheton (2000) and Roncarti et al. (2006) for Dicentrarchus
labrax. Nitrite is the ionized form of the relatively strong acid,
nitrous acid (Colt, 2006). For a NO2 -N concentration of 1.0 mg/L
NO2 -N, the concentration of nitrous acid (HNO2 ) is 0.086 g/L at
a pH of 7.5. In contrast to nitrite, HNO2 is freely diffusible across
gill membranes. While the concentration of unionized nitrous
acid increases at lower pH, there is no evidence of increase nitrite
toxicity at lower pH values (Colt, 2006).
The NO3 -N concentrations for large marine sh should be less
than 500 mg/L (Pierce et al., 1993). Thoman et al. (2001) described
average values of 120.2 mg/L NO3 -N in 80 days of experiments.
In this study, the highest NO3 -N concentration measured was
154.5 mg/L, far below the limit reported by Pierce et al. (1993).
After the denitrication unit was installed, NO3 -N removal was
observed, keeping the concentration below 40 mg/L. Undoubtedly,

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26

J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028

the anaerobic process is suitable to remove nitrogen and partly

phosphorous effectively. However, in contrast to the nitrication, a
hydrogen and carbon donor must be added in a correct relation to
the inuent NO3 -N and oxygen ux. Otherwise, toxic compounds
like nitrite or hydrogen sulphide can be formed, affecting the sh
even in small concentrations. In order to minimize time and effort
regarding the correct dosage of the hydrogen and carbon donor,
ORP seemed to be a useful tool for a qualitative and quantitative
evaluation of the NO3 -N removal. The denitrication rate can be
estimated by using ORP as a surrogate indicator so that continuous
nitrogen removal in the biological denitrication processes can be
maintained (Cheng et al., 2012). The results presented in this study
differ partially with those reported by Lee et al. (2000) who found
ORP values for an optimal denitrication process in fresh water
between 375 mV and 400 mV. The ndings presented by Cheng
et al. (2012) showed nitrate reduction rates between 137 and
125 mV at pH 7.4, slightly higher to the upper limit (150 mV)
of the working range presented here. The moving-bed anaerobic
reactor proved to be very reliable and almost maintenance-free.
The mechanical stirring of the biomedia was sufcient, in order to
prevent clogging.
The growth performance data reported in this study can be
compared partly with limited results found in the literature. As
described by Brett (1979), temperature is one of the key abiotic
factors affecting growth. Abbink et al. (2011) found lower SGR
values than in this study for the same range of sh size cultured at
temperatures around the average value presented here (22.6 C).
The ndings of Pirozzi and Booth (2009) when measuring the
routine metabolic rate (RMR) of yellowtail kingsh at six temperatures, between 10 C and 32.5 C, showed the thermosensivity
of RMR in this species. The lowest temperature quotient (Q10 )
occurred between 20 and 25 C and the asymptotic value of the
regression between Q10 and temperature was at 22.8 C. On the
contrary, Abbink et al. (2011) found that growth was optimized
at a water temperature of 26.5 C. It has to be noted that water
temperature and the corresponding content of dissolved oxygen
(DO) has not only impact on growth performance, but also major
costs and operational implications for RAS cultivation. As reported
by Pirozzi and Booth (2009) it is critical that high DO levels are
maintained for yellowtail kingsh and preferable at saturated
levels. When considering that RAS can theoretically offer any level
of water quality parameters to determined species, it is important
to nd the economically optimal growth parameters, by accepting
e.g. lower growth rates at lower temperatures in order to optimize
water oxygenation costs. Fernandes and Tanner (2008) found SGR
in two commercial pens to be 0.25 and 0.32%BW/d in average, in a
grow-out period that lasted 324 and 288 days respectively. These
SGR data are slightly lower than those determined in this study for
the same sh size. In contrast, the growth performance presented
here is mainly lower than the high SGRs reported by Kolkovski
and Sakakura (2004) for sea cages culture conditions in South
Australia. Growth data obtained in this study are higher than those
summarized by Masumoto (2002) for Japanese yellowtail Seriola
quinqueradiata, when comparing the same range of temperature
and sh size.
Although there appear to be no data on FCR of yellowtail kingsh
cultivated in a closed RAS, from juvenile to 2 kg sh, the average FCR
of 1.75 reported here (1.49 when not considering the highest value
of 6.7 determined on day 306) is higher than the values reported by
Abbink et al. (2011) in a semi-closed RAS and by Moran et al. (2009)
in cages, but much lower than the FCRs reported by Fernandes and
Tanner (2008). Masumoto (2002) shows a FCR of 2.63 in a 112 days
trial with S. quinqueradiata reared from 365 g to 759 g, and a FCR
of 2.13 in an experiment lasted 182 days with Japanese yellowtail
cultivated between 1190 g and 3260 g, both results for sea cages.
Relatively high single FCR values cannot be explained, even after

a retrospect analysis of the system performance in terms of water

quality. Looking into the increase of stocking density towards the
end of the trial to values above 50 kg/m3 , and according to the
results presented by Roncarti et al. (2006) with European sea bass,
reduced growth performance should be observed. In contrast, a FCR
of 1.34 was achieved with a stocking density of 88.7 kg/m3 , differing
from the ndings of Roncarti et al. (2006) ndings.
The condition factor (K) is commonly used as an indicator of the
nutritional condition of the sh in both wild and cultured stocks;
however, in yellowtail kingsh farming, surprisingly few authors
use this factor when describing sh performance in their studies.
Froese (2006) identies K along with the determination of maturity
stages, as a useful management tool for RAS operators when cultivating species up to a relatively high market size. Although K is
species dependant (Roncarti et al., 2006), Kono et al. (2000) found K
values between 1.4 and 1.5 for the related species S. quinqueradiata
with body weights between 398 g and 677 g cultivated in cages. Furthermore, Montero et al. (2004) determined K values between 1.0
and 1.5 for amberjack (S. dumerili) reared in tanks at sizes between
250 g and 1023 g. Unfortunately, these data are difcult to compare
because sh were exposed to different environments which exhibit
much more variable (i.e. seasonal) feeding conditions than derived
under RAS culture conditions. Presumably, high content of adipose
tissue or fat in yellowtail kingsh reared in RAS will partly affect
the K, which have a negative repercussion and therefore may be
interpreted wrongly.
The commercial scale-up of the system presented here is not
trivial. Table 5 presents the production costs for three different bio-economic scenarios. The base case scenario describes the
production of yellowtail kingsh (500 mt/y) considering the scientic and practical experience gathering in this study. The average
and best case scenario considered improvements in the technology, which take place in the scale-up process. Therefore, these
assumptions are not validated by practical experience. The best
case scenario is based on following assumptions: (1) an increase
of the mean stocking density without affecting sh welfare and
Table 5
Production costs per kilogram of yellowtail kingsh (S. lalandi) for a medium-scale
RAS (500 mt/y) working with articial sea water and located in Germany, for three
different bio-economic scenarios.
Bio-economic assumptions

Base case

Average case

Best case

Stocking density (kg/m3 )

Survival rate (%)
Water renewal rate (%)
FCR

50
85
1.0
1.5

55
90
0.7
1.4

60
95
0.4
1.3

Operational costs
ex farm

D /kg
sh

Fingerlings
Feed
Sea salt
Tap water
Waste water
Electrical energy
Thermal energy
Liquid oxygen
Acetic acid
Lime

1.12
2.47
0.36
0.05
0.11
0.80
0.17
0.22
0.03
0.04

16.1
35.5
5.2
0.7
1.6
11.5
2.4
3.2
0.5
0.5

1.06
2.32
0.23
0.03
0.07
0.73
0.15
0.22
0.03
0.04

16.8
36.8
3.6
0.5
1.2
11.6
2.4
3.4
0.5
0.6

1.00
2.15
0.12
0.02
0.04
0.67
0.14
0.21
0.03
0.04

17.6
37.8
2.1
0.3
0.7
11.8
2.4
3.8
0.5
0.6

Variable costs
Personnel
Other expenses
Maintenance
Depreciation

4.76
0.37
0.56
0.36
0.91

68
5.3
8.0
5.2
13.1

4.31
0.33
0.51
0.33
0.83

68
5.3
8.0
5.2
13.1

3.88
0.30
0.46
0.30
0.75

68
5.3
8.0
5.2
13.2

Production costs
per kga

6.96

100

D /kg
sh

6.30

100

D /kg
sh

5.69

100

a
The production costs do not include slaughtering, processing, packing, distribution, marketing and nancing.

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J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028

using the same technology, (2) an increase of the survival rate due
to an optimization of the water quality, (3) a reduction in the water
renewal rate due to an optimization of integrated processes (IMTA),
and (4) an improvement in the FCR due to the use of feed specially designed for RAS operation, a better feed distribution, and an
optimization of sh grading.
5. Conclusions
The results presented in this study showed a satisfactory growth
performance for yellowtail kingsh S. lalandi in a closed RAS. Overall, feed conversion was acceptable from an aquacultural point of
view. The ndings on systems performance were within expected
values, considering the very low water renewal rate. This was especially evident considering the anaerobic denitrication process.
This is not well examined in seawater RAS culture. The system
design produced consistently good result over an extended period
of time. The water treatment scheme maintained optimal water
quality and secured animal welfare. The environmental sustainability of modern RAS does not rely on production results and/or
good water parameters only, but also on the optimization of the
use of land, energy, feed and water. Recent developments of IMTA
systems allow the use of RAS waste products as nutrients, coupling
different water loops with the main sh production water system.
Another possibility is the implementation of end of pipe treatments
such as articial wetlands. Taking into consideration that the future
development of marine aquaculture will face a paradigm shift, it is
important that a modern medium-scale (500 mt/y) urban RAS can
be able to deliver high quality sh and other aquaculture products
to niche markets in areas with high population density. A deeper
understanding of the interaction between nutrient inputs (feed),
nutrient retention (growth) and outputs (soluble and particulate
wastes) will help facing the sustainability of RAS and integrated
land based aquaculture.
Acknowledgements
We are grateful for the assistance and technical support given
by Valeriu Dewald. This work was possible thanks to the cooperation of Martin Sander, who provided the necessary equipment to
conduct the study.
References
Abbink, W., Garca, A.B., Roques, J.A.C., Partridge, G.J., Kloet, K., Schneider, O., 2011.
The effect of temperature and pH on the growth and physiological response of
juvenile yellowtail kingsh Seriola lalandi in recirculating aquaculture systems.
Aquaculture 330333, 130135.
Blancheton, J.P., 2000. Developments in recirculation systems for Mediterranean sh
species. Aquacultural Engineering 22, 1731.
Blogoslawsky, W.J., Stewart, M.E., 1977. Marine applications of ozone water treatment. In: Fochtmann, E.G., Rice, R.G., Browning, M.E. (Eds.), Forum on Ozone
Disinfection. International Ozone Institute, New York, pp. 266276.
Brazil, B.L., Summerfelt, S.T., Libey, G.S., 1998. Application of ozone to recirculating aquaculture systems. In: Libey, G.S., Timmons, M.B. (Eds.), Successes
and Failures in Commercial Recirculating Aquaculture. Conference Proceedings.
Northeast Regional Agricultural Engineering Service. July 1996, Ithaca, NY,
pp. 373389.
Brett, J.R., 1979. Environmental factors and growth. In: Hoar, W.S., Randall, D.J., Brett,
J.R. (Eds.), Fish Physiology. Academic Press, London, New York, pp. 599675.
Bullock, G.L., Summerfelt, S.T., Noble, A.C., Weber, A.L., Durant, M.D., Hankins,
J.A., 1997. Ozonation of a recirculating rainbow trout culture system. Part I:
effects on bacterial gill disease and heterotrophic bacteria. Aquaculture 158,
4355.
Chen, S., Timmons, M.B., Bisogni Jr., J.J., Aneshansley, D.J., 1993. Protein and its
removal by foam fractionation. The Progressive Fish-Culturist 55, 7682.
Cheng, H.-B., Kumar, M., Lin, J.-G., 2012. Interpretation of redox potential variation
during biological denitrication using linear non-equilibrium thermodynamic
model. International Biodeterioration & Biodegradation 67, 2839.
Colberg, P.J., Lingg, A.J., 1978. Effect of ozonation on microbial sh pathogens, ammonia, nitrate, nitrite, and BOD in simulated reuse hatchery water. Journal of the
Fisheries Research Board of Canada 35, 12901296.

27

Collins, M.T., Gratzek, J.B., Shotts Jr., E.B., Dawe, D.L., Campbell, L.M., Senn, D.R.,
1975. Nitrication in an aquatic recirculating system. Journal of Fish Diseases
32, 20252031.
Colt, J., 2006. Water quality requirements for reuse systems. Aquacultural Engineering 34, 143156.
EIFAC, 1986. Flow through and recirculation systems. Report of the working group
on terminology, format and unit measurements. In: European Inland Fisheries
Advisory Commission (EIFAC), FAO Technical Paper 49, 100 pp.
Fernandes, M., Tanner, J., 2008. Modelling of nitrogen loads from the farming of
yellowtail kingsh Seriola lalandi (Valenciennes, 1833). Aquaculture Research
39, 13281338.
Froese, R., 2006. Cube law, condition factor and weight-length relationships: history meta-analysis and recommendations. Journal of Applied Ichthyology 22
(4), 241253.
Goncalves, A.F., Pscoa, I., Neves, J.V., Coimbra, J., Vijayan, M.M., Rodrigues, P., Wilson,
J.M., 2012. The inhibitory effect of environmental ammonia on Danio rerio LPS
induced acute phase response. Developmental and Comparative Immunology
36, 279288.
Guillen, J.L., Endo, M., Turnbull, J.F., Hiroshi, K., Richards, R.H., Aoki, T., 1993.
Depressed growth rate and damage to cartilage of red sea bream associated with
exposure to ammonia. Bulletin of the Japanese Society of Scientic Fisheries 59,
12311234.
Hochheimer, J.N., Wheaton, F., 1997. Intensive culture of striped bass. In: Harrell,
R.M. (Ed.), Stripped Bass and Other Morone Culture. Elsevier Science, Amsterdam, The Netherlands, pp. 127165.
Honn, K.V., 1979. Ozonation as a critical component of closed marine system design.
Ozone Science and Engineering 1, 1129.
Hurvitz, A., Bercovier, H., van Rijn, J., 1997. Effect of ammonia on the survival and the
immune response of rainbow trout (Oncorhynchus mykiss Walbaum) vaccinated
against Streptococcus iniae. Fish & Shellsh Immunology 7, 4553.
Kobayashi, T., Yotsumoto, H., Ozawa, T., 1993. Closed circulatory system for mariculture using ozone. Ozone Science & Engineering 15, 311330.
Kolkovski, S., Sakakura, Y., 2004. Yellowtail kingsh, from larvae to mature
shproblems and opportunities. In: Cruz Surez, L.E., Ricque Marie, D., Nieto
Lpez, M.G., Villareal, D., Scholz, U., Gonzlez, M. (Eds.), Avances en Nutricin
Acucola VII Memorias del VII Simposium Internacional de Nutricin Acucola.
1619 Noviembre, Hermosillo, Sonora, Mxico.
Kono, M., Furukawa, K., Sagesaka, Y.M., Nakagawa, K., Fujimoto, K., 2000. Effect of
green tea extracts and green tea grounds as dietary supplements on cultured
yellowtail and ayu. Nippon Shokuhin Kagaku Kogaku Kaishi 47 (12), 932937.
Krumins, V., Ebeling, J., Wheaton, F., 2001a. Part-day ozonation for nitrogen and
organic carbon control in recirculating aquaculture systems. Aquacultural Engineering 24, 231241.
Krumins, V., Ebeling, J., Wheaton, F., 2001b. Ozones effect on power-law particle size
distribution in recirculating aquaculture systems. Aquacultural Engineering 25,
1324.
LaBomascus, D.C., Robinson, E.H., Linton, T.L., 1987. Use of water conditioners in
water-recirculation systems. Progressive Fish Culturist 49, 6465.
Lee, P.G., Lea, R.N., Dohmann, E., Prebilsky, W., Turk, P.E., Ying, H., Whitson, J.L.,
2000. Denitrication in aquaculture systems: an example of a fuzzy logic control
problem. Aquacultural Engineering 23, 3759.
Lemari, G., Dosdat, A., Coves, D., Dutto, G., Gasset, E., Person-Le Ruyet, J., 2004. Effect
of chronic ammonia exposure on growth of European seabass (Dicentrarchus
labrax) juveniles. Aquaculture 229, 479491.
Li, M., Chen, L., Qin, J.G., Li, E., Yu, N., Du, Z., 2013. Growth performance antioxidant
status and immune response in the darkbarbel catsh Pelteobagrus vachelli fed
different PUFA/vitamin E dietary levels and exposed to high or low ammonia.
Aquaculture 406407, 1827.
Malone, R.F., de los Reyes Jr., A.A., 1997. Categories of recirculating aquaculture systems. In: Timmons, M.B., Losordo, T. (Eds.), Advances in Aquacultural
Engineering. Aquacultural Engineering Society (AES) Proceedings III, ISTA, IV.
November 912, Orlando, FL. NRAES, New York, pp. 197208.
Masumoto, T., 2002. Yellowtail, Seriola quinqueradiata. In: Webster, C.E., Lim, C.E.
(Eds.), Nutrient Requirements and Feeding of Finsh for Aquaculture. CAB International, Wallingford, UK, pp. 131146.
Meade, J.W., 1985. Allowable ammonia for sh culture. The Progressive FishCulturist 47, 135145.
Montero, F.E., Crespo, S., Padrs, F., De la Gndara, F., Garca, A., Raga, J.A., 2004.
Effects of the gill parasite Zeuxapta seriolae (Monogenea: Heteraxinidae) on
the amberjack Seriola dumerili Risso (Teleostei: Carangidae). Aquaculture 232,
153163.
Moran, D., Pether, S.J., Lee, P.S., 2009. Growth, feed conversion and fecal discharge
of yellowtail kingsh (Seriola lalandi) fed three commercial diets. New Zealand
Journal of Marine and Freshwater Research 43, 917927.
Nijhof, M., Bovendeur, J., 1990. Fixed lm nitrication characteristics in sea-water
recirculation sh culture system. Aquaculture 87, 133143.
Olivar, M.P., Ambrosio, P.P., Cataln, I.A., 2000. A closed water recirculation system
for ecological studies in marine sh larvae: growth and survival of sea bass larvae
fed with live prey. Aquatic Living Resources 13 (1), 2935.
Person-Le Ruyet, J., Chartois, H., Quemener, L., 1995. Comparative acute ammonia toxicity in marine sh and plasma ammonia response. Aquaculture 136,
181194.
Pierce, R.H., Weeks, J.M., Prappas, J.M., 1993. Nitrate toxicity to ve species of marine
sh. Journal of the World Aquaculture Society 24, 105107.
Pirozzi, I., Booth, M.A., 2009. The routine metabolic rate of mulloway (Argyrosomus japonicus: Sciaenidae) and yellowtail kingsh (Seriola lalandi: Carangidae)

Author's personal copy

28

J. Orellana et al. / Aquacultural Engineering 58 (2014) 2028

acclimated to six different temperatures. Comparative Biochemistry and Physiology Part A 152, 586592.
Reuter, J., Johnson, R., 1995. The use of ozone to improve solid removal during
disinfection. Aquacultural Engineering 14, 123141.
Roncarti, A., Melotti, P., Dees, A., Mordenti, O., Angellotti, L., 2006. Welfare status of cultured seabass (Dicentrarchus labrax L.) and seabream (Sparus aurata
L.) assessed by blood parameters and tissue characteristics. Journal of Applied
Ichthyology 22, 225234.
Rosenthal, H., Otte, G., 1979. Ozonation in an intensive sh culture recycling system.
Ozone: Science & Engineering 1, 319327.
Rosenthal, H., Krner, G., 1985. Treatment efciency of an improved ozonation unit applied to sh culture situations. Ozone: Science & Engineering 7,
179190.
Russo, R.C., Thurston, R.V., 1991. Toxicity of ammonia, nitrite, and nitrate to shes.
In: Brune, D.E., Tomasso, J.R. (Eds.), Aquaculture and Water Quality. The World
Aquaculture Society, Baton Rouge, LA, pp. 5889.
Sander, E., Rosenthal, H., 1975. Application of ozone in water treatment for home
aquaria, public aquaria and for aquaculture purposes. In: Blogoslawski, W.J.,
Rice, R.G. (Eds.), Aquatic Applications of Ozone. International Ozone Institute,
Stamford, CT, pp. 103114.
Schroeder, J.P., Croot, P.L., Von Dewitz, B., Waller, U., Hanel, R., 2011. Potential
and limitations of ozone for the removal of ammonia, nitrite, and yellow substances in marine recirculating aquaculture systems. Aquacultural Engineering
45, 3541.
Seo, J.-K., Jung, I.-H., Kim, M.-R., Kim, B.J., Nam, S.-W., Kim, S.-K., 2001. Nitrication
performance of nitriers immobilized in PVA (polyvinyl alcohol) for a marine
recirculating aquarium system. Aquacultural Engineering 24, 181194.

Spotte, S., 1979. Seawater Aquariums: The Captive Environment. John Wiley and
Sons, New York, 413 pp.
Summerfelt, S.T., 2003. Ozonation and UV irradiation: an introduction and examples
of current applications. Aquacultural Engineering 28, 2136.
Summerfelt, S.T., Hochheimer, J.N., 1997. Review of ozone processes and applications as an oxidizing agent in aquaculture. The Progressive Fish Culturist 59,
94105.
Summerfelt, S.T., Hankins, J.A., Weber, A.L., Durant, M.D., 1997. Ozonation of a recirculating rainbow trout culture system. Part II: effects on microscreen ltration
and water quality. Aquaculture 158, 5667.
Tango, M.S., Gagnon, G.A., 2003. Impact of ozonation on water quality in marine
recirculation. Aquacultural Engineering 29, 125137.
Thoman, E.S., Ingall, E.D., Davis, D.A., Arnold, C.R., 2001. A nitrogen budget
for a closed, recirculating mariculture system. Aquacultural Engineering 24,
195211.
Tomasso, J.R., 1993. Toxicity of nitrogenous wastes to aquaculture animals. Journal
of Aquatic Animal Health 5, 6472.
Wajsbrot, N., Gasith, A., Diamant, A., Popper, D.M., 1993. Chronic toxicity of ammonia to juvenile gilthead seabream, Sparus aurata, and related histopathological
effects. Journal of Fish Biology 42, 321328.
Wheaton, F.W., 1977. Aquacultural Engineering. Robert E. Krieger Publishing Company, Malabar, FL, pp. 708.
Wickins, J.F., 1980. Water quality requirements for intensive aquaculture: a review.
In: Tiews, K. (Ed.), Proceedings of the World Symposium on Aquaculture in
Heated Efuents and for Recirculation Systems, vol. I. Stavanger, Norway, 2830
May 1980, pp. 1737.