Aquaculture 422423 (2014) 17

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

Growth, digestive activity, welfare, and partial cost-effectiveness of

genetically improved farmed tilapia (Oreochromis niloticus) cultured in a
recirculating aquaculture system and an indoor biooc system
Guozhi Luo a,b,c, Qi Gao a, Chaohui Wang a,b, Wenchang Liu a,b, Dachuan Sun a,b,c, Li Li a,b,c, Hongxin Tan a,b,c,
a
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
b
Shanghai Aquaculture Engineering Technology Research Center, Shanghai 201306, China
c
Shanghai University Knowledge Service Platform, Shanghai Ocean University Aquatic Animal Breeding Center (ZF1206), Shanghai 201306, China

a r t i c l e i n f o a b s t r a c t

Article history: An 87-d experiment was conducted to investigate the growth, digestive activity, welfare, and partial cost-
Received 2 August 2013 effectiveness of raising genetically improved farmed tilapia (GIFT) in a recirculating aquaculture system (RAS)
Received in revised form 18 November 2013 and an indoor biooc technology (BFT) system. The stocking density was 8.06 kg m3 in all tanks at the begin-
Accepted 20 November 2013
ning of the study, and reached 44.95 kg m3 in the BFT tanks and 36.87 kg m3 in the RAS tanks at the end of the
Available online 1 December 2013
study. The individual sh weight at harvest was 22% higher in the BFT sh than in the RAS sh. The total weight
Keywords:
gain and specic growth rate of the BFT sh were, respectively, 128% and 112% higher than those of the RAS sh.
Tilapia The feed conversion ratio for BFT was 18% lower than that for the RAS. There was no signicant difference in the
Biooc technology crude protein (CP) and crude lipid (CL) content from the sh back muscle between the sh in the RAS and the BFT
Recirculating aquaculture system tanks. The CP and CL contents of the BFT sh were 30.90 9.04% and 1.27 0.61%, respectively. The activities of
Growth performance lipase in the stomach and intestine showed substantial differences between the RAS and the BFT sh, whereas
Digestive enzyme activity the activity of protease did not. There was no signicant difference in the activities of alkaline phosphatase (ALP),
Welfare lysozyme (LYZ) of the hepatopancreas, head kidneys, and serum between treatments. The total superoxide dismut-
ase (T-SOD) activity of the serum of the BFT sh was considerably higher than that of the RAS sh. The accumula-
tions of ammonium nitrogen (highest: 60 0.45 mg L1) and nitrite nitrogen (highest: 119 2.01 mg L1)
were observed in the BFT tanks. The phosphate concentration in the BFT tanks (b 4.01 0.34 mg L1) was sub-
stantially lower than that in the RAS tanks. The sh survival rate was 100% for both the RAS and BFT. Partial analysis
of the cost-effectiveness of operations under experimental conditions revealed that the BFT model was more effec-
tive than the RAS in tilapia culture.
2013 Elsevier B.V. All rights reserved.

1. Introduction
The advantages of using a recirculating aquaculture system (RAS)
appeal to both the aquaculture industry and society (Kolarevic et al.,
2012). In RAS sh tanks, water is recycled by using waste treatment in-
frastructures, such as drum lters and biological lters, for controlling
and stabilizing the environmental conditions of the sh, reducing the
amount of water used, and improving sh welfare (Martins et al., 2010).
Currently, the biooc technology (BFT) used in aquaculture has re-
ceived considerable attention because using it involves high production
yields, feed protein recycling, water quality, and bacterial infection con-
trol (Avnimelech, 2006; Crab et al., 2007; Little et al., 2008). Detritivorous
Corresponding author at: Hucheng Ring Road 999, Shanghai Ocean University,
Shanghai 201306, China. Tel.:+86 21 61900413; fax: +86 21 61900402.
E-mail address: drizzles@163.com (H. Tan).
species such as shrimp and tilapia that are lter feeders are ideal for such
a system (Azim and Little, 2008). In practice, however, problems still exist
in using a BFT system to culture sh. Organic carbon must be added to
the water to maintain a C:N ratio of N10. Furthermore, implementing a
system for mixing and aerating the water, which raises energy costs, is re-
quired to maintain an active biooc in suspension and to meet the oxygen
demand of elevated water respiration (Avnimelech, 2012; Hargreaves,
2013). Particularly, the suspended solid content in BFT is typically greater
than 500 mg L1, and excessive solid concentrations can clog the gills of
sh or shrimp, which affects their growth and welfare.
The idea of conducting the current study is based on the reality that
both the BFT-aquaculture system and RAS model can be used to culture
sh in indoor closed water. If successful, BFT might be a promising alter-
native technology for a conventional RAS in cultivating both lter feeding
and detritivorous aquaculture species. An experiment was conducted
to evaluate genetically improved farmed tilapia (GIFT) welfare in BFT-
managed tanks, to compare dissolved inorganic nitrogen dynamics, to

0044-8486/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.aquaculture.2013.11.023
2 G. Luo et al. / Aquaculture 422423 (2014) 17
evaluate sh growth, to compare sh digestive activity, and to evaluate
the cost-effectiveness of using BFT and RAS in aquaculture.
2. Materials and methods
2.1. Experimental facilities
The experimental scale-RAS used in the current experiment was
equipped with three tanks (the volume of each tank was 1 m3, and
the working volume in the current experiment was 0.3 m3 of water),
one sand lter, and three biological lters (Fig. 1). The water in the
RAS was renewed 9.6 times per day because of a moderate ow-
through rate of approximately 101 min1. In the application of BFT,
only three tanks were used and the water treatment units were closed.
The working volume in each BFT tank was 0.3 m3.
2.2. Fish stocking and tank management
The water was continuously aerated using compressed air root
blowers to maintain the dissolved oxygen (DO) of N 6 mg L1 in both
the BFT and RAS tanks. One root blower (135 W) was used for the
three tanks in the RAS tanks. Considering the high suspended-solid con-
centration, the high oxygen consumption that was required by the
adding of the carbon sources, and the strong mixing intensity required
in the BFT tanks, two root blowers were used in the three BFT tanks.
The pH in both systems was maintained at a range of 7.07.5 using
NaHCO3. A water temperature-control device was used to maintain the
temperature of both systems between 24 and 26 C. Furthermore,
water was added daily to account for evaporation loss and biooc remov-
al, and a simulated natural photoperiod (12 L:12D) was applied. The ni-
trication function in biolters was already established in the RAS and
the biooc was already developed in the BFT tanks (mixed liquor
suspended solids (MLSS) N150 mg L1) before the tilapia were put in.
Mixed sex GIFT were obtained from a commercial sh farm and
reared at a recirculating aquaculture facility for 21 d to acclimatize to
the experimental environment. The experiment was conducted over a
period of 87 d. One hundred tilapia (average individual weight
24.17 2.49 g) were stocked at 8.06 kg m3 in each tank at the begin-
ning of the experiment. The sh were fed a commercial-pellet diet
(Shanghai Nong Hao Special Feed Co., Ltd., Shanghai, China) that
contained 11.0% water, 44% crude protein, 8.0% crude lipids, 4.5% Ca,
1.2% P, 1.8% lysine, 1.4% methionine, b 16.0% crude ash, and b5.0%
crude ber. The feeding rates were based on observations of the sh's
feeding behavior during the adaptation period and xed at 2.0% of the
total stocking biomass daily, and adjusted fortnightly after weighing
the sh samples every 10 d (Azim and Little, 2008). The daily feed
rations were split into three equal amounts given at 0830, 1230, and
1830 to each tank. Sodium acetate was added at the rate of 75% of the
feed and was applied to the BFT tanks to maintain an optimum C:N
Fig. 1. Schematic diagram of the experimental-scale recirculating aquaculture system.
1 tank, 2 sand lter, 3 temperature control device, 4 biolters.
ratio for bacteria (Gao et al., 2012). The biooc uptake of the sh was in-
sufcient to prevent its accumulation; therefore, the biooc required
regular removal from the system to maintain the total suspended solids
(TSS) at less than 500 mg L1. Whenever the MLSS level exceeded
500 mg L1 in any BFT tank, some of the water containing ocs was
removed. After settling for 1 h, the supernatant was taken back to the
tank and the biooc was collected and stored below 20 C.
2.3. Methods of determining experimental parameters
2.3.1. Water quality parameters
The water temperature, DO, and pH were measured daily between
0830 and 0930 by using a YSI-556 meter (YSI Incorporated, Yellow
Springs, OH, USA). The MLSS, phosphate (PO34 -P), total ammonium ni-
trogen (TAN), nitrite (NO
2 -N) and nitrate (NO3 -N) were analyzed
using a standard method (APHA, 1998). The MLSS and TAN were mea-
sured every day, and the NO 3
2 -N, NO3 -N and PO4 -P were measured
every 3 d. The dissolved organic carbon was evaluated daily using a
total organic carbon analyzer (TOC-V CPH; Shimadzu Seisakusho,
Japan).
2.3.2. Fish digestive activity and welfare indicators
The sh were starved for 24 h before sampling for digestive activity
and welfare indicators at the end of the experiment. Ten sh were
randomly netted from each tank and immediately anesthetized using
2-phenoxyethanol (1:300 v.v1) in a 60 L bucket. The sh reached
stage V of anesthesia within 23 min and were subsequently stunned
by a blow to the back of the head. Blood was collected from the caudal
vein using a 1 mL syringe with a 22 G 1-1/2 in. needle within 1 min
of sampling. The time from capture to blood collection was b5 min.
Each blood sample was placed in a 1.5-mL nonheparinized Eppendorf
tube. After clotting, the samples were centrifuged at 4000 rpm for
10 min at 4 C. The serum samples were split into several aliquots and
stored at 20 C, and analyses were performed on the following day.
The digestive stomachs and intestines, hepatopancreases, and head
kidneys of the sampled sh were taken out and homogenized three
times using normal saline solution (1:3 w/v) at 0 C for 30 s each
time, using an electric blender operating at 8000 rpm. The homogenate
was centrifuged at 10,000 g, 4 C for 30 min to eliminate tissue debris
and lipids. The supernatant (enzyme extract) was dispensed into 1.5-
mL Eppendorf tubes and kept at 20 C until enzymatic assay. The
back muscle was excised rapidly from the sh, frozen in liquid nitrogen,
and homogenized in nine volumes of a 20-mM phosphate buffer with a
pH of 7.4, 1 mM EDTA, and 0.1% triton X-100 at a temperature between
0 and 4 C. The homogenates were centrifuged at 600 g below 0 C to
remove debris, and the resultant supernatant uids were used directly
for the proximate composition analysis of sh muscle.
The protease activity was assayed according to the method of Lowry
et al. (1951), using casein as the substrate and using a Folin phenol
reagent to obtain a reaction. The protease enzyme activity was mea-
sured as the change in absorbance using an ELISA analyzer (Bio-Tek
ELX-750, Bio-Tek Instruments, Inc., Winooski, VT, USA) and expressed
as 1 g of tyrosine or maltose released per min (U g1 protein). One
unit of enzyme activity was expressed as 1 M tyrosine released per
min. Lipase activity was determined based on the measurement of
fatty acid release caused by the enzymatic hydrolysis of triglycerides
in a stabilized emulsion of olive oil (Borlongan, 1990; Jin, 1995). Enzyme
activities, including those of protease and amylase were both expressed
as specic activity U mg1 of protein, and lipase activity was expressed
as U g1 of intestinal content (Yanbo and Zirong, 2006).
The activities of serum LYZ were measured using the turbidimetric
assay developed by Parry et al. (1965) with the microplate adaptation
of Hutchinson and Manning (1996) using a kit. Serum ALP activity
was assayed using spectrophotometry as the release of p-nitrophenol
from the model substrate p-nitrophenyl phosphate according to Hadas
and Pinkas (1997) and Wang and He (2009). The T-SOD (EC1.15.1.1)
 G. Luo et al. / Aquaculture 422423 (2014) 17
activity in solution was determined using the method of Marklund and
Marklund (1974) based on the autoxidation of pyrogallol and modied
by Jing and Zhao (1995). The serum glucose concentrations were
measured using a biochemical analyzer (Mindray Chemistry Analyzer
BS-200; Shenzhen, Guangdong Province, China), and the kits used
were supplied by Mindray Medical International Limited (Shenzhen,
China).
2.3.3. Biochemical composition of the oc and sh muscle
Every 3 d, the oc produced in BFT tanks was collected. After being
settled and concentrated, biooc samples were dried in an oven at
105 C to constant weight and then subjected to a proximate composi-
tion analysis. The crude protein and crude lipid content of the biooc
sample and sh muscle sample were determined by performing the
standard methods GB/T6432-94 and GB/T6433-94. The crude protein
content was determined by measuring nitrogen (N, 6.25) and using
the Kjeldahl method, and the crude lipid content was determined
using ether extraction with a Soxhlet extractor.
2.3.4. Fish growth parameters
At the end of the experiment, the sh were harvested and 10 sh
were collected from each tank to measure the nal body weight. The
observed body weight and food intake data were used to calculate the
following growth indices:
1) Weight gain (%) = (W2 W1) / W1
2) Specic growth rate (SGR) = [(lnW2 lnW1) / (t2 t1)] 100%
3) Feed conversion ratio (FCR) = feed supply (kg) / sh biomass
increase (kg)
4) Survival rate (%) = 100 (Nf Ni) / Ni
5) Total weight gain (kg/m3) = (W2 W1) / 0.3
Where W1 and W2 are the weights at times t1 and t2, respectively,
with t1 and t2 being the rst and nal day of the experiment, respectively,
n is the number of sh, F is the weight of total feed (g), Ni is the initial
number of sh, and Nf is the nal number of sh.
2.4. Statistical analysis
Statistical analysis was performed using SPSS 16.0 for Windows. The
differences in water quality were considered signicant when P b 0.05.
One way analysis of variance was used on the experimental parameters
including growth parameters, digestive activity, welfare indicators, and
production costs of BFT and the RAS.
3. Results and discussion
3.1. Water quality
During the experimental period, the water temperature was main-
tained at 2426 C, DO was maintained at over 6 mg L1, and pH was
maintained at 7.07.5 for all tanks.
In the BFT system, increasing the C:N ratio and maintaining a high
DO (N 6 mg L1) in the culture water enabled the assimilatory activity
of heterotrophic bacteria to convert ammonium into bacterial biomass.
Employing this process is more efcient than nitrication (in which am-
monium is nitried to NO 3 -N), maintains the stability of water quality,
and maintains oxygen and alkaline levels. Moreover, the existence of
the biooc created a heterogeneous microenvironment that led to
more possibilities for nitrogen recycling in the BFT system. Observing
nitrication occurring in BFT tanks is not unusual. NO
2 -N and NO3 -N
are produced through nitrication; in the BFT system, the produced
NO 3 -N might undergo incomplete denitrication to produce NO2 -N,
and dissimilatory NO 3 -N reduction to ammonia (NH 3 ) might also
occur under the experimental conditions (Wu et al., 2012). Therefore,
the nitrogen syndrome in BFT is complex and dynamic.
3
The obvious accumulation of NO 3 -N (70100 mg L
1
) and the
nearly undetectable NO 2 -N level (b 0.97 mg L
1
) in the RAS tanks indi-
cated that the biological lters function effectively (Fig. 2) in the exper-
imental RAS. By the nal period of the study, the sh density in the
culture tanks reached 36.87 kg m3, an increase of 360% from the initial
stocking density, which caused the TAN accumulation in the RAS tanks.
In the BFT tanks, the TAN was primarily from the decomposition of
organic matter such as sh waste and dead microbes in the reactors.
The TAN can be rapidly taken up and stored by the biooc microbes
by adding organic carbon to stimulate the growth of heterotrophic
bacteria. However, the packaging of nitrogen in microbial cells is tempo-
rary because cells turn over rapidly and release nitrogen as NH3 when
they decompose. Therefore, the TAN was repeatedly cycled between
the dissolved NH3 and the oc solids, exhibiting an obvious uctuation
(Fig. 2A). The peaks of the TAN and NO 2 -N occurred on day 20 during
the BFT system start-up. The highest concentrations of TAN and NO 2 -
N were 60 0.45 mg L1 (Fig. 2A) and 119 2.01 mg L1 (Fig. 2B),
respectively. With the acclimation of the system, the TAN and NO 2 -N
started to decrease. At the end of the study, there were no detectable
NO 2 -N, and the TAN was less than 10 mg L
1
. No apparent accumula-
tion of NO3 -N was observed in the culture tanks during the entire
experiment (Fig. 2C).
The safe level of NH3 for larval tilapia (Oreochromis niloticus) is
0.42 mg L 1 (Benli and Kksal, 2005). When the NH3 fraction from
the TAN at an overall average pH of 7.11 and average temperature of
22 0.15 C, approximately 0.5% are accounted for because the safe
TAN level for Nile tilapia is 84 mg L1, which is higher than the highest
TAN in the current study. Therefore, it can be concluded that our BFT
system is NH3 and toxicity-free.
NO 2 -N is an intermediate during both nitrication and denitrication
(Chuang et al., 2007; Ruiz et al., 2003). The accumulation of NO 2 -N is
common in intensive aquaculture (Wang et al., 2004), probably because
of the free NH3 inhibition during nitrication and denitrication (Shi
et al., 2011). Based on a recommendation from Wuertz et al. (2013),
salt was added to the BFT tanks (1%, w/w1) to compensate for the stress
from NO
2 -N to the tilapia in the current experiment. The NO2 -N concen-
tration in the BFT tanks was much higher than that from other reports
(Asaduzzaman et al., 2009, 2010; Xu and Pan, 2012), which is probably
because of the higher stocking density in the current experiment.
There is no accumulation of NO 3 -N in the BFT tanks perhaps because
the process of nitrication might be inhibited by the high C:N ratio,
the NO 3 -N was used by the heterotrophic bacteria (Luo et al., 2013),
or denitrication occurred in the system.
Previous studies on phosphorus dynamics in freshwater aquaculture
systems have revealed that much of the phosphorus added with the
feed is unutilized by the cultured sh and that a relatively large fraction
(80%90%) is released into the sh culture systems (Barak et al., 2003).
A targeted method for phosphorus removal in the current RAS was not
developed; therefore, the accumulation of PO3 4 -P in the RAS tanks was
45 1.23 mg L1 at the end of the experiment (Fig. 2 D), whereas the
1
PO3
4 -P level in the BFT tanks was as low as 4.01 0.34 mg L , which
was probably because the growth and reproduction of the biooc
microorganisms assimilated the phosphorus that was not used by the
sh. Moreover, some biooc was eaten by the sh, the phosphorus in
the biooc could be assimilated much easily than that in the feed, and
another portion of the oc was collected for biooc sampling in the
current study. Nearly all of the PO3 4 -P in the BFT tanks were depleted,
suggesting that using the BFT system improved both the nitrogen and
phosphorus usage efciency of feeding.
3.2. Growth performance and feed utilization of sh
The sh yield parameters are given in Table 1. Even when the water
quality was not stable and there were high uctuations in the concen-
trations of TAN and NO 2 -N, no sh died during the entire experiment,
and the sh in the BFT tanks grew more satisfactorily than those in
4 G. Luo et al. / Aquaculture 422423 (2014) 17

A TAN-RAS B Nitrite-RAS
TAN-BFT Nitrite-BFT
70 140

Concentrations (mg L-1)

Concentrations (mgL-1)
60 120
50 100
40 80
30 60
20 40
10 20

0 10 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90

C D

Concentrations of Phosphate (mg L-1)

Nitrite-RAS Phosphate-RAS
350 60
Nitrite-BFT Phosphate-BFT
Concentrations (mg L-1)

300 50

250 40
200
30
150
20
100
10
50

0 10 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90
Times (day) Times (day)

Fig. 2. Nitrogen and total phosphate dynamics in the RAS and BFT tanks. RAS: recirculating aquaculture system; BFT: biooc technology.

the RAS tanks did. The individual sh weight at harvest was 22% higher Yuan et al., 2010). The nal biomass level of 44.95 kg m3 in the current
in the BFT sh than in the RAS sh. The total weight gain and SGR of the BFT tanks were higher than the 1028 kg m3 achieved in indoor and
BFT sh were, respectively, 129% and 12% higher than those of the RAS outdoor BFT systems reviewed by Little et al. (2008). The RAS in the cur-
sh. The FCR for the BFT sh was 1.20 0.03, 18% better than that of rent study was set up on laboratory scale, and the water was aerated. The
the RAS sh, which had an FCR of 1.47 0.02. There was no substantial water exchange rate is 9.6 times per day in the RAS. During the 87-d ex-
difference in the crude protein and crude lipid content of sh back mus- perimental period, the total weight gain rate in the RAS sh was
cles between the sh BFT and the RAS sh (Table 2). 28.87 kg m3, equivalent to over 110 kg m3 per year, which is close
Previous studies have conrmed that biooc substantially contributes to the ndings of a previous report on tilapia cultured in a commercial-
to the growth and production of tilapia, which are known to use in situ- scale RAS (Timmons et al., 2010).
produced food particles such as suspended bacteria (Avnimelech, 2007;
Azim and Little, 2008; Beveridge and Baird, 2000; Little et al., 2008;
3.3. Part nutritional quality of biooc

It was suggested that diets containing 2530% crude protein and

Table 1
512% crude lipid was suitable for tilapia (Chou and Shiau, 1996;
Growth performance and feed utilization of tilapia in recirculating aquaculture system and Jauncey, 2000). The average crude protein content of biooc was
indoor biooc technology system. 30.90 9.04% (on a dry-matter basis) in the current experiment,
which was appropriate for tilapia (Table 3). However, the crude lipid
Parameters RAS BFT
content of biooc was only 1.27 0.61%, much lower than that of com-
Initial mean weight (g sh1) 24.17 2.49a 24.17 2.49a
mercial feed. The low crude lipid content was also reported in other
Initial number (sh tank1) 100 100
Initial stocking density (kg m3) 8.06 8.06 studies; for example, there was 3% crude lipid content of biooc in the
Final mean weight (g sh1) 138.29 34.61a 168.58 40.64b study of Azim and Little (2008).
Final number (sh tank1) 80 80
Survival rate (%) 100%a 100%a
Final density (kg m3) 36.87a 44.95b Table 2
Specic growth rate (% day1) 1.90 0.30a 2.13 0.29b Proximate compositions (% dry basis) of back muscle of tilapia cultured in recirculating
Weight gain (%) 472.14 143.18a 597.48 168.16b aquaculture system (RAS) and biooc technology (BFT) tanks.
Total weight gain (kg m3) 28.87a 36.95b
Compositions of muscle RAS BFT
Food conversion rate (kg kg1) 1.47 0.02a 1.20 0.03b
Crude protein 79.72 0.74a 80.25 1.86a
Note: Each value represents mean S.D. (n = 30). Values in the same row with different
Crude lipid 4.69 0.32a 4.23 1.04a
superscript letters are signicantly different (P b 0.05). 10 sh were taken out from each
tank on day 30 day and day 60. Therefore, at the end of the experiment, the sh number in Note: Each value represents mean S.D. (n = 30). Values in the same row with different
each tank was 80. RAS: recirculating aquaculture system. BFT: biooc technology. superscript letters are signicantly different (P b 0.05).
 G. Luo et al. / Aquaculture 422423 (2014) 17 5

Table 3 Table 5
Proximate composition of biooc, commercial feed and sh feces. Partial economic performance of tilapia cultured in recirculating aquaculture system
(RAS) and biooc technology (BFT) tanks for 87-day period.
Crude protein (%) Crude lipid (%)
ITEM RAS BFT
Feed 43.57 0.51a 6.51 0.31a
Feces 12.07 0.62c 0.99 0.18b Feed consumption (kg feed kg sh1) 1.47 1.20
Floc 30.90 9.04b 1.27 0.61b Energy for aeration (kw kg1) 11.22 22.44
Pumping for RAS (kw kg1) 12.47 0
Each value represents mean S.D. Values in the same row with different superscript
Water consumption (m3 kg1) 1.00 1.67
letters are signicantly different (P b 0.05).
NaHCO3 consumption kg kg1 weight gain 0.60 0.30
Carbon consumption kg glucose kg1 weight gain 0 0.90
a
Depreciation for water treatment unit (kg weight gain)1 9.61 0

A B Note: Each value represents mean S.E. Values in the same row with different
Intestinal Protease

a a superscript letters are signicantly different (P b 0.05).

2500 a b
Activity (U g-1)

Intestinal Lipase
a
Activity (U g-1)
20 5-year depreciation period.
2000
15
1500
1000 10
activities. Because these microbial enzymes can help break down pro-
500 5
teins, carbohydrates, and other nutritional ingredients in the feed,
0 0
RAS BFT biooc could presumably facilitate feed digestibility and absorption.
RAS BFT
Therefore, biooc can supplement microbial nutrition and contribute
C D to improving the digestion and utilization of the feed.
Stomach Protease

a
a b
Activity (U g-1)

600
activeity (U g-1)

70
Stomach lipase

500 60 a 3.5. Fish welfare

400 50
300 40
200 30
367.86 20
100
10
0
0 kidneys of the sh cultured in BFT and the RAS at the end of the exper-
RAS BFT RAS
Fig. 3. Specic activity of protease and lipase in the stomachs and intestines of tilapia
raised in RAS and BFT tanks at the end of the 87-d experiment. Means S.E. are indicated;
n = 30; and means within the same tissue with different letters are signicantly different
(P b 0.05). RAS: recirculating aquaculture system; BFT: biooc technology.
Perhaps the nutritional value of biooc could not be evaluated
simply by comparing its crude protein and crude lipid contents with
that of commercial feed. Biooc can be a complete source of cellular
nutrition and various bioactive compounds, and may contain some un-
discovered factors (Ju et al., 2008). With the same crude protein content
as commercial feed, biooc is more benecial for sh growth. Further
study is required to analyze the nutritional value of biooc by evaluating
the growth and immunity of the feeders.
3.4. Digestive activity of sh
The intestine lipase activity was signicantly lower in sh growing
in the BFT tanks than in those in the RAS tanks (P b 0.05), whereas
the opposite was true for stomach lipase activity. There was no signi-
cant difference in the protease activity in both the intestine and stomach
of the sh cultured using the two systems (P b 0.05) (Fig. 3).
One possible reason for this is that the lipid content in biooc is
much lower than the lipid requirements of tilapia, whereas the biooc
protein content fullls the sh's requirements. According to Xu and
Pan (2012), biooc exhibited relatively high protease and amylase
In the current study, there was no signicant difference in the ALP,
LYZ, or T-SOD activity (P N 0.05) in the hepatopancreases or head
BFT iment. Moreover, no signicant differences in the LYZ activity in the he-
patopancreas, serum ALP activity, and serum glucose concentrations
were found in the sh from BFT and the RAS (P N 0.05). However, the
serum T-SOD concentration in the sh cultured in BFT was substantially
higher than that in the sh cultured in the RAS (Table 4). SODs are a
group of metalloenzymes that play a key role in the defense against
the toxic effects of reactive oxygen species and can be useful
for assessing oxidative stress (Lenartova et al., 1997; Pedrajas et al.,
1998). In the current study, the tilapia cultured in BFT tanks exhibited
superior T-SOD activity, indicating that feeding on biooc improved
the immunity of the tilapia; this nding is consistent with the results
of the growth performance and digestive activity analysis.
3.6. Consumption of water, energy, and feed
Considering that the current study was designed on a laboratory
scale, only the cost of water, electricity, and feed, among others, during
the experimental period was used to compare the cost of running the
two types of systems(Table 5).
Because both of the culture models were closed systems, the water
consumption for production per kilogram of sh was low for both
systems. The consumption in the RAS tanks was lower because there
was some water loss in the BFT tanks when the biooc was taken out.
In the RAS, a water treatment unit and a pump for recycling water
were added; therefore, the related energy cost and depreciation cost
of the equipment was higher compared with that of BFT.
Carbon sources were added to the BFT tanks regularly to maintain a
certain C:N ratio and the cost for carbon sources was one of the major

Table 4
Performance of sh welfare of tilapia cultured in BFT and RAS at the end of 87-day experiment.

Alkaline phosphatase (U g1) Lysozyme activity (U g1) Total superoxide dismutase (U g1) Glucose concentrations (mmol1)

RAS BFT RAS BFT RAS BFT RAS BFT

Hepatopancreas 44.35 10.06a 53.125 11.05a 25.11 10.48a 19.98 7.99a 66.72 16.72a 46.20 10.72a
Head kidney 30.21 7.03a 37.95 4.84a 25.34 6.87a 25.37 6.34a 4.89 1.47a 5.19 0.95b
Serum 24.96 5.19a 21.09 1.64a 7.42 1.65a 8.05 2.05a 39.59 14.03a 43.82 8.37b 6.97 2.83a 6.74 0.69a

RAS: recirculating aquaculture system. BFT: biooc technology. Each value represents mean S.D (n = 30). Values in the same row with different superscript letters are signicantly
different (P b 0.05).
6 G. Luo et al. / Aquaculture 422423 (2014) 17
expenses in running a BFT system. In the current study, the amount of
carbon added to the BFT tanks was calculated based on the feeding
rate of the sh, and 0.9 kg of sucrose was consumed per kilogram of
sh produced.
In addition to carbon source, aeration is another major expense in
running a BFT system. Aeration is used to supply oxygen and mix the
components in the water (Hargreaves, 2013). In the current study,
one compressed air root blower (135 W) was used to maintain the
DO in the RAS tanks at over 6 mg L1. However, using two compressed
air root blowers was required in the BFT tanks to maintain the same DO
level (N6 mg L1). Approximately 11.22 kW of energy was consumed
to produce 1 kg of sh using RAS tanks, whereas 22.44 kW was required
to produce the same amount using BFT tanks.
In the process of nitrication, 7.15 g of alkaline was consumed per
gram of TAN converted to NO 3 -N in the RAS, whereas in the BFT, TAN
was assimilated by microorganisms and converted into biooc protein
and only 3.57 g of alkaline was consumed per gram of TAN converted
(Ebeling et al., 2006). During the experimental period, the amount of
NaHCO3 used to maintain a stable pH was less in the BFT tanks than in
the RAS tanks.
Some of the biooc produced in the BFT tanks were collected and
stored. Dried biooc has been used as an ingredient to replace shmeal
and soybean protein in shrimp feed (Azim and Little, 2008).
Because both systems were operated under laboratory conditions,
the amount of electricity used per kilogram of sh biomass gain was
great; therefore, using the same amount is perhaps impractical for even-
tually scaling up such a system. Thus, these values must be altered when
these systems are used in actual production.
4. Conclusion
The current study investigated the growth, digestive activity,
welfare, and partial production costs of raising GIFT in RAS tanks and in-
door BFT tanks. At the beginning of the 87-d culture period, the sh den-
sity was 8.07 kg m3 in both tanks; but at the end of the experiment,
the sh density was 44.95 kg m3 and 36.87 kg m3 in the BFT and
RAS tanks, respectively. The tilapia ourished in both environments;
however, the BFT sh were superior to the RAS sh, and no mortality
was found among the BFT sh, although the accumulation of NO 2 -N
and TAN was observed in the tanks. The crude protein content in the
biooc met the nutritional requirements of tilapia, but the crude lipid
concentration was quite low. A signicant difference was found for li-
pase in the intestines and stomachs of the sh cultured using both
models; however, no substantial difference was found for protease.
The serum T-SOD levels of the sh grown in the BFT tanks were signif-
icantly higher compared with the levels of those grown in the RAS
tanks; moreover, the BFT sh exhibited superior immunity to the RAS
sh. The partial production-cost analysis revealed that it is more cost-
effective to culture tilapia using BFT than using RAS.
Acknowledgment
This study was funded by Project 31202033 supported by the
National Natural Science Foundation of China, the Twelve Five National
Science and Technology Support Program (2012BAD25B03), and the
Project of Shanghai Engineering and Technology Center for Promoting
Ability (13DZ2280500).
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