Aquaculture International (2024) 32:9321–9374

https://doi.org/10.1007/s10499-024-01618-w

REVIEW

Water quality in biofloc technology (BFT): an applied review

for an evolving aquaculture

Mohammad Hossein Khanjani1 · Alireza Mohammadi2 ·

Maurício Gustavo Coelho Emerenciano3

Received: 2 June 2024 / Accepted: 7 July 2024 / Published online: 19 July 2024
© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2024

Abstract
Biofloc technology (BFT) has been identified as an effective and sustainable aquaculture
method, particularly beneficial for warm-water species in tropical areas. This technology
is widely used in intensive aquaculture for several aquatic species due to its capacity to
significantly reduce water exchange rates, benefiting both production systems and the envi-
ronment. The efficacy of BFT in farming operations is directly related to a proper manage-
ment of water quality parameters within the optimal range of the target species, as these
parameters directly impact the yield of production units. Essentially, BFT functions as a
water quality management system, converting harmful nitrogenous waste such as ammonia
and nitrite into less harmful forms such as nitrates through microbial activity, ensuring
the health of aquatic organisms. Key environmental factors such as temperature, dissolved
oxygen (DO), pH, salinity, alkalinity, TAN, nitrite, nitrate, settleable solids (SS), and total
suspended solids (TSS) can affect the growth of aquatic species and the functionality of the
microbial community. This review brings (i) a comprehensive bibliometric analysis on bio-
floc and water quality, (ii) highlights optimal ranges, and (iii) key observations of several
water parameters including temperature, salinity, nitrogenous compounds, SS, TSS, DO,
phosphate, pH, and alkalinity in BFT rearing conditions for key aquatic species. Elements
such as countries, thematic and keywords, and authors were explored, correlated, and dis-
cussed. In addition, this manuscript also (iv) discusses the presence of heavy metals and
microplastics (MPs) in BFT culture water. A dedicated review on water quality in biofloc
technology will contribute to future research and development (R&D) in this topic, support
decision-making to improve farming operations, and can help further expansion of BFT-
based aquaculture.

Keywords Aquaculture · Biofloc · Total suspended solids · Nitrogenous compounds ·

Heavy metals · Microplastic · Bibliometric analysis

Handling Editor: Brian Austin

Extended author information available on the last page of the article

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Introduction

The increasing aquaculture sector is expected to be a key contributor in addressing the

growing demand for protein by the growing global population (FAO 2024). This sector
incorporates a variety of culture systems, suitable for both small and large operations.
These systems range from net pens in marine environments and deep lakes to earthen
ponds, raceways, tanks, and a variety of recirculation setups. Each type of system is
associated with its own unique ecological challenges. These challenges may include the
discharge of surplus organic materials into nearby ecosystems, the emission of antibiotic
and antiparasitic compounds into the environment, the spread of pathogens into farmed
and wild populations, and concerns regarding the welfare of the animals (Carballeira
Braña et al. 2021; Strand et al. 2022; Bagi et al. 2023). In regions where traditional
land-based aquaculture is prevalent, and where the implementation of a clear-water
recirculating aquaculture system (RAS) is prohibitively expensive, other land-based
limited water exchange systems have been developed and adopted (Zhang et al. 2011).
Among these, biofloc technology (BFT) has gained prominence in recent decades as an
effective and economical method for cultivating a broad range of species in warmer and
temperate regions (Hargreaves 2013; Yu et al. 2023a, b; Khanjani et al. 2024a). Initially,
BFT was designed to minimize the environmental footprint of land-based aquaculture,
primarily serving as a system for waste management and disease prevention (Khanjani
et al. 2024b). BFT effectively maintains water quality for species, thereby lowering the
demand for adding new water to aquaculture systems (Ekasari et al. 2015; Samocha
et al. 2017). However, the application of BFT has been limited, as not all species are
suitable for culture in BFT systems (Emerenciano et al. 2013). Certain biological traits
are essential for organisms to adapt and overcome stress factors such as moderate levels
of dissolved oxygen (DO), especially at high biomass (Emerenciano et al. 2013) and
high stocking densities (Lima et al. 2019). Ideally, the selected species must be able to
tolerate fluctuations in nitrogenous compounds (Widanarni et al. 2012), settleable solids
(SS), and total suspended solids (TSS) (Browdy et al. 2012). Once all these aspects are
taken into consideration, the minimal or zero water exchange in BFT systems contrib-
utes to water quality management, which fosters the growth and proliferation of key bac-
teria and other microbial communities (Martins et al. 2019). Changes on animal behav-
ior and performance typically indicate the deterioration of water quality and recall the
necessity for management intervention (Bagi et al. 2023). For instance, these behaviors
may suggest increased ammonia or nitrite levels or reduced dissolved oxygen, prompt-
ing the need to enhance aeration, adjust key parameters such as alkalinity (Furtado et al.
2015), and modify microbial management. Thus, monitoring environmental parameters
is crucial in BFT systems (Alkhamis et al. 2023). A comprehensive understanding of
water quality parameters and their interconnection is vital for the success and mainte-
nance of BFT-based aquaculture operations. For aquatic species to thrive, it is important
to maintain environmental parameters such as DO, pH, TSS, total ammonia nitrogen
(TAN), nitrite, and alkalinity within an optimal range to ensure proper performance and
survival (Alkhamis et al. 2023). Numerous studies have been conducted to investigate
how BFT impacts water quality (Pellegrin et al. 2022; Schveitzer et al. 2024). BFT has
been shown to be more effective in removing inorganic nitrogen from water compared
to traditional methods (Emerenciano et al. 2017). BFT also enhanced the growth of
species such as catla (Catla catla), rohu (Labeo rohita), and mrigal (Cirrihinus mri-
gala) in polyculture systems (Deb et al. 2020). Additionally, the successful culture of

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Penaeus vannamei and Farfantepenaeus brasiliensis postlarvae in BFT is explained

by water quality and the use of the system’s microbes as a food source, contributing
to tissue development, health, and improved nutrition (Emerenciano et al. 2012; Suita
et al. 2015). Routine monitoring, interpretation, and intervention of key water quality
parameters are essential for maintaining a suitable environment, animal welfare, health,
and performance, especially in BFT with high stocking density. However, in commer-
cial biofloc-based shrimp and fish operations, farmers still struggle with water quality
management, especially due to lack of understanding and proper follow-up actions (Day
et al. 2016; Emerenciano et al. 2022a). In this review, a comprehensive bibliometric
analysis on biofloc and water quality is presented. The manuscript highlights optimal
ranges and key observations of several water quality parameters including temperature,
salinity, nitrogenous compounds, SS, TSS, DO, phosphate, pH, and alkalinity in BFT
rearing conditions for key aquatic species; and discusses the presence of heavy metals
and microplastics (MPs) in BFT culture water. Finally, this review aims to (i) contribute
to a more “holistic” understanding of BFT, with a key focus on water quality parameters
and relevant management implications; and (ii) support decision-making to improve
farming operations, and (iii) boost further R&D development of BFT-based aquaculture.

Bibliometric

Material and methods

This study aimed to review the research conduct on water quality in BFT. The Scopus
database was accessed for works published between 1989 and 2024 using the following
keywords (“biofloc” OR “biofloc system” OR “biofloc technology” AND “water qual-
ity” OR “dissolved oxygen”, OR “temperature”, OR “salinity”, OR “heavy metal”). In
total, 802 publications were found in the literature. Complete records and their refer-
ences were extracted to be used as bibliometric data. The research process involved five
steps including (i) gathering data, (ii) selecting the appropriate software, (iii) analyzing
the data, (iv) visualizing the results, and (v) interpreting the results.
The data was analyzed and visualized using Excel 2019, VOSviewer version 1.6.16,
and the Bibliometrix package for R (Aria and Cuccurullo 2017). For each of the records,
author, country, and year of publication were directly accessed from the Scopus data-
base (www.​scopus.​com/​sourc​es), and maps were created in Excel. VOSviewer was uti-
lized to create the network of countries, journals, and keywords, including author and
index keywords (Gao et al. 2018). Finally, the quality of the records was checked in
Microsoft Excel, and duplicates, spelling errors, and incomplete records were corrected.
Items that failed quality control were excluded from the study.

Three‑field plot

Relationship between three elements including countries, keywords, and authors using
Sankey plots was plotted. Three-field plots utilize colored blocks to represent the key
findings of the analyses. The total number of links between the blocks determines their
height.

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Thematic map and evolution

For the thematic map, four clusters were identified and developed: (i) low density and cen-
trality (emerging themes): The topics placed in the bottom left quadrant need further explo-
ration due to underdevelopment or being emerging subjects. These subjects enjoy limited
significance across the network and their development is slow within the network; however,
they present attractive opportunities for future research. (ii) High density and low central-
ity (basic themes): The topics located in the top right quadrant constitute the primary axes
in the body of research and are central to the developments in the field. These topics enjoy
high relevance across and inside networks. By identifying and developing these topics,
bibliometric studies highlight the opportunities for further investigation. (iii) High density
and low centrality (niche themes): Topics in the top left quadrant are internally well-con-
nected but have limited external connections, indicating their limited significance under
current circumstances. (iv) High density and centrality (motor themes): Topics in the top
right quadrant represent the major trends in the field and are central to the literature. They
have high relevance and connectivity within and across the network. Given their centrality,
bibliometric studies can shed light on their exact nature and guide future research. In the
thematic map, the bubble sizes are determined by the number of publications in which the
keyword appeared (García-Lillo et al. 2023).
For the R&D evolution, Sankey diagrams were utilized to illustrate themes and their
development over time regarding water quality in biofloc technology (Xiao et al. 2022).
In the Sankey plot, each box corresponds to a theme, with the size of the boxes indicating
frequency (Xiao et al. 2022). The lines connecting the boxes show the evolution of a theme
with line thickness corresponding to link strength.

Results

The number of “biofloc” publications increased exponentially between 1989 and 2024.
The “biofloc and water quality” ones first exceeded 50 publications in 2017 (Fig. 1).
These publications grew on average by close to 6.84% from 1989 to 2024 (Fig. 1). The

Fig. 1  Number of publications per year, as per bibliometric analysis of “biofloc and water quality” (1989–
2024)

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least active years were 1990,1991, 1993–2003, and 2005–2006 while the peak occurred
in 2021 with 127 papers. As shown in Fig. 1, 802 studies were extracted from Scopus
across 190 sources between 1989 and 2024. Overall, 2460 authors had contributed to
the literature on the topic. The majority of the documents were general research articles
(n = 710), followed by reviews (n = 36). The vast majority of the records were in English
(95.7%).
Figure 2 provides a detailed graphical overview of research collaboration across the
world. The top countries in terms of productivity of the field are Brazil, China, and
India, respectively. The highest frequency of international collaboration was between
Brazil and Mexico (n = 198), followed by Brazil and USA (130), China and Malaysia
(90), Brazil and Australia (80), and China and USA (75). The blue colors in Fig. 2 indi-
cate different scientific publications in the field of biofloc, whereas darker ones indicate
higher number of publications.

The three‑field plot

The flow in the three-field plots indicates that authors from Brazil, China, and Mexico
have published the largest number of works on the subject (Fig. 3). As shown in the
figures, there are geographical preferences for certain keywords. For instance, there is a
strong relative preference among authors from Brazil for “biofloc”, “shrimp”, “P. van-
namei”, and “Oreochromis niloticus”; and for authors from China, “biofloc” and “bio-
floc technology” were preferred.

Fig. 2  Countries’ scientific production (A) and countries’ collaboration map (B) in the field

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Fig. 3  Three-field plot for county of the corresponding country, keywords, and authors, as per bibliometric
analysis of “biofloc and water quality” (1989–2024). The height of the rectangles corresponds to the sum of
the relations originating between the elements and the thickness of the links represents the strength of the
connections

Thematic map and evolution

According to the results of thematic map, biofloc, shrimp growth, probiotics, nursery, and
heterotrophic bacteria are the motor themes. Moreover, aquaculture, sustainability, nitrifi-
cation, biofloc system, microalgae, and O. niloticus are emerging themes that require fur-
ther development (Fig. 4).
From a general perspective, it can be found that “biofloc” is present in all times. It is
noticeable that the terms “water quality” and “nitrification” were first used in 2020–2024.
In the following 4 years, they were further developed and sustained interest in the final
period of 2022–2024. (Fig. 5).

Biofloc technology (BFT)

Research and practical application of BFT have been ongoing since the 1970s, start-
ing at Ifremer-COP and reaching commercial viability by the late 1980s (Emerenciano
et al. 2013). BFT as an aquaculture production system is evolving quickly; however, its
core still emphasizes the growth of a large microbial population in water, leading to the
formation of microbial aggregates known as “bioflocs” (Khanjani and Sharifinia 2024).
These living microbial particles play a crucial role in controlling nitrogenous com-
pounds in the culture water (Emerenciano et al. 2017). Both BFT and RAS utilize bio-
logical processes to achieve nitrogen removal, but their approaches differ significantly
(Liu et al. 2021). For instance, besides the chemoautotrophic communities (nitrifying
bacteria), BFT also relies on heterotrophic microorganisms for toxic N-compounds con-
trol (Khanjani et al. 2022). In addition, the typical “clear-water” environment found in
RAS is not the case in BFT where some levels of suspended solids are expected but
must be maintained within acceptable levels for the different aquatic species (Gaona

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Fig. 4  Thematic map in “biofloc and water quality” research

Fig. 5  Thematic evolution in “biofloc and water quality” research

et al. 2017; Emerenciano et al. 2017). Also, BFT systems are normally less expensive to
set up and run than RAS, as well as offering several other biological benefits over tradi-
tional clear-water RAS (Liu et al. 2019).
There has been a marked increase in publications on BFT in aquaculture since 2015.
These publications are not only related to laboratory scale, but also carried out in large
commercial operations (Emerenciano et al. 2022a). In this sense, BFT is increasingly
viewed as a sustainable alternative in aquaculture, particularly in tropical shrimp farm-
ing (Ulloa Walker et al. 2020; Emerenciano et al. 2021; El-Sayed 2021). One of the
key attributes is the in situ circularity, in which bioflocs are able to control the nutrient
content in the water by converting waste nitrogen into microbial biomass and serve as

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supplemental feed for the cultured species (Khanjani et al. 2023a). Moreover, bacteria
in the bioflocs offer additional health benefits by outcompeting pathogenic microbes
and acting as natural probiotics (Deng et al. 2022; Bagi et al. 2023). Therefore, BFT is
seen as a new chapter in the “blue revolution” (Stokstad 2010). Facilitated by minimal
water exchange and circularity properties, BFT continuously boosts the recycle and
reuse of nutrients within the culture medium (Raza et al. 2024). The system’s sustain-
ability is further highlighted by (i) its high production efficiency and small footprint
(da Silveira et al. 2020); and (ii) nutrition attributes due to improved feed efficiency as
a result of continuous ingestion of protein and lipid-rich microbial aggregates (Khan-
jani et al. 2024b). This natural productivity (bioflocs) is crucial in preserving the water
quality. Microorganisms are the central pivot in BFT operations (Emerenciano et al.
2021). Water quality is controlled through encouraging mainly heterotrophic and nitri-
fying bacterial growth and regulating photoautotrophic communities by maintaining
among others (i) a suitable carbon-to-nitrogen (C:N) ratio. This ratio is essential for
optimal growth of heterotrophic bacteria, which utilize carbon for maintenance of
essential activities such as respiration, feeding, digestion, and reproduction (Emeren-
ciano et al. 2021; Khanjani et al. 2023b, 2023c). But also (ii) high DO and alkalinity
levels to ensure a proper nitrification process (Furtado et al. 2015).
In terms of C:N ratio management in BFT, it typically involves two phases (Emeren-
ciano et al. 2021). Firstly, the development phase employing a specific C:N ratio (normally
10–20:1) and addition of an external carbon source such as grains and sugars (Wei et al.
2016); and secondly, the maintenance phase reducing or eliminating the external carbon
inputs, depending mainly on TAN, ­NO2 values, and desirable SS levels (Khanjani et al.
2024c).

Water quality parameters

A proper water quality management is crucial to the success of production cycles in aqua-
culture farms. BFT systems are particularly sensitive to fluctuations in factors such as tem-
perature, DO, pH, salinity, suspended solids, alkalinity, orthophosphate, and nitrogenous
compounds, but also other external factors such as light intensity and minerals. A holistic
view of these parameters and their interconnection is crucial for effective and sustainable
management. For that, Tables 1, 2, and 3 bring key information regarding BFT and water
quality management. Table 1 summarizes water quality parameters in different studies con-
ducted using a variety of aquaculture systems and BFT. Table 2 shows the suitable ranges
of water quality parameters in the biofloc aquaculture system. And finally Table 3 shows
the effects of manipulating different water parameters in BFT and the impact on animal
performance in several species.
It is important to note that each species demands specific conditions that require regu-
lar monitoring to promote growth and ensure survival. Thus, water quality has a profound
effect on the health of the organisms, survival rates, and the profitability of aquaculture
(Tumwesigye et al. 2022). While parameters such as salinity is often relatively constant in
the system, others such as alkalinity and DO may change, especially in high biomass, and
interventions are needed. An in-depth understanding of key water quality parameters and
their interactions in BFT is essential for decision-making and suitable interventions within
the production cycles.

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 Table 1  Comparative analysis of water quality in clear water and biofloc systems across different studies
Rearing Cultivated spe- T (°C) S (g/L) pH A (mg DO TSS TAN NO2 NO3 PO4-P Reference
system cies/life stage ­CaCO3/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L)

BFT P. vannamei/adult 30 34.2 7.1 126 4.0 546.7 0.38 0.23 22.9 54.5 Vinatea
et al. 2009
BFT P. vannamei/PL 29.16– 7.33–7.76 215.33– 6.61–6.94 190.92– 0.12–0.35 0.026– 6.97– 0.54–0.66 Lim et al.
31.28 273.33 304.38 0.035 10.73 2021
BFT P. vannamei/J 27.05– 33.5 7.5–7.90 100–162.2 4.9–5.1 850–890 0.3–0.4 3.1–4.0 57.5– 5–6.68 Furtado
27.25 60.53 et al. 2011
CW 27 33 6.86 78 5.1 870 0.6 4.3 54.5 8.2
BFT O. niloticus/J 27.55– 6.56–8.27 44.72– 5.69–6.04 485– 0.94–1.68 7.03–10.1 105–140 6.93– Martins
27.70 175.90 548.61 10.23 et al. 2019
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BFT Macrobrachium 26.34– 7.76–7.80 137.69– 6.48–6.98 298.7– 0.21–0.54 0.64–0.92 2.1–2.42 Islam et al.
rosenbergii 26.47 148.95 445.2 2023
CW 26.39 7.85 139.78 6.75 168.4 0.71 1.98 3.12
BFT Lates calcarifer/J 27.2–27.5 0 7.8–8 94.6–96.0 7.5–7.7 0.42–0.45 0.9–0.94 0.13–0.17 0.081– Whangchai
0.098 et al. 2022
BFT Clarias 7.3–7.76 177.13– 7.51–7.79 Chen et al.
gariepinus/J 263.53 2020
BFT Litopenaeus 27.49 5 8.50 7.61 7.25 0.11 1.37 87.34 Valenzuela-
setiferus/N 28.02 35 8.03 5.33 6.6 0.29 1.47 92.34 Jiménez
et al. 2022
BFT P. vannamei/J 28.38– 35.3–36 7.36–7.76 133.4– 4.49–4.91 280.16– 1.0–5.91 1.97–8.97 3.97– 2.7–3.67 Lara et al.
30.79 229.7 460.5 14.88 2017
BFT Colossoma 25.78 6.32 11.13 0.34 0.67 2.60 dos Santos
CW macropomum/J 25.97 6.97 6.67 1.98 0.59 5.09 et al. 2020
BFT P. vannamei/N 29.2 34.5–34.9 117–120 5.6 0.25–0.27 3.81–4.14 43.58– 4.83–5.03 Gaona et al.
44.63 2016
CW 29.06 35.43 113.98 5.47 0.31 4.18 54.72 5.34
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 Table 1  (continued)
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Rearing Cultivated spe- T (°C) S (g/L) pH A (mg DO TSS TAN NO2 NO3 PO4-P Reference
system cies/life stage ­CaCO3/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L)

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BFT Rhamdia 24.7–25.4 1.3–1.4 7.7–8.0 107.9– 6.5–6.8 538.4– 0.05–0.26 0.01–0.8 58.4–86.0 12.4–13.9 Poli et al.
quelen/F 187.5 157.0 2015
CW 25.35 0.3 7.7 83.7 6.7 - 0.11 0.02 20.3 6.9
BFT P. vannamei 29.7 33.0–33.7 7.5–7.8 133.3– 4.8–5.5 4.0–9.1 Schveitzer
218.4 et al. 2013
BFT P. vannamei 30.8 30.6 8.27 171.6– 7.49 304–618 0.27–0.29 0.23–0.32 7.21–8.83 Liu et al.
nauplius 174.1 2022
CW 30.7 31 8.29 167.7 7.5 213 0.5 0.03 3.30
BFT Piaractus 25.95– 8.00 96.21– 6.76–6.90 278.50– 0.25 3.33 48.13– Angeles-
brachypomus/F 26.10 103.27 363.3 59.63 Escobar
CW 26.01 7.74 58.37 6.82 7.20 0.33 0.31 21.73 et al. 2022
BFT P. vannamei/N 23.08– 25 76–7.8 138.68– 5.99–7.39 285.08– 0.24–0.57 0.61–0.93 2.09–2.56 Rajkumar
23.16 153.52 493.5 et al. 2016
CW 23.29 25 7.8 138.89 6.57 158.5 0.78 1.89 3.21
BFT P. vannamei/J 27.86– 37.23– 7.74–7.93 116.71– 5.68–5.85 306.37– 2.54–3.42 0.73– 0.38–1.73 0.51–4.82 Gaona et al.
28.17 38.49 159.29 745.2 10.28 2017
BFT O. niloticus/F 28.20 10.5 7.9 6.7–6.9 191.8 0.3 0.6 25.4 Fleckenstein
CW 28.35 10.3 7.8 6.9–7.0 132.1 0.2 0.1 22.3 et al. 2018
BFT P. vannamei/G 30.18 23.59 7.16 5.57 244.0 0.16 2.63 0.73 3.44 Pimentel
and N et al. 2020
BFT O. niloticus/J 27.54– 7.10–7.50 56.11–70 6.40–6.51 635–841.5 3.56–3.70 0.56–0.99 321.18– Eid et al.
27.68 466.82 2021
CW 27.54 8.03 48.89 6.58 588.64 2.20 0.80 450.30
BFT O. niloticus/F 27.55– 7.33–7.53 48.95– 5.91–5.97 501.47– 0.73–1.17 5.94–8.14 117.50– 1.75–8.30 Martins
27.70 75.76 707.49 143.33 et al. 2017
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 Table 1  (continued)
Rearing Cultivated spe- T (°C) S (g/L) pH A (mg DO TSS TAN NO2 NO3 PO4-P Reference
system cies/life stage ­CaCO3/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L) (mg/L)

BFT Labeo rohita/F 27.06 7.07 5.4 284.48 0.11 0.03 0.03 Kamilya
et al. 2017
CW 27.06 6.59 6.53 23.85 0.06 0.03 0.06
BFT Etroplus 25–25.3 0.095 7.66–7.69 7.43–7.65 1307– 0.008 0.23 0.2 0.36–0.39 Thilakan,
suratensis/J 1318 et al. 2019
CW 25.6 0.094 8.32 7.17 1249.57 0.01 0.20 0.22 0.37
BFT P. vannamei/PL 28.8–29.2 10.3–30.2 7.7–7.9 310–322 7.7–8.9 263–330 0.7–1.2 0.3–0.6 0.3–1.4 2.0–2.6 Ray and
Lotz 2017
BFT O. niloticus/J 27.57– 0–16 7.4–7.6 93–152 4.70–5.15 384.5– 3.94–5.84 0.82–8.89 40.58– 22.12– de Souza
27.77 566.9 65.58 35.01 et al. 2019
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BFT O. niloticus/F 23.2 0.3‰ 8.1 198.0– 7.1 154.9 1.6–2.0 0.5–1.1 6.8–13.6 5.7–7.6 Silva et al.
301.8 –435.6 2017
BFT O. niloticus/J 23.97– 7.72–7.85 159.7– 6.17–6.32 465.9– 0.14–0.25 0.10–0.29 44.76–142 Dilmi et al.
24.19 168.77 750.5 2022
CW 24.15 8.04 177.5 6.60 - 0.11 0.12 7.19
BFT Gibelion catla/F 26.74– 7.4–7.7 37.0–42.67 5.7–5.9 93.55– 0.22–0.26 0.019– 11.27– Solanki
26.82 140.68 0.035 15.81 et al. 2023
BFT P. vannamei/PL 28.45– 30.1– 7.87–8.15 124.10– 4.67–5.95 123.53– 0.46–0.86 0.14–0.39 0.10–0.22 0.52–0.98 Panigrahi
28.90 30.77 140.25 328.55 et al. 2019
CW 28.45 30.15 8.58 150.25 6.94 60.58 1.45 0.75 0.67 0.43
BFT P. vannamei/J 28.7–28.8 5.0‰ 6.9–7.7 104.0– 5.28–5.57 327.4– 1.3–7.1 1.9–14.0 4.6–77.0 Huang et al.
173.0 682.1 2022b

Abbreviations: T temperature, S salinity, A alkalinity, DO dissolved oxygen, TSS total suspended solids, TAN total ammonia nitrogen, PL postlarvae, J juveniles, N nursery, F
fry, G grow-out
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 Table 2  The suitable ranges of water quality parameters in the biofloc system
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Parameter Appropriate range Considerations Reference

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T (°C) 26–32 °C Optimal temperatures are species-specific. For instance, Ahammed et al. 2022; Emerenciano et al. 2017; Kim et al.
ranges from 26 to 28 °C are suitable for P. vannamei and 2019
tilapia species. Temperatures outside these ranges can
lead to reduced growth and disturbances in physiologi-
cal and behavioral activities. Excessive temperatures
can cause a decrease in oxygen levels and increase stress
during breathing
S (g/L) 4–33 The salinity levels are species-specific and must be kept Khanjani and Alizadeh, 2023; Khanjani et al. 2020a
according to the tolerance range. In biofloc systems, the
salinity level increases due to the limitation of water
replacement and the accumulation of uneaten food waste
pH 6.5–8.5 Levels outside these ranges can impact growth and lead to Emerenciano et al. 2017; Ahammed et al. 2022
disturbances in physiological and behavioral activities. In
biofloc systems, high TSS/SS and respiration rates may
cause pH reduction
Alkalinity (mg ­CaCO3/L) Higher than 100 mg/L High alkalinity minimizes the daily fluctuation in pH. In Van Wyk and Scarpa 1999; Furtado et al. 2015; Emeren-
addition, higher alkalinity levels (> 100 mg/L) are critical ciano et al. 2017
to support TAN processing by heterotrophic and chemo-
autotrophic bacteria
DO (mg/L) Higher than 5 mg/L Higher levels (> 5 mg/L) will safely sustain fish-shrimp Vinatea et al. 2009; Kuhn and Lawrence 2012;
physiological responses, as well as microbial develop-
ment. The stability of DO levels is dependent on several
factors including TSS, SS, salinity, and temperature
TSS (mg/L) 200–500 Species and age dependent (e.g., normally young individu- Schveitzer et al. 2013; Hargreaves 2013; Gaona et al. 2017
als are more resistant compared to their older counter-
parts). The level of TSS can significantly impact the
dissolved oxygen level, pH, and alkalinity. High levels
of TSS (e.g., > 500 mg/L) can impair the growth and
survival of several cultured aquatic species
SS (mL/L) 5–15 mL/L for Same as per TSS. High levels of SS (e.g., > 15 mL/L) can Emerenciano et al. 2017
shrimp, and 5–20 impair the growth and survival of several cultured aquatic
for tilapia species
Aquaculture International (2024) 32:9321–9374
 Table 2  (continued)
Parameter Appropriate range Considerations Reference

TAN (mg/L) Lower than 1 mg/L Dependent on factors such as species-age, temperature, pH, Ebeling et al. 2006a; 2006b;
and salinity. Three processes (photosynthesis, assimila- Emerenciano et al. 2017
tion, and nitrification) play a significant role in control- Khanjani et al. 2022
ling TAN in BFT
NO2 (mg/L) Lower than 1 mg/L Mainly dependent on factors such as species-age and salin- Vinatea et al. 2010; Emerenciano et al. 2017
ity. Nitrite-oxidizing bacteria are slow growers and quite
sensible to environmental changes
NO3 (mg/L) Lower than 200 mg/L Mainly dependent on factors such as species-age and Emerenciano et al. 2017; Gaona et al. 2017
salinity. For some species only toxic at higher levels
(e.g., > 400 mg/L). High concentration of TSS and SS
can boost nitrate levels in water
Aquaculture International (2024) 32:9321–9374

PO4 (mg/L) 0.5–20 mg/L Accumulation is expected BFT systems. Bacteria and Silva et al. 2013; Emerenciano et al. 2017; Liu et al. 2018
algae, as well as TSS and SS can play a role with ­PO4
fluctuations
Heavy metals (HMs) Unknown Salinity and feed inputs can play a key role on HMs accu- Deswati et al. 2022
mulation and toxicity

Abbreviations: T temperature, S salinity, A alkalinity, DO dissolved oxygen, TSS total suspended solids, TAN total ammonia nitrogen
9333

13
 Table 3  Manipulation of water quality parameters and impact on performance in biofloc technology in target aquaculture species
9334

Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
P. vannamei 1.91 WT (15, 21, 27, and 33 °C) P. vannamei shrimps were Shrimp reared at 15 and de Souza et al. 2016
stocked in the tanks at a 33 °C displayed higher
density equivalent to 200 activity of antioxidant
2
shrimps/m (or 40 shrimps enzymes as a defense
per tank), for 60 days mechanism against the
overproduction of ROS
triggered by temperature
stress
P. vannamei 6.5 WT (15, 21, 27, and 33 °C) The experimental setup Shrimp subjected to 15 and de Souza et al. 2014
comprised of 12 tanks, 21 °C temperatures exhib-
each with a capacity of ited increased enzyme
250 L. Shrimps were then activities. Moreover,
stocked in these tanks at a this adverse temperature
density of 150 shrimps/m2 condition might directly or
indirectly induce stress in
the animals
Paralichthys olivaceus 27.96 WT (20, 22, 24, 26, 28, and The experimental system The research shows that Kim et al. 2019
30 °C) included 12 tanks, each temperatures exceeding
with a capacity of 250 26 °C led to hematological
L. P. olivaceus fish were alterations and oxida-
stocked in each tank, tive stress in juvenile P.
with a total of 10 fish per olivaceus fish in a biofloc
tank. This configuration system
led to a total of 120 fish
being stocked across the
12 tanks. The fish were
grouped into six water
temperature categories,
with duplicates for each
category
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

Farfantepenaeus brasiliensis 0.015 WT (21, 24, 27, 30, 33 °C) A 30-day nursery phase F. brasiliensis shrimp Hostins et al. 2015
trial was conducted at nursed at 27 °C in a
different temperatures. F. super-intensive biofloc
brasiliensis (PL23) were dominated system dem-
stocked in each of the 25 onstrated superior growth
tanks, with a density of performance, including
600 shrimps/m2. Each tank increased productivity,
had a volume of 150 L quicker growth, and higher
survival rates
P. vannamei 0.004 WT (24, 28, and 32 °C) The experimental setup The optimal conditions for Ponce-Palafox et al. 2019
Salinity (10, 20, and 30 g/L) involved 27 indoor plastic achieving the highest spe-
Aquaculture International (2024) 32:9321–9374

tanks, each with a total cific surface area of FW,

capacity of 500 L, but only SGR, productivity, and
200 L were used. PL14 survival were determined
were grown in a nursery at to be a temperature of
a density of 2000 PL per 27.25 °C and a salinity of
cubic meter in BFT for a 25.5 g/L
duration of 4 weeks
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13
 Table 3  (continued)
9336

Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
P. vannamei WT (°C), TSS The BFT systems were The bioflocs have a richer Pimentel et al. 2020
operated using molasses elemental composition
or white sugar as a carbon in terms of nitrogen and
source, creating a favora- phosphorus. Systems with
ble heterotrophic environ- higher WT and higher
ment with C:N ratios all TSS have a greater ability
very close to 10 to absorb and immobilize
these elements in the
bioflocs
WT (15, 20, 25, and 30 °C) Each temperature group con- The study concluded that Yao et al. 2013
tained three replicates in a biofloc bacteria can
randomized block design. enhance the effectiveness
To ensure sufficient mix- of sludge treatment at
ing intensity and saturated 25 °C. At this temperature,
DO, a bubbling diffusive a significant amount of
air stone was used in ammonia nitrogen removal
each reactor. Glucose was was achieved in just 3 h
added to achieve a C:N without increasing the
ratio greater than 20 nitrite and nitrate concen-
trations in the SBRs. The
process was found to be
more efficient at 25 °C
than in the other three
temperature groups
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

O. niloticus 3.68 NaHCO3 The fish were distributed The recommendation Martins et al. 2017
CaCO3 into experimental units, of using ­NaHCO3 and
Ca(OH)2 with each unit having a Ca(OH)2 for alkalinity
for alkalinity and pH cor- useful volume of 37.5 and pH correction in Nile
rection L. There were 25 fish tilapia nursery in BFT sys-
per tank. When the TAN tems. These practices can
levels in the tanks reached enhance fish growth and
a value higher than net yield while potentially
1.0 mg/L, molasses was providing economic ben-
added to correct the C:N efits through the judicious
ratio. The C:N ratio was use of these compounds
adjusted to a ratio of 15:1
Aquaculture International (2024) 32:9321–9374

by adding molasses
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13
 Table 3  (continued)
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Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
P. vannamei 6 Alkalinity and suit- The experiment was con- It is evident that calcium Furtado et al. 2011
able sources (Ca(OH)2, ducted using juveniles hydroxide and sodium
­Na2CO3, ­NaHCO3) stocked in 150-L 12 tanks bicarbonate are effective
at a density of 333 shrimp/ compounds for increasing
m3 alkalinity in BFT cultures.
These compounds success-
fully elevated alkalinity
levels, which is important
for maintaining stable
water conditions and sup-
porting optimal fish health
and growth. The study
also revealed that main-
taining adequate levels of
alkalinity and pH is cru-
cial for water quality and
zootechnical performance
in BFT cultures. When
alkalinity concentrations
remained below 100 mg
­CaCO3/L and pH levels
were consistently below 7
for extended periods, both
water quality and the per-
formance of the fish were
negatively affected
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

Artemia franciscana Alkalinity: A total of 35 pairs of In this study, an alkalinity Naserizadeh et al. 2020
75, 150, 225, and 300 mg Artemia (male and female) level of 150 mg per liter
­CaCo3/L were cultured in 50-mL of calcium carbonate was
tubes under biofloc found to significantly
conditions for a period of improve the performance
3 weeks of biofloc production com-
pared to other alkalinity
levels. It is recommended
for the production of
biofloc for Artemia feed-
ing from the nauplii to the
adult stage
Aquaculture International (2024) 32:9321–9374

P. vannamei 0.2 Alkalinity: 12 experimental units, each Bioflocs were successfully Furtado et al. 2015
75, 150, 225, and 300 mg with a total volume of formed during the rearing
­Caco3/L 50 L, were filled with of P. vannamei without
30 juvenile shrimp. This the need for water renewal
stocking density resulted when the alkalinity levels
in a density of 150 exceeded 70 mg C ­ aCO3/L
shrimps/m2. The shrimp and pH levels were above
were kept in these units for 7.38. The treatments
a duration of 49 days as with higher alkalinity
part of the experiment levels, specifically at 300
­mgCaCO3/L, exhibited
favorable nitrification
rates and zootechnical
performance
9339

13
 Table 3  (continued)
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Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
O. niloticus 3.68 pH: 6.5, 7.5, 8.3 The experiment took place Maintaining a pH range of Martins et al. 2019
over a period of 60 days in 6.5–7.5 in biofloc systems
indoor tanks. Each experi- can lead to improved
mental unit, with a volume growth, higher net yield,
of 37.5 L, was stocked and reduced FCR in tilapia
with 25 fish. Liquid sugar nursery production. This
cane molasses was used to pH range enhances the
correct the C:N ratio, with efficiency of the biofloc
a ratio of 15:1 system for tilapia rearing
P. vannamei 6.56 pH: 7.1–7.6; 7.6–8.1 The experiment was car- The study’s findings indicate Zhang et al. 2017
ried out in nine indoor that adjusting the pH of
fiberglass tanks, each the culture water in BFT
with a water volume of systems from 7.6 to 8.1
90 L. The shrimp were yielded better results
transferred to these tanks compared to adjusting
at a density of 30 shrimp it from 7.1 to 7.6. These
per tank and were kept results will be valuable for
there for a period of shrimp producers aiming
48 days. The BFT method to maintain appropriate
was employed, and sodium pH and alkalinity levels in
bicarbonate was used to their BFT systems
adjust the pH levels
P. vannamei 0.24 Salinity: 2, 4, and 25 g/L The shrimp were grown at The highest growth perfor- Maica et al., 2012
a density of 300/m2, with mance and survival rates
molasses and wheat bran were seen at a salinity
serving as carbon sources. level of 25 g/L. However,
The C:N ratio was about it was also found that the
20:1, and the entire pro- shrimp could be success-
cess took 40 days fully raised at a salinity
level of 4 g/L
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

P. vannamei 1.79–1.89 Salinity: 9, 18, and 36 Pacific white shrimp were Salinity affects natural com- Decamp et al. 2003
stocked at 50 individuals/ munities and production of
m2 for 60 days P. vannamei
P. vannamei 2.56 Salinity: 10 and 32 g/L This research was conducted The trial found that P. van- Khanjani et al. 2020a
in indoor 300-L fiberglass namei exhibited superior
tanks, with each tank hold- performance in their
ing a total water volume juvenile stage when an
of 180 L. Each tank artificial diet was sup-
was populated with 70 plemented with biofloc at
individuals, corresponding a salinity level of 32
to a biomass of 1 g/L. To
Aquaculture International (2024) 32:9321–9374

stimulate the development

of bioflocs and maintain
an optimal C:N ratio of
15:1
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13
 Table 3  (continued)
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Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
Litopenaeus setiferus 0.1 Salinity: 5 and 35 g/L Juvenile shrimps weighing The biofloc system has been Valenzuela-Jiménez et al.
0.1 g were distributed found to be effective in 2022
across six fiberglass tanks, stabilizing and enhancing
with each tank housing conditions for L. setiferus,
3000 juveniles (equivalent a highly potential penaeid
to 150 shrimps/m3). These species for farming, under
juveniles were allowed to high salinity conditions.
acclimatize for five days. However, during the
Three groups were set up experimental phase, the
and monitored over the system was designed to
course of the experiment, stimulate oxidative stress,
which lasted 90 days. leading to increased cellu-
During the experimental lar damage at low salinity
period, molasses was levels due to high oxygen
added in a 6:1 C:N ratio consumption under fasting
whenever the Ammonia and postprandial condi-
level exceeded 1 mg per tions
liter
Red tilapia 13.78 Salinity: 5, 10, 15, and Red tilapia fry were stocked The current research Kumari et al. 2021
20 g/L in each biofloc unit at a indicates that when reared
rate of 150 fish per cubic in higher salinity condi-
meter. Jaggery was used as tions using inland saline
the carbon source to main- groundwater, red tilapia
tain a C/N ratio of 15:1 demonstrated improved
growth efficiency, survival
rates, biochemical indica-
tors, and carcass quality
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

P. vannamei Salinity: 10, 20, and 30 g/L P. vannamei (PL8) were Operating at a low salinity Ray and Lotz 2017
stocked at 4000 shrimp/m3 of 10‰ can significantly
for 54 days reduce salt usage by
approximately 50% com-
pared to a 20‰ treatment,
resulting in substantial
cost savings for production
facilities. This research
contributes to demon-
strating the versatility of
salinity options for shrimp
production in commercial-
Aquaculture International (2024) 32:9321–9374

scale biofloc systems

P. vannamei 74.46 mg Salinity: 8, 21, and 32 g/L Shrimps were cultivated in The study concludes that a Khanjani et al. 2020b
300-L fiberglass tanks, high salinity level of 32
each filled with 130 L ppt enhances the growth
of water, achieving a and survival of the biofloc
density of 1 post-larva supplemented Pacific
per liter. This culture was white shrimp in a BFT
maintained for a period of system
28 days
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13
 Table 3  (continued)
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Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
O. niloticus 1.75 Salinity: 0, 4, and 8 g/L Fingerling fish were reared The study suggests that Khanjani and Alizadeh 2024
for 37 days in tanks, each salinity levels of 4 and
having a water volume of 8 g/L can enhance water
140 L. The stocking den- quality, growth perfor-
sity was one fish per liter mance, survival, and
carcass composition of
Nile tilapia fingerlings in a
biofloc system. This find-
ing implies that brackish
waters can be effectively
used for sustainable
aquaculture in the rearing
of Nile tilapia
O. niloticus 93.8 Salinity: 0, 4, 8, 12, and Tilapia were cultivated for The study suggests that de Souza et al. 2019
16 g/L 90 days in fiberglass tanks, BFT, developed at
each with a volume of 800 salinities ranging from 0
L. The stocking density to 16 g/L, is suitable for
was set at 30 fish/m3 the culture of tilapia. The
water quality was main-
tained in proper conditions
for fish performance, with
high rates of survival
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

O. niloticus 40 Salinity: 0, 4, 8, 12, and The experiment was The study found that nitrite de Alvarenga et al. 2018
16 g/L conducted for 70 days in peaks occurred later and
150-L fiberglass tanks, were higher in salinities of
with 60 fish being kept in 12 and 16 g/L compared
each tank to other treatments.
Water that was salinized,
particularly within the
range of 4 to 8 g/L, can be
recommended for use in
a biofloc tank to enhance
the growth performance
of Nile tilapia during the
Aquaculture International (2024) 32:9321–9374

initial culture phase

O. niloticus 15.62 Salinity: 0, 10, and 20 g/L The experiment was The variations in salin- Luo et al. 2017
conducted for 150 days in ity levels in the Biofloc
tanks with a volume of 0.6 technology (BFT) system
­m3 and a density of 2.6 kg/ resulted in a notable dif-
m3 of fish. Glucose was ference in the abundance
added to the tanks at a rate of microorganisms at the
of 50% of the feed level genus level. However, the
to ensure the ideal C/N bacteria genus Cetobac-
ratio for the progress of terium was present in all
the BFT three groups
P. indicus 0.11 Salinity: 5‰, 15‰, 25‰, P. indicus, PL in the BFT The lower salinity rearing Das et al. 2023
30‰, and 35‰ with C:N ratio of 15:1 with biofloc did not show
during nursery phase in significantly lower growth
different salinities, for than optimum range
40 days of salinity in case of P.
P. indicus were stocked at indicus PL
the density of (1.2 PL/L)
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13
 Table 3  (continued)
9346

Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
Lates calcarifer 11 DO Asian sea bass (at a density The study successfully Whangchai et al. 2022
of 30 fish/m3) were established the use of
raised in 50-ton ponds for aeration devices and opti-
100 days mized feeding frequen-
cies to enhance oxygen
levels in 60-ton freshwater
recirculating aquaculture
ponds and the biofloc
system for Asian seabass
rearing. This approach
proved to be more efficient
than using an ejector, as
it increased dissolved
oxygen levels through the
use of small bubbles and a
large specific contact area
Piaractus mesopotamicus 20.70 TSS levels (0, 1500, 3000, The treatments were main- Pacu juveniles exhib- Pellegrin et al. 2022
4000, 5000, 6000, and tained in a static system ited tolerance to TSS
7000 mg/L) using 8 L tanks, with 6 concentrations of up to
L of usable volume, and 5000 mg/L, which are not
aeration. Only the water typically utilized in BFT
collected for water quality systems. The LC50-96 h
analysis was replaced. (lethal concentration that
Molasses, a carbon-rich causes mortality in 50%
substance, was used to of the population within
adjust the carbon to nitro- 96 h) of TSS for pacu was
gen ratio to 15:1 estimated to be 5477 mg/L
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

P. mesopotamicus 28.41 TSS levels (0, 250, 500, 750, 80-L tanks with a useful vol- The exposure of fish to Pellegrin et al. 2022
and 1000 mg/L) ume of 60 L were utilized. elevated TSS concentra-
Each tank contained six tions, particularly 750 and
fish, resulting in a total of 1000 mg/L, resulted in
18 fish per treatment. The increased levels of blood
duration of the experiment glucose, pH, erythrocytes,
was 5 days hemoglobin values, and
hematimetric indices on
the first day. However, by
the fifth day, most of these
parameters stabilized at
varying TSS concentra-
Aquaculture International (2024) 32:9321–9374

tions. It is important to
note that TSS concentra-
tions of up to 250 mg/L
are recommended for BFT
system production
9347

13
 Table 3  (continued)
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Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
Rhamdia quelen TSS: A 21-day trial was The control treatment in Poli et al. 2015
in three concentrations conducted to assess the the experiment recorded
up to 200, effectiveness of biofloc the lowest survival rate
400–600, technology in the growth (10.2%) due to an infesta-
800–1000 mg/L of R. quelen. The fish tion of Ichthyophthirius
were reared at a density multifiliis on the 15th day.
of 25 larvae per liter of However, no infestation
water. The purpose of the was observed in the bio-
experiment was to evalu- floc treatments, resulting
ate the performance and in a significantly higher
potential benefits of using mean survival rate (46.9%)
BFT in the production of compared to the control
R. quelen The study suggests that
R. quelen larvae can be
successfully grown in a
biofloc system with TSS
concentrations of up to
1000 mg/L. However, it
was found that the best
growth occurred in tanks
with a higher percent-
age of volatile suspended
solids (up to 200 mg/L)
and clear water achieved
through the daily addition
of dextrose
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

P. vannamei 6.8 TSS: Juvenile shrimp were According to the study, Schveitzer et al. 2013
in three concentrations: stocked in twelve 850-L intermediate levels of
200, tanks at a stocking density bioflocs, specifically TSS
400–600, of 459 shrimps/m3. The concentrations rang-
800–1000 mg/L study spanned a period ing between 400 and
of 44 days, during which 600 mg/L, were found to
the growth and develop- be more suitable for the
ment of the shrimp were super-intensive culture
monitored and evaluated. of P. vannamei shrimp.
The purpose of the study This finding suggests
was likely to assess the that maintaining TSS
performance and produc- concentrations between
Aquaculture International (2024) 32:9321–9374

tivity of the shrimp at the 400 and 600 mg/L can be

given stocking density and beneficial in supporting
tank size over the 44-day the successful super-
period intensive culture of P.
vannamei shrimp
P. vannamei 0.72 Low solids and high solids Eight raceways with a The study suggests that the Ray et al. 2011
volume of 50 ­m3 each management of biofloc
were randomly assigned concentration can have an
to one of two treatments: impact on nitrogen cycling
T-LS and T-HS. The T-LS pathways and shrimp
treatment had settling production. This indicates
chambers adjacent to the that controlling the level
raceways with a volume of biofloc concentration in
of 1700 L and a flow rate the system can influence
of 20 L/min. On the other the efficiency of nitrogen
hand, the T-HS treatment removal and utilization,
had settling chambers with which in turn can affect
a volume of 760 L and a the overall productivity of
flow rate of 10 L/min shrimp
9349

13
 Table 3  (continued)
9350

Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
P. vannamei Nauplius (N5) TSS, 300 and 600 mg/L In the 14-day study, the As the TSS levels increased, Liu et al. 2022
focus was on investigating there was an increase in
the nursery phase of the nitrate and total nitrogen
nauplius (N5) stage of P. concentrations. On the
vannamei in a BFT system other hand, alkalinity
and pH decreased with
increasing TSS levels.
Based on the findings, it
suggests that a BFT sys-
tem with TSS levels below
300 mg/L can achieve
better performance in
the nursery phase of P.
vannamei nauplii. This
indicates that maintaining
a lower TSS level in the
system can have a positive
impact on the overall
water quality and the
development of the nauplii
Aquaculture International (2024) 32:9321–9374
 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

Carassius auratus gibelio 6.4 TSS concentrations (10, 300, A tank with a volume of The optimal TSS concentra- Zhang et al. 2018
600, 800, and 1000 mg/L) 1.80 ­m3 was filled with tion for gibel carp culture
freshwater. The tank was was found to be within the
stocked with gibel carp at range of 600–800 mg/L.
a density of 250 individu- This suggests that main-
als. The experiment lasted taining TSS levels within
for a duration of 30 days. this range can promote
Molasses was added to the better growth and perfor-
tank 2 h after each feeding. mance of gibel carp in the
The molasses addition was system. Furthermore, the
done at a C/N ratio of 15 study results indicated that
bioflocs produced in the
Aquaculture International (2024) 32:9321–9374

BFT system had the abil-

ity to enhance the innate
immunity of gibel carp
Piaractus brachypomus 2.7 TSS, 200–300 and Fish fingerlings were The findings suggest Angeles-Escobar et al. 2022
400–600 mg/L exposed to different TSS that maintaining a TSS
concentrations in a biofloc concentration between
system for a period of 200 and 300 mg/L in
45 days. The TSS concen- biofloc systems is optimal
trations were likely varied for cultivating red pacu
to assess their effects on fingerlings without com-
fish growth and perfor- promising their health and
mance growth. This information
can be valuable for fish
farmers and aquaculture
practitioners in optimizing
biofloc system conditions
for red pacu fingerling
production
9351

13
 Table 3  (continued)
9352

Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
O. niloticus, Sciaenops ocel- 85 tilapia, 2 TSS The experiment compared The study suggests that Poersch et al. 2021
latus, and P. vannamei shrimp, and 71 the use of tilapia to control Pacific white shrimp, red
red drum suspended solids (biologi- drum, and tilapia can be
cal treatment) to control successfully cultured in
systems without tilapia. an integrated multispecies
The initial mean weights system using BFT. This
and stocking densities indicates that these three
were: 2 g and 250 ind/ species can coexist and
m3 for shrimp; 71 g and thrive in the same system,
1.04 kg/m3 for red drum; which can have potential
3
and 85 g and 5.12 kg/m benefits such as resource
for tilapia utilization and waste
management
P. vannamei 4.54 TSS In the 42-day trial, different The findings emphasize the Gaona et al. 2017
100–300, 300–600, and treatments with three importance of carefully
600–1000 mg/L ranges of TSS were tested. managing TSS concentra-
Shrimp juveniles were tions (between 100 and
stocked at a density of 372 300 mg/L) during the for-
shrimps/m3 mation and maintenance
of biofloc systems. By
maintaining TSS within
the recommended range
and reducing variations,
aquaculture practition-
ers can optimize water
quality, enhance nutrient
recycling, and support the
overall health and per-
formance of the cultured
organisms
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 Table 3  (continued)
Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

P. vannamei 6.2 TSS The experimental design Cultivation of U. lactuca in Carvalho et al. 2023
30 and 400 mg/L was carried out with shrimp effluent water with
three treatments, (1) a TSS level of 30 mg/L
BFT: cultivation of Ulva resulted in significantly
lactuca in effluent from lower TSS levels com-
shrimp cultivation in a pared to the BFT groups,
BFT, with 400 mg/L of which had a TSS level of
TSS; (2) DEC: cultivation 400 mg/L. This method
of U. lactuca in effluent can be used to improve
from shrimp culture in water quality and reduce
a BFT, after a period of environmental impacts
decantation of solids, with associated with shrimp
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30 mg/L of TSS; (3) VS: farming

cultivation of U. lactuca
in standard von Stosch
enrichment solution at a
concentration of 10 mL/L,
for 35 days
O. niloticus and Samhong Heavy metals: Cu, Fe, and Fish ponds were filled with The concentrations of Cu Deswati et al. 2022
mustard (Brassica juncea Zn 0.88 ­m3 of water, and (copper), Fe (iron), and
L.) 1000 fish with a length of Zn (zinc) in the fish ponds
2–3 cm were added. The were within the ranges of
rearing period for the fish 0.0036–0.02260 mg/L,
was 42 days 0.5269–1.1013 mg/L, and
0.0215–0.2581 mg/L,
respectively. While only
the Cu concentrations did
not exceed the quality
standard, the growth of
O. niloticus and B. juncea
was fast, and fish survival
was high (> 95%)
9353

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 Table 3  (continued)
9354

Cultured species IBW (g) Influential parameter Relevant rearing conditions Main findings Reference

13
O. niloticus 8.32 MPs Nile tilapia were exposed to It can be concluded that Hu et al. 2023
MPs made of polystyrene MPs had no significant
with a diameter range of effects on nitrification in
32–40 μm. The exposure the BFT systems or on
was conducted in a BFT the digestive activity of
system for a duration of tilapia. The presence of
28 days. Three different MPs did not significantly
treatment groups were influence the microbial
used: 0 items/L (con- communities in the BFT
trol group), 80 items/L systems or the gut micro-
(equivalent to 30 μg/L), biota of the tilapia
and 800 items/L (equiva-
lent to 300 μg/L)

Abbreviations: IBW initial body weight, WT water temperature, C:N carbon to nitrogen ratio, GIFT genetically improved farmed tilapia, DO dissolved oxygen, TSS total sus-
pended solids, T-LS treatment-low solids, T-HS treatment-high solids, ROS reactive oxygen species, FW final weight, SGR specific growth rate, SBRs sequencing batch reac-
tors, PL postlarvae, PPT parts per thousand, MPs microplastics
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Aquaculture International (2024) 32:9321–9374 9355

Temperature

Water temperature is a key determinant of the health and survival of cultured organ-
isms as fluctuations in temperature can adversely affect growth and survival (Whang-
chai et al. 2018; Cheng et al. 2018). Temperature also influences various chemical and
biological parameters, including DO and the rate of chemical reactions (Mubiana and
Blust 2007; Hostins et al. 2015). Increasing water temperatures can result in a drop in
DO, leading to respiratory distress marked by difficult breathing and heightened oxy-
gen exchange on the surface of the gills (Lewis et al. 2012). Temperature also affects
physiological pathways, behaviors, and reproduction, leading to changes in maturation
and stages of reproduction such as ovulation, hatching, and embryogenesis (Narum
et al. 2013; Kim et al. 2017). Microbial metabolism is similarly affected by tempera-
ture (Price and Sowers 2004). Very cold temperatures (4 °C) promote deflocculation
compared to cool temperatures (18–20 °C), likely due to reduced microbial activity
in the flocs (Wilen et al. 2000). In warmer waters (30–35 °C), sludge bulking occurs,
characterized by high sludge volume index (SVI ≥ 500 mL/g), due to overproduction of
extracellular polymeric substances (Krishna and Van Loosdrecht 1999). At moderate
temperatures (20–25 °C), flocs are more stable with a medium SVI of around 200 mL/g.
It can be challenging to adjust water temperature in BFT ponds, particularly in outdoor
farms, due to the costs associated with temperature regulation (Phulia et al. 2012). Tem-
perature greatly impacts biofloc characteristics. Lower temperatures restrict biological
processes including the activity of microbial organisms (Arnosti et al. 1998; Garcia
et al. 2011; de Souza et al. 2016). This could impede the growth of biofloc microorgan-
isms and lower the rate of nitrification. This effect is most pronounced when the cold
exposure is sudden. Experiences from activated sludge systems suggest suitable organic
matter biodegradation and nitrification under sub-20 °C conditions, but extremely low
temperatures may potentially lead to a complete cessation of nitrification (Johnston
et al. 2019). The mechanisms underpinning the relationship between temperature and
nitrification are still unclear, but changes in the interactions between microorganisms
are one of the possibilities. An overwintering experiment in tilapia culture showed that
existing bioflocs withstood < 4 °C temperatures and still carried out nitrogen removal
(Crab et al. 2009). Water temperature is also critical for white shrimp P. vannamei.
Extreme temperature fluctuations might lower immune resistance, increasing vulner-
ability to Vibrio infection (Cheng et al. 2005). Since many viral infections in shrimp are
affected by temperature, it is advisable to maintain water temperature at around 29 °C or
higher, especially to control viruses involved in white spot syndrome (Lim et al. 2021).
In BFT, the optimal temperature range for shrimp is around 30 °C (27–33 °C for giant
freshwater prawn, 28–30 °C for Pacific white shrimp, and 28–32 °C for the giant tiger
prawn). However, shrimp can endure lower temperatures at the cost of growth. A study
by de Souza et al. (2016) found the best results at 27 °C, with shrimp showing superior
growth and reduced oxidative stress. Tilapia can survive in cold conditions (< 5 °C), but
will not feed at < 15 °C (Crab et al. 2009). Similar temperature preferences are observed
in African catfish (Clarias gariepinus), channel catfish (Ictalurus punctatus), and milk-
fish (Chanos chanos). On the other hand, species like gray mullet (Mugil cephalus) and
common carp (Cyprinus carpio) thrive below 30 °C, but require temperatures above
20 °C (Bagi et al. 2023).
Tables 2 and 3 provide detailed information on the suitable temperature ranges for
shrimp and tilapia, and the impact of varying water temperatures on biofloc system

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performance and the cultivated species. A study by Ponce-Palafox et al. (2019) observed
a decrease in pH with rising temperatures in BFT shrimp aquaculture, particularly
at 32 °C, mirroring results from similar research in the past (Hostins et al. 2015; de
Souza et al. 2016). Additionally, at 32 °C, there tends to be an increase in TAN, nitrites,
nitrates, TSS, and SS, aligning with findings from the literature (de Souza et al. 2014;
Hostins et al. 2015).

pH and alkalinity

Maintaining proper pH is crucial in BFT (Martins et al. 2019). It is recommended to main-

tain pH within the 7.0–8.5 range to ensure the desired functioning of the system (Emeren-
ciano et al. 2017). Even minor changes in pH can significantly impact the biochemical pro-
cesses in water (Suchithra et al. 2016; Ahammed et al. 2022). For instance, marine shrimp
culture exhibits optimal growth in neutral to mildly basic environments (pH = 7.0–9.0)
(Van Wyk and Scarpa 1999). pH is a determinant in nearly all chemical reactions, impact-
ing both water chemistry and the physiology of shrimp (Lemonnier et al. 2004). pH < 6.5
or > 9.5 has been linked to inhibited growth in Penaeidae species. The optimal pH range for
shrimp culture (7.0–9.0) is also conducive to the proliferation of heterotrophic and nitrify-
ing microorganisms (Chen et al. 2006; Van Wyk and Scarpa 1999).
Previous research has indicated that the downward pH trend observed in biofloc systems
may be attributable to bacterial production (Azim et al. 2008; Lim et al. 2021), increased
respiration by heterotrophic microbes, and elevated C ­ O2 levels (Emerenciano et al. 2012;
Miao et al. 2017; Vinatea et al. 2010; Gaona et al. 2016). McIntosh (2001) highlighted that
higher densities lead to increased respiratory rates and elevated C ­ O2 levels, subsequently
causing pH reduction. A study conducted in Belize showed that P. vannamei exhibited
resilience to pH levels between 6.8 and 7.3. Conversely, Wasielesky et al. (2006b) report
that pH < 7 adversely affects the growth of P. vannamei in BFT culture. Hence, it is essen-
tial to conduct experimental studies regarding the impact of alkalizers on the quality of
water and the performance of P. vannamei.
pH has a significant influence on bacterial growth, with the optimal range for hetero-
trophic bacteria being between 7.0 and 10.0 (Mayo and Noike 1996); the figure for auto-
trophic species is 7.0–8.0 (Chen et al. 2006). Factors such as inorganic nitrogen levels, total
biomass of bacteria, DO, pH, and temperature are known to influence the rate of nitrifica-
tion (Chen et al. 2006). The role of alkalinity in nitrification is also significant, with studies
highlighting the substantial influence of inorganic carbon levels, particularly in the produc-
tion of nitrite from ammonia (Chen et al. 2012). Several studies have assessed the effect of
pH on nitrification rate in the context of bioreactors and effluent treatment. Martins et al.
(2019) observed no significant pH-dependent effects on the initial stage of nitrification, as
the total conversion of ammonia to nitrite remained consistent across different treatments.
However, the subsequent phase displayed a correlation with pH, particularly regarding the
rate of nitrification. The maximum nitrite-nitrogen concentration occurred at pH = 6.5.
Various experiments, such as the study by Jiménez et al. (2011), have demonstrated robust
nitrification at pH = 7.5, and a cessation of the process at pH = 6.5. In contrast, Tarre and
Green (2004) observed active nitrification at pH = 4.3 in a bioreactor. Villano et al. (2010)
illustrated that pH is fundamental to the composition of the biofloc microbial community,
promoting adaptability to environmental shifts and the regulation of nutrient uptake and
metabolism (Lauro et al. 2009). The research by Martins et al. (2019) further indicates

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the feasibility of sustaining nitrification at pH = 6.5 through managing the composition of

microbial flora.
Alkalinity is defined as the capacity to neutralize acids and maintain pH stability (Fur-
tado et al. 2015). Biofloc culture water tends to decrease pH over time, so demand for
alkalizing agents increases (Ebeling et al. 2006b; Lim et al. 2021). Given that pH values
outside the 7.0–8.0 range can significantly affect biochemical reactions and physiologi-
cal processes, dolomite must be steadily supplied to restore and maintain water alkalin-
ity. There are no definitive guidelines regarding maximum alkalinity levels, but alkalinity
should be maintained above 100 mg C ­ aCO3/L, especially in salt water culture conditions
(Crab et al. 2009; Furtado et al. 2011). Higher alkalinity levels (up to 300 mg ­CaCO3/L)
have been associated with improved nitrification and shrimp growth (Furtado et al. 2015).
Van Wyk and Scarpa (1999) state that P. vannamei requires alkalinity > 100 mg ­CaCO3/L
for optimal shrimp growth, also noting that higher alkalinity levels can reduce daily pH
fluctuations. In a BFT system for P. vannamei culture with a density of < 610 shrimp/m2,
Vinatea et al. (2010) utilized 0.12 g/L pure ­NaHCO3 to elevate alkalinity when values were
below 60 mg C ­ aCO3/L. In freshwater conditions, studies on Nile tilapia in BFT systems
have presented the consumption of alkalizing agents and decreased pH (Alvarenga et al.
2018; Azim & Little 2008; Pérez‐Fuentes et al. 2016). In BFT systems, pH can decrease
due to decreased alkalinity, elevated C ­ O2, and increased respiration (Vinatea et al. 2010).
Therefore, it is essential to maintain proper alkalinity levels via supplementation of alkaliz-
ing agents such as sodium carbonate, calcium hydroxide, and sodium bicarbonate before
and during the production cycles (Boyd et al. 2016).
Maintaining pH in the 7.5–7.9 range and an alkalinity of 100–162.2 mg ­CaCO3/L in
BFT systems remarkably improves productivity (Furtado et al. 2011). Furthermore, when
alkalinity is maintained in the 40–160 mg C ­ aCO3/L range, TSS, ammonium, nitrite, and
nitrate are unaffected (Piérri 2012). However, alkalinity > 160 mg ­ CaCO3/L has been
linked to increased pH, boost biomass of heterotrophic and ammonium-oxidizing bacteria,
and a decrease in nitrite-oxidizing bacteria (Piérri 2012). Water alkalinity and pH are also
affected by the solids management (Schveitzer et al. 2013). These authors found that the
amount of alkalinity and pH in P. vannamei biofloc water is higher with 200–400 mg/L
of TSS when compared to higher levels (400 to 600 mg/L, and 800 to 1000 mg/L). This
higher level of alkalinity in 200–400 mg/L of TSS could be a response to denitrification as
pointed out in past studies (Ebeling et al. 2006a; Ray et al. 2010a, b, 2011).

Salinity

Salinity range can play a major role in biofloc development and impact the microbial com-
munity (Maicá et al. 2014). Table 2 shows the most common values of salinity in BFT,
where its optimal levels are species-specific. Table 3 shows the effects of different salini-
ties on the characteristics of BFT systems and cultured species. For instance, Hakanson
(2006) observed a trend for suspended particle aggregation and floc size to increase with
higher salinity. Studies conducted by Maicá et al. (2012) and Khanjani et al. (2020a) with
P. vannamei indicate a progressive increase in biofloc levels (settling solids), reaching its
peak at elevated salinity levels. The same trend was observed by Decamp et al. (2003) also
highlighting a decrease in pH at higher salinity (36 g/L) in comparison to lower salinity
treatments (18 and 9 g/L) in zero-water-exchange P. vannamei juvenile culture. According
to the authors, this phenomenon is attributed to enhanced photosynthetic activity at lower
salinities. This trend was also noted by Maicá et al. (2012) with P. vannamei biofloc-based

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culture where higher biofloc levels were observed at 25 g/L, leading to lower pH as a result
of high microbial respiration (Wasielesky et al. 2006a).
Several studies also highlighted that salinity changes in water can also impact the meat
nutritional quality (Huang et al. 2004; Khanjani et al. 2020a; 2020b).
In BFT, Khanjani et al. (2020a) demonstrated an increase in dry matter, protein, lipid,
and ash in shrimp corroborated with rising salinity. The relationship between protein con-
tent and salinity was further investigated by Liang et al. (2008) and Maicá et al. (2014).
Maicá et al. (2014) reported higher ash and lipid content in shrimp at higher salinities,
emphasizing the influence of biofloc-based culture on shrimp biochemistry, as indicated
by elevated ash and lipid levels in no-exchange treatments compared to controls. Khanjani
et al. (2020a) observed elevated biofloc protein content at higher salinity levels. However,
Ju et al. (2008) and Maicá et al. (2014) reported elevated protein levels at minimum salin-
ity. With the increase in salinity, a decrease in biofloc lipid content was noted, with the
highest and lowest levels respectively observed at 10 and 32 g/L (Khanjani et al. 2020a).
Biofloc ash content was also directly correlated with salinity (Maicá et al. 2014), the same
trend corroborated by Ju et al. (2008), who reported ash percentages of 18.3 and 40.7% at
5 and 32 g/L, respectively. Valenzuela-Jiménez et al. (2022) conducted a comprehensive
study on the effect of low (3 g/L) and high (35 g/L) salinities on oxygen consumption, per-
formance, oxidative stress, and antioxidant activity in outdoor BFT culture of Litopenaeus
setiferus. The study revealed improved feed utilization and FCR with higher salinity and
reduced weight gain and survival in low salinity. The poor performance in low salinity is
attributed to the unbalanced ionic composition and nitrite toxicity (Soares de Moura et al.
2021).

Dissolved oxygen (DO)

Suitable DO level is an important factor for proper development of bioflocs (De Schryver
et al. 2008). Studies indicated a correlation between DO and the growth and densification
of bioflocs (Wilen and Balmer 1999). Suboptimal settling characteristics in bioflocs were
found at reduced DO (0.5–2.0 mg/L), evidenced by an SVI of 250 mL/g. In contrast, a
higher DO (2.0–5.0 mg/L) reduces SVI to approximately 100 mL/g (Phulia et al. 2012).
However, it is possible for bioflocs with larger SVI to develop under diminished DO condi-
tions. For instance, these bioflocs can be consumed by the cultured species before the bio-
flocs settle to the bottom (Khanjani and Sharifinia 2024). Nonetheless, culture conditions
with high concentration of suspended particles (bioflocs) can obstruct the gill structures of
fish and shrimp (Emerenciano et al. 2013).
BFT with shared environment of cultured animals and microbials (bioflocs) can face
substantial oxygen demand (Yu et al. 2023a, b). Especially in intensive farming systems,
shrimp and fish typically require high DO levels, normally exceeding 5 mg/L (Patkaew
et al. 2024). A culture medium with high biofloc concentration can significantly elevate
the demand for oxygen, occasionally surpassing the requirements of cultured species them-
selves (Raza et al. 2024). In addition, the oxygen requirements can differ between in situ
and ex situ biofloc systems. The DO demand in in situ systems relates to the sum of cul-
tured species and bioflocs requirements, whereas in ex-situ systems where bioflocs are
formed externally in a decoupled biological reactor, the primarily oxygen demand relates to
the microbial communities (Ulloa-Walker et al., 2020). In this sense, in biological reactors,
a minimal DO requirement of 1 mg/L can address the needs of biofloc organisms (Kuhn

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and Lawrence 2012). For effective in situ and ex situ operations, the biofloc facility must
incorporate emergency power sources to counteract potential power outages, as inadequate
aeration can swiftly lead to critically low DO levels and mortalities (Vinatea et al. 2009).
Vinatea et al. (2009) found that in the absence of aeration, a DO concentration of 4.1 mg/L
could plummet to a critical level (0.65 mg/L) within 31.8 and 34.7 min in superintensive
(610 shrimp/m2) P. vannamei culture, depending on the presence or absence of light. Their
subsequent study (Vinatea et al. 2010) revealed a higher oxygen consumption rate in super-
intensive BFT shrimp culture under illuminated conditions up to 20 min after aeration was
stopped, at which point the dark condition had higher oxygen consumption. To enhance
DO and aeration efficiency in BFT systems, several aeration devices have been evaluated,
including paddle-wheels, venturis-nozzles, blowers with diffusers, submerged double helix
aerators, and mixed combinations (Pasco 2015; Piñeros-Roldan et al. 2020). The suitable
device or combination in BFT will depend on cultured species selection (e.g., shrimp or
fish), phase (nursery or grow-out), tank and pond design, and water depth, among other
characteristics (Piñeros-Roldan et al. 2020).

Total suspended solids (TSS) and settling solids (SS)

TSS and SS are examples of critical water quality parameters utilized for assessing bio-
floc concentrations in culture conditions (Soaudy et al. 2023). However, depending on the
method selected, accuracy and time consumption can vary drastically. This can be partially
solved as some studies reveal correlation among SS, TSS, water transparency, and turbid-
ity (Ray et al. 2010a, b; Schveitzer et al. 2013; Khanjani et al. 2021), meaning more cost-
effective ways to monitor the biofloc concentrations.
The optimal values of SS and TSS for biofloc systems are presented in Tables 2 and 3.
It is important to note that optimal TSS ranges can vary by species. For instance, Avni-
melech (2012) recommends 200–400 mg/L for tilapia. Hatem et al. (2013) recommend a
maximum TSS range of 220–250 mg/L for tilapia to avoid gill clogging, but higher levels
may be detrimental for earlier tilapia stages (or nursery phase) as highlighted by Emer-
enciano et al. (2017). In contrast, Poli et al. (2015) observed no significant variation in
Rhamdia quelen fish larvae survival across a TSS range of 200–1000 mg/L, indicating
the animals’ tolerance to high biofloc environments. Fingerling stage (5.7–10.2 g) of red
cachama (Piaractus brachypomus) tolerates TSS levels ranging from 384 to 645 mg/L
with survival rates between 84 and 99% (Ueno-Fukura et al. 2019). Similar levels were
found by Angeles-Escobar et al. (2022) for the same species with levels ranging from 200
to 600 mg/L; however, optimal performance was achieved at TSS = 200–300 mg/L. For
fish, species, phase of culture, and natural conditions in the wild seem to have impact on
the animal’s ability to tolerate high TSS levels in culture conditions. For marine shrimp,
the literature is more abundant regarding TSS and its impact on performance, water qual-
ity, and health. The most common range of TSS maintained in biofloc systems for shrimp
is between 400 and 600 mg/L (Avnimelech 2012; Gaona et al. 2011; Samocha et al. 2007;
Schveitzer et al. 2013). Higher TSS levels have been associated with negative performance
of P. vannamei (Vinatea et al. 2010; Schveitzer et al. 2013). Studies suggest that TSS levels
below 600 mg/L are most suitable for shrimp farming in biofloc systems (Ray et al. 2010a,
b; Schveitzer et al. 2013). Regarding P. vannamei nauplii, Liu et al. (2022) recommend
TSS level < 300 mg/L to achieve improved performance in larviculture. TSS can also have
a key impact on water quality parameters. Ray et al. (2011) found that at TSS = 200 mg/L,
nitrite and nitrate levels are lower while ammonium and phosphate levels are higher

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compared to TSS = 300 mg/L. In the same study, the growth rate was also higher (1.7 g/
week) at 200 mg/L than at 300 mg/L (1.3 g/week), and at TSS = 800–1000 mg/L, shrimp
experienced poor growth and significant gill obstruction. The authors suggested that lack
of suitable substrates for nitrifying bacteria, rapid denitrification in settling chambers, or
algal nitrate assimilation can have a severe impact on N-compounds fluctuations (Ray et al.
2011). In the same context, Schveitzer et al. (2013) recommend maintaining TSS between
200 and 600 mg/L for optimal growth and system stability. According to both studies, low
TSS conditions exhibited faster shrimp growth and enhanced environment conditions,
highlighting the role of biofloc concentrations on nitrogen cycling. In addition, maintaining
TSS in suitable conditions helps the functionality of biofloc systems, emphasizing the bal-
ance between bacterial oxygen demand and system productivity (Hargreaves 2013).
Throughout the culture cycle, TSS tends to increase if not properly managed (e.g., use
of clarifiers; Ray et al. 2011; Gaona et al. 2011). Higher TSS levels can decrease DO, pH,
alkalinity, and ammonia, but promoting nitrate accumulation (Avnimelech 2012; Gaona
et al. 2017). Furtado et al. (2011) also observed that changes in pH and alkalinity can
impact TSS levels. However, saturated culture conditions with high TSS levels may also
result in higher nitrite concentrations (Samocha et al. 2007; Luo et al. 2013). The addi-
tion of external organic carbon sources can help reduce ammonia levels (Lin and Chen
2001), but unlikely to control nitrite levels especially in high TSS conditions with DO, pH,
and alkalinity fluctuations. Suitable ranges of these parameters are critical to ensure proper
chemoautotrophic bacteria metabolism, especially nitrite-oxidizing bacteria (Ebeling et al.
2006b). Low levels of TSS may result in a more stable culture environment, avoiding trig-
gered competition for DO, micronutrients, and space by heterotrophic and chemoauto-
trophic (nitrifying) bacteria (Luo et al. 2013).
In terms of practicality, SS is considered a cost-effective way to measure solids concen-
tration in BFT, especially in commercial operations (Avnimelech 2012). Lim et al. (2021)
in a 70-day P. vannamei culture cycle reported insignificant SS in the first 30 days, fol-
lowed by a significant increase during days 35–70. Low SS levels have been suggested to
enhance the overall biofloc conditions and system stability (Lim et al. 2021).
The red-bellied pacu (Piaractus brachypomus) has demonstrated a notable toler-
ance to varying levels of SS. In juvenile stages (33.5–182.6 g), SS concentrations of
56.6 ± 20.7 mL/L were tolerated (Brú-Cordero et al. 2017). The tolerance of different SS
levels may be species-phase related, having impact on other water quality parameters, ani-
mal performance, and health.

Nitrogenous compounds

Nitrogen, an essential element for all living organisms, is supplied through dietary pro-
teins which are digested and used to synthesize various biomolecules (Wei et al. 2016).
Nitrogen accounts for approximately 5% of an organisms’ dry weight (Sigee 2005). Within
aquatic ecosystems, nitrogen exists in both organic and inorganic forms. Organic nitrogen
is primarily derived from residual feed, animal waste, dead matter, and urea. In contrast,
inorganic nitrogen encompasses ammonia and ammonium ­(NH3/NH4+), nitrite (­NO2−),
nitrate ­(NO3−), and to a lesser extent, nitrogen gas ­(N2) (Jiménez-Ojeda et al. 2018). In
aquatic environments, especially BFT conditions, heterotrophic bacteria and protozoa
are critical organisms capable of utilizing undissolved nitrogen (Sigee 2005) sourced by
organic matter continuously added to the systems. Understanding the different nitrogen
pathways, acting microorganisms and suitable compounds ranges are key for proper water

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quality maintenance and decision-making in BFT (Khanjani et al. 2022). Some of these
compounds can be highly toxic (e.g., N ­ H3 and ­NO2−), and poor water quality management
can lead to system collapse and high mortality rates (Lin et al. 2023). At high concentra-
tions, these compounds may impair oxygen transport and induce gill irritations and lesions,
as well as lead to hypoxia, enzymatic dysfunctions, and impair T4 hormone production
(Abakari et al. 2021).
Table 2 presents suitable nitrogen compound concentrations in biofloc systems for dif-
ferent species and Table 4 presents the microorganisms involved in the removal of nitrog-
enous compounds in BFT. The successful operation of BFT largely hinges on the effective
management of water quality, especially conversion of harmful nitrogenous compounds.
In a stable BFT environment, parameters such as ammonia and nitrite must be maintained
within acceptable ranges and continuously converted into benign compounds and biomass
through microbial processes (Khanjani et al. 2022). Ebeling et al. (2006a) and Khanjani
et al. (2022) present a stoichiometric framework for nitrogen removal in BFT while con-
sidering microbial growth and metabolic kinetics. The framework provides information
on substrate requirements, biomass production, and the outcomes of primary nitrogen
removal pathways, namely algal assimilation, autotrophic nitrification, and heterotrophic
assimilation.
Algal assimilation is often the predominant pathway for nitrogen removal in outdoor
BFT systems, with continuous light exposure (Zemor et al. 2019). On the other hand, in
conditions where photosynthetic activity is suppressed (e.g., using shading or indoor envi-
ronments), chemoautotrophic nitrification and heterotrophic assimilation are considered
the prevalent pathways for nitrogen removal (Avnimelech 2012; Samocha et al., 2017).
Algae assimilate C ­ O2 into organic carbon, and ammonia is directly incorporated into algal
biomass. Normally, the need for external organic carbon supplementation is reduced,
except to promote mixotrophic growth (Cecchin et al. 2018), and fluctuations of alkalinity
and pH are less common while nitrogen is assimilated during photosynthesis (Mohsenpour
et al. 2021). However, in these systems, there is a risk of algal population crashes, abruptly
releasing nitrogen back into the water (­ NO2 spikes) and causing sudden mortalities (Samo-
cha et al. 2017). In these systems, aeration and mixing are still needed to maintain optimal
DO.
In more controlled shaded or indoor BFT culture conditions, ammonia oxidation fol-
lowed by nitrite oxidation (nitrifying bacteria) and ammonia assimilation by hetero-
trophic bacteria were found as main pathways for toxic N-compounds control (Khanjani
et al. 2024c). In addition, denitrification, transforming nitrate into N­ 2 (Samocha 2019),
and integrated systems using plants (Pinho et al. 2022) are also presented as potential tool
to prevent N accumulation in BFT (Deng et al. 2020; Rajta et al. 2020). Nitrification via

Table 4  Microorganisms involved in the removal of nitrogenous compounds in the biofloc system (Ebeling
et al. 2006a; Jiménez-Ojeda et al. 2018; Khanjani et al. 2022)
Heterotrophic Chemoautotrophic Photoautotrophic

Main energy source Organic matter Inorganic compounds Solar radiation

Main nitrogen source NH4+ NH4+, ­NO2− NH4+, ­NO3−
Main carbon source Organic Inorganic Inorganic
Dominant process Assimilation Nitrification, conversion Photosynthesis, assimilation, removal
Relative growth rate Fast Slow Dependent on sunlight, fast

Abbreviations: NH4+ ammonium, NO2− nitrite, NO3− nitrate

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chemoautotrophic bacteria produces nitrate while minimal biomass is added to bioflocs due
to the slow growth of nitrifying microorganisms. In heterotrophic pathway, proper organic
carbon management is mandatory, both requiring substantial aeration and alkalinity/pH
control (Khanjani et al. 2024b). Collectively, these processes offer BFT operators various
strategies to manage harmful nitrogenous compounds (Leong et al. 2021; Roy et al. 2019).
Emerenciano et al. (2022a) bring a comprehensive review summarizing several intensive
penaeid systems including biofloc technology and similar microbial-based systems (e.g.,
Synbiotics, Aquamimicry, semi-biofloc). The authors highlighted the role of the different
microbial-based farming techniques, and its direct effect on microbial pathway and ulti-
mately nitrogen removal. Therefore, the selection and management of culture conditions
are directly linked to N-compounds management in BFT-based aquaculture.

Phosphate

Monitoring and maintaining phosphate ­(PO43−) levels in aquaculture systems is crucial for
recirculation management and effluent discharge. Due to its limited water exchange condi-
tion, in BFT, phosphate tends to accumulate over time; however, it can vary according to
the biofloc approach and culture conditions (Emerenciano et al. 2022b). Lim et al. (2021)
found a direct correlation between phosphate levels, time, and TSS in BFT. High (­PO43−)
levels may cause reduction in N:P ratio, leading to cyanobacteria outbreaks and less prev-
alence of desirable photosynthetic organisms such as chlorophytes and diatoms (Smith
1983). In a healthy and stable BFT, these groups should be prevalent in the phytoplank-
ton assemblage, known as key phosphate consumers. Unlike nitrogen, phosphorus does not
volatilize, so the forms present in water include organic forms and residual orthophosphate
(Silva et al. 2013).
Luo et al. (2014) observed lower phosphate concentrations in BFT systems compared
to RAS systems, suggesting active microbial phosphate cycling in bioflocs. Ray and Lotz
(2014) also reported reduced phosphate levels, attributing this to uptake by algae and het-
erotrophic bacteria, as corroborated by Longnecker et al. (2010) and Liu et al. (2018).
Feed input directly influences phosphate levels (Montoya et al. 2000). Feed and animal
waste are the primary sources of phosphorus in water (Barak et al. 2003). In BFT systems,
P. vannamei can incorporate up to 35% of the system’s total phosphorus intake, with the
remaining phosphorus existing in dissolved and particulate forms (Silva et al. 2013). How-
ever, fish and shrimp are normally resistant to phosphate concentrations, as shown by the
absence of reports of phosphate toxicity (Silva et al. 2013). Due to its limited literature,
there is a need for more R&D to better elucidate the phosphate role in BFT-based culture
conditions.

Heavy metals and microplastics: key considerations for BFT

Heavy metals (HMs) are a group of naturally occurring elements with high atomic weight
that are often toxic to humans and animals (Mitra et al. 2022). These elements can enter
aquaculture systems through various sources, including feed, water, sediment, and air
(Hossain et al. 2023a). Once in the system, HMs can accumulate in the tissues of fish and
other aquatic organisms, and may pose a health risk to consumers. There is little infor-
mation regarding BFT-based aquaculture and heavy metals. There is evidence that BFT
systems are capable of removing HMs from the culture environment (Deswati et al. 2022;
Habib et al. 2023a; 2023b). In O. niloticus culture, Habib et al. (2023b) found that BFT

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was effective in removing HMs (Cu, Cd, Zn, Pb, and Cr) with lower concentrations com-
pared to an earthen pond system. In an integrated tilapia (O. niloticus) and Samhong mus-
tard (Brassica juncea L.) biofloc-based culture, Deswati et al. (2022) observed a notable
reduction in Cu levels on day of culture (DOC) 42 onwards. The authors suggested that
biofloc-based aquaponics may mitigate metal concentrations via microbial activity and
plant absorption while preserving water quality (Deswati et al. 2022). In terms of micro-
bial community, cyanobacteria are a type of bacteria typically found in both freshwater
and marine BFT-based aquaculture (Khanjani et al. 2022). These microbes can produce
extracellular polymeric substances (EPS) able to foster particle aggregation (bioflocs),
bind to HMs, and act as a carrier throughout the system (Lama et al. 2016). The sus-
pended particles (bioflocs) can now be easily removed utilizing settling chambers (Ray
et al. 2011; Gaona et al. 2011). In a study by Bender et al. (1991 and 1994), the cyano-
bacterium Oscillatoria sp. was shown to be effective at removing a variety of HMs from
water, including Mn, Pb, Cd, Cu, Zn, Co, Cr, and Fe. The authors observed that HM lev-
els in water treated with Oscillatoria sp. was significantly lower than those in untreated
water.
As per HMs, there is also limited information regarding BFT-based aquaculture systems
and microplastics. Microplastics (MPs) are plastic particles < 5 mm and present a consider-
able challenge in the field of aquaculture. MPs have been observed in a range of aquacul-
ture settings, including aquaculture ponds (Ma et al. 2020), rice-fish co-culture systems (Lv
et al. 2019), estuarine aquaculture areas (Liu et al. 2023), RAS systems (Lu et al. 2019),
and BFT systems (Hu et al. 2023). In koi carp culture, accumulative MP uptake can lead to
severe consequences, including reduced feed intake and potential starvation (Ouyang et al.
2021). MPs have been also shown to interfere with digestive enzymes, trigger the release
of immune cytokine digestive tract in guppy fish (Huang et al. 2020), cause behavioral
changes, affect energy reserves and food composition, and disrupt liver function in guppy
(Lai et al. 2021; Yin et al. 2018). In addition, MP infiltration into fish muscle tissue is also
important from a consumer health perspective (Yang et al. 2020; Zitouni et al. 2020) as
MPs may impact human health (Hossain et al. 2023b).
As MP pollution in aquaculture has garnered increasing attention in recent years, BFT
may emerge as a promising mitigation strategy (Hossain et al. 2023b; Meng et al. 2023),
especially due to its rich microbial biodiversity (Hu et al. 2023). The exponential growth
of diverse microbial population including fungi, protozoa, and bacteria (Khanjani et al.
2022) may enable a fast MP decomposition (Abakari et al. 2022). These microorganisms
can break down the plastic into smaller molecules, which can then be further degraded by
other processes (Li et al. 2022; Wu et al. 2022). The study found that the bioflocculants
produced by Bacillus enclensis had a flocculating activity of 93%, and they were able to
reduce microplastic pollution by up to 80% (Shukri et al. 2022). Amini et al. (2022) found
that a variety of bacteria, including Pseudomonas sp., Bacillus sp., and Vibrio sp., have
been shown to be able to degrade microplastics. In addition, biodegradation of microplas-
tics was observed in Pseudomonas sp. MB4 at a rate of 20% per day (Huang et al. 2022a),
Bacillus cereus B-1 at a rate of 10% per day (Li et al. 2022), and Pseudomonas aeruginosa
B-3 at a rate of 15% per day (Wu et al. 2022), both groups commonly present in BFT aqua-
culture conditions. Those rates can be affected by numerous factors, such as the type of
plastic, the concentration of bacteria/fungi, and the environmental conditions (Amini et al.
2022; Meng et al. 2023).
In BFT systems, Hu et al. (2023) examined the impact of polystyrene particles with
32–40 µm diameter range in Nile tilapia (O. niloticus) cultivation for 28 days. The study
revealed that exposure to 0.80 and 800 particles/L did not affect water quality (including

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nitrification), water (biofloc) and gut microbial communities, tilapia growth, or digestive
enzyme activity. However, changes in crude protein and lipid content were observed, as
well as an increase in the hepato-somatic index, suggesting potential interference with liver
digestive absorption. The authors revealed that MPs were not detectable in tilapia feces
from day 16. The highest concentration of MPs was found in the liver followed by the gills,
while microparticles were not observed in muscle tissue. These studies may indicate that
BFT microorganisms may tolerate water containing MPs without affecting water quality,
animal performance, nitrification, and other key microbial properties. It may also help to
degrade MPs, thereby presenting a promising area for future research into MP remediation.
However, due to its limitation, further toxicological studies are needed to better illustrate
the role of BFT aiming to overcome MPs challenges in aquaculture (Wei et al. 2021, 2022).

Conclusions and perspectives

The present review emphasizes the importance of proper water quality management in
BFT, and to maintain key parameters such as N-compounds, temperature, salinity, TSS,
SS, pH, DO, and alkalinity in optimal levels. Routine monitoring, interpretation, and inter-
vention are essential to sustain a desirable environment, animal welfare, health and perfor-
mance. There is still a lack of understanding related to water quality and BFT, especially
in commercial biofloc-based shrimp and fish operations. Developing a comprehensive bib-
liometric analysis, this review intended to combine relevant water quality literature in fish
and shrimp biofloc systems, and support decision-making to improve farming operations.
In the future, BFT is expected to play a major role as suitable technology for pond water
quality management. The present study underscores the importance of understanding the
complex interactions within BFT systems and the potential of bioflocs to mitigate the envi-
ronmental impacts, including HMs and MPs.

Author contributions Mohammad Hossein Khanjani, Alireza Mohammadi, Maurício Gustavo Coelho Eme-
renciano wrote the main manuscript text.
Mohammad Hossein Khanjani, Alireza Mohammadi, Maurício Gustavo Coelho Emerenciano prepared
figures and tables.
Mohammad Hossein Khanjani, Alireza Mohammadi, Maurício Gustavo Coelho Emerenciano reviewed
the manuscript.

Data availability No datasets were generated or analysed during the current study.

Declarations
Competing interests The authors declare no competing interests.

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Authors and Affiliations

Mohammad Hossein Khanjani1 · Alireza Mohammadi2 ·

Maurício Gustavo Coelho Emerenciano3

* Mohammad Hossein Khanjani

[email protected]; [email protected]
1
Department of Fisheries Sciences and Engineering, Faculty of Natural Resources, University
of Jiroft, Jiroft, Kerman, Iran
2
Department of Environmental Sciences and Engineering, Faculty of Natural Resources, University
of Jiroft, Jiroft, Kerman, Iran
3
Commonwealth Scientific and Industrial Research Organization (CSIRO), CSIRO Agriculture
and Food, Livestock & Aquaculture Program (Aquaculture Systems Team), CSIRO Bribie Island
Research Centre, Woorim, Australia

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