Microbial Fuel Cell for Bioremediation and Bioenergy Production Using Mixed Bacterial Culture Isolated from Municipal Waste
Abstract
Introduction: Bioelectrical devices are being studied in a pilot project to treat wastewater and provide electricity, focusing on four bacterial strains and their impact on system parameters. Objectives: In this study, four electrogenic microorganisms tolerant to sewage discharge were isolated and evaluated and named AKS2, AKS14, BKS2 and CKW5. Results: Sewage wastewater was treated with these four bacterial strains consortia in different configurations of microbial fuel cells (MFC). Compared to BKS2, which produced a potential difference of 1.916±0.045 V and 7.222±0.051 mA, AKS2 had a higher potential to generate energy, measuring 1.943±0.064 V and 7.793±0.007 mA. On the other hand, CKW5 may produce a potential difference of 7.205±0.039 mA and 1.895±0.066 V, which is higher than the isolates but less than that of AKS2 and BKS2. In pH 7 and 35ºC temperature with 15% (v/v) bacterial inoculum, AKS13 showed higher potential than AKS14, which generated 1.875±0.039 V and 7.195±0.027 mA. Whereas AKS14 generates 1.871±0.006 V and 7.192±0.009 mA. These isolates have the following bioremediation capacities: AKS2 (89.88%) > CKW5 (89%), >BKS2 (88%) > AKS14 (85.48%). Graph Pad PRISM software, version 9.1.5, was used to statistically validate all of the bio remedial percentages using a two-way repeated measure (RM) ANOVA. With a R squared value of 0.1017, the results were determined to be statistically significant at p < 0.01. The study characterized four potent bacterial isolates using 16S rDNA sequencing of AKS2, AKS14, BKS2, and CKW5, revealing Escherichia coli, Salmonella enterica, Bacillus cereus, and Klebsiella pneumonia respectively. The novelty of the present study lies in the systematic isolation, comparative evaluation, and electrical stacking of indigenous electrogenic bacteria from municipal sewage wastewater for simultaneous bioremediation and bioelectricity generation in microbial fuel cells (MFC).
Keywords:
Bioelectricity, COD Removal, Electrogenic Microorganisms, Microbial Fuel Cell, Sewage Wastewater, Salt Bridge Enrichment
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References
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Apollon, W., Rusyn, I., Paucar, N. E., Hibbert, M., Kamaraj, S.-K., & Sato, C. (2025). Energy Recovery from Organic Wastes Using Microbial Fuel Cells: Traditional and Nonconventional Organic Substrates. Resources, 14(3), 47. https://doi.org/10.3390/resources14030047
Ardakani, M. N., and Gholikandi, B. G. (2020) Microbial fuel cells (MFC) in integration with anaerobic treatment processes (antps) and membrane bioreactors (mbrs) for simultaneous efficient wastewater/sludge treatment and energy recovery -A state-of-the-art review. Biomass and Bioenergy, 141, 105726. https://doi.org/10.1016/j.biombioe.2020.105726
Bethoux, O. (2020). Hydrogen fuel cell road vehicles: state of the art and perspectives. Energies, 13(21), 5843. https://doi.org/10.3390/en13215843
Bikram, J. S., Chakraborty, A., & Sehgal, R. (2023). A systematic review of industrial wastewater management: Evaluating challenges and enablers. Journal of Environmental Management, 348(15), 119230. https://doi.org/10.1016/j.jenvman.2023.119230
Bose, D., Bhattacharya, R., Gopinath, M., Sarkar, A., Pandya, R. S., and Jaiswal, A. (2025) Advances in microbial fuel cell technologies for bioremediation and energy recovery from wastewater. Sustainable Chemistry for the Environment, 11, 100266. https://doi.org/10.1016/j.scenv.2025.100266
Campo, G. D. A., Cañizares. P., Rodrigo, M. A., Fernández, F. J., and Lobato, J. (2015) Microbial fuel cell with an algae-assisted cathode: A preliminary assessment. Journal of Power Sources, 242, 638–645. http://dx.doi.org/10.1016/j.jpowsour.2013.05.110.
Chaibi Y, Allouhi A, Malvoni M, Salhi M, Saadani, R. (2019) Solar irradiance and temperature influence on the photovoltaic cell equivalent-circuit models. Solar Energy, 188, 1102-1110. https://doi.org/10.1016/j.solener.2019.07.005
Dakal, T. C., Singh, N., Kaur, A., Dhillon. P. K., Bhatankar, J., Meena, R., Sharma, R. K., Gadi, B. R. Sahu, B. S. Patel, A., Singh, B. and Kumari, K. (2025) New horizons in microbial fuel cell technology: applications, challenges, and prospects. Biotechnology for Biofuels and Bioproducts,18;18:79. 10.1186/s13068-025-02649-y
Du, Z., Li, H., & Gu, T. (2007). A state of the art review on microbial fuel cells: A promising technology for wastewater treatment and bioenergy. Research review paper, Biotechnology Advances, 25(5), 464–482. https://doi.org/10.1016/j.biotechadv.2007.05.004
Elakkiya, E., and Matheswaran, M. (2013). Comparison of anodic metabolisms in bioelectricity production during treatment of dairy wastewater in Microbial Fuel Cell. Bioresource Technology, 136, 407-412. https://doi.org/10.1016/j.biortech.2013.02.113
Fida, S., Yasmeen, M., Adnan, R., & Muhammad, Z. (2025). Treatment methods for sugar rich wastewater: A review. Cleaner Water, 3, 100067. https://doi.org/10.1016/j.clwat.2025.100067
Huang, L., Liu, Y., Yu, L., Quan, X., and Chen, G. (2015). A new clean approach for production of cobalt dihydroxide from aqueous Co(II) using oxygen-reducing biocathode microbial fuel cells. Journal of Cleaner Production, 86, 441–446. https://doi.org/10.1016/j.jclepro.2014.08.018
Jadhav, D. A., Jain, S. C., Ghangrekar, M. M. (2016). Cow’s urine as a yellow gold for bioelectricity generation in low cost clayware microbial fuel cell. Energy, 113(C), 76-84. https://doi.org/10.1016/j.energy.2016.07.025
Larrosa-Guerrero, A., Scott, K., Head, I.M., Mateo, F., Ginesta, A., and Godinez, C. (2010). Effect of temperature on the performance of microbial fuel cells. Fuel, 89 (12) 3985-3994. https://doi.org/10.1016/j.fuel.2010.06.025
Logan, B.E., and Regan, J.M. (2006). Microbial fuel cells: challenges and applications. Environ. Sci. Technol., 40(17):5172–5180. https://doi.org/10.1021/es0627592
Luo, S., Adam, D., Giaveri, S., Barthel, S., Cestellos-Blanco S., Hege, D., Paczia, N., Castan˜ eda-Losada, L., Klose, M., Arndt, F., Heider, J., and Erb, T.J. (2023). ATP production from electricity with a new-to-nature electrobiological module. Joule, 7(8), 1745–1758. https://doi.org/10.1016/j.joule.2023.07.012
Mohana, S., Acharya, B. K., & Madamwar, D. (2009). Distillery spent wash: Treatment technologies and potential applications. Journal of Hazardous Materials, 163(1), 12-25. https://doi.org/10.1016/j.jhazmat.2008.06.079
Nimje, V. R., Chen, C. Y., Chen, H. R., Chen, C. C., Huang, Y. M., Tseng, M. J., Cheng, K. C., & Chang, Y. F. (2012). Comparative bioelectricity production from various wastewaters in microbial fuel cells using mixed cultures and a pure strain of Shewanella oneidensis. Bioresour. Technology, 104, 315–323. https://doi.org/10.1016/j.biortech.2011.09.129
Obileke, K.C., Onyeaka, H., Meyer, E.L., & Nwokolo, N. (2021). Microbial fuel cells, a renewable energy technology for bio-electricity generation: A mini review. Electrochemistry Communications, 125, 107003. https://doi.org/10.1016/j.elecom.2021.107003
Pal, A. and Paul, A. K. (2008) Microbial extracellular polymeric substances: central elements in heavy metal bioremediation. Indian Journal of Microbiology 48(1): 49–64. https://doi.org/10.1007/s12088-008-0006-5
Pant, D., and Adholeya, A. (2007) Biological approaches for treatment of distillery wastewater. A review. Bioresource Technology, 98(12), 2321–2334. https://doi.org/10.1016/j.biortech.2006.09.027
Pant, D., Bogaert V. G., Diels, L., & Vanbroekhoven, K. (2010). A Review of the Substrates Used in Microbial Fuel Cells (MFC) for Sustainable Energy Production. Bioresource Technology, 101(6), 1533-1543. https://doi.org/10.1016/j.biortech.2009.10.017
Prakash, O., Mungray, A., Chongdar, S., Mungray, A. K., and Kailasa, S. K. (2018) Performance of polypyrrole coated metal oxide composite electrodes for benthic microbial fuel cell (BMFC). Journal of Environmental Chemical Engineering 8(2): 102757. https://doi.org/10.1016/j.jece.2018.11.002
Raghavulu, S. V., Modestra, J. A., Amulya, K., Reddy, C. N., Venkata Mohan, S. (2013). Relative effect of bioaugmentation with electrochemically active and non-active bacteria on bioelectrogenesis in microbial fuel cell. Bioresour. Technol. 146, 696–703. https://doi.org/10.1016/j.biortech.2013.07.097
Rodrigues, R., Du, Y., Antoniazzi, A., and Cairoli, P. (2021). A Review of Solid-State Circuit Breakers. IEEE Transactions on Power Electronics, 36(1), 364-377 https://doi.org/10.1109/TPEL.2020.3003358
Saha, N. K., Balakrishnan, M., & Batra, V. S. (2005) Improving industrial water use: case study for an Indian distillery. Resources, Conservation and Recycling. 43(2), 163-174. https://doi.org/10.1016/j.resconrec.2004.04.016
Saxena, A., Gupta, V., and Saxena, S. (2023) Identification and characterization of Microbial Consortia present in sewage samples collected from Sewage Treatment Plant, Jaipur, (Raj.). J. Integr. Sci. Technol. 11(3), 516.
Saxena, A., Gupta, V., and Saxena, S. (2024) Effect of pH and Temperature on Mediator-Less Microbial Fuel Cells for Wastewater Degradation and Sustainable Bioenergy Production, J. Integr. Sci. Technol. 12(2), 726.
Selmi, T., Khadhraoui, A., and Cherif, A. (2022) Fuel cell–based electric vehicles technologies and challenges. Environ Sci Pollut Res 29, 78121–78131. https://doi.org/10.1007/s11356-022-23171-w
Sonawane, A.V., Rikame. S., Sonawane, S.H., Gaikwad, M., Bhanvase, B., Sonawane, S.S., and Gaikwad, R. (2024) A review of microbial fuel cell and its diversification in the development of green energy technology. Chemosphere. 350,141127. https://doi.org/10.1016/j.chemosphere.2024.141127
Sutar, A. A., Dashpute, R. S., Shinde, Y. D., Mukherjee, S., and Chowdhury, C. (2024) A Systemic Review on Fitness and Survival of Salmonella in Dynamic Environment and Conceivable Ways of Its Mitigation. Indian J Microbiol. https://doi.org/10.1007/s12088-023-01176-4
Tewari, P. K., Batra, V. S., & Balakrishnan, M. (2007). Water management initiatives in sugarcane molasses based distilleries in India. Resources, Conservation and Recycling, 52(2), 351-367. https://doi.org/10.1016/j.resconrec.2007.05.003
Venkata Mohan, S., Mohanakrishna, G., Velvizhi, G., Babu, V. L., & Sarma, P. N. (2010). Bio-catalyzed electrochemical treatment of real field dairy wastewater with simultaneous power generation. Biochemical Engineering Journal, 51(1-2), 32–39. https://doi.org/10.1016/j.bej.2010.04.012
Wen, Q., Wu, Y., Zhao, L., Sun, Q., & Kong, F. (2010). Electricity generation and brewery wastewater treatment from sequential anode-cathode microbial fuel cell. Journal of Zhejiang University SCIENCE B, 11, 87–93. https://doi.org/10.1631/jzus.B0900272
Agrahari, R., Bayar, B., Abubackar, A. N., Giri, B. S., Rene, E. R., and Rani, R. (2022). Advances in the development of electrode materials for improving the reactor kinetics in microbial fuel cells. Chemosphere, 290, 133184(2022), https://doi.org/10.1016/j.chemosphere.2021.133184
Apollon, W., Rusyn, I., Paucar, N. E., Hibbert, M., Kamaraj, S.-K., & Sato, C. (2025). Energy Recovery from Organic Wastes Using Microbial Fuel Cells: Traditional and Nonconventional Organic Substrates. Resources, 14(3), 47. https://doi.org/10.3390/resources14030047
Ardakani, M. N., and Gholikandi, B. G. (2020) Microbial fuel cells (MFC) in integration with anaerobic treatment processes (antps) and membrane bioreactors (mbrs) for simultaneous efficient wastewater/sludge treatment and energy recovery -A state-of-the-art review. Biomass and Bioenergy, 141, 105726. https://doi.org/10.1016/j.biombioe.2020.105726
Bethoux, O. (2020). Hydrogen fuel cell road vehicles: state of the art and perspectives. Energies, 13(21), 5843. https://doi.org/10.3390/en13215843
Bikram, J. S., Chakraborty, A., & Sehgal, R. (2023). A systematic review of industrial wastewater management: Evaluating challenges and enablers. Journal of Environmental Management, 348(15), 119230. https://doi.org/10.1016/j.jenvman.2023.119230
Bose, D., Bhattacharya, R., Gopinath, M., Sarkar, A., Pandya, R. S., and Jaiswal, A. (2025) Advances in microbial fuel cell technologies for bioremediation and energy recovery from wastewater. Sustainable Chemistry for the Environment, 11, 100266. https://doi.org/10.1016/j.scenv.2025.100266
Campo, G. D. A., Cañizares. P., Rodrigo, M. A., Fernández, F. J., and Lobato, J. (2015) Microbial fuel cell with an algae-assisted cathode: A preliminary assessment. Journal of Power Sources, 242, 638–645. http://dx.doi.org/10.1016/j.jpowsour.2013.05.110.
Chaibi Y, Allouhi A, Malvoni M, Salhi M, Saadani, R. (2019) Solar irradiance and temperature influence on the photovoltaic cell equivalent-circuit models. Solar Energy, 188, 1102-1110. https://doi.org/10.1016/j.solener.2019.07.005
Dakal, T. C., Singh, N., Kaur, A., Dhillon. P. K., Bhatankar, J., Meena, R., Sharma, R. K., Gadi, B. R. Sahu, B. S. Patel, A., Singh, B. and Kumari, K. (2025) New horizons in microbial fuel cell technology: applications, challenges, and prospects. Biotechnology for Biofuels and Bioproducts,18;18:79. 10.1186/s13068-025-02649-y
Du, Z., Li, H., & Gu, T. (2007). A state of the art review on microbial fuel cells: A promising technology for wastewater treatment and bioenergy. Research review paper, Biotechnology Advances, 25(5), 464–482. https://doi.org/10.1016/j.biotechadv.2007.05.004
Elakkiya, E., and Matheswaran, M. (2013). Comparison of anodic metabolisms in bioelectricity production during treatment of dairy wastewater in Microbial Fuel Cell. Bioresource Technology, 136, 407-412. https://doi.org/10.1016/j.biortech.2013.02.113
Fida, S., Yasmeen, M., Adnan, R., & Muhammad, Z. (2025). Treatment methods for sugar rich wastewater: A review. Cleaner Water, 3, 100067. https://doi.org/10.1016/j.clwat.2025.100067
Huang, L., Liu, Y., Yu, L., Quan, X., and Chen, G. (2015). A new clean approach for production of cobalt dihydroxide from aqueous Co(II) using oxygen-reducing biocathode microbial fuel cells. Journal of Cleaner Production, 86, 441–446. https://doi.org/10.1016/j.jclepro.2014.08.018
Jadhav, D. A., Jain, S. C., Ghangrekar, M. M. (2016). Cow’s urine as a yellow gold for bioelectricity generation in low cost clayware microbial fuel cell. Energy, 113(C), 76-84. https://doi.org/10.1016/j.energy.2016.07.025
Larrosa-Guerrero, A., Scott, K., Head, I.M., Mateo, F., Ginesta, A., and Godinez, C. (2010). Effect of temperature on the performance of microbial fuel cells. Fuel, 89 (12) 3985-3994. https://doi.org/10.1016/j.fuel.2010.06.025
Logan, B.E., and Regan, J.M. (2006). Microbial fuel cells: challenges and applications. Environ. Sci. Technol., 40(17):5172–5180. https://doi.org/10.1021/es0627592
Luo, S., Adam, D., Giaveri, S., Barthel, S., Cestellos-Blanco S., Hege, D., Paczia, N., Castan˜ eda-Losada, L., Klose, M., Arndt, F., Heider, J., and Erb, T.J. (2023). ATP production from electricity with a new-to-nature electrobiological module. Joule, 7(8), 1745–1758. https://doi.org/10.1016/j.joule.2023.07.012
Mohana, S., Acharya, B. K., & Madamwar, D. (2009). Distillery spent wash: Treatment technologies and potential applications. Journal of Hazardous Materials, 163(1), 12-25. https://doi.org/10.1016/j.jhazmat.2008.06.079
Nimje, V. R., Chen, C. Y., Chen, H. R., Chen, C. C., Huang, Y. M., Tseng, M. J., Cheng, K. C., & Chang, Y. F. (2012). Comparative bioelectricity production from various wastewaters in microbial fuel cells using mixed cultures and a pure strain of Shewanella oneidensis. Bioresour. Technology, 104, 315–323. https://doi.org/10.1016/j.biortech.2011.09.129
Obileke, K.C., Onyeaka, H., Meyer, E.L., & Nwokolo, N. (2021). Microbial fuel cells, a renewable energy technology for bio-electricity generation: A mini review. Electrochemistry Communications, 125, 107003. https://doi.org/10.1016/j.elecom.2021.107003
Pal, A. and Paul, A. K. (2008) Microbial extracellular polymeric substances: central elements in heavy metal bioremediation. Indian Journal of Microbiology 48(1): 49–64. https://doi.org/10.1007/s12088-008-0006-5
Pant, D., and Adholeya, A. (2007) Biological approaches for treatment of distillery wastewater. A review. Bioresource Technology, 98(12), 2321–2334. https://doi.org/10.1016/j.biortech.2006.09.027
Pant, D., Bogaert V. G., Diels, L., & Vanbroekhoven, K. (2010). A Review of the Substrates Used in Microbial Fuel Cells (MFC) for Sustainable Energy Production. Bioresource Technology, 101(6), 1533-1543. https://doi.org/10.1016/j.biortech.2009.10.017
Prakash, O., Mungray, A., Chongdar, S., Mungray, A. K., and Kailasa, S. K. (2018) Performance of polypyrrole coated metal oxide composite electrodes for benthic microbial fuel cell (BMFC). Journal of Environmental Chemical Engineering 8(2): 102757. https://doi.org/10.1016/j.jece.2018.11.002
Raghavulu, S. V., Modestra, J. A., Amulya, K., Reddy, C. N., Venkata Mohan, S. (2013). Relative effect of bioaugmentation with electrochemically active and non-active bacteria on bioelectrogenesis in microbial fuel cell. Bioresour. Technol. 146, 696–703. https://doi.org/10.1016/j.biortech.2013.07.097
Rodrigues, R., Du, Y., Antoniazzi, A., and Cairoli, P. (2021). A Review of Solid-State Circuit Breakers. IEEE Transactions on Power Electronics, 36(1), 364-377 https://doi.org/10.1109/TPEL.2020.3003358
Saha, N. K., Balakrishnan, M., & Batra, V. S. (2005) Improving industrial water use: case study for an Indian distillery. Resources, Conservation and Recycling. 43(2), 163-174. https://doi.org/10.1016/j.resconrec.2004.04.016
Saxena, A., Gupta, V., and Saxena, S. (2023) Identification and characterization of Microbial Consortia present in sewage samples collected from Sewage Treatment Plant, Jaipur, (Raj.). J. Integr. Sci. Technol. 11(3), 516.
Saxena, A., Gupta, V., and Saxena, S. (2024) Effect of pH and Temperature on Mediator-Less Microbial Fuel Cells for Wastewater Degradation and Sustainable Bioenergy Production, J. Integr. Sci. Technol. 12(2), 726.
Selmi, T., Khadhraoui, A., and Cherif, A. (2022) Fuel cell–based electric vehicles technologies and challenges. Environ Sci Pollut Res 29, 78121–78131. https://doi.org/10.1007/s11356-022-23171-w
Sonawane, A.V., Rikame. S., Sonawane, S.H., Gaikwad, M., Bhanvase, B., Sonawane, S.S., and Gaikwad, R. (2024) A review of microbial fuel cell and its diversification in the development of green energy technology. Chemosphere. 350,141127. https://doi.org/10.1016/j.chemosphere.2024.141127
Sutar, A. A., Dashpute, R. S., Shinde, Y. D., Mukherjee, S., and Chowdhury, C. (2024) A Systemic Review on Fitness and Survival of Salmonella in Dynamic Environment and Conceivable Ways of Its Mitigation. Indian J Microbiol. https://doi.org/10.1007/s12088-023-01176-4
Tewari, P. K., Batra, V. S., & Balakrishnan, M. (2007). Water management initiatives in sugarcane molasses based distilleries in India. Resources, Conservation and Recycling, 52(2), 351-367. https://doi.org/10.1016/j.resconrec.2007.05.003
Venkata Mohan, S., Mohanakrishna, G., Velvizhi, G., Babu, V. L., & Sarma, P. N. (2010). Bio-catalyzed electrochemical treatment of real field dairy wastewater with simultaneous power generation. Biochemical Engineering Journal, 51(1-2), 32–39. https://doi.org/10.1016/j.bej.2010.04.012
Wen, Q., Wu, Y., Zhao, L., Sun, Q., & Kong, F. (2010). Electricity generation and brewery wastewater treatment from sequential anode-cathode microbial fuel cell. Journal of Zhejiang University SCIENCE B, 11, 87–93. https://doi.org/10.1631/jzus.B0900272
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Saxena, A. and Gupta, V. (2026) “Microbial Fuel Cell for Bioremediation and Bioenergy Production Using Mixed Bacterial Culture Isolated from Municipal Waste”, International Journal of Advancement in Life Sciences Research, 9(2), pp. 42-52. doi: https://doi.org/10.31632/ijalsr.2026.v09i02.004.
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