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Contents lists available at ScienceDirect

Process Safety and Environmental Protection

journal homepage: www.elsevier.com/locate/psep

Direct brackish water desalination using Chlorella vulgaris microalgae

Malihe Barahoei a , Mohammad Sadegh Hatamipour a,∗ , Saeed Afsharzadeh b
a
Department of Chemical Engineering, Faculty of Engineering, University of Isfahan, Isfahan, Iran
b
Department of Plant and Animal Biology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran

a r t i c l e i n f o a b s t r a c t

Article history: The objective of this study was to investigate the suitability of direct use of Chlorella vulgaris for brackish
Received 25 June 2020 water desalination as a new conceptual technique. First, the adaptation of Chlorella vulgaris in saline water
Received in revised form 5 October 2020 was performed, and then the living microalgae cells were utilized for the desalination process using a
Accepted 6 October 2020
bubble column photobioreactor. The effect of culture medium, time, salinity and initial inoculum on the
Available online xxx
microalgae growth and salinity removal was investigated and the optimum conditions were obtained by
RSM−CCD method. To assure the consumption of sodium chloride (NaCl) content of water by microal-
Keywords:
gae, the BG11 culture medium was modified by substituting its chloride and sodium containing salts
Desalination
Microalgae
by nitrate, calcium and potassium containing minerals. The results indicated that the enhancement of
Bubble column photobioreactor microalgae growth and salt removal efficiency were more pronounced in the modified-BG11 (MBG11)
Chlorella vulgaris culture medium. Using Chlorella vulgaris microalgae in the MBG11 culture medium, the decrease in brack-
BG11 culture medium ish water electrical conductivity for different NaCl concentrations between 1000 and 5000 ppm, was
between 80 % and 40 %, respectively. Atomic absorption and flame photometry analyses confirm the
hypothesis of adsorption of Na+ ions on the Chlorella vulgaris cell surface.
© 2020 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.

1. Introduction The desalination technologies broadly used around the world

Due to the worldwide population growth and the reduction of
freshwater resources, the scarcity of fresh water has become a very
hazardous issue. More than one billion people do not have access to
sanitary drinking water, and nearly 2.3 billion people (about 41 %
of the world’s population) live in scarce water areas (Morillo et al.,
2014).
About 1% of the world’s water is available as freshwater in lakes,
rivers and wetlands. On the other hand, the amount of freshwater
available is unevenly spread around the world (Iaccarino, 2019). In
Iran, due to the extended drought conditions as well as reduction
in annual precipitation during the last years, water supply has been
limited to deep wells, using underground water reservoirs. There-
fore, research development in the desalination field is essential
(Ghalavand et al., 2015).
Desalination processes are used in municipal, industrial or
commercial applications. With the advancement of technology,
desalination processes are becoming the cost-competitive option
for producing usable water.
∗ Corresponding author.
E-mail address: hatami@eng.ui.ac.ir (M.S. Hatamipour).
can be mainly classified as either thermal or membrane techniques.
Both technologies require energy for operation and freshwater pro-
duction. The former includes processes based on phase change,
i.e., distillation or freezing, while the latter includes processes uti-
lizing membranes such as reverse osmosis, forward osmosis or
electrodialysis (Narayan et al., 2012). Overall, Multi-Stage Flash
Distillation (MSF), Multiple Effect Distillation (MED), Vapor Com-
pression (VC), Humidification dehumidification (HDH) and Reverse
Osmosis (RO) are the most common water desalination tech-
nologies. Due to the high production capacity of some of these
systems, their energy consumption is high, and they are usually
built adjacent to power plants to supply this energy. Therefore,
there is a continuous effort to develop water desalination processes
with lower energy consumption (Bohn et al., 2009; Ghalavand
et al., 2015; Sharon and Reddy, 2015; Spiegler and El-Sayed, 2001;
Morillo et al., 2014; Abdelmoez et al., 2014). Biological systems have
the ability of desalination by living microorganisms without any
environmental damage (Taheri et al., 2016; Jlassi et al., 2013; Rabhi
et al., 2010; Ditta, 2016; de Lacerda et al., 2015; Nie et al., 2020).
By developing new biological desalination methods, lower energy
consumption and costs can be achieved primarily due to the less
damaging environmental effects. One of these new biological pro-
cesses for water desalination is the use of microalgae (Taheri et al.,
2016; Kalleary et al., 2014).

https://doi.org/10.1016/j.psep.2020.10.006
0957-5820/© 2020 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.

Please cite this article as: Barahoei, M., et al., Direct brackish water desalination using Chlorella vulgaris microalgae, Process Saf. Environ.
Prot., https://doi.org/10.1016/j.psep.2020.10.006
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Microalgae are a heterogeneous group of organisms which vary Table 1

Chemical composition of BG 11 and MBG11culture media.
in size, having several different shapes. Microalgae can grow in
water, whether it is seawater or freshwater, and under different Chemical (BG-11) BG-11(gL−1 )
climatic conditions ranging from very cold to hot climate. They can NaNO3 1.5
grow wherever they could synthesize their components and feed K2 HPO4 .3H2 O 0.04
by photosynthesis (El Sergany et al., 2014). Microalgae are a wide Na2 CO3 0.02
group of microscopic plants which can contain up to 50–70 % pro- MgSO4 .7H2 O 0.075
CaCl2 .2H2 O 0.036
tein (that is up to 50 % in meat and 15–17 % in wheat), 30 % lipids,
EDTA-Na2 0.001
over 40 % glycerol, up to 8–14 % carotene and vitamins B1, B2, B3, Fe(NH3 )2 Citrate 0.006
B6, B12, E, K, D, etc. (Priyadarshani and Rath, 2012). Citric acid 0.006
Microalgae could absorb carbon dioxide through photosynthe- Trace elements 1 mL
sis reactions and produce biomass and oxygen (Pires et al., 2012; Chemical (MBG-11) MBG-11(gL−1 )
Barahoei et al., 2020a). Therefore, if it is used to eliminate the KNO3 1.78
salinity of water during its growth, freshwater can be obtained K2 HPO4 .3H2 O 0.04
CaCO3 0.009
by a simple desalination technique with the lowest energy con-
MgSO4 .7H2 O 0.075
sumption, whose produced rich biomass can be used in various CaNo3 .2H2 O 0.034
applications (Ho et al., 2011; Kalleary et al., 2014). Microalgae can EDTA-K2 0.001
be used for health, food or their additives, cosmetics, pharmaceu- Fe(NH3 )2 Citrate 0.006
Citric acid 0.006
ticals, biofertilizer and most importantly for production of energy
Trace elements 1 mL
sources such as biofuel and bioethanol.
The adaptation of microalgae to high level of salinity may be
carried out in a process that includes three stages: (1) turgor
restoration, (2) cell membrane adjustment to allow the absorp- with microalgae in the consumption of nutrients and reduces the
tion and export of ions, and (3) induction of the accumulated stress growth of microalgae.
proteins and glycerol as osmoprotective compatible solutes which Another study used Chlorella vulgaris and Scendesmus at differ-
have been produced photosynthetically. Cellular homeostasis with ent salinity concentrations in the range of 2–20 g/L. They cultivated
the influx of NaCl can be degraded, impairing the biopolymers halophile algae (S. sp. and Chlorella vulgaris in photobioreactor
function. Redox-driven sodium pumps can make acclimation of (PBR) to sequester sodium chloride salt from brackish and sea-
microalgae by absorbing and exporting potassium and sodium water. They also investigated the potential of microalgae use for
(Thomas and Apte, 1984). For example, the vacuole of Dunaliella biological desalination. They harvested the valuable lipids from
salina cells can trap the intercellular ions, accumulating protec- microalgae as a byproduct that can be used to generate bio-fuel
tive compatible solutes in its structure (Espie et al., 1988; Bhargava and supply energy back to the desalination process. The doubling
et al., 2003). If potassium ion and organic chemicals (such as glycine time for the microalgae during growth phase was varied between
and proline) can be accumulated in microalgae cytoplasm to bal- 3.1 and 5.9 days for Chlorella vulgaris, and 0.63–1.81 days for Scen-
ance the osmotic pressure, the cell vacuole can sequester the Cl− desmus (Sahle-Demessie et al., 2019). They did not investigate the
and Na+ ions (Fisher et al., 1997). One study found that C. autotroph- consumption of Na+ and Cl− ions as the main salinity factors, they
ica species can accumulate salt in their cells to regulate the osmotic just reported the TDS consumption and biomass production on
pressure (Ahmad and Hellebust, 1984). saline condition. They also didn’t represent the bio-mechanisms
Microalgae (e.g., as Chlorella and Scendesmus) have been widely for the microalgae during TDS removal. Extensive research has not
used for wastewater treatment due to their natural colonization of been done on the salt removal by the direct use of microalgae, and
ponds, fast growth rates and high nutrient uptake capabilities (Nie no in-depth analysis of microalgae activity in salinity removal is
et al., 2020). However, one of their major drawbacks for wastewater available.
treatment is biomass harvesting. Chlorella and Scendesmus are the The development of a new, sustainable, safe and dependable
most active microalgae, because they can survive in a wide range source of freshwater through novel desalination methods seems
of salinity in their habitat (Gimmler et al., 1981). necessary to overcome the water shortage. For this purpose, the
Scendesmus species grows successfully in saline water as it Chlorella vulgaris species were used for the desalination process in a
absorbs the salts, using them in its metabolism. It has been widely bubble column photobioreactor in this work. BG11 culture medium
investigated for desalination (El Sergany et al., 2014; El Nadi et al., was modified to increase its salt removal efficiency and microal-
2014: Sahle-Demessie et al., 2019). Moreover, its superiority in gae growth. Chlorella vulgaris was directly introduced in the saline
operation under suitable laboratory conditions was confirmed (El water to produce freshwater. In addition, an efficient adaptation
Nadi and El Sergany, 2010; El Sergany et al., 2014). A research was method for the microalgae to the salinity can make them resis-
conducted by El Nadi in Egypt to investigate the use of ponds con- tant against high levels of salinity. The adaptation method in this
taining microalgae for water desalination. The microalgae were study was developed for the first time as a novel solution to conduct
introduced into the saline basin at a rate of 0.4 L/basin/run using desalination with microalgae. The response surface methodology
BG11 culture medium. The Salinity concentrations in their study (RSM) based on the central composite design (CCD) was used for
varied from 40 g/L to 80 g/L. The volume of the basin was not men- conducting the experiments and optimization. The effects of salin-
tioned in the article; however, it was estimated to be 420 L based ity, initial microalgae cell number as inoculum for cultivation and
on the analysis of the image presented in the article.Their results time on salt removal efficiency were investigated to identify the
show that the TDS removal efficiency for the mentioned salinity most suitable value of each parameter.
concentrations varied between 13 & 63 %, respectively. These vari-
ations were due to inlet TDS concentration, retention time and
the climatic conditions (Temperature & sunlight period) (El Nadi 2. Material and method
et al., 2014). Since microalgae were prepared in the laboratory and
used to remove salinity from the climatic conditions of the envi- In this study, Chlorella vulgaris and BG11 culture medium were
ronment, it is possible to contaminate the culture medium. On the used. The composition of the BG11 culture medium is shown in
other hand, with the addition of other microorganisms, it competes Table 1.

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The components of trace elements for BG11 culture medium

were Al2 (SO4 )3 .18H2 O, MnCl2 .4H2 O, CuSO4 .5H2 O and ZnSO4 .7H2 O
with the amounts of 3.58, 12.98, 1.38 and 3.2 g L−1 , respectively.

2.1. The habitation of microalgae to different salinities

The Chlorella vulgaris microalgae were adapted to a variety of

salinities at the first stage. To this end, the Erlenmeyer flask was
used as the environment for microalgae culturing. The culture
medium was rotated by a shaker (Vs-203p, vision) at a spin of 120
rpm.
According to our previous work (Barahoei et al., 2020a), different
ranges of light intensity (1700−3700 lux), pH(7–9), temperature
(15−35 ◦ C) and light-dark cycle (12−12,14−10,16−8 hours) have
been tested to determine the values at which Chlorella vulgaris
growth rate was maximized. According to the results, the max-
imum dry weight of Chlorella vulgaris and its growth rate were
obtained when the conditions were adjusted to the optimum
obtained values. Therefore, the used conditions for Chlorella vul-
garis cultivation in this work were a pH of 8, light intensity of 2700
lx, light-dark cycle of 16−8 hours and temperature of 25 ◦ C. In
addition, the one-factor-at-a-time method was used to investigate
the microalgae adaptation behavior at different salinities. In this
method, all variables were fixed at their specified values, except
for only one variable (i.e., NaCl concentration).
The stage of microalgae habitation to salinity was performed
slowly by gradually increasing the NaCl concentration as inchmeal. Fig. 1. Bubble Column photobioreactor for water desalination with algae growth.
This gradual increase in water salinity was necessary for microalgae
adaptation. NaCl was gradually added to the medium in the ranges
2.3. Analytical tests
of 200 ppm–11000 ppm. Upon completion of the growth process
after 14 days, those microalgae cells which survived the NaCl con-
In this study, cell-counting (CL), dry weight (DW) measurement
centration were harvested and entered to a higher salinity of 400
and optical density (OD) spectrophotometry tests were performed
ppm as an inoculator. This process continued up to 11,000 ppm.
to detect microalgae growth. To determine salinity reduction, EC
Therefore, the surviving microalgae cells were harvested after cul-
and TDS were measured.
tivation at 11,000 ppm salinity and used as the salinity reduction
agent.
2.3.1. Cell counting, dry weight and biomass concentration of
microalgae
2.2. Salinity reduction The cell number of microalgal in photobioreactor was mea-
sured using an improved Neubauer hemocytometer, as described
After the habitation of microalgae to high salinity, a bubble col- in our previous study (Barahoei et al., 2020a). The cell number in a
umn photobioreactor was used for their cultivation (Fig. 1). This milliliter sample was calculated by:
photobioreactor was made of a glass cylinder with a diameter of
3 cm and a height of 40 cm. The effective volume of the culture Cells average number = number of cells(per mL of the sample)
medium fluid with microalgae was 250 mL. Light intensity was set
×104×dilution factor (2)
at 2700 lx while the operating temperature was 25 ◦ C. An air flow
rate of 0.5 LPM (liter per minute) was used for aeration in the photo-
bioreactor. By measuring the microalgae optical density over time The optical density of Chlorella vulgaris was measured at 680 nm
using UV–vis spectrophotometer (V-750, Jasco, Japan), the growth wavelength. The pH of the culture medium was adjusted through a
curve for microalgae was obtained. Through monitoring the culture digital pH meter (UB-10, Denver, USA). In addition, the dry weight
medium electrical conductivity (EC) and its total dissolved solids of the microalgae was calculated by :
(TDS) by an electrical conductivity meter (AD 330, Adwa, Hungary),
the salinity reduction was determined using Eq. 1: DW = W2 − W1 (3) (3)

salinity reduction % = 100(EC i − EC f )/EC i (1)
where EC i and EC f are the initial and final EC values of the culture
medium, respectively.
To ensure the consumption of saline water Na+ and Cl− content
by microalgae, the BG11 culture medium was modified by substi-
tuting its chloride and sodium salts with their equivalent nitrate,
calcium and potassium minerals. This culture media is denoted by
MBG11 throughout this article. In this way, NaCl content of the
brackish water could be the only source of Na+ and Cl− intake for
microalgae, which were forced to remove chloride and sodium from
dissolved NaCl.
where W1 is the weight of filter paper and W2 is its weight contain-
ing microalgae filtrate after drying.
2.3.2. Measuring salt uptake and water salinity
One of the necessary parameters for detecting salinity and water
quality in water desalination is electrical conductivity. The electri-
cal conductivity of water is directly proportional to the amount of
salinity. Therefore, an electrical conductivity meter can measure
salinity variations during the cultivation period. Another way to
measure salinity variations is to measure primary and secondary
amounts of Cl− and Na+ ions presented in the brine. In this study,
two methods of atomic absorption and flame photometer were

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Table 2
Ranges and levels of the independent variables by central composite design.
Ranges and levels
Independent variable Symbol
−2
Salinity (ppm) A 0
Time (day) B 3
Initial inoculum (mL/100 mL culture media) C 10
Fig. 2. Microalgae habitation at different salinities.
used to measure primary and final amounts of Na in the culture
medium.
For salinity measurement electrical conductivity meter (AD 330,
Taiwan) and flame- photometer (FP 20, SEAC, Italy) were used to
measure the Na+ content. To ensure the accuracy of the results,
an atomic absorption/flame emission spectrophotometer (AA-670,
Shimadzu, Japan) was also used to measure the Na concentration.
The Na measurement test was conducted on the culture media
and microalgae cells. Therefore, the culture medium in which a cer-
tain amount of NaCl was added to create the desired salinity was
first tested for salinity measurement. Then, adding the inoculation
of Chlorella vulgaris microalgae and after passing 14 days of growth
time, the microalgae solution and culture medium were separated
by a centrifuge (Universal 320, P.I.T, Iran). The separated microal-
gae were washed and dissolved in deionized water. Then it was
sonicated for 30 min with an ultrasound probe (HD2200, Bande-
line, Germany). The resulting sample was used to measure the Na
content by flame photometry.
2.4. Experimental design and optimization by response surface
methodology
Culture medium EC was selected as the response and the opti-
mization was performed using the statistical approach. In the
present study, 40 experimental runs were generated by Design-
Expert software 7.0.0 (Stat-Ease Inc., Minneapolis), including 20
runs for each categoric factor.
Three numeric factors were considered having 5 levels, while
one categoric factor was studied in 2 levels, with six central points
for each level of the categoric factor. The variables together with
their levels are tabulated in Table 2. The experimental results were
fitted with a power equation by multiple regression analysis.
3. Results and discussion
3.1. Adaptation of microalgae in high salinities
The habitation of microalgae at different salinities was per-
formed at the first stage of experiments by one-factor-at-a-time
method. This step was carried out to gain maximum salinity reduc-
tion. Fig. 2 shows the habitation of microalgae at salinities in the
range of 0 ppm–11000 ppm. The color of the medium was changed
by salinity variation from green to nearly yellow.
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−1 0 +1 +2
1000 2000 3000 4000
6 9 12 15
15 20 25 30
In the Erlenmeyer flasks with higher salinity, the color of the
microalgae medium was less intense. When increasing salinity,
the changes in color of samples from green to yellow may be the
result of decreasing the chlorophyll content of Chlorella vulgaris at
higher salinities. The low chlorophyll indicated that the salt stress
and nutrient deficiency limited the microalgae to spend excessive
energy on the synthesis of lots of new chlorophyll molecules and
binding proteins (Sahle-Demessie et al., 2019).
The green color in high salinities (up to 11,000 ppm) indicated
that several microalgae cells remained alive at such high levels
of salinity and were able to grow; they were habituated. Despite
Chlorella vulgaris species salt tolerance and adequate growth, their
significant alteration in salt uptake could be the result of their
salinity habitation mechanism. If the vacuole of Chlorella vulgaris
cell accumulated the Na+ and Cl− ions, enzyme activities could be
inhibited within the cytoplasm (Sahle-Demessie et al., 2019).
During the adaptation stage, the microalgae underwent a multi-
stage process which included the readjustment of osmotic and ionic
potentials as well as wider physiological changes, including regu-
lation of turgor in response to salt stress (Blumwald et al., 1983).
Using photosynthetic bacteria, Amezaga et al. (2014) introduced a
mechanism for bio-desalination. The ATP as the carrier of chemical
energy, powers Na+ export from cells directly or indirectly which
means by Na+ -pumping ATPases or H+ -ATPases (Amezaga et al.,
2014).
Environmental manipulations after the growth phase would
halt Na+ export, and light energy is used to absorb Cl− by
halorhodopsin. The existence of Cl− in the cell draws Na+ through
permeable channel proteins (Amezaga et al., 2014).
Fig. 3 shows the cell counting and microalgae concentration at
different salinities. According to the Figure, the microalgae growth
rate decreased when NaCl concentration increased. On the one
hand, as salinity increased, microalgae growth decreased; never-
theless, the Chlorella vulgaris cells were still alive.
At high salinity (about 11,000 ppm), the surviving cells were
accustomed to such high salt concentrations. They were able to
grow and reproduce. These saline-accustomed cells were isolated
and used for desalination.
In the adaptation stage, when the salt concentration increased
beyond 4000 ppm, a sharp decrease in microalgae growth and cell
number was observed. It was due to the dissolved NaCl in the form
of Na+ and Cl− ions which disrupted the balance of K+ /Na+ in the
algal cells.
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Table 3
The culture media EC results designed by design expert.

Run Salt concentration (ppm) Time (day) initial inoculum (mL/100 mL of culture media) culture media EC (ms)

1 2000 9 20 Modified-BG11 2.02

2 2000 9 20 BG11 4.98
3 1000 6 15 BG11 3.24
4 3000 12 25 Modified-BG11 2.8
5 3000 6 15 Modified-BG11 5
6 1000 6 25 BG11 2.98
7 1000 12 15 BG11 2.38
8 3000 6 25 Modified-BG11 4.2
9 2000 9 20 BG11 5.1
10 2000 9 20 Modified-BG11 2.18
11 1000 6 15 Modified-BG11 2.9
12 3000 6 25 BG11 7.42
13 2000 9 20 Modified-BG11 2.26
14 2000 9 20 Modified-BG11 2.32
15 4000 9 20 BG11 9.58
16 4000 9 20 Modified-BG11 6.46
17 2000 9 20 Modified-BG11 2.26
18 2000 9 30 BG11 4.98
19 3000 12 15 Modified-BG11 2.8
20 2000 9 20 Modified-BG11 2.46
21 0 9 20 Modified-BG11 1.14
22 2000 9 10 BG11 5.26
23 2000 3 20 BG11 5.78
24 2000 9 30 Modified-BG11 1.94
25 1000 6 25 Modified-BG11 1.94
26 3000 6 15 BG11 7.56
27 2000 9 10 Modified-BG11 3.24
28 2000 9 20 BG11 5.02
29 2000 9 20 BG11 5.12
30 2000 15 20 Modified-BG11 1.8
31 1000 12 15 Modified-BG11 1
32 1000 12 25 Modified-BG11 0.76
33 1000 12 25 BG11 2.24
34 3000 12 15 BG11 7.22
35 0 9 20 BG11 1.18
36 2000 9 20 BG11 4.78
37 3000 12 25 BG11 7.2
38 2000 3 20 Modified-BG11 5
39 2000 9 20 BG11 4
40 2000 15 20 BG11 4.82

such as proline, glycine betaine, sugars, polyols and amino acids.

Fig. 3. The cell counting of Chlorella vulgaris at different salinities in different days.
One justification for these phenomena is the fact that unicellu-
lar eukaryotic microalgae often show metabolic activities and share
similarities with higher plants. However, microalgae differ in their
adaptability to salinity and other stress conditions. Response and
adaptation of cells to different abiotic stresses (i.e. salinity, dehy-
dration, cold, excessive osmotic pressure) occur through different
adaptive mechanisms, such as changes in the morphological and
developmental patterns as well as physiological and biochemical
processes (Hiremath and Mathad, 2010). Adaptation to stress is also
associated with metabolic adjustments that lead to the synthesis
and accumulation of several organic solutes as well as osmolytes
These osmotic adjustments, which are well studied in higher plants
(Hasegawa et al., 2000; Hoque et al., 2007), protect subcellular
structures and reduce oxidative damage caused by free radicals
produced in response to high salinity (Hare et al., 1998; Hong et al.,
1992; Hiremath and Mathad, 2010).
By increasing the concentration of the Na+ and Cl− ions above
the osmotic pressure, the operation of the pump was disrupted.
Under such conditions, the microalgae could not properly perform
the growth and propagation process (Barrero-Gil et al., 2005; Falhof
et al., 2016; Eyster, 1958; Arora et al., 2019). At higher salinities,
more microalgae cells were destructed, reducing the number of
cells and the microalgae concentration leading to the lower growth
rate (Zuppini et al., 2010; Arora et al., 2019; Barrero-Gil et al., 2005).
Ultimately, those cells that survived and overcame osmotic pres-
sure at higher salt concentrations were able to grow which were
harvested and used to remove the salt.
3.2. Design of experiments for salinity reduction
3.2.1. Statistical analysis
Analysis of Variance (ANOVA) was conducted for the deter-
mination of the regression coefficients and obtaining a suitable
mathematical model that matches with the variables. Through the
evaluation of F-value, P-value, R2 , R2 Adjusted and R2 Predicted , the
adequacy of the model was determined.
The EC results of the culture media based on the experimen-
tal design are tabulated in Table 3. The ANOVA results, shown in

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Table 4
The ANOVA for Response Surface Reduced Cubic Model.

Source Sum of Squares Df Mean Square F Value p-value Prob > F

Model 29.5556 12 148.402 148.402 < 0.0001 Significant

A-Salinity 15.903 1 958.21 958.21 < 0.0001
B-Time 1.88005 1 113.28 113.28 < 0.0001
C-Initial inoculum 0.21947 1 13.2238 13.2238 0.0011
D-culture media 5.9961 1 361.286 361.286 < 0.0001
AB 0.03744 1 2.25601 2.25601 0.1447
AD 0.84083 1 50.6631 50.6631 < 0.0001
BD 0.61509 1 37.0612 37.0612 < 0.0001
CD 0.09535 1 5.74505 5.74505 0.0237
A2̂ 0.12346 1 7.43862 7.43862 0.0111
B2̂ 0.23716 1 14.29 14.29 0.0008
A2̂D 0.26124 1 15.7404 15.7404 0.0005
B2̂D 0.05759 1 3.47017 3.47017 0.0734
Residual 0.44811 27
Lack of Fit 0.27964 17 0.9764 0.9764 0.5362 not significant
Pure Error 0.16847 10
Cor Total 30.0037 39

Table 5
Statistical parameters for the quadratic model.
Std. Dev. 0.13 R-Squared 0.98
Mean 2.37 R Adj -Squared 0.98
C.V. % 5.44 R Pred -Squared 0.96
PRESS 1.23 Adeq Precision 49.97
Table 4, showed that the presented model achieved a high F-value
(i.e. 148.4) and a low P-value (< 0.0001), indicating its adequacy. It
can be used for the optimization of the culture medium’s EC and
growth rate parameters.
The results showed that the coefficients of the terms in the
model are statistically significant (P < 0.05). This confirms the supe-
riority of the model for the prediction of the EC reduction within
the variable ranges. A regression coefficient R2 of 0.98 was obtained
from the experimental data. The values of R2 Adjusted and R2 Predicted
were 0.98 and 0.96, respectively. These values were close to the R2
value, indicating a satisfactory model (Table 5).
According to Fig. 4 (A), the Lambda value of 0.66 would provide
the best power fit model. Fig. 4 (B) shows the excellent agreement
between the model predicted values and actual ones for culture
medium EC reduction at different salt concentrations.
Table 4 and Fig. 4 (C and D) show that the order of effective
parameters on salinity reduction by Chlorella vulgaris is salinity
followed by time and initial inoculum concentration. Particularly
according to Fig. 4 (C and D) by changing culture medium, the
salinity remained the most effective parameter. However, the sig-
nificance of this parameter was slightly reduced for the modified
medium (comparing Fig. 4 C with D). Based on the P-values and
F-values, it was found that the initial inoculum was the least impor-
tant parameter affecting the Chlorella vulgaris salinity reduction,
whereas the most important parameter was the salinity whose
removal enhanced the freshwater production.
3D surfaces and the contour plots of EC versus time and salinity
as well as EC versus time and initial inoculum are shown in Fig. 5.
The EC of BG11 culture medium was decreased by the elapse of
time at a specific salinity, as shown in Fig. 5 (A and B). After about
13 days, no significant reduction in EC was observed. In fact, at
the initial days of culturing, a significant decrease in EC was not
observed. The salt uptake occurred in two parts: first part due to
the increasing algal growth phase and the second part pertaining
to the stationary phase of algal growth. As their names imply, the
salt uptake rate is high during the growth phase, while very slight
removal occurs during the stationary phase.
However, EC reduction at different salinities was not signifi-
cantly impacted. At salinity of 0 up to 1000 ppm, the EC reduction
was concluded within 14 days. This reduction was only due to the
consumption of nutrient salts of the BG11 culture medium. It was
concluded that the desalination is not practical at salinities above
1000 ppm with microalgae in BG11 culture medium.
According to Fig. 5 (C and D) the variation in the initial inocu-
lum did not significantly affect the desiccation and EC reduction
process. It can be due to the culture medium type. As the Na+ and
Cl− ions were present among the salt recipe of BG11, in fact, the
amount of these ions was excessive to the added NaCl for the prepa-
ration of synthesized salt solutions with different concentrations.
This caused an increasing salinity effect on the microalgae cells to
be degraded sooner. Therefore, EC reduction did not occur while
decreasing the growth rate. As stated earlier, the Cl− and Na+ ions
present in the BG11 culture medium had two sources: (1) those in
the culture medium minerals and (2) the NaCl due to the salinity.
Therefore, the BG11 culture medium had a higher concentration
of Cl− and Na+ ions in comparison with MBG11. When the initial
inoculum came into contact with the BG11 culture medium, the
cells were exposed to a higher concentration of Na+ and Cl− ions,
causing their degradation. By degradation of these cells, no virtual
changes occurred in response to the increased microalgae growth
rate and EC reduction.
3D surfaces and the contour plots of EC versus time and salinity
as well as EC versus time and initial inoculum in the MBG11 culture
medium are illustrated in Fig. 6. This Figure confirms the superiority
of the MBG11culture medium due to its higher salinity removal
potential compared to the standard BG11 culture medium.
According to Fig. 6, by modifying the BG11 culture medium, the
EC reduction and consequently the desalination process enhance-
ment continued beyond 1000 ppm reaching about 5000 ppm
salinity. The excellent EC reduction by increasing the initial inocu-
lum is shown in Fig. 6 (C and D). According to Fig. 6, the highest
values of salinity reduction (SR) were predicted with the experi-
mental design when Chlorella vulgaris was cultivated at a salinity
of 3000 ppm during 12 days at 25 mL of initial inoculum per 100
mL of the culture medium.
After the identification of the terms affecting the salinity reduc-
tion, the experimental values were fitted to a power equation
obtained from multiple regression analysis, Eq. (3). The statisti-
cally insignificant terms at p > 0.05 were eliminated from the
model.
EC0.66 = 2.27 + 0.7(salinity)-0.24(time)-.08(initial inoculum)-
0.56(culture media)+0.04 (salinity × time)-0.16(salinity × culture
media)+0.13 (time × culture media)+0.05(initial inoculum × cul-
ture media)+0.04(salinity)2 +0.06(Time)2 +0.07(salinity)2 culture
media+0.03 (time)2 (3)

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Fig. 4. (A) The Box-Cox plot for power transforms, (B) Actual values versus the predicted values for the Chlorella vulgaris dry weight, (C) Order of effective parameters on
the growth of Chlorella vulgaris in BG11 culture media, (D) Order of effective parameters on the growth of Chlorella vulgaris in MBG11 culture media.
Eq. (3) was obtained for the light-dark cycle of 16−8 hours. After
obtaining the best conditions for the microalgae growth rate, a test
was conducted at the optimum conditions predicted by the RSM, to
verify the predicted results by comparing with compare the exper-
imental data. The final EC values obtained from the experimental
test and that predicted from RSM were 3.062 and 3.223 mS, respec-
tively. Therefore, the predicted results obtained from the proposed
model are in excellent agreement with the experimental ones, by
an acceptable error of 5%.
3.2.2. Effect of medium salinity on salinity reduction and growth
rate
To study the effect of salinity on EC reduction, the medium
salinity was increased from 1000–10000 ppm. Fig. 7 shows the
microalgae dry weight (DW) during 14 days at different salinities
in the range of 1000–10000 ppm for both culture media.
DW is a measure of the growth rate of microalgae. A high DW
means more growth of microalgae. So, a decrease in DW with
increasing salinity means a decrease in growth rate at high salin-
ity. According to Fig. 7, when the salinity increased, the microalgae
DW, which is a measure of growth rate, decreased. The growth of
microalgae and the rate of TDS removal were reduced by increas-
ing the medium salinity. The decline in TDS removal was due to a
decrease in algal growth. The increase in salt concentration marked
the end of the growth phase probably due to a sudden increase
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Process Safety and Environmental Protection xxx (xxxx) xxx–xxx
in the number of dead cells. In other words, microalgae growth
and TDS consumption were directly interdependent. However, the
reason for decreasing the microalgae growth, which resulted in a
decrease in TDS removal at higher salinities, should be explored
in the microalgae cellular mechanism. Increasing in the concentra-
tions of Na+ and Cl− occurred due to the increase in the amount
of osmotic pressure. Furthermore, for salinity values beyond the
tolerance level of the cells, the osmotic pressure imposed a more
rigid tolerance threshold of Na+ and Cl− pumps, resulting in cell
destruction (Eyster, 1958; Barrero-Gil et al., 2005; Arora et al.,
2019). Following cell destruction, cellular degradation occurred in
microalgae, leading to disruption of the cell division process and
reduction in growth rate.
3.3. Comparison of BG11 and MBG11 culture media
As stated earlier, the BG11 culture medium did not have accept-
able salinity reduction in comparison with the MBG11 medium.
To assure the better growth of Chlorella vulgaris in MBG11, it was
cultured in BG11 and MBG11 with no salinity. The Chlorella vulgaris
DW in BG11 and MBG11 non-saline culture media is shown in Fig. 8.
Generally, growth most microorganisms and microalgae such as
Chlorella vulgaris can be segregated into four phases which are lag
phase, log phase or exponential, stationary phase and finally death
phase (Guillard and Ryther, 1962; Das et al., 2011). Lag phase is the
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Fig. 5. The BG11 culture media: (A) 3D plot surface of EC versus time and salinity (B) the contour of EC versus time and salinity, (C) the 3D plot surface of EC versus time and
initial inoculum, (D) the contour of EC versus time and initial inoculum.
initial phase of cultivation in which the microalgae adapts to the
surrounding such as medium, pH, temperature and lighting. The
growth curve of Chlorella vulgaris used in this study also includes
these 4 steps. The first three days of their cultivation include the lag
phase. In fact, in the first three days, because the Chlorella vulgaris
enter the new culture medium, they first need time to get cus-
tomized to the fresh culture medium. On the other hand, because
in the early days, the culture medium contains a small number of
microalgae cells (it includes only inoculated cells), they grow slowly
and enter the second stage (exponential phase) at the end of the
third day. This is the reason of why DW of Chlorella vulgaris in this
study is almost hardly changed in the first three days. Because they
have been in their lag phase like other microorganisms that have
little growth.
But in generally, when the BG11 culture medium was modified,
the DW of Chlorella vulgaris, indicating the growth, was higher than
that of the BG11 culture medium. It can be due to the added K+ and
Ca2+ salts instead of Na+ salts in the MBG11 medium. The alter-
native salts were more effective on microalgae growth. Therefore,
the MBG11 culture medium was selected as the proper medium for
salinity reduction.
The EC versus the total DW at different salinities for both culture
media (BG11 and MBG11) is shown in Fig. 9. By the increased total
DW after 15 days, at the end of the desalination process, the EC
decreased significantly. The increase in total DW indicated more
microalgae growth rate. This growing rate resulted in the lower
EC, showing more salinity reduction. These results suggested that
the salt used in cellular uptake was mainly due to bioaccumu-
lation and the salt uptake was merely a function of the number
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Process Safety and Environmental Protection xxx (xxxx) xxx–xxx
of cells. Salinity reduction increased when the generation of new
cells was much higher than cells lost. Therefore, the MBG11 cul-
ture medium resulted in a significant decrease in EC as well as salt
removal.
The percentage of salinity reduction and Na removal at differ-
ent salinities is shown in Fig. 10. At 3000 ppm salinity, the salinity
reduction was 60 % with MBG11 culture medium, and the salinity
reduction was stopped for salinities above 5000 ppm. This shows
the potential of Chlorella vulgaris for desalination of brackish water.
The salinity reduction as a function of salinity for both culture
media is shown in Fig. 10 (A). By an increase in salinity up to
1000 and 2000 ppm, the salinity reduction efficiency in the BG11
culture medium was 45 % and 15 %, respectively. However, increas-
ing the salinity from 2000 to 10,000 ppm, the salinity reduction
was not significant for the BG11 culture medium. On the other
hand, in the MBG11 culture medium, salinity reductions of 85 %
and 65 % were obtained at salinities of 1000 ppm and 3000 ppm,
respectively. Increasing the salinity from 5000 to 10,000 ppm, no
appreciable salinity reduction was observed. This can be justified
by the fact that the cellular structure could result in bioaccumula-
tion by absorption of ionic contaminants on their structure. This is
an aerobic metabolic process which is done by giving energy from
a living organism (Velásquez and Dussan, 2009; Vijayaraghavan
and Yun, 2008). Metallic ions have been removed by bioaccumu-
lation of microalgae from water (Rangsayatorn et al., 2002; Gupta
and Rastogi, 2009; Vinod et al., 2010). The wall of microalgae cell
is mainly comprised of polysaccharides, lipids and proteins. The
wall of cell is also sticky and negatively charged, which permits
adsorption of cations (Rangsayatorn et al., 2002). The cation absorp-
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Fig. 6. The MBG11 culture media: (A) 3D plot surface of EC versus time and salinity, (B) the contour of EC versus time and salinity, (C) the 3D plot surface of EC versus time
and initial inoculum, (D) the contour of EC versus time and initial inoculum.

Fig. 8. The Chlorella vulgaris DW in BG11 and MBG11 culture media (non-saline).

Fig. 7. The effect of salinity on microalgae growth versus time (A) BG11 culture
media, (B) MBG11 culture media.

Fig. 9. The EC versus total DW at different salinities.

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Fig. 10. (A). The salinity reduction percentage versus salinity by Electrical con-
ductivity meter, (B) the Na removal percentage versus Na concentration by flame
photometry method.
Table 6
The Na content of chlorella vulgaris cell at 2000 ppm salinity.
Samples of 2000 ppm salinity Initial Na Content Final Na Content
base microalgae 0 0.1
treated microalgae after 16 days 0.1 23.3
deionized water for algae washing 0 5.4
tion mechanisms by Chlorella vulgaris included surface deposition,
physical and biosorption, active transport and passive diffusion
(Vinod and Rastogi, 2008). However, the primary channel through
which about 80 % of cations were absorbed was biosorption.
According to the results of the flame photometry (Fig. 10 (b)), by
measuring the amount of Na present in the culture media at initial
and final days of culturing process, the percentage of Na removal
was obtained at different salinities. These results confirmed the
trend of salinity removal by measuring the TDS. The removal effi-
ciencies were also confirmed by the atomic absorption test.
One possibility for the mechanism of salinity elimination by
microalgae as mentioned above in addition to its accumulation in
their cellular structure is the adsorption of Na+ on the microalgae
cell wall, having a negative charge. To investigate the validity of
this hypothesis, the salinity removal in the culture medium was
compared with the results obtained from algal sample flame pho-
tometry. It was prepared by separating the cells with centrifuge,
dissolving in deionized water and sonication prior to flame pho-
tometry measurement. An exemplary result for the concentration
of 2000 ppm in the modified BG11 medium is presented in Table 6.
According to Table 6, it can be seen that the amount of Na+
removed from the saline water was nearly the same as the amount
of Na+ adsorbed on the cell wall of Chlorella vulgaris. The slight dif-
ference seen in this comparison was due to washing the microalgae
cells with deionized water after centrifugation. During the washing
stage, some of the Na+ adsorbed on the surface of the microglial cell
were desorbed. To ensure the validity of this hypothesis, the Na+
content of the effluent water obtained from microalgae washing
was also analyzed by flame photometry tests. The results indicated
the presence of Na+ in deionized water obtained after microalgae
washing, validating the hypothesis.
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Fig. 11. The EC versus salinity in BG11 and MBG11 culture media in the initial and
final desalination process.
The remarkable point is that most of the removed Na+ was
maintained in the separated microalgae after washing, which may
indicate that the Na+ was fixed on the surface of the microalgae.
By stabilizing Na+ on microalgae, its biomass can be supplied for
applications requiring Na-rich biomass.
The EC versus salinity for the BG11 and MBG11 culture media is
shown in Fig. 11. According to Figs. 10 and 11, different behavior of
microalgae desalination in the MBG11 medium at the salinity range
of 0–5000 ppm can be observed. It means that the MBG11 culture
medium could shift the salinity reduction up to 5 times more than
that of the BG11. In other words, when the BG11 was modified,
the behavior of Chlorella vulgaris was different up to 5000 ppm of
salinity. It can be due to the MBG11 composition. The dissolved Na+
and Cl− ions were decreased in the MBG11 by removing Cl and Na
salts from the BG11 recipe. This resulted in a decrease of these ions
in the special salt concentration in the MBG11 compared with the
BG11. Beyond 5000 ppm salinity, the Chlorella vulgaris desalination
behavior was the same in both BG11 and MBG11 culture media.
As a result, the Chlorella vulgaris had a specific potential to
adsorb the Na+ and Cl− ions from the culture medium. Modifying
the culture medium, Chlorella vulgaris was forced to adsorb more
Na+ and Cl− from the salty water source. It was due to the elimina-
tion of other available salts containing Na+ or Cl− in their structure
by modification of BG11 culture medium, resulting in removing
more salt in comparison with the BG11 culture medium. Indeed,
by modification of BG11 culture medium, the desalination stopping
zone shifted from 1000–5000 ppm of salinity.
4. Conclusion
To study the suitability of using microalgae for brine desalina-
tion, at first, the adaptation of microalgae Chlorella vulgaris in saline
water was performed, and then the salt removal tests were con-
ducted. The live microalgae cells form the first step were separated
for use in the desalination process at the second step. The optimum
condition was determined through a parametric study using RSM
based on CCD. Culture medium, time, salinity and initial inoculum
were studied as the main parameters.
About 45 % salinity reduction in the BG11 culture medium
was achieved for salinities up to 1000 ppm. The BG11 culture
medium was modified to improve the desalination process. The
results showed that the algal growth and salt removal efficiency
enhanced in the MBG11 culture medium. The optimum values were
growth time of 12 days, 25 mL of initial inoculum per 100 mL of
culture media and 3000 ppm salinity. Moreover, the results indi-
cated that the removal efficiency for EC varied between 80−30%
for the MBG11 culture medium for a salinity range of 1000–5000
ppm. These results were obtained from the TDS measuring tests.
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Therefore, to assure the salinity removal, the Na content of culture
media and microalgae cells was also measured by flame photom-
etry which confirmed the TDS measuring finding. The Na+ content
measuring tests also indicated the intake salinity by Chlorella vul-
garis and fixation of the Na+ ions on cell wall of the microalgae. The
atomic absorption tests confirmed the obtained results, too.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared to
influence the work reported in this paper.
Acknowledgement
This work was supported by the vice chancellor of research and
technology of University of Isfahan under Grant No.942030237001.
References
Abdelmoez, W., Mahmoud, M.S., Farrag, T.E., 2014. Water desalination using humid-
ification/dehumidification (HDH) technique powered by solar energy: a detailed
review. Desalin. Water Treat. 52 (25–27), 4622–4640, http://dx.doi.org/10.1080/
19443994.2013.804457.
Ahmad, I., Hellebust, J.A., 1984. Osmoregulation in the Extremely Euryhaline Marine
Micro-Alga Chlorella autotrophica. Plant Physiol. 74 (4), 1010–1015, http://dx.
doi.org/10.1104/pp.74.4.1010.
Amezaga, J.M., Amtmann, A., Biggs, C.A., Bond, T., Gandy, C.J., Honsbein, A., et al.,
2014. Biodesalination: A case study for applications of photosynthetic bacteria in
water treatment. Plant Physiol. 164 (4), 1661–1676, http://dx.doi.org/10.1104/
pp.113.233973.
Arora, N., Kumari, P., Kumar, A., Gangwar, R., 2019. Delineating the molecular
responses of a halotolerant microalga using integrated omics approach to
identify genetic engineering targets for enhanced TAG production. Biotechnol.
Biofuels 12 (1), 2, http://dx.doi.org/10.1186/s13068-018-1343-1.
Barahoei, M., Hatamipour, M.S., Afsharzadeh, S., 2020a. CO2 capturing by chlorella
vulgaris in a bubble column photo-bioreactor; Effect of bubble size on CO2
removal and growth rate. J. Co2 Util. 37, 9–19, http://dx.doi.org/10.1016/j.jcou.
2019.11.023.
Barrero-Gil, J., Garciadeblas, B., Benito, B., 2005. Sodium, potassium-ATPases in algae
and oomycetes. J. Bioenerg. Biomembr 37, 269–278.
Bhargava, S., Saxena, R.K., Pandey, P.K., Bisen, P.S., 2003. Mutational engineering
of the cyanobacterium Nostoc muscorum for resistance to growth-inhibitory
action of LiCl and NaCl. Curr. Microbiol. 47 (1), 5–11, http://dx.doi.org/10.1007/
s00284-002-3921-4.
Blumwald, E., Mehlhorn, R.J., Packer, L., 1983. Ionic Osmoregulation during Salt
Adaptation of the Cyanobacterium Synechococcus 6311. Plant Physiol. 73 (2),
377–380, http://dx.doi.org/10.1104/pp.73.2.377.
Bohn, P.W., Elimelech, M., Georgiadis, J.G., Mariñas, B.J., Mayes, A.M., Mayes,
A.M., 2009. Science and technology for water purification in the coming
decades. Nat. Nanotechnol. 452 (March), 337–346, http://dx.doi.org/10.1142/
9789814287005 0035.
Das, P., Aziz, S.S., Obbard, J.P., 2011. Two phase microalgae growth in the open system
for enhanced lipid productivity. Renew. Energy 36 (9), 2524–2528, http://dx.doi.
org/10.1016/j.renene.2011.02.002.
de Lacerda, L.P., Lange, L.C., Costa França, M.G., Zonta, E., 2015. Salinity Reduction and
Biomass Accumulation in Hydroponic Growth of Purslane (Portulaca oleracea).
Int. J. Phytoremediation 17 (3), 235–241, http://dx.doi.org/10.1080/15226514.
2014.883494.
Ditta, A., 2016. Salt Tolerance in Cereals: Molecular Mechanisms and Applications.
In: Molecular Stress Physiology of Plants. Springer, India, pp. 133–154 https://
link.springer.com/book/10.1007/978-81-322-0807-5.
El Sergany, F.A.R., El Fadly, M., El Nadi, M.H.A., 2014. Brine desalination by using
algae ponds under nature conditions. Am. J. Environ. Eng. 4 (4), 75–79, http://
dx.doi.org/10.11648/j.wros.20140306.11.
El Nadi, M.H., El Sergany, F.A.G.H., 2010. Water desalination by algea. ASU J. Civil
Eng. 2, 105–114.
El Nadi, M.H.A., El Sergany, F.A.G.H., El Hosseiny, E.H., 2014. Desalination using algae
ponds under nature egyptian conditions. J. Water Resour. Ocean. Sci. 3 (6), 69–73,
http://dx.doi.org/10.11648/j.wros.20140306.11.
Espie, G.S., Miller, A.G., Canvin, D.T., 1988. Characterization of the Na + -Requirement
in Cyanobacterial Photosynthesis. Plant Physiol. 88 (3), 757–763, http://dx.doi.
org/10.1104/pp.88.3.757.
Eyster, C., 1958. Chloride effect on the growth of Chlorella pyrenoidosa. Nature 181
(4616), 1141–1142, http://dx.doi.org/10.1038/1811141a0.
Falhof, J., Pedersen, J.T., Fuglsang, A.T., Palmgren, M., 2016. Plasma membrane H+-
ATPase regulation in the center of plant physiology. Mol. Plant 9 (3), 322–337,
http://dx.doi.org/10.1016/j.molp.2015.11.002.
247
Process Safety and Environmental Protection xxx (xxxx) xxx–xxx
Fisher, M., Gokhman, I., Pick, U., Zamir, A., 1997. A structurally novel transferrin-
like protein accumulates in the plasma membrane of the unicellular green alga
Dunaliella salina grown in high salinities. J. Biol. Chem. 272 (3), 1565–1570,
http://dx.doi.org/10.1074/jbc.272.3.1565.
Ghalavand, Y., Hatamipour, M.S., Rahimi, A., 2015. A review on energy consumption
of desalination processes. Desalin. Water Treat. 54 (6), 1526–1541, http://dx.
doi.org/10.1080/19443994.2014.892837.
Gimmler, H., Wiedemann, C., Moller, E.M., 1981. The metabolie response ofthe
halotolerant green alga Dllllaliella parva to hypertonie shocks. Berichte der
Deutschen Botanischen Gesellschaft 94, 613–634.
Guillard, R.R.L., Ryther, J.H., 1962. Studies of marine planktonic diatoms. I. Cyclotella
nana hustedt, and Detonula confervacea (CLEVE). Can. J. Microbiol. 8, 229–239,
Can. J. Microbiol., 8(1140), 229–239.
Gupta, V.K., Rastogi, A., 2009. Biosorption of hexavalent chromium by raw and acid-
treated green alga Oedogonium hatei from aqueous solutions. J. Hazard. Mater.
163 (1), 396–402, http://dx.doi.org/10.1016/j.jhazmat.2008.06.104.
Hare, P.D., Cress, W.A., Van Staden, J., 1998. Dissecting the roles of osmolyte accu-
mulation during stress. Plant Cell Environ. 21 (6), 535–553.
Hasegawa, P.M., Bressan, R.A., Zhu, J.K., Bohnert, H.J., 2000. Plant cellular and molec-
ular responses to high salinity. Annu. Rev. Plant Biol. 51 (1), 463–499.
Hiremath, S., Mathad, P., 2010. Impact of Salinity on the Physiological and Biochem-
ical Traits of Chlorella vulgaris Beijerinck. J. Algal Biomass Util. 1 (2), 51–59.
Ho, S.H., Chen, C.Y., Lee, D.J., Chang, J.S., 2011. Perspectives on microalgal CO2-
emission mitigation systems - A review. Biotechnol. Adv. 29 (2), 189–198, http://
dx.doi.org/10.1016/j.biotechadv.2010.11.001.
Hong, B., Barg, R., Ho, T.H.D., 1992. Developmental and organ-specific expression of
an ABA-and stress-induced protein in barley. Plant Mol. Biol. 18 (4), 663–674.
Hoque, M.A., Okuma, E., Banu, M.N.A., Nakamura, Y., Shimoishi, Y., Murata, Y., 2007.
Exogenous proline mitigates the detrimental effects of salt stress more than
exogenous betaine by increasing antioxidant enzyme activities. J. Plant Physiol.
164 (5), 553–561.
Iaccarino, M., 2019. Water, population growth and contagious diseases. Water
(Switzerland) 11 (2), http://dx.doi.org/10.3390/w11020386.
Jlassi, A., Zorrig, W., Khouni, A.El., Lakhdar, A., Smaoui, A., Abdelly, C., Rabhi, M.,
2013. Phytodesalination of a Moderately-Salt-Affected Soil By Sulla Carnosa.
Int. J. Phytoremediation 15 (4), 398–404, http://dx.doi.org/10.1080/15226514.
2012.716104.
Kalleary, S., Mohammed Abbas, F., Ganesan, A., Meenatchisundaram, S., Srinivasan,
B., Packirisamy, A.S.B., et al., 2014. Biodegradation and bioelectricity generation
by microbial desalination cell. Int. Biodeterior. Biodegradation 92, 20–25, http://
dx.doi.org/10.1016/j.ibiod.2014.04.002.
Morillo, J., Usero, J., Rosado, D., El Bakouri, H., Riaza, A., Bernaola, F.J., 2014.
Comparative study of brine management technologies for desalination plants.
Desalination 336 (1), 32–49, http://dx.doi.org/10.1016/j.desal.2013.12.038.
Narayan, G.P., McGovern, R.K., Zubair, S.M., Lienhard, J.H., 2012. High-temperature-
steam-driven, varied-pressure, humidification-dehumidification system cou-
pled with reverse osmosis for energy-efficient seawater desalination. Energy
37 (1), 482–493, http://dx.doi.org/10.1016/j.energy.2011.11.007.
Nie, J., Sun, Y., Zhou, Y., Kumar, M., Usman, M., Li, J., et al., 2020. Bioremedia-
tion of water containing pesticides by microalgae: mechanisms, methods, and
prospects for future research. Sci. Total Environ. 707, 136080, http://dx.doi.org/
10.1016/j.scitotenv.2019.136080.
Pires, J.C.M., Alvim-Ferraz, M.C.M., Martins, F.G., Simões, M., 2012. Carbon dioxide
capture from flue gases using microalgae: Engineering aspects and biorefinery
concept. Renewable Sustainable Energy Rev., http://dx.doi.org/10.1016/j.rser.
2012.02.055.
Priyadarshani, I., Rath, B., 2012. Commercial and industrial applications of micro
algae – a review. J. Algal Biomass Utln. 3 (4), 89–100.
Rabhi, M., Ferchichi, S., Jouini, J., Hamrouni, M.H., Koyro, H.W., Ranieri, A., et al., 2010.
Phytodesalination of a salt-affected soil with the halophyte Sesuvium portula-
castrum L. to arrange in advance the requirements for the successful growth of
a glycophytic crop. Bioresour. Technol. 101 (17), 6822–6828, http://dx.doi.org/
10.1016/j.biortech.2010.03.097.
Rangsayatorn, N., Upatham, E.S., Kruatrachue, M., Pokethitiyook, P., Lanza, G.R., 2002.
Phytoremediation potential of Spirulina (Arthrospira) platensis: Biosorption and
toxicity studies of cadmium. Environ. Pollut. 119 (1), 45–53, http://dx.doi.org/
10.1016/S0269-7491(01)00324-4.
Sahle-Demessie, E., Aly Hassan, A., El Badawy, A., 2019. Bio-desalination of brack-
ish and seawater using halophytic algae. Desalination 465 (January), 104–113,
http://dx.doi.org/10.1016/j.desal.2019.05.002.
Sharon, H., Reddy, K.S., 2015. A review of solar energy driven desalination technolo-
gies. Renewable Sustainable Energy Rev., http://dx.doi.org/10.1016/j.rser.2014.
09.002.
Spiegler, K.S., El-Sayed, Y.M., 2001. The energetics of desalination processes. Desali-
nation 134 (1–3), 109–128, http://dx.doi.org/10.1016/S0011-9164(01)00121-
7.
Taheri, R., Razmjou, A., Szekely, G., Hou, J., Ghezelbash, G.R., 2016. Biodesalina-
tion - On harnessing the potential of nature’s desalination processes. Bioinspir.
Biomim., http://dx.doi.org/10.1088/1748-3190/11/4/041001.
Thomas, J., Apte, S.K., 1984. Sodium requirement and metabolism in nitrogen-fixing
cyanobacteria. J. Biosci. 6 (5), 771–794, http://dx.doi.org/10.1007/BF02702719.
Velásquez, L., Dussan, J., 2009. Biosorption and bioaccumulation of heavy metals
on dead and living biomass of Bacillus sphaericus. J. Hazard. Mater. 167 (1–3),
713–716, http://dx.doi.org/10.1016/j.jhazmat.2009.01.044.
Vijayaraghavan, K., Yun, Y.S., 2008. Bacterial biosorbents and biosorption. Biotech-
nol. Adv. 26 (3), 266–291, http://dx.doi.org/10.1016/j.biotechadv.2008.02.002.
G Model
PSEP-2512; No. of Pages 248 ARTICLE IN PRESS
M. Barahoei et al. Process Safety and Environmental Protection xxx (xxxx) xxx–xxx

Vinod, K., Rastogi, A., 2008. Biosorption of lead(II) from aqueous solutions by non- Zuppini, A., Gerotto, C., Baldan, B., 2010. Programmed cell death and adaptation: two
living algal biomass Oedogonium sp. and Nostoc sp.-A comparative study. different typesof abiotic stress response in a unicellular chlorophyte. Plant Cell
Colloids Surfaces B: Biointerfaces 64 (2), 170–178, http://dx.doi.org/10.1016/ Physiol. 51 (6), 884–895.
j.colsurfb.2008.01.019.
Vinod, K., Rastogi, A., Nayak, A., 2010. Biosorption of nickel onto treated alga (Oedo-
gonium hatei): Application of isotherm and kinetic models. J. Colloid Interface
Sci. 342 (2), 533–539, http://dx.doi.org/10.1016/j.jcis.2009.10.074.

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