Reviews in Aquaculture (2021) 13, 676–705 doi: 10.1111/raq.

12494

Use of biofloc technology in shrimp aquaculture: a

comprehensive review, with emphasis on the last decade
Abdel-Fattah M. El-Sayed
Oceanography Department, Faculty of Science, Alexandria University, Alexandria, Egypt

Correspondence
Abdel-Fattah M. El-Sayed, Oceanography
Department, Faculty of Science, Alexandria
University, Alexandria, Egypt. Email:
Abdelfatah.youssif@alexu.edu.eg
Received 26 April 2020; In Revised form 17 July
2020; accepted 31 July 2020.
Abstract
In face of the shortage of, and competition with, land and water, the sustainability
of aquaculture will have to depend on vertical development, through improving
production environments, increasing productivity and enhancing aquaculture
technologies. Biofloc technology (BFT) has emerged as new alternative for sus-
tainable aquaculture, which could contribute to FAO Sustainable Development
Goals (SDGs) related to food security. Extensive research has been carried out on
the development and application of BFT in aquaculture since early 1990s, with
emphasis on shrimp culture. Over 40% of BFT publications in aquaculture were
directed to shrimp farming. Therefore, I strongly believe that the accumulated
knowledge on the applications of BFT in shrimp farming and the experience
gained, especially during the last 10 years (2010–2020), are now more than wor-
thy of critical review and analysis. This review summarizes the most update
knowledge on the use of BFT in different marine shrimp and freshwater prawn
aquaculture. Emphasis has been on factors affecting shrimp production in BFT
systems, integration of biofloc-based shrimp farming with other aquatic farmed
species, nutritional value of bioflocs as a natural food or feed ingredient for
farmed shrimp and prawn, the application of BFT in different rearing phases, the
use of biofloc as a natural probiotics and their effects on shrimp health and physi-
ological functions, economic considerations and commercial applications of BFT-
based shrimp aquaculture, and the major challenges facing shrimp farming in bio-
floc systems.
Key words: application, Biofloc technology, C/N ratio, challenges, nutritional value, shrimp and
prawn, water quality.

resources, the sustainability of aquaculture will likely rely

Introduction
Feeding the growing human population, which is projected
to reach 9.6 billion by 2050, in face of the scarcity of natural
resources necessary for food production, such as land and
water, is a serious challenge. Aquaculture has emerged as
an ideal food production option in many countries. Sus-
tainable aquaculture development could contribute to dif-
ferent FAO Sustainable Development Goals (SDGs),
including SDG 1 (end poverty), SDG 2 (end hunger,
achieve food security and improve nutrition and promote
sustainable agriculture), SDG 8 (promote inclusive and sus-
tainable economic growth, employment and decent work
for all) and SDG 14 (conserve and sustainably use the
oceans, seas and marine resources for sustainable develop-
ment) (FAO 2017). Due to the shortage of land and water
on improving production environments, increasing pro-
ductivity, enhancing aquaculture technologies and reducing
production costs.
Biofloc technology (BFT) has been suggested as a sus-
tainable aquaculture system. The concept of BFT has been
recognized since early 1970s. However, extensive research
on the development and application of BFT has been car-
ried out since early 1990s, with encouraging results (Avn-
imelech 2015). According to National Agricultural Library
Glossary (United States Department of Agriculture), bio-
floc technology is defined as ‘the use of aggregates of bacte-
ria, algae or protozoa, held together in a matrix along with
particulate organic matter for the purpose of improving
water quality, waste treatment and disease prevention in
intensive aquaculture systems’. In other words, biofloc is a

676 © 2020 John Wiley & Sons Australia, Ltd


symbiotic process that includes confined aquatic animals,
heterotrophic bacteria and other microbial species in the
water. Through this process, ammonia is removed from
culture system, and the waste materials may be cycled into
supplemental food sources for farmed aquatic species
(Browdy et al. 2012). This means that BFT can be an ideal
option for sustainable and environmentally friendly aqua-
culture (Crab et al. 2012).
The contribution of shrimp culture to global aquaculture
output during the last decade has been significant. The pro-
duction of farmed shrimp and prawns has increased from
3,400,458 mt in 2008 to 6,004,353 mt in 2018 (76.6%
increase), representing 7.3% of global aquaculture produc-
tion, excluding aquatic plants, in 2018 (FAO 2020). White-
leg shrimp Litopenaeus vannamei is the most important
farmed single crustacean species. The production of this
species was almost doubled during the last 10 years (2008–
2018), contributing 53% to total farmed crustacean pro-
duction in 2018 (FAO 2020). This increase has been possi-
ble because of the impressive innovations and technical
developments in shrimp aquaculture in recent years. Out-
standing developments have been achieved in all shrimp
farming disciplines, including farming technologies, genet-
ics, hatchery management, nutrition, disease prevention
and combating, bio-security, harvesting and post-harvest
processing and marketing.
In recent years, special attention has been paid to biofloc
technology (BFT) as a promising tool for sustainable shrimp
aquaculture. Over 550 documents have been published on
the application of BFT in aquaculture, most of them
appeared during the past 10 years (2010–2020). Over 40%
of these publications were dedicated to shrimp aquaculture,
with white shrimp (L. vannamei) receiving most of the atten-
tion. More than 200 papers were published on raising this
species in biofloc systems. Research and development on
rearing shrimps in biofloc systems are also still escalating at
an outstanding rate. In 2019 only, 44 papers were published
on this subject in top, specialized journals. Fourteen papers
were also published in the first quarter of 2020.
Therefore, a comprehensive review and critical analysis
of the role of BFT in shrimp aquaculture appears timely.
This review will tackle this issue, through analysing and dis-
cussing the available information that has been published
on the application of BFT in shrimp aquaculture, particu-
larly during the last decade (2010–2020), with emphasis on
white shrimp (L. vannamei). It is hoped that this review
will benefit shrimp farmers, farm managers, researchers,
administrators and other relevant stakeholders. I also hope
they would appreciate the effort paid in bringing all this
diverse and scattered information to them in a single, com-
prehensive review.
As mentioned above, the BFT in shrimp and prawn
aquaculture has received considerable attention during the
Reviews in Aquaculture (2021) 13, 676–705
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Use of bioflocs in shrimp aquaculture
past decade (Panigrahi et al. 2017; Kumar et al. 2017; Kaya
et al. 2019). Published works covered various disciplines of
shrimp culture in biofloc systems, including the following
(i) types of biofloc systems (Panigrahi et al. 2018, 2019a,
2019b, 2020a); (ii) pathways for ammonia removal in bio-
floc systems (Kumar et al. 2019; Panigrahi et al. 2019c,
2020a); (iii) factors affecting shrimp production in BFT
systems; (iv) nutritional value of bioflocs for shrimp cul-
ture (Ekasari et al. 2014b; Khanjani et al. 2015; Panigrahi
et al. 2019d); (v) role of bioflocs as probiotics in shrimp
culture (Ekasari et al. 2014a; Anand et al. 2017; Panigrahi
et al. 2020c); (vi) application of BFT in shrimp culture
(Khanjani et al. 2017, 2020; Martinez-Porchas et al. 2020);
(vii) integrated biofloc-based shrimp farming (Fourooghi-
fard et al. 2018); (viii) economic considerations (Pinto
et al. 2020); and (ix) challenges facing BFT-based aquacul-
ture. The following sections will discuss and analyse these
studies.
Types of biofloc systems
The term ‘biofloc’ describes a number of aquaculture pro-
duction systems, where water quality is regulated by a com-
bination of autotrophic and heterotrophic microbial
processes. The role of phytoplankton metabolism (photo-
synthesis) and bacterial processes are essential in this pro-
duction system. A number of biofloc systems have been
developed, depending on farm site (indoor vs. outdoor),
farming intensity (semi-intensive, intensive or super-inten-
sive) and technical protocols adopted. In general, biofloc
systems can be divided into the following types.
Suspended-growth systems (without media)
The suspended-growth systems (SGS) is also known as: ‘al-
gae, bacteria, zooplankton and detritus (ALBAZOD)’ ‘pho-
tosynthetic suspended-growth aquatic system’, ‘organic
detrital algae soup (ODAS)’, ‘zero exchange, aerobic and
heterotrophic (ZEAH) culture system’, ‘aerated microbial
reuse systems’, ‘activated sludge ponds’, ‘suspended, bacte-
rial-based treatment process’ and ‘photosynthetic sus-
pended-growth system (PSG)’ (Hargreaves 2006). PSG
system is generally used for the production of large
amounts of microbial biomass. In this system, substrates,
such as organic carbon source, ammonia (NH3) and nitrite
(NO2), are required, together with heavy aeration, to keep
the substrates and microbial communities in suspension
and, in turn, increase the surface area for bacterial action.
This means that water quality is maintained through active
mass of phytoplankton, attached bacteria and other living
organism and particulate organic matter (Hargreaves
2006). During this process, phytoplankton, microbial flocs
and other associated organisms are consumed by farmed
677
A. M. El-Sayed
aquatic animals, leading to improvement of the system effi-
ciency and reducing production costs.
Greenwater culture system, which is generally adopted in
outdoor environments, is an efficient application of PSG
technology. In this system, NH3 and NO2 are oxidized into
nitrate (NO3) through nitrifying bacteria grown on sus-
pended organic matter. The bacteria remove the organic
matter from the tanks and use it as food. Vigorous aeration
is necessary to support the suspension of the microbial
community, maximize contact between bacteria and waste
products, and increase phytoplankton productivity. Phyto-
plankton dies, flocculates, and therefore, solid wastes
should be removed continuously.
Attached-growth biofiltration technology (with moving
media)
Attached-growth biofiltration technology (AGB) is also
known as ‘attached-growth membrane bioreactor
(AGMBR)’ (Hargreaves 2006; Ibrahim et al. 2019). In this
system, substrates are transported from rearing units to
specialized reactors to perform a specific operation. In
comparison with SGS, AGB is characterized by biofiltering
media with a high specific surface area. Therefore, nitrifica-
tion efficiency in AGB is much higher than in SGS. This
system can also reduce the suspended solids without limit-
ing the efficiency of the process (Hargreaves 2006).
Moving bed biofilm reactor
Moving bed biofilm reactor (MBBR) is an aerobic wastewa-
ter treatment process, based on plastic biocarriers on which
microbial biomasses attach and develop. The performance
efficiency of MBBR depends on the media supplied to the
reactor, surface area of the biocarrier, dissolved oxygen
(DO) and the organic loading (Ødegaard et al. 2000). MBBR
is an advanced hybrid of SGR and AGB, incorporating the
advantages of both systems and achieving higher concentra-
tion of biomass in the bioreactors. MBBR is highly efficient
in removing chemical oxygen demand (COD) (up to 90%)
and biochemical oxygen demand (BOD) (up to 95%).
Periphyton technology
Periphyton technology can also be used to remove organic
and inorganic wastes, enhance natural food production for
farmed organisms and improve the quality of culture water
(Azim et al. 2001; Tammam et al. 2020). In this technology,
sessile autotrophic and heterotrophic aquatic biota, com-
prising bacteria, fungi, protozoa, phytoplankton, zooplank-
ton, benthic organisms and a wide range of invertebrates,
are developed on submerged substrates and used as natural
foods for fish and shrimps (Azim et al. 2001). The efficiency
678
of periphyton biomass production depends on light inten-
sity and nutrient availability. Maintaining optimum C/N
ratio is also necessary for the optimization and enhance-
ment of periphyton production in pond-based aquaculture.
Periphyton is generally used for raising herbivorous,
omnivorous and filter-feeding fish and crustaceans, such as
tilapia (Tammam et al. 2020), mullets (Richard et al. 2010)
and freshwater prawn (Uddin et al. 2006).
These types of BFT systems can be applied in aquaculture
practices, with varying efficiencies in ammonia and solids
removal, biofloc production and aquaculture outputs. For
example, sequencing batch reactors (SBRs) and membrane
biological reactors (MBRs) can remove nutrients and solids
from aquaculture farm effluents (Kuhn et al. 2009, 2010).
SBRs generally require addition of carbon source, while
MBRs do not require carbon supplementation. In addition,
suspended-growth biological reactors (SGBRs) have been
used to produce bioflocs from food processing wastes
(Kuhn et al. 2016). Different reactors have also been
applied for treatment of wastewater and ammonia removal
from aquaculture effluents. They include rotating biological
contactors (RBCs), packed tower and fluidized bed reactor
(FBRs) (Miller & Libey 1985; Brazil 2006). RBCs are more
efficient in total ammonia nitrogen (TAN) removal than
FBRs and packed tower (Miller & Libey 1985).
In terms of natural light exposure, two BFT systems are
generally applied in aquaculture; outdoor (light exposed)
and indoor (no natural light exposure) (Hargreaves 2013).
Outdoor system, which is also known as ‘greenwater’ biofloc
system, is common in commercial aquaculture. It comprises
a mixture of algal and bacterial communities which can be
used as natural food for farmed aquatic animals (Kuhn &
Lawrence 2015). On the other hand, indoor BFT systems are
generally installed in closed areas, with limited light or even
no exposure to natural light. They are known as ‘brown-wa-
ter biofloc systems’, because only bacterial biomass is pro-
duced. Therefore, the uptake of bioflocs by farmed fish and
shrimp will depend mostly on the biofloc system adopted,
cultured species and size, floc size and density and culture
conditions (Avnimelech 2007; Crab et al. 2010).
Pathways for ammonia removal in biofloc systems
In BFT systems, ammonia is removed, and the waste mate-
rials may be cycled into natural food for farmed fish and
shrimp (Browdy et al. 2012). Typically, nitrogen conversion
includes three pathways for the removal of total ammonia
nitrogen (TAN) in aquaculture systems, they are (Ebeling
et al. 2006; Crab et al. 2010) as follows:
(1) Photoautotrophic removal by aquatic plants. In this pro-
cess, nutrients released into water effluents are
removed by mainly aquatic plants.
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(2) Autotrophic bacterial conversion of ammonia-nitrogen to
nitrate-nitrogen. In this pathway, autotrophic bacteria
and other autotrophs synthesize their cell constituents,
through photosynthesis, using carbon dioxide as a car-
bon source.
(3) Heterotrophic bacterial conversion of ammonia-nitrogen
directly to microbial biomass. In this pathway, the addi-
tion of organic carbon source is required for stimulat-
ing heterotrophic bacterial growth. At appropriate
carbon-to-nitrogen (C/N) ratio, these bacteria will
assimilate ammonia-nitrogen into cellular protein.
The three nitrogen conversion pathways are integral
components in the formation of bioflocs inside the culture
systems. These bioflocs can be used as natural foods source
for cultured organisms, in addition to improving the water
quality in the culture system.
Therefore, biofloc formation process includes the biolog-
ical nitrification (oxidation) of ammonia into nitrite (NO2)
by bacteria of the genera Nitrosomonas and Nitrosococcus,
which is further oxidized into nitrate (NO3) by Nitrobacter
and Nitrospira bacteria. Finally, nitrate can be reduced into
nitrogen gas through anoxic denitrification, mainly by
Achromobacter and Pseudomonas (Schramm et al. 1999).
However, the physical properties and composition of bio-
flocs vary considerably, depending upon the biofloc system
adopted, the quantity and quality of feed used, target spe-
cies, environmental conditions, type of aeration and factors
associated with floc production and colonization (De
Schryver et al. 2008; Ray et al. 2010a,b).
Factors affecting shrimp culture in biofloc systems
A number of biotic factors, including stocking size and den-
sity, supplemental feed and natural food availability have
been reported to affect biofloc production, efficiency and use
for shrimp aquaculture (Xu & Pan 2014a; Zhu et al. 2016;
Esparza-Leal et al. 2020). Abiotic factors, including tempera-
ture, oxygen, pH, alkalinity, light exposure, total nitrogen,
ammonia, nitrite and nitrate, also play a similar role in
shrimp production in BFT systems (Hostins et al. 2015;
Pi
erri et al. 2015; Khanjani et al. 2020). Some other culture
conditions, such as the addition of carbon source (to main-
tain appropriate C/N ratio), water exchange and rearing sub-
strates, are also of prime importance for optimal
performance of shrimp and prawn in biofloc systems
(Schveitzer et al. 2013a; Huang et al. 2017; Olier et al. 2020).
These factors are highlighted in the following sections.
Stocking density
Stocking density is a key factor affecting pond dynamics,
productivity and sustainability. Increasing stocking density
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Use of bioflocs in shrimp aquaculture
will require large amounts of feeds and result in excessive
nutrient loads, which need to be recycled by microbial
communities. These nutrients may cause eutrophication of
the systems, leading to development of microbial biomass
(Avnimelech 2007). Under these circumstances, feed wastes
and animal excretions are decomposed into ammonia,
which is assimilated by algae and heterotrophic bacteria to
build cellular proteins. Therefore, at high stocking density,
sufficient aeration is required to maintain a high particle
density. Also, the addition of organic carbon, to maintain
appropriate C/N ratio, is necessary to optimize bacterial
growth and production of microbial flocs.
Intensive shrimp culture is currently widely practised,
especially in South East Asia, with varying degrees of suc-
cess (Arnold et al. 2005; Thong 2014). The BFT is being
practised in semi-intensive commercial shrimp grow-out
ponds and also in super-intensive tank and raceway sys-
tems, with a production of over 9 kg shrimp m 3 (Taw
2010). Shrimp survival, growth performance and health
status generally decrease with increasing stocking density,
as reported for white shrimp Litopenaeus van-
namei (Wasielesky et al. 2013; Liu et al. 2017) and freshwa-
ter prawn Macrobrachium rosenbergii (Negrini et al. 2017).
This has been attributed mainly to increased competition
for the space and food sources, cannibalism and instability
in water quality (Abdussamad & Thampy 1994; Arnold
et al. 2005).
Carbon sources and carbon-to-nitrogen ratio
A classic biofloc system requires external inputs including
exogenous carbon source to maintain the appropriate car-
bon-to-nitrogen (C/N) ratio needed for microbial produc-
tion. A heterotrophic-based biofloc system uses the carbon
available as energy source and ammonia-nitrogen for cellu-
lar protein synthesis (Hargreaves 2013). The immobiliza-
tion of ammonia by heterotrophic bacteria depends on the
total carbon uptake by these bacteria and their gross con-
version efficiency. Ammonia immobilization and conver-
sion by heterotrophic bacteria are more rapid than
nitrification, because the growth and production of micro-
bial biomass per unit substrate of heterotrophic bacteria
are higher than that of nitrifying bacteria (Metcalf & Eddy
Inc. & Tchobanoglous 1991). However, when nitrifying
bacteria are stabilized, the addition of exogenous carbon
sources should be reduced or completely stopped to avoid
an excess of bioflocs production. Excessive supplementa-
tion of organic carbon (with high C/N ratio) can lead to a
rapid increase in settleable solids (SS), total suspended
solids (TSS), volatile suspended solids (VSS) and water tur-
bidity (Xu et al. 2016). Also, at high C/N ratio, autotrophic
biofloc is shifted to heterotrophic microbial biomass, lead-
ing to changes in water quality and biofloc composition
679
A. M. El-Sayed
(De Schryver et al. 2008). Increased biofloc accumulation
can also lead to a reduction in dissolved oxygen, an increase
in nitrogenous compounds, and may cause injuries to fish
gills (Ray et al. 2010b; Schveitzer et al. 2013b). Therefore, it
is necessary to keep the bioflocs suspended in the water and
in continuous contact with the nutrients and the oxygen,
through aeration. This would enhance the aggregation rate
and avoid biofloc precipitation. Therefore, quantitative and
qualitative application of carbon sources and aeration rate
in biofloc systems should be fully understood.
Carbon sources
Several studies have investigated the effects of different car-
bon sources and levels on the performance, health status
and water quality in biofloc-based shrimp aquaculture. The
effectiveness of carbohydrate sources in BFT systems
depends mainly on their carbon contents and degradation
speed, suggesting that some carbon sources are better in
promoting flocs production than others. In general, simple
sugars (such as molasses and dextrose) are degraded faster
than complex sugars (such as wheat bran), leading to lower
ammonia concentration, higher growth rates and better
water quality (Khanjani et al. 2017). Many natural products
and wastes have been used as carbon sources for BFT-based
shrimp farming, including sugarcane molasses, glucose,
sucrose, rice bran, wheat bran, starches and flour (see
Table 1 for review). Even biological degradation polymers
(BDPs), such as poly-caprolactone (PCL) and b-hydroxy-
butyric (PHB), have been tried, with varying results (Luo
et al. 2019). It should be emphasize, however, that the
effects of carbon addition depend on the carbon source,
quality and quantity, time and frequency of application,
total ammonia nitrogen (TAN) level in the water, supple-
mental feed and stocking density of farmed shrimps.
Molasses is among the most widely used carbon source,
during larval, nursery and grow-out phases for different
shrimps and prawns grown in BFT systems. Molasses addi-
tion to postlarval, nursery and grow-out BFT systems can
lead to significant decrease in TAN and nitrite-nitrogen,
and prevent build-up of these compounds (Samocha et al.
2007). Molasses also improves growth rates, survival rates
and water quality, and reduces the concentration of Vibrio
spp. Researches indicated that molasses are better than rice
bran and dextrose in reducing ammonia concentrations in
the culture water in BFT systems (Gao et al. 2012; Serra
et al. 2015; Huang et al. 2017).
The efficiency of a number of other carbon sources in
reducing ammonia concentration, improving water quality
and promoting microbial production and shrimp perfor-
mance has also been tested (Panigrahi et al. (2019a). Tested
sources showed varying efficiencies, with millets, molasses
and multigrain flour resulted in similar growth, survival
rate and disease resistance and were better than maida
680
flour, wheat flour, gram flour, rice flour and corn flour.
Tapioca and tapioca by-products were also highly efficient
in promoting shrimp survival and performance in BFT sys-
tems (Ekasari et al. 2014a).
C/N ratio
Not only carbon source determines the effectiveness of bio-
flocs, but also the ratio between carbon and nitrogen (C/N
ratio) in BFT system plays a crucial role in this process.
Maximum nitrogen uptake, waste decomposition and bio-
floc production generally occur at appropriate C/N ratio.
This means that the control of toxic inorganic nitrogen in
the water and the production of microbial flocs are
achieved by adjusting C/N ratios in feeds (Avnimelech
2015). On the other hand, if C/N ratio exceeds the mainte-
nance level, the concentration of TSS, VSS and SS will
increase, causing water turbidity, and may cause stressful
conditions to farmed animals (Xu et al. 2016).
Various C/N ratios have been used in BFT-based shrimp
culture, with varying, and sometimes contradictory, results
(See Table 1). For example, Xu et al. (2016) reared L. van-
namei juveniles (2.2 g) in a BFT system at C/N ratios of 9
(control, no carbon addition), 12, 15 and 18, using
molasses as a carbon source. They found that the values of
volatile suspended solids (VSS) and total reactive phospho-
rus increased, while total inorganic nitrogen declined, with
increasing C/N ratio. Better growth rates and FCR were
found at C/N ratio of 9 and 12 than at 15 and 18 treat-
ments. On the other hand, higher C/N ratios were reported
by other authors for optimal water quality and growth per-
formance of shrimp grown in BFT systems. Panigrahi et al.
(2018, 2019b) found that TAN, NO2-N and NO3-N levels
were significantly reduced at high C/N ratios (15 and 20)
compared to lower ratios (5 and 10). The best growth rates,
survival and FCR, and resistance against Vibriosis patho-
gens were also achieved at C/N ratio of 15–20. In general, a
C/N ratio of about 10–20 has been suggested for optimum
production of white shrimp L. vannamei, depending on the
carbon source used, growth stage, shrimp density and cul-
ture conditions.
Alkalinity and pH
As mentioned earlier (Section: types of biofloc systems),
water quality in BFT systems is controlled by natural pro-
cesses, including photosynthesis, nutrients uptake by algae,
oxidation of organic matter and nitrification–denitrifica-
tion reactions, in the presence of aeration and water mixing
(Hargreaves 2006). Ammonia is also regulated by phyto-
plankton uptake, nitrification and immobilization by bacte-
ria. In other words, plant photosynthesis and heterotrophic
and chemoautotrophic bacterial metabolism are responsi-
ble for different chemical transformations, causing water
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 Use of bioflocs in shrimp aquaculture

Table 1 A summary of the effects of different biofloc systems on water quality and zootechnical performance of shrimps and prawns

Species (size; g) System/ Carbon sources C/N ratio Performance remarks Ref
salinity

L. vannamei (2.68) Outdoor Molasses Heterotrophic bacteria effectively assimilated ammonia Ren et al.
tanks, SW and maintained good water quality. (2019)
Chemoautotrophic bacteria and photoperiod led to
the highest survival, performance, FCR and best
health status
L. vannamei (3.3) 850-L Molasses 12 Shrimp were exposed to 24L:0D, 12L:12D and 0L:24D. Baloi et al.
tanks, SW Light exposure resulted in higher production. (2013)
However, shrimp can be raised in total absence of
light with acceptable performance
L. vannamei (6.8) Indoor Molasses and wheat 16 Bioflocs enhanced growth performance and immune Zhao et al.
tanks, SW bran antioxidant parameters, especially at 1:0 and 1:1 (2016)
molasses: wheat bran ratio
L. vannamei (2.86) plastic bins, Molasses and cassava Biomass and survival were better at sugar cane Lobato et al.
BW flour molasses than cassava flour (2019)
L. vannamei (0.4) 3.8 m3 Sucrose 3.4 Natural light, metal halide lighting and/or relatively low Coyle et al.
tanks levels of fluorescent lighting (e.g. one light; 214 lx) (2011)
are suitable for indoor production of L. vannamei
grown in biofloc systems
P. vannamei (1.9- 15 m3 Molasses 10 Bioflocs significantly performance, survival and FCR Panigrahi
2.4) tanks, SW and played an immunostimulatory role in association et al. (2017)
with heterotrophic bacteria
L. vannamei (1) 500-L Molasses 5, 10, 15 TAN, NO2-N and NO3-N levels were reduced at C/N Panigrahi
tanks, SW 20 ratios 15 and 20. The best performance at C/N 15. et al. (2018)
Highest survival following challenge with Vibriosis was
obtained at C/N 10, 15 & 20. Higher expression of
immune genes was observed in BFT groups
Litopenaeus 1000-L Rice bran, molasses, 10–20 38–40% CP feed. Shrimp grown on BFT system Panigrahi
vannamei (PL) tanks ragi, wheat, branless allowed colonizing several beneficial bacteria in gut. A et al.
wheat and rice flour number of Bacillus strains produced all the (2019a)
extracellular enzymes along with antibiofilm activity
L. vannamei (1.48) Tanks, SW Molasses 15 40, 32 or 24% cp diets. Bioflocs improved Panigrahi
performance and FCR and showed higher resistance et al.
to disease, and higher immune genes stimulation, (2019d)
especially at 32 and 40% cp. Optimal dietary protein
could be reduced to 32% or lower
L. vannamei (1) 500-L Molasses 5, 10, 15, 35% cp diet. Performance, survival and immune (Panigrahi
tanks, SW 20 response were better in BFT-based treatment than in et al.
the control group. The best results were obtained at (2019b).
C/N 15
L. vannamei (0.42) 1.36 m3 No carbon source was 8.3 Bioflocs improved growth and FCR, reduced ammonia Ray et al.
tanks, added and pH levels and increased nitrite, nitrate and (2017)
turbidity
L. vannamei (4.8) 54-L tanks, Molasses 20 Growth, FCR, survival rates, energy retention and flesh Chan-Vivas
SW quality and acceptability were significantly better in et al. (2019)
BFT system than RAS
L. vannamei (3.11) Tanks Maize starch 15 IMTA with spotted scat and water spinach (Ipomoea Liu et al.
aquatica). The addition of starch into the IMTA system (2014)
improved growth, survival, productivity, profitability
and water quality
L. vannamei (1.3) Shaded Sucrose >10–16.92 75% and 100% of the theoretical adding quantity Gao et al.
tanks, SW reduced the concentrations of total ammonia (2012)
nitrogen and nitrite-nitrogen, and improved water
quality and growth rates

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© 2020 John Wiley & Sons Australia, Ltd 681
A. M. El-Sayed

Table 1 (continued)

Species (size; g) System/ Carbon sources C/N ratio Performance remarks Ref
salinity

L. vannamei (2.21) Shaded Molasses 12, 15, 18 Shrimp were fed a less expensive feed or a more Xu et al.
tanks, SW expensive feed. The best water quality, shrimp (2018)
performance and FCR were achieved with a C/N ratio
of 12:1 for both feeds
L. vannamei (6.95) Indoor sucrose 15, 20 Carbohydrate addition enhanced immune response Xu and Pan
tanks, SW and antioxidant status. Performance and digestive (2012)
enzyme activity in BFT were higher than in the control.
No significant difference between C:N 15 and 20
L. vannamei 500-L Molasses 9, 12, 15, Volatile suspended solids and total phosphorus Xu et al.
(2.2 g) tanks, SW 18 increased, while total inorganic nitrogen declined, (2016)
with increasing C/N ratio. Better growth rates and FCR
were found at C/N of 9 and 12 than at 15 and 18
L. vannamei (PL 800 and Molasses, dextrose and 15, Molasses and dextrose were better than rice bran in Serra et al.
3
and 4.1 g) 1200 m rice bran reduced reducing ammonia levels in nursery and grow-out (2015)
tanks to 6 phases, suggesting that simple sugars are degraded
faster than complex sugars
P. vannamei PL lined molasses, wheat and 15 Bioflocs lowered nitrogen and improved growth rates, Kumar et al.
(0.015) earthen sugar (8:1:1) survival and FCR. They also increased heterotrophic (2018)
ponds bacterial count and reduced total Vibrio count
P. vannamei (2 g) 100-L molasses, tapioca, 15 Highest growth reported in tapioca treatment, whereas Ekasari et al.
aquaria, tapioca by-product, tapioca by-product resulted in the highest survival. (2014a)
SW and rice bran BFT system had higher phenoloxidase activity,
respiratory burst activity, following a challenge with
myonecrosis virus (IMNV)
L. vannamei (4.8) Tanks, BW cane sugar 14.9 Integrated system. Four tilapia densities (0, 89, 178, Poli et al.
and Nile tilapia (19&) 265 fish m 3) at a shrimp density of 281 m 3. (2019a)
(9.6) Growth rates were not affected by stocking densities,
but the integrated system increased the yield by
31.2%
P. monodon 2.5 m3 tapioca powder 12 Substrates addition, but not stocking density, Arnold et al.
(PL15) tanks, SW significantly improved growth rates. Substrates also (2009)
(30.3&) reduced ammonia and nitrite concentrations,
especially at higher stocking density
F. paulensis (PL10- 40-L tanks, Molasses and wheat 20 Bioflocs with or without supplemental feed (40% CP). Emerenciano
25) SW bran The best growth rates were achieved in PL reared in et al. (2011)
biofloc in the presence of feed
L. stylirostris 30 m3 No carbon source added 8 BFT improved number of spawns per ablated female, Emerenciano
(broodstock) tanks, or number of consecutive maturations per female, et al. (2012)
earthen number of eggs per spawn, number of eggs per g bw
ponds and cumulative spawning rate
F. duorarum (12) 20 m3 molasses and wheat 20 BFT with fresh food (FF) had much higher egg Emerenciano
shaded bran production than those raised in conventional clear- et al. (2014)
outdoor water system
tanks
M. rosenbergii 50-L tanks, Brown sugar NA 32% cp diet. Growth and survival were similar in RAS Ballester
(0.13) FW and BFT systems, but FCR was better in RAS et al. (2017)
M. rosenbergii 20 m3 Molasses 20 Growth and body protein were higher, and FCR was P
erez-
(0.025) ponds, FW lower, in the BFT system than in traditional system Fuentes
et al. (2013)

SW, sea water; BW, brackish water; FW, fresh water; NA, not available.

quality changes (Hargreaves 2006). The pH and alkalinity At high stocking densities and feeding rates, system water
should be maintained at appropriate levels, 7-9 and 50– may turn acidic, due to increased nitrification. Nitrification
100 mg CaCO3 L 1, respectively (Avnimelech 2015). process leads to significant alkalinity consumption and

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682 © 2020 John Wiley & Sons Australia, Ltd

lowering of water pH. In this case, supplemental alkalinity
should be added to reduce the water acidity and, in turn,
increase the pH, alkalinity and total suspended solids (TSS)
in culture waters (Wasielesky et al. 2006; Avnimelech
2015). An alkalinity of about 100–150 mg CaCO3 L 1 or
Na2CO3 has been suggested for maintaining optimum
water pH and alkalinity, and also performance of biofloc
system (Furtado et al. 2015; Zhang et al. 2017). The con-
centrations of ammonia-nitrogen should also be kept
at < 1.5 mg L 1 in shrimp BFT system (Furtado et al.
2011), while nitrite-nitrogen concentration must
remain < 2 mg L 1.
Nonetheless, very few studies were carried out to evaluate
the effects of pH and alkalinity on water quality and shrimp
performance in BFT systems. The addition of sodium bicar-
bonate significantly increased water alkalinity in juvenile L.
vannamei tanks (Zhang et al. 2017). Adjusting the pH at
7.6–8.1 resulted in better water quality parameters, growth
rates and immune response than in treatment with non-ad-
justed pH. This finding suggested that a pH above 8.1 and
alkalinity >100 mg CaCO3 L 1 are required for optimum
shrimp growth, stability of the bioflocs and establishment of
nitrifying bacteria in BFT-reared L. vannamei. The same
conclusion was reached by Furtado et al. (2015) who found
that the highest ammonia and nitrite levels and lowest
growth rates of juvenile white shrimp (L. vannamei) in a
BFT system were reported at low alkalinity (75 mg CaCO3
L 1). Increasing alkalinity to 150 and 300 mg CaCO3 L 1
resulted in a significant improvement in these parameters.
Contrary to the above results, Pi
erri et al. (2015) found
that juvenile white shrimp (L. vannamei) (5.6 g) in BFT sys-
tem can be grown successfully at lower alkalinity. When the
shrimp were reared at different alkalinities (40, 80, 120 and
160 mg CaCO3 L 1), pH and TSS decreased at lower alkalin-
ity, while survival and growth rate were not significantly
affected. The authors concluded that L. vannamei can be suc-
cessfully grown in biofloc at 40 to 160 mg CaCO3 L 1. How-
ever, in this study, despite that the pH was lower at 40 mg
CaCO3 L 1 than at other alkalinity concentrations, it was
still close to 7 (neutral, not acidic), suggesting that supple-
mental alkalinity may have not been necessary. It should be
emphasized that maintaining pH and alkalinity in BFT-
based shrimp culture may be affected by the type of alkalin-
izing compounds used, as demonstrated by Furtado et al.
(2011). Calcium hydroxide (Ca(OH)2) and sodium bicar-
bonate (NaHCO3) were found more effective in supplement-
ing alkalinity than sodium carbonate (Na2CO3), despite that
sodium carbonate was effective in raising pH levels.
Aeration
In the nitrification process, ammonia is oxidized into
nitrite and subsequently to nitrate by ammonia-oxidizing
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Use of bioflocs in shrimp aquaculture
bacteria (AOB) and nitrite-oxidizing bacteria (NOB),
respectively (Ebeling et al. 2006). This process requires
sufficient and continuous oxygen supply; otherwise, it
will be suppressed (Avnimelech 2015). Therefore, oxy-
genation and homogeneous mixing of culture water are
essential for ammonia nitrification, bioflocs build-up,
and enhancing the activity of microbial communities
(Avnimelech 2015). On the other hand, excessive aeration
may reduce flocs aggregation and cause their rupture.
This means that aeration intensity should be maintained
at an appropriate rate to keep flocs in suspension with-
out causing their rupture, and ensure the nitrification
process (Lara et al. 2017a).
Since aeration intensities of BFT systems may affect per-
formance, water quality and biofloc composition, a number
of studies have addressed the effects of aeration systems
and intensities on shrimp culture in BFT systems. For
instance, diffused air systems (air blowers) resulted in bet-
ter formation of bioflocs and enhancement in the growth
performance of white shrimp (L. vannamei) than propellers
and vertical pumps (Lara et al. 2017a). Diffused air also
caused higher biofloc aggregation, whereas the use of verti-
cal pumps and propellers ruptured or broke apart these
bioflocs. Aerotubes are also better than air stones in water
movement and circulation, for biofloc formation in BFT-
based whiteleg shrimp culture (Harun et al. 2019). In the
meantime, the nitrification process is more efficient at a
high air flow rate in the presence bioflocs than at low rate
(de Morais et al. 2020). In addition, biofloc-based shrimp
tanks receiving stronger flow rate have better survival and
yield than at lower aeration intensity. However, intermit-
tent aeration strategy (e.g. 0.5-h aeration/0.5-h non-aera-
tion) may be more energy saving and, in turn, more cost-
effective than continuous aeration in BFT systems (Liang
et al. 2014).
Light exposure
Sunlight is the limiting factor for primary productivity and
phytoplankton blooming in eutrophic outdoor ponds,
because it enhances photosynthesis through the water col-
umn. Under photosynthetic conditions, algae provide oxy-
gen to the pond during daytime, which is consumed by the
algae and other microbial communities, leading to increas-
ing their mass production. Phytoplankton also have a
higher affinity for ammonia than bacteria; therefore,
ammonia is removed mainly by phytoplankton. At high
light intensity, phytoplankton density is sharply increased,
causing self-shading, which reduces light availability and
may cause phytoplankton crashes or die-out. This means
that photosynthesis leads to diel fluctuations in the concen-
trations of DO, CO2, pH and un-ionized ammonia (Harg-
reaves 2006; Avnimelech 2015). Under these conditions,
683
A. M. El-Sayed
the performance of biofloc systems remains under continu-
ous variations between day and night.
It is clear from the above discussion that the presence of
sunlight in BFT (especially in outdoor systems) is essential
for natural food production for fish and shrimp farmed in
biofloc systems (Ju et al. 2008). Light intensity and pho-
toperiod have been reported to affect shrimp performance
in both clear-water and BFT systems, depending on shrimp
species and developmental stages and also on whether the
practice is indoor or outdoor. Natural light enhances pro-
ductivity and availability of zooplankton in the system,
which are also used as natural food sources for farmed
shrimp. Meanwhile, higher light intensity is better than
lower intensity in promoting shrimp performance. At high
light intensity, growth rates and production of white
shrimp L. vannamei (Neal et al. 2010) and banana shrimp
(Penaeus merguiensis) (Hoang et al. 2003) were higher than
at lower intensity.
Light source and photoperiod
In addition to light intensity, light sources may also affect
the growth and survival of shrimp in BFT systems, as sug-
gested by Coyle et al. (2011). These authors reared L. van-
namei (0.4 g) in an intensive BFT system under natural
sunlight (12:12 light: dark cycle). Five light sources were
applied; one metal halide light, one fluorescent light, two
fluorescent lights and three fluorescent lights, with light
intensity of 718, 1074, 214, 428 and 642 lx, respectively.
Final biomass and survival were not significantly different
at natural light, metal halide lighting and one fluorescent
light. Increasing the number of fluorescent lights led to
blooming of filamentous bacteria, which causes gill fouling,
leading to lower shrimp survival. The results revealed that
natural light, metal halide lighting and/or relatively low
levels of fluorescent lighting (e.g. one light; 214 lx) are suit-
able for indoor production of L. vannamei grown in biofloc
systems. Light spectrum and intensity can affect bacterial
population structure, which has a profound effect on
shrimp survival and production.
In contrast with the effects of light intensity, photoperiod
may not have a significant effect on the performance of
shrimp grown in BFT systems. Baloi et al. (2013) demon-
strated that light may not be necessary for Pacific white
shrimp L. vannamei in BFT system, under different light
(L): Dark (D) regimes (24L:0D, 12L:12D and 0L:24D).
Despite that light exposure resulted in higher shrimp pro-
duction; Pacific white shrimp can be raised in total absence
of light with acceptable performance (Baloi et al. 2013).
Similar results were also found in white shrimp raised
under direct sunlight or at complete darkness (Esparza-Leal
et al. 2017). Shrimp survival growth rates, FCR and pro-
ductivity were not significantly different at both light
regimes.
684
Light and C/N management
As mentioned above, light is limiting for photosynthesis
and phytoplankton blooming, and in turn, ammonia
removal and floc build-up. The uptake of ammonia in bio-
floc systems occurs in heterotrophic bacterial assimilation
of ammonia-nitrogen to bacterial biomass. In this process,
light and C/N management are essential for the system
function. Carbon sources are also necessary to maintain the
optimum carbon/ nitrogen (C/N) ratio required for bacte-
rial assimilation (see Section: Carbon sources and carbon-
to-nitrogen ratio, for details). However, the combined
effect of light and C/N on ammonia uptake pathways is not
well understood. Only a couple studies have addressed this
issue, through investigating the combined effect of light
and C/N ratio on water quality, growth performance and
antioxidant enzyme activities in juvenile white shrimp L.
vannamei (Ren et al. 2019; Jiang et al. 2020).
Ren et al. (2019) reared white shrimp in heterotrophic
(biofloc) system with or without light, or in chemoau-
totrophic, no floc environment, where only feed was added
to the tanks. Chemoautotrophic treatments were more
dependent on light than heterotrophic treatments and had
more nitrite and nitrate accumulation. This has been attrib-
uted to the assimilation of ammonia by heterotrophic bac-
teria in the presence or absence of light. In addition, the
highest shrimp survival rate, feed efficiency and health sta-
tus were better in the chemoautotrophic (no floc) treat-
ment, with natural lighting (12h L: 12 h D) than in
heterotrophic environments. Along the same line, Jiang
et al. (2020) found that light and carbon source (molasses)
addition enhanced the microbial contents in heterotrophic
BFT systems used for L. vannamei culture.
Addition of substrate
Artificial substrates are commonly used in shrimp aquacul-
ture in BFT systems (Schveitzer et al. 2013b). These sub-
strates increase the surface area on which nitrifying bacteria
and other periphyton flocs are attached. These bioflocs can
be used as supplemental, natural food sources in BFT-based
aquaculture systems (Arnold et al. 2009; Avnimelech 2015).
Submerged substrates can also act as a safe refuge for
farmed shrimp, reducing competition for space and mini-
mizing aggressive behaviours, such as cannibalism, espe-
cially during moulting, leading to a significant reduction of
the negative impacts of high stocking densities (Arnold
et al. 2009; Audelo-Naranjo et al. 2012). Artificial substrates
can also improve water quality through reducing ammonia
concentration and stimulating nutrient recycling.
The addition of submersed substrates in shrimp culture
systems has been reported to improve water quality,
increase natural productivity, enhance periphyton forma-
tion, provide natural foods for farmed shrimp and improve
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© 2020 John Wiley & Sons Australia, Ltd

their growth rates and immune response (Arnold et al.
2009; Ferreira et al. 2016; de Morais et al. 2020). The effi-
ciency of substrates depends on substrate type and mode of
fixation, stocking density and aeration intensity. At high
stocking density, substrates addition, but not stocking den-
sity, significantly improved growth rates of Penaeus mon-
odon reared in a heterotrophic biofloc system (Arnold et al.
2009). Substrates also reduced ammonia and nitrite con-
centrations, especially at higher stocking density. The addi-
tion of substrates significantly reduced ammonia
concentration, stimulated nutrient recycling, enhanced nat-
ural food production and availability and increased growth
performance and yield of white shrimp L. vannamei in BFT
system (Audelo-Naranjo et al. 2012; Olier et al. 2020). It is
evident from these studies that the addition of substrates
can save dietary protein without adversely affecting growth
rates or body composition of L. vannamei reared in biofloc
systems.
Types and fixation format of substrate
Flocs settlement on submersed substrates may be affected
by the type and morphology of substrates used, presumably
due to the better ability of microbial adhesion on one sur-
face than on the other. The ability of substrates to trap TSS
may also vary from one source to another. However, only a
few studies evaluated the impact of substrate types and fixa-
tion format on shrimp farming in BFT systems. The
impacts of different types of artificial substrates on Pacific
white shrimp (PL20) reared in BFT system were evaluated
(Rezende et al. 2018). Substrate materials included Bidimâ
(geotextile), mosquito net screen (2 mm mesh) and Need-
lonaâ (polyester fibre), in addition to a control treatment
without substrate. Growth and survival to salinity stress
were not affected by the type of substrates, but mosquito
net screen and control treatments resulted in the highest
TSS values. The addition of Needlonaâ increased the sur-
vival rate and reduced TSS. These results indicate that
Needlonaâ is a suitable substrate for prenursery phase of
Pacific white shrimp reared in a biofloc system, because it
can efficiently maintain levels of TSS in culture water with-
out using clarifiers or water exchange, leading to higher
survival rates. Similar results were also reported on tiger
shrimp P. monodon juveniles (3.32 g) reared in BFT-based
system in the presence of bamboo mat and nylon mesh as
substrates (Kumar et al. 2019). The incorporation of nylon
mesh and bamboo mat in the BFT system trapped signifi-
cant proportion of TSS (31.3–38.6% and 8.5–13.5%,
respectively) and reduced bottom solid deposition. How-
ever, bamboo mats resulted in better development of bio-
film, higher microbial population, higher shrimp growth,
lower FCR and higher immune responses.
The effects of format and mode of artificial substrate fix-
ation on the performance of BFT-raised shrimp were
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Use of bioflocs in shrimp aquaculture
evident. Schveitzer et al. (2018) raised juvenile L. vannamei
(3.1 g) at high density in a BFT system applying different
substrate formats. Substrates were fixed vertically in the
tanks in three different configurations, completely fixed,
partially fixed, or frond format (screens more flexible),
while a treatment without substrates served as control. Sub-
strate-supplemented treatments provided similar growth
and FCR, but they were significantly better than the control
group. Survival rate was higher in the completely and par-
tially fixed substrates than in the frond format and the con-
trol groups. The authors suggest that in the frond format,
substrate screens become more flexible and more suscepti-
ble to water turbulence, leading to an adverse effect on
shrimp survival.
The addition of substrate in the presence of aeration in
biofloc systems may also play an integral role in promoting
biofloc production, water quality and shrimp performance.
High air flow enhances nitrification process, while the pres-
ence of substrates increases the surface area on which nitri-
fying bacteria are attached. The nitrification process,
shrimp (L. vannamei) survival and final biomass were bet-
ter at high air flow rate in the presence of substrates than in
clear-water tanks (Morais et al. 2020).
Total suspended solids
Total suspended solids (TSS) are an important factor
affecting biofloc production in intensive and super-inten-
sive aquaculture. Excessive supplemental organic carbon,
with high C/N ratio, can cause a rapid increase in TSS (Xu
et al. 2016). At high C/N ratio, autotrophic biofloc is also
shifted to heterotrophic microbial production, leading to
significant changes in water quality and biofloc composi-
tion (De Schryver et al. 2008). High concentration of TSS
(>500 mg L 1) may negatively affect shrimp health and
performance, especially at large sizes (>15 g) (Emerenciano
et al. 2012). This is because excessive particulate organic
matter can cause clogging of shrimp gills and decrease oxy-
gen exchange, leading to high mortality rates (Emerenciano
et al. 2012, 2013b). Excess flocs (400–800 mg L 1) may also
decrease DO concentration and, in turn, increase nitroge-
nous compounds (NH4-N, NO2-N and NO3-N) in aqua-
culture water (Vinatea et al. 2010).
It is clear that bioflocs system should be properly man-
aged by removing the excess TSS, which are removed
mainly by settling solids removal devices. TSS removal has
been reported to increase white shrimp (L. vannamei)
yields (Ray et al. 2010b). Initial TSS levels may also have
significant effects on water quality and shrimp perfor-
mance. When white shrimp were reared at different initial
TSS levels (low, medium and high; 100, 300 and 600 mg
L 1, respectively), settleable solids (SS), nitrite and nitrate
significantly increased with increasing initial TSS, whereas
685
A. M. El-Sayed

changes in alkalinity and pH revealed inverse correlations
(Gaona et al. 2017). At low initial TSS, water quality and
shrimp performance and survival were better than at med-
ium and high initial concentrations. In general, a range of
100–300 mg TSS L 1 was suggested to maintain successful
shrimp culture in BFT systems.
Nutritional value of biofloc for farmed shrimp
Biofloc, in general, have good nutrient profiles, making
them a potentially good natural food source for farmed
shrimp. Biofloc may contain 20%–>45% cp, <1%–>8%
lipids, <15%–>60% ash, <1%–>15% fibre and < 18%–
>35% total carbohydrates (Table 2). Accordingly,
composition, amino acid and fatty acid contents of bioflocs
vary greatly (Emerenciano et al. 2013b, 2014). The compo-
sition and nutritional value of biofloc depend on the car-
bon source, carbon-to-nitrogen (C/N) ratio of culture
water (Xu & Pan 2013, 2014a), dietary feed composition
and level (Xu & Pan 2014b), light intensity, composition
and age of biofloc aggregates, TSS, bacteria-phytoplankton
concentration, artificial substrates and the culture condi-
tions (Kuhn et al. 2008; Porchas-Cornejo et al. 2013; Avn-
imelech 2015). For instance, heterotrophic microorganisms
including bacteria, ciliates and flagellates generally bloom
at high concentration of dissolved organic carbon (Azam
et al. 1983). Also, biofloc communities included high con-
centrations of diatoms, copepods and polychaetes when

Table 2 Nutritional composition of biofloc materials from different system types

Species Composition (% dry weight) Remarks References

DM Cp Lipids Ash Fibre CHO†

L. vannamei 87.7 30.4 4.2 31.2 18.5 Khatoon et al.

(2016)
99.5–87.3 26.0–41.9 1.2–2.3 18.3–40.7 Farming systems affect the Ju et al. (2008)
results
12.0–13.9 28.8–43.2 2.1–3.6 22.1–42.2 8.7–10.4 Values were affected by Maic
a et al. (2012)
water salinity
49.0 1.1 13.4 12.6 36.4 Kuhn et al. (2009)
38.8–40.5 <0.1 11.8–24.7 15.3–16.2 Bioflocs from two different Kuhn et al. (2010)
fish effluents
21.3–32.1 1.6–2.8 43.4–61.4 Values varied according to Xu and Pan (2014a)
dietary protein and C/N ratio
29.3 2.0 33.2 7.4 28.1 Initial composition, before the Braga et al. (2015)
trial
18.6–23.2 0.6–1.4 47.9–54.3 1.5–2.7 24.8–25.7 Composition after the trial;
values affected by
composition of supplemental
food
27.3–31.6 3.7–4.2 43.7–49.4 Values varied according to the Xu and Pan (2012)
C/N ratio applied
16.2–19.2 26.4–29.5 0.8–1.1 27.9–43.3 8.6–16.7 Values affected by the types Lara et al. (2017a)
of aerators used (blower,
vertical pump, propeller)
F. brasiliensis 1.8–33.0 1.6–8.1 Values affected by biofloc size Magan ~a-Gallegos
classes; larger sizes (>50– et al. (2018)
<500 µm) have higher values
F. duorarum 24.7 0.6 47.3 2.4 26.3 Values are means of 6-month Emerenciano et al.
analyses (2013b)
28.0–30.4 0.5–0.6 35.8–39.6 3.1–3.2 18.1–22.7 Values affected by the Emerenciano et al.
presence of supplemental (2014)
fresh food
F. paulensis 30.4 0.5 39.2 0.8 2.9 (NFE)‡ Ballester et al.
(2010)
NA§ 38.3 0.4 31.6 16.6 19.0 Bioflocs produced from food Kuhn et al. (2016)
processing wastes

†Total carbohydrate.
‡Nitrogen free extract.
§Not applicable.

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686 © 2020 John Wiley & Sons Australia, Ltd

alfalfa enriched with molasses and vitamins were used as
substrates in shrimp earthen ponds (Porchas-Cornejo et al.
2013). When artificial substrates (Aquamats TM) were used
for intensive shrimp farming, periphyton consisted mainly
of primary producers (such as diatoms and cyanobacteria)
and primary consumers (such as rhizopods, heliozoans, cil-
iates, flagellates, foraminifera, copepods, rotifers and gas-
trotriches), in addition to detritivorous metazoans (such as
amphipods and nematodes) (Audelo-Naranjo et al. 2011).
Phytoplankton (mainly diatoms) may also dominate the
biofloc communities in outdoor cultures, under sufficient
sunlight and natural photoperiod, while in shaded or
indoor areas, with limited light or complete absence of
light, the heterotrophic bacteria may thrive and become the
dominant community (Becerra-Dorame et al. 2011; Godoy
et al. 2012).
As mentioned in the previous sections, farmed marine
shrimps and freshwater prawns can readily consume bio-
flocs as natural food sources. Cardona et al. (2015)
reported that natural food represented 37–40% of total
food consumption of shrimp L. stylirostris juveniles reared
in BFT system in the presence of supplemental feed. Folc
consumption can lead to significant improvements in
growth, feed efficiency, survival rate, digestive enzymes
activity, health status and water quality, in addition to a
significant reduction in feed cost (Emerenciano et al. 2011;
Cardona et al. 2015; Ray et al. 2019). The essential fatty acid
profiles of bioflocs are reasonably good (Toledo et al.
2016); therefore, they can help satisfy the lipid and fatty
acid requirement of farmed shrimps.
Full details or highlights on the use of bioflocs as natural
food for farmed shrimps during their different life stages
are provided in different sections of this review. Table 1
also summarizes the effects of bioflocs on zootechnical and
physiological responses, and water quality of different
shrimps farmed in BFT-based systems. Therefore, the rest
of this section will be allocated to the use of bioflocs as feed
ingredients or supplements, partially replacing the conven-
tional feed ingredients (e.g. fish meal and soybean meal) in
shrimp diets. The role of bioflocs in compensating the
reduction in dietary protein levels will also be highlighted.
Biofloc meal as a direct feed ingredient
Fish meal (FM) and soybean meal (SBM) have traditionally
been used as the main protein sources in aquafeed industry
worldwide (El-Sayed et al. 2000; El-Sayed 2020). The
increased demand for these sources, coupled with a signifi-
cant shortage in global supply (especially FM), has created
a sharp competition for their use by the animal feed indus-
try. This has led to a substantial increase in their prices in
recent years. Therefore, research for unconventional FM
and SBM alternatives for developing sustainable aquafeeds
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Use of bioflocs in shrimp aquaculture
has attracted the attention of many authors. Dried biofloc
has emerged as a potential feed ingredient, to partially
replace FM or SBM in aquafeeds.
As mentioned above, several factors can affect the nutri-
tional value of bioflocs and determine their potential as a
direct feed ingredient or a partial replacer of the conven-
tional diets. Factors such as nutrition priority, the ability of
farmed animals to ingest and digest microbial biofloc, and
biofloc composition, density and particle size should be
carefully analysed prior to the use of bioflocs as a feed
source (Ekasari et al. 2014b; Khanjani & Sharifinia 2020). It
has been found, for instance, that flocs with particle size of
>100–<48 µm are favourable for white shrimp L. van-
namei, due to their higher nutritional value (Ekasari et al.
2014b). The highest protein and lipid levels were found in
particle size of >100 µm, whereas flocs with a size
of <48 µm were rich in essential amino acids. Therefore,
varying supplemental levels of dietary biofloc have been
reported for optimum performance of farmed shrimps
(Khatoon et al. 2016; Anand et al. 2017; Ruby et al. 2017).
A relatively low replacement level (4–8% of the test diets)
was found to improve growth, survival and the activity of
digestive enzymes (amylase, cellulase, lipase and protease)
in juvenile Penaeus monodon (Anand et al. 2014) and
induce immunomodulatory effects and growth and physio-
logical enhancement (Anand et al. 2017). Meanwhile,
higher substitution levels were reported in other shrimp
species, as reported by Khatoon et al. (2016). These authors
evaluated the effects of replacing a commercial shrimp L.
vannamei postlarval diet with 0.0 (control), 25, 50, 75 and
100% dried waste biofloc. Biofloc samples were oven dried,
ground and milled to 300 µm and added to the tanks at the
proposed levels. The addition of bioflocs resulted in signifi-
cantly higher performance and survival than the commer-
cial diet, but the best performance was achieved at 50%
substitution level. On the other hand, a negative correlation
was found between the performance of L. vannamei and
dietary BFM levels (Ruby et al. 2017). However, BFM could
be considered as an alternative feed ingredient for white
shrimp due to their cost effectiveness. In the meantime,
more research is needed to further verify the use of BFM in
shrimp culture under different culture systems, feed com-
position, biofloc composition and shrimp developmental
phases.
Biofloc meal as a fish meal replacer
Biofloc meal (BFM) can not only partially replace the total
diet, but it can also replace the conventional dietary protein
sources. The partial replacement of convention protein
sources, such as FM and SBM, with BFM in shrimp diets
has received the attention of many researchers in recent
years. Studies have confirmed that the inclusion of BFM
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A. M. El-Sayed
can significantly reduce supplemental protein content of
the feeds, enhance growth performance of shrimp reared in
BFT systems and, in turn, reduce feed costs.
Microbial flocs, produced in sequencing batch reactors
(SBR) using sucrose as a carbon source, can replace up to
15.6% of SBM or FM protein in L. vannamei diets (Kuhn
et al. 2009). A higher substitution level (up to 30%) was
reported when bioflocs produced in SBR were used (Kuhn
et al. 2010). When bioflocs were produced in a membrane
biological reactor (MBR) without supplemental carbon,
flocs successfully replaced 21% of FM and SBM. At these
substitution levels, growth rates were higher than at bio-
floc-free diets. Similarly, bioflocs produced from food pro-
cessing wastes by ‘suspended-growth biological reactors’
successfully replaced up to 30% and 20% of dietary SBM
and FM, respectively, in diets for juvenile shrimp (species
not provided) (Kuhn et al. 2016). This result indicates that
bioflocs, harvested from food processing wastewater efflu-
ents, have a high potential as an alternative feed ingredient
in shrimp feeds. The use of these flocs can help reduce the
environmental impacts these wastes generally cause. How-
ever, at higher BFM inclusion levels, weight gain and nitro-
gen deposition were significantly reduced (Dantas et al.
2016; Gamboa-Delgado et al. 2017; Shao et al. 2017). It is
clear from the above results that 15% to 30% of conven-
tional protein sources can be replaced by BFM in shrimp
diets without negatively affecting their performance,
immune response and antioxidant status, but with a con-
siderable saving in feed cost.
Bioflocs can compensate for dietary protein reduction
The protein requirements of shrimp grown in BFT systems
can be significantly reduced, since they consume bioflocs as
a natural food, compensating for the reduction in dietary
protein. This assumption has been tested and validated by
several authors, using different shrimp species and sizes
and experimental designs (e.g. with or without clear-water
controls) (Table 3). Varying results have been reported,
due to the variation in biofloc age, composition and bio-
mass, diets composition, shrimp developmental stage,
water quality and experimental periods. For example, Pani-
grahi et al. (2019d) fed Juvenile L. vannamei (1.48 g) diets
containing 24, 32 or 40% cp for 130 days, in a BFT-based,
heterotrophic system. Shrimp raised in BFT system had sig-
nificantly better growth, survival, feed utilization, health
status and higher immune genes expression than the con-
trol group (autotrophic condition + 40% cp diet). They
also exhibited higher resistance to disease when challenged
with Vibrio parahaemolyticus pathogens. This finding sug-
gests that optimal dietary protein requirement of white
shrimp in a biofloc system can be reduced by 20% (from
40% to 32%). About 25% reduction in protein requirement
688
of postlarval L. vannamei (PL10) nursed in a BFT has also
been reported (Correia et al. 2014). Similar results were
also found with white shrimp (P. vannamei) and Indian
white shrimp (P. indicus) reared in a heterotrophic BFT
system (Panigrahi et al. 2020b), where a 30% cp diet pro-
duced higher performances and better water quality than
higher protein levels. The above results demonstrate that
over 25% of dietary protein can be saved when shrimps
were reared in BFT systems, leading to significant economic
benefits.
Dietary energy may also have a significant effect on dietary
protein requirement of BFT-raised shrimp. At appropriate
protein: energy (P: E) ratios, dietary protein can be spared
for growth at the expense of dietary energy (El-Sayed &
Teshima 1992; El-Sayed & Kawanna 2008). This assumption
was tested in juvenile whiteleg shrimp (0.97 g) reared in a
heterotrophic biofloc system and fed test diets containing
two protein (30 and 35%) and three digestible energy (DE)
(16, 17.5 and 19 kJ DE g 1) levels (Hamidoghli et al. 2018).
A diet containing 35% cp and 17.5 kJ g 1 energy (DE) was
suggested for optimum performance. However, in this par-
ticular study, a different conclusion may have been reached
if a wider dietary protein range had been tested, since only
two protein levels were used.
Not only Penaeid shrimp can benefit from biofloc as a
food source and save supplemental protein, but also fresh-
water prawns were found to consume biofloc as a partial
dietary protein replacer. When juvenile prawn Farfantepe-
naeus paulensis (72 mg) were fed test diets containing
graded levels of dietary protein (25, 30, 35, 40 and 45% cp)
in suspended microbial flocs, the best growth rates and
FCR were attained at 35% cp (Ballester et al. 2010). This
finding confirms the above results and indicates that micro-
bial flocs in BFT systems can limit the dependence on high-
protein diets and, in turn, reduce the production costs.
However, many of the above studies were carried without
clear-water, non-biofloc treatments, as controls (for com-
parison). Without such comparisons, it would be difficult
to assure whether the presence of biofloc has reduced pro-
tein requirement in these trials.
Role of biofloc in lipid nutrition of BFT-raised shrimp
As with protein requirement, lipid levels of shrimp raised
in BFT systems can be reduced, assuming that biofloc can
compensate for the reduced lipid levels. However, only a
few studies have been carried out to support this assump-
tion. For example, when juvenile L. vannamei (2.87 g) cul-
tured in heterotrophic BFT system and fed diets containing
different lipid levels (8.5, 9.5 and 10.5%), the highest sur-
vival was achieved at 8.5% lipid (Toledo et al. 2016). On
the other hand, growth rates, FCR and total haemocytes
count (THC) were similar among all treatment, suggesting
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 Use of bioflocs in shrimp aquaculture

Table 3 The use of bioflocs as supplemental feed/feed ingredients for farmed shrimps

Species and size (g) Levels tested/replaced Levels Remarks References

recommended

L. vannamei (PL 13) BFM†:10–30% of FM‡ 30 Flocs were produced in sequencing batch reactors, Kuhn et al.
and SBM§ sucrose used as a carbon source. Growth was higher (2010)
than the control
BFM: 10–21% of FM and 21 Flocs were produced in a membrane biological reactor
SBM without carbon addition. Growth was higher than the
control
L. vannamei (23 mg) BFM: 7.5, 15 & 30% of 30 30% BFM resulted in the best growth rates and PER Dantas et al.
FM (2016).
L. vannamei (6.5) 20, 25, 30 & 35% cp 25 Performance and health status were not significantly Xu and Pan
different, except at 20%. Dietary protein requirements (2014b)
can be reduced from 35% to 25%
F. paulensis (72 mg) 25, 30, 35, 40 & 45% cp 35 Shrimp fed diets with 35% or higher attained better Ballester et al.
performance (2010)
L. vannamei (1.48) 24, 32 or 40% cp 32 Shrimp raised in BFT system had better performance and Panigrahi et al.
health status than the control group (2019d).
L. vannamei (PL10) 30 and 40% cp 30 Growth rate was higher at 40% cp than at 30% cp; but Correia et al.
survival and PER were not significantly different 2014)
L. vannamei (0.97) 35 and 40% cp (at 16, 35% cp and 17.5 kJ g 1 DE could be considered as an Hamidoghli et al.
17.5 and 19 kJ g 1 DE optimal P:DE ratio for shrimp reared in a BFT system (2018)
L. vannamei (0.59) BFM: 25, 50, 75 and 25 Growth was negatively correlated with BFM. The growth Gamboa-
100% of FM at 25% BFM was comparable to all FM diet Delgado et al.
(2017)
P. vannamei and 25, 30 and 35% cp. 30 Growth and FCR in both species were highest at 30% Panigrahi et al.
P. indicus cp. Bioflocs improved the innate immunity in the two (2020b)
species
L. vannamei (4.1) FM: 0, 10, 20 and 30% 0.0 FM was replaced by a vegetable protein mixture. FM Moreno-Arias
could be excluded in BFT system, without adverse et al. (2018)
effects
L. vannamei (0.9) SBM: 0, 33, 67, & 100% 33% of dietary FM can be replaced by SBM, even Yun et al. (2017)
of FM without supplementation of methionine and lysine

†Biofloc meal.
‡Fish meal.
§Soybean meal.

that 8.8% dietary lipid is sufficient for L. vannamei reared
in BFT system. However, it should be mentioned here that
the range of dietary lipid levels tested in this study is nar-
row (8.5–10.5%; only 2% range). Lower lipid requirement
may have been achieved if a wider range had been used, as
demonstrated by Hamidoghli et al. (2020). These authors
fed juvenile L. vannamei (0.95 g) in BFT system at a wider
range of dietary lipids (4.5, 6.0, 9.0, 12.0 and 15.0%). About
5.6 to < 6.0% dietary lipids were found sufficient for opti-
mum performance. However, these values are much lower
than that reported by other authors (Zhang et al. 2013; Xie
et al. 2019), where the lipid requirements of L. vannamei in
traditional systems range from 10–12.4%. These findings
indicate that dietary lipids can be significantly reduced in
BFT-raised shrimps compared to those reared in tradi-
tional, clear-water systems.
Once again, the above results may be questionable,
because no clear-water treatments were used as control
trials. In addition, lipid quality and sources, dietary energy,
shrimp species, sizes and genetic lines and biofloc composi-
tion may have affected the above results. For instance,
Emerenciano et al. (2014) reported that both n-3 and n-6
essential fatty acids (EFA) in bioflocs vary considerably,
which, in turn, may affect the shrimp performance. Also,
bioflocs rich in lipid and protein have been suggested to be
utilized efficiently for the initial stages of gonad develop-
ment of farmed shrimp (Avnimelech 2015).
Role of biofloc in compensatory growth
Shrimps are characterized by compensatory growth after
short periods of feed restriction or fasting (Lin et al. 2008;
Zhu et al. 2016). Therefore, cycles of shrimp starvation and
re-feeding may improve their performance (Dong & Wu
2001). Thus, in the presence of bioflocs, shrimp feeding
could be reduced through feed restriction or short-fasting

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© 2020 John Wiley & Sons Australia, Ltd 689
A. M. El-Sayed
periods, since the animals can use bioflocs as a compen-
satory food source.
This approach was analysed by a couple of authors. Kaya
et al. (2019) subjected BFT-reared speckled shrimp Metape-
naeus monoceros to gradual decreasing of feeding levels
from 4% to 1% body weight (bw) per day or exposed them
to 2 days of feeding with 4% bw, followed by 1 day fasting,
or 1-day feeding and 1-day fasting. The growth rates, feed
efficiency, survival, immune system and whole body com-
position in BFT groups were better than in shrimp reared
in traditional system, using the same experimental design.
Biofloc consumption also increased with decreasing feeding
levels and adopting feeding restrictions. Similar results were
obtained by Lara et al. (2017b) on L. vannamei juveniles
(0.89 g) fed a test diet at a daily rate of 0, 0.6, 1.2 and 1.8%
bw in the presence or absence heterotrophic flocs. A 1.2%
feeding level, in the presence of microbial biofloc, was suffi-
cient for optimum shrimp performance. At this feeding
level, 35% reduction of the total feed consumption could
be achieved.
Role of biofloc as a probiotic in shrimp culture
In addition to their benefits highlighted in the previous sec-
tions, bioflocs can also serve as probiotics, which can
improve the immune systems and prevent disease out-
breaks and also stimulate digestive enzyme activities. This
has been attributed to the ability of bioflocs to maintain
good water quality and promote the production of bacteria
that produce short-chain fatty acids, which can protect the
intestinal epithelial cells and prevent diseases. Microbial
flocs also disrupt the quorum sensing of pathogenic bacte-
ria (Avnimelech 2015), leading to clear probiotic activities
(Emerenciano et al. 2013a; Adel et al. 2017). Several studies
have been published on the use of bioflocs as antibiotics,
antifungals and immune system stimulator for preventing,
controlling and reducing pathogens in BFT-raised shrimps.
Immunostimulatory effects
The possible immunostimulatory effects of bioflocs on
farmed shrimps have been widely documented. Bioflocs
can enhance innate, non-specific, immune system of cul-
tured shrimps through providing a wide range immunos-
timulants against infections. For example, microbial cell
walls in the bioflocs contain lipopolysaccharides, glucans
and peptidoglycans. These compounds can activate the
immune response in shrimps through enhancing the non-
specific immune mechanisms, leading to significant
enhancement in the immune system in farmed shrimp
(Aguilera-Rivera et al. 2019a; Panigrahi et al. 2019c,
2020a). Therefore, BFT-reared shrimp showed higher
resistance to disease after challenge with Vibrio
690
parahaemolyticus. Consequently, immune response param-
eters, such as total hemocyte count (THC) and prophe-
noloxidase (ProPO) activity, were significantly increased in
heterotrophic biofloc-reared white shrimp, compared to
non-biofloc trials (Panigrahi et al. 2019a). Bioflocs are also
capable of accumulating a bacterial compound poly-b-hy-
droxybutyrate (PHB), which has been shown to improve
growth and food digestibility and protect farmed aquatic
animals from bacterial infections (Kuhn et al. 2009; De
Schryver et al. 2010; Khanjani & Sharifinia 2020). In addi-
tion, microbial probiotics include viable microbial cells,
which can enhance growth, intestinal digestive enzymes,
feed digestion and absorption, and immune response, in
addition to inhibiting pathogenic microorganisms (Pani-
grahi et al. 2017; Panigrahi et al. 2020a). Therefore, shrimp
grown in BFT environments exhibit better health status
and growth performance (Kuhn et al. 2009).
Bioflocs contain a high number of heterotrophic, benefi-
cial microbial communities, including Bacillus, Sphin-
gomonas, Pseudomonas, Micrococcus, Nitrospira, Nitrobacter
and yeast. These microorganisms can be used as probiotics
in aquaculture, leading to improving water quality, perfor-
mance and health of cultured aquatic animals (Monroy-
Dosta et al. 2013; Adel et al. 2017). For example, Bacillus,
gram-positive, rod-shaped beneficial bacteria, are among
the most dominant groups in the microbial communities
in bioflocs (Panigrahi et al. 2019c), which may play an
important role as a probiotic in shrimp farming. The pres-
ence of Bacillus can lead to several benefits to farmed
shrimp, including improving growth rates, survival, feed
efficiency, immunity and disease resistance (Adel et al.
2017; Pacheco-Vega et al. 2018; Panigrahi et al. 2020c).
Bacillus-contained bioflocs were found to improve immu-
nity in Indian shrimps P. indicus (Panigrahi et al. 2020c).
Panigrahi et al. (2019c) found also that more beneficial
bacteria were colonized in the gut of white shrimp grown
on heterotrophic bioflocs than those reared in a control,
clear-water system. Bacillus were also found to produce dif-
ferent extracellular enzymes, providing potential benefits in
terms of growth and feed digestion, absorption and meta-
bolism.
Pathogenic bacteria, such as Vibrio, Aeromonas and Pseu-
domonas, can heavily accumulate in BFT-based systems and
cause pathogenicity through increasing or decreasing the
concentrations of suspended solids (Khanjani & Sharifinia
2020). Bacterial diseases, such as Early Mortality Syndrome
(EMS), (currently known as Acute Hepatopancreatic
Necrosis Disease (AHPND)) caused by Vibrio para-
haemolyticus, have also been reported in shrimp farms in
different parts of the world (Avnimelech 2015). A vibriosis
outbreak was recorded in the Pacific white shrimp, L. van-
namei reared in heterotrophic biofloc and clear-water sys-
tems in Mexico in 2015 (Aguilera-Rivera et al. 2019b).
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The presence of ripe biofloc communities can reduce the
prevalence and growth of pathogens in shrimp ponds by
competing for nutrients, space and substrates (Emeren-
ciano et al. 2013a). Bioflocs also produce inhibiting com-
pounds that reduce the number and growth of these
pathogens. For example, polymer products, such as poly-b-
hydroxybutyrate (PHB), are produced by many biofloc
microorganisms, acting as a preventive or curative agent
against Vibrio infections (Avnimelech 2015). When white
shrimp were fed Bcillus-inoculated bioflocs, or reared in
Bacillus-containing environment, the Vibrio load was sig-
nificantly reduced (Liu et al. 2017; Pacheco-Vega et al.
2018; Khanjani & Sharifinia 2020). In addition, when white
shrimp were reared in biofloc environment and then chal-
lenged with pathogenic V. Harvey, the immune system was
able to eliminate the pathogen and avoid disease occur-
rence (Aguilera-Rivera et al. 2014). Biofloc production has
also been reported to reduce the concentration of Vibrio
spp. in pink shrimp (Farfantepenaeus brasiliensis) (de Souza
et al. 2014) and white shrimp (L. vannamei) culture waters
(do Esp
ırito Santo et al. 2017). In addition, when L. van-
namei were grown in BFT-based, heterotrophic system,
they had significantly better growth, survival, feed utiliza-
tion, health status and higher immune genes expression
than shrimp reared in autotrophic conditions (Panigrahi
et al. 2019d). BFT-raised shrimp also exhibited higher resis-
tance to disease when challenged with V. parahaemolyticus.
It is clear from these studies that Bacillus strains such as
Lactobcillus plantarum and L. lactis, which are a major com-
ponent of bioflocs, could be successfully used as a protec-
tive agent against Vibrio infection.
Antioxidant effects
Bioflocs can enhance antioxidant components in farmed
shrimps and prawns and, in turn, stimulate their immune
responses (Xu & Pan 2013; Anand et al. 2017). The expres-
sions of hemocytes enzymes related to antioxidants status
were enhanced in white shrimp reared in BFT system (Xu
& Pan 2013). Also, the expression of genes related to
antioxidant defence systems (SOD, MnSOD, CAT) were
significantly upregulated in white shrimp grown in BFT
environment (Panigrahi et al. 2019a), leading to an
enhancement in their immune system. The expression of
six selected genes related to innate immunity, namely
ProPO1 and ProPO2, serine protease (SP1), prophenoloxi-
dase activating enzyme (PPAE1), masquerade-like serine
protease (MAS) and Rat-sarcoma-related nuclear protein,
was significantly upregulated in BFT-grown shrimp, com-
pared to shrimp reared in no-BFT system (Kim et al. 2014).
Biofloc microbial communities can also affect the antiox-
idant and oxidative damage responses in shrimp grown in
BFT system. For instance, glutathione (GSH) concentration
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© 2020 John Wiley & Sons Australia, Ltd
Use of bioflocs in shrimp aquaculture
in the gills and hepatopancreas was lower in BFT-raised L.
vannamei than in those reared in traditional, seawater sys-
tem (da Silva Martins et al. 2015). Glutathione peroxidase
was upregulated only in BFT group; superoxide dismutase
(SOD) expression was lower in BFT-reared shrimp than
those grown in recirculating system without biofloc (Soto-
Alcal
a et al. 2019). In addition, the catalase activity
increased in BFT-grown shrimp (Cardona et al. 2015).
These findings suggest that flocs can play a significant role
in immune stimulation and in modulating the transcrip-
tion of immune-related genes and, therefore, improve the
shrimp immune response and prevent the development of
disease infection.
The effects of bioflocs on the immunity response of
another shrimp species, L. stylirostris broodstock reared in
BFT system, were also evident (Cardona et al. 2016).
Higher GSH level and total antioxidant status (TAS), and
lower oxidized/reduced glutathione ratio were observed in
broodstock reared in BFT system than in those reared in
clear-water system. Also, the biofloc treatments improved
the innate immunity of Indian white shrimp (P. indicus)
(Panigrahi et al. 2020b). Significant effects of bioflocs on
the growth, immune and antioxidant response of other
commercially important farmed species, including black
tiger (P. monodon) (Anand et al. 2014, 2017; Kumar et al.
2017) and L. stylirostris (Cardona et al. 2016) have also been
documented. Similar results were also reported in freshwa-
ter prawn (Macrobrachium rosenbergii) (Miao et al. 2017)
and Chinese shrimp (F. chinensis) (Kim et al. 2015) where
augmentation of biofloc medium with Bacillus improved
immune response and reduced oxidative stress.
Application of bioflocs in shrimp culture
The vast majority of published researches on rearing
shrimp in BFT systems focussed on the growth perfor-
mance, feed utilization, survival and production potentials,
in addition to water quality improvement (Table 1). The
effects of bioflocs on growth promotion depend on shrimp
species and developmental stage, feeding traits and floc
density (Crab et al. 2010; Avnimelech 2015). Most of these
studies indicated that bioflocs significantly increased
growth rates of penaeid shrimp and freshwater prawn dur-
ing larval, nursery and grow-out phases, in semi-intensive,
intensive and supper intensive farming systems (de Lorenzo
et al. 2016; Ren et al. 2019; Lobato et al. 2019). As men-
tioned above, shrimps can readily consume bioflocs as an
additional protein source, resulting in a significant
improvement in shrimp performance and meat quality and
a reduction in production inputs. This section is dedicated
to the application of BFT in shrimp culture at different
developmental stages, namely larval, nursery, juveniles and
grow-out stages and broodstock.
691
A. M. El-Sayed
Larval stages
Hatchery development and larval rearing is a bottle neck
for successful shrimp aquaculture industry. In this regard,
several papers have been published on the impact of bio-
flocs on larval performance, survival and immune response,
especially during the past few years (Table 1). The effects of
bioflocs on larval performance may depend on shrimp spe-
cies. For example, Kim et al. (2015) evaluated the effects of
bioflocs as natural food sources for the postlarvae (PL10-
PL20) of three Penaeid species (L. vannamei, F. chinensis
and M. japonicus), in the presence or absence of supple-
mental feeds (45% cp). The presence of both biofloc and
feeds resulted in better growth rate of L. vannamei PL than
in biofloc-based treatment, whereas the growth and sur-
vival rates of the other two species were not affected by
dietary treatments. These results suggest that L. vannamei
imply filter-feeding habits (Gelabert & Pacheco 2011),
while F. chinensis are cannibalistic, as supported by Li et al.
(2019) who reported frequent cannibalism of F. chinensis.
The feeding mechanism of M. japonicus was not fully
understood.
Heterotrophic biofloc communities have been reported to
significantly improve the growth, survival rates and FCR of
shrimp postlarvae (Emerenciano et al. 2012; Kumar et al.
2018). Bioflocs can also improve feed digestibility and
absorption in shrimp larvae, by improving colonization of
beneficial bacteria in the guts of cultured shrimp. They also
enhance extracellular enzyme production such as amylase,
protease, lipase, cellulase, xylanase and pectinase in white
shrimp, through colonization of beneficial bacteria in the
guts of shrimp grown on floc communities than in clear
water (Panigrahi et al. 2019c). The wide diversity of biofloc
microorganisms and their high contents of protein, amino
acids and fatty acids may have promoting effects on larval
performance and feed digestion and absorption. Bioflocs can
also sharply reduce total ammonia nitrogen (TAN) and
nitrite (NO2–N) concentrations in BFT ponds, concomitant
with a significant increase in heterotrophic bacterial count
and reduction in total Vibrio count (Kumar et al. 2018).
Nursery stages
Nursery phase has been given considerable attention in
shrimp aquaculture, because it is a critical transitional stage
between early larval stage and grow-out stage. Appropriate
management during this phase can lead to rapid growth of
the cultured species (Emerenciano et al. 2012) and can
allow much higher stocking densities (Wasielesky et al.
2013; Panigrahi et al. 2020a). However, high stocking den-
sity in the nursery phase may reduce shrimp survival,
because of their cannibalistic behaviour (Abdussamad &
Thampy 1994).
692
BFT has been applied successfully in nursing different
shrimp and prawn species, including L. vannamei (Esparza-
Leal et al. 2015; Arias-Moscoso et al. 2018; Panigrahi et al.
2020a), P. monodon (Arnold et al. 2009), F. paulensis
(Emerenciano et al. 2011), F. brasiliensis (de Souza et al.
2012; Emerenciano et al. 2012), F. setiferus (Emerenciano
et al. 2009) and F. duorarum (Emerenciano et al. 2013a)
and freshwater prawn M. Rosenbergii (P
erez-Fuentes et al.
2013; Ballester et al. 2017). These species have been
reported to readily consume bioflocs as a natural food
source, leading to significant improvements in growth, feed
efficiency and survival rate, in addition to a reduction in
supplemental feeds. Bioflocs can even replace commercial
feed completely or partially, in addition to improving water
quality, and enhance health status of nursed shrimps.
The presence of diatom-inoculated bioflocs led to over
60% increase in the final weight, and about 50% in survival
of nursed F. paulensis postlarvae, compared to clear-water
system (Emerenciano et al. 2011). When supplemental feed
was offered, in addition to bioflocs, the weight gain was
almost doubled, while survival rate increased by 170%. The
same authors (Emerenciano et al. 2012) reported also that
growth and survival rates of F. paulensis postlarvae reared
in heterotrophic flocs system were significantly higher than
in the clear-water system. However, supplemental feed to
bioflocs did not improve the performance and survival
rates any further, indicating that supplemental feed may
not be necessary during nursery phase of this species. When
P. vannamei PL-3 were reared at high stocking densities in
BFT system, they showed extremely higher growth rates,
survival rate, heterotrophic bacterial counts (and lower
Vibrio load) and better water quality than in BFT-free
group (Panigrahi et al. 2020a). BFT also upregulated cer-
tain immune genes, such as prophenoloxidase, crustin,
MAS, serine protease, alpha-2-M and antiviral genes.
Juvenile and grow-out phases
Biofloc technology can play a significant role in the opti-
mization of growth rates, survival, feed efficiency, health
status, meat quality and net profitability of different shrimp
species during the juvenile and grow-out phases (Wasie-
lesky et al. 2013; Yun et al. 2017; Panigrahi et al. 2017). Bio-
flocs can successfully be used as a natural food or
supplemental ingredients, especially for marine white
shrimp Litopenaeus vannamei (Wasielesky et al. 2013; Yun
et al. 2017; Panigrahi et al. 2018).
Studies indicated that growth performance, survival and
feed utilization efficiency of juvenile and adult white
shrimp grown on heterotrophic biofloc without supple-
mental feed were superior to those reared in a clear-water
environment and fed high or low protein diets (Wasielesky
et al. 2006). When juvenile white shrimp (5.3 g) were
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raised in biofloc-dominated water and fed 35 or 40% cp
diet, their performance was not significantly different
between the two feed types (Prangnell et al. 2016). How-
ever, the 40% cp feed caused higher nitrate and phosphate
concentrations in culture water and enhanced alkalinity
consumption and bicarbonate use, leading to higher phyto-
plankton density. Similar results were found by Yun et al.
(2017), where the presence of biofloc can save about 10%
of dietary protein without adversely affecting growth rates
of L. vannamei. These findings reveal a significant contribu-
tion of microbial biofloc as natural food to L. vannamei
performance. The results also suggest that low protein feeds
can be used in BFT systems.
The biofloc and periphyton technology (BPT) is a biofloc
system to which substrates and carbon source are added.
The application of BPT can improve growth and
immunomodulatory performance of Pacific white shrimp
P. vannamei during nursery and grow-out culture (Pani-
grahi et al. 2017). Shrimps grown in BPT system attained
significant improvement in growth rate, biomass at harvest,
survival and FCR than in the control, clean-water treat-
ment. In addition, the non-specific immune response in
BPT-reared shrimp was enhanced, suggesting that bioflocs
can play immunostimulatory roles in association with het-
erotrophic bacteria.
Promising results have also been reported on commercial
culture of freshwater prawns in BFT-based systems (Crab
et al. 2010; P
erez-Fuentes et al. 2013; P
erez-Rostro et al.
2014). Bioflocs provided significant nutritional contribu-
tion to pond-reared M. rosenbergii (P
erez-Fuentes et al.
2013). The surface of bioflocs increased the surface area on
which the bacteria adhere, enhancing biofloc production
and allowing higher stocking density and better survival of
Malaysian prawn (M. rosenbergii) (P
erez-Rostro et al.
2014). Another promising candidate for aquaculture in
BFT systems is the Chilean river shrimp (Cryphiops caemen-
tarius). This species can tolerate a wide range of environ-
mental conditions, such as temperature, pH, hardness,
alkalinity, suspended solids, ammonia, nitrites and nitrates
(Ulloa Walker et al. 2020). Preliminary results indicated
that C. caementarius grown in heterotrophic BFT environ-
ment had higher growth and survival rates than in clear-
water system (Ulloa Walker et al. 2020).
Shrimp broodstock
The bioflocs can improve spawning performance and larval
survival of shrimp broodstock, as demonstrated by
Emerenciano et al. (2011, 2013c, d, 2014). Spawning per-
formance (latency period, number of spawns per ablated
female, number of consecutive maturations per female,
number of eggs per spawn, number of eggs per g bw, and
cumulative spawning rate) of blue shrimp L. stylirostris
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Use of bioflocs in shrimp aquaculture
broodstock fed high-protein diet and fresh food (FF) was
significantly higher in floc-based spawners than in those
reared in the earthen ponds (Emerenciano et al. 2013c).
The eggs of shrimp females fed on bioflocs and FF con-
tained higher levels of total n-3 and n-6 HUFA than those
reared on bioflocs only (Emerenciano et al. 2013d). In
addition, feed supplementation resulted in higher biofloc
crude protein, lipid, fibre and ash content than biofloc
tanks without FF.
Similar results were also reported on shrimp (L. styliros-
tris) (Cardona et al. 2016) and pink shrimp F. duorarum
broodstock (Emerenciano et al. 2014). BFT-reared brood-
stock showed better final survival rate and health status,
higher gonado-somatic index, higher spawning rate and
frequency, higher number of spawned eggs and higher lar-
val survival than broodstock raised in clear-water systems.
Bioflocs also significantly enhanced the immunity response
of L. stylirostris broodstock (Cardona et al. 2016). Higher
GSH level and total antioxidant status (TAS), and lower
oxidized/reduced glutathione ratio were observed in brood-
stock reared in BFT system than in those in clear-water
autotrophic system. The contribution of natural food from
microbial flocs to spawning performance of another species
(F. brasiliensis) was also evident (Maga~ na-Gallegos et al.
2018).
The above sections clearly demonstrate the role that nat-
ural productivity in BFT system can play in shrimp nutri-
tion and production management during larval, nursery,
juvenile, grow-out and broodstock phases. The competi-
tiveness of BFT system is not only economic, but also eco-
logical. Bioflocs are an ideal alternative for traditional,
water-demanding systems, especially in areas characterized
by harsh climate and water scarcity. They can also reduce
the volume of water required for traditional farming sys-
tems. In addition, they reduce the amount of wastes that
would, otherwise, be dumped into the aquatic environ-
ment, through transferring these wastes into natural foods
for farmed species.
Negative impacts of bioflocs on shrimp
performance
Despite the significant benefits of bioflocs in shrimp and
prawn aquaculture, these crustaceans may not benefit from
bioflocs if BFT systems are not properly understood and
managed. A number of studies revealed that shrimp perfor-
mance and water quality were better in recirculating aqua-
culture systems (RAS) than in BFT systems. For example,
ammonia concentrations in the tanks of freshwater prawn
Macrobrachium rosenbergii PL grown in BFT system
exceeded the safe levels, compared to RAS with biofilters
(Ballester et al. 2017). Growth and survival rates were also
similar, whereas FCR was better in RAS treatment than in
693
A. M. El-Sayed
floc-based treatment. The poor performance in floc-based
trial may have been due to elevated ammonia concentra-
tion (2.77 mg L 1), which may have caused oxidative stress
and reduced shrimp performance (Pinto et al. 2016). Ray
and Lotz (2017) also found that ammonia and nitrite con-
centrations were higher, and feed utilization and survival
rates were lower, in shrimp (L. vannamei) PL in BFT sys-
tems than in clear-water RAS. Alkalinity concentration was
also much higher in the biofloc tanks, indicating denitrifi-
cation process. The authors suggested that poor water qual-
ity in the BFT system may have contributed to the lower
shrimp production. They also confirmed that microbial
communities in BFT systems are more difficult to control
and manage than in RAS. These results suggest that bioflocs
may not provide nutritional contribution to farmed
shrimp. Meanwhile, when L. vannamei were nursed in clear
water (CW) and biofloc systems, the overall water quality
parameters (ammonia, nitrite, nitrate and pH) were better
in CW than in BFT system, presumably due to the elimina-
tion of solids in the CW treatment and using an external
biofilter (Ray et al. 2017).
The negative effects of bioflocs on productivity and water
quality have also been reported in farmed fish fry. For
example, when Nile tilapia were reared in chemoau-
totrophic BFT (no addition of carbon sources except feed)
and clear-water RAS, survival rates, growth performance,
feed utilization and water quality parameters were signifi-
cantly better in RAS systems than in the BFT system (Fleck-
enstein et al. 2018). Similar results were also reported in
mullet (Mugil cephalus) and Tinca tinca reared in hetero-
trophic BFT or RAS systems (Vinatea et al. 2018).
Decreased performance in BFT systems has been attributed
to poor water quality conditions.
Integrated biofloc-based shrimp culture
One of the major problems in the BFT system is the exces-
sive precipitation of solids, which are accumulated on the
bottom of culture units (Ray et al. 2010b). Solid precipita-
tion generally leads to water quality deterioration and
impairing the performance of cultured fish and shrimp,
mainly due to the increased levels of nitrogenous com-
pounds (Emerenciano et al. 2013a,b). Heavy solids may
also cause clogging of fish and shrimp gills (Schveitzer et al.
2013b). Therefore, the integration of different aquatic spe-
cies, occupying different trophic levels in the food chain,
has been proposed as an alternative way of minimizing
accumulated wastes, and improving water quality and pro-
ductivity and shrimp performance. In this regard, a num-
ber of studies have investigated the integration of shrimp
farming in BFT systems with other plant or animal species.
Poli et al. (2019a) demonstrate that integrating white
shrimp and Nile tilapia in a BFT system can increase the
694
total yield by up to 31.2%. This system can also decrease
sludge: biomass ratio and enhance nitrogen and phospho-
rus recovery and, overall, increase the sustainability of L.
vannamei culture in BFT systems. Similar results were also
found by Martins et al. (2020), who reared Pacific white
shrimp in integration with Nile tilapia in biofloc hetero-
trophic treatment (in which carbon was added throughout
the trial and no biofloc was inoculated at the beginning), or
in a mature treatment (where half of the volume of shrimp
rearing units was filled with biofloc-rich water). The
growth of shrimp was not significantly affected in all treat-
ments, while Nile tilapia reared in the heterotrophic treat-
ment showed better performance. The N and P recovery
was also higher in the heterotrophic treatment, while TAN
and nitrite were lower in the mature treatment. These
results suggest that the organic carbon supplementation
can enhance Nile tilapia growth and health in the inte-
grated system, and maintain good water quality.
A couple studies were also carried out on the integration
of shrimp and mullets (genus: Mugil) in BFT systems.
Mugil species are mud feeders, feeding mainly on bottom
detritus, microorganisms, such as algal aggregates, bacteria,
ciliates, rotifers and nematodes (Rao & Babu 2013; Mondal
et al. 2015). This means that these fish are an ideal candi-
date for feeding on bioflocs. In support, Borges et al.
(2020) grew mullet Mugil liza and white shrimp L. van-
namei either in monoculture systems or in integrated BFT
in separated tanks or in the same tank. The yield of shrimp
in monoculture system was higher than that in integrated
systems, meaning that the presence of mullets reduced
shrimp performance. However, the growth of shrimp
reared with mullet in the same tank or in separate tanks
was not significantly different. The sludge in the integrated
BFT systems was reduced, indicating that produced bioflocs
were efficiently consumed by mullet. Mullets also reduced
the number of Vibrio spp. in the system. The results of this
study simply mean that mullet and shrimp can be raised in
the same rearing unit, with no need for additional rearing
facilities.
The integration of white shrimp and another mullet spe-
cies (Mugil curema) in a BFT system was also evaluated
(Legarda et al. 2019). The shrimp were fed on 35% cp com-
mercial feed using preset feeding table, while mullet were
offered a 40% cp feed, once a day, at 1% of their biomass,
to stimulate biofloc consumption. The growth and survival
of shrimp reared separately or in integration with mullet
were not different, but the total yield significantly increased
in the integrated treatment. On the other hand, mullet
raised in the integrated system exhibited adequate growth
and survival rates and immune response. The shrimp-mul-
let integration led to a significant increase in phosphorus
recovery compared to shrimp monoculture. Thus, these
results suggest that the production of white shrimp and
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© 2020 John Wiley & Sons Australia, Ltd

mullet (Mugil curema) in integrated, BFT-based system is
viable, but long-term trials at commercial scale are needed
to verify the cost/effectiveness of this system.
Integrated multitrophic shrimp culture
Integrated multitrophic aquaculture (IMTA) is an aquacul-
ture system in which two or more aquatic species of different
trophic levels are farmed together in the same environmental
system, where the wastes of one species are recycled to
become inputs for another (Chopin et al. 2001; Granada
et al. 2016). Aquatic plants in IMTA can uptake substantial
amounts of phosphorous and nitrogenous compounds
resulting from farmed shrimps, feed wastes and dead plants.
Therefore, this integration can improve water quality and
performance of cultured species, reduce the environmental
impacts of aquaculture, provide economic benefits and,
overall, promote the ecosystem approach to sustainable
aquaculture (Granada et al. 2016; Rosa et al. 2020).
A number of studies evaluated shrimp farming in IMTA
systems, with aquatic plants and/or animals, with encour-
aging results. The growth and feed utilization of white
shrimp L. vannamei reared in a heterotrophic BFT system
in integration with green seaweed Ulva lactuca or with red
algae (Gracilaria birdiae) significantly increased (Brito et al.
2014; Brito et al. 2016). Shrimp/algae integration also
reduced TAN, NO2-N, NO3-N, phosphate and Vibrio den-
sity compared to seaweed-free biofloc. Brito et al. (2018)
found also that when L. vannamei were fed low (32% cp)
or high (40% cp) diets in an integrated biofloc system with
the seaweed G. birdiae, or in a monoculture system using
low protein, commercial feed (32% cp), growth rates were
similar in the three treatments. This means that integrating
white shrimp with G. birdiae in a BFT system can reduce
dietary protein levels by about 20% without adversely
affecting shrimp performance.
The efficiency of shrimp/algae integration may depend on
the density of farmed shrimp and/or algae. Appropriate den-
sities will likely lead to better nutrient recycling, better water
quality and higher performance. When L. vannamei (25 and
50 shrimp m 2) and G. corticata (0, 200 and 400 g m 2)
were reared in IMTA system under zero-water exchange, the
best shrimp performance and survival were obtained at 25
shrimp m 2 and 400 g seaweed m 2 (Fourooghifard et al.
2017). On the other hand, the lowest performance was
recorded at 50 shrimp m 2 in the absence of seaweed. The
growth of G. corticata was also significantly higher at 25/400
(shrimp/seaweed) than at other densities. Increasing shrimp
density resulted in an increase in total ammonia, nitrite,
nitrate and phosphate levels in culture water, while the con-
centrations of these compounds declined with increasing
algal density at all shrimp densities (Fourooghifard et al.
2018).
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Use of bioflocs in shrimp aquaculture
The performance and efficiency of an IMTA system,
including three different species occupying different
trophic levels, L. vannamei, Nile tilapia (Oreochromis niloti-
cus) and Sarcocornia ambigua, in a BFT-based system, were
also investigated (Poli et al. 2019b). Shrimps were fed
according to the feeding table, while tilapia were fed with
only 1% of their biomass per day, to stimulate them to con-
sume bioflocs as natural food sources. No significant differ-
ence was found in the performance of tilapia and shrimp in
all treatments, with the exception of IMTA where the total
yield was significantly higher than that in the control treat-
ment (absence of Sarcocornia). On the other hand, IMTA
system significantly reduced nitrates concentration. These
results demonstrate that the application of IMTA can
increase the yield of the three species by up to 21.5%.
Economic considerations
Despite the high potential of biofloc technology in aquacul-
ture, as mentioned in the previous sections, it has some
drawbacks. The most important economic problem is the
excessive use of energy for continuously high aeration and
water mixing. This system also requires monitoring, alarms
and emergency power supply. This means that biofloc sys-
tems can increase operating costs, due to the cost of aera-
tion system and the carbon source added to the system
(P
erez-Rostro et al. 2014). Therefore, it is necessary that
economic analyses be performed before adopting the BFT
on commercial scales. In this regards, a number of studies
were carried out on the cost effectiveness of BFT in shrimp
aquaculture.
Several factors affect the cost effectiveness and viability
of shrimp culture in BFT systems, including larval and
grow-out survival, shrimp price, stocking density, initial
investment, growth rates and feed efficiency and culture
conditions. For example, Hanson et al. (2009) reported a
saving of 9% in shrimp (L. vannamei) in a BFT-based sys-
tem, when the stocking density was increased by 20% com-
pared to a baseline scenario (50 PL m 3). About 14% profit
was also gained when grow-out survival was increased by
20%. Almost similar savings were achieved when Malaysian
prawn Macrobrachium rosenbergii were raised in a biofloc
system (P
erez-Rostro et al. 2014). In the meantime, Avnim-
elech (2015) suggested that farming L. vannamei in biofloc
systems can result in 30% reduction in the operating costs,
due to the decrease in the use of commercial feeds, and the
dependence on natural food produced in the biofloc sys-
tems. The integration of Nile tilapia (O. niloticus) and green
tiger shrimp (P. semisulcatus) in a BFT system resulted in
33% reduction in the production costs (Megahed 2010).
Pinto et al. (2020) found also that farming juvenile L. vam-
mamei in BFT systems was zootechnically and financially
viable.
695
A. M. El-Sayed
The economic viability of adding maize starch to IMTA
system comprising white shrimp (L. vannamei), spotted
scat (Scatophagus argus) and water spinach (Ipomoea aquat-
ica) in BFT system was analysed (Liu et al. 2014). The addi-
tion of starch significantly increased benefit-cost ratio
(BCR) compared with starch-free group. In addition,
IMTA resulted in better BCR than IMTA receiving 80% of
the proposed feeding level. Xu et al. (2018) evaluated the
effects of two commercial diets, a less expensive (used
mainly for semi-intensive L. vannamei culture) and a more
expensive diet (used for intensive culture) on the perfor-
mance and economic viability of juvenile L. vannamei
reared in BFT system at different C/N ratios. The feed and
variable costs analysis suggest that the less expensive feed
was more profitable than the more expensive diet, espe-
cially at a C/N ratio of 12:1.
Biofloc technology does not only affect the direct eco-
nomic profitability of farmed shrimp, but it can also affect
the quality of the final harvest. For instance, the muscle
firmness of white shrimp (L. vannamei) was greater in
BFT-grown shrimp than in clear-water system (Chan-Vivas
et al. 2019). This, in turn, improved the final product qual-
ity and acceptability in the market. Nonetheless, more
research is needed in this area to evaluate the effects of bio-
floc on other quality issues such as shelf life, rigour mortis
and physicochemical changes of refrigerated and frozen
shrimp.
Commercial application
The first commercial applications of BFT in shrimp aqua-
culture were reported in Belize in mid-1990s, with a pro-
duction of 11–26 mt of shrimp ha 1 per cycle, using 1.6 ha
lined outdoor ponds (McIntosh et al. 1999; Taw 2010;
Emerenciano et al. 2013a). Commercial biofloc ponds vary
in size, from 0.1 to 2 ha (Browdy et al. 2012). Aspirators
and paddle wheels are generally used to aerate and mix the
pond water to keep particles in suspension. Currently,
commercial, large-scale and small-scale BFT-based shrimp
farms are expanding in a number of countries, especially in
Asia (e.g. Indonesia, Malaysia, Thailand, South Korea,
China and India) and the Americas (e.g. USA and Brazil)
(Emerenciano et al. 2013a; Thong 2014). In Indonesia, for
example, about 20–25% of shrimp farms employ biofloc
technology, with an average production of more than 20
mt ha 1 per cycle in commercial, lined 0.5-ha ponds (Taw
2010). Ponds are partially or completely lined with high-
density polyethylene (HDPE) sheets. Aeration capacity is
28 hp ha 1, whereas paddle wheel aerators are placed in
two rings parallel to dykes and one at the centre of the
pond. Average stocking density is 130 PL m 2 or higher. In
Malaysia, lower production was achieved (12 mt ha 1 per
696
crop) when only pond dikes were lined, while fully lined
ponds produced 16.2–22.5 mt ha 1 (Taw et al. 2011).
Despite the wide expansion of commercial intensive and
super-intensive shrimp culture in biofloc systems, data and
information on the economy of these systems are almost
lacking, or not accessible to the public. Experience and
information generated by private BFT enterprises are gen-
erally not open to the public (Avnimelech 2015). These
companies tend to keep information and own knowhow
proprietary. Even when the data are available, proper dis-
semination channels are almost lacking.
Challenges and constraints
Researches on biofloc technology have been underway for
over three decades, while commercial applications have
been in place since the mid-1990s. But fundamental issues
related to BFT system function are still not well understood
(Taw 2010; Hargreaves 2013), and the BFT systems still face
several drawbacks and difficulties. One of the most serious
challenges facing the sustainability of BFT systems is the
continuous fluctuation in water quality. When outdoor sys-
tems are exposed to sunlight, microalgae bloom, leading to
fluctuations in pH, DO, CO2, ammonia and TSS (Harg-
reaves 2006). Consequently, the performance of these sys-
tems varies considerably overtime. This indicates the
sensitivity of BFT systems to culture conditions, such as
water quality (nutrient contents, pH, DO, CO2, ammonia,
temperature, etc.), light intensity, site specificity, inconsis-
tency of production, and diversity of the biomass and the
complexity of their population dynamics (Thong 2014).
Therefore, developing operational guidelines for these
divergent and complicated systems is difficult. As a result
of these drawbacks, many biofloc-based farms went out of
business or switched to other farming systems, presumably
due to the lack of technical experience on how to manage
these complex systems.
The high cost of farming inputs, especially the cost of
energy for aeration, water mixing and suspension of
organic particles, remains a major constraint limiting the
commercial expansion of BFT for shrimp aquaculture
(Avnimelech 2015). Also, power failure can lead to the col-
lapse of the system, causing dramatic economic losses. The
slow development of initial biofloc system is another issue,
since it may take more than 4 weeks for the establishment
of nitrifying bacteria within the BFT system. The off-fla-
vours of BFT-produced fish and shrimp (caused by turbid-
ity and cyanobacteria) may pose a serious concern for their
marketing (Avnimelech 2015). Purging this off-flavour by
keeping the fish and shrimp in a clean, running water for a
certain time prior to harvest, may be necessary, despite the
extra cost of this process.
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Eventually, full understanding of the BFT systems is
essential for better management, sustainability and cost
effectiveness of these systems. Farmers and practitioners
should be trained on how to adopt the successful experi-
ences of BFT technology gained over the years, with
emphasis on the economic and environmental benefits of
this technology (Crab et al. 2012). Continuous and profes-
sional monitoring of the BFT system performance is also
necessary for successful implementation of this technology
in shrimp aquaculture. More research and development
programs should also be dedicated to addressing the tech-
nical problems of BFT systems, in order to reduce their
operational costs and improve production capacities.
Cooperation between the private sectors and official insti-
tutions, organizations and NGOs should also be encour-
aged in BFT transfer, with emphasis on the successful
stories.
Conclusion
Biofloc is a symbiotic process, including farmed aquatic
animals, heterotrophic bacteria and other microbial species
in the water. Through this process, ammonia is removed
from culture system and the waste materials may be cycled
into microbial flocs. Nitrogen conversion process includes
three integral pathways for ammonia removal; they are as
follows: photoautotrophic removal by aquatic plants, auto-
trophic bacterial conversion of ammonia-nitrogen to
nitrate-nitrogen, and heterotrophic bacterial conversion of
ammonia-nitrogen directly to microbial biomass. In the
third pathway, the addition of suitable amounts of organic
carbon source (such as molasses, simple sugars, brans) is
required for stimulating heterotrophic bacterial growth. At
appropriate carbon-to-nitrogen (C/N) ratios, heterotrophic
bacteria assimilate ammonia-nitrogen into cellular protein.
These bioflocs can be used as natural food sources for cul-
tured organisms, in addition to improving animals’ health
status and water quality. A number of biofloc technology
(BFT) systems have been developed, including suspended-
growth systems, attached-growth biofiltration system, mov-
ing bed biofilm reactor and periphyton technology. The use
of any of these systems depends on farm site, farming
intensity and technical protocols.
Several factors affect shrimp production in BFT systems,
including shrimp species and stocking size and density,
water temperature, salinity, alkalinity, hardness, pH, aera-
tion, light intensity and photoperiod, presence of sub-
strates, carbon sources applied, carbon/nitrogen (C/N)
ratio and total suspended solids. Shrimp survival and
growth performance generally decrease with increasing
stocking density. Increasing stocking density will result in
excessive nutrient inputs, which need to be recycled by
microbial communities. These nutrients can cause
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Use of bioflocs in shrimp aquaculture
eutrophication of the systems and develop microbial bio-
mass. At high stocking density, vigorous aeration is
required to maintain a high particle density and to keep the
organic matter in suspension. A C/N ratio of 10-20: 1 has
been suggested for optimizing bacterial growth and pro-
duction of microbial flocs and, in turn, increasing growth
and production of BFT-reared shrimp. The presence of
light in floc production systems (especially in outdoor sys-
tems) is also essential for natural food production for
farmed fish and shrimp. Light is the limiting factor for
enhancing photosynthesis and phytoplankton blooming in
eutrophic outdoor ponds. Photosynthesis leads to diel fluc-
tuations in the concentrations of DO, CO2, pH and un-ion-
ized ammonia, rendering biofloc systems under continuous
variations between day and night. The addition of sub-
mersed substrates in BFT-based shrimp culture may also
improve water quality, reduce ammonia and nitrite con-
centrations, increase natural productivity, enhance periphy-
ton formation and provide natural foods for farmed
shrimp and improve their growth rates and immune
responses. Total suspended solids (TSS) are also an impor-
tant factor affecting biofloc production in intensive and
super-intensive BFT aquaculture. Excessive supplemental
organic carbon, with high C/N ratio, can cause rapid
increase in TSS. Under this situation, autotrophic biofloc is
shifted to heterotrophic microbial production, leading to
significant changes in water quality and biofloc density and
composition.
Bioflocs have good nutrient profiles, making them a
potentially good natural food source for farmed shrimp.
However, the nutritional value of biofloc varies consider-
ably, depending on the C/N ratio of culture water, dietary
feed composition and level, light intensity, composition
and age of biofloc aggregates, TSS, bacteria-phytoplankton
concentration and the culture conditions. Farmed marine
shrimps and freshwater prawns can readily consume bio-
flocs as a natural food source, or partial replacer for shrimp
diets or dietary protein sources. Biofloc consumption can
lead to significant improvements in shrimp performance,
survival rate, digestive enzymes activity, immune system,
health status and water quality, in addition to a significant
reduction in feed costs. Therefore, BFT has been widely
tested in shrimp culture during larval, nursery and grow-
out phases, and can also be applied in integration with
other aquatic animals and/or plants.
Despite the wide expansion and economic cost effective-
ness of commercial intensive and super-intensive shrimp
culture in biofloc systems, reliable information on the
economy of these systems is almost lacking or not accessi-
ble to the public. In addition, the BFT systems still face sev-
eral drawbacks and difficulties. Continuous fluctuation in
water quality parameters, high cost of farming inputs,
power and mechanical failure, slow development of initial
697
A. M. El-Sayed
bioflocs and off-flavours of BFT-produced fish and shrimp
are the major challenges facing the sustainability of biofloc
technology in shrimp aquaculture. Therefore, full under-
standing of the BFT systems is essential for better manage-
ment and, in turn, for sustainability and cost effectiveness
of these systems.
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