Chemosphere 182 (2017) 8e14

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Biooc technology application in indoor culture of Labeo rohita

(Hamilton, 1822) ngerlings: The effects on inorganic nitrogen control,
growth and immunity
Dibyendu Kamilya a, *, Mitila Debbarma a, Prasenjit Pal b, Biswanath Kheti a,
Sudipto Sarkar c, Sukham Tushiba Singh a
a
Department of Aquatic Health and Environment, College of Fisheries, Central Agricultural University, Lembucherra, 799210, Tripura (w), India
b
Department of Extension and Social Science, College of Fisheries, Central Agricultural University, Lembucherra, 799210, Tripura (w), India
c
Department of Agricultural Engineering, Triguna School of Technology, Assam University, Silchar, 788011, Assam, India

h i g h l i g h t s

A biooc based aquaculture system with molasses as carbon source and Labeo rohita as culturable animal was developed.

Ammonia immobilization by heterotrophic bacteria was the dominant mechanism inuencing the inorganic nitrogen removal.

There were enhanced growth and survival of L. rohita in biooc system.

An indication of immunity enhancement of L. rohita was also observed in the biooc system.

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

Article history: The present study was undertaken to investigate the effect of biooc-based aquaculture system on
Received 4 January 2017 inorganic nitrogen control, and growth and immunological responses of rohu (Labeo rohita). Fingerlings
Received in revised form of rohu were cultured in indoor biooc tanks in light-limited condition for 16 weeks with daily addition
2 May 2017
of molasses as carbon source to maintain the C/N ratio at 15. Different water quality parameters were
Accepted 3 May 2017
Available online 3 May 2017
measured at weekly interval. Growth, survival and welfare indices including stress and immunological
status were measured after the end of the experimental period. Factor analysis using water quality
Handling Editor: A Adalberto Noyola variables indicated ammonia immobilization by heterotrophic bacteria, rather than nitrication, as the
dominant mechanism inuencing ammonia removal in the biooc system. Ammonia immobilization was
Keywords: concomitant with microbial biomass production, as also revealed through factor analysis. The biooc
Biooc culture system enhanced the growth of rohu signicantly than the control sh. As a measure of stress,
Aquaculture the serum glucose content did not differ signicantly between the sh cultured in biooc and control
Nitrogen removal system. Among the immunological parameters, respiratory burst and alkaline phosphatase activity
Factor analysis
showed signicant enhancement in sh grown in biooc system than the control. However, myeloper-
Growth
oxidase activity and total serum protein content did not show any statistically signicant difference
Fish welfare
between biooc and control system. The enhanced growth and welfare of the cultured sh may be
attributed to in situ maintenance of water quality within the biooc system and presence of microbial
oc.
2017 Elsevier Ltd. All rights reserved.

1. Introduction without hampering environmental, economic and social sustain-

One of the novel technologies that have the potential to
generate high production from intensied aquaculture system
* Corresponding author.
E-mail address: dibyendu.kamilya@gmail.com (D. Kamilya).
ability is biooc technology (De Schryver et al., 2008; Crab et al.,
2012). Originally described as activated suspension technique and
developed during the eighties, this technology is based on the
maintenance of high levels of microbial oc in suspension by
continuous aeration (Avnimelech et al., 1986; Sering, 2006).
Constant aeration allows aerobic decomposition of organic matters

http://dx.doi.org/10.1016/j.chemosphere.2017.05.021
0045-6535/ 2017 Elsevier Ltd. All rights reserved.
 D. Kamilya et al. / Chemosphere 182 (2017) 8e14
viz. feed, fertilizes, faeces etc. in an aquaculture pond stocked with
sh with the consequent development of a dense population of
heterotrophic microorganism (Avnimelech, 1999). Such dense and
suspended heterotrophic population controls water quality, be-
comes a protein rich food source for sh and can act as an alter-
native measure for pathogen control (Hargreaves, 2006; Crab et al.,
2007; Avnimelech, 2012).
Theoretically, the system operates through the addition of a
carbon source as a fertilizer to increases the C/N ratio which, in
turn, enhances conversion of inorganic nitrogen to microbial
biomass (Avnimelech, 1999). Micro-organisms utilize carbohydrate
as energy source to produce new cells and nitrogen is utilized for
the synthesis of protein which is also a major component in the
formation of new cells (Avnimelech, 1999). Thus, as a basic micro-
bial process, utilization of carbohydrate is accompanied by the
immobilization of inorganic nitrogen (Avnimelech, 1999). A high C/
N ratio (10e20) is recommended for development of biooc and
efcient ammonia removal (Hargreaves, 2006; Avnimelech, 2012).
This can be achieved by adding different locally available carbon
sources and/or using low nitrogen feed (Avnimelech, 1999;
Hargreaves, 2006).
The benets of biooc technology over conventional practices in
terms of water usage efciency and dynamic changes in water
quality have been successfully demonstrated in shrimp farming
(Avnimelech, 1999; Burford et al., 2003; Zhao et al., 2012; Xu et al.,
2016) and to some extent in nsh culture (Sering, 2006; Perez-
Fuentes et al., 2016). Owing to the limited work on biooc tech-
nology application in nsh aquaculture, more researches need to
be performed to include potential species under such system. Apart
from being lter feeder (Chondar, 1999) and periphyton feeder
(Azim et al., 2002), rohu (Labeo rohita) is also capable of ingesting
bacteria in suspension (Rahmatullah and Beveridge, 1993). These
attributes can make this species suitable for cultivation in biooc-
based system. However, biooc technology application for culture
of this species is yet to be fully demonstrated. Moreover, in general,
limited studies have investigated the potential of biooc technol-
ogy in augmenting the welfare of cultured animals, particularly in
terms of immunity.
Thus, in order to assess the viability of this novel technology for
indoor culture of rohu, the experiments were designed (1) to
elucidate the water quality dynamics, particularly the mechanisms
inuencing the inorganic nitrogen removal, (2) to investigate the
impact of such a system on the zootechnical performances such as
growth and survival of the cultured species and (3) to assess the
immune responses of the cultured sh.
2. Materials and methods
2.1. Experimental design and sh stocking
The experiment was carried out in indoor breglass reinforced
plastic (FRP) tanks with a water volume of 500 L each as described
in our previous study (Kheti et al., 2017). The biooc group con-
sisted of three tanks with molasses addition and the control group
(three tanks) was without the addition of molasses. The tanks were
allocated following a completely randomized design. The tanks
were lled with dechlorinated water and continuous aeration was
provided in each tank using a 90 W air blower (VB-185G; output
300 L min1; Hailea, China) connected to diffuser air stones to meet
the oxygen demand. Before sh stocking, each tank was added with
250 ml of concentrated biooc developed in a separate indoor tank
using aquaculture pond bottom soil as inoculum followed by
addition of ammonium sulphate as initial source of nitrogen,
molasses as carbon source and some amount of broken feed.
Fingerlings of mixed sex rohu (Labeo rohita) with a mean weight
9
of 7.43 0.54 g were purchased from a local sh farm and accli-
matized for two weeks. After acclimatization, sh were randomly
distributed in each tank (20 sh tank1). The sh in biooc group
were cultured with daily addition of molasses as carbon source to
maintain the C/N ratio at 15. The amount of molasses to be added
was determined following the method as described previously
(Crab et al., 2012). During the experimental period, water was not
exchanged in the biooc tanks, except the addition of dechlorinated
water to compensate evaporation loss. The sh in biooc and
control tanks were fed daily with a pelleted feed prepared from
locally available ingredients (crude protein: 22.39 0.72%, mois-
ture: 6.84 0.22%, crude lipid: 7.30 0.41%, ash: 12.21 0.68%,
crude bre: 13.87 0.49% and nitrogen free extract: 37.39 1.14%)
at 3% of body weight. The experiment was carried out for a period of
16 weeks.
2.2. Assessment of water quality parameters
During the experimental period, water temperature, dissolved
oxygen (DO; YSI Pro), pH (HANNA, USA) were determined at
weekly interval. Water samples (100 mL) were collected from each
tank and ltered through pre-dried and pre-weighed GF/C lter
paper. Total ammonia nitrogen (TAN), nitrite (NO2eN) and nitrate
(NO3eN) concentrations were measured spectrophotometrically
(Multiskan GO, Thermoscientic, USA) from the ltered water
following standard methods (APHA, 1998). Filter paper was used for
determination of total suspended solids (TSS).
2.3. Determination of proximate composition of biooc
The biooc materials were collected from individual tanks at
weekly interval. The agitation of water in the tanks was stopped by
closing the air blower for 1 h and 2 L oc water was collected from
the bottom of each tank. The oc water was then passed through
nylon lter of 10 mm mesh size and aliquots of the collected oc
were ground and processed for proximate analysis using standard
methodologies (AOAC, 1990).
2.4. Assessment of survivability, growth and sh welfare indicators
At the end of experimental period, sh were harvested from
each tank and nal body weights were measured. Survival rate,
weight gain (WG) and weight gain per day (WGD) were calculated
using the following equations: Survival rate (%) 100  (nal sh
count initial sh count), WG (%) 100  (nal body weight -
initial body weight) initial body weight, WGD (nal body
weight - initial body weight) total days of culture.
Sampling of sh and collection of blood and serum were per-
formed as described previously (Kheti et al., 2017). Briey, ve sh
from each tank were randomly sampled, euthanized (using clove
oil @ 100 mL L1) and bled with the help of a sterilized 1 mL hy-
podermal syringe from caudal vein. Pooled blood from ve sh of
each tank was allowed to clot at room temperature for 30 min,
centrifuged at 5000 g for 5 min and the sera thus obtained, were
stored at 20  C in sterilized vial until further use. For nitroblue
tetrazolium (NBT) assay, aliquot of blood was kept separately with
EDTA as anticoagulant. The NBT assay was performed from blood
phagocytes as described by Anderson and Siwicki (1995). The total
serum myeloperoxidase content was determined as described by
Mohanty and Sahoo (2010). The total serum protein content was
estimated by the biuret method (Gornall et al., 1949) and the
alkaline phosphatise activity was measured following the method
of Kind and King (1954). Finally, to measure the stress, glucose
content of serum was estimated as described by Triender (1969).
10 D. Kamilya et al. / Chemosphere 182 (2017) 8e14

Table 1
Water quality parameters in biooc and control tanks.

Water quality variables Treatments

Control Biooc

a
Temperature ( C) 27.06 1.31 (24e29) 27.06 1.31 (24e29)a
Dissolved Oxygen (mg L1) 6.53 0.77 (4.6e8.8)a 5.4 0.56 (4.4e7.0)b
pH* 6.59, 7.1a 7.07, 7.28 b
Total ammonia nitrogen (mg L1) 0.06 0.05 (0.01e0.17)a 0.11 0.05 (0.03e0.24)b
Nitrite (mg L1) 0.03 0.02 (0.00e0.09)a 0.03 0.02 (0.01e0.10)b
Nitrate (mg L1) 0.06 0.02 (0.02e0.12)a 0.03 0.02 (0.01e0.09)b
Total suspended solids (mg L1) 23.85 19.43 (1.20e70.00)a 284.48 264.17 (23.40e1328.40)b

Different superscript letters within a row indicate statistically signicant differences (P < 0.05) between treatments. Data are presented as mean SD (range).
*
Values indicate the 95% condence interval for mean pH.

2.5. Statistical analysis
Data were analyzed using multivariate statistics as exploratory
methods. Factor analysis was performed to identify the water
quality parameters that account for the main variability among the
measured variables to elucidate the mechanism of inorganic ni-
trogen removal. A correlation matrix was generated for all the
water quality variables. Then, factors were extracted from the
correlation matrix using principal component analysis and nally
varimax rotation with Kaiser Normalization was followed. A factor
loading matrix representing the variables and extracted factors was
generated. Within the matrix, a higher factor loading (represented
by the coefcients of the linear functions) indicates a dominance of
the corresponding variable. Factor loading with the same algebraic
sign implies a direct relationship between the variables and the
factor. Conversely, an inverse relationship exists if they are repre-
sented with opposite signs.
Different water quality parameters were compared between
biooc and control system using two-way ANOVA. Growth, stress
and immunological parameters were compared between the
treatments using one-way ANOVA. Results are presented as
mean standard deviation (SD). Probability levels of 0.05 were
used to nd out the signicance in all cases. All the analysis of data
was performed using SPSS-16.0 for windows software (SPSS Inc.,
Chicago, IL, USA).
3. Results and discussion
3.1. Water quality parameters
In the biooc system, water temperature (27.06 1.31  C), DO
concentrations (5.4 0.56 mg L1) and pH (7.07, 7.28; 95% con-
dence interval for mean) were well within the acceptable limits
required for tropical aquaculture (Bhatnagar and Devi, 2013). The
values of temperature (27.06 1.31  C), DO (6.53 0.77 mg L1)
and pH (6.59, 7.1; 95% condence interval for mean) were also
within the normal range in control system. Although same amount
of aeration was provided to each tank, the DO levels were signi-
cantly lower (P < 0.05) in the biooc system than in the control
treatment (Table 1). This may be due to the higher oxygen utiliza-
tion during nitrication coupled with nitrogen immobilization
through heterotrophic pathway of microbial production which is
common in biooc system (Sharma and Ahlert, 1977; Azim et al.,
2008; Ray et al., 2009). A decrease in pH value is generally

Biofloc
Control
9.00

8.00

7.00
pH

6.00

5.00

4.00
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Sampling Week

Fig. 1. The pH in control and biooc tanks throughout the experimental period. Data are represented as mean SD of three replications per sampling week.
 D. Kamilya et al. / Chemosphere 182 (2017) 8e14
A of NO2eN and NO3eN may be due to the dominance of hetero-
B Gradual increase coupled with high degree of uctuation (high
C Factor I accounted for 40.09% of the overall data variability and
Fig. 2. Different inorganic nitrogen species in control and biooc tanks throughout the
experimental period. A: Total ammonia nitrogen, B: Nitrite, C: Nitrate. Data are rep-
resented as mean SD of three replications per sampling week.
associated when there is nitrication in biooc based system (Azim
et al., 2008; Azim and Little, 2008; da Silva et al., 2013). However,
the pH remained almost stable through the experimental period in
the present study except lower values in control treatment at the
end of the culture period (Fig. 1).
Different inorganic nitrogen species (TAN, NO2eN and NO3eN)
had signicant variability in relation with that observed in control
treatment. The concentrations of all the inorganic nitrogen species
showed wide uctuation during the entire experimental period
(Fig. 2). While the TAN concentrations were always higher in biooc
system than the control (Fig. 2A), the reverse trend was observed in
case of NO2eN (Fig. 2B) and NO3eN (Fig. 2C). Unlike TAN, the
NO2eN and NO3eN concentrations were signicantly lower
11
(P > 0.05) than the control (Table 1). The generation of lower levels
trophic pathway of ammonia immobilization. In fact, biooc system
with proper C/N ratio encourages immobilization of ammonia by
heterotrophic bacteria rather than nitrication (Avnimelech et al.,
1989; Burford et al., 2004). In the biooc system, the NO3eN con-
centration decreased gradually after 7th week indicating a lesser
degree of nitrication. In contrary, the NO3eN concentration in
control tanks decreased during 5e7th weeks followed by an
increasing trend until the end of the experimental period indicating
a better nitrication. Overall, the dynamics of three inorganic ni-
trogen species in the biooc system indicated involvement of
ammonia immobilization through heterotrophic pathway as well
as nitrication.
The TSS levels in the biooc and control systems were
284.48 264.17 mg L1 and 23.85 19.43 mg L1, respectively.
standard deviation) was observed in the TSS levels throughout the
experimental period (Table 1, Fig. 3). Even though microbial ocs
were periodically removed from the tanks, their level became too
high during last few sampling weeks. The highest TSS level recor-
ded from biooc treatment was 1328.40 mg L1 during 14th week
whereas it was 70.0 mg L1 in control tank during 7th week. Similar
observations have also been reported from indoor biooc system
with cultured sh (Azim and Little, 2008).
The results of the factor analysis in the form of factor loading
matrix are presented in Table 2. In this study, two factors were
drawn out of the seven variables that accounted for 58.44% of the
overall data variability. The rst factor extracted from the correla-
tion matrix was the linear combination of the original variables
accounting for the highest variation contained in the samples. The
second factor accounted for most of the remaining variability.
had a high positive loading in DO and NO3eN. On the other hand,
temperature and TAN had high negative loading. The opposite
loading pattern of DO and temperature indicated the inverse rela-
tionship between these two parameters. The negative loading of
TAN indicated the disproportion between this parameter and pa-
rameters with high positive loading. Removal of TAN from an
aquaculture system occurs through three major pathways: (1)
photoautotrophic uptake by algae and phytoplankton, (2) immo-
bilization by heterotrophic bacteria and (3) chemoautotrophic up-
take by nitrifying bacteria (Hargreaves, 2006; Ebeling et al., 2006).
In a light limited indoor biooc system, the second and third
pathways are predominant. In the present experiment, after the
initial spike up to 5th week, the TAN concentration remained
almost same along nal weeks of the culture period (Fig. 2A). This,
coupled with the decreasing nitrate levels from 5th week up to the
end of the experiment (Fig. 2C) indicated the dominance of the
second pathway of nitrogen removal from the system. As the TAN
was subjected to heterotrophic bacterial activity, less amount was
available for nitrication as indicated by decreasing level of nitrate.
Hence, Factor 1 represents removal of TAN by heterotrophic bac-
teria mediated immobilization along with nitrication. Previous
studies reported high nitrate accumulation in biooc based culture
system indicating a dominance of nitrication (Azim and Little,
2008; Azim et al., 2008). However, in a recent study, Xu et al.
(2016) demonstrated reduced nitrate level in biooc system with
C/N ratio of 15 and 18 and inferred that the heterotrophic bacterial
production as the major pathway of ammonia removal.
Factor 2 accounted for a further 18.35% of the overall data
variability and had a high positive loading in TSS and pH and a high
negative loading in NO3eN. The positive loading of TSS indicated
the heterotrophic bacterial production under aerobic condition in
the biooc system. As the changes of TSS level over time reects the
12 D. Kamilya et al. / Chemosphere 182 (2017) 8e14

Control
Biofloc
1200

900

600
TSS (mg L-1)

300

0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Sampling Week

Fig. 3. Total suspended solids in control and biooc tanks throughout the experimental period. Data are represented as mean SD of three replications per sampling week.

Table 2 the analysis of Factor 1, it can further be conrmed that the

Results of factor analysis for water quality variables. immobilization of ammonia by heterotrophic bacteria and subse-
Water quality variables Factor 1 Factor 2 quent production of microbial biomass was the dominant pathway
Temperature 0.816 0.365
observed in the present study. Hence, Factor 2 represents hetero-
Nitrate 0.632 0.528 trophic bacterial biomass production. Generally, a decrease in pH is
Dissolved Oxygen 0.624 0.244 observed with an increase in bacterial production in biooc system
Nitrite 0.600 0.130 (Azim et al., 2008). In the present study, the pH of the biooc tanks
Total ammonia nitrogen 0.531 0.295
remained almost stable throughout the experimental period.
pH 0.019 0.884
Total suspended solids 0.472 0.645 Although an inverse relationship between TSS concentration and
% variation 40.09 18.35 pH is generally observed associated with higher nitrication ac-
Factor loadings in bold were used for interpretation.
tivity in higher TSS concentrations, the high positive loading of pH
indicated a higher respiration rate due to heterotrophic activity
associated with the molasses input and to TSS removal.
Table 3
Welfare indicators of Labeo rohita cultured in biooc and control systems. 3.2. Proximate analysis of biooc
Welfare indicators Treatments
The proximate analysis (8.57 0.26% moisture, 14.37 0.40%
Control Biooc
crude protein, 3.13 0.32% crude lipid, 40.55 0.77% ash,
Nitroblue tetrazolium activity (OD595) 0.47 0.19a 0.65 0.17b 4.52 0.03% crude bre and 28.98 0.16% nitrogen free extract on
Serum myeloperoxidase content (OD450) 1.22 0.16a 1.40 0.25a
dry matter basis) of the microbial oc revealed high ash content
Total serum protein (g dL1) 15.59 0.69a 16.37 0.29a
Alkaline phosphatase activity (IU L1) 97.44 14.38a 209.05 12.45b combined with low crude protein content. Presence of high amount
Serum glucose content (mg dL1) 201.99 9.72a 196.78 4.85a of ash in biooc has also been reported from other studies as well
Different letters indicate statistically signicant difference (P < 0.05) between (Ju et al., 2008; Bauer et al., 2012; Hende et al., 2014; Neto et al.,
treatments. 2015). Even though the nutritional quality of the biooc cant be
Data are presented as mean SD. considered as good enough due to high ash and low protein con-
tent, it can still serve as in situ derived natural food for the cultured
animals (Ray et al., 2017). However, more experimentation is
development of the biooc, it is often used as an indicator for required for evaluating the potential of oc material either as in situ
quantitative determination of microbial oc biomass (De Schryver derived natural food source or their use as ingredients in formu-
et al., 2008). The microbial biomass synthesis increased gradually lated diets.
in the biooc tanks owing to the continued fresh feed input, regular
addition of molasses and presence of optimum water quality pa-
3.3. Survivability, growth and sh welfare indicators
rameters. This microbial biomass was probably of heterotrophic
bacterial population as the microbial biomass yield per unit sub-
There was no statistically signicant difference (P > 0.05) in
strate of heterotrophic bacteria is far greater than that of nitrifying
survivability of rohu between biooc system (95.0%) and the con-
bacteria in biooc system with proper C/N condition (Ebeling et al.,
trol (93.33%). However, the weight gain of rohu cultured in biooc
2006; Hargreaves, 2006). This was also evident from high negative
system (105%) was signicantly higher (P < 0.05) than rohu
loading of NO3eN indicating reduced nitrication. Together with
cultured in control system (65.8%). Similarly, weight gain per day
 D. Kamilya et al. / Chemosphere 182 (2017) 8e14
was also signicantly higher (P < 0.05) in biooc system (1.40 g d1)
than the control (0.68 g d1). The high survivability and growth of
rohu indicated the signicant contribution of the biooc system in
enhancing these zootechnical parameters. Enhancement of such
response variables was also observed in earlier studies (Long et al.,
2015; Ahmad et al., 2016; Perez-Fuentes et al., 2016).
The serum glucose content, as sh stress indicator, did not show
signicant difference (P > 0.05) between the sh cultured in biooc
and control system (Table 3). Though not fully conclusive, it indi-
cated that the biooc system probably did not exert any stress to
the sh. In a recent study, Ahmad et al. (2016) did not observe any
stress on rohu grown in biooc system. Increased stress due to the
presence of biooc was also not observed in Nile tilapia (Azim and
Little, 2008).
NBT activity (respiratory burst activity) by phagocytes is a
widely used indicator of immunocompetence in sh (Secombes,
1990). In the present study, NBT activity showed signicant
enhancement (P < 0.05) in rohu grown in biooc system than the
control (Table 3). In a recent study, signicant enhancement of this
activity was observed in the same species cultured in biooc system
(Ahmad et al., 2016). Increased respiratory burst activity has also
been reported in genetically improved farmed tilapia (Oreochromis
niloticus) grown in biooc system for 8 weeks (Long et al., 2015).
Alkaline phosphatase is a metalloenzyme that takes part in im-
mune functions of animals (Dong et al., 2015). The results of the
present study showed signicant alkaline phosphatase activity by
rohu grown in biooc system. However, no reports are available
regarding this activity to corroborate our result.
Myeloperoxidase enzyme is released by the granules of neu-
trophils and participates in defense against pathogens (Dalmo et al.,
1997). Serum myeloperoxidase activity of rohu cultured in biooc
system did not differ signicantly (P > 0.05, Table 3) than the
control. This is in contrary to a previous study where signicantly
elevated serum myeloperoxidase level was observed in biooc
raised rohu (Ahmad et al., 2016). This discrepancy might be due to
the use of different carbon sources that favoured development of
specic group of bacteria and their specic mode of action (Kheti
et al., 2017). Like the myeloperoxidase activity, the total serum
protein content of rohu also did not differ signicantly (P > 0.05,
Table 3) than the control sh.
Overall, the welfare of rohu, as indicated by the stress and
immunological parameters, was augmented when cultured in the
biooc system. In fact, the cellular components, metabolites and
other derivatives of the heterotrophic bacterial assemblage present
in the biooc system are expected to have immunostimulatory
potential, thereby can contribute to a healthy status of cultured sh
(Crab et al., 2012).
4. Conclusions
Factor analysis revealed ammonia immobilization by hetero-
trophic bacterial population, rather than nitrication, as the
dominant mechanism inuencing the inorganic nitrogen removal
from the biooc system with cultured animals. Consequent bacte-
rial biomass production in the biooc system, as also revealed
through factor analysis, might served as an in situ derived natural
food, thereby contributed to the enhanced growth of sh. An
indication of immunity enhancement of the cultured sh was also
observed in the biooc system. However, more studies are required
to conclusively elucidate the impact of biooc system on immunity
of cultured sh.
Acknowledgment
The study was supported by a research project under OYS
13
scheme (Project code: SB/FT/LS-03/2013) funded by Science and
Engineering Research Board (SERB), New Delhi, India.
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