Exploring biofilm-forming bacteria for integration into BioCircuit wastewater treatment

Chontisa Sukkasem

doi:10.62063/ecb-28

Authors

Chontisa Sukkasem Department of Food Science and Technology, Faculty of Agro and Bio Industry, Thaksin University, Thailand https://orcid.org/0000-0001-8043-4981

DOI:

https://doi.org/10.62063/ecb-28

Keywords:

BioCircuit system, biofilm-forming bacteria, chemical oxygen demand, sulfate, sulfide, wastewater treatment

Abstract

This study aimed to investigate the presence of biofilm-forming bacteria within high sulfide sludge obtained from a rubber wastewater treatment plant and assess their suitability for application within a BioCircuit System (BCS) as a symbiotic community for treating nutrient-rich wastewater. The sludge samples were collected and subjected to microbial culture techniques, wherein pure cultures were isolated based on morphological characteristics observed under a light microscope, followed by assessment of motility using swarm agar. Subsequent identification was conducted utilizing the 16S rRNA gene sequencing method, and the isolated bacteria were introduced into the BCS. A 12 mL microbial fuel cell test was conducted to evaluate their power generation capabilities. The wastewater treatment process involved inoculating the BCS with 20% crude rubber wastewater sludge, and the system was initiated at a flow rate of 0.5 L/min for a month. Upon achieving an open-circuit voltage exceeding 50 mV, the BCS was operated at incremental flow rates (0.5-1.0, 1.0-1.5, and 1.5-2.0 mL/ min) over a period of 6 months. Real-time monitoring of voltage, flow rate, and energy consumption was facilitated through an internet-of-things online program. Weekly sampling and analysis of influent and effluent, focusing on chemical oxygen demand (COD), sulfate, and sulfide concentrations, were conducted. Additionally, the BioCircuit voltage was recorded every 5 minutes. The results revealed the presence of six group-forming shaped bacteria identified as Bacillus tequilensis, Bacillus sp., Ferribacterium limneticum, Bacillus weihenstephanesis, and Mycobacterium sp., respectively. The optimal flow rate of 1.5 L/min yielded a maximum voltage of 1.2 V and demonstrated high wastewater treatment efficiency. Economically, the BCS operation exhibited a power consumption rate of 0.257 kWhr/m³ of treated wastewater, leading to an 88.90% reduction in carbon footprint compared to aerated lagoon treatment, equivalent to 50.94 kg CO₂/m³ of treated wastewater or 183,384 kg CO₂/yr for a 10 m³ plant. These findings underscore the potential of the BCS in conjunction with group-forming shaped bacteria communities for various industrial wastewater treatment applications.

References

Alvarez, H. M., & Steinbüchel, A. (2002). Physiology, biochemistry, and molecular biology of mycobacteria. Advances in biochemical engineering/biotechnology, 74, 103–140. https://doi.org/10.1007/3-540-45790-0_4

Asensio, Y., Fernandez-Marchante, C. M., Lobato, J., Canizares, P., & Rodrigo, M. A. (2016). Influence of the fuel and dosage on the performance of double-compartment microbial fuel cells. Water research, 99, 16-23. https://doi.org/10.1016/j.watres.2016.04.028

Bridier, A., Briandet, R., Thomas, V., & Dubois-Brissonnet, F. (2011). Resistance of bacterial biofilms to disinfectants: a review. Biofouling, 27(9), 1017-1032. https://doi.org/10.1080/08927014.2011.626899

Davey, M. E., & O’toole, G. A. (2000). Microbial biofilms: from ecology to molecular genetics. Microbiology and molecular biology reviews, 64(4), 847-867. https://doi.org/10.1128/MMBR.64.4.847-867.2000

Fan, Y., Janicek, A., & Liu, H. (2024). Stable and high voltage and power output of CEA-MFCs internally connected in series (iCiS-MFC). The European chemistry and biotechnology journal, (1), 47–57. https://doi.org/10.62063/ecb-17

Cummings, D., Caccavo Jr., F., Spring, S., & Rosenzweig, R.F. (1999). Ferribacterium limneticum, gen. nov., sp. nov., an Fe(III)-reducing microorganism isolated from mining-impacted freshwater lake sediments. Archieves of microbiology, 171, 183–188. https://doi.org/10.1007/s002030050697

Flemming, H. C., & Wingender, J. (2010). The biofilm matrix. Nature reviews microbiology, 8(9), 623-633. https://doi.org/10.1038/nrmicro2415

Hall-Stoodley, L., Costerton, J. W., & Stoodley, P. (2004). Bacterial biofilms: from the natural environment to infectious diseases. Nature reviews microbiology, 2(2), 95-108. https://doi.org/10.1038/nrmicro821

Huang, L., & Logan, B. E. (2008). Electricity generation and treatment of paper recycling wastewater using a microbial fuel cell. Applied microbiology and biotechnology, 80(2), 349-355. https://doi.org/10.1007/s00253-008-1546-7

Kim, J. R., Jung, S. H., Regan, J. M., & Logan, B. E. (2007). Electricity generation and microbial community analysis of alcohol powered microbial fuel cells. Bioresource technology, 98(13), 2568-2577. https://doi.org/10.1016/j.biortech.2006.09.036

Logan, B. E., Hamelers, B., Rozendal, R., Schröder, U., Keller, J., Freguia, S., Aelterman, P., Verstraete, W., & Rabaey, K. (2006). Microbial fuel cells: methodology and technology. Environmental science and technology, 40(17), 5181-5192. https://doi.org/10.1021/es0605016

Logan, N. A., & De Vos, P. (2009). Genus I. Bacillus Cohn 1872. In P. De Vos et al. (Eds.), Bergey’s Manual of Systematic Bacteriology (2nd ed., Vol. 3, pp. 21-127). Springer.

Martin, H. M., & Huq, A. (2007). Bacillus weihenstephanensis Isolated from Fermented Fish Product, Hentak, in Bangladesh. Journal of food protection, 70(4), 1016–1020.

Quan, X. C., Quan, Y. P., & Tao, K. (2012). Effect of anode aeration on the performance and microbial community of an air-cathode microbial fuel cell. Chemical engineering journal, 210, 150-156. https://doi.org/10.1016/j.cej.2012.09.009

Rabaey, K., Boon, N., Höfte, M., & Verstraete, W. (2005). Microbial phenazine production enhances electron transfer in biofuel cells. Environmental science and technology, 39(9), 3401-3408. https://doi.org/10.1021/es048563o

Sonmez, E., Avci, B., Mohamed, N., & Bermek, H. (2024). Investigation of performance losses in microbial fuel cells with low platinum loadings on air-cathodes. The European chemistry and biotechnology journal, (1), 11–26. https://doi.org/10.62063/ecb-14

Stewart, P. S., & Franklin, M. J. (2008). Physiological heterogeneity in biofilms. Nature reviews microbiology, 6(3), 199-210. https://doi.org/10.1038/nrmicro1838

Sukkasem, C., Xu, S., Park, S., Boonsawang, P., & Liu, H. (2008). Effect of nitrate on the performance of single chamber air cathode microbial fuel cells. Water research, 42(19), 4743-4750. https://doi.org/10.1016/j.watres.2008.08.029

Sukkasem, C., Laehlah, S., Nhimman, A., O’thong,S., Boonsawang, P., Rarngnarong, A., Nisoa, M., & Kirdtongmee, P. (2011). Upflow bio-filter circuit (UBFC) biocatalyst microbial fuel cell (MFC) configuration and application to biodiesel wastewater treatment. Bioresource technology, 102, 10363-10370. https://doi.org/10.1016/j.biortech.2011.09.007

Sukkasem, C., & Laehlah, S. (2013). Development of a UBFC biocatalyst fuel cell to generate power and treat industrial wastewaters. Bioresource technology, 146, 749-753. https://doi.org/10.1016/j.biortech.2013.07.065

Sukkasem, C., & Laehlah, S. (2015). An economical upflow bio-filter circuit (UBFC): a biocatalyst microbial fuel cell for sulfate- sulfide rich wastewater treatment. Water research, 1(1), 8. https://doi.org/10.1039/C4EW00028E

Wang, X., Cheng, S. A., Zhang, X. Y., Li, X. Y., & Logan, B. E. (2011). Impact of salinity on cathode catalyst performance in microbial fuel cells (MFCs). International journal of hydrogen energy, 36(21), 13900-13906. https://doi.org/10.1016/j.ijhydene.2011.03.052

Zhuo, R., Ma, L., Fan, F. F., Gong, Y. M., Wan, X., Jiang, M. L., Zhang, X. Y., & Yang, Y. (2011). Decolorization of different dyes by a newly isolated white-rot fungi strain Ganoderma sp.En3 and cloning and functional analysis of its laccase gene. Journal of hazardous materials, 192(2), 855- 873. https://doi.org/10.1016/j.jhazmat.2011.05.106