membranes

Article
Nutrient Removal and Membrane Performance of an Algae
Membrane Photobioreactor in Urban Wastewater Regeneration
Verónica Díaz 1 , Laura Antiñolo 2 , José Manuel Poyatos Capilla 2 , Mari Carmen Almécija 1 ,
María del Mar Muñío 1 and Jaime Martín-Pascual 2, *

1 Department of Chemical Engineering, University of Granada, 18071 Granada, Spain

2 Department of Civil Engineering, University of Granada, 18071 Granada, Spain
* Correspondence: [email protected]; Tel.: +34-958-24-61-55

Abstract: The increase in industry and population, together with the need for wastewater reuse,
makes it necessary to implement new technologies in the circular economy framework. The aim of
this research was to evaluate the quality of the effluent of an algae membrane photobioreactor for the
treatment of the effluent of an urban wastewater treatment plant, to characterise the ultrafiltration
membranes, to study the effectiveness of a proposed cleaning protocol, and to analyse the perfor-
mance of the photobioreactor. The photobioreactor operated under two days of hydraulic retention
times feed with the effluent from the Los Vados wastewater treatment plant (WWTP) (Granada, Spain).
The microalgae community in the photobioreactor grew according to the pseudo-second-order model.
The effluent obtained could be reused for different uses of diverse quality with the removal of total
nitrogen and phosphorus of 56.3% and 64.27%, respectively. The fouling of the polyvinylidene difluo-
ride ultrafiltration membrane after 80 days of operation was slight, increasing the total membrane
resistance by approximately 22%. Moreover, the higher temperature of the medium was, the lower
Citation: Díaz, V.; Antiñolo, L.;
intrinsic resistance of the membrane. A total of 100% recovery of the membrane was obtained in the
Poyatos Capilla, J.M.; Almécija, M.C.;
two-phase cleaning protocol, with 42% and 58%, respectively.
Muñío, M.d.M.; Martín-Pascual, J.
Nutrient Removal and Membrane
Performance of an Algae Membrane
Keywords: fouling; ultrafiltration membrane; microalgae; photobioreactor; wastewater reuse
Photobioreactor in Urban Wastewater
Regeneration. Membranes 2022, 12,
982. https://doi.org/10.3390/
membranes12100982 1. Introduction
Academic Editors: Alfredo Cassano
Microalgae are currently some of the most promising renewable raw materials to
and Dimitra Banti
produce different subproducts [1–3]. From both an economic and environmental point
of view, microalgae are attractive because their cultivation and processing could involve
Received: 2 September 2022 wastewater treatment and carbon dioxide capture from combustion gases [4–6]. Currently,
Accepted: 6 October 2022 microalgae-based systems are some of the most promising technologies for tertiary treat-
Published: 10 October 2022
ment in wastewater treatment plants (WWTPs). This is due to their associated low costs
Publisher’s Note: MDPI stays neutral and environmental advantages [7,8].
with regard to jurisdictional claims in Nowadays, microalgae cultivation has a high cost in terms of the nutrients needed.
published maps and institutional affil- Therefore, the use of wastewater may be a viable solution because of its nutrient content,
iations. coupled with the ability of these micro-organisms to tolerate very wide conditions of tem-
perature, pH, or salinity [9,10]. Wastewater provides the nutrients necessary for the growth
of the microalgae, in turn reducing the cost of production and greenhouse gas emissions
because the microalgae will absorb some of the carbon generated in the wastewater treat-
Copyright: © 2022 by the authors.
ment that would be released as carbon dioxide [6,9,11]. This achieves the requirements of
Licensee MDPI, Basel, Switzerland.
wastewater discharge standards, which specify very low nutrient concentrations to avoid
This article is an open access article
eutrophication [12].
distributed under the terms and
Wastewater treatment with microalgae reduces the costs of microalgae biomass produc-
conditions of the Creative Commons
tion by 40% of the total cost, generating a lower energy demand for nutrient removal and,
Attribution (CC BY) license (https://
therefore, presenting lower costs compared to conventional wastewater treatment [13–15].
creativecommons.org/licenses/by/
4.0/).
On one hand, microalgae can use inorganic phosphorus and nitrogen for their growth,

Membranes 2022, 12, 982. https://doi.org/10.3390/membranes12100982 https://www.mdpi.com/journal/membranes

Membranes 2022, 12, 982 2 of 16

as well as produce oxygen during photosynthesis. On the other hand, microalgae are
also used for the removal of coliforms because the environmental growth conditions for
microalgae are unfavourable for these microorganisms [16]. Both phosphorus and nitrogen
can be considered limiting factors in the growth of microalgae [17].
Much of the literature on the use of microalgae for wastewater treatment focuses on
the treatment of synthetic wastewater using pure microalgae strains, ignoring bacteria
and other microorganisms present in real wastewater. For this reason, reproducibility
from laboratory to real WWTP conditions is complicated. In WWTPs, a heterogeneous
and complex consortia of microalgae, cyanobacteria, and other microorganisms would be
used [18]. Therefore, we decided to work with real treated urban wastewater in this study.
It is known that biological wastewater treatment with activated sludge is energy-
intensive. Therefore, the use of bacterial and microalgae consortia could be an alternative
treatment method [19]. In this way, aeration costs could be saved due to the algae’s ability
to produce sufficient oxygen for bacterial growth and to adsorb ammonium and phos-
phate [20], just as the bacteria provide carbon dioxide for the growth of the microalgae [21].
Some of the microalgae used for wastewater treatment belong to the genera Scenedesmus sp.,
Chlorella sp., Desmodesmus sp., Neochloris sp., Chlamydomonas spp., and Nitzschia spp. [7,22].
One of the most promising technologies widely used for municipal and industrial
wastewater treatment is the membrane photobioreactor (MPBR), due to its advantages in
terms of small footprint, high quality of treated water, and easy operation [23,24].
Membranes are a very promising technology for harvesting microalgae cultures [25].
This is due to the energy and cost savings compared to using centrifuges, as well as
the fact that they retain almost all of the biomass [12]. According to other studies, the
microalgae concentration in a photobioreactor without a membrane is significantly lower
than in an MPBR [12,26]. In addition, membrane filtration in the photobioreactor prevents
the microalgae culture from being washed out, as the solid retention time (SRT) in the
photobioreactor will be different from the hydraulic retention time (HRT). In this way,
higher yields and biomass concentrations are achieved [12,27,28]. This also affects the
concentration of nutrients, especially when working with domestic wastewater, in which
nitrogen and phosphorus concentrations are relatively low [29]. Another advantage of using
membranes in photobioreactors is the reduction in volume, as the removal of nutrients
present in urban wastewater would require less space [12]. However, a potential drawback
of MPBRs is that if the effluent is untreated wastewater, it can lead to the death of the
microalgae species being cultivated in the effluent, in which case it is necessary to design
an appropriate pre-treatment. In the same way, it is important to correctly select the species
of microalgae to be cultivated, as not all of them are capable of adapting to the conditions
of the wastewater [30]. Another disadvantage of these photobioreactors is the high risk of
contamination of the microalgae culture, as well as the tedious and costly work involved in
harvesting the microalgae [31].
Previous authors have claimed that the maintenance and operating costs of an MPBR
are almost eight times lower than those of conventional nutrient removal treatments [32].
Even the energy costs associated with an MPBR would be lower than those used in closed
photobioreactors or open ponds [30]. According to other authors, the potential of MPBRs
for domestic wastewater treatment using a consortium of bacteria and microalgae has not
yet been studied [20].
In the present investigation, the continuous performance of a photobioreactor equipped
with a polyvinylidene fluoride (PVDF) ultrafiltration membrane for the treatment of urban
wastewater already treated with microalgae with a two-day HRT was studied to achieve
the following objectives: (1) evaluation of the water quality of the effluent obtained in a
photobioreactor equipped with an ultrafiltration membrane for the treatment of already
treated urban wastewater from a WWTP using microalgae culture, (2) analysis of the per-
formance of the photobioreactor, and (3) evaluation of the operation of the ultrafiltration
membrane in a microalgae culture.
 2.1. Pilot plant
This study was carried out in a cylindrical methacrylate photobioreactor of 18 L (Fig-
ure 1). It was equipped with a hollow-fibre polymeric ultrafiltration membrane (PVDF)
configured according to the needs of the plant, and manufactured from a reconditioned
Membranes 2022, 12, 982 membrane with a larger unit surface area (0.92 m2), a nominal pore size of 0.04 μm, and 3 of a
16
membrane life of approximately 10 years.
The fibres of the ultrafiltration membrane were taken and divided into four smaller
filtration modules, adapted to the photobioreactor used in this research. To this end, the
2. Materials and Methods
fibres that make up the new module were sealed with silicone to the PVC parts that allow
2.1. Pilot Plant
the membrane to be connected to the filtration system. The length of each hollow fibre
was 500This
mm, studyandwas carried
the total out in asurface
membrane cylindrical
area ofmethacrylate photobioreactor
this new membrane of2. 18 L
was 0.2 m
(Figure
The permeate was extracted through the membrane by a suction-backwashing(PVDF)
1). It was equipped with a hollow-fibre polymeric ultrafiltration membrane peri-
staltic pumpaccording
configured to the needs ofPumps
(323 U, Watson-Marlow the plant, and Marlow,
Group, manufactured
UK). Thefrompermeate
a reconditioned
extrac-
membrane with a larger unit surface 2 ), a nominal pore size of 0.04 µm, and a
tion and backwashing periods were 9 area
and 1(0.92
min,mrespectively.
membrane life of approximately 10 years.

Figure
Figure1.1.Experimental
Experimentalpilot
pilotplant
plantused
usedininthe
thelaboratory.
laboratory.

The
To fibres ofand
automate the control
ultrafiltration membrane
the correct were
operation of taken and divided into
the photobioreactor, four smaller
a vacuum and
filtration modules, adapted to the photobioreactor used in this research.
pressure gauge (Potermic, Barcelona, Spain) were installed to control the pressure during To this end, the
fibres that make up the new module were sealed with silicone to the PVC
the filtering process and level probes to keep the photobioreactor at an approximately parts that allow
the membrane
constant volume. to In
be addition,
connectedthe to the filtration system.
photobioreactors Thea length
have of each
continuous hollowsystem
stirring fibre was
of
500 mm, and the total membrane surface area of this new membrane was 0.2 m 2.
approximately 8 Hz (MSL 63 A, Cime Motors, Girona, Spain) and continuous air injection
The permeate
by means wasdiffuser
of a circular extracted through
at the base ofthethe
membrane by a suction-backwashing
photobioreactor. In addition to naturalperi-
staltic pump (323 U, Watson-Marlow Pumps Group, Marlow, UK). The permeate extraction
lighting, the photobioreactor was illuminated by fluorescent lights (L36 W/868, OSRAM
and backwashing periods were 9 and 1 min, respectively.
DS, Munich, Germany) placed around the plant, providing an average irradiance in the
To automate and control the correct operation of the photobioreactor, a vacuum and
photobioreactor of 13.24 μmol/m2·s. A 12/12-h light/dark cycle was established by means
pressure gauge (Potermic, Barcelona, Spain) were installed to control the pressure during
of a timer.
the filtering process and level probes to keep the photobioreactor at an approximately
constant volume. In addition, the photobioreactors have a continuous stirring system of
approximately 8 Hz (MSL 63 A, Cime Motors, Girona, Spain) and continuous air injection
by means of a circular diffuser at the base of the photobioreactor. In addition to natural
lighting, the photobioreactor was illuminated by fluorescent lights (L36 W/868, OSRAM
DS, Munich, Germany) placed around the plant, providing an average irradiance in the
photobioreactor of 13.24 µmol/m2 ·s. A 12/12-h light/dark cycle was established by means
of a timer.
To determine the average irradiance (light intensity) in the photobioreactor, an HD
2303.2 Delta OHM light meter (Arava, Spain) was used with the DLP471 PAR probe
(Arava, Spain) for photon measurement. Measurements were taken on and around the
photobioreactor surface, both perpendicular and parallel to the photobioreactor.
The photobioreactor was continuously fed with treated urban wastewater from real
wastewater taken from WWTP of Los Vados (Granada, Spain), and naturally occurring
Membranes 2022, 12, 982 4 of 16

microalgae species were cultured. The HRT set in the photobioreactor was two days,
where the transmembrane operating pressure (TMP) was approximately 0.01 bar. The
experimental cycle lasted 80 days, coinciding with the SRT.

2.2. Experimental Procedure

Samples were taken on alternate days in the influent, photobioreactor, and efflu-
ent. Once the samples were collected, the pH was measured with a Crison pH 25 meter
(Hach, Barcelona, Spain), and the temperature and conductivity were measured using the
multifunctional meter PCE-PHD 1 (PCE Ibérica SL, Alicante, Spain). For the analysis of
Total Suspended Solids (TSS) present in the sample, 0.45 µm Millipore glass fibre filters
were used following standard methods [33]. The filtrate was used for the analysis and
quantification of nutrients in the samples, determined by ion chromatography using a
conductivity detector (Metrohm, Metrohm AG, Herisau, Switzerland). Finally, the Heλios
spectrophotometer (Thermo Spectronic, Madrid, Spain) was used to measure turbidity at a
wavelength of 650 nm and the optical density of the culture at 680 nm, following the same
procedure as other authors [8,34].

2.3. Kinetic Modelling

Table 1 shows the six kinetic models that were evaluated to adjust the kinetics of mi-
croalgae biomass growth in the photobioreactor, measured as TSS throughout the evolution
of the concentration over the time (Ct ):

Table 1. Kinetic models.

Kinetic Model Equation

Pseudo-first-order lnCt = lnC0 − kt
Pseudo-second-order Ct = C0 − 1 t t
( h + Ce )
Zero-order Ct = C0 + k0 t
First-order ln(Ct ) = lnC0 + k1 t
Second-order 1 1
Ct = C0 + k 2 t
1
Third-order
C2
= C12 + k3 t
t 0

C0 (g·L−1 ) represents the initial biomass concentration, Ct (g·L−1 ) is the biomass

concentration at any time, and Ce (g·L−1 ) is the biomass equilibrium concentration. The
experimental time is expressed as t (days) and the model constant as k (days−1 ). The
parameter h relates the biomass concentration at equilibrium and the kinetic model constant
as follows: h = k ·Ce2 (g2 ·L−2 ·days−1 ).
The data were fitted to the models described above, minimising the sum of squared er-
rors between the empirical and modelled data using the Microsoft Excel Solver application.

2.4. Membrane Filtration Tests

The membrane was characterised prior to use to determine the intrinsic resistance of
the clean membrane. For this purpose, a test was carried out with clean water. Subsequently,
the behaviour of the clean membrane was studied with the content of the photobioreactor
(microalgae culture in treated urban wastewater).
At the end of the study cycle, the membrane was again characterised with water and
the content of the photobioreactor. In this way, the total resistance of the membrane after
fouling was obtained.
Finally, the recovery of the membrane permeability was studied by means of a two-
stage cleaning protocol. In the first stage of the cleaning protocol, the membrane was
inorganically cleaned with citric acid (800 ppm) for 5 h. The second cleaning step consisted
of an organic cleaning with sodium hypochlorite (800 ppm) for 5 h. After each of the
cleaning stages, the membrane was characterised in water to evaluate the cleaning efficiency
of each stage.
Membranes 2022, 12, 982 5 of 16

Each of the tests was carried out at three different temperatures (20, 25, and 30 ◦ C). To
carry them out, the volumes obtained after 30 s of filtration at different values of TMP were
recorded. In this way, filtration fluxes were obtained for each of the TMP sets.

2.4.1. Membrane Characterisation

The membrane permeability (K) was calculated from the following equation as de-
scribed above [35], where J is the permeate flux (LHM, litres per square metre per hour
filtered) and TMP is the measured transmembrane pressure (bar):

TMP TMP
J = K · TMP = = 0 (1)
µ( R M + RCP + R F ) R M + R0F

RM represents the intrinsic resistance of the membrane, RCP is the resistance that occurs
in the first moments of filtration, which is reversible, and RF corresponds to the resistance
that occurs at longer times, which corresponds to fouling. RM 0 and RF 0 represent the total
value of the intrinsic membrane resistance and the total value of the fouling resistance,
respectively, by integrating the viscosity constant for each temperature.
This permeability constant encompasses the influence of temperature and the intrinsic
resistance of the membrane. On the other hand, the intrinsic membrane resistance (RM 0 ) was
calculated as the inverse of the permeability, which had been obtained with the membrane
cleaned in water:
1
RM0 = (2)
K
The fouling resistance (RF 0 ) of the membrane was calculated as described in Equation (3),
where Ri 0 represents the total membrane resistance in each of the cases studied.

R F 0 = Ri 0 − R M 0 (3)

2.4.2. Recovery of Membrane Characteristics

The efficiency of the membrane cleaning was calculated as shown in Equation (4),
where R0 represents the total membrane resistance with a soiled membrane, Ri 0 represents
the total membrane resistance in each of the cases studied, R0 i−1 represents the total
membrane resistance of the case before the one under study, and RM 0 is the intrinsic
membrane resistance:
R 0 − Ri 0
E (% ) = i −1 ·100 (4)
R0 − R M 0

2.4.3. Analysis of the Operational TMP

The TMP was determined by setting the flow rate necessary for the HRT in the
photobioreactor of the treated urban wastewater to be two days. It was verified that the
operating TMP of the membrane was in the pressure-controlled working region.
To evaluate the flux at different TMPs using the contents of the photobioreactor, tests
were carried out with both clean and dirty membranes after the end of the experimental
cycle in the photobioreactor. For this purpose, the equation of the series resistance model
described below was used:
TMP
J= 0 (5)
R M + b· TMP
The parameter b is a constant which relates the behaviour of the TMP to the matter
transfer in the filtration process.

3. Results and Discussion

3.1. Photobioreactor Operation
After analyses of the samples were taken, the average operating conditions of the
reactor during the experimental period were 7.40 ± 0.88 for pH, 15.54 ± 0.89 ◦ C for
temperature, and 954.30 ± 75.82 µS/cm for conductivity.
 transfer in the filtration process.

3. Results and Discussion

3.1. Photobioreactor Operation
Membranes 2022, 12, 982 6 of 16
After analyses of the samples were taken, the average operating conditions of the
reactor during the experimental period were 7.40 ± 0.88 for pH, 15.54 ± 0.89 °C for temper-
ature, and 954.30 ± 75.82 μS/cm for conductivity.
Figure
Figure2a 2ashows
showsthe theevolution
evolution of of
thethe
TSS in the
TSS photobioreactor
in the during
photobioreactor the experimen-
during the experi-
tal time.time.
mental The concentration
The concentrationof algal biomass
of algal increases
biomass as time
increases passes
as time until
passes thethe
until stationary
station-
phase is reached.
ary phase In this
is reached. case
In this thethe
case results obtained
results obtainedduring
duringthe
thegrowth
growthphase
phasehavehavebeen
been
represented up to their maximum point, where the stationary phase,
represented up to their maximum point, where the stationary phase, not represented,not represented,
would
wouldbegin.
begin.
0.20
800
0.18
700
0.16
0.14 600

TURBIDITY (NTU)
0.12 500
TSS (g/L)

0.10 400
0.08
300
0.06
200
0.04
0.02 100
0.00 0
0 20 40 60 80 0 20 40 60 80
Experimental time (days) Experimental time (days)
(a) (b)
Figure 2. Evolution of TSS (a) and turbidity (b) in the photobioreactor during the experimental period.
Figure 2. Evolution of TSS (a) and turbidity (b) in the photobioreactor during the experimental
period.
In the same way, the turbidity results obtained in the photobioreactor are shown in
Figure 2b and
In the same expressed
way, thein NTU (Nephelometric
turbidity results obtainedturbidity unit), which are are
in the photobioreactor related to the
shown in
TSS concentration in the photobioreactor, increasing as the concentration of
Figure 2b and expressed in NTU (Nephelometric turbidity unit), which are related to themicroalgae
biomass
TSS increases.in the photobioreactor, increasing as the concentration of microalgae
concentration
Figure
biomass 3 shows the evolution of the optical density of the microalgae culture. These
increases.
values were3 obtained
Figure shows thebyevolution
taking the difference
of the optical between
density ofthe
theabsorbance
microalgaemeasurement
culture. These at
680 nmwere
values
Membranes 2022, 12, x FOR PEER REVIEW of the microalgae
obtained photobioreactor
by taking culture
the difference and the
between themeasurement obtained for the
absorbance measurement at 7 of
microalgae
680 nm of the feed (treated urban
microalgae wastewater).
photobioreactor culture and the measurement obtained for the
microalgae feed (treated urban wastewater).

0.8

0.7

0.6
Optical density (OD)

0.5

0.4

0.3

0.2

0.1

0
0 20 40 60 80
Experimental time (days)

Figure 3. Evolution of optical density (OD) in the photobioreactor during the experimental period.
Figure 3. Evolution of optical density (OD) in the photobioreactor during the experimental period

The same growth trend was observed in the three parameters studied, with a diffe
ence—in the study of the optical density, the interferences that could be caused by the TS
contained in the treated urban wastewater, bacteria, and other microorganisms that influ
ence the correct evaluation of the growth of the algal biomass were eliminated. Figure
 0.1

0
0 20 40 60 80
Experimental time (days)
Membranes 2022, 12, 982 7 of 16

Figure 3. Evolution of optical density (OD) in the photobioreactor during the experimental period.

The same
samegrowth
growthtrend
trendwas
wasobserved
observedin in
thethe
three parameters
three studied,
parameters withwith
studied, a difference—
a differ-
in the study
ence—in of theofoptical
the study density,
the optical the the
density, interferences that
interferences could
that bebecaused
could causedbybythe
the TSS
urban wastewater,
contained in the treated urban wastewater, bacteria, and
and other
other microorganisms
microorganisms thatthat influ-
influ-
ence the correct evaluation of the growth of the algal biomass
algal biomass were eliminated.
eliminated. Figure 4
shows the microalgae present in the photobioreactor, which grew naturally in the
shows the microalgae present in the photobioreactor, which grew naturally in the treated treated
urban wastewater.
urban wastewater.

(a) (b)
Figure 4. (a) Microscope photograph of photobioreactor contents. (b) Enlarged microscope photograph of microalgae cul-
Figure 4. (a) Microscope photograph of photobioreactor contents. (b) Enlarged microscope photo-
ture.
graph of microalgae culture.
The kinetic constants
The kinetic constants and
and correlation
correlation rates
rates obtained
obtained for each of
for each of the
the kinetic
kinetic models
models
evaluated
evaluated for
for the
the concentration
concentration of
of TSS
TSS and
and for
for the
the optical
optical density
density obtained
obtained in
in the
the photo-
photo-
bioreactor,
bioreactor, which has been evaluated at a wavelength of 680 nm, are presented in Table
which has been evaluated at a wavelength of 680 nm, are presented in 2:
Table 2:

Table 2.
Table 2. Kinetic
Kinetic constants
constants and
and correlation
correlation rates
rates for
for each
each evaluated
evaluated kinetic
kineticmodel.
model.

Total Suspended Solids (TSS) Optical Density (OD)

Total Suspended Solids (TSS) Optical Density (OD)
Kinetic Model Kinetic
Model Constants Correlation Rate R2 Correlation
Model Constants CorrelationCorrela-
Rate R2
Model Model Constants Model Constants tion Rate
Pseudo-first order k = −3.677·10 days−1
−2 0.9444 Rate
k=− R1.243
2 ·10−1 days−1 0.7897 2 R
Ce = 1.075 g/L Ce = 2.316 g/L
Pseudo-second order Pseudo-first 𝑘 0.9735 0.9764
k = −1.499·10−3 days−1 k = −1.623𝑘·10
0.9444 =−−1.243 · 10
5 days−1
𝑑𝑎𝑦𝑠 0.7897
order = −3.677 · 10 𝑑𝑎𝑦𝑠
Zero order k0 = 1.881·10−3 days−1 0.9715 k0 = 8.921·10−3 days−1 0.9776
First-order k1 = 3.312·10−2 days−1 0.9444 k1 = 9.625·10−2 days−1 0.7897
Second-order k2 = −1.023 days−1 0.8349 k2 = −36.367 days−1 0.4789
Third order k3 = −7.431·101 days−11 0.7264 k3 = −7.288·104 days−1 0.4118

Comparing the results obtained, in the case of the TSS, the kinetic model with the
highest correlation index is the pseudo-second-order model. However, in the case of optical
density, the best correlation rate is obtained for the zero-order model. Figures 5 and 6 show
graphical representations of the adjustments that were made for the TSS and optical density
(OD) data.
The kinetic constants obtained for the fit of the optical density data show, in absolute
value, had higher values compared to those obtained for the TSS, except for the pseudo-
second-order model. This indicates that the overall growth observed is slower than the
growth of micro-organisms detected at 680 nm, including microalgae.
The best correlation rate, and therefore the best fit, was obtained for the zero-order
model when evaluating the optical density values. These results are considered satisfactory
because the optical density measurement eliminated interferences caused by bacteria or
other microorganisms in the TSS measurement.
Membranes 2022, 12, x FOR PEER REVIEW 8 of 17
Membranes 2022, 12, 982 8 of 16

Pseudo-sec- 𝐶 = 1.075 𝑔/𝐿 𝐶 = 2.316 𝑔/𝐿

0.9735 0.9764
ondThe 𝑘 growth
kinetic
order = −1.499 · 10 𝑑𝑎𝑦𝑠 can be characterised
of microalgae 𝑘 = −1.623 · 10 in𝑑𝑎𝑦𝑠
by a phase which the specific
growth rate exists
Zero order 𝑘 =until
1.881it ·reaches
10 𝑑𝑎𝑦𝑠a maximum value, 𝑘where
0.9715 it is ·maintained
= 8.921 10 𝑑𝑎𝑦𝑠 for a0.9776
specific
time. This kinetic growth
First-order 𝑘 = 3.312 · 10 𝑑𝑎𝑦𝑠model corresponds to
0.9444the one described
𝑘 = 9.625 · 10 𝑑𝑎𝑦𝑠by Monod, one of the
0.7897
most widely used [7,36]. No substrate-dependent data are available to fit this model, as
Second-order 𝑘 = −1.023 𝑑𝑎𝑦𝑠 0.8349 𝑘 = −36.367 𝑑𝑎𝑦𝑠 0.4789
the photobioreactor is assumed to operate under perfect mixing conditions. Therefore,
Third order 𝑘 = −7.431 · 10 𝑑𝑎𝑦𝑠 0.7264 𝑘 = −7.288 · 10
the nutrient concentrations (the substrate) are approximately constant throughout the𝑑𝑎𝑦𝑠 0.4118
experiment. For this reason, the fit of the experimental data to the Monod growth kinetic
model Comparing
could not be thedetermined.
results obtained, in the case of the TSS, the kinetic model with the
highest
No correlation index ismicroalgae
studies evaluating the pseudo-second-order model. However,
growth using polynomial in the case
kinetic models haveofbeen
op-
tical
found density, the best correlation
in the literature. However,rate is have
they obtained
beenfor the zero-order
evaluated model.the
to represent Figure 5 and
growth of
Figure 6 show
microalgae graphical
attached representations
to support of the adjustments that were made for the TSS
materials [37–39].
and optical density (OD) data.
0.00 0.20
-0.50
-1.00 0.15
-1.50
ln(TSS (g/L))

TSS (g/L)
-2.00
0.10
-2.50
-3.00
-3.50 0.05
-4.00
-4.50 0.00
0 20 40 60 80 0 20 40 60 80
Experimental time (days) Experimental time (days)
(a) (b)
0.20 0.00
-0.50
0.15 -1.00
ln(TSS (g/L))

-1.50
TSS (g/L)

-2.00
0.10
-2.50
-3.00
0.05 -3.50
-4.00
0.00 -4.50
0 20 40 60 80 0 20 40 60 80
Experimental time (days) Experimental time (days)
(c) (d)
70 5000
60 4000
50
3000
40
1/TSS (g/L)^2
1/TSS (g/L)

30 2000
20 1000
10 0
0
-10 -1000
-20 -2000
-30 -3000
0 20 40 60 80 0 20 40 60 80
Experimental time (days) Experimental time (days)
(e) (f)
Figure 5. Fitting data for TSS to each kinetic model assessed. (a) Pseudo-first order. (b) Pseudo-second order. (c) Zero
Figure 5. Fitting data for TSS to each kinetic model assessed. (a) Pseudo-first order. (b) Pseudo-second
order. (d) First-order. (e) Second-order. (f) Third order.
order. (c) Zero order. (d) First-order. (e) Second-order. (f) Third order.
Membranes 2022, 12,
Membranes 2022, 12, 982
x FOR PEER REVIEW 99 of
of 16
17

4 0.8
2 0.7
0.6
0
0.5
-2
ln(OD)

0.4

OD
-4
0.3
-6 0.2
-8 0.1
-10 0.0
0 20 40 60 80 0 20 40 60 80
Experimental time (days) Experimental time (days)
(a) (b)
0.8 1
0.7 0
0.6 -1
-2
0.5
-3
OD

0.4
ln(OD)
-4
0.3
-5
0.2 -6
0.1 -7
0 -8
0 20 40 60 80 0 20 40 60 80
Experimental time (days) Experimental time (days)
(c) (d)
2500 5.0E+6
2000 4.0E+6
1500 3.0E+6
1000 2.0E+6
1/OD^2
1/OD

500 1.0E+6
0 0.0E+0
-500 -1.0E+6
-1000 -2.0E+6
-1500 -3.0E+6
0 20 40 60 80 0 20 40 60 80
Experimental time (days) Experimental time (days)
(e) (f)
Figure 6. Fitting data for optical density (OD) (absorbance at 680 nm) to each kinetic model assessed. (a) Pseudo-first order.
Figure 6. Fitting data for optical density (OD) (absorbance at 680 nm) to each kinetic model assessed.
(b) Pseudo-second-order. (c) Zero order. (d) First-order. (e) Second-order. (f) Third order.
(a) Pseudo-first order. (b) Pseudo-second-order. (c) Zero order. (d) First-order. (e) Second-order.
(f) Third order.
The kinetic constants obtained for the fit of the optical density data show, in absolute
value, had higher
3.2. Membrane values compared to those obtained for the TSS, except for the pseudo-
Operation
second-order
3.2.1. Membrane model. This indicates that the overall growth observed is slower than the
Characterisation
growth of micro-organisms detected at 680 nm, including microalgae.
As described above, before starting the study in the photobioreactor, the resistance of
The best correlation rate, and therefore the best fit, was obtained for the zero-order
the cleaned membrane at different temperatures was characterised.
model when evaluating the optical density values. These results are considered satisfac-
As expected, as the temperature increases, the membrane resistance decreases, taking
tory because the optical density measurement eliminated interferences caused by bacteria
values between 6.21 × 10−3 and 5.8 × 10−3 bar·m2 ·h/L (Table 3). As previously studied by
or other
other microorganisms
authors, increasing in
thethe TSS measurement.
temperature increases the permeability of PVDF ultrafiltra-
tion membranes, or in other words, thecan
The kinetic growth of microalgae be characterised
membrane bydecreases,
resistance a phase inaswhich
shownthe
inspe-
our
cific growth rate
results [35,40]. exists until it reaches a maximum value, where it is maintained for a
specific time. This kinetic growth model corresponds to the one described by Monod, one
of the most widely used [7,36]. No substrate-dependent data are available to fit this model,
as the photobioreactor is assumed to operate under perfect mixing conditions. Therefore,
Membranes 2022, 12, 982 10 of 16

Table 3. Intrinsic resistance values of the ultrafiltration membrane at different temperatures.

Temperature (◦ C) RM 0 Values (bar·m2 ·h/L) Correlation Rate R2

20 6.21 × 10−3 0.9997
25 5.90 × 10−3 0.9994
30 5.80 × 10−3 0.9979

Once the photobioreactor study was completed after 80 days of filtration, the mem-
brane was characterised to assess its fouling. The data obtained are shown in Table 4,
with the total membrane resistances (Ri 0 ) and the fouling resistances (RF 0 ) for each of the
temperatures tested. For this purpose, the permeate flux and transmembrane pressure
results were processed to obtain the membrane permeability (k) in each case. Knowing
the relationship of this parameter with the membrane resistance (Equation (2)), Ri 0 was
obtained for each of the tests carried out. The RM 0 was obtained in the same way using the
data of the clean membrane. Subsequently, and taking into account Equation (3), the RF 0
was obtained in each case. The correlation rate corresponds to the data fit by forcing the
line through the origin.

Table 4. Total resistance and fouling resistance of the membrane at different temperatures after fouling.

Ri 0 Values RF 0 Values
Temperature (◦ C) Correlation Rate R2
(bar·m2 ·h/L) (bar·m2 ·h/L)
20 7.88 × 10−3 1.67 × 10−3 0.9997
25 7.18 × 10−3 1.28 × 10−3 0.9989
30 6.80 × 10−3 1.01 × 10−3 0.9999

The RF 0 values should be approximately the same for all temperatures, since the
fouling that has occurred on the membrane is the same for all three tests. These variations
are due to the small membrane washout that has occurred over time during the three
characterisation tests, as well as the effect of temperature, because it affects the viscosity of
the filtered fluid.
As in the case of intrinsic resistance, the total resistance of the membrane decreases as
the temperature increases. The fouling resistance values obtained were between 1.67 × 10−3
and 1.01 × 10−3 bar·m2 ·h/L; the membrane resistance increased due to fouling by approx-
imately 22%. Therefore, the fouling produced by the contents of the photobioreactor
(including microalgae) is small, as has been found in previous studies [27].
Low fouling ensures that the lifetime of the ultrafiltration membrane will be longer
because the permeability of the membrane is not significantly affected and therefore the
costs associated with pumping do not increase [41].

3.2.2. Evaluation of the Effectiveness of the Cleaning Protocol and Recovery of

Membrane Characteristics
To recover the filtering characteristics of the membrane, the cleaning protocol described
above was carried out. After each cleaning step, the membrane was characterised in water
to calculate the efficiency of each cleaning step (Table 5) according to Equation (4).
In all cases, the initial resistance of the membrane was recovered, and the overall
effectiveness of the cleaning protocol was 100%. As can be seen from the results, in the first
cleaning step, a recovery of approximately 42% was obtained. On the other hand, in the
second cleaning step, the remaining 58% was recovered, reaching the initial RM 0 values.
Both steps were effective and therefore necessary for complete cleaning of the fouled
membrane in the photobioreactor characteristics. In the tests carried out at different
temperatures, similar results were obtained, with variation in the values of the membrane
resistance because of temperature. Therefore, the optimal temperature for the cleaning
protocol is considered to be 30 ◦ C, where a lower RM 0 value is obtained.
Membranes 2022, 12, 982 11 of 16

Table 5. Results obtained for the different phases of the cleaning protocol at each of the set temperatures.

Temperature Ri 0 Values Correlation

Cleaning Step Efficiency (%)
(◦ C) (bar·m2 ·h/L) Rate R2
20 7.12 × 10−3 45.13% 0.9997
1 25 6.64 × 10−3 42.13% 0.9963
30 6.38 × 10−3 42.01% 0.9995
20 6.21 × 10−3 54.87% 0.9997
2 25 5.90 × 10−3 57.87% 0.9994
30 5.80 × 10−3 57.99% 0.9979

This protocol would be cost-effective at an industrial level because it is a discontinuous

cleaning process which can be sized according to the membranes in large installations to
minimise costs, limiting and spacing the cleaning to four times per year. This also highlights
the usefulness of this cleaning protocol, which can be used on a small scale with very good
results. Initially, only organic cleaning with sodium hypochlorite could be carried out,
minimising costs, since from the trials it is expected to be at least 50% effective. Previous
authors applied another cleaning protocol after the use of the membranes in a microalgae
culture. Their protocol was less effective than the one applied in our study because the
inorganic fouling was not successfully removed, so they suggested the use of citric acid to
complete the cleaning, and this has been done in our study [27].

3.2.3. Analysis of the Operational TMP

To determine the optimal working TMP for our study system, fluxes (J) at different
TMPs were analysed to fit the data to the Equation (5) model. In no case was it possible
to fit the proposed equation, because the studied TMPs allowed by the filtration device
were in the pressure-controlled zone, without reaching the values of the zone controlled by
matter transfer. Therefore, it was decided to fit these data to the linear part of the equation.
Figure 7 shows the data obtained for the three temperatures studied with the mem-
brane clean before starting the study cycle and after finishing it (with the membrane soiled).
From these representations, the data shown in Table 6 were obtained:

Table 6. Clean and soiled membrane resistances at different temperatures.

Clean Membrane Soiled Membrane

Temperature
RM 0 Values Correlation Ri 0 Values Correlation
(◦ C)
(bar·m2 ·h/L) Rate R2 (bar·m2 ·h/L) Rate R2
20 6.25 × 10−3 0.9985 8.02 × 10−3 0.9994
25 6.24 × 10−3 0.9994 7.26 × 10−3 0.9995
30 5.87 × 10−3 0.9993 6.68 × 10−3 0.9996

The fouling that occurs is so small that the RM 0 values obtained with water and with
the photobioreactor contents are similar both at the beginning and at the end of the cycle.
The low fouling obtained in the membrane may be due to the consortium of bacteria and
algae that exist in the MPBR, decreasing the organic load present in the medium and
therefore decreasing fouling [42]. Again, the behaviour is similar in the three studies at
different temperatures, obtaining a variation only due to the influence of temperature.
Membrane resistance increases between 28 and 13% with temperature.
Again, the resistance values obtained for the fouled membrane should be approxi-
mately the same for all temperatures, as the fouling that has occurred on the membrane
is the same for all three tests. These variations are due to the small amount of membrane
washout that has occurred over time during the three characterisation tests. As before,
these differences are observed because of the effect of temperature on the viscosity of the
filtered fluid.
Membranes2022,
Membranes 2022,12,
12,982
x FOR PEER REVIEW 12
12 of 17
of 16

50 50
45 45
40 40
35 35
30 30

J (L/m²h)
J (L/m²h)

25 25
20 20
15 y = 159.95x 15
y = 124.75x
10 R² = 0.9985 10 R² = 0.9994
5 5
0 0
0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.00 0.10 0.20 0.30
TMP (bar) TMP (bar)
(a) (b)

50 50
45 45
40 40
35 35
30 30
J (L/m²h)

J (L/m²h)
25 25
20 20
15 15
10 y = 160.17x 10 y = 137.74x
R² = 0.9994 R² = 0.9995
5 5
0 0
0.00 0.05 0.10 0.15 0.20 0.25 0.30 0.00 0.10 0.20 0.30
TMP (bar) TMP (bar)

(c) (d)
50 50
45 45
40 40
35 35
30 30
J (L/m²h)

J (L/m²h)

25 25
20 20
15 15
y = 149.72x
10 y = 170.26x 10 R² = 0.9996
5 R² = 0.9993 5
0 0
0.00 0.10 0.20 0.30 0.00 0.10 0.20 0.30
TMP (bar) TMP (bar)
(e) (f)
Figure 7. Fitting the permeate flux7.(J)Fitting
Figure and TMPthe data at different
permeate flux (J)temperatures
and TMP datato at
thedifferent
linear part of Table 20.to°C.
temperatures the(b) Soiled
linear part of
membrane at 20 °C. (c) Clean membrane at 25 °C. (d) Soiled membrane at 25 °C.◦(e) Clean membrane at 30 °C. (f) ◦ Soiled
the per-formance model. (a) Clean membrane at 20 C. (b) Soiled membrane at 20 C. (c) Clean
membrane at 30 °C.
mem-brane at 25 ◦ C. (d) Soiled membrane at 25 ◦ C. (e) Clean membrane at 30 ◦ C. (f) Soiled membrane
at 30 ◦ C.
The fouling that occurs is so small that the RM’ values obtained with water and with
the photobioreactor
As can be seen,contents are similar
the selected workingboth at the
TMP, beginning
which is 0.01and
bar,atisthe
in end of the cycle.
the linear zone
and therefore within the recommended range for the operation of this system, beingand
The low fouling obtained in the membrane may be due to the consortium of bacteria in
algae
the that exist in the MPBR,
pressure-controlled zone.decreasing
This TMPthe organicto
is similar load
thatpresent
used ininother
the medium
studiesand there-
in which
fore decreasing fouling
photobioreactors [42]. Again,are
with microalgae the used
behaviour
[20]. is similar in
Working the three
under thesestudies at different
TMP conditions
temperatures,
favours obtaining
less fouling a variation
of the membrane. only dueeffect
This to theisinfluence
explainedofby temperature.
the fact thatMembrane
at higher
resistance
TMP increases
values, between 28resistance
greater membrane and 13% iswith temperature.
caused by the compression of the layer of cells
Again,
deposited onthe resistance values obtained for the fouled membrane should be approxi-
it [43].
mately the same for all temperatures, as the fouling that has occurred on the membrane is
the same for all three tests. These variations are due to the small amount of membrane
Membranes 2022, 12, 982 13 of 16

3.3. Effluent Quality

The quality of the effluent from the photobioreactor was assessed by analysing the
TSS, turbidity, and quantification of nitrogen and phosphorus. Table 7 shows the average
values of temperature, pH, conductivity, TSS, and the turbidities of influent and effluent.

Table 7. Average values obtained for the parameters analysed in the influent and effluent.

Sample pH Temperature (◦ C) Conductivity (µS/cm)

Influent 7.71 ± 0.26 15.30 ± 0.96 1153.90 ± 95.71
Effluent 7.41 ± 0.57 14.97 ± 0.80 970.36 ± 67.10

According to Regulation (EU) 2020/741 of 25 May 2020, which establishes minimum

requirements for water reuse in agricultural irrigation, the effluent from the photobioreactor
can be reused as reclaimed water quality class A, B, C, and D [44]. In other words, it
complies with all the minimum requirements set out in that regulation.
According to Royal Decree 1620/2007, of 7 December 2007, which establishes the
legal regime for the reuse of treated water and in relation to the results obtained for the
concentration of TSS and turbidity, the effluent from our photobioreactor could be reused
for urban uses of quality 1.1 and 1.2, agricultural uses of quality 2.1, 2.2, and 2.3, industrial
uses of quality 3.1, recreational uses of quality 4.1 and 4.2, and environmental uses of quality
5.1, 5.2, and 5.3 [45]. Therefore, the effluent obtained satisfies the requirements established
for the reuse of treated wastewater in all of the uses regulated in this Royal Decree, except
for quality 3.2 (industrial uses in cooling towers and evaporative condensers).

Nitrogen and Phosphorus Removal

Table 8 shows the results obtained in the nutrient analysis: the average phosphorus
and nitrogen in the influent and effluent, the removal yields, and the consumption of
each nutrient. These last two parameters were calculated considering that the HRT in the
photobioreactor is 2 days.

Table 8. Results of nutrient analysis.

Sample Phosphorus Nitrogen

Influent 2.01 ± 1.15 ppm 36.54 ± 13.93 ppm
Removal yields 64.27 ± 0.29% 56.30 ± 0.13%
Nutrient consumption 1.09 ± 0.66 ppm 20.98 ± 9.54 ppm

The total nitrogen removal performance was slightly lower than that obtained by other
authors [7]. This effect is probably due to the presence of autotrophic microorganisms in
the wastewater. These consume the ammonium (reducing it in the photobioreactor) but
generate nitrates, increasing their concentration [20]. However, the total nitrogen removal
rate in the photobioreactor is higher than that obtained by other authors [46].
Other authors report that working times at high SRTs influence the feeding preference
of microalgae, observing that for long SRTs ammonium is preferred to nitrate, raising
nitrate levels in the effluent [28]. The preference for nitrate or ammonium will depend on
the species of microalgae grown in the photobioreactor and its operational conditions [7].
One of the possible solutions to improve the nutrient yield in the photobioreactor
would be to increase the HRT. In this way, the dilution of the culture is not so great, and
therefore, the renewal of the substrate is not so fast, thus improving the elimination yields,
as the microalgae have more time to assimilate the nutrients [12].
On the other hand, it has been observed that phosphorus concentrations have been
quite low in the influent of the photobioreactor. This event has probably conditioned
the growth of the microalgae, acting as a limiting nutrient for the culture. To achieve
higher nutrient removal yields under the same operating conditions, a higher amount of
Membranes 2022, 12, 982 14 of 16

biomass would be necessary, and this would require a higher concentration of phosphorus
in the medium. Despite this, the average removal efficiency obtained for phosphorus is
higher than that obtained in other studies [25]. Low phosphate levels cannot be ignored
as this effect can reduce nitrate uptake, limiting microalgae growth and nutrient removal
performance in the photobioreactor [7,47].

4. Conclusions
The following conclusions were reached after 80 days under continuous operation
(two days of HRT) of a microalgae membrane photobioreactor for the treatment of real
urban wastewater effluent by culturing the microalgae present in it.
On one hand, by analyzing the results obtained, it was observed that optical density
was the parameter that best described the growth kinetics of microalgae present in the
photobioreactor. The zero-order model had the highest correlation rate of 0.9776, and a
value of 8.921 × 10−3 days−1 was obtained for the kinetic constant k.
On the other hand, with respect to the membrane, there was an increase in the total
membrane resistance of approximately 22%. Therefore, under the operative conditions
tested, the fouling was low. A two-phase cleaning protocol (inorganic and organic cleaning)
was proposed to recover membrane permeability. The recoveries were approximately 42
and 58% in the inorganic and organic phases, respectively, for the temperatures tested
(20, 25, and 30 ◦ C), but RM 0 values were lower as the temperature increased, as would be
expected. At all temperatures, the overall effectiveness of the cleaning protocol was 100%.
However, the optimum temperature tested for the cleaning protocol is considered to be
30 ◦ C, where a lower RM 0 value is obtained.
Regarding the effluent obtained and the removal of the nutrients present in the treated
wastewater, total nitrogen and phosphorus removals in the algae membrane photobioreac-
tor were 56.3 and 64.27%, respectively. Therefore, taking into account other water quality
parameters, the effluent of the microalgae membrane photobioreactor could be reused for
agricultural irrigation, according to Regulation (EU) 2020/741 of 25 May 2020.
Considering the above, a microalgae membrane photobioreactor could be an appro-
priate technology for urban wastewater regeneration to be included in a conventional
activated sludge treatment plant.

Author Contributions: Conceptualization, M.d.M.M. and J.M.-P.; methodology, J.M.P.C.; formal

analysis, M.C.A.; investigation, V.D. and L.A.; writing—original draft preparation, V.D.; writing—
review and editing, J.M.P.C. and M.C.A.; supervision, project administration and funding acquisition,
J.M.-P. and M.d.M.M. All authors have read and agreed to the published version of the manuscript.
Funding: This research was funded by the Junta de Andalucía and the European Regional Develop-
ment Fund (ERDF), grant number P18-TP-4732.
Institutional Review Board Statement: Not applicable.
Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Conflicts of Interest: The authors declare no conflict of interest. The funders had no role in the design
of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or
in the decision to publish the results.

References
1. Hussian, A.E.M. The Role of Microalgae in Renewable Energy Production: Challenges and Opportunities. In Marine Ecology-Biotic
and Abiotic Interactions; IntechOpen: London, UK, 2018. [CrossRef]
2. Koller, M.; Muhr, A.; Braunegg, G. Microalgae as versatile cellular factories for valued products. Algal Res. 2014, 6, 52–63.
[CrossRef]
3. Lutzu, G.A.; Ciurli, A.; Chiellini, C.; Di Caprio, F.; Concas, A.; Dunford, N.T. Latest developments in wastewater treatment and
biopolymer production by microalgae. J. Environ. Chem. Eng. 2020, 9, 104926. [CrossRef]
Membranes 2022, 12, 982 15 of 16

4. Chia, S.R.; Chew, K.W.; Leong, H.Y.; Ho, S.-H.; Munawaroh, H.S.H.; Show, P.L. CO2 mitigation and phycoremediation of
industrial flue gas and wastewater via microalgae-bacteria consortium: Possibilities and challenges. Chem. Eng. J. 2021,
425, 131436. [CrossRef]
5. Iglina, T.; Iglin, P.; Pashchenko, D. Industrial CO2 Capture by Algae: A Review and Recent Advances. Sustainability 2022, 14, 3801.
[CrossRef]
6. Li, K.; Liu, Q.; Fang, F.; Luo, R.; Lu, Q.; Zhou, W.; Huo, S.; Cheng, P.; Liu, J.; Addy, M.; et al. Microalgae-based wastewater
treatment for nutrients recovery: A review. Bioresour. Technol. 2019, 291, 121934. [CrossRef] [PubMed]
7. Sousa, C.A.; Sousa, H.; Vale, F.; Simões, M. Microalgae-based bioremediation of wastewaters—Influencing parameters and
mathematical growth modelling. Chem. Eng. J. 2021, 425, 131412. [CrossRef]
8. Gao, F.; Peng, Y.-Y.; Li, C.; Cui, W.; Yang, Z.-H.; Zeng, G.-M. Coupled nutrient removal from secondary effluent and algal biomass
production in membrane photobioreactor (MPBR): Effect of HRT and long-term operation. Chem. Eng. J. 2018, 335, 169–175.
[CrossRef]
9. Aron, N.S.M.; Khoo, K.S.; Chew, K.W.; Veeramuthu, A.; Chang, J.-S.; Show, P.L. Microalgae cultivation in wastewater and potential
processing strategies using solvent and membrane separation technologies. J. Water Process Eng. 2021, 39, 101701. [CrossRef]
10. Tang, D.Y.Y.; Khoo, K.S.; Chew, K.W.; Tao, Y.; Ho, S.-H.; Show, P.L. Potential utilization of bioproducts from microalgae for the
quality enhancement of natural products. Bioresour. Technol. 2020, 304, 122997. [CrossRef]
11. Emparan, Q.; Jye, Y.S.; Danquah, M.K.; Harun, R. Cultivation of Nannochloropsis sp. microalgae in palm oil mill effluent (POME)
media for phycoremediation and biomass production: Effect of microalgae cells with and without beads. J. Water Process Eng.
2020, 33, 101043. [CrossRef]
12. Marbelia, L.; Bilad, M.R.; Passaris, I.; Discart, V.; Vandamme, D.; Beuckels, A.; Muylaert, K.; Vankelecom, I.F. Membrane
photobioreactors for integrated microalgae cultivation and nutrient remediation of membrane bioreactors effluent. Bioresour.
Technol. 2014, 163, 228–235. [CrossRef] [PubMed]
13. Romero-Villegas, G.I.; Burboa-Charis, V.A.; Navarro-López, E.; Cerón-García, M.C.; Acién-Fernandez, F.G.; Estrada-Alvarado,
M.I.; Rout, N.P.; Cira-Chávez, L.A. Biomass production and urban centrate nutrient removal using native microalgae tolerant to
high nitrogen concentration and temperature. J. Appl. Phycol. 2021, 33, 2921–2931. [CrossRef]
14. Ruiz, J.; Olivieri, G.; de Vree, J.; Bosma, R.; Willems, P.; Reith, J.H.; Eppink, M.H.M.; Kleinegris, D.M.M.; Wijffels, R.H.; Barbosa,
M.J. Towards industrial products from microalgae. Energy Environ. Sci. 2016, 9, 3036–3043. [CrossRef]
15. Gouveia, L.; Graça, S.; Sousa, C.; Ambrosano, L.; Ribeiro, B.; Botrel, E.P.; Neto, P.C.; Ferreira, A.F.; Silva, C.M. Microalgae biomass
production using wastewater: Treatment and costs. Algal Res. 2016, 16, 167–176. [CrossRef]
16. Abdel-Raouf, N.; Al-Homaidan, A.A.; Ibraheem, I.B.M. Microalgae and wastewater treatment. Saudi J. Biol. Sci. 2012, 19, 257–275.
[CrossRef] [PubMed]
17. Barsanti, L.; Gualtieri, P. Algae: Anatomy, Biochemistry, and Biotechnology; CRC Press: Boca Raton, FL, USA, 2005.
18. Foladori, P.; Petrini, S.; Andreottola, G. Evolution of real municipal wastewater treatment in photobioreactors and microalgae-
bacteria consortia using real-time parameters. Chem. Eng. J. 2018, 345, 507–516. [CrossRef]
19. Boelee, N.; Temmink, H.; Janssen, M.; Buisman, C.; Wijffels, R. Balancing the organic load and light supply in symbiotic
microalgal–bacterial biofilm reactors treating synthetic municipal wastewater. Ecol. Eng. 2014, 64, 213–221. [CrossRef]
20. Yang, J.; Gou, Y.; Fang, F.; Guo, J.; Lu, L.; Zhou, Y.; Ma, H. Potential of wastewater treatment using a concentrated and suspended
algal-bacterial consortium in a photo membrane bioreactor. Chem. Eng. J. 2018, 335, 154–160. [CrossRef]
21. Posadas, E.; García-Encina, P.-A.; Soltau, A.; Domínguez, A.; Díaz, I.; Muñoz, R. Carbon and nutrient removal from centrates and
domestic wastewater using algal–bacterial biofilm bioreactors. Bioresour. Technol. 2013, 139, 50–58. [CrossRef] [PubMed]
22. Wang, Y.; Ho, S.-H.; Cheng, C.-L.; Guo, W.-Q.; Nagarajan, D.; Ren, N.-Q.; Lee, D.-J.; Chang, J.-S. Perspectives on the feasibility of
using microalgae for industrial wastewater treatment. Bioresour. Technol. 2016, 222, 485–497. [CrossRef]
23. Wu, W.; Zhang, X.; Qin, L.; Li, X.; Meng, Q.; Shen, C.; Zhang, G. Enhanced MPBR with polyvinylpyrrolidone-graphene
oxide/PVDF hollow fiber membrane for efficient ammonia nitrogen wastewater treatment and high-density Chlorella cultivation.
Chem. Eng. J. 2019, 379, 122368. [CrossRef]
24. Qin, L.; Zhang, Y.; Xu, Z.; Zhang, G. Advanced membrane bioreactors systems: New materials and hybrid process design.
Bioresour. Technol. 2018, 269, 476–488. [CrossRef] [PubMed]
25. Xu, M.; Bernards, M.; Hu, Z. Algae-facilitated chemical phosphorus removal during high-density Chlorella emersonii cultivation
in a membrane bioreactor. Bioresour. Technol. 2014, 153, 383–387. [CrossRef] [PubMed]
26. Hu, W.; Yin, J.; Deng, B.; Hu, Z. Application of nano TiO2 modified hollow fiber membranes in algal membrane bioreactors for
high-density algae cultivation and wastewater polishing. Bioresour. Technol. 2015, 193, 135–141. [CrossRef] [PubMed]
27. Bilad, M.R.; Vandamme, D.; Foubert, I.; Muylaert, K.; Vankelecom, I.F.J. Harvesting microalgal biomass using submerged
microfiltration membranes. Bioresour. Technol. 2012, 111, 343–352. [CrossRef]
28. Honda, R.; Boonnorat, J.; Chiemchaisri, C.; Chiemchaisri, W.; Yamamoto, K. Carbon dioxide capture and nutrients removal
utilizing treated sewage by concentrated microalgae cultivation in a membrane photobioreactor. Bioresour. Technol. 2012, 125,
59–64. [CrossRef]
29. Peccia, J.; Haznedaroglu, B.; Gutierrez, J.; Zimmerman, J.B. Nitrogen supply is an important driver of sustainable microalgae
biofuel production. Trends Biotechnol. 2013, 31, 134–138. [CrossRef]
Membranes 2022, 12, 982 16 of 16

30. Zhang, M.; Yao, L.; Maleki, E.; Liao, B.-Q.; Lin, H. Membrane technologies for microalgal cultivation and dewatering: Recent
progress and challenges. Algal Res. 2019, 44, 101686. [CrossRef]
31. Ahmad, I.; Abdullah, N.; Koji, I.; Yuzir, A.; Muhammad, S.E. Evolution of Photobioreactors: A Review based on Microalgal
Perspective. IOP Conf. Ser. Mater. Sci. Eng. 2021, 1142, 012004. [CrossRef]
32. Sheng, A.; Bilad, M.R.; Osman, N.; Arahman, N. Sequencing batch membrane photobioreactor for real secondary effluent
polishing using native microalgae: Process performance and full-scale projection. J. Clean. Prod. 2017, 168, 708–715. [CrossRef]
33. APHA. Standard Methods for the Examination of Water and Wastewater; American Public Health Association, American Water Works
Association, Water Environment Federation: Denver, CO, USA, 2012.
34. Salgueiro Fernández, J.L. Cultivo de microalgas en aguas residuales y aprovechamiento energético de la biomasa algal. Ph.D.
Thesis, University of Vigo, Vigo, Spain, January 2019.
35. Tao, C.; Parker, W.; Bérubé, P. Assessing the role of cold temperatures on irreversible membrane permeability of tertiary
ultrafiltration treating municipal wastewater. Sep. Purif. Technol. 2021, 278, 119556. [CrossRef]
36. Lee, E.; Jalalizadeh, M.; Zhang, Q. Growth kinetic models for microalgae cultivation: A review. Algal Res. 2015, 12, 497–512.
[CrossRef]
37. de Assis, L.R.; Calijuri, M.L.; Assemany, P.P.; Berg, E.C.; Febroni, L.V.; Bartolomeu, T.A. Evaluation of the performance of different
materials to support the attached growth of algal biomass. Algal Res. 2019, 39, 101440. [CrossRef]
38. Rosli, S.S.; Kadir, W.N.A.; Wong, C.Y.; Han, F.Y.; Lim, J.W.; Lam, M.K.; Yusup, S.; Kiatkittipong, W.; Kiatkittipong, K.; Usman, A.
Insight review of attached microalgae growth focusing on support material packed in photobioreactor for sustainable biodiesel
production and wastewater bioremediation. Renew. Sustain. Energy Rev. 2020, 134, 110306. [CrossRef]
39. Mohd-Sahib, A.-A.; Lim, J.-W.; Lam, M.K.; Uemura, Y.; Ho, C.-D.; Oh, W.-D.; Tan, W.-N. Mechanistic kinetic models describing
impact of early attachment between Chlorella vulgaris and polyurethane foam material in fluidized bed bioreactor on lipid for
biodiesel production. Algal Res. 2018, 33, 209–217. [CrossRef]
40. Pascual, J.M.; Leyva-Díaz, J.; López-López, C.; Muñío, M.; Hontoria, E.; Poyatos, J.M. Effects of temperature on the permeability
and critical flux of the membrane in a moving bed membrane bioreactor. Desalin. Water Treat. 2013, 53, 3439–3448. [CrossRef]
41. Gupta, S.; Gomaa, H.; Ray, M.B. Fouling control in a submerged membrane reactor: Aeration vs membrane oscillations. Chem.
Eng. J. 2022, 432, 134399. [CrossRef]
42. Xie, B.; Gong, W.; Yu, H.; Tang, X.; Yan, Z.; Luo, X.; Gan, Z.; Wang, T.; Li, G.; Liang, H. Immobilized microalgae for anaerobic
digestion effluent treatment in a photobioreactor-ultrafiltration system: Algal harvest and membrane fouling control. Bioresour.
Technol. 2018, 268, 139–148. [CrossRef]
43. Liu, B.; Qu, F.; Liang, H.; Van der Bruggen, B.; Cheng, X.; Yu, H.; Xu, G.; Li, G. Microcystis aeruginosa -laden surface water
treatment using ultrafiltration: Membrane fouling, cell integrity and extracellular organic matter rejection. Water Res. 2017, 112,
83–92. [CrossRef]
44. European Commision. Reglamento (UE) 2020/741 del Parlamento Europeo y del Consejo, de 25 de Mayo de 2020, Relativo a Los
Requisitos Mínimos para la Reutilización del Agua; European Commision: Brussels, Belgium, 2020; Volume 2019, pp. 2–32. Available
online: https://eur-lex.europa.eu/resource.html?uri=cellar:e8951067-627c-11e8-ab9c-01aa75ed71a1.0024.02/DOC_2&format=
PDF (accessed on 31 August 2022).
45. Government of Spain. Real Decreto 1620/2007, de 7 de Diciembre, Por El Que Se Establece El Régimen Jurídico de la Reutilización
de las Aguas Depuradas. BOE 2007, 294, 50639–50661. Available online: https://www.boe.es/eli/es/rd/2007/12/07/1620
(accessed on 31 August 2022).
46. García, D.; Alcántara, C.; Blanco, S.; Pérez, R.; Bolado, S.; Muñoz, R. Enhanced carbon, nitrogen and phosphorus removal from
domestic wastewater in a novel anoxic-aerobic photobioreactor coupled with biogas upgrading. Chem. Eng. J. 2017, 313, 424–434.
[CrossRef]
47. Suthar, S.; Verma, R. Production of Chlorella vulgaris under varying nutrient and abiotic conditions: A potential microalga for
bioenergy feedstock. Process Saf. Environ. Prot. 2018, 113, 141–148. [CrossRef]