J Appl Phycol
DOI 10.1007/s10811-013-9982-x
Evaluation of microalgal consortia for treatment of primary
treated sewage effluent and biomass production
Nirmal Renuka & Anjuli Sood & Sachitra K. Ratha &
Radha Prasanna & Amrik S. Ahluwalia
Received: 26 August 2012 / Revised and accepted: 7 January 2013
# Springer Science+Business Media Dordrecht 2013
Abstract The present investigation was aimed towards an-
alyzing the potential of consortia of native filamentous
microalgal strains (MC2), native unicellular microalgal
strains (MC3), and selected microalgae from germplasm
(MC1) in terms of nutrient removal, water quality improve-
ment, and biomass production using primary treated sewage
water. Highest NO3-N (90 %) and PO4-P (97.8 %) removal
was obtained with MC2-inoculated sewage water. Highest
decrease in total dissolved solids to 806 from 1,120 mg L−1
and highest increase in dissolved oxygen of 9.0 from
0.4 mg L−1 were obtained using MC2-inoculated sewage
water on the sixth day. The biomass production was also
highest in MC2 (1.07 g L−1) followed by MC1 and MC3
(0.90 and 0.94 g L−1, respectively) on the sixth day. The
consortium of filamentous strains from native environment
not only proved promising in nutrient removal efficiency
but also led to enhanced biomass. The present study
highlighted the utility of such a consortium for sewage
wastewater treatment and the promise of sewage water as
a growth medium for biomass production.
Keywords Biomass . Consortium . Filamentous
microalgae . Nutrient removal . Sewage water
Electronic supplementary material The online version of this article
(doi:10.1007/s10811-013-9982-x) contains supplementary material,
which is available to authorized users.
N. Renuka : A. S. Ahluwalia
Department of Botany, Panjab University, Chandigarh 160014,
India
A. Sood
Department of Botany, University of Delhi, Delhi 110007, India
S. K. Ratha : R. Prasanna (*)
Division of Microbiology, Indian Agricultural Research Institute,
New Delhi 110012, India
e-mail: radhapr@gmail.com
Introduction
The increasing scarcity of water in the world, along with
rapid population increase in urban areas, and improper
dumping of wastes into aquatic bodies are areas of seri-
ous concern, which emphasize the need for suitable and
effective treatment options. According to a WHO report
(2000), only 35 % of total sewage wastewater actually
undergoes treatment to secondary level in Asia, while in
African countries, no technique is being employed for
the treatment of sewage wastewater (Singh et al. 2012).
In India, a survey of Central Pollution Control Board
stated that out of the total sewage generated from urban
areas, only 35 % is being treated, with a capacity gap of
65 % (CPCB 2009). Apart from the lack of sufficient
means to treat sewage water, another problem associated
is the presence of excess of nutrients, especially N and
P. This, in turn, leads to ecological problems such as
eutrophication, algal blooms, uncontrolled spread of cer-
tain aquatic macrophytes, oxygen depletion, and loss of
key species, and degradation of fresh water ecosystems
(Wang et al. 2010; Doria et al. 2012). Traditional treatment
systems such as aerobic and anaerobic reactors are costly,
involve complex operation, generate a great volume of waste
sludge, and require a high-energy input. Therefore, there is a
need to develop a cost-effective, eco-friendly technology that
utilizes economically viable inputs, for its acceptability at
commercial level.
Phytoremediation or the use of plants or algae for treat-
ment of wastewater is a suitable option (Sood et al. 2011).
Microalgae, a broad category comprising of eukaryotic
microalgae and cyanobacteria, has recently been exploited
for the removal of nutrients from wastewater, because of
their high requirement of N and P for their growth (Mata et
al. 2012). Wastewaters also provide a suitable medium,
supplying most of the necessary nutrients for algae growth,
thereby, significantly reducing the cost associated with

credits for wastewater treatment, as well as mitigation of the
greenhouse gas emissions (Pittman et al. 2011). Another
advantage of this system is that the microalgal biomass
generated from wastewater treatment plant can be utilized
for the production of bioenergy, food, animal feed, pharma-
ceuticals, and fertilizers (Rawat et al. 2011).
Recently, a number of reports employing mainly uni-
cellular microalgae have revealed that wastewater can be
effectively used as growth medium for the cultivation
and biomass production (Cho et al. 2011; Sydney et al.
2011; Zhou et al. 2012a, b; Renuka et al. 2013). How-
ever, a serious drawback associated with the application
of unicellular microalgae in sewage treatment is their
microscopic dimensions (0.5–30 μm), which makes bio-
mass harvesting not economically viable, besides being
a time-consuming process (Molina Grima et al. 2003).
In this context, filamentous microalgae, with dimensions
of approximately 200 μm (harvested easily by filtra-
tion), or those forming aggregates/mats, can be a viable
option, as they help to significantly reduce the harvest-
ing cost (Chen et al. 2011; Hori et al. 2002). Another
alternative is the use of a mixture of microalgal consortia
having both unicellular and filamentous strains (Chinnasamy
et al. 2010).
The importance of consortia, as against single organ-
isms, has been illustrated by various workers in terms of
survival, biomass production, as well as nutrient removal
(Bhatnagar et al. 2010; Silva-Benavides and Torzillo
2012). Algal consortia can better grow in wastewaters
because the loss of one alga from the consortia may be
compensated by the other algae (Chinnasamy et al.
2010). Hence, there is urgent need to screen the promis-
ing native microalgae from sewage wastewater that not
only have the potential to sequester excessive nutrients
but also can form consortia having wider biomass appli-
cations after harvesting from the contaminated sites.
Therefore, the present study was carried out to screen
and evaluate the potential of microalgal consortia from
native environment and available germplasm (filamentous
and unicellular) in terms of nutrient removal, improvement of
water quality and biomass production in wastewater under
outdoor conditions.
Materials and methods
Wastewater collection and analyses
Wastewater was collected in clean plastic jars from the
channel at Indian Agricultural Research Institute, New
Delhi. They were transported to laboratory, stored at 4 °C,
and used for further study. Quantification of physicochem-
ical parameters was carried out using the standard methods
J Appl Phycol
for water and wastewater (APHA 2005), and the observa-
tions are tabulated in Table 1.
Isolation of microalgal strains
The native microalgae in wastewater were isolated and puri-
fied after enrichment with modified Bold’s Basal Medium
(BBM) (Starr and Zeikus 1993) supplemented with or without
nitrogen. The composition of the enrichment medium is given
in Supplementary Table 1. For enrichment, medium and
wastewater were taken in 1:1 ratio in 250-mL flasks. Flasks
were incubated at 25±2 °C and 75 μmol photons m−2 s−1 for
3 weeks. Serial dilution technique and streaking method on
BBM agar plates were used for isolation of microalgae from
wastewater, followed by their purification, by picking up
single colonies. The purity of the isolated strains was con-
firmed microscopically. The isolated microalgae were identi-
fied by using standard keys: Cyanobacteria (Komárek and
Anagnostidis 2005), Chlorococcales (Komárek and Fott
1983), and Bacillariophyta (Round et al. 1990).
Development of consortia
Equal amounts of different microalgal strains in terms of
chlorophyll content (0.1–0.2 μgmL−1) were mixed and
allowed to acclimatize under outdoor conditions in modified
Bold’s Basal Medium for consortia development. Three types
of algal consortia, i.e., consortia of native filamentous micro-
algal strains (MC2), native unicellular microalgal strains
(MC3), and selected microalgae from germplasm (MC1), were
used in the present investigation. The details of the microalgal
strains in the various consortia are given in Table 2.
Growth kinetics of consortia and acclimatization
Growth kinetics of consortia was studied in Bold’s Basal
medium in outdoor environment (ex situ) for acclimatiza-
tion. Chlorophyll content was determined spectrophotomet-
rically using methanolic extracts, using the standard
equation of Lichtenthaler (1987). Initial concentration of
all the consortia was kept constant (0.14–0.20 μgmL−1).
Experimental design
Twenty-day-old consortia were centrifuged, repeatedly washed
with distilled water to remove nutrients, and used as inoculum.
Initial chlorophyll concentration was kept constant for all con-
sortia (1.8–2 μg mL−1). Raw sewage water without any inoc-
ulum was kept as control. The experiment was conducted
under outdoor conditions in a net house, using beakers
(1,000 mL) containing 800 mL of sewage wastewater. They
were covered with a transparent film with pores for aeration
and kept at atmospheric night/day temperature and light
J Appl Phycol

intensity (ranged between 17±2–36±3 °C and 420±100– Table 1 Physicochemi-

cal characteristics of Parameter Characteristics
1,760±400 μmol photons m−2 s−1, respectively). The inoculat-
raw sewage water
ed microalgal consortia were harvested at 6 h and 2, 4, 6, 8, 10, pH 8.2
and 14 day time intervals for biomass analysis, and cell-free Salinity 804±2.0
filtrate was taken for the analyses of various physiochemical EC 1,576±5.0
parameters. The experiment was performed twice in triplicates. TDS 1,120±4.0
DO 0.38±0.029
Analytical procedures BOD 2.4±0.001
COD 2,150±50
All the physicochemical parameters, viz, alkalinity, acidity, Alkalinity 223.33±2.89
chloride, carbonate, bicarbonate, calcium, hardness, free Free CO2 13.20±0.29
CO2, dissolved oxygen (DO), biological oxygen demand The units of all the Chloride 273.59±2.96
(BOD), chemical oxygen demand (COD), NO3-N, NO2-N, parameters are in Total hardness 396.67±5.77
NH4-N, and PO4-P, were analyzed using standard methods mg L−1 except pH, bi- Bicarbonate 0.95±0.05
for water and wastewater (APHA 2005). Percent removal was carbonate (meq L−1), Ca Ca and Mg 9.25±0.05
and Mg (meq L−1) and
calculated by the following formula: EC (μS cm−1). Values NO3-N 83.714±0.862
are given as mean of NH4-N 21.067±0.189
ððC0
C Þ=C0 Þ  100 ð1Þ n samples ± SD, PO4-P 3.147±0.015
where n=6
where C0 is the initial concentration of nutrient in wastewater;
C, concentration of nutrient left in wastewater after treatment.
For dry cell weight (DCW) estimation, 10 mL of homog-
day in all the consortia, and thereafter, a plateau or drop
enized culture was filtered through pre-weighed Whatman
was observed, all the three consortia were analyzed period-
no. 41 (ashless) and dried at 60 °C until constant weight was
ically and harvested at the 20th day for analyses.
achieved. DCW was calculated by subtracting the initial
filter paper weight from the final weight of filter paper
containing dried microalgae and was expressed as g L−1. Nutrient removal in wastewater treatment

The NO3-N removal efficiency of different microalgal con-

Statistical analyses
The statistical analyses were performed using the software
Statistical Package for Social Sciences (version 10.0). One-
way analysis of variance (ANOVA) was used to evaluate the
differences among the treatments. The triplicate sets of data
were evaluated in accordance with the experimental design
(completely randomized design) with ANOVA. The com-
parisons between the different means were made using post
hoc least significant differences calculated at P level of 0.05
(5 %) and represented as critical differences values in tables.
SD values are depicted in the graphs as bars. DMRT analysis
was depicted as alphabets (upper/lower case) in tables, with
a/A representing highest values. Similar alphabets indicate
nonsignificantly (p<0.05) different mean values.
Results and discussion
Development and optimization of consortia
All the consortia showed S-shaped growth curve, and mi-
croscopic analyses revealed harmonious growth of the part-
ners in consortia (Supplementary Fig. 1; Fig. 1). Since
maximum chlorophyll content was recorded on the 20th
sortia from sewage water has been illustrated in Fig. 2a. On
the second day, consortium MC2 was able to remove 63.4 %
of NO3-N, while MC1 and MC2 were able to remove only
30.7 and 27.1 % of NO3-N (Fig. 2a). Highest NO3-N removal
of 90 % was observed in MC2-inoculated sewage water on the
tenth day, followed by MC3 and MC1 (82 and 80 %, respec-
tively). In terms of NH4-N, MC2 was able to remove 73 % of
NH4-N from sewage water as compared to 18 and 25 % by
MC1 and MC3 at 6 h (Supplementary Table 2). However,
there was 100 % removal of NH4-N in all the consortia-
inoculated sewage wastewater on the tenth day (Fig. 2b).
The present investigation revealed that MC2 (native fil-
amentous consortia) removed 63 % of NO3-N on the second
day, as compared to two other consortia. This consortium
was also able to remove 100 % of NH4-N and 90 % of NO3-
N from the sewage wastewater on the tenth day. This can be
attributed to the presence of more microalgal strains belong-
ing to the Cyanophyta in MC2 consortia, which are known
to be efficient scavengers of ammoniacal N. The order of
utilization of nitrogen sources by cyanobacteria is, in gen-
eral, NH4+ >NO−3 >N2, and when NH4+ is available, cyano-
bacteria do not utilize other nitrogen sources, until ammonia
is depleted. It is well known that the uptake of nitrate is a
light energy-dependent process involving nitrate reductase,
an inducible enzyme; cyanobacteria prefer to utilize already
 J Appl Phycol

Table 2 Taxonomic description

of the microalgal consortia Consortia Origin Group Genera Morphotype

MC1 Non-native Cyanophyta Calothrix sp. Filamentous

Cyanophyta Lyngbya sp. Filamentous
Chlorophyta Ulothrix sp. Filamentous
Chlorophyta Chlorella sp. Unicellular
MC2 Native Cyanophyta Phormidium sp. Filamentous
Cyanophyta Limnothrix sp. Filamentous
Cyanophyta Anabaena sp. Filamentous
Cyanophyta Westiellopsis sp. Filamentous
Cyanophyta Fischerella sp. Filamentous
Chlorophyta Spirogyra sp. Filamentous
MC3 Native Chlorophyta Chlorella sp. Unicellular
Chlorophyta Scenedesmus sp. Unicellular
Chlorophyta Chlorococcum sp. Unicellular
Cyanophyta Chroococcus sp. Unicellular

reduced nitrogen sources such as ammonium and exhibit
higher biomass productivity (Paerl 1986; Markou and
Georgakakis 2011). Cyanobacteria also store nitrogen in the
form of cyanophycean granules in their cell. This ability can
provide a competitive edge to the MC2 consortium in remov-
ing 90 % of NO3-N from the sewage wastewater, as compared
to the other consortia. Among the microalgae employed for
nutrient removal, Chlorella, Scenedesmus and Spirulina are
most commonly observed algae in wastewaters, but Spirogyra
has also been reported (Aslan and Kapdan 2006).
The comparative PO4-P removal efficiency (percent re-
moval) of different microalgal consortia in sewage water is
illustrated in Fig. 2c. At 6 h, MC1, MC2, and MC3 were
able to remove 83, 67, and 53 % of PO4-P, respectively
Fig. 1 Microphotographs
illustrating the microalgal a Ly
diversity in the consortia on the
20th day. a MC1 comprising Ly
Lyngbya sp., Chl Chlorella sp.,
Ca Calothrix sp., U Ulothrix sp.
b MC3 comprising Chl
Ca
Chlorella sp., Chr Chroococcus
sp., Sc Scenedesmus sp. c, d
MC2 comprising L Limnothrix
sp., Ph Phormidium sp., An 20 µm
(Supplementary Table 2). On the secondday, MC2 was able
to remove 90 % of PO4-P, while MC1 and MC2 were able to
remove only 88 and 87 % of PO4-P, respectively. On the
fourth day onwards, no significant differences in the PO4-P
removal potential of the consortia was recorded (Fig. 2c).
The investigation showed that MC2 was more efficient
than MC1 and MC3 in sequestration of phosphorus from the
sewage wastewater, which can be attributed to the presence
of more cyanobacteria, which can store excess of P in the
form of polyphosphate bodies (Markou and Georgakakis
2011). Hence, mixtures/consortia of filamentous and unicel-
lular strains are commonly used to combine the advantages
of high biomass and proliferation of unicellular forms with
gas vacuole containing filamentous, providing buoyancy
b
U Chl
Chl
Chr Sc
Ch
20 µm

Anabaena sp., S Spirogyra sp.,

F Fischerella sp., W
Westiellopsis sp c d
Ph W
An
L
S An
W

20 µm 20 µm
J Appl Phycol

a In our investigation, pH of raw sewage was 8.2 which

100 increased to pH 8.5±0.2 at 6 h and, then on the second day,
Percent Removal of NO3-N

80
to pH10.2. The observed increase in pH is due to the photo-
synthetic activity of microalgae which plays an important role
60 in wastewater treatment, as a result of cycling of different
forms of N and P present in wastewater (Silva-Benavides
40
and Torzillo 2012) At higher pH (≥9), free ammonia begins
20 to dominate over ammonium ion, and PO4− is known to
precipitate as insoluble salts (Markou and Georgakakis
0 2011; Silva-Benavides and Torzillo 2012). However, in the
0d 2d 4d 6d 8d 10d
present investigation, 83.3, 67.4, and 53.9 % of available P
Time (d)
were removed by MC1, MC2, and MC3, respectively, from
b the sewage wastewater at 6 h when pH was more or less the
120 same as that of initial (0 day). The uptake of available P is an
Percent Removal of NH4-N

100 active process which operates gradually over time. In our

study, by the second day, up to 80 % removal was achieved,
80
after which a plateau was reached. Hence, the observed higher
60 pH (10.2) on the second day did not have any significant effect
40 on removal of P by algae. Similarly, 80–85 % NH4-N was
20
removed by different consortia by the second day, and any
volatilization of ammonia due to high pH may be the reason
0
0d 2d 4d 6d 8d 10d
for any further N losses.
Time (d)

c
120 a
Percent Removal of PO4-P

120
100
Percent reduction in BOD

100
80
80
60
60
40
40
20
20
0
0d 2d 4d 6d 8d 10d 0
Time (d) 0d 2d 4d 6d 8d 10d
Time (d)
Fig. 2 Percent removal of a NO3-N, b NH4-N, and c PO4-P from sewage
wastewater by various microalgal consortia at different time intervals.
Raw sewage wastewater (──○──), MC1 (—▲—), MC2 (– –□– –), and
b
80
MC3 (—◊—) are shown. Error bars indicate standard deviation
Percent reduction in COD

60

mechanism and a matrix or the unicellular forms. Such 40

consortia have shown to perform better in phytoremediation
experiments. Chinnasamy et al. (2010) reported that micro- 20
algal consortia comprising both native unicellular and fila-
mentous strains were able to remove 99.8 % of NO3-N and 0
96.6 % of PO4-P within 72 h when grown in sterilized carpet 0d 2d 4d 6d 8d 10d
industry effluent under elevated CO2 levels (6 %) and at 25 °C. Time (d)
In an another study, 80 % nitrogen removal was recorded
Fig. 3 Percent reduction in a BOD and b COD of sewage wastewater by
from municipal wastewater in which co-culturing of Plank- various microalgal consortia at different time intervals. Raw sewage waste-
tothrix isothrix and Chlorella vulgaris was undertaken (Silva- water (──○──), MC1 (—▲—), MC2 (– –□– –), and MC3 (—◊—) are
Benavides and Torzillo 2012). shown. Error bars indicate standard deviation
 J Appl Phycol

Values given are mean of n samples ± SD, where n=6. Letters in lower case (a, b …) denote rankings in respective column, and letters in upper case (A, B …) denote rankings in respective rows
CD (5 %)
BOD and COD removal

0.202
0.169
0.267
0.269
–
A gradual decrease in BOD was observed in all the consor-
tia studied (Fig. 3a). On the second day, highest removal of

4.467±0.115 b, B
4.467±0.115 b, B
3.500±0.100 c, B
5.340±0.020 a, B
81 % of BOD was observed in MC3, followed by MC2
(77 %). While on the sixth day, highest reduction in BOD
(88 %) was found to be in MC2-inoculated sewage water. At

14th day
the end of experiment (14 days), all the consortia showed

0.164
the same efficiency (99 %) for BOD removal (Supplemen-
tary Table 2). COD removal also showed a similar trend as
that of BOD. On the sixth day of study, there was a 65 %

4.173±0.300 b, A

4.413±0.012 b, B
4.067±0.115 b, C
5.193±0.012 a, B
reduction in COD in MC1 and MC3 and 61 % reduction in
MC2-inoculated sewage water (Fig. 3b). Highest reduction

10th day
in COD up to 88 % was observed in MC1, while MC2 and

0.274
MC3 were able to reduce 87 % of COD in sewage water on
the 14th day of study (Supplementary Table 2). Riano et al.
(2011) demonstrated that consortia of microalgae isolated

8.000±0.400 b, A
8.000±0.400 b, A
9.000±0.200 a, A
3.633±0.153 c, B
from aerobically treated swine slurry can remove 70 % of
COD from wastewater from fish processing unit after 5 days.

Table 3 Influence of different microalgal consortia on dissolved oxygen (mg L−1) of wastewater at different time intervals
In another report, maximum reduction (89.29 %) and COD

6th day
(73.68 %) were recorded using Anabaena variabilis and Ana-

0.335
baena oryzae, respectively from mixed industrial wastewater
after 7 days of incubation. El-Bestawy (2008), who also found
that the use of single microalga Tolypothrix ceylonica led to a

3.920±0.106 ab, C
3.800±0.200 b, C
0.399±0.001 c, C

4.067±0.115 a, C
decrease in total dissolved solids (TDS) to 13.76 % in mixed
domestic industrial wastewater in 7 days.
4th day

Water quality improvement 0.177

All the three consortia were able to improve water quality,

1.327±0.110 b, D
1.617±0.029 a, D
1.527±0.110 a, D
0.413±0.023 c, C

but to different extents. There was a significant (p<0.05)

increase in DO levels in all the treatments (Table 3). Max-
imum increase in DO levels was observed on the sixth day
of study in MC1 as 9 from 0.38 mg L−1, followed by MC2
2nd day

0.138

and MC3 (Table 3). On the sixth day of study, MC1, MC2,
and MC3 were able to reduce total hardness to 223, 258, and
243 mg L−1, respectively, from 396.67 mg L−1 (Table 4).
0.393±0.012 b, C
0.460±0.104 b, E
0.420±0.035 b, E
0.827±0.046 a, E

Maximum reduction in hardness was observed in MC1, i.e.,

149.33 mg L−1 on the 14th day (Table 4). In all the treat-
ments, free CO2 was completely removed on the secondday
0.103

(data not shown). Alkalinity was reduced to 81, 131.67, and

6h

82.50 mg L−1 from 223.33 mg L−1 by MC1, MC2, and

MC3, respectively, on the second day. Highest reduction in
alkalinity to 44 mg L−1 was observed in MC3-inoculated
0.38±0.01 C
0.38±0.01 E
0.38±0.01 E
0.38±0.01 F

sewage water followed by MC1 and MC2 to 45 and

58.33 mg L−1, respectively, on the 14th day of study
0 day

(Table 5). In the present study, a similar trend was ob-

served in TDS, electrical conductivity (EC), and salinity
Type of consortia

(Fig. 4a–c). Maximum decrease in TDS level was

achieved on second day in MC1 (29.1 %) followed by
CD (5 %)
SW MC1
SW MC2
SW MC3

MC2 and MC3 (Fig. 4a). However, MC2 was able to

Control

decrease TDS (28 %) to 806 mg L−1 on the sixth day

(Fig. 4a). Highest reduction in EC to 1,119 μScm−1 and
Table 4 Influence of different microalgal consortia on total hardness (mg L−1) of wastewater at different time intervals

Type of consortia 0 day 6h 2nd day 4th day 6th day 10th day 14th day CD (5 %)
J Appl Phycol

Control 396.67±5.77 A 398.67±0.58 a, A 399.67±0.58 a, A 338.00±1.00 a, B 303.67±1.53 a, C 237.67±1.53 a, D 217.67±1.53 a, E 2.53

SW MC1 396.67±5.77 A 372.33±0.58 b, B 317.00±2.00 c, C 233.00±1.00 b, D 223.00±1.00 d, E 175.00±1.00 c, F 149.33±2.31 d, G 2.56
SW MC2 396.67±5.77 A 359.67±0.58 d, B 317.00±1.00 c, C 209.67±0.58 c, D 258.00±1.00 b, E 167.33±1.15 d, F 165.67±2.08 c, F 2.44
SW MC3 396.67±5.77 A 364.33±3.79 c, B 326.33±2.08 b, C 231.67±2.08 b, D 243.00±1.00 c, E 190.33±0.58 b, F 170.67±1.15 b, G 2.95
CD (5 %) – 2.60 1.90 1.80 1.83 0.76 2.58 –

Values given are mean of n samples ± SD, where n=6. Letters in lower case (a, b …) denote rankings in respective column, and letters in upper case (A, B …) denote rankings in respective rows

Table 5 Influence of different microalgal consortia on alkalinity (mg L−1) of wastewater at different time intervals

Type of consortia 0 day 6h 2nd day 4th day 6th day 10th day 14th day CD (5 %)

Control 223.33±2.89 B 223.30±3.99 ab, B 231.67±2.89 a, A 228.33±2.89 a, A 228.33±2.89 a, A 163.83±0.29 a, C 158.33±2.89 a, D 4.57
SW MC1 223.33±2.89 A 223.33±7.64 ab, A 81.00±1.00 d, B 64.83±0.29 c, C 65.17±0.29 c, C 62.17±2.02 c, C 45.00±5.00 b, D 5.47
SW MC2 223.33±2.89 A 227.67±2.52 a, A 132.67±6.43 b, B 72.67±2.52 b, C 71.83±2.275 b, C 58.67±0.29 d, D 58.33±2.89 a, D 5.43
SW MC3 223.33±2.89 A 221.33±5.51 b, A 89.50±0.50 c, B 65.17±0.29 c, C 73.50±1.50 b, C 70.17±0.29 b, D 44.00±1.00 b, E 3.35
CD (5 %) – 4.05 6.18 3.46 3.38 1.71 2.97 –

Values given are mean of n samples ± SD, where n=6. Letters in lower case (a, b …) denote rankings in respective column, and letters in upper case (A, B …) denote rankings in respective rows
 J Appl Phycol

a 1.2
1200
1
1100

DCW (g L-1)
0.8
TDS (mg L-1)

1000 0.6

900 0.4

0.2
800
0
700 0d 2d 4d 6d 8d 10d
0d 2d 4d 6d 8d 10d
Time (d) Time (d)

b Fig. 5 Biomass production (DCW) of the microalgal consortia grown

1700
on sewage wastewater at different time intervals. Raw sewage waste-
water (──○──), MC1 (—▲—), MC2 (– –□– –), and MC3 (—◊—)
1600 are presented. Error bars indicate standard deviation
1500
EC (µS cm-1)

1400 Biomass production

1300
All the three consortia were able to proliferate in sewage
1200
water. Highest biomass (1.07 gL−1) was obtained from MC2
1100 on the sixth day. However, MC1 and MC3 were able to
1000 produce biomass of 0.90 and 0.94 g L−1, respectively
0d 2d 4d 6d 8d 10d (Fig. 5). After sixth day, a decrease in biomass was observed
Time (d)
in all the consortia. A decrease in biomass after 6 days in all
c the consortia revealed the end of exponential phase. The
900 observed differences in biomass generated using different
microalgal consortia might be because of their differential
800 response under outdoor environmental conditions, viz. light,
Salinity (mg L-1)

temperature etc. However, an increase in biomass was ob-

700
600
500
0d 2d 4d 6d 8d 10d
Time (d)
Fig. 4 Comparative evaluation of a TDS, b EC, and c salinity levels in
various microalgal consortia-inoculated sewage wastewater at different
time intervals. Raw sewage wastewater (──○──), MC1 (—▲—),
MC2 (– –□– –), and MC3 (—◊—) are presented. Error bars indicate
standard deviation
salinity to 569 mg L−1 on second day were observed using
MC1. However, a significant (p<0.05) increase in EC and
salinity was observed in MC1- and MC3-inoculated sewage
wastewater on fourth day, which, however, decreased there-
after. The maximum reduction in EC and salinity (Fig. 4b–c)
was observed on the sixth day in MC2-inoculated sewage
water (1,130 μScm−1 and 582 mg L−1, respectively). In the
present investigation, the highest increase in DO and highest
reduction in TDS, EC and salinity were observed on sixth
day, when MC2 consortium of native filamentous strains was
employed.
served with increase in incubation time in sewage wastewater,
which indicated that the sewage water was a conducive
growth medium for all the microalgal consortia employed in
the present investigation.
The present study showed that native filamentous con-
sortium was able to produce the highest biomass of 1.07 g
L−1 on sixth day. There are a few reports which demonstrate
that native microalgal strains adapt better to the local envi-
ronment and exhibit faster growth rate compared with those
introduced (Zhou et al. 2012b). Native microalgae are
known to possess better potential to grow in wastewaters
having high carbon, nitrogen, and phosphorus content and
tend to aggregate and exhibit self-sedimentation without
addition of any flocculants (Zhou et al. 2012a). Chinnasamy
et al. (2010) studied that consortia of native microalgal
strains can produce biomass of 1.47 gL−1 on the ninth day
when grown in sterilized carpet mill wastewater with in-
creased CO2 level (6 %) and at 25 °C. Riano et al. (2011)
found that microalgae consortium isolated from swine slurry
can produce 0.555 g L−1 of biomass in 5 days, when culti-
vated in fish processing wastewater in a photobioreactor. In
our study, the consortia developed using microalgae belong-
ing to available germplasm (MC1) was able to maintain its
biomass up to 14 days, after attaining maxima on sixth day,
J Appl Phycol
indicative of the robustness and inherent tolerance potential
to adverse environments of its constituent strains.
From the present investigation, it can be concluded that the
consortium of native filamentous strains (MC2) producing
highest biomass (1.07 mg L−1) and exhibiting highest NO3-
N removal (90 %), complete NH4-N removal and highest
PO4-P (97 %) removal can be considered most promis-
ing, when grown in sewage wastewater under outdoor
conditions. Besides nutrient removal, MC2 was able to
reduce the TDS to the highest level, decrease the level
of EC and salinity and increase the level of DO. Hence,
this consortium of filamentous native strains can be a
promising option in terms of biomass production, nutri-
ent removal and improvement of water quality of sew-
age water; it can be exploited both for in wastewater
treatment, and the generated biomass can be utilized in
various biotechnological applications such as biofuel,
biofertilizer and production of bioactive compounds.
Acknowledgments The first author is thankful to University Grants
Commission, New Delhi, for her fellowship. All the authors are thankful
to the Department of Botany, Panjab University, Chandigarh; Division of
Microbiology, Indian Agricultural Research Institute, New Delhi; and
University of Delhi, Delhi, for providing the budget and research facilities
to carry out the present investigation.
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