Screen of microbial strains to reduce H₂S and CH₄ emission from wastewater

Abstract

Reducing H₂S and CH₄ emissions from wastewater is essential for environmental protection and public health. This study screened microbial strains capable of effectively reducing these emissions from anaerobic wastewater and explored their mechanisms. The results demonstrated that Bacillus subtilis and Saccharomyces cerevisiae reduced H₂S emissions by 42.67% and 68.72%, and CH₄ emissions by 58.17% and 77.12%, respectively. Both strains notably elevated the oxidation-reduction potential, which impeded electron transfer efficiency and promoted the accumulation of intermediate sulfur compounds (e.g., S⁰ and S₂O₃²⁻). Furthermore, the presence of B. subtilis and S. cerevisiae also increased the concentrations of longer-chain volatile fatty acids (VFAs), including butyrate and propionate, which are less readily utilized utilization by methanogens, thus contributing to reduced CH₄ production. Microbial community analysis revealed that the relative abundance of Desulfobacterota decreased drastically, by 99.06% in the B. subtilis group and 97.97% in the S. cerevisiae group compared to the control. Additionally, genera associated with the production of lactic acid, propionate, and butyrate became more dominant, thereby limiting the availability of substrates for methanogens and further reducing CH₄ emissions. Functional prediction analyses corroborated these findings, indicating a reduced abundance of genes involved in sulfate reduction and methanogenesis. Overall, this study demonstrates that S. cerevisiae and B. subtilis effectively reduce H₂S and CH₄ emissions by altering microbial community composition and metabolic pathways, offering a sustainable approach for wastewater treatment.

Graphical abstract

Keywords

Wastewater / Sulfide / Methane / Yeast / Microbial agent

Highlight

	● B. subtilis and S. cerevisiae reduced H2S and CH4 emissions in wastewater.
	● Introduced strains increased sulfate reduction intermediates, reduced S2−.
	● Introduced strains inhibited CH4 by varying the concentration and species of VFAs.
	● Introduced strains affected the microbial community structure.
	● Introduced strains reduced functional gene abundance for H2S and CH4 production.

References

[1] Bayrock D, Ingledew W M. (2001). Changes in steady state on introduction of a Lactobacillus contaminant to a continuous culture ethanol fermentation. Journal of Industrial Microbiology & Biotechnology, 27(1): 39–45

[2] Chen D Z, Qiu J, Sun H, Liu Y, Ye J, Chen J M, Lu L. (2024). Enhanced chlorobenzene removal by internal magnetic field through initial cell adhesion and biofilm formation. Applied Microbiology and Biotechnology, 108(1): 159

[3] Dao T T M, Truong D D, Duong L N H, Nguyen N N Y, Nguyen H D. (2023). Preparation of Bacillus subtilis cell samples and generation of an SDS-PAGE. BioTechniques, 74(3): 123–129

[4] Fukuda N. (2023). Apparent diameter and cell density of yeast strains with different ploidy. Scientific Reports, 13(1): 1513

[5] Gutiérrez P, Roldán A, Caro I, Pérez L. (2010). Kinetic study of the velum formation by Saccharomyces cerevisiae (Beticus ssp.) during the biological aging of wines. Process Biochemistry, 45(4): 493–499

[6] Jayaraman A, Mansfeld F B, Wood T K. (1999). Inhibiting sulfate-reducing bacteria in biofilms by expressing the antimicrobial peptides indolicidin and bactenecin. Journal of Industrial Microbiology & Biotechnology, 22(3): 167–175

[7] Jia P, Dong L F, Tu Y, Diao Q Y. (2023). Bacillus subtilis and Macleaya cordata extract regulate the rumen microbiota associated with enteric methane emission in dairy cows. Microbiome, 11(1): 229

[8] Kim J K, Park K M. (2006). Effectiveness of Lactobacillus plantarum strain KJ-10311 to remove characteristic malodorous gases in piggery slurry. Asian-Australasian Journal of Animal Sciences, 19: 144–152

[9] Li X Y, Yang S F. (2007). Influence of loosely bound extracellular polymeric substances (EPS) on the flocculation, sedimentation and dewaterability of activated sludge. Water Research, 41(5): 1022–1030

[10] Liu S, Guo H, Wang Y, Hou J, Zhu T, Liu Y. (2023). Peracetic acid activated by ferrous ion mitigates sulfide and methane production in rising main sewers. Water Research, 245: 120584

[11] LiuY W, Ni B J, SharmaK R, YuanZ G (2015). Methane emission from sewers. Science of the Total Environment, 524–524: 40–51

[12] Lu L, Fang Y, Zhang W, Qi Q, Ju T, Chen J, Chen D Z. (2024). Biofilm regulation through biological autocrine signaling molecules and its deuterogenic benefits on gaseous dichloromethane degradation. Chemical Engineering Journal, 495: 153585

[13] Lu L, Ju T, Fang Y, Hu J, Sun Z, Cheng Z, Li Q, Chen J, Chen D Z. (2025). Efficient prediction of gaseous n-hexane removal in two-phase partitioning bioreactors with silicone oil based on the mechanism and kinetic models. Journal of Environmental Sciences, 154: 729–740

[14] Muller C, Guevarra K, Summers A, Pierce L, Shahbaz P, Zemke P E, Woodland K, Hollingsworth V, Nakhla G, Bell K. . (2022). A review of the practical application of micro-aeration and oxygenation for hydrogen sulfide management in anaerobic digesters. Process Safety and Environmental Protection, 165: 126–137

[15] Nghiem L D, Manassa P, Dawson M, Fitzgerald S K. (2014). Oxidation reduction potential as a parameter to regulate micro-oxygen injection into anaerobic digester for reducing hydrogen sulphide concentration in biogas. Bioresource Technology, 173: 443–447

[16] Olatunji K O, Ahmed N A, Ogunkunle O. (2021). Optimization of biogas yield from lignocellulosic materials with different pretreatment methods: a review. Biotechnology for Biofuels, 14(1): 159

[17] Qi Z, Sun N, Liu C. (2023). Glyoxylate cycle maintains the metabolic homeostasis of Pseudomonas aeruginosa in viable but nonculturable state induced by chlorine stress. Microbiological Research, 270: 127341

[18] Rathnayake D, Bal Krishna K C, Kastl G, Sathasivan A. (2021). The role of pH on sewer corrosion processes and control methods: a review. Science of the Total Environment, 782: 146616

[19] Shen Q, Redmile-Gordon M, Song J, Li J, Zhang K, Voroney P, Xu J, Brookes P C. (2021). Amendment with biodiesel co-product modifies genes for N cycling (nirK, nirS, nosZ) and greenhouse gas emissions (N₂O, CH₄, CO₂) from an acid soil. Biology and Fertility of Soils, 57(5): 629–642

[20] Sun J, Hu S, Sharma K R, Ni B J, Yuan Z. (2014). Stratified microbial structure and activity in sulfide- and methane-producing anaerobic sewer biofilms. Applied and Environmental Microbiology, 80(22): 7042–7052

[21] Valentino M J G, Kalaw S P, Galvez C T, Reyes R G. (2015). Mycota of distillery yeast sludge as source of single cell protein. Mycosphere: Journal of Fungal Biology, 6(3): 241–247

[22] Wang K, Huang D, Ying H, Luo H. (2014). Effects of acidification during storage on emissions of methane, ammonia, and hydrogen sulfide from digested pig slurry. Biosystems Engineering, 122: 23–30

[23] Wu B, Liu F, Fang W, Yang T, Chen G H, He Z, Wang S. (2021). Microbial sulfur metabolism and environmental implications. Science of the Total Environment, 778: 146085

[24] Wu H, Li A J, Zhang H W, Li S Q, Yang C Y, Lv H Y, Yao Y Q. (2024). Microbial mechanisms for higher hydrogen production in anaerobic digestion at constant temperature versus gradient heating. Microbiome, 12(1): 170

[25] Wu M, Wang T, Wu K, Kan L L. (2020). Microbiologically induced corrosion of concrete in sewer structures: a review of the mechanisms and phenomena. Construction & Building Materials, 239: 117813

[26] Yan R, Wu H, Yang X, Yang C, Lyu H, Zhang H, Li S, Liu T, Li R, Yao Y. (2023). Soil decreases N₂O emission and increases TN content during combined composting of wheat straw and cow manure by inhibiting denitrification. Chemical Engineering Journal, 477: 147306

[27] Yan Z Y, Liu X F, Yuan Y X, Liao Y Z, Li X D. (2013). Deodorization study of the swine manure with two yeast strains. Biotechnology and Bioprocess Engineering, 18(1): 135–143

[28] Yang X Y, Yan R X, Yang C Z, Zhang H W, Lyu H, Li S Q, Liu T R, Li R H, Yao Y Q, Li W T. . (2024). Soil accelerates the humification involved in co-composting of wheat straw and cattle manure by promoting humus formation. Chemical Engineering Journal, 479: 147583

[29] Zhang D, Pan X, Mostofa K M, Chen X, Mu G, Wu F, Liu J, Song W, Yang J, Liu Y, Fu Q. (2010). Complexation between Hg(II) and biofilm extracellular polymeric substances: an application of fluorescence spectroscopy. Journal of Hazardous Materials, 175(1−3): 359–365

[30] Zhou L Y, Xie Y Q, Wang X W, Wang Z F, Sa R, Li P B, Yang X P. (2024). Effect of microbial inoculation on nitrogen trans-formation, nitrogen functional genes, and bacterial community during cotton straw composting. Bioresource Technology, 403: 130859

[31] Zhuang R Y, Lou Y J, Qiu X T, Zhao Y Y, Qian D, Yan X J, He X D, Shen Q Q, Qian L Z. (2017). Identification of a yeast strain able to oxidize and remove sulfide high efficiently. Applied Microbiology and Biotechnology, 101(1): 391–400

[32] Zuo Z, Song Y, Ren D, Li H, Gao Y, Yuan Z, Huang X, Zheng M, Liu Y. (2020). Control sulfide and methane production in sewers based on free ammonia inactivation. Environment International, 143: 105928

[33] Zuo Z Q, Chang J, Lu Z S, Wang M R, Lin Y C, Zheng M, Zhu D Z, Yu T, Huang X, Liu Y C. (2019). Hydrogen sulfide generation and emission in urban sanitary sewer in China: What factor plays the critical role? Environmental Science. Water Research & Technology, 5(5): 839–848

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Higher Education Press 2025