Materials Today: Proceedings xxx (xxxx) xxx

Contents lists available at ScienceDirect

Materials Today: Proceedings

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

A comparative study of the growth of microalgae-bacteria symbiotic

consortium with the axenic culture of microalgae in dairy wastewater
through extraction and quantification of chlorophyll
Namita Talapatra, Rahul Gautam, Vaishali Mittal, U.K. Ghosh ⇑
Department of Polymer and Process Engineering, IIT Roorkee Saharanpur Campus, Saharanpur 247001, India

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

Article history: Microalgae-bacteria symbiotic association can provide many competitive advantages over the pure axe-
Available online xxxx nic culture of microalgae to obtain biofuel. This system significantly reduces an important technical con-
straint, including associated capital cost with the infrastructure required for maintaining pure microalgae
Keywords: culture. The current study aims to compare the symbiotic growth of Tetraselmis indica - Pseudomonas
Axenic pure microalgae culture (APMC) aeruginosa consortium with axenic growth of Tetraselmis indica in dairy wastewater (DWW). The extrac-
Microalgae-bacteria consortium tion and optimization of chlorophyll-a were also investigated. The overall maximum chlorophyll-a recov-
Chlorophyll extraction
ery of 29.106 ± 0.045 mg L
1 was obtained through the sonification as extractant method with methanol
Algal biomass
Dairy wastewater
as an extraction solvent. The chlorophyll-a content of consortium cultivated in DWW higher than the
axenic pure microalgae culture (APMC). The maximum biomass yield of the consortium was achieved
as 1454.88 mg L
1, which is much higher than APMC growth in DWW by 38.80%. On the 10th day, the
consortium removed 87.49%, 83.76%, and 79.83% of chemical oxygen demand (COD), total dissolved
nitrogen (TDN), and total dissolved phosphorus respectively. The higher biomass growth and wastewater
nutrient removal exhibited by the symbiotic microalgae-bacteria consortium can be utilized in biofuel
technology.
Ó 2021 Elsevier Ltd. All rights reserved.
Selection and peer-review under responsibility of the scientific committee of the International Confer-
ence on Nanoelectronics, Nanophotonics, Nanomaterials, Nanobioscience & Nanotechnology.

1. Introduction gae cultivation. A consistent rise in the demand for milk and milk
based food items in a large number of countries has led to signifi-
Microalgae can be utilized to obtain a wide variety of metabo- cant progress in the world’s dairy industries. DWW can be used as
lites such as carbohydrates, proteins, food and feed additives, a nutrient supplement for the cultivation of consortium. The asso-
lipids, carotenoids or health vitamins, cosmetics, and energy pro- ciated bacteria prevent microalgae from unwanted bacterial con-
duction [1]. Microalgae can be harnessed to produce various bioac- tamination [2], enhance microalgae growth by releasing certain
tive compounds for commercial use. One of the major obstacles for plant growth-promoting factors and extracellular polymeric sub-
the cultivation of axenic pure microalgae culture (APMC) is the stances [3], and simultaneously treat wastewater by reducing the
high infrastructural cost for maintaining the pure microalgae cul- biological load [4] (Fig. 1).
ture. The microalgae-bacteria consortia approach is useful in bring- Conventional wastewater treatment methods significantly
ing down the cost of cultivation of microalgae in nutrient-rich reduce high operational and capital costs by employing
media. Only few studies have been performed on the cultivation microalgae-bacteria consortia. Theoretically, a microalgae growing
of symbiotic association of microalgae bacteria consortium in dairy industry can be developed with simultaneous treatment of the
wastewater (DWW), which has tremendous potential for microal- dairy effluent. Microalgae pose a simple structure, can grow
rapidly with solar conversion efficiency higher than other terres-
trial plants [5]. The dairy industries are estimated to produce 2.5
⇑ Corresponding author at: IIT Roorkee, Polymer and Process Engineering, IIT to 3 L of wastewater per litre of processed milk, which has to be
Roorkee Saharanpur Campus, Saharanpur, Uttar Pradesh, 247001, India. treated before releasing into the environment [6]. As the nutrient
E-mail address: [email protected] (U.K. Ghosh).

https://doi.org/10.1016/j.matpr.2021.06.227
2214-7853/Ó 2021 Elsevier Ltd. All rights reserved.
Selection and peer-review under responsibility of the scientific committee of the International Conference on Nanoelectronics, Nanophotonics, Nanomaterials,
Nanobioscience & Nanotechnology.

Please cite this article as: N. Talapatra, R. Gautam, V. Mittal et al., A comparative study of the growth of microalgae-bacteria symbiotic consortium with the
axenic culture of microalgae in dairy wastewater through extraction and quantification of chlorophyll, Materials Today: Proceedings, https://doi.org/
10.1016/j.matpr.2021.06.227
N. Talapatra, R. Gautam, V. Mittal et al. Materials Today: Proceedings xxx (xxxx) xxx

Fig. 1. Mechanism of algae-bacteria interactions in wastewater.

load of DWW is significantly high, it can facilitate the growth of the P. aeruginosa was purchased from National Chemical Laboratory
consortium by serving as a nutrient medium. Bacteria enhance the (NCL) Pune, India. The bacterium was grown in an LB nutrient
growth of microalgae which further increases microalgae biomass broth at 30 ± 2° C in a rotary shaker at 120 rpm for 18 h.
productivity. Also, the consortium can reduce the harvesting cost The DWW samples collected from Kwality Ltd., dairy processing
of microalgae biomass significantly. Simultaneously, industrial plant in Saharanpur, India and stored at 4° C immediately to elim-
dairy wastewater can be treated as the targeted industry releases inate decomposition of the substrate prior to experiment. Table 1
effluent with high organic matter content and high COD [7]. shows the physicochemical properties of DWW. The initial pH of
One of the valuable bioactive compounds derived from microal- the DWW was 4.0, which was later maintained to 7.5.
gae biomass is chlorophyll. Microalgae growth can be indicated
through the concentration of chlorophyll-a. The chlorophyll-a con- 2.2. Experimental set-up for culture
centration also indicates the growth phase of the photosynthetic
process in microalgae. The measurement of only dry cell weight Experiments were carried out with a working volume of 300 ml
to study microalgae growth in wastewater is inappropriate as the in four 500-mL Erlenmeyer flasks. Microalgae-bacteria inoculum
wastewater also contains organic and inorganic content that con- was cultivated in BG11 medium as control and DWW while APMC
tributes to the dry weight. The best studied mechanism to measure was maintained in BG11 medium as control and DWW. All the
microalgae growth in wastewater is the determination of three cultures were cultivated under a light intensity of
chlorophyll-a content. 130 lmol/(m2 s) with a 16 h/8h light/dark cycle at 28 °C for
The present study was undertaken for extraction and optimiza- 10 days. A system of continuous air sparging into the system was
tion of chlorophyll-a content from microalgae. The extraction pro- maintained through aquarium air pump (Sebo SB-548A) (Fig. 2).
cess involves various unit operations such as grinding, ultrasound
or sonification, homogenization. Also, the various organic solvent 2.3. Analytical methods
used to extract chlorophyll-a, such as acetone, methanol, ethanol,
Chloroform, etc. Su et al. [8] and Halim et al. [9] found that metha- 2.3.1. Dry weight
nol and acetone to be efficient extraction solvents. So chlorophyll A filter paper of pore size 0.45 lm was dried for 6 h in a dryer at
extraction efficiency was compared using acetone and methanol 105 ± 5 °C to assess dry cell weight and then weighted by an elec-
as an extracting organic solvent. A comparison was made between tronic microbalance (W1). After filtering the 10 ml sample, the fil-
efficiency achieved through solvent suspension method for 24 h in ter paper dried to constant weight (W2). The dry weight of samples
dark and cell disruption achieved through sonification. In addition, was estimated by subtracting the initial weight from the final
the growth of Tetraselmis indica in the presence of consortium in weight. The dry cell weight (mg L
1) and biomass productivity
DWW were monitored. (mg L
1day
1) were calculated from Eqs. (1) and (2), where T
was culture time in days (Figure 3).
mg 
Dry cell weight ¼ ðW2
W1Þ=V ð1Þ
L
2. Materials and methods mg
Biomass productiv ity l ¼ Dry cell weight=T ð2Þ
d
2.1. Microalgae, bacteria culture, and the sample collection

T. indica was procured from National Facility for Marine

Cyanobacteria Centre (NFMC), Tamil Nadu. Algae inoculums were
Table 1
cultivated for 7 days under 130 lmol/(m2 s) light intensity with
Physiochemical characteristics of DWW.
a 16 h/8h light/dark cycle in 500 ml BG-11 medium present in
1000 ml conical flasks. The composition of BG11 medium per litre Characteristics Value
was comprising of NaNO3 0.75 g, K2HPO4 0.04 g, CaCl22H2O COD 1953.8 ± 161.5
0.036 g, MgSO4 0.04 g, NaCO3 0.020 g, Mg-EDTA 0.001 g, Citric acid TDN 78.8 ± 6.3
TDP 15.3 ± 2.1
0.0056 g, Ferric Ammonium Citrate 0.006 g, and trace metals 1.0 ml
2
N. Talapatra, R. Gautam, V. Mittal et al. Materials Today: Proceedings xxx (xxxx) xxx

Fig. 2. Experimental set-up of the APMC and consortium system.

Fig. 3. Changes in dry weight of Tetraselmis indica under APMC and Consortium system.

2.3.2. Measurement of water quality 2.3.3.1. Solvent suspension method (SSM). 10 ml sample of consor-
The dairy wastewater was filtered with 0.45 lm membrane fil- tium in DWW was centrifuged (Remi R-24, Mumbai, India) for
ters. The filtrate was used for the water quality parameters deter- 10 min at 5000 rpm. The supernatant was discarded, and the pellet
mination. COD, TDN, and TDP were determined by the Hanna was washed twice with distilled water and centrifuged. The pellet
multiparameter photometer (HI839800). The chromotropic acid left was resuspended with 10 ml of 90% acetone and then stored
and vanadomolybdophosphoric acid methods were used for the for 24 h in darkness at 4 °C. After 24 h, the centrifugation of the col-
TDN and TDP determination, respectively. And dichromate method lected suspension was performed at 5000 rpm for 10 min. The
was used to measure COD. absorbance of the supernatant at wavelengths of 645, 652, 663,
and 665 nm was recorded in a UV spectrophotometer using 90%
of acetone as a blank [10].
2.3.3. Concentration of chlorophyll-a Similarly, centrifugation of a 10 ml sample of consortium in
The UV–visible spectrophotometer (Shimadzu UV-1800, Kyoto, DWW was carried out at 5000 rpm for 10 min, and the supernatant
Japan) was utilized to measure the concentration of chlorophyll-a was removed. The pellet left in the bottom of the centrifuge tube
by measuring the optical densities of each sample. Four chlorophyll was washed two times with distilled water and centrifuged. The
extraction procedures were employed to get a superior result. Also, pellet was resuspended in 90% methanol (10 ml) and then stored
methanol and acetone were used as extracting solvent. for 24 h in darkness at 4 °C. after 24 h. The suspension was cen-

3
N. Talapatra, R. Gautam, V. Mittal et al. Materials Today: Proceedings xxx (xxxx) xxx

trifuged at 4000 rpm for 15 min. The absorbance of the supernatant 2.4. Data analysis
was measured at wavelengths of 645, 652, 663, and 665 nm in an
ultraviolet spectrophotometer using 90% of methanol as a blank All the experiments have been performed in triplicates. And the
[11]. experimental results have been shown as the means with standard
deviations. Origin 8.5.0 software was used to analyse the collected
2.3.3.2. Sonification method (SM). Centrifugation of a 10 ml sample data.
of consortium in DWW was performed for 10 min at 5000 rpm.
After discarding the supernatant, pellet was washed twice with
3. Results and discussions
distilled water and centrifuged. The pellet obtained was resus-
pended in 10 ml of 90% acetone. The extraction process was per-
3.1. Biomass production
formed using probe ultrasonic for 2 min with 25 mm tipped
probe (Miko India GOC35, India) with continuous output (approx-
The changes in dry cell weight were determined for Tetraselmis
imately 19 W/cm2), the output controller is set to seven. Sample
indica under APMC and consortium system (Fig. 4). The dry cell
bottles were covered with aluminium to avoid chlorophyll degra-
weight patterns of the APMC and consortium system in DWW
dation due to light and put in a probe ultrasonic in a predefined
were compared with BG11 media as a control. The presence of
state [12]. The sonication vessel is kept in a beaker containing ice
Pseudomonas aeruginosa in the symbiotic environment has also
to maintain the temperature below 35 °C. The absorbance of the
enhanced the weight of the dry cells. The biomass growth of sym-
supernatant at wavelengths of 645, 652, 663, and 665 nm was
biotic association of the consortium system in DWW was 1454.8
recorded in a UV spectrophotometer using 90% of acetone as a
8 ± 40.34 mg L
1 showed increased maximum biomass by
blank [12].
38.80% compared with the APMC system. Bacteria support microal-
10 ml of the sample of consortium in DWW was centrifuged for
gae growth by providing CO2, phytohormones, remineralized
10 min at 5000 rpm. After discarding the supernatant, pellet was
macro, and micronutrients; in return, microalgae support bacteria
washed twice with distilled water and centrifuged. The pellet
growth by providing O2 and organic compounds to bacteria.
obtained was resuspended in 10 ml of 90% methanol. The extrac-
The value of the biomass growth of Tetraselmis indica in con-
tion process was performed using probe ultrasonic for 2 min with
sortium was higher than the biomass concentration of Chlorella
25 mm tipped probe (Miko India GOC35, India) with continuous
Vulgaris, 792 mg L
1, which Shi et al. in DWW [14]. The growth
output (approximately 19 W/cm2), the output controller is set to
of the consortium was found to be 1420.83 ± 38.76 mg L
1 on
seven. Sample bottles were covered with aluminium to avoid
the 10th day of cultivation in DWW, indicating the effective uti-
chlorophyll degradation due to light and put in a probe ultrasonic
lization of organic compounds by Tetraselmis indica. While the
in a predefined state [12]. The sonication vessel is kept in a beaker
experiment was continued for further two days, no such improve-
containing ice to maintain the temperature below 35 °C. The absor-
ment was detected in the dry cell weight as stationary phase was
bance of the supernatant at wavelengths of 645, 652, 663, and
attained. Maximum biomass concentration was achieved for all
665 nm was recorded in a UV spectrophotometer using 90% of
the rest cases on the 8 days of the cultivation period. However,
methanol as a blank [12].
presence of excess nutrients above the optimum concentration
The concentration of Chlorophyll-a (mg L
1) was determined by
reduce the growth of the microalgae [15].
the following Eqs. (3) for acetone as an organic solvent:
mg 
Chla ¼ 12:7  ðODÞ663
2:69  ðODÞ645 ð3Þ 3.2. Changes in the concentration of COD, TDN, and TDP
L

1
The concentration of Chlorophyll-a (mg L ) was determined by The changes of in concentrations of TDN, TDP and COD were
the following Eqs. (4) for methanol as an organic solvent [13]: measured for Tetraselmis indica - Pseudomonas aeruginosa consor-
mg 
tium system and Tetraselmis indica APMC system (Fig. 5.). The
Chla ¼ 16:75  ðODÞ665
9:16  ðODÞ652 ð4Þ
L COD concentration on the 10th day of the consortium system in
DWW was lesser than that of APMC in DWW. Similar observations

Fig. 4. (a) Initial Concentration (b) concentration change in APMC system and (c) concentration change in consortium system after 10 Days in (i) COD (ii) TDN and (iii) TDP.

4
N. Talapatra, R. Gautam, V. Mittal et al. Materials Today: Proceedings xxx (xxxx) xxx

Fig. 5. Absorbance in UV Spectrophotometer for different chlorophyll extraction methods.

were observed for TDN and TDP concentrations. The concentration

of COD decreased from 1953.8 ± 161.5 mg L
1 to 244.4 ± 7.1 mg L
1
in the consortium system representing 87.49% of removal effi-
ciency. The APMC system exhibited lesser COD removal rate from
1953.8 ± 161.5 mg L
1 to 443.5 ± 11.2 mg L
1, which is only
77.30% of removal efficiency. (Fig. 4.i).
Similar results were observed for TDN and TDP concentrations
of consortium system as compared to the APMC system. The TDN
value of consortium were measured with removal rate of 83.76%
from 78.8 ± 6.3 mg L
1 to 12.8 ± 0.8 mg L
1 while APMC system
observed 75.12% removal rate (Fig. 4.ii). Also, TDP value of consor-
tium were measured with removal rate of 79.83% from 15.3 ± 2.1
mg L
1 to 3.1 ± 0.2 mg L
1 while APMC system observed 71.53%
removal rate (Fig. 4.iii). These findings showed the strong synergy
Fig. 6. Chlorophyll-a (mg L
1) concentration through sonification method in (a)
in removing COD, TDN, and TDP under Tetraselmis indica - Pseu-
Tetraselmis indica in control, (b) Tetraselmis indica - Pseudomonas aeruginosa in
domonas aeruginosa consortium system. Under the consortium sys- control, (c) Tetraselmis indica APMC in DWW, and (d) Tetraselmis indica -
tem, the presence of Pseudomonas aeruginosa contributed to a Pseudomonas aeruginosa consortium in DWW.
higher removal rate of COD, TDN, and TDP in DWW compared to
the APMC system [16].
Table 2
Chlorophyll-a content of different microalgae species involving different solvents.
3.3. Relative quantification of Chlorophyll-a
Microalgae Solvent Chlorophyll-a content References
(mg L
1)
The growth of a consortium of mixed culture of Tetraselmis
Chlorella sp. Methanol: 17.19 [17]
indica - Pseudomonas aeruginosa has been studied in comparison
Hexane
with the Tetraselmis indica APMC cultivated in DWW and standard Microalgae Acetone 3.58 [18]
BG11 media. Phototrophic growth of microalgae can be indicated Chlorella Methanol 22.0 [13]
through chlorophyll-a content. The maximum concentration of pyrenoidosa
chlorophyll-a of 29.106 ± 0.045 mg L
1 was calculated in a batch Chlorella sp. BWY-1 Acetone 9.37 [16]
Chlorella sp. Acetone 16.3 [19]
culture of consortia of Tetraselmis indica-Pseudomonas aeruginosa
Tetraselmis indica Methanol 29.106 Current
in DWW and 20.42 ± 0.29 mg L
1 for a pure culture of Tetraselmis study
indica maintained in DWW. The chlorophyll-a content of the con- Two microalgal Methanol 19.17–25.17 [20]
sortium is 42.54% higher than APMC in DWW (Fig. 6.). A similar consortia
trend of dry cell weight to that chlorophyll-a content could be
observed. The dry cell weight was higher for the consortium sys-
tem as compared to APMC. The absorbance in the sonification method was highest with
To measure the chlorophyll-a content of methanol and ethanol methanol as an organic solvent compared to the solvent suspen-
as an extraction solvent, absorbances were recorded at wave- sion method involving acetone and methanol as an organic extract-
lengths of 645, 652, 663, and 665 nm in a UV spectrophotometer. ing solvent.
5
N. Talapatra, R. Gautam, V. Mittal et al. Materials Today: Proceedings xxx (xxxx) xxx

Mechanical disruption is required to maximize the recovery of towards sustainable biofuel production, Bioresour. Technol. 221 (2016) 455–
460, https://doi.org/10.1016/j.biortech.2016.09.070.
pigments from microalgae cells. In comparison to other reported
[5] J.K. Nayak, U.K. Ghosh, Post treatment of microalgae treated pharmaceutical
chlorophyll-a content, a higher concentration of chlorophyll-a wastewater in photosynthetic microbial fuel cell (PMFC) and biodiesel
obtained in methanol is 29.06 mg L
1 as recorded in Table 2. Probe production, Biomass Bioenergy 131 (2019), https://doi.org/10.1016/j.
sonication facilitates the optimum extraction of chlorophyll-a from biombioe.2019.105415 105415.
[6] N.P. Shete Bharati, Shinkar, Dairy industry wastewater sources, characteristics
microalgae. Methanol is more effective than acetone. The pigment & its effects on environment, Int. J. Curr. Eng. Technol. 3 (5) (2013) 1611–1615.
analysis is conducted just after the extraction while precautions [7] E. Bazrafshan, F. Kord Mostafapour, M. Alizadeh, M. Farzadkia, Dairy
need to maintain to avoid sample heating during sonication. wastewater treatment by chemical coagulation and adsorption on modified
dried activated sludge: a pilot-plant study, Desalin. Water Treat. 57 (18) (2016)
8183–8193, https://doi.org/10.1080/19443994.2015.1018331.
[8] S. Su, Y. Zhou, J.G. Qin, W. Yao, Z. Ma, Optimization of the method for
4. Conclusion
chlorophyll extraction in aquatic plants, J. Freshw. Ecol. 25 (4) (2010) 531–538,
https://doi.org/10.1080/02705060.2010.9664402.
This study demonstrated that the presence Pseudomonas aerug- [9] R. Halim, A. Hosikian, S. Lim, M.K. Danquah, Chlorophyll extraction from
inosa enhances the growth of Tetraselmis indica, which resulted in microalgae: A review on the process engineering aspects, Int. J. Chem. Eng.
(2016), https://doi.org/10.1155/2010/391632.
increase of chlorophyll-a concentration as compared to APMC. [10] C.J. Lorenzen, Determination of Chlorophyll and Pheo-Pigments:
The biomass content of Tetraselmis indica- Pseudomonas aeruginosa Spectrophotometric Equations, Limnol. Oceanogr. 12 (2) (1967) 343–346,
consortium is highest in DWW. The dry cell weight of Tetraselmis https://doi.org/10.4319/lo.1967.12.2.0343.
[11] H.K. Lichtenthaler, C. Buschmann, Chlorophylls and Carotenoids:
indica in control was 825.72 ± 32.86 mg L
1. This indicates that Measurement and Characterization by UV-VIS Spectroscopy, Curr. Protoc.
DWW is a good source of nutrient media to be used for microalgae Food Anal. Chem. 1 (1) (2001), https://doi.org/10.1002/
cultivation. Compared to the axenic culture of Tetraselmis indica, 0471142913.faf0403s01.
[12] D. Simon, S. Helliwell, Extraction and quantification of chlorophyll a from
the maximum biomass growth of microalgae was increased by freshwater green algae, Water Res. 32 (7) (1998) 2220–2223, https://doi.org/
38.80% in symbiotic association with a single selected bacterial 10.1016/S0043-1354(97)00452-1.
strain, Pseudomonas aeruginosa.This is is higher than 3.5 to 24.8% [13] X. Zhao et al., Characterization of microalgae-bacteria consortium cultured in
landfill leachate for carbon fixation and lipid production, Bioresour. Technol.
increase of biomass with Scenedesmus obliquus and selected bacte-
156 (2014) 322–328, https://doi.org/10.1016/j.biortech.2013.12.112.
rial strain reported by R. Wang et al. [21]. The biomass productivity [14] J. Shi, P.K. Pandey, A.K. Franz, H. Deng, R. Jeannotte, Chlorella vulgaris
of the consortium was 145.59 mg
1L
1day
1. The maximum COD production enhancement with supplementation of synthetic medium in dairy
manure wastewater, AMB Express 6 (1) (2016) 1–9, https://doi.org/10.1186/
removal by consortia was 87.49% (from 1953.84 to 184.45 mg
s13568-016-0184-1.
L
1). Also, the SM is the best chlorophyll extraction method com- [15] R. Amit, U.K. Chandra, Ghosh, J.K. Nayak, Phycoremediation potential of
pared to the SSM, since the maximum recovery of chlorophyll-a marine microalga Tetraselmis indica on secondary treated domestic sewage
is obtained through SM; also, methanol is a superior extracting sol- for nutrient removal and biodiesel production, Environ. Sci. Pollut. Res. 24 (26)
(2017) 20868–20875, https://doi.org/10.1007/s11356-017-9734-6.
vent for chlorophyll extraction. [16] X. Ji, M. Jiang, J. Zhang, X. Jiang, Z. Zheng, The interactions of algae-bacteria
symbiotic system and its effects on nutrients removal from synthetic
wastewater, Bioresour. Technol. 247 (2018) 44–50, https://doi.org/10.1016/j.
Declaration of Competing Interest biortech.2017.09.074.
[17] M. Amin, P. Chetpattananondh, M.N. Khan, F. Mushtaq, S.K. Sami, Extraction
The authors declare that they have no known competing finan- and Quantification of Chlorophyll from Microalgae Chlorella sp, IOP Conf. Ser.
Mater. Sci. Eng. 414 (1) (2018), https://doi.org/10.1088/1757-899X/414/1/
cial interests or personal relationships that could have appeared
012025.
to influence the work reported in this paper. [18] F. Johan, M. Z. Jafri, H. S. Lim, and W. M. W. O, ‘‘Laboratory measurement :
Chlorophyll-a concentration measurement with acetone method using
spectrophotometer Laboratory Measurement : Chlorophyll-a Concentration
Acknowledgments Measurement with Acetone Method 8Ving Spectrophotometer,” no. March,
2015, doi: 10.1109/IEEM.2014.7058737.
This research work is backed by the Department of Polymer and [19] S. Van Den Hende, H. Vervaeren, H. Saveyn, G. Maes, N. Boon, Microalgal
bacterial floc properties are improved by a balanced inorganic/organic carbon
Process Engineering, IIT Roorkee, India. Financial support to exe- ratio, Biotechnol. Bioeng. 108 (3) (2011) 549–558, https://doi.org/10.1002/
cute this study is gratefully acknowledged to MHRD (Ministry of bit.22985.
Human Resources Development) Plan grant (2020-2021) and IIT [20] J. Sharma et al., Microalgal consortia for municipal wastewater treatment –
Lipid augmentation and fatty acid profiling for biodiesel production, J.
Roorkee (No. OH-35-71-142), IIT Roorkee, India. Photochem. Photobiol. B Biol. 202 (October) (2020), https://doi.org/10.1016/
j.jphotobiol.2019.111638.
[21] R. Wang et al., Construction and characteristics of artificial consortia of
References
Scenedesmus obliquus-bacteria for S. obliquus growth and lipid production,
Algal Res. 12 (2015) 436–445, https://doi.org/10.1016/j.algal.2015.10.002.
[1] I. Priyadarshani, B. Rath, ‘‘Commercial and industrial applications of micro
algae – A review,” vol. 3, no. 4, pp. 89–100, 2012.
[2] J. Lian, R.H. Wijffels, H. Smidt, D. Sipkema, The effect of the algal microbiome Further Reading
on industrial production of microalgae, Microb. Biotechnol. 11 (5) (2018) 806–
818, https://doi.org/10.1111/1751-7915.13296. [1] X. Ji, M. Jiang, J. Zhang, X. Jiang, Z. Zheng, The interactions of algae-bacteria
[3] B. R. Glick, Plant Growth-Promoting Bacteria : Mechanisms and Applications,‘‘ symbiotic system and its effects on nutrients removal from synthetic
vol. 2012, 2012. wastewater, Bioresour. Technol. 247 (September) (2018) 2017, https://doi.
[4] K. Chokshi, I. Pancha, A. Ghosh, S. Mishra, Microalgal biomass generation by org/10.1016/j.biortech.2017.09.074.
phycoremediation of dairy industry wastewater: An integrated approach