International Journal of Environmental Research

https://doi.org/10.1007/s41742-018-0092-0 (0123456789().,-volV)(0123456789().,-volV)

RESEARCH PAPER

Algal Nutrient Removal from Wastewater in Fed-Batch Operated

Photobioreactor
Hülya Beydeş1 • Ilgi Karapinar1

Received: 6 December 2017 / Revised: 20 March 2018 / Accepted: 23 April 2018

Ó University of Tehran 2018

Abstract
Photobioreactor design and determination of operation conditions for pollution removal from wastewater by algae
Chlorella vulgaris is a challenge to develop a low cost bioprocess technology alternative to conventional bacterial nutrient
removal processes. Fed-batch operation strategy, commonly used in bacterial wastewater treatment, was applied to algal
nutrient removal to determine the operating conditions and to evaluate its photobioreactor behavior. The process was
operated at different influent NH4–N (12.6–32 mg/L), PO4–P concentrations (3.2–8 mg/L) and then at varying N/P ratio of
4.6/1 and 7.3/1 at NH4–N concentration of 42 ± 2 mg/L. The other operating conditions were adjusted as light inten-
sity = 1100 lx, L/D = 20 h/4 h and hydraulic retention time, HRT = 3 days. The results indicated that nutrient removal in
fed-batch operated photobioreactor was mainly affected by the influent NH4–N concentration. The highest NH4–N and
PO4–P removal efficiencies were obtained as 84 and 83% at NH4–N = 12.6 mg/L and PO4–P = 3.2 mg/L influent con-
centrations, respectively. The best N/P ratio for efficient removal of nitrogen (75%) and phosphorus (75%) was determined
to be N/P = 4.6. Fed-batch operated photobioreactor provided excess algae growth through which nutrient uptake was
enhanced.
Graphical Abstract

30 100 8 100

25 7
80
PO4-P Concentration, mg/L
NH4-N Concentration, mg/L

80
6
Percent Removal

Percent Removal
20
60 5 60
15 4
40 40
3
10
2
20 20
5
1

0 0 0 0
0 10 20 30 40 0 2 4 6 8 10
Initial NH4-N Concentration, mg/L Initial PO4-P Concentration, mg/L

Keywords Algae  Fed-batch  Nutrient removal  Nitrogen  Phosphorus  Photobioreactor

Introduction
& Ilgi Karapinar
ilgi.karapinar@deu.edu.tr Nitrogen and phosphorus removal from wastewater are
1
Department of Environmental Engineering, Dokuz Eylül
commonly realized in bacterial nutrient removal processes
University, Buca, Izmir, Turkey (BNP). Since, the biochemical reactions, bioprocesses and

123

environmental conditions in BNP are well-defined. The
processes contain sequential combinations of anaerobic,
aerobic and anoxic zones through which nitrification,
denitrification and enhanced phosphorus removal by luxury
uptake into bacterial cell are achieved. Sequencing batch
reactor (SBR), 4-Stage Bardenpho and 5-Stage Bardenpho
with enhanced phosphorus removal, University of Cape
Town (UCT), Virginia Initiative Plant (VIP) are well-
known processes for BNP (Ekama 2011). The required
HRT for these process ranges between 7 and 15 h (Judd
et al. 2015). Relatively recent advance on this subject is the
energy efficient biological nutrient removal mechanisms
namely anaerobic ammonium oxidation (ANAMOX),
system for high rate ammonium removal over nitrite
(SHARON), completely autotrophic nitrogen removal over
nitrite (CANON) and coupled aerobic–anoxic nitrous
decomposition (CANDO) (Nancharaiah et al. 2016).
However, nutrient removal still requires cost effective,
easy operation bioprocess alternatives. It has been
demonstrated that algae has got a considerable potential to
remove nitrogen and phosphorus from wastewater, and
algal photobioreactor (PBR) could be an alternative to
BNP. The nutrient removal mechanism by algae include
uptake into the cell and stripping ammonia to the atmo-
sphere or precipitation of phosphate through elevated pH
resulted from algal metabolic activity (Aslan and Kapdan
2006; Gao et al. 2014; Munoz and Guieysse 2006; Garcia
et al. 2006). There are certain advantages of PBR over BNP
such as low operation cost, oxygenated effluent discharge
into receiving water body, utilization of assimilated nitro-
gen and phosphorus as fertilizer, thereby, lower sludge
management requirements compared to BNP and finally
nutrient removal without carbon requirement makes PBR
attractive for carbon limited but nutrient rich secondary
effluent treatment (Judd et al. 2015). The other significant
fact about algal processes is simultaneous nutrient removal
and biofuel production, such as biodiesel, through the
growth of algae or use of biomass as substrate for biohy-
drogen (Tao et al. 2017; De Alva et al. 2013; Yang et al.
2011; Ji et al. 2015). The most commonly used algae
cultures for nutrient removal are Chlorella sp. (Wang et al.
2010; Yang et al. 2011; Liang et al. 2013; Lu et al. 2015;
Tao et al. 2017), Spirulina (Chang et al. 2013) and Sce-
nedesmus sp. (Ruiz-Marin et al. 2010; Li et al. 2010; De
Alva et al. 2013; Prandini et al. 2016). Nutrient removal
with other algae cultures as Ourococcus multisporus (Ji
et al. 2013), Galdieria sulphuraria (Henkanatte-Gedera
et al. 2017), Klebsormidium sp. and Stigeoclonium sp. (Liu
et al. 2016), Coelastrella sp. (Luo et al. 2016) and Des-
modesmus sp. (Ji et al. 2013) have been also investigated. It
has been revealed that removal of nutrients could vary
depending on the culture used.
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International Journal of Environmental Research
Photobioreactor design and operation are the key factors
to obtain high yield of biomass and nutrient removal. The
nutrient removal performances of the cultures vary
depending on operation conditions such as hydraulic
retention time (HRT), N/P ratio in the feed, light regime,
light intensity etc. Various algae photobioreactor (PBR)
have been used for nutrient removal from synthetic or real
wastewater (Judd et al. 2015). The well known suspended
algae wastewater treatment bioprocesses are High Rate
Algae Ponds (HRAP) (Sutherland et al. 2014), corrugated
raceways (Olguı́n et al. 2003), tubular photobioreactor
(Cattaneo et al. 2003), sequencing batch system (Huang
et al. 2015), airlift photobioreactor (Tao et al. 2017). Algae
harvesting problems encountered in suspended growth
bioprocess and high algae biomass requirement for either
efficient removal or for biomass productivity need devoted
attentions to immobilized photobioreactors. Immobiliza-
tion of algae in/on sodium-alginate beads is one of the most
common methods (Ruiz-Marin et al. 2010). Alternatively,
immobilization on solid surfaces has been applied. Such as
submerged tube biofilm photobioreactor (Kapdan and
Aslan 2008), fed-batch operated rotating algal biofilm
reactor (Shayan et al. 2016), air lift reactor containing solid
carries (Tao et al. 2017), algae based membrane reactors
(Xu et al. 2014; Tang and Hu 2016; Gao et al. 2016) have
been used for high-density algae cultivation and nutrient
removal. Judd et al. (2015) has reviewed recent research
results about the algal photobioreactors and nutrient abet-
ment to present the state of the art for PBRs. It was
emphasized that required HRT for PBR is between 2 and
5 days irrespective of technology configuration, but it is in
the range of 7–15 h in BNR. Long HRT requirement in
algal process makes it unable to compete with BNP. Since,
algal nutrient removal is attained through mainly uptake of
nutrient into cell, biomass concentration should be kept
high enough to achieve high uptake which provide better
nutrient removal with shortening the HRT. It is evident that
high biomass concentration can be achieved by immobi-
lized PBR, homogeneous environment for substrate and
light availability are advantageous of suspended growth
algae photobioreactors. Fed-batch operation is one of the
bioprocess technologies used for wastewater treatment.
The process provides high biomass concentration, easy
control of environmental conditions, resistance to shock
substrate, toxic or inhibitory compound loadings. By con-
sidering the advantages of fed-batch cultivation about high
biomass and tolerance to high substrate concentrations, the
study was designed to investigate the performance of fed-
batch operated photobioreactor for nutrient removal by
algae. It was aimed to determine the effect of some oper-
ating parameters as influent nutrient concentrations and
N/P ratio on algae growth, Chla synthesis, nitrogen and
phosphorus removal at HRT = 3 days.
International Journal of Environmental Research
Materials and Methods
Microorganism
The green algae Chlorella vulgaris was obtained from Ege
University, Bioengineering Department, Izmir, Turkey in
pure form and then cultivated in Dokuz Eylül University,
Department of Environmental Engineering, Izmir, under
sterile conditions.
Culture Medium
Synthetic wastewater was made up of MgSO47H2O,
1000 mg/L CaCl2, 84 mg/L, and 0.5 mL/L trace elements
solution. Nitrogen and phosphorus sources were NH4Cl
and KH2PO4, respectively. The trace elements solution was
composed of H3BO3, 57 mg/L FeSO47H2O, 25 mg/L;
ZnSO47H2O, 44 mg/L; MnCl24H2O, 7 mg/L; MoO3,
35 mg/L; CuSO45H2O, 8 mg/L; Cu(NO3)26H2O, 2.5 mg/
L; Na2EDTA, 250 mg/L; and NaHCO3, 2500 mg/L.
Experimental Studies
A cylindrical glass photobioreactor with total volume of
5 L was used in the study. The temperature of liquid media
was kept constant at 26 ± 1 °C by using aquarium heaters.
pH was controlled at pH 6.5–7.0 with the addition of 5%
KOH and 10% CH3COOH when required. Light was pro-
vided with 36 W fluorescent lamps in reciprocal sides of
the photobioreactor. The process was started with initial
liquid volume of V0 = 500 mL containing dense culture of
Chlorella vulgaris (X0 = 700 ± 50 mg/L) at the beginning
of each operation and then, fed with synthetic wastewater
at constant flow rate (Q = 1.1 ± 0.1 L/day) by a peristaltic
pump to obtain HRT = 3 days. The liquid reaction volume
was 4.6 ± 0.1 L at the end of the operation. As the reac-
tion completed after 3 days of operation, the biomass set-
tled to 500 mL influent volume, the supernatant was
withdrawn by using a peristaltic pump within 1 h. Biomass
concentration in the settled sludge was measured and the
excess biomass was withdrawn to keep the influent con-
centration around X0 = 700 ± 50 mg/L. The reactor was
aerated to provide CO2 and mixing.
Influent NH4–N concentration was varied between
NH4–N0 = 12.6–32 mg/L. Light intensity inside of the
reactor L/D cycle and N/P ratio were 1100 lx, 20 h/4 h and
4/1, respectively, in the first part of the fed-batch experi-
ments. In the second part, N/P ratio was varied between
4.6/1 and 7.3/1 at constant NH4–N0 = 42 ± 2 mg/L. The
other parameters were adjusted as light intensity = 1100 lx
at the inside of the reactor with L/D = 20 h/4 h.
Analytical Methods
Nitrogen and Phosphorus Analysis
A daily sample withdrawn from the reactor was centrifuged
at 5000–6000 rpm for 15 min to separate algae. The con-
centration of ammonia nitrogen and phosphorus were
measured in clear supernatant by colorimetric methods by
using Merck kits for ammonium and phosphorus analysis
with the kit number 1.14752 and 1.14842, respectively.
Biomass Concentration and Chlorophyll a Analyses
Initial and final algae biomass concentration were deter-
mined by filtration of mixed liquid on pre-weighted and
dried filter paper (Whatman GF/C) according to SM2540-D
(APHA SM-2005).
Chla analysis was conducted on algae biomass obtained
from centrifugation of 10 mL of algal suspension at 5000–
6000 rpm for 30 min. The cells were re-suspended in 3 mL
methanol and boiled for about 5 min in a water bath. The
samples were cooled to room temperature, the volume was
made up to 5 mL by adding distilled water and then the
absorption (A) of the pigment extract in a spectropho-
tometer at the given wavelength against a solvent blank
was determined. The chlorophyll a concentration in the
extract was calculated by using the Eq. 1 (Becker 1994):
Chlorophyll a ¼ ð16:5  A665Þ  ð8:3  A650Þ: ð1Þ
Results and Discussions
Transition Behavior of Fed-Batch Operated
Photobioreactor
The transition behavior of the system was investigated at
influent NH4–N concentration of 22.4 mg/L and PO4–P
concentration of 5.6 mg/L. The concentrations of these
nutrients as well as biomass and Chla were monitored
throughout the operation period. There is substantial dilu-
tion of substrate or pollutant in the fed-batch operation
process. In order to show that dilution was not the main
mechanism that determines the effluent quality, theoretical
concentrations of all monitored parameters were calculated
by considering the dilution and presented in the figures.
Figure 1a indicates the variation of effluent and theoretical
NH4–N concentrations and removal efficiency with time.
The concentration of nitrogen in the system did not change
substantially after second day of operation. The concen-
tration was 10.4 mg/L on day 2 and decreased to 8.8 mg/L
on day 3 with 55% removal efficiency regarding to the
calculated nitrogen concentration (19.7 mg/L). Since the
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variation of effluent nitrogen concentration in the system
was not substantial on third day, the process was not
operated further.
Similarly, Fig. 1b shows PO4–P removal performance of
fed-batch operated algal photobioreactor. The theoretical
concentration in the liquid phase was around 4.7 mg/L on
day 2 and day 3. The effluent concentration did not change
for the operation periods between 2 and 3 and the final
PO4–P was obtained as 2.1 mg/L with 56% removal effi-
ciency with respect to the theoretical concentration.
Biomass and Chla concentrations were monitored to
understand if there was growth and if the culture was
active. For the case of Chla, it was 15.5 mg/L at the
beginning of the operation but decreased to 8.5 mg/L at the
end of 3 days operation period (Fig. 2a). These results
could be evaluated as that concentration decreases and no
new cells or chlorophyll was synthesized. However, if there
were no Chla production, the final concentration would be
1.9 mg/L at the end of operation with the effect of dilution
in fed-batch operation. Instead, the measured Chla con-
centration is substantially higher than this calculated value.
The algae growth showed almost the same result (Fig. 2b).
The influent biomass concentration during start up was
671 mg/L. The calculated biomass concentration at the end
of 3 days operation was 82 mg/L, but the observed one was
433 mg/L. The results about biomass and Chla concentra-
tions showed that there was a substantial growth of the
algae in the system, the culture was active and the main
nutrient removal mechanism was nutrient up-take through
growing algae.
Effect of Influent NH4–N Concentration
Figure 3 depicts the variation of nitrogen removal with the
influent NH4–N concentration which was varied between
NH4–N concentration at the end of operation increased
from 1.8 to 10.6 mg/L with the increase influent NH4–N
concentration from 12.6 to 32 mg/L. The rate of ammonia
removal also increased from 3.6 to 7.1 mg/L day. Theo-
retical final NH4–N concentration would be 11 and
28.4 mg/L corresponding to feed nitrogen concentrations.
By considering the dilution in the system, the final removal
efficiencies with the contribution of algae were obtained as
84 and 63% for the influent concentrations of 12.6 and
32 mg/L, respectively. PO4–P concentration in the feed
(PO4–P0) was varied between 3.2–8 mg/L depending on
NH4–N0 concentrations by keeping N/P ratio = 4/1
(Fig. 4). Effluent PO4–P concentration was 0.4 mg/L for
PO4–P0 = 3.2 mg/L but it increased to 2.3 mg/L for PO4–
P0 = 8 mg/L resulting in decreasing in the percent removal
from 83 to 68%, respectively. But the rate of removal
increased from 0.9 to 1.9 mg/L day.
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Variations of observed and calculated biomass concen-
trations in fed-batch operation for different nitrogen and
phosphorus concentrations were presented in Fig. 5. The
influent biomass concentration during the start up period
was around 700 ± 50 mg/L. The final biomass concen-
tration at the end of operation periods for different influent
NH4–N was between 400 and 459 mg/L. On the other
hand, the biomass concentration would be around 90 mg/L
due to dilution with the feeding. The observed biomass
concentration was substantially higher compared to the
theoretical one.
Similarly, the effect of influent nutrient concentration on
Chla was evaluated and presented in Fig. 6. Fed-batch
operation was started with a dense culture of C. vulgaris as
10% of reactor volume (V0 = 500 mL). Chla content of the
biomass in start-up volume was between 11.2 and 15.5 mg/
L for different influent nutrient concentrations. However,
the final Chla concentrations decreased from 10.3 to
4.9 mg/L with the increase in influent from NH4–
N0 = 12.6 mg/L to NH4–N0 = 32 mg/L. The amount of
Chla per biomass showed almost the same trend and the
ratio decreased from 0.022 to 0.012 with the increase in the
feed nutrient concentration. The excess ammonia nitrogen
(NH4–N0 [ 22.4 mg/L) could have had adverse affect on
Chla synthesis as well as nutrient removal although there
was high growth of algae. These results depicted that C.
vulgaris is very effective in removing low influent NH4–N
concentrations (NH4–N0 = 12.6 mg/L). The performance
of the culture substantially decreases at higher concentra-
tions such as NH4–N0 = 32 mg/L. Therefore, it can be
concluded that C. vulgaris containing fed-batch system can
be operated with 12.6 mg/L influent NH4–N concentration
for efficient removal.
Effect of N/P Ratio on Nitrogen and Phosphorus
Removal
N/P ratio is another parameter that affects nutrient removal
in any biological treatment system. The limitation in one of
the nutrients directly will certainly effect the removal of
the other. Therefore, fed-batch operation was conducted at
different N/P ratios between 4.6/1 and 7.3/1. NH4–N con-
centration was kept constant around 42 ± 3 mg/L while
phosphate concentration was gradually decreased from 8.7
to 6.1 mg/L.
The effect of N/P ratio on NH4–N removal is given in
Fig. 7. Effluent NH4–N concentration decreased from 17.8
to 8.8 mg/L for the N/P = 4.6 and N/P = 7.3, respectively.
The final NH4–N concentrations would have been in the
range of 35–40 mg/L if only dilution in operation was
considered. The contribution of algae to nitrogen removal
was over 75% for N/P = 4.6 and around 55% for
N/P = 7.3. Nitrogen removal rate showed a decreasing
International Journal of Environmental Research
Fig. 1 a Variation of effluent NH4–N concentration, theoretical NH4–
N concentration and NH4–N removal efficiency with time at (NH4–
N)0 = 22.4 mg/L (filled square line: theoretical NH4–N concentra-
tion, filled triangle line: effluent NH4–N concentration, filled circle
line: NH4–N removal efficiency), b Variation of effluent PO4–P
concentration, theoretical PO4–P concentration and PO4–P removal
efficiency with time at PO4–P0 = 5.6 mg/L (filled square line:
theoretical PO4–P concentration, filled triangle line: effluent PO4–P
concentration, filled circle line: PO4–P removal efficiency)
trend from 11 to 8 mg/L day with respect to increasing in
ratio. The main reason for low nitrogen removal at
N/P = 7.3 is the phosphorus limitation in the media. Sim-
ilarly, effluent PO4–P concentrations increased from 1.4 to
3.1 mg/L with increasing N/P ratio (Fig. 8). As a result
percent removal decreased from 75 to 60%, respectively.
Phosphorus removal rate decreased from 2.4 to 1.0 mg/
L day as well. Excess nitrogen at N/P = 7.3 might have
caused substrate inhibition which resulted in low phos-
phorus removal.
Fig. 2 a Variation of theoretical Chla concentration and effluent
Chla concentration with time at (NH4–N)0 = 22.4 mg/L (filled square
line: theoretical Chla concentration, filled triangle line: effluent
Chla concentration). b Variation of biomass concentration and
effluent biomass concentration with time at (NH4–N)0 = 22.4 mg/L
(filled square line: theoretical biomass concentration, filled triangle
line: effluent biomass concentration)
Figure 9 depicts the variation of algae concentration in
fed batch operation. The influent biomass concentration
was around 700 ± 40 mg/L. The calculated biomass con-
centration with considering dilution in the system was
88 ± 6 mg/L. The observed ones were 484 mg/L for
N/P = 4.6 and 384 mg/L for N/P = 7.3. There was excess
growth of algae in comparison to calculated biomass con-
centration. The decrease in the biomass concentration can
be explained as adverse effect of phosphorus limitation at
N/P = 7.3. Fed-batch operation was started with concen-
trated biomass, the Chla concentration at the start-up phase
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Fig. 3 Variation of effluent NH4–N concentrations and removal
efficiency with influent NH4–N concentrations (filled square line:
theoretical NH4–N concentration, filled triangle line: effluent NH4–N
concentration, filled circle line: NH4–N removal efficiency)
Fig. 4 Variation of PO4–P concentrations and removal efficiency
with influent PO4–P concentrations (filled square line: theoretical
PO4–P concentration, filled triangle line: effluent PO4–P concentra-
tion, filled circle line: PO4–P removal efficiency)
was in the range of 11.2–18.7 mg/L (Fig. 10). The
observed Chla concentrations for the different N/P ratios
were between 6 and 9 mg/L, respectively.
Experimental studies indicated that, C. vulgaris can
efficiently remove NH4–N and PO4–P at low N/P (N/
P = 4.6) with a removal efficiency 75%. However, N/P (N/
P = 4.6) significantly affect biomass and Chla concentra-
tion in the system. Both Chla and biomass concentration
decreases with the increasing ratio. This result can be
explained as the effect of phosphorus on the growth and the
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International Journal of Environmental Research
Fig. 5 Effect of influent NH4–N concentration on biomass concen-
tration (filled circle line: theoretical biomass concentration, open
circle line: observed biomass concentration)
Fig. 6 Effect of influent NH4–N concentration on Chla concentration
and Chla/biomass ratio (filled circle line: Chla/biomass, open circle
line: Chla concentration)
Chla synthesis. The phosphorus concentration decreases
with the increasing ratio, so that makes phosphorus the
limiting substrate. It could act as the growth limiting sub-
strate at high N/P ratios.
The results of this study are comparable with that of
other batch and continuous algal photobioreactors. There
are numerous batch algal nutrient removal studies. Such as,
Wang et al. obtained 82 and 83% NH3–N (33 mg/L) and
PO4–P (5.66 mg/L) removal with Chlorella sp. from
domestic wastewater (Wang et al. 2010) by batch
International Journal of Environmental Research

cultivation. Yang et al. (2011) reported that Chlorella

ellipsoidea YJ1 could achieve more than 99 and 95%
removal of TN and TP from secondary effluents with TN
and TP concentrations of 16.7 and 0.08 mg/L, respectively.
C. vulgaris exhibited 29% NH4–N and 55% total phos-
phorus removal ability at 20 mg NH4–N/L and 5 mg/L TP
influent concentrations in 6 days batch cultivation (Liang
et al. 2013). Lu et al. (2015) obtained between 45 and
60.5% NH3–N removal by Chlorella sp. from excess NH3–
N containing meat processing wastewaters as 193 and
101 mg/L and complete removal or NH3–N from 2 to
8.3 mg/L NH3–N containing wastewaters.
In continuously operated bioprocesses, HRT is one of
the factors affecting the nutrient removal performance. The
effluent quality could vary depending on the suspended and
immobilized bioprocess types, influent nutrient concentra-
tions or used algal species. The reported HRT ranges since
year 2012 for immobilized and suspended algal photo- Fig. 8 Variation of effluent, theoretical PO4–P concentration and
bioreactors have been summarized by Judd et al. (2015). removal efficiency with time at N/P ratio (filled square line:
theoretical PO4–P concentration, filled triangle line: effluent PO4–P
HRT for suspended culture systems like High Rate Algal
concentration, filled circle line: percent removal)
Ponds (HRAP) is in the range of 2–10 days with 59–79% N
and 12–79% P removal efficiencies. Suspended airlift
photobioreactor was operated by Tao et al. (2017) and
dissolved inorganic nitrogen (DIN) and dissolved inorganic
phosphorus removal (DIP) beside to algal productivity and
lipid production at 10 days HRT were investigated.
Removal efficiencies were 53% DIN and 67% DIP for
influent concentrations of 15.8 and 3.07 mg/L, respec-
tively. Nutrient removal in CSTR at HRT = 4 days were
63% for TN and 95% for TP for the influent concentrations
of around 11.84 mg TN/L and 2.42 mg TP/L. Fed-batch

Fig. 9 Variation of biomass concentration and effluent biomass

concentration with time at N/P ratio (filled circle line: theoretical
biomass concentration, open circle line: observed biomass
concentration)

operated process used in this study provided higher

Fig. 7 Variation of effluent, theoretical NH4–N concentration and
removal efficiency with N/P ratio (filled square line: theoretical NH4–
N concentration, filled triangle line: effluent NH4–N concentration,
filled circle line: percent removal)
removal efficiencies in a relatively shorter HRT (3 days)
compared to other reported ones with suspended algal
culture bioprocess.
HRTs in immobilized systems such as membrane pho-
tobioreactor is lower around 1–2 days compared to sus-
pended growth ones. Xu et al. (2014) reported 66 ± 9% TP
removal in HRT = 1 day with the excess algae growth
around 4840 mg/L in an algae based membrane bioreactor.

123

Fig. 10 Variation of Chla concentration and Chla/biomass ratio with
N/P ratio (filled circle line: Chla/biomass, open circle line
Chla concentration)
TN and TP removal in another algae based membrane
reactor with HRT = 1 day was 64 and 100% for TN and
TP, respectively, for the influent concentrations of TN =
11.89 mg/L and TP = 2.41 mg/L (Tang and Hu 2016). It
was found that nitrogen removal performance of membrane
reactor was three times higher than that of completely
mixed reactor (CSTR) under the same operation condi-
tions. Gao et al. (2016) studied on operating a membrane
photobioreactor at HRT = 1 day with 86% TN and 83% TP
removal efficiencies at relatively low influent concentra-
tions of TN = 6.81 mg/L and TP = 0.41 mg/L. A novel
approach to immobilized reactor operation was reported by
Shayan et al. (2016). Rotating biological contactor was
operated fed-batch at HRT = 2 days. The resulting
removal efficiencies were 94% TN and 95% TP at rela-
tively high nutrient concentrations as around TN = 49.5
mg/L and TP = 14.95 mg/L. Tao et al. (2017) used solid
carrier containing biofilm airlift photobioreactor for DIN
and DIP removal. The process resulted in 62 and 72%
removal at DIN = 15.86 mg/L and DIP = 3.07 mg/L
influent concentrations, but, at HRT = 10 days. The results
of our study are comparable with the immobilized systems
in terms of HRT. Removal efficiencies were 84% NH4–N
and 83% PO4–P at 12.6 and 3.2 mg/L influent concentra-
tions, respectively, at HRT = 3 days.
Phosphorus and ammonia removal by algae could be
enhanced through ammonia stripping and chemical pre-
cipitation of phosphorus with the elevated pH of the liquor
due to algae photosynthesis (Gao et al. 2014; Munoz and
Guieysse 2006; Garcia et al. 2006). It was also reported
that acidic conditions could adversely affect the nutrient
removal performance of algae (Scenedesmus sp. LX1)
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International Journal of Environmental Research
when ammonium is the nitrogen source. Therefore, pH
should be kept above 7 for nutrient removal by algal
growth (Li et al. 2010). pH of the media in our study was
controlled at neutral pH to eliminate ammonia stripping or
phosphorus precipitation. Therefore, the main nutrient
removal mechanism in this study could be attributed to the
cell uptake.
Conclusions
Fed-batch operated photobioreactor with Chlorella vul-
garis provided efficient nitrogen removal and biomass
synthesis. Nutrient removal efficiencies reached up to
about 83% for both NH4–N and PO4–P at relatively short
HRT of 3 days. The sole nutrient removal mechanism in
the study was uptake of nutrient into algae cell. The excess
biomass and Chla synthesis show presence of active bio-
mass in the system. Removal of nutrients through enhanced
cell uptake makes the process more advantageous regard-
ing to nitrogen and phosphorus recycle such as to be used
as fertilizer and no phosphorus residue as chemical pre-
cipitate. The results of this study indicated that fed-batch
operated photobioreactor system for nutrient removal is a
promoting approach to obtain efficient nutrient removal
and excess biomass synthesis which could be used for other
purposes as fertilizer or third generated biomass in biofuel
production. Fed-batch operated algal nutrient removal
process could achieve higher performance at shorter HRTs
by optimizing reactor design for better light penetration
and the operating parameters such as initial biomass con-
centration, light intensity, CO2 availability.
Acknowledgements This study was financially supported by Dokuz
Eylül University Research Fund with Grant number 03.KB.FEN.069,
Izmir, Turkey. The authors thank Prof. Dr. Rengin Eltem for sup-
plying algae culture.
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