Evaluation of Biomass Production and Wastewater Nutrient Removal Using Microalgae: Sustainable Strategy to CO2 Bio-Fixation and Bioenergy Production Approach

Document Type: Research Article


1 Department of Renewable Energies and Environmental Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.

2 Department of Biosystems Engineering, University of Tarbiat Modares, Tehran, Iran

3 School of Chemical Engineering, College of Engineering, University of Tehran, Tehran, Iran

4 Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.


Nowadays, the replacement of renewable energies such as biofuels is one of the main priorities in environmental programming and investments. This study is based on sustainable strategy towards integrating algal biomass generation as a green feedstock with wastewater treatment, CO2 bio-fixation, and bioenergy production. Therefore, the performance of Trichormus variabilis in biomass production together with ammonium and phosphate removal from an actual effluent obtained from a mixed wastewater streams has been investigated using two mixing methods under aeration and agitation conditions. Dilutions of 10 %, 20 %, 40 %, 60 %, 90 % and 100 % (v/v) were used for growth evaluation. The results showed that the bubbled air effectively enhances the biomass productivity. However, the agitation system was suggested to cultivate the algae in the wastewater due to the elimination of possible mechanical stimulation stress on cells. Moreover, high pH levels (pH>8) indicated a negative inhibitory effect on growth. Thus, unexpected inhibitory impacts were removed through providing the wastewater dilutions mixed with BG11 culture medium, which contains essential required nutrients, to support the algal growth in the wastewater, adjust pH and remove the mechanical stress induced by bubbling compressed air. The results with respect to investigating the effect of the inoculums and wastewater concentrations on the biomass production suggested that the highest biomass generates with 30 mg.mL-1 inoculum in 40 % mixed wastewater diluted by the BG11 medium having the highest potential in CO2 bio-fixation of 9.19±0.64 g.L-1. The results of the wastewater analysis demonstrated the removal potential of ~43 % and ~75 % for NH4+ and PO43−, respectively. The generated biomass after phycoremediation and CO2 bio-fixation can be effectively utilized in different types of biofuel production.


Main Subjects

1.     Greenwell, H.C., Laurens, L.M.L., Shields, R.J., Lovitt, R.W. and Flynn, K.J., "Placing microalgae on the biofuels priority list: A review of the technological challenges",. Journal of The Royal Society Interface, Vol. 7, No. 46,(2010), 703-726.

2.     Greenland, S.J., Dalrymple, J., Levin, E. and O’Mahony, B., Improving agricultural water sustainability: Strategies for effective farm water management and encouraging the uptake of drip irrigation, in The goals of sustainable development: Responsibility and governance, Crowther, D., Seifi, S. and Moyeen, A. Editors., Springer, Singapore, Singapore,(2018), 111-123.

3.     Connor, R., Faurès, J.M., Kuylenstierna, J., Margat, J., Steduto, P., Vallée, D. and Hoe, W.V.D. , Chapter 7: Evolution of water use., in The United nations world water development report 3–Water in a changing world, E.K. (WWAP) Editor., UNESCO, London.(2009).

4.     Connor, R., Renata, A., Ortigara, C., Koncagül, E., Uhlenbrook, S., Lamizana-Diallo, B.M., Zadeh, S.M., Qadir, M., Kjellén, M. and Sjödin, J., The united nations world water development report 2017, Wastewater: The untapped resource, facts and figures in "WWAP (United Nations World Water Assessment Programme), Paris, UNESCO,(2017).

5.     Ahluwalia, S.S. and Goyal, D., "Microbial and plant derived biomass for removal of heavy metals from wastewater", Bioresource Technology, Vol. 98, No. 12,(2007), 2243-2257.

6.     Markou, G. and Georgakakis, D., "Cultivation of filamentous cyanobacteria (blue-green algae) in agro-industrial wastes and wastewaters: A review", Applied Energy, Vol. 88, No. 10,(2011), 3389-3401.

7.     Larsdotter, K., "Wastewater treatment with microalgae-a literature review", Vatten, Vol. 62, No. 1,(2006), 31.

8.     Razzak, S.A., Hossain, M.M., Lucky, R.A., Bassi, A.S. and. de Lasa, H, "Integrated CO2 capture, wastewater treatment and biofuel production by microalgae culturing: A review", Renewable and Sustainable Energy Reviews, Vol. 27,(2013), 622-653.

9.     Singh, N.K., Upadhyay, A.K. and Rai, U.N., Algal technologies for wastewater treatment and biofuels production: An integrated approach for environmental management, in Algal biofuels: recent advances and future prospects, Gupta, S.K., Malik, A. and Bux, F. Editors., Springer International Publishing: Cham.,(2017), 97-107.

10.   Pittman, J.K., Dean, A.P. and Osundeko, O., "The potential of sustainable algal biofuel production using wastewater resources", Bioresource Technology, Vol. 102, No. 1,(2011), 17-25.

11.   Rawat, I., Ranjith Kumar, R., Mutanda, T. and Bux, F., "Dual role of microalgae: Phycoremediation of domestic wastewater and biomass production for sustainable biofuels production", Applied Energy, Vol. 88, No. 10,(2011), 3411-3424.

12.   Cho, S., Luong, T.T., Lee, D., Oh, Y.-K. and Lee, T., "Reuse of effluent water from a municipal wastewater treatment plant in microalgae cultivation for biofuel production", Bioresource Technology, Vol. 102, No. 18,(2011), 8639-8645.

13.   Maity, J.P., Bundschuh, J., Chen, C.-Y. and Bhattacharya, P., "Microalgae for third generation biofuel production, mitigation of greenhouse gas emissions and wastewater treatment: Present and future perspectives: A mini review", Energy, Vol. 78, No. Supplement C,(2014), 104-113.

14.   Kligerman, D.C. and Bouwer, E.J., "Prospects for biodiesel production from algae-based wastewater treatment in Brazil: A review", Renewable and Sustainable Energy Reviews, Vol. 52,(2015), 1834-1846.

15.   Ruiz-Marin, A., Mendoza-Espinosa, L.G. and. Stephenson, T, "Growth and nutrient removal in free and immobilized green algae in batch and semi-continuous cultures treating real wastewater", Bioresource Technology, Vol. 101, No. 1,(2010), 58-64.

16.   Samorì, G., Samorì, C., Guerrini, F. and Pistocchi, R., "Growth and nitrogen removal capacity of Desmodesmus communis and of a natural microalgae consortium in a batch culture system in view of urban wastewater treatment: Part I", Water Research, Vol. 47, No. 2,(2013), 791-801.

17.   Martı́nez, M.E., Sánchez, S., Jiménez, J.M., El Yousfi, F. and Muñoz, L., "Nitrogen and phosphorus removal from urban wastewater by the microalga Scenedesmus obliquus", Bioresource Technology, Vol. 73, No. 3,(2000), 263-272.

18.   de-Bashan, L.E., Hernandez, J.-P., Morey, T. and Bashan, Y., "Microalgae growth-promoting bacteria as “helpers” for microalgae: A novel approach for removing ammonium and phosphorus from municipal wastewater", Water Research, Vol. 38, No. 2,(2004), 466-474.

19.   El-Sheekh, M.M., El-Shouny, W.A., Osman, M.E.H. and El-Gammal, E.W.E., "Growth and heavy metals removal efficiency of Nostoc muscorum and Anabaena subcylindrica in sewage and industrial wastewater effluents", Environmental Toxicology and Pharmacology, Vol. 19, No. 2,(2005), 357-365.

20.   González, L.E., Cañizares, R.O. and Baena, S., "Efficiency of ammonia and phosphorus removal from a colombian agroindustrial wastewater by the microalgae Chlorella vulgaris and Scenedesmus dimorphus", Bioresource Technology, Vol. 60, No. 3,(1997), 259-262.

21.   Singh, N.K. and Dhar, D.W., "Microalgal remediation of sewage effluent", Proceedings of The Indian National Science Academy, Vol. 76, No. 4,(2010), 209-221.

22.   Aslan, S. and Kapdan, I.K., "Batch kinetics of nitrogen and phosphorus removal from synthetic wastewater by algae", Ecological Engineering, Vol. 28, No. 1,(2006), 64-70.

23.   Kim, T.-H., Lee, Y., Han, S.-H. and Hwang, S.-J., "The effects of wavelength and wavelength mixing ratios on microalgae growth and nitrogen, phosphorus removal using Scenedesmus sp. for wastewater treatment", Bioresource Technology, Vol. 130, No. Supplement C,(2013), 75-80.

24.   Avery, S.V., Codd, G.A. and Gadd, G.M., Microalgal removal of organic and inorganic metal species from aqueous solution, in Wastewater treatment with algae, Wong, Y.-S. and Tam, N.F.Y. Editors., Springer Berlin Heidelberg, Berlin, Heidelberg, (1998), 55-72.

25.   Prince, I.G., Ting, Y.P. and Lawson, F., Modeling the uptake of metal ions by living algal cells, in Wastewater treatment with algae, Wong, Y.-S. and Tam, N.F.Y. Editors., Springer Berlin Heidelberg, Berlin, Heidelberg, (1998), 125-144.

26.   Fiore, M.F., Moon, D.H. and Trevors, J.T., Metal resistance and accumulation in cyanobacteria, in Wastewater treatment with algae, Wong, Y.-S. and Tam, N.F.Y. Editors., Springer Berlin Heidelberg, Berlin, Heidelberg, (1998), 111-124.

27.   Aksu, Z., Biosorption of heavy metals by microalgae in match and continuous systems, in Wastewater treatment with algae, Wong, Y.-S. and Tam, N.F.Y. Editors., Springer Berlin Heidelberg, Berlin, Heidelberg, (1998), 37-53.

28.   Zhou, J.L., Huang, P.L. and Lin, R.G., "Sorption and desorption of Cu and Cd by macroalgae and microalgae", Environmental Pollution, Vol. 101, No. 1,(1998), 67-75.

29.   Worku, A. and Sahu, O., "Reduction of heavy metal and hardness from ground water by algae", Journal of Applied & Environmental Microbiology, Vol. 2, No. 3,(2014), 86-89.

30.   Chevalier, P., Proulx, D., Lessard, P., Vincent, W.F. and de la Noüe, J., "Nitrogen and phosphorus removal by high latitude mat-forming cyanobacteria for potential use in tertiary wastewater treatment", Journal of Applied Phycology, Vol. 12, No. 2,(2000), 105-112.

31.   El-Bestawy, E., "Treatment of mixed domestic-industrial wastewater using cyanobacteria", Journal of Industrial Microbiology and Biotechnology, Vol. 35, No. 11,(2008), 1503-16.

32.   Fawzy, M.A. and Issa, A.A., "Bioremoval of heavy metals and nutrients from sewage plant by Anabaena oryzae and Cyanosarcina fontana", International Journal of Phytoremediation, Vol. 18, No. 4,(2016), 321-328.

33.   Borowitzka, M.A., Limits to Growth, in Wastewater treatment with algae, Wong, Y.-S. and Tam, N.F.Y. Editors., Springer Berlin Heidelberg, Berlin, Heidelberg, (1998), 203-226.

34.   Komarek, J.a.K., Anagnostidis, "Modern approach to the classification system of Cyanophytes 4-Nostocales", Arch. Hydrobiol. Suppl. 82(3), Algological Studies/Archiv für Hydrobiologie, Supplement Volumes, Vol. 56,(1989), 247-345.

35.   Han, F., Pei, H., Hu, W., Jiang, L., Cheng, J. and Zhang, L., "Beneficial changes in biomass and lipid of microalgae Anabaena variabilis facing the ultrasonic stress environment", Bioresource Technology, Vol. 209,(2016), 16-22.

36.   Pyo, D., Kim, T. and Yoo, J., "Efficient extraction of bioethanol from freshwater cyanobacteria using supercritical fluid pretreatment", Bull Korean Chem Soc, Vol. 34,(2013), 379-383.

37.   Markov, S.A., Protasov, E.S., Bybin, V.A., Eivazova, E.R. and Stom, D.I., "Using immobilized cyanobacteria and culture medium contaminated with ammonium for H2 production in a hollow-fiber photobioreactor", International Journal of Hydrogen Energy, Vol. 40, No. 14,(2015), 4752-4757.

38.   Petrova, E., Egorova, M., Piskunkova, N., Kozhevin, P., Netrusov, A. and Tsavkelova, E., "Anaerobic cellulolytic microbial communities decomposing the biomass of Anabaena variabilis", Microbiology, Vol. 86, No. 6,(2017), 745-752.

39.   Jia, H. and Yuan, Q., "Removal of nitrogen from wastewater using microalgae and microalgae–bacteria consortia", Cogent Environmental Science, Vol. 2, No. 1,(2016), 1275089.

40.   Parameswari, E., Lakshmanan, A. and Thilagavathi, T., "Phycoremediation of heavy metals in polluted water bodies", Electronic Journal of Environmental, Agricultural and Food Chemistry, Vol. 9, No. 4,(2010), 808-814.

41.   Hirooka, T., Akiyama, Y., Tsuji, N., Nakamura, T., Nagase, H., Hirata, K. and Miyamoto, K., "Removal of hazardous phenols by microalgae under photoautotrophic conditions", Journal of Bioscience and Bioengineering, Vol. 95, No. 2,(2003), 200-203.

42.   Noel, S.D. and Rajan, M., "Cyanobacteria as a potential source of phycoremediation from textile industry effluent", Journal of Microbiology and Biotechnology Research, Vol. 4, No. 6,(2014), 35-33.

43.   Stanier, R., Kunisawa, R., Mandel, M. and Cohen-Bazire, G., "Purification and properties of unicellular blue-green algae (order Chroococcales)", Bacteriological Reviews, Vol. 35, No. 2,(1971), 171.

44.   Yoon, J.H., Shin, J.-H. and Park, T.H., "Characterization of factors influencing the growth of Anabaena variabilis in a bubble column reactor", Bioresource Technology, Vol. 99, No. 5,(2008), 1204-1210.

45.   Ratledge, C. and Kristiansen, B., Basic biotechnology, Cambridge University Press,(2006).

46.   Adamczyk, M., Lasek, J., and Skawińska, A., "CO2 biofixation and growth kinetics of Chlorella vulgaris and Nannochloropsis gaditana", Applied Biochemistry and Biotechnology, Vol. 179, No. 7,(2016), 1248-1261.

47.   Tarin, N., Ali, N., Chamon, A., Mondol, M., Rahman, M. and Aziz, A., "Optimizing Chlorella vulgaris and Anabaena variabilis growth conditions for use as biofule feedstock".

48.   Ronda, S.R., Bokka, C.S., Ketineni, C., Rijal, B. and Allu, P.R., "Aeration effect on Spirulina platensis growth and γ-linolenic acid production", Brazilian Journal of Microbiology, Vol. 43, No. 1,(2012), 12-20.

49.   Berberoglu, H., Barra, N., Pilon, L. and Jay, J., "Growth, CO2 consumption and H2 production of Anabaena variabilis ATCC 29413-U under different irradiances and CO2 concentrations", J. Appl. Microbiol., Vol. 104, No. 1,(2008), 105-121.

50.   DM, T. and T. S., "Phytoremediation– Aeration system for treatment of urban wastewater", Austin Chemial Engineering, Vol. 4, No. 1,(2017), 1047.

51.   Whitton, R., Ometto, F., Pidou, M., Jarvis, P., Villa, R. and. Jefferson, B, "Microalgae for municipal wastewater nutrient remediation: mechanisms, reactors and outlook for tertiary treatment", Environmental Technology Reviews, Vol. 4, No. 1,(2015), 133-148.

52.   Rodrigues, M.N., "Nutrient removal using microalgae in wastewater-fed high rate ponds",(2013).

53.   Bunce, J.T., Ndam, E., Ofiteru, I.D., Moore, A. and Graham, D.W., "A review of phosphorus removal technologies and their applicability to small-scale domestic wastewater treatment systems", Frontiers in Environmental Science, Vol. 6,(2018), 8.

54.   Vargas, M., Rodriguez, H., Moreno, J., Olivares, H., Del Campo, J., Rivas, J. and Guerrero, M., "Biochemical composition and fatty acid content of filamentous nitrogen‐fixing cyanobacteria", Journal of Phycology, Vol. 34, No. 5,(1998), 812-817.

55.   Pyo, D.J., Method for producing ethanol using freshwater blue-green algae in World intellectual property organization (WIPO). WO2010095895A9, Foundation Kangwon National University and Tjd Co., Ltd., .Korea.(2010).

56.   Gassanova, L., Netrusov, A.I., Teplyakov, V. and Modigell, M., "Fuel gases from organic wastes using membrane bioreactors", Desalination, Vol. 198, No. 1-3,(2006), 56-66.

57.   Salleh, S.F., Kamaruddin, A., Uzir, M.H. and Mohamed, A.R., "Effects of cell density, carbon dioxide and molybdenum concentration on biohydrogen production by Anabaena variabilis ATCC 29413", Energy Conversion and Management, Vol. 87,(2014), 599-605.

58.   Berberoğlu, H., Jay, J. and Pilon, L., "Effect of nutrient media on photobiological hydrogen production by Anabaena variabilis ATCC 29413", International Journal of Hydrogen Energy, Vol. 33, No. 4,(2008), 1172-1184.

59.   Yoon, J.H., Sim, S.J., Kim, M.S. and Park, T.H., "High cell density culture of Anabaena variabilis using repeated injections of carbon dioxide for the production of hydrogen", International Journal of Hydrogen Energy, Vol. 27, No. 11-12,(2002), 1265-1270.

60.   Famiglietti, M., Hochkoeppler, A. and Luisi, P.L., "Surfactant-Induced hydrogen production in cyanobacteria", Biotechnology and Bioengineering, Vol. 42, No. 8,(1993), 1014-8.