































Highlights in BioScience
ISSN:2682-4043
DOI:10.36462/H.BioSci.202201

Review Article

Open Access

1 Biofuel Research Laboratory, Department of

Microbiology, School of Life Sciences, Central

University of Tamil Nadu, Thiruvarur, Tamil

Nadu, India.

* To whom correspondence should be
addressed: suchitrar@cutn.ac.in

Editor: Hatem Zayed, College of Health and
Sciences, Qatar University, Doha, Qatar.

Reviewer(s):
Alsamman M. Alsamman, African Genome Center,
Mohammed VI Polytechnic University,Morocco..

Morad M. Mokhtar, Agricultural Genetic
Engineering Research Institute, Agricultural
Research Center, Giza, Egypt.

Received: October 29, 2021

Accepted: January 1, 2022

Published: January 15, 2022

Citation: Ray B, Rakesh S . Phycoremediation of
aquaculture wastewater and algal lipid extraction
for fuel conversion. 2022 Jan 15;5:bs202201

Copyright: © 2022 Ray and Rakesh. This is
an open access article distributed under the terms
of the Creative Commons Attribution License,
which permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited.

Data Availability Statement: All relevant data are
within the paper and supplementary materials.

Funding: This work is financially supported
by SERB DST project (EEQ/2018/001463).

Competing interests: The authors declare
that they have no competing interests.

Phycoremediation of aquaculture wastewater and algal lipid extrac-
tion for fuel conversion

Bobita Ray1
>< ,Suchitra Rakesh*1

>< 

Abstract
In this review, it is discussed the prominent effect generated from aquaculture wastew-

ater considered as the major water polluting crisis in the entire world. The cause rose due
to intense development and improvement in aquaculture by the aquatic habitat species trig-
gering quite a challenge in the environment. Scrutinizing this problem, researchers have
found a way to tackle it by cultivating algal species in aquaculture wastewater in order to
remove its high content of organic and inorganic pollutants. The theory proves wastew-
ater serves as a nutrient source for algal growth and development such as phosphorous,
nitrogen, and other trace elements. Besides harvesting the algal biomass from aquaculture
wastewater, the extraction of lipid is also processed for biofuel production. Hence, the dis-
cussion includes conversion of wastewater into organic and inorganic pollutant-free water
with low cost-effective method via algal cultivation in wastewater and high lipid yield for
biofuel with a carbon-free and sustainable environment.

Keywords: Algae, aquaculture wastewater, harvesting, lipid extraction, transesterification

Introduction
Ever increasing global population and continuous dependence of fossil fuels, increased urban-

ization and industrialization posing a major threat to energy security and environmental concerns

to both developed and developing nations. With the accelerated speed of increasing population

growth, wastewater treatment is considered as one of the solutions to control the environmental

issues. And the additional challenges for water scarcity bring out the crucial problem related to

wastewater. Hence, much of the emphasis has been given now a days for wastewater treatment [1].

The anthropological activities such as sewage, industries, agriculture, medical, research laboratories

etc., are pointed to be the sources of wastewater which have tremendously polluted the water re-

sources. Wastewater from various sources comprises both organic and inorganic pollutants. Organic

pollutants include proteins, carbohydrates, lipids, etc., whereas inorganic mostly have chemicals

and solvents [2], and in industrial wastewater even heavy metals or toxic elements are reported

[1]. Nowadays the aquaculture wastewater is quite prominent globally due to its intense develop-

ment and improvement in fish, marine species, algae and aquatic plant farming. Thus, this rapid

increase of aquaculture effluents poses a serious threat to environment [3]. Mariculture other name

of aquaculture can be seen their progresses in parallel way with high human demands. The impact

performances to the environment for food production has been reported as the fastest growth [4].

Aquaculture wastewater contains a high number of pollutants and chemicals that lead the residing

aquatic flora and fauna to die due to eutrophication. The toxic algal blooms not only disturb the

aquatic life but it also interferes with the sustainable marine aquaculture development. The wastew-

ater treatment via biochemical methods is not economical and further removal of those chemicals

from water poses another challenge. Hence, wastewater treatment by algae is an ecofriendly and

cost-effective approach over other physical and chemical methods [5–7].

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https://orcid.org/0000-0002-4357-4042
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Ray and Rakesh, 2022 Phycoremediation of aquaculture wastewater for fuel conversion

Earlier aquaculture wastewaters were treated with bulks of
antibiotics which later have evolved to antibiotic-resistant [8].
But in an investigation, green microalgae Tetraselmis sp. re-
moved nitrogenous and phosphorous compound from aquatic
wastewater within 48 hours [9]. Removal of nutrients from live-
stock wastewater was also reported via, Desmodesmus sp. a
microalga with potential benefactor [10]. The wastewater treat-
ment via microalgae not only removes the pollutants but also
shows positive effect towards carbon fixation (1.83 kg CO2/kg
of biomass), high amount of biomass generation within a short
period of time. The microalgal biomass can be further utilized
for biofuel and valuable bio-products production and can also
act as substrate in bio-refinery. Thereby, it provides a sustain-
able and ecofriendly approach to many of the problems related
with wastewater [11].

Since the middle time of 20th century, development in aqua-
culture growth has risen globally in all over countries providing
huge profit to commercial hatcheries and farming system. The
wastewater discharge from chemical and other industries has
high toxicity level, that poses serious environmental issues [12].
The aquaculture production is kept on increasing due to high de-
mand in the market. Hence large-scale production has been car-
rying out enormously [13]. Yang et al., [14] has mentioned in
his studies the aquaculture effluent treatment with microalgae is
highly efficient in absorbing nutrients and value-added biomass
generation.

Alga-aquaculture has led to many advantages such as com-
pared to other plants, algae has proved to be better in nutrient
removal. The construction and operational costs are low with
consistent to high nutrient removal efficiency. The microorgan-
isms consortia like algal-bacterial consortia are highly efficient
in solid and other waste treatment into low molecular weight
compounds [15]. The large quantity of algal biomass can be
produced from aquaculture, that can be further utilized for high
demand valuable product generation Spirulina and Chlorella cul-
tures are commonly used as aquaculture feed, as both has very
minimal toxicity level and helps in preventing algal blooms as
well. On the other way, addition of expensive chemicals and
antibiotics for industrial effluent treatment are not economical
and poses severe threat to the environmental [16]. It has also
been reported that excess use of chemicals in aquaculture af-
fects the food safety and quality of meat produced via aqua-
culture. In most of the research studies, microalgae such as
Chlorella sp. is found experimenting in every aspect of wastew-
ater treatment. Biofilms are another slimy and foul in nature
found on the surface of the algae or any solid surface attached.
Microalgal biofilms mostly have succeeded in reducing the nu-
trients of phosphorous starting initially from 15 mg L-1 within
24 hours[12]. The biofilm has succeeded in more production of
biomass production for biofuel [17].

This review summarizes our efforts towards various aquacul-
ture wastewater treatment via microalgae and use of algae for
feed purposes. It further highlights the biofuel and value-added

products generation from the algal biomass.

Micro and macro algae as a nutritive aquaculture feed
Aquaculture has been rapidly developing in industrial sector

resulting large quantity of polluted effluents being discarded into
clear water line. Remaining solid residues mostly contains haz-
ardous chemicals and metal elements causing severe incurable
diseases [1]. Huge amount of cost and labor are invested upon
various physical and chemical techniques. Electro adsorption
and electro-reaction coupling process is one of physical tech-
nique to clean the wastewater, removing 99% of solid. But it is
reported algae is the most efficient and advanced method with
low cost benefits in treating wastewater [18]. In aquaculture,
many of the microalgae viz., Nannochloropsis, Chaetoceros, Tha-
lassiosira, Tetraselmis etc. are known for essential food sources
including marine species such as clams, molluscs, oysters and
Spirulina sp. for providing high protein diet for freshwater fishes
and other invertebrate species [17]. Treatment of wastewater
with microalgae has been guaranteeing good outcome and have
led to great advantages without harming the environment.

Macroalgae universally known as seaweed is easily visible
through naked eye. Its habitats are mostly native to marine or
other river bodies. Saccharina latissima also known as kelps are
generally found in river depth. Macroalgae are well-known for
their nutritional and bioactive components. Aquaculture with
macroalgae production has a strong demand in the market, ac-
cording to industrial vision. It is a valuable source of aquaculture
feed. However, most examples of eutrophication in the marine
environment are caused by the deposition of flowing waste in
the sea, which includes high nitrogen and phosphorus nutrients.
Macroalgae also aids in the bioremediation process by prevent-
ing wastes from impacting the marine environment in terms of
pH change, turbidity, and increased BOD content, as well as
causing marine life death and encouraging toxic algal blooms
[19–21]. According to Brakel et al., [22] macroalgae depicts as
fastest growing aquaculture development even in poorest coastal
regions. Nowadays with advanced facilitation and support of ge-
netic resources, seaweeds such as red algal genera Eucheuma
and Kappaphycus proved economically in many tropical coun-
tries.

Microalgae based biorefinery for aquaculture wastew-

ater treatment
Wastewater treatment is rising as fundamental priority. The

removal of nutrients and solids, as well as the acceptance of envi-
ronmentally friendly remediation techniques, play a significant
role in this. The most photosynthetic machinery technique that
we can ever expect is phycoremediation, or treating wastewater
with algae. It extracts all unwanted parameters from wastewa-
ter and improves water quality to meet environmental standards.
[15]. Compared to conventional wastewater treatment this method
is cheap and also has the involvement in biomass production for
biorefinery purposes. Chlorella sorokiniana is observed as the

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Ray and Rakesh, 2022 Phycoremediation of aquaculture wastewater for fuel conversion

most utilized objective both for phycoremediation and biomass
production [23].

Open pond system
Algal cultivation is the basic necessary condition for more

quantity of biomass for biofuel production. Depending on cost,
labor and time vast kinds of techniques are available. Open pond
is the common system for large scale algal production exposed
directly to environment. Here generally, algae species are cul-
tivated in an open pond area covering as much acres of land
directly under the sunlight due to their phototrophic nature [24].
Open pond is named as raceway because it resembles with race-
track. This raceway pond system takes less space of land for
growth. It requires continuous movement of paddle wheel in
the pond to prevent sedimentation of cultures at the bottom level
[25]. Generally, paddle wheels depicts the main principle base
for the open pond system where the speed of the wheel helps to
cover the light intensity for all over the algal growth within the
system [26].

Nutrient removal efficiency of aquaculture wastewater

with microalgae
The most efficient process and cost-effective method for cul-

turing algae is via wastewater sources; rather than cultivating
in expensive amounts of chemicals. Aquaculture wastewater
contains required nutrients such as nitrogen, sulphur, phospho-
rous which alga feeds on for growth. Its composition is men-
tioned in Table 1 , pointing its physico-chemical properties such
as its pH, Chemical Oxygen Demand (COD), nitrate, chloride,
sodium, potassium, magnesium, nitrite, ammonium and phos-
phorous were depicted in aquaculture wastewater. In Table 1,
the content found under those properties extremely higher com-
pared to normal i.e. these wastewater has the capability to cause
diseases.

Wastewater cultivation is positively progressing both in biore-
mediation and biomass production for biofuel. It is either way
sustainable to environment as budget friendly way. Ulva sp.,
Codium sp., Ecklonia sp., Saccharina sp., Gracilariopsis sp. have
experimented in fish seaweed aquaculture waste for bioremedia-
tion that have removed high concentration of ammonia and phos-
phorous within 30-40% nutrient removal converting into less
polluting [30]. Whereas for microalgae Tetradesmus obliquus
has removed 99.3% of ammonia and 99.2% of phosphorous con-
centration from swine manure wastewater [31]. In Table 2, var-
ious algal species cultivated in different types of aquaculture
wastewater are shown. The inoculated culture in the wastewater
is mentioned parallel to the algal species name. The remaining
columns are about the results of removal of nutrients described
in percentage that found after cultivating in aquaculture wastew-
ater. It specifies how algae worked as bio-remediation.

Ta
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.

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as
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er
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O

D

(m
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L
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N
itr

at
e

(m
g/

L
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C
hl

or
id

e

(m
g/

L
)

So
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um

(m
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ta

ss
iu

m

(m
g/

L
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ag

ne
si

um

(m
g/

L
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N
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ite

(m
g/

L
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A
m

m
on

iu
m

(m
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)

Ph
os

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or

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(m
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7.
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Highlights in BioScience Page 3 of 9 January 2022|Volume 5

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Ray and Rakesh, 2022 Phycoremediation of aquaculture wastewater for fuel conversion

Table 2. Nutrient removal in aquaculture wastewater with different algal species

Types of aquaculture wastewater Algae species Type of algae
Amount of

inoculation

(g/L)

Time of

treatment

(Days)

Removal compounds
Reference

COD

(%)

NH4

(%)

Nitrate

(%)

Nitrite

(%)

Phosphorous

(%)

Total nitrogen

(%)

Fishery P. kessleri TY Microalgae 10 3 94.4 96.2 94.3 99 96.6 NA [7]

Salmon farming Chlorella minutissima Microalgae NA 10 NA NA 88.6 74.3 99 88 [27]

Shrimp culture Gracilaria tenuifrons Macroalgae 1.75 8 NA 35.1 NA 71.7 33.2 2.8 [28]

Fish-seaweed aquaculture Codium fragile Macroalgae 1000 28 NA 0.07 NA NA 0.22 0.56 [23]

Ulva pertusa 1000 28 NA 0.04 NA NA 0.15 0.56

Ecklonia stolonifera 1000 28 NA 0.11 NA NA 0.26 0.57

Gracilariopsis chorda 1000 28 NA 0.11 NA NA 0.23 0.50

Saccharina japonica 1000 28 NA 0.15 NA NA 0.21 0.56

Oreochromis niloticus aquaculture Chlorella sorokiniana Microalgae NA 14 NA 99.9 75.2 NA 77 78 [29]

NA Not Available

Recent advances in microalgae harvesting and lipid ex-

traction
After cultivation, harvesting which means collecting or gath-

ering of algal cultivation determines as most difficult and im-
portant out of all process work. For large scale harvesting of
biomass, it requires quite expensive technique, maintenance of
time, man power and so on. In case of microalgae harvest-
ing techniques such as centrifugation, sieving, filtration, sedi-
mentation, flotation, flocculation are predominantly utilized [33].
whereas for macroalgae simple technique such as drying and
storing is basically preferred but however few techniques from
microalgae harvesting techniques are also operated [34]. Thermo
reversible gel transition [35,36] characterized with either agar or
sol gel for harvesting of algae where clustered cells are settled
at bottom and collected the biomass. Flocculation is another
technique of harvesting. Nanocellulose is an insoluble substance
where bonding of polysaccharide and glucose monomers occurs
with the concept of more concentration of nanofibril more in-
crease of flocculation [37]. Bacterial cellulose Gluconacetobac-
ter xylinus has found to be successfully harvested with 90% of
clump formation [38]. Pleaurotus ostreatus [39] and Scenedesmus
obliquus [40] are another flocculating process. According to
Leite [41], pH modulation through Dissolved Air Flotation can
be harvested at higher biomass. Magnetic nanoparticles is an-
other better technique for harvesting [42]

Mostly utilized lipid extraction method is Bligh and Dyer
as said to be quickly approachable to quantification outcome
within less timing but more hazardous to environment as well
as self-health. But MTBE i.e., Methyl-tert-butyl ether is the bet-
ter method than the previous method with non-hazardous effect
and increase in the extracted lipid [43]. For future perspective
role such as to study the characterization from extracted algal
biomass production, high resolution nuclear magnetic resonance
spectroscopy (HR NMR) or mass spectroscopy technique is used
to study the changes of various composition kept in different
storage conditions were found in the algal sample. Such tech-

niques are extremely advanced in analytical process [34]. On
the other hand, Dimethyl ether is a gas type where the liquefied
gas is passed with the help of Nitrogen gas and proceeded for al-
gal extracting [44]. Super high hydrostatic pressure technique is
even utilized for extracting lipid maintaining pressure 100 MPa
to 1000 MPa [45]. Another process of lipid extraction solvent
is ionic liquid that comprises of ion solvents of non-volatile sub-
stance including bubbling CO2 gas for extraction [46]. In the
given Table 3, it basically determines the biomass productiv-
ity and lipid growth found after algal cultivation in aquaculture
wastewater. In the same table various algal species names and
according to that in the left column the types of wastewater are
given where following that horizontally we can read the biomass
and lipid found after cultivating the algae in that same wastew-
ater. Within these three table tables (Table 1, 2 and 3) it gives
the idea about reading the physico chemical properties before al-
gae cultivating and harvesting the biomass, lipid measuring and
lastly with the remaining water and can be proceeded with re-
reading the physico chemical properties determining removal of
nutrients from the wastewater.

Microalgae as a sustainable future biofuel approach
Developing with rapid high rise of industries by regular use

of natural resources are leading us into depletion of fossil fuels
sooner creating havoc in environment. It surges carbon dioxide
till peak point making possibly prone to global warming simi-
larly threatening wastewater globally. At the bright side, the mi-
croalgae have several unique features like ability to fix CO2 and
convert it into valuable components via photosynthesis, robust
growth with high lipid contents. The microalgae harvesting,
qualitative and quantitative estimation of lipid has been reviewed
[43,47]. The availability of molecular approaches to increase
lipid accumulation and recovery has been extensively discussed
[48]. Biofuel is the breakthrough for solution. Among gener-
ation after generation there has been change into biofuel pro-
duction. Initially beginning with edible plants such as soybean,
maize, brassica comes under first generation and had a great deal
with alternative fuel. The lipid yield was good but in case of

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Ray and Rakesh, 2022 Phycoremediation of aquaculture wastewater for fuel conversion

Ta
bl

e
3.

N
ut

ri
en

tr
em

ov
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in
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ua
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ltu
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w
as

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at
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w
ith

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ff

er
en

ta
lg

al
sp

ec
ie

s

A
qu

ac
ul

tu
re

w
as

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w

at
er

A
lg

al
sp

ec
ie

s
Ty

pe
of

al
ga

e

To
ta

ld
ur

at
io

n
da

ys

of
tr

ea
tm

en
tm

et
ho

d

(m
g/

L
)

B
io

m
as

s
co

nc
en

tr
at

io
n

/p
ro

du
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iv
ity

(%
)

L
ip

id
gr

ow
th

(%
)

R
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Fi
sh

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P.
ke

ss
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TY

M
ic

ro
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5
26

N
A

[7
]

Se
a

br
ea

m
fa

ct
or

y
Te

tr
as

el
m

is
su

ec
ic

a
M

ic
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10

68
25

[2
7]

Sa
lm

on
fa

rm
in

g
C

hl
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la

m
in

ut
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si
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a
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ic
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10

55
46

.3
7

[1
3]

N
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e

other matters like production of biodiesel from food crops dur-
ing the time of world war period was a huge downfall. In second
generation Jatropha plant being the non-edible is another alter-
native fuel production which is a good source compared to first
one [49]. The life cycle, production in large scale, huge mass of
land for cultivation is the major demerit. Third generation i.e.,
microalgae is currently the most successfully running lipid yield
production out of all. Cultivation of microalgae is only 14 days
where this microorganism can be grown in even a small tub or ar-
tificial huge ponds. The biomass with high production of yield
can be grown in any suitable environment with different stress
conditions changing physiological condition. Monoraphidium
sp.is cultivated in BG 11 media and transferred into high am-
monia content wastewater with 44% of stress condition present
[50].

Transesterification, in case of biodiesel it can be termed as
conversion of a 3-methyl glyceride when it reacts with methanol
in presence of catalyst to form Fatty acid methyl esters to form
ethanol, likewise shown in Figure 1. For conversion into biofuel
after harvesting method and weighing dry biomass, lipid extrac-
tion process is followed. It basically consists of two types me-
chanical and non-mechanical, the previous type usually relates
with solvents and the later describes extraction through instru-
mental techniques. Triglycerides act as main components, these
are fatty acids extracted from algal species and converted into
fatty acid methyl esters through direct transesterification method.
This method depicts reaction of triglycerides with mono alco-
hols in presence of catalyst were analyzing solvent as hexane
with better results compared to chloroform and methanol where
pointing a strong line selection of solvents affects in lipid yield-
ing after purification [51].

Biodiesel has inherent sustainable transportation fuels for fu-
ture mostly to reduce increasing pollutants emitted from exhaust
cylinder. Many modernized machine learning techniques and re-
newable feedstock are emerging rapidly for biofuel conversion
compared to chemical catalysts. The main source of biodiesel
is manufactured basically from renewable oil derived microbes
or plants which causes zero-effect in ecosystem accompanying
with carbon reduction. Enzyme mediated undertakes non-toxic
transesterification compared to same old process of chemical uti-
lization [52,53].

From many processing experiments, microbes such as mi-
croalgae is the leading aspect, Euglena sanguinea due to its pres-
ence of superior combustion characteristics were able to produce
biodiesel that blends with the regular agricultural diesel engine
till 40% by extracting lipid from the algal biomass [27]. A het-
erogeneous nano-catalyst Ca(OCH3)2, a novel reactive distilla-
tion column is experimented for algal biodiesel production opti-
mized by maximizing biodiesel purity by NSGA-II, non-dominated
sorting genetic algorithm, designed both for low cost produc-
tion and CO2 emissions [28]. For another substitute yield of
biodiesel an experiment conducted between Chlorella sorokini-
ana and Monoraphidium sp. where the biomass, fatty acid pro-

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Ray and Rakesh, 2022 Phycoremediation of aquaculture wastewater for fuel conversion

Figure 1. Flowchart diagram for biodiesel production through use of transesterification.

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Ray and Rakesh, 2022 Phycoremediation of aquaculture wastewater for fuel conversion

file studies were compared showing better outcome from Chlorella
sp. In the mentioned study, lipid analyzing, its thermal effi-
ciency all were recovered higher for biodiesel benefitting with
low emissions of CO and HC [51]. Lipid extraction through
microwave assisted in situ transesterification technique for the
biodiesel yield was achieved from algae such as Ulothrix sp.
88% dry weight (DW), Cladophora sp. 80% DW, Oedogonium
sp. 73% DW and Spirogyra sp. 67% DW. This technique for
high biodiesel yield utilizes solvent free method [29].

Conclusion
From this study it reveals aquaculture wastewater is being

the significant source for cultivating algae proceeding with sus-
tainable environment in simple, cost effective way with zero
waste reassurance. Both microalgae and macroalgae plays vi-
tal role in aquaculture production and wastewater remediation.
The presence of nutrients in wastewater reveals necessary re-
quirements for their growth. The emitted aquaculture effluent
contains highly nutritive source for algae that blends into it rec-
ommending as bio or phyco remediation in process. Addition to
that biomass produced from aquaculture wastewater can also be
converted into biofuel through transesterification process with
recent ideas of harvesting techniques. From the reported articles,
it is known not much work have been proceeded in aquaculture
wastewater co-related with micro and macroalgae. It still needs
to be explored in order to achieve higher biomass and lipid for
biofuel production where a solution is required for further re-
search as there is huge gap in laboratory work and large-scale
production. Hence, this study needs to be taken to further sim-
plified step by investigating more into it.

Acknowledgement
The authors thank the academic writing group and SWAYAM

MOOCs course initiated by the Ministry of Human Resource De-
velopment, Government of India, for providing an open learning
platform.

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	Abstract
	Introduction
	Micro and macro algae as a nutritive aquaculture feed
	Microalgae based biorefinery for aquaculture wastewater treatment
	Open pond system
	Nutrient removal efficiency of aquaculture wastewater with microalgae
	Recent advances in microalgae harvesting and lipid extraction
	Microalgae as a sustainable future biofuel approach
	Conclusion
	Acknowledgement
	References

