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WJAA #788898, VOL 00, ISS 00

Water Quality and Growth of Rohu, Labeo

Rohita, in a Biofloc System

SUDHANSU SHEKHAR MAHANAND, SANJIB MOULICK,

and P. SRINIVASA RAO

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Water Quality and Growth of Rohu, Labeo Rohita, in a Biofloc

System
Sudhansu Shekhar Mahanand, Sanjib Moulick, and P. Srinivasa Rao
Journal of Applied Aquaculture, 00:1–11, 2013
Copyright © Taylor & Francis Group, LLC
ISSN: 1045-4438 print/1545-0805 online
DOI: 10.1080/10454438.2013.788898

Water Quality and Growth of Rohu, Labeo

Rohita, in a Biofloc System

SUDHANSU SHEKHAR MAHANAND1 , SANJIB MOULICK2 ,

and P. SRINIVASA RAO1
1
Agricultural and Food Engineering Department, Indian Institute of Technology, Kharagpur, 5
West Bengal, India
2
School of Civil Engineering, KIIT University, Bhubaneswar, Odisha, India

We evaluated the technical feasibility of reducing water

dependency of rohu, Labeo rohita, culture with biofloc under
light-limited indoor culture. Biofloc and control treatments were 10
conducted in 700-L indoor tanks at three different stocking densi-
ties (STD): 1.3, 2.6, and 3.9 Nos. fish m−2 of surface area of tank
for a period of 90 days. In biofloc treatment, fish were fed 20%
crude protein feed and extra organic carbon in the form of wheat
flour, whereas in case of control treatment they were fed 30% crude 15
protein feed only. Fish survival was 100% in both the treatments.
Lower stocking density produced larger fish, but growth was simi-
lar within stocking densities among control and biofloc treatments.
The nutritional quality of biofloc was found to be quite suitable for
rohu. Frequency of water exchange was significantly less in biofloc 20
treatments as compared to the control ones.

KEYWORDS Biofloc technology, Rohu, inorganic nitrogen

removal, water quality

INTRODUCTION

Among the Indian major carps, rohu (Labeo rohita) is preferred, representing 25
about 35% of production (Food and Agriculture Organization of the
United Nations 2000). New technology has raised production levels to 3-
5 tons/ha/yr in the best cases, but with high water demand. Controlling

Address correspondence to Sanjib Moulick, Associate Professor (II), School

of Civil Engineering, KIIT University, Bhubaneswar, 751024, Odisha, India. E-mail:
sanjib_moulick72@yahoo.co.uk

1
2 S. S. Mahanand et al.

nitrogenous wastes in more intensive systems requires frequent water

exchange, leading to environmental degradation and high pumping costs 30
(Avnimelech 1999). Two relatively new technological approaches to reducing
water consumption in aquaculture are: 1) recirculating aquaculture systems
(RAS) and 2) biofloc. Schryver et al. (2008) compared operating costs of a
nitrifying trickling filter and biofloc technology for tilapia culture and found
that the biofloc system was about 50% cheaper than biofiltration. 35
In biofloc systems, nitrogenous waste is reduced through micro-
bial assimilation aided by the addition of extra carbonaceous materials
(Avnimelech 1999). The heterotrophic microbial biomass is suspected to have
a controlling effect on pathogenic bacteria (Michaud et al. 2006). Several
studies have been conducted using biofloc technology for culture of tilapia 40
(Azim and Little 2008; Asaduzzaman et al. 2009) and shrimps (Kuhn et al.
2008; Kuhn et al. 2009; Kuhn et al. 2010; Asaduzzaman 2010; Ray et al. 2011).
However, no study has been reported on the growth and production of rohu
using biofloc technology. This study explores water use in a light-limited
indoor biofloc system stocked with rohu, Labeo rohita, at three densities. 45

MATERIALS AND METHODS

Experiments were carried out over 90 days at the Indian Institute of

Technology-Kharagpur in 18 cement circular tanks of diameter 1.06 m and
height of 0.91 m, with a water volume kept at 700 L. Mixed sex rohu (Labeo
rohita) of individual average weight 50 ± 1.5 g were grown out in triplicated 50
biofloc and control systems at three different stocking densities: 1.3, 2.6, and
3.9 fish m−2 of tank surface area. The surface area of the tank was 0.88 m2 .
In the biofloc system, fish were fed 20% crude protein pellets plus extra
organic carbon in the form of wheat flour to maintain a C:N ratio in the
inputs (feed + wheat flour) of 10:1 (Avnimelech 1999). In the control, fish 55
were fed 30% crude protein feed only. Percentage of raw ingredients and
proximate composition, energy content, and C:N ratios of the experimental
diets for both the treatments are given in Table 1. Feeding rates were based
on observation of feeding behavior of fish during the first several days, fixed
at 2.0% of the total stocked biomass daily, and adjusted fortnightly after 60
sampling. Daily feed rations were split into two equal amounts given at
09:00 and 18:00 h to all tanks.
The amount of wheat flour required daily for the biofloc treatments
was calculated following Schryver et al. (2008) as 15 xypk grams dry matter
per m3 water; where, x = fish density (g/m3 of water volume), y = feed rate 65
as a proportion of body weight of fish (e.g., 2.0% = 0.02), p = proportion of
protein in feed, and k = proportion of nitrogen in feed. It was assumed that
75% of the total feed-N ends up in water (Piedrahita 2003) and that wheat
flour contains 50% organic carbon. In the present study, y, p, and k were
 Rohu Culture Using Biofloc Technology 3
Q3

TABLE 1 Feed ingredients and biochemical composition of the experimental diet.

High protein diet Low protein diet

(a) Feed ingredients (%)

Fishmeal 26 14
Groundnut oil–cake 25 25
Rice bran 38 50
Wheat floor 10 10
Vitamin and mineral mix 1 1
(b) Biochemical composition (%DM)
Moisture 6.63 4.5
Crude protein 30.50 20.58
Crude lipid 3.50 3.55
Ash 19.38 18.12
Crude fiber 13.30 13
Energy (kJg−1 ) 11.62 11.1
Nitrogen-free extract 27.06 40.25
C:N ratio 10.6 16

2.0%, 20%, and 16%, respectively. C:N ratios were determined using a CHN 70
analyzer (Perkin Elmer 2400 Series II).
Tanks were aerated and agitated continuously using air stones. Water
temperature, dissolved oxygen (DO), and pH were determined daily using a
HACH Hydrolab DS5. DO concentration averaged 7.5 mg/l with a range
of 6.45–8.68 mg/l, and pH averaged 8.07, with a range of 6.94–8.65. 75
In the biofloc system, NaHCO3 was added on several occasions when pH
declined below 7.0. Water temperature averaged 27.5◦ C, (range 21◦ –31.7◦ C).
Nitrite-nitrogen (NO2 –N), nitrate-nitrogen (NO3 –N), and total ammonia nitro-
gen (TAN) were analyzed using a spectrophotometer (UV 1700 Shimadzu
Spectrophotometer) according to standard methods (APHA 2005). Total sus- 80
pended solid (TSS) and biochemical oxygen demand (BOD5 ) of water was
measured weekly (around 12:00 noon) following the methods of Stirling
(1985). In all the treatments whenever TAN approached 1 mg/L, 50% of the
tank water was exchanged.
Water samples were collected fortnightly from biofloc tanks for bio- 85
chemical analysis. Concentrated floc samples were collected from each tank
using a muslin cloth, dried in an oven at 102◦ C to constant weight, and
preserved in a refrigerator. At the end of the experiment, tank-wise pooled
samples were ground and processed for proximate analysis following AOAC
methods (1990). Fatty acid composition was determined in samples extracted 90
for total lipids, according to Bligh and Dyer (1959) as modified by Christee
(1982), Mangold (1969), and Ackman and Burgher (1965).
More than 50% of fish in each tank were sampled fortnightly and
measured individually to estimate (1) specific growth rate (SGR), (2) feed
conversion ratio (FCR), (3) protein efficiency ratio (PER), and (4) net 95
4 S. S. Mahanand et al.

fish yield (NFY). These parameters are calculated based on the following
equations:

SGR (% body weight/day) = 100 × ln (final weight/initial weight)/

culture period (days)

FCR = amount of feed fed (dry weight basis)/net weight gain

(wet weight basis)

PER = net weight gain (wet weight basis)/amount of protein dosed

(dry weight basis)

NFY (kg/ha) = [total biomass at harvest (kg) − total biomass at stocking

(kg)] × 104 /surface area of the culture tank (m2 )

Data resulting from the experiments were subjected to one-way analysis

of variance. Tukey’s honestly significant difference (HSD) test was used to
identify differences among treatments at a significance level of 5%. 100

RESULTS AND DISCUSSION

TSS and BOD5 data are presented in Table 2. Both tended to increase with
stocking density and time. Overall, the biofloc system had much higher TSS
and BOD5 . At the last sampling, maximum TSS and BOD5 were recorded at
the last sampling (day 90) as 204 mg/L and 106 mg/L, respectively, in the 105
biofloc tank stocked with 3.9 fish/m2 of surface area. This compared well
with work reported by Azim and Little (2008) for indoor biofloc tanks stocked
with Nile tilapia (Oreochromis niloticus) where maximum TSS and BOD5
reached 1,000 and 290 mg/L, respectively, without any water exchange.
Periodic water exchange (whenever TAN approached 1 mg/l) undoubtedly 110
lowered TSS and BOD in our study.
Average dissolved inorganic nitrogen (TAN, NO2 , and NO3 ) concentra-
tions over the experimental period are shown in Figure 1. Sudden declines
denote water exchanges made to maintain TAN below 1 mg/L. TAN con-
centration tended to increase linearly between water exchanges (Figure 1a). 115

TABLE 2 Mean (±SD) TSS and BOD values in control and biofloc treatments.

Control Biofloc
2 2 2 2
Parameter 1.3 no/m 2.6 no/m 3.9 no/m 1.3 no/m 2.6 no/m2 3.9 no/m2

TSS (mg/l) 10.5 ± 6.8 12.2 ± 8.3 13.5 ± 9.3 66.5 ± 46 80 ± 56.7 92.9 ± 68.5
BOD5 (mg/l) 6.3 ± 3.4 7.5 ± 4.2 9.2 ± 5.2 29.7 ± 19.1 41.8 ± 26.3 50.7 ± 33
 Rohu Culture Using Biofloc Technology 5

(A) 1.2

0.8
TAN, mg/l
0.6

0.4

0.2 STD 1.3 no/m2_Control tank STD 2.6 no/m2_Control tank

STD 3.9 no/m2_Control tank STD 1.3 no/m2_Biofloc tank
STD 2.6 no/m2_Biofloc tank STD 3.9 no/m2_Biofloc tank
0
25-Aug-10

1-Sep-10

8-Sep-10

15-Sep-10

22-Sep-10

29-Sep-10

6-Oct-10

13-Oct-10

20-Oct-10

27-Oct-10

3-Nov-10

10-Nov-10

17-Nov-10
Time, date
(B) 0.5
STD 1.3 no/m2_Control tank STD 2.6 no/m2_Control tank
STD 3.9 no/m2_Control tank STD 1.3 no/m2_Biofloc tank
0.45 STD 2.6 no/m2_Biofloc tank STD 3.9 no/m2_Biofloc tank
0.4
0.35
Nitrite-N, mg/l

0.3
0.25
0.2
0.15
0.1
0.05
0
25-Aug-10

1-Sep-10

8-Sep-10

15-Sep-10

22-Sep-10

29-Sep-10

6-Oct-10

13-Oct-10

20-Oct-10

27-Oct-10

3-Nov-10

10-Nov-10

17-Nov-10

Time, date
(C) 40 2
STD 1.3 no/m _Control tank STD 2.6 no/m2_Control tank
STD 3.9 no/m2_Control tank STD 1.3 no/m2_Biofloc tank
35
STD 2.6 no/m2_Biofloc tank STD 3.9 no/m2_Biofloc tank
30
Nitrate-N, mg/l

25

20

15

10

0
25-Aug-10

1-Sep-10

8-Sep-10

15-Sep-10

22-Sep-10

29-Sep-10

6-Oct-10

13-Oct-10

20-Oct-10

27-Oct-10

3-Nov-10

10-Nov-10

17-Nov-10

Time, date

FIGURE 1 Dissolved inorganic nitrogen in different treatments throughout the experimental

period: (A) TAN, (B) NO2 -N, and (C) NO3 -N. Values are means of three replications in each
alternate date for biofloc and control treatments, respectively. The standard deviations were
within ±5% (not shown in the figures). (Color figure available online.)
6 S. S. Mahanand et al.

Nitrite and nitrate concentrations increased with time, with only slight reduc-
tions associated with water exchange (Figure 1b, 1c). Biofloc tanks had
generally lower inorganic N concentrations than control tanks. Higher stock-
ing densities also led to higher inorganic N. In biofloc treatments, higher
BOD5 (29.7 to 50.7 mg/L) reduced inorganic nitrogenous compounds as 120
these were assimilated by heterotrophic biomass (Table 2).
The number of water exchanges conducted in the different treatments
and total consumptive water use (L/kg) are presented in Table 3. The total
consumptive use of water is significantly different in two different treatments
with different stocking densities. The needed number of water exchanges 125
was more in treatments with higher stocking densities and less in biofloc
treatments.
Proximate analysis of biofloc is presented in Table 4. Biofloc contained
35% protein, 1% lipid, 15% fiber, 15% ash, and 19 kJ/g energy on a dry
matter basis. About 30% crude protein is sufficient for optimum growth of 130
rohu (Singh et al. 2006). The biofloc that developed in our study is adequate
for rohu culture except for somewhat high fiber and ash, which decreases
the quantity of the usable nutrient in the diet (De Silva and Anderson 1995).
The biofloc was further analyzed to see whether it contained essential fatty
acids for the fish (Table 5). There were 21.11% polyunsaturated, 35.63% 135

TABLE 3 Details of water exchange in control and biofloc treatments.

Number of
Stocking water Total Consumptive Days of water
Density (m−2 ) Treatments exchanges Water Use (L/kg) exchange

1.3 Control 4 345 30,42,60,72

Biofloc 3 208 53,65,86
2.6 Control 4 208.5 23,50,65,83
Biofloc 3 140.4 31,62,77
3.9 Control 5 200.5 20,32,47,62,77
Biofloc 4 133.7 44,59,71,83

TABLE 4 Mean (±SD) proximate composition,

energy content, and C:N ratio of biofloc.

Composition (% DM) Values

Moisture 4.1 ± 0.10

Crude protein 35.40 ± 0.63
Crude lipid 1.1 ± 0.26
Ash 15.38 ± 0.35
Crude fibre 15.03 ± 0.37
Energy (kJg−1 ) 18.78 ± 0.19
C:N ratio 7.0 ± 0.48
 Rohu Culture Using Biofloc Technology 7

TABLE 5 Fatty acid composition (% lipid) of

biofloc.

Componentsa Values

12:0 0.3
13:0 0.2
13:1 0.3
14:0 2.9
14:1 4.5
15:0 2.3
15:1 0.9
16:0 29.3
16:1 7.9
16:2 2.0
17:0 0.2
17:1 0.3
18:0 4.6
18:1ω9 20.8
18:2ω6 12.7
18:3ω6 0.6
18:3ω3 4.4
20:0 0.2
20:1ω9 0.5
20:3ω6 0.3
20:4ω6 0.1
22:0 1.9
20:4ω3 −
22:1ω11 0.4
20:5ω3 0.4
21:5ω3 0.3
22:5ω6 0.01
24:0 1.2
24:1 0.03
22:6ω3 0.3
Total -ω3 5.4
Total -ω6 13.71
Total PUFA 21.11
Total saturates 43.1
Total monoenes 35.63
Total lipid % 4.22
a
First and second figures represent carbon chain length
and number of double bonds, respectively. Q1
The ω values represent the methyl end chain from the
center of double bond furthest from the carboxyl end.

monounsaturated, and 43.1% saturated fatty acids in the biofloc sample.

Tacon (1990) reported that about 1% of 18:2 ω6 and 18:3 ω3 essential fatty
acids are required for common carp. In the biofloc samples those values
were 12% and 4.4%.
Fish growth data are presented in Table 6. Fish survival was 100% in 140
both treatments. The biofloc treatment generated greater individual weight
gain and net fish production than the corresponding control treatments. Net
 TABLE 6 Fish growth parameters of rohu in both control and biofloc treatments.

Treatments

Control Biofloc

STD (no/m2 ) 1.3 2.6 3.9 1.3 2.6 3.9

Initial individual weight (g) 56.75 ± 2.53a 58.41 ± 0.85b 54.86 ± 4.00c 53.33 ± 8.03a 52.5 ± 3.30b 48.61 ± 2.40c
Final individual weight (g) 112.75 ± 3.19a 93.29 ± 2.46b 80.86 ± 1.14c 140 ± 16.39a 103.95 ± 1.30b 97.36 ± 3.75c

8
Individual weight gain (g) 56 ± 0.66a 34.87 ± 1.75b 26 ± 2.88c 86.66 ± 8.77a 51.45 ± 4.16b 48.75 ± 6.00c
Survival (%) 100 100 100 100 100 100
NFY (kg/ 700 L) 4.5 ± 0.01 7.5 ± 0.02 9.7 ± 0.01 5.6 ± 0.04 8.3 ± 0.01 11.7 ± 0.04
SGR (%body weight day−1 ) 0.76 ± 0.018 0.52 ± 0.017 0.43 ± 0.065 1.07 ± 0.014 0.76 ± 0.08 0.77 ± 0.028
FCR 2.54 ± 0.30a 3.76 ± 0.31b 4.58 ± 0.55c 1.93 ± 0.33a 3.55 ± 0.49b 3.66 ± 0.69c
PER 1.96 ± 0.28a 1.32 ± 0.21b 1.08 ± 0.15c 2.58 ± 0.35a 1.28 ± 0.18b 1.36 ± 0.24c

Data shown are mean (±SD) for 90 days culture period. In each row, similar superscript letters indicate significant difference at 0.05 level in similar stocking density.
 Rohu Culture Using Biofloc Technology 9

yield (kg ha−1 90 day−1 ) at harvest for biofloc treatments with 1.3, 2.6, and
3.9 fish per m2 surface area was 54.8%, 47.5%, and 87.5% more than the
corresponding control treatments. Net fish yield was highest (1912 ± 16.4 kg 145
ha−1 90 day−1 ) in the biofloc tanks with the highest stocking density (3.9), but
the biofloc treatment with 1.3 fish/m2 had the lowest FCR (1.93 ± 0.33) and
highest SGR (1.07 ± 0.014) and PER (2.58 ± 0.35).
This study demonstrates the potential of biofloc technology in the
culture of rohu, Labeo rohita. The nutritional quality of biofloc was found 150
to be suitable for rohu. The highest net yield obtained in the biofloc
treatment was almost 87% more than that in corresponding control treat-
ment. Nevertheless, frequent water exchange was required to maintain toxic
ammonia-nitrogen concentrations below a lethal level, though requirement
of water exchange was less in biofloc tanks. Further study is required on 155
biofloc technology to make it self-sustaining.

REFERENCES

Ackman, R. G., and R. D. Burgher. 1965. Cod liver oil fatty acids as secondary
reference standards in the GLC of polyunsaturated fatty acids of animal origin:
Analysis of a dermal oil of the Atlantic leather-back turtle. Journal of American 160
Oil Chemist’s Society 42: 38–42.
Association of Official Analytical Chemists (AOAC). 1990. Official methods of
analysis, 15th ed. Gaithersburg, MD: AOAC Press.
Avnimelech, Y. 1999. Carbon and nitrogen ratio as a control element in aquaculture
systems. Aquaculture 176(3-4): 227–235. 165
Azim, M. E., M. A Wahab, A. A van Dam, M. C. M Beveridge, E. A. Huisman, and M.
C. J. Verdegem. 2001. Optimization of stocking ratios of two Indian major carps,
rohu (Labeo rohita Ham.) and catla (Catla catla Ham.) in a periphyton-based
aquaculture system. Aquaculture 203(1-2): 33–49.
Azim, M. E., M. C. J Verdegem, H. Khatoon, M. A. Wahab, A. A. van Dam, and M. C. 170
M. Beveridge. 2002.A comparison of fertilization, feeding and three periphyton
substrates for increasing fish production in freshwater pond aquaculture in
Bangladesh original research. Aquaculture 212(1–4): 227–243.
Azim, M. E., M. C. J. Verdegem, M. M. Rahman, M. A. Wahab, A. A. van Dam, and
M. C. M. Beveridge. 2002. Evaluation of polyculture of Indian major carps in 175
periphyton-based ponds. Aquaculture 213(1-4): 131–149.
Azim, M. E., M. A. Wahab, P. K. Biswas, T. Asaeda, T. Fujino, and M. C. J. Verdegem.
2004a. The effect of periphyton substrate density on production in freshwater
polyculture ponds. Aquaculture 232(1-4): 441–453.
Azim, M. E., M. M. Rahaman, M. A. Wahab, T. Asaeda, D. C. Little, and M. C. J. 180
Verdegem. 2004b. Periphyton-based pond polyculture system: A bioeconomic
comparison of on-farm and on-station trials. Aquaculture 242(1-4): 381–396.
American Public Health Association (APHA). 2005. Standard methods for the
examination of water and wastewater, 21st ed. Washington, DC: APHA.
10 S. S. Mahanand et al.

Avnimelech, Y. 2006. Bio-filters: The need for a new comprehensive approach. 185
Aquacultural Engineering 34: 172–178.
Azim, M. E., and D. C. Little. 2006. Intensifying aquaculture production through
new approaches to manipulating natural food. CAB Reviews: Perspectives in
Agriculture, Veterinary Science, Nutrition, and Natural Resources 1(62): 23.
Avnimelech, Y. 2007. Feeding with microbial flocs by tilapia in minimal discharge 190
bioflocs technology ponds. Aquaculture 264: 140–147.
Azim, M. E., and D. C. Little. 2008. The biofloc technology (BFT) in indoor tanks:
Water quality, biofloc composition, and growth and welfare of Nile tilapia
(Oreochromis niloticus). Aquaculture 283(1-4): 29–35.
Azim, M. E., D. C. Little, and J. E. Brone. 2008. Microbial protein production in 195
activated suspension tanks manipulating C: N ratio in feed and the implications
for fish culture. Bioresource Technology 99:3, 590–3, 599.
Asaduzzaman, M., M. A. M. Wahab, M. C. J. Verdegem, S. Benerjee, T. Akter, M. M.
Hasan, and M. E. Azim. 2009. Effects of addition of tilapia Oreochromis niloticus
and substrates for periphyton developments on pond ecology and production in 200
C/N-controlled freshwater prawn Macrobrachium rosenbergii farming systems.
Aquaculture 287(3-4): 371–380.
Bligh, E. G., and W. J. Dyer. 1959. A rapid method of total lipid extraction and
purification. Canadian Journal of Biochemistry and Physiology 37: 911–917.
Christie, W. W. 2003. Lipid analysis: Isolation, separation, identification and 205
structural analysis of lipids, 3rd ed. Cambridge, UK: The Oily Press.
Christie, W.W. 1982. Lipid analysis, 2nd ed. Oxford, UK: Pergamon Press.
Craig, S., and L. A. Helfrich. 2002. Understanding fish nutrition, feeds, and feeding.
Virginia Cooperative Extension Bulletin 4: 420–256.
De Silva, S. S., and T. Anderson. 1995. Fish Nutrition in aquaculture. London, UK: 210
Chapman and Hall.
Food and Agriculture Organization of the United Nations (FAO). 2000. Fishery
statistics (Aquaculture production). Rome, Italy: FAO.
Food and Agriculture Organization of the United Nations (FAO). 2004. The state of
world fisheries and aquaculture. Rome, Italy: FAO Fisheries Department. 215
Kuhn, D. D., G. D. Boardman, S. R. Craig, G. J. Flick, and E. Mclean. 2008. Use
of microbial floc generated from tilapia effluent as a nutritional supplement for
shrimp, Litopenaeus vannamei in recirculating aquaculture system. Journal of
World Aquaculture Society 39: 72–78.
Kuhn, D. D., G. D. Boardman, A. L. Lawrence, L. Marsh, and G. J. Flick Jr. 2009. 220
Microbial floc meal as a replacement ingredient for fish meal and soybean
protein in shrimp feed. Aquaculture 296(1-2): 51–57.
Kuhan, D. D., A. L. Lawrence, G. D. Boardman, S. Patnaik, L. Marsh, and G. J. Flick
Jr. 2010. Evaluation of two types of bioflocs derived from biological treatment of
fish effluent as feed ingredients for Pacific white shrimp, Litopenaeus vannamei. 225
Aquaculture 303: 28–33.
Mangold, H. K. 1969. In Thin layer chromatography, edited by E. Stahl, 155. New
York: Springer. Q2
Michaud, L., J. P. Blancheton, V. Bruni, and R. Piedrahita. 2006. Effect of partic-
ulate organic carbon on heterotrophic bacterial populations and nitrification 230
efficiency in biological filters. Aquacultural Engineering 34(3):224–233.
 Rohu Culture Using Biofloc Technology 11

Martins, C. I., M. G. Pistrin, S. S. W. Ende, E. H. Eding, and J. A. J. Verreth. 2009a.

The accumulation of substances in recirculating aquaculture systems (RAS)
affects embryonic and larval development in common carp Cyprinus carpio.
Aquaculture 291: 65–73. 235
Martins, C. I. M., D. Ochola, S. S. W. Ende, E. H. Eding, and J. A. J. Verreth. 2009b. Is
growth retardation present in Nile tilapia Oreochromis niloticus cultured in low
water exchange recirculating aquaculture systems? Aquaculture 298: 43–50.
Piedrahita, R. H. 2003. Reducing the potential environmental impact of tank
aquaculture effluents through intensification and recirculation. Aquaculture 226: 240
35–44.
Ray, A. J., K. S. Dillon, and J. M. Lotz. 2011. Water quality dynamics and shrimp
(Litopenaeus vannamei) production in intensive, mesohaline culture systems
with two levels of biofloc management. Aquacultural Engineering 45: 127–136.
Schneider, O., J. P. Blancheton, L. Varadi, E. H. Eding, and J. A. J. Verreth. 2006. 245
Cost price and production strategies in European recirculation systems: Linking
tradition & technology highest quality for the consumer. Florence, Italy: World
Aquaculture Society.
Sahu, P. K., J. K. Jena, P. C. Das, S. Mondal, and R. Das. 2007. Production per-
formance of Labeo calbasu (Hamilton) in polyculture with three Indian major 250
carps Catla catla (Hamilton), Labeo rohita (Hamilton) and Cirrhinus mrigala
(Hamilton) with provision of fertilizers, feed and periphytic substrate as varied
inputs. Aquaculture 262(2-4): 333–339.
Schryver, P. D., R. Crab, T. Defoirdt, N. Boon, and W. Verstraete. 2008. The basics of
bio-flocs technology: The added value for aquaculture. Aquaculture 277(3-4): 255
125–137.
Singh, P. K., S. R. Gaur, and M. S. Chari. 2006. Effect of varying protein levels on
the growth of Indian major carp rohu, Labeo rohita (Hamilton). International
Journal of Zoological Research 2(2): 186–191.
Stirling, H. P. 1985. Chemical and biological methods of water analysis for 260
Aquaculturists. Stirling, Scotland: Institute of Aquaculture, University of Stirling.
Tacon, A. G. J. 1990. Standard methods for the nutrition and feeding of farmed fish
and shrimp. Washington DC: Argent Laboratories Press.

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