A Seminar paper on

Biofloc technology and its potentiality for higher production of fish in

Bangladesh

Course code: GFB 598

Term: summer’20

Submitted to

Course Instructors Major Professor

Dr. A. K. M. Aminul Islam
Professor, Dept. of Genetics and Plant Dr. Zinia Rahman
Breeding Associate Professor,
Dept. of Genetics and Fish Breeding
Prof. Dr. Md. Mizanur Rahman BSMRAU, Gazipur 1706
Professor, Dept. of Soil Science

Dr. Md. Sanaullah Biswas

Associate Professor, Dept. of Horticulture

Dr. Dinesh Chandra Shaha

Associate Professor, Dept. of Fisheries
Management

BSMRAU, Gazipur 1706

Submitted By

Rabeya Chowdhury Shapla

Program- Ms
Reg. No- 15-05-3606
Dept. of Genetics and Fish Breeding

Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur

1706

i
 Biofloc technology and its potentiality for higher production of fish in Bangladesh1

By

Rabeya Chowdhury Shapla2

ABSTRACT
Biofloc technology (BFT), the new “blue revolution” in aquaculture, could be promising in
attaining it’s sustainability to aquaculture production without sacrificing the quality. It is mainly
based on the principle of waste nutrients recycling, in particular nitrogen, into microbial biomass
that can be used in situ by the cultured animals or be harvested and processed into feed
ingredients. A rearing of mono-sex GIFT tilapia attained a significantly (P<0.05) difference in
survival rate of fish among the treatments. Another study evaluates the biofloc technology (BFT)
in light-limited tank culture of Nile tilapia (Oreochromis niloticus). Two biofloc treatments and
one control were managed in indoor tanks: BFT fed a diet of 35% crude protein (CP), BFT fed a
diet of 24% CP, and clean water control without biofloc with 35% CP. BFT . Fish survival was
100%. Net fish production was 45% higher within the BFT tanks than within the control tanks
confirming the use of biofloc by fish as food. The study of evaluating the effect of biofloc
technology (BFT) application on water quality and production performance of tilapia
(Oreochromis sp.) at different stocking densities. The highest TAN and nitrite-nitrogen were
observed in control treatment at stocking density of 100 fish/m3 (3.97 mg TAN/L and 9.29 mg
NO2-N/L, respectively). The highest total yield was observed on top of things treatment at the
very best density treatment (43.50 kg). An experiment was conducted to research the expansion,
digestive activity of raising genetically improved farmed tilapia (GIFT) during a (RAS) and an
inside (BFT) system. At the top of the study, the individual fish weight at harvest was 22%
higher within the BFT fish than within the RAS fish.

Keywords: Biofloc, nutrients recycling, microbial biomass, TAN

1
A seminar paper for the course GFB598, Summer’ 20.
2
MS student, department of Genetics & Fish Breeding, BSMRAU, Gazipur 1706.

ii
 LIST OF CONTENTS
Sl. Topic Page no.
No.

01 Abstract ii
02 List of contents iii
03 List of tables iv
04 List of figures v
05 CHAPTER 1: Introduction 1-2
06 CHAPTER 2: Materials and methods 4
07 CHAPTER 3: Review of Findings 5-21
08 CHAPTER 4: Conclusion 22
09 References 23-29

iii
 LIST OF TABLES

Sl. No. Name of the Tables Page No.

01 Proximate composition of biofloc 06

02 Some of the study conducted in fish with reference 07

to (BFT) based culture Systems
03 Mean values (±SE) of water quality parameters 09
observed throughout the study period
04 Growth performance of Nile tilapia in activated 14
suspension and clean water tanks fed
different levels of protein feed

05 Growth performance and feed utilization of tilapia 15

in re-circulating aquaculture system and indoor
biofloc technology system.

06 Effect of BFT and BFT with 10% WE on growth 16

performance

07 Effect of five carbon: nitrogen ratios in the 18

performance of Nile tilapia Oreochromis niloticus
in a biofloc system for six months, in comparison
with a standard method of cultivation (control).

iv
 LIST OF FIGURES

Sl. No Name of the Figures Page No.

01 Overview of possible parameters of bioflocs 02

technology and their probable effects.
02 Components needed for biofloc preparation 05
03 Scheme of biofloc technology in pond 06
04 Nitrogen and total phosphate dynamics in the RAS 12
and BFT tanks. RAS: Recirculating Aquaculture
System; BFT: Biofloc Technology.
05 Effect of BFT and BFT with 10% WE on nitrate of 13
Nile tilapia, O. niloticus, fingerlings rearing tanks.
06 19
Effect of different biofloc starters on ammonia, nitrate,
and nitrite concentrations in the cultured tilapia O.
niloticus system.

07 Specific activities of protease and amylase in digestive 21

glands, stomachs and intestines of juvenile
Litopenaeus vannamei in the control and two bioflocs
treatments with two C/N ratios (15, 20) at the end of
30-day feeding experiment.

v
 CHAPTER 1

INTRODUCTION

With almost seven billion people on world, the requirements for aquatic food which is increasing
accordingly and hence, expansion and intensification of aquaculture production are highly
required. Aquaculture as a food‐producing sector offers many opportunities to remove poverty,
hunger and malnutrition, generates economic growth and ensures better use of natural resources
(FAO, 2017)

Biofloc technology (BFT) offers benefits in improving aquaculture production that could
contribute to the achievement of sustainable development goals. This technology may result in
higher productivity with less impact to the environment. Biofloc technology is mainly based on
the principle of waste nutrients recycling, in particular nitrogen, into microbial biomass that can
be used in situ by the cultured animals or can be harvested and processed into feed ingredients
(Avnimelech, 2009; Kuhn et al., 2010). Heterotrophic microbiota is stimulated to grow by
steering the C/N ratio in the water through the modification of the carbohydrate content in the
feed or by the addition of an outer carbon source in the water (Avnimelech et al., 1999), so that
the bacteria can assimilate the waste ammonium for new biomass production. Hence,
ammonium/ammonia can be maintained at a low and non‐toxic concentration so that water
replacement is no longer required. Biofloc technology enhances the production and productivity
by its contribution to the supply of good quality fish juveniles, the latter being one of the most
important inputs in the production. In addition, it contributes to the improvement of the fish
production. In relation to the former, biofloc technology could support the supply of good quality
seeds by improving the reproductive performance of aquaculture animals and by enhancing the
larval immunity and robustness (Ekasari et al., 2015; Ekasari et al., 2016 and Emerenciano et al.,
2013)

This systems can also be developed and conduct in integration with other food production, by
advancing productive integrated systems, focusing at producing more food and also be feed
from the same area of land with less input. The biofloc technology is still in its developing
stage. More research is needed to optimize the system (in relation to operational parameters) e.g.

1
in relation to nutrient recycling. In addition, research findings will need to be shared with
farmers as the implementation of biofloc technology will require upgrading their skills.

Biofloc systems were also helped to prevent the introduction of disease to a farm from incoming
water. In the past, standard operation of shrimp ponds included water exchange (typically 10
percent per day) used as a method to control water quality. In estuarine areas with many shrimp
farms go through water exchange, disease would spread among farms. Reducing water exchange
is an obvious strategy for developing farm biosecurity. Shrimp farming moving toward more
closed and intensive production where waste treatment is more inter-analyzed. Some types of
biofloc systems have been used in commercial aquaculture or evaluated in research. The two
basic types are those that are exposed to natural light and those that are not exposed. One of this
systems exposed to natural light include outdoor, lined ponds or tanks and lined raceways for
shrimp culture in greenhouses. A complex mixture of algal and bacterial composition control
water quality in such “greenwater” biofloc systems. Maximum biofloc systems in commercial
use are greenwater. However, some biofloc systems (raceways and tanks) have been installed in
closed buildings with no contact to natural light. These systems are worked as “brown- water”
biofloc systems, where only bacterial processes control water quality.

Fig 1: Overview of possible parameters of bioflocs technology and their probable effects. (crab,
2010)

2
 OBJECTIVES
The study has undertaken to accomplish the following objectives;
 The general objective of this study was to explore the possible contribution of biofloc
technology application to aquaculture production, while maintaining sustainable
practices,
 Aquaculture production can be optimized by, the maintenance of water quality by the
uptake of nitrogen compounds,
 Providing an optimum nutrition to ensure maximum growth as well as increasing culture
feasibility by reducing feed conversion ratio and a decrease of feed costs.

3
 CHAPTER 2

MATERIALS AND METHODS

This seminar paper is exclusively a review paper. All of the information has been collected from
the secondary sources. The study was carried out based on the information through review of
related thesis, journals, reports and books. The necessary data were collected from source like
internet, National Fish week compendiums, different annual statistical yearbooks of Bangladesh,
newspapers, watching with different on-going researches in YouTube. I got suggestion and
valuable information from my major professor and my course instructors. I myself compiled the
collected information and prepared this seminar paper.

4
 CHAPTER 3

REVIEW OF FINDINGS
3.1. Basic view of biofloc
3.1.1 Essential component of Biofloc preparation
In general, biofloc is the macro-aggregation of bacteria, algae, detritus and other decomposed
components (Avnimelech et al., 1994). It is the combination of bacteria, diatoms, zooplankton,
protozoa, macro-algae, feces, uneaten feed (Fig. 2), and exoskeleton from dead organisms
(Decamp et al., 2008). It is a group of biotic and abiotic particulate components suspended in the
water which includes bacteria, planktons, and other organic materials (Hargreaves et al., 2006)

Fig 2: Components needed for biofloc preparation (Source: Daniel N and P. Nageswari 2017)

3.1.2. Nutritional quality of biofloc

The biofloc samples were analyzed for Kjeldahl nitrogen (Kj-N), total ammonia nitrogen (TAN),
total suspended solids (TSS) and volatile suspended solids (VSS). These properties were
determined following APHA (1998). The difference between Kjeldahl-N and TAN was used to
calculate the protein content of the bioflocs by multiplying the organic nitrogen content by 6.25
(Jauncey, 1982). The ash content was determined using TSS and VSS values. Lipids were
extracted according to Folch et al. (1957), using the modification of Ways and Hanahan (1964).
Protein, lipid and ash content were expressed as percentage of the dry weight (% DW) of the
bioflocs.

5
The total carbohydrate was calculated according to the following formula: carbohydrate (%
DW) = 100 – (crude protein (% DW) + lipid (% DW) + ash (% DW)) (Manush et al., 2005). The
gross energy content of the diets was calculated using kilo joule (kJ/g DW) values of 23.0, 38.1
and 17.2 for protein, lipid and carbohydrate respectively in (Table. 1) (Tacon, 1990).

Table. 1: Proximate composition of biofloc.

Component Crude amount
Protein 23.0 (kJ/g DW)
Lipid 38.1 (kJ/g DW)
Carbohydrate 17.2 (kJ/g DW)

3.1.3. Biofloc preparation

If nitrogen and carbon are well balanced in the solution, ammonium in addition to organic
nitrogenous waste will be converted into bacterial biomass (Schneider et al., 2005). By adding
carbohydrates to the pond, heterotrophic bacterial growth is stimulated and nitrogen uptake
through the production of microbial proteins takes place (Avnimelech et al., 1999) (Fig: 3). The
microbial biomass yield per unit substrate of heterotrophic bacteria is about 0.5 g biomass C/g
substrate C used (Eding et al., 2006) .Suspended growth in ponds consists of phytoplankton,
bacteria, aggregates of living and dead particulate organic matter, and grazers of the bacteria
(Hargreaves et al., 2006).

Fig 3: Scheme of biofloc technology in pond (Source; Google)

6
3.1.4. Benefits of Biofloc Technology (BFT) system
 Eco-friendly culture system.
 It reduces environmental impact.
 Improves land and water use efficiency
 Limited or zero water exchange
 It enhances growth performance, survival rate, and feed conversion in the culture
systems.
 Higher bio-security.
 Reduces water pollution and the risk of introduction and spread of pathogens
 Cost-effective feed production.
 It lowered utilization of protein rich feed and cost of standard feed.
 It decreases the pressure on capture fisheries i.e., use of cheaper food fish and trash fish
for fish feed formulation.

Table 2: Some of the study conducted in fish with reference to (BFT) based culture Systems
SL. No. Species studied Results acquired in the study with (BFT)

01 Labeo rohita Reduced the artificial feed reliance and improved

the utilization of bioflocs as feed to 50% (Sharma
et al., 2015)
02 Oreochromis niloticus Fish survival was 100% and results in biofloc
utilization as food (Azim and Little, et al., 2008)
03 Oreochromis sps. Improvement in the water quality, fish survival and
minimization in the external feed requirement
(Sharma et al., 2015)
04 Litopenaeus vannamei Promoted the animal growth, health, digestion and
feed utilization performances (Xu et al., 2012)
05 Penaeus monodon Gave the beneficial effects on growth performances
and digestive enzyme activities (Anand et al., 2013)
06 Litopenaeus vannamei Increase in 30% growth and survival of shrimp in
Biofloc treatment (Piedrahita, 2003)

7
3.2. Applications and results

3.2.1. Physico-chemical parameter of water in (BFT)

It is considered that most of the uneaten feeds that present in the water damage the pond water
and threaten the animals to disease susceptibility (Francis-Floyd et al., 2009). It was
demonstrated in the earlier findings that adopting biofloc technology would solve the problems
concerned with ammonia toxicity, with the increasing consumption of nitrogen by heterotrophic
bacteria the nitrification process advances, which ensures the reduction in the concentration of
ammonium in the culture systems (Hargreaves, 2006). The study also demonstrated that the
production rate for heterotrophic bacteria for the utilization of ammonium is 10 times greater by
heterotrophic bacteria as compared to that of nitrifying bacteria (Hargreaves, 2006)

A study was conducted by (Nahar and Bakar et al., 2015), where the values of water quality
parameters such as water temperature, dissolved oxygen, pH and transparency in different
treatments are shown in (Table. 3). The mean values of water temperature in different treatments
were 23.33±1.04, 23.21±1.06, 23.10±1.03 and 23.15±1.03°C in CF, WB, BFT and RWB,
respectively (Table 3). The highest (29.91°C) and lowest (17.86°C) water temperature in the
present study might be due to the bright sunshine and cold weather. The amount of dissolved
oxygen in the water is directly influenced by temperature and also affects the cultured species
metabolism, which determines fish growth aspects. In BFT set ups, an intermediate water
temperature of 20-25°C could be best to obtain stable flocs proposed by Craig and Helfrich.
Tilapia (O. niloticus) didn't grow at temperature below 16°C and didn't survive at temperature
below 10°C for quite a couple of days. In the present study, the day temperature within the ponds
didn't fall below 17.86°C and fish probably didn't stop eating. Dissolved Oxygen (DO) varied
from 4.25 to 6.10 mg/l with mean values of 5.18±0.17, 5.05±0.16, and 4.83±0.17 and 4.92±0.19
mg/l in CF, WB, BFT and RWB, respectively

8
Table.3. Mean values (±SE) of water quality parameters observed throughout the study period
Water quality CF WB BFT RWB
parameters
Temperature 23.33±1.04 23.21±1.06 23.10±1.03 23.15±1.03
(°C)
Dissolved 5.18±0.17 5.05±0.16 4.83±0.17 4.92±0.19
oxygen (mg/l)
pH 7.00±0.03 7.06±0.04 7.10±0.05 7.08±0.05
Transparency 34.24±0.25 29.86±0.01 26.06±0.39 31.06±0.30
(cm)

According to Luo et al., (2013) in the BFT tanks, the TAN (Total Ammonia Nitrogen) was
primarily from the decomposition of organic matter such as fish waste and dead microbes in the
reactors. The TAN (Total Ammonia Nitrogen) can be rapidly taken up and stored by the biofloc
microbes by adding organic carbon to stimulate the growth of heterotrophic bacteria. However,
the packaging of nitrogen in microbial cells is temporary because cells turn over rapidly and
release nitrogen as NH3 when they decompose. Therefore, the TAN (Total Ammonia Nitrogen)
was repeatedly cycled between the dissolved NH3 and the flocs solids, exhibiting an obvious
fluctuation (Fig. 4A). The peaks of the TAN and NO2−-N occurred on day 20 during the BFT
system start-up. The highest concentrations of TAN and NO2−-N were 60 ± 0.45 mg L−1 (Fig.
4A) and 119 ± 2.01 mg L−1 (Fig. 4B), respectively. With the acclimation of the system, the TAN
and NO2—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 accumulation of NO3−-N was observed in the
culture tanks during the entire experiment (Fig. 4C). The safe level of NH3 for larval tilapia
(Oreochromis niloticus) is 0.42 mg L−1 (Benli and Köksal, 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 L −1,

9
which is higher than the highest TAN in the current study. Therefore, it can be concluded
that our BFT system is NH 3 and toxicity-free. NO2 −-N is an intermediate during both
nitrification and denitrification (Chuang et al., 2007; Ruiz et al., 2003). The accumulation of
NO2−-N is common in intensive aquaculture (Wang et al., 2004), probably due to the free
NH3 inhibition during nitrification and denitrification (Shi et al., 2011). Based on a
recommendation from Wuertz et al., (2013), salt was added to the BFT tanks (1%, w/w −1 ) to
compensate for the stress fromNO 2−-N to the tilapia in the current experiment. The NO 2 −-N
concentration in the BFT tanks was much higher than that from other reports (Asaduzzaman
et al., 2009; Xu and Pan, 2012), which is probably because of the higher stocking density in
the current experiment. There is no accumulation of NO 3 −-Nin the BFT tanks perhaps
because the process of nitrification might be inhibited by the high C: N ratio, the NO 3 −-N
was used by the heterotrophic bacteria (Luo et al., 2013), or denitrification 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 fish
and that a relatively large fraction (80%–90%) is released into the fish culture systems
(Barak et al., 2003). A targeted method for phosphorus removal in the current RAS was not
3−
developed; therefore, the accumulation of PO 4 -P in the RAS tanks was 45 ± 1.23 mg L −1
3−
at the end of the experiment (Fig. 4 D), whereas the PO 4 -P level in the BFT tanks was as
low as 4.01 ± 0.34 mg L−1, which was probably because the growth and reproduction of the
biofloc microorganisms assimilated the phosphorus that was not used by the fish.

10
Fig 4: Nitrogen and total phosphate dynamics in the RAS and BFT tanks. RAS: Recirculating
Aquaculture System; BFT: Biofloc Technology. (Luo and Gao et al., 2013)

In Abduljabbar et al., (2015) analyzed and compared the changing of TAN concentrations was
fairly consistent over time among the investigated groups with tendency to decrease with BFT
treatments than the control group (Fig. 5). The TAN were controlled in BFT treatments or BFT
with 10% WE via carbon source (molasses) addition to stimulate the growth of heterotrophic
bacteria, TAN can be used and stored by the formation of BFT microbes (Avnimelech et al.,
1999; Ebeling et al., 2006), which may explain why the TAN concentration did not exhibit
significant differences between the clean water control group and the BFT treatments.

The nitrate (NO3-N) reached each beak after four weeks of treatments for all treatments and
decreased gradually over time (Fig. 5). The least nitrate level was observed with BFT with zero
WE. The accumulation of nitrate during the first weeks may have been caused by nitrification
processes, which are common in BFT systems (Azim and Little, 2008 and Zhao, et al., 2012).
The subsequent reduction of nitrate in the BFT treatments likely occurred due to immobilization
by heterotrophic bacteria, which inhibited the nitrification and de-nitrification process (Azim and

11
Little et al., 2008). Also, Luo et al. (2014) reported that nitrate did not accumulated in the BFT
tanks perhaps because the process of nitrification might be inhibited by the high C:N ratio, and
used by the heterotrophic bacteria or de-nitrification occurred in the system.

Fig 5: Effect of BFT and BFT with 10% WE on nitrate of Nile tilapia, (O. niloticus), fingerlings
rearing tanks.
BFT: Biofloc technology; WE: Water exchange.

3.2.2. Growth performance and fish yield parameters

A study was carried out by Azim and Little et al., (2008) to evaluate growth and fish yield
parameters of Tilapia are given in (Table.4). Tilapia survival was 100% in all treatment and
control tanks. Individual fish weight at harvest was 9–10% higher in the BFT treatments than in
the control. The BFT treatments contributed 44–46% greater individual weight gain and net fish
production than those in the control confirming the utilization of biofloc by fish as food. Most of
the tilapias are known to utilize in situ produced food particles including suspended bacteria
(Beveridge et al., 1989). Avnimelech (2007) has also confirmed that, the biofloc uptake by
Mozambique tilapia using stable nitrogen isotope labeling technique. However, there was no
significant difference in fish growth/production between 35% and 24% CP fed tanks under the
BFT. The FCR value was significantly higher in the control than in the BFT treatments.
Little et al., (2008) reviewed that the final biomass levels of 10–28 kg fish m−3 achieved in
indoor and outdoor BFT systems throughout the year.

12
A study was carried out by Azim and Little et al., (2008) to evaluate growth and fish yield
parameters of Tilapia are given in (Table.4). Tilapia survival was 100% altogether treatment and
control tanks. Individual fish weight at harvest was 9–10% higher within the BFT treatments
than within the control. The BFT treatments contributed 44–46% greater individual weight gain
and net fish production than those in the control confirming the utilization of biofloc by fish as
food. Most of the tilapias are known to utilize in situ produced food particles including
suspended bacteria (Beveridge et al., 1989; Beveridge and Baird, 2000).
Avnimelech et al., (2007) has also confirmed that, the biofloc uptake by Mozambique tilapia
using stable nitrogen isotope labeling technique. However, there was no significant difference in
fish growth/production between 35% and 24% CP fed tanks under the BFT. The FCR value was
significantly higher in the control than in the BFT treatments. Although there was clear evidence
that the biofloc significantly contributed to the expansion and production of fish. Little et al.,
(2008) reviewed that the final biomass levels of 10–28 kg fish m−3 achieved in indoor and
outdoor BFT systems throughout the year.

Table. 4 Growth performance of Nile tilapia in activated suspension and clean water tanks fed
with different levels of protein feed

Parameters 35% CP without 35% CP with 24% CP with

biofloc biofloc biofloc
Initial individual 99.61±13.74 100.69±13.61 98.45±12.71
weight (g)
Final individual weight 127.51±28.17 140.72±27.26 138.58±24.99
(g)
Survival (%) 100 100 100
Individual weight gain 27.9±0.69 40.04±3.04 40.08±4.34
(g)
Net yield (kg m−3) 3.35±0.08b 4.80±0.60 4.90±0.59
FCR 4.97±0.12 3.51±0.44 3.44±0.45

13
In the study of Luo and Gao ( 2013), Growth performance and feed utilization of fish and fish
yield parameters are described in (Table.5). Even when the water quality was not stable and there
were high fluctuations in the concentrations of TAN and NO2−-N, no fish died during the entire
experiment, and the fish in the BFT tanks grew more satisfactorily than those in the RAS tanks
did. The individual fish weight at harvest was 22% higher within the BFT fish than within the
RAS fish. The total weight gain and SGR of the BFT fish were, respectively, 129% and 12%
above those of the RAS fish. The FCR value in BFT treatment fish was 1.20 ± 0.03, 18% better
than that of the RAS fish, which had an FCR of 1.47 ± 0.02.

Table.5. Growth performance and feed utilization of tilapia in recirculating aquaculture system
and indoor biofloc technology system.

Parameters RAS BFT

Initial number (fish tank−1) 100 100
Initial mean weight (g 24.17 ± 2.49 24.17 ± 2.49
fish−1)
Final mean weight (g fish−1) 138.29 ± 34.61 168.58 ± 40.64
Survival rate (%) 100% 100%
Specific growth rate (% 1.90 ± 0.30 2.13 ± 0.29
day−1)
Weight gain (%) 472.14 ± 143.18 597.48 ± 168.16
Total weight gain (kg m−3) 28.87 36.95

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A treatment was run by Abduljabbar et al., (2015) for the comparison in growth performance of
fishes between BFT treatments and BFT with 10% WE. The growth performance of fish in BFT
treatments and BFT with 10% WE was found higher significantly (P <0.05) than fish in the
control group in the case of (final body weight, weight gain, average daily gain and specific
growth rate). The increase of final weight was 28.70 and 22.20% with BFT or BFT with 10%
WE, respectively, than in the control group. Survival rate was found high (98-99%) with all
conducted treatments, where rearing fish under BFT technology with zero WE or limited (10%)
WE did not negatively affect the survival rate (Table.6). Moreover, the present results verified
that intensive tilapia production in the BFT systems increase feed and nutrients utilization
significantly (P <0.05) compared with in the control.

According to Krummenauer et al., (2008) shrimp reared in the environment with molasses
addition exhibited a survival rate, final weight and specific growth rate (SGR) significantly
higher than those of the control did.

Table.6. Effect of BFT and BFT with 10% WE on growth performance

Item Control BFT BFT + 10% WE

Initial weight (g/fish) 3.38±0.02 3.44±0.02 3.37±0.03
Feed intake (g/fish 20.16b±0.15 22.67a±0.23 22.19a±0.24
Weight gain (g/fish) 8.78b±0.26 12.22a±0.38 11.49a±0.27
Final weight (g/fish) 12.16b±0.25 15.65a±0.36 14.86a±0.28
ADG (g/fish/day) 0.07b±0.0 0.10a±0.0 0.10a±0.0
SGR (%/day) 1.07b±0.02 1.26a±0.02 1.24a±0.01
Survival rate (%) 98±0.82 99±0.67 98±1.11

*Means followed by different letters in the same raw differ significantly (P.0.05).

BFT: Biofloc technology; WE: Water exchange; ADG: Average daily gain; SGR: Specific
growth rate; FCR: Feed conversion ratio; PER: Protein efficiency ratio; PPV: Protein productive
value.

15
3.2.3. Maintaining C/N ratio

The maintenance of C/N ratio is quite prerequisite for controlling of accumulating organic
nitrogen and for the production of microbial communities in the water (Asaduzzaman et al., 2008
and Emerenciano et al., 2012). The inorganic nitrogen is converted into organic nitrogen when
C: N ratio is sufficient to produce bacterial cells (Aly et al., 2008). As carbohydrate is involved
in the part of respiration process, during aerobic situations the condition of C: N ratio must be
more than bacterial body compositions (Emerenciano et al., 2013). It was found that around 10
mg NH4+-N/L can be completely absorbed when glucose was added as a substrate and when the
maintenance of C/N ratio was 10:1. To minimize the synthetic feed requirement, the practice of
accelerating C: N of upper than 10:1 by utilizing different low-cost carbon sources which are
locally obtainable is common in biofloc waters (Crab et al., 2010). Apart from reducing the feed
cost, utilization of biofloc components will also decrease the amount of protein in the feed
(Avnimelech, 1999; Hargreaves, 2006). It was established that the buildup of toxic inorganic
components including, NH4+ and NO2- are going to be stopped within the water when the
upkeep of C/N ratio is high in the biofloc system because the ammonium consumption by the
microbial community.

By Jorge et al., (2015), survival of juvenile tilapia in the biofloc treatments was similar and
significantly higher (94.60 ± 2.03%) than survival in the control (84.96 ± 1.53%) (Table.7)
During the first three months, exponential growth occurred in all tilapia. From months 4–6,
tilapia raised in the control had a significantly higher growth rate than tilapia in the biofloc
treatments. The high final weight of tilapia in the control treatment was a response to lower
density caused by higher early mortality (r = 0.81 and P b 0.001). The highest final weight,
weight gain, weight gain per day, and final biomass was in the 10:1 and 15:1 C: N treatments,
with no significant differences with tilapia in treatment 12.5:1. The lowest final weight and
weight gain occurred in the 17.5:1 and 20:1 C: N treatments. The feed conversion ratio in all
tilapia in the biofloc treatments was close to 1.06 ± 0.05, with consumption of feed of 1.60%
each day in all treatments and the control. Ranking of filet yield of the treatments was: 10:1 N
15:1 N control N 12.5:1 N 17.5:1 N 20: 1 (Table 7).

16
Table.7. Effect of five carbon: nitrogen ratios in the performance of Nile tilapia O. niloticus in a
biofloc system for six months, in comparison with a standard method of cultivation (control).

Variable Control 10:1 15:1 17.5:1 20:1

S (%) 84.96 ± 1.53 94.75 ± 1.53 94.28 ± 2.08 94.18 ± 1.53 94.32 ±
1.53
IW (g) 38.4 ± 1.49 38.4 ± 1.49 38.4 ± 1.49 38.4 ± 1.49 38.4 ± 1.49
FW (g) 280.89 ± 254.31 ± 20.50 251.56 ± 15.76 233.31 ± 230.66 ±
5.43 10.87 10.38
IGW (g) 223.69 ± 196.26 ± 29.54 193.52 ± 23.91 176.12 ± 172.62 ±
21.03 22.44 19.88
WGD 1.24 ± 0.11 1.09 ± 0.16 1.07 ± 0.13 0.97 ± 0.12 0.95 ± 0.11
(g/d)
FCR 1.02 ± 0.01 1.01 ± 0.01 1.03 ± 0.01 1.13 ± 0.01 1.14 ± 0.01
FB 17.86 ± 0.16 18.03 ± 0.25 17.73 ± 0.18 16.44 ± 0.04 16.28 ±
(kg/m3 ) 0.02
FP (%) 29.53 ± 1.50 32.24 ± 1.83 31.62 ± 1.84 27.21 ± 1.10 26.77 ±
1.95

Different letters in the same row represent significant differences at P b 0.05. FW = final weight,
FL = final length, IGW = individually gain weight, WGD = weight gained per day, FCR = feed
conversion rate, FB = final biomass, FP = filet production.

17
The study by Putra I. et al., (2010), revealed that the concentrations of ammonia, nitrite, and
nitrate were significantly lower within the biofloc system using starters compared with those
within the system without a starter (control). Biofloc has probiotic bacteria which will change
ammonia to nontoxic materials (such as nitrate) that are useful for phytoplankton growth.
Therefore, the ammonia and nitrate concentrations are low in the culture media (Hargreaves et
al., 2005). Biofloc doesn't necessarily only contain bacteria (for example, Bacillus), but is
additionally composed of other useful microorganisms like microalgae and zooplankton that are
trapped by organic particles (Crab, 2007). Algae and zooplankton can be used by cultured biota
(tilapia) as natural food.

Fig 6: Effect of different biofloc starters on ammonia, nitrate, and nitrite concentrations in the
cultured tilapia system

(a) Concentrations of ammonia, (b) nitrate, and (c) nitrate during 40 days of the experiment. A =
control treatment, B = molasses starter, C = tapioca starter, and D = sucrose starter.

18
In general, the starters used in this study were carbohydrate compounds. However, the study
showed that the molasses starter yielded slightly better results compared with the other starters.
This finding indicated that molasses was the best carbon source for biofloc in the tilapia culture
system. Molasses can provide a sufficient carbon level for heterotrophic bacteria that use this
carbon as an energy source for growth (Avnimelech et al., 2009). Molasses are a liquid
byproduct from the sugar industry. This material features a total carbon content of around 37%.
Therefore, molasses are rapidly soluble in water and can be quickly absorbed by heterotrophic
bacteria. In terms of chemical structure, molasses are classified as an easy carbohydrate
containing six C atoms (monosaccharides), while sucrose (treatment D) may be a combination of
two monosaccharides that contains 12 C atoms (sucrose). Tapioca is classified as a complex
carbohydrate (60,000 C atoms) and is more slowly digested by bacteria than molasses (Azhar,
2013).

3.2.4. Digestive enzyme activity during biofloc culture system

In the current study of Xu and Qing Pan et al., (2012), protease and amylase activities of the
shrimp in three groups were tissues-specific. Both enzymes activities were found highest in the
digestive gland, intermediate in the stomach, and lowest in the intestine. Although not
statistically significant, enhanced protease and amylase activities in the digestive glands of the
shrimp were observed in both biofloc treatments (Fig. 6A–B). Protease and amylase activities in
the stomachs of the shrimp in both biofloc treatments were significantly higher than those in the
control (Fig. 6C–D); while a decreasing trend of protease and amylase activities in the intestines
of the shrimp in both biofloc treatments was observed (Fig. 6E–F). A general enhancement in
protease and amylase activities of the shrimp in both biofloc treatments was observed, though the
effect of the bioflocs on each enzyme activity performed inconsistently among different digestive
tissues: digestive gland, stomach and intestine.

19
Fig 7: Specific activities of protease and amylase in digestive glands, stomachs and intestines of
juvenile L. vannamei in the control and two bioflocs treatments with two C/N ratios (15, 20) at
the end of 30-day feeding experiment. Means±S.E. indicated. n=4. Means within the same tissue
with different letters are significantly different (Pb0.05).

20
3.3. Potentiality of Biofloc technology (BFT)

This technology is essentially of zero water exchange oriented i.e. water exchange isn't required
within the culture ponds; therefore it required less water input which isn't only economical to the
farmers, but these also will minimize the pathogenic entry of animals through water and certify
for more biosecurity within the fish culture.
It also promises the less environmental impacts and footprints (Wasielesky et al., 2006).This
technology allows the animals to rear under the upper stocking density with effective feed
management (Crab et al., 2010; Crab et al., 2012).
The requirement for the feed is considerably less, as biofloc itself will be a feed for the cultivable
animals, which results in the lower FCR (Aiyushirota , 2009; Krummenauer, 2011; Perez-
Fuentes et al., 2013). Therefore, application of the technology will reduce the feed cost to the
farmers.
Biofloc increases the survival of fish since the beneficial microorganisms dominate in the biofloc
acts as an antagonism to the pathogenic bacteria which prevent the disease outbreak and expand
the percentage of survival during the harvest. This way (beneficial) bacteria present in the
biofloc prevent the colonization of any harmful bacteria that ensure the highest survival rate of
the fish in the farms (Megahed, 2010; Samocha, 2007; Perez-Fuentes et al., 2013).
Biofloc bacteria produce the poly hydroxyl butyrate (PHB) which is beneficial in the digestion
and metabolism of fatty acid and growth increment to the fish (De. Schryver et al., 2012).
Biofloc waters rich within the heterotrophic bacteria which utilize the toxic nitrogenous matters
as a substrate for his or her growth that helps maintaining the water quality through reducing the
organic loads as well as biochemical oxygen demand of the system (Avnimelech, 1994; Burford
et al., 2004).
Bioflocs comprise a wide assemblage of bacteria, algae, protozoa and other zooplankton
organisms, perhaps as many as 1000-2000 different species. As yet we do not know enough
about the composition of the bioflocs, nor our ability to affect it and the different effects it may
have on fish production and on the eco-stability of the system. It has been shown that the
immune systems of shrimp are enhanced in the presence of bioflocs and there is a lower
incidence of diseases among shrimp grown in biofloc systems.

21
It has demonstrated the probiotic effects of bioflocs against Streptococcus infection in tilapia.
Research on the effects of bioflocs on diseases is actively ongoing, and we can expect getting
more on how to use this system to control diseases.
Interesting new results demonstrate the effects of bioflocs on the fecundity of both shrimp and
tilapia: in both cases the number of eggs per female was about doubled. We do not know exactly
the mechanism of this effect. It is possibly caused by the top quality of the biofloc feed
components, better water quality, or the presence of hormones (or of components having
hormonal effects).
Biofloc technology has become a common way of running hatcheries and nurseries. Moreover,
biofloc systems are environmentally friendly due to the fact that there is almost no release of
nutrient rich drainage water to the environment.

22
 CHAPTER 4

CONCLUSION
Biofloc technology offers benefits in improving aquaculture production that would contribute to
the achievement of sustainable development goals. This technology could result in higher
production capacity with less impact to the environment. Furthermore, biofloc systems are often
developed and performed in integrated production with other food, thus enhancing productive
integrated systems, with the aim of manufacturing more food and feed from the same area of
land with fewer input. The biofloc technology remains in its infant stage. A lot more research is
needed to optimize the system; in relation to operational parameters e.g. in relation to nutrient
recycling, production and immunological effects. In addition, research findings will get to be
communicated to farmers because the implementation of biofloc technology would require
upgrading their skills. This review paper has showed the growth, digestive activity, welfare, and
water quality parameters of raising GIFT in RAS tanks and BFT tanks. At the beginning of the
study, the Initial mean weight (g fish−1) 24.17 ± 2.49 in RAS and 24.10 ± 2.44 for BFT treatment,
but at the end of the experiment, Final mean weight (g fish−1) was found 138.29 ± 34.61 in RAS
tanks and 168.58 ± 40.64 in the BFT tanks. The fishes flourished in both environments; however,
the BFT fish were superior to the RAS fish, and no mortality was found among the BFT fish,
although the accumulation of NO2−-N and TAN was observed in the tanks. The crude protein
content within the biofloc met the nutritional requirements of tilapia, but the crude lipid
concentration was quite low. A significant difference was found for lipase in the intestines and
stomachs of the fish cultured using both models; however, no substantial difference was found
for protease.
Various kinds of beneficial features can be ascribed to biofloc technology, from water quality
control to in situ feed production and some possible extra features. Biofloc technology offers
aquaculture a sustainable tool to simultaneous in its environmental, social and economical issues
concurrent with its growth. Researchers are challenged to further develop this technique and
farmers to implement it in their future aquaculture systems. The basics of the technology is there,
but its further development, fine-tuning and implementation will need further research and
development from the present and future generation of researchers, farmers and consumers to
form this system a keystone of future sustainable aquaculture.

23
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