Enhancing biofilm formation in the hydrogen-based membrane biofilm reactor through bacterial Acyl-homoserine lactones

Abstract

● AHL-mediated quorum sensing is widely observed in H₂-denitrification systems.

● Inclusion of an external AHL source can enhance the induction of QS.

● C₁₄-HSL and C₄-HSL, especially C₁₄-HSL, can enhance biofilm formation.

● Tech to expedite autochthonous microbial biofilm formation has been proposed.

The slow growth rate of autotrophic bacteria and regulation of biofilm thickness are critical factors that limit the development of a hydrogen-based membrane biofilm reactor (H₂-MBfR). The acyl-homoserine lactone (AHL) mediated quorum sensing (QS) system is a crucial mechanism regulating biofilm behavior. However, the AHLs that promote biofilm formation in autotrophic denitrification systems and their underlying mechanisms, remain unclear. This study explored the impact of AHL-mediated QS signaling molecules on biofilm development in H₂-MBfR. This study revealed that C₁₄-HSL and C₄-HSL are potential signaling molecules that enhanced biofilm formation in long-term stable operating H₂-MBfR. Subsequent short-term experiments with C₁₄-HSL and C₄-HSL confirmed their ability to increase bacterial adhesion to carrier surfaces by promoting the production of extracellular polymeric substances (EPS). Functional gene annotation indicated that exogenous C₁₄-HSL and C₄-HSL increased the abundance of signal transduction (increased by 0.250%–0.375%), strengthening the inter bacterial QS response while enhancing cell motility (increased by 0.24% and 0.21%, respectively) and biological adhesion (increased by 0.044% and 0.020%, respectively), thereby accelerating the initial bacterial attachment to hollow fiber membranes and facilitating biofilm development. These findings contribute to the understanding of microbial community interactions in H₂-MBfRs and provide novel approaches for biofilm management in wastewater treatment systems.

Graphical abstract

Keywords

Hydrogen-based membrane biofilm reactor (H ₂-MBfR) / Acyl-homoserine lactones (AHLs) / Quorum sensing (QS) / Biofilm enhancement

References

[1] Bolger A M, Lohse M, Usadel B. (2014). Trimmomatic: a flexible trimmer for illumina sequence data. Bioinformatics, 30(15): 2114–2120

[2] Cataldi T R, Bianco G, Palazzo L, Quaranta V. (2007). Occurrence of N-acyl-L-homoserine lactones in extracts of some Gram-negative bacteria evaluated by gas chromatography–mass spectrometry. Analytical Biochemistry, 361(2): 226–235

[3] Elsayed A, Hurdle M, Kim Y. (2021). Comprehensive model applications for better understanding of pilot-scale membrane-aerated biofilm reactor performance. Journal of Water Process Engineering, 40: 101894

[4] Feng Q, Luo L, Chen X, Zhang K, Fang F, Xue Z, Li C, Cao J, Luo J. (2021). Facilitating biofilm formation of Pseudomonas aeruginosa via exogenous N-Acy-L-homoserine lactones stimulation: regulation on the bacterial motility, adhesive ability and metabolic activity. Bioresource Technology, 341: 125727

[5] Feng Z, Lu X, Chen C, Huo Y, Zhou D. (2022). Transboundary intercellular communications between Penicillium and bacterial communities during sludge bulking: inspirations on quenching fungal dominance. Water Research, 221: 118829

[6] Feng Z, Sun Y, Li T, Meng F, Wu G. (2019). Operational pattern affects nitritation, microbial community and quorum sensing in nitrifying wastewater treatment systems. Science of the Total Environment, 677: 456–465

[7] Gao M, Peng Y, Shen Y, Tan F. (2023). Study of the biofilm mechanism of C₄-HSL and C₆-HSL in the degradation of quinoline. Journal of Biotechnology, 376: 53–63

[8] González A, Bellenberg S, Mamani S, Ruiz L, Echeverría A, Soulère L, Doutheau A, Demergasso C, Sand W, Queneau Y. . (2013). AHL signaling molecules with a large acyl chain enhance biofilm formation on sulfur and metal sulfides by the bioleaching bacterium Acidithiobacillus ferrooxidans. Applied Microbiology and Biotechnology, 97(8): 3729–3737

[9] Hou F, Zhang T, Peng Y, Cao X, Pang H, Shao Y, Lu X, Yuan J, Chen X, Zhang J. (2022). Partial anammox achieved in full scale biofilm process for typical domestic wastewater treatment. Frontiers of Environmental Science & Engineering, 16(3): 33

[10] Hu H, He J, Liu J, Yu H, Zhang J. (2016). Biofilm activity and sludge characteristics affected by exogenous N-acyl homoserine lactones in biofilm reactors. Bioresource Technology, 211: 339–347

[11] Huang J, Shi Y, Zeng G, Gu Y, Chen G, Shi L, Hu Y, Tang B, Zhou J. (2016). Acyl-homoserine lactone-based quorum sensing and quorum quenching hold promise to determine the performance of biological wastewater treatments: an overview. Chemosphere, 157: 137–151

[12] Huang W, Gong B, He L, Wang Y, Zhou J. (2020). Intensified nutrients removal in a modified sequencing batch reactor at low temperature: metagenomic approach reveals the microbial community structure and mechanisms. Chemosphere, 244: 125513

[13] Hwang B K, Lee W N, Park P K, Lee C H, Chang I S. (2007). Effect of membrane fouling reducer on cake structure and membrane permeability in membrane bioreactor. Journal of Membrane Science, 288(1−2): 149–156

[14] Jiang M, Zhang Y, Zheng J, Li H, Ma J, Zhang X, Wei Q, Wang X, Zhang X, Wang Z. (2022). Mechanistic insights into CO₂ pressure regulating microbial competition in a hydrogen-based membrane biofilm reactor for denitrification. Chemosphere, 303: 134875

[15] Jiang M, Zheng J, Perez-Calleja P, Picioreanu C, Lin H, Zhang X, Zhang Y, Li H, Nerenberg R. (2020). New insight into CO₂-mediated denitrification process in H₂-based membrane biofilm reactor: an experimental and modeling study. Water Research, 184: 116177

[16] Lai C Y, Zhong L, Zhang Y, Chen J X, Wen L L, Shi L D, Sun Y P, Ma F, Rittmann B E, Zhou C. . (2016). Bioreduction of chromate in a methane-based membrane biofilm reactor. Environmental Science & Technology, 50(11): 5832–5839

[17] Lee K, Yu H, Zhang X, Choo K H. (2018). Quorum sensing and quenching in membrane bioreactors: opportunities and challenges for biofouling control. Bioresource Technology, 270: 656–668

[18] Li H, Han Y, Zhang Y, Mi X, Wang D, Xu Y, Dong K. (2024). Optimization of nitrogen removal and microbial mechanism of a hydrogen-based membrane biofilm reactor. Environmental Technology, 46: 1–17

[19] Li J, Ma J, Sun L, Liu X, Liao H, He D. (2022). Mechanistic insight into the biofilm formation and process performance of a passive aeration ditch (PAD) for decentralized wastewater treatment. Frontiers of Environmental Science & Engineering, 16(7): 86

[20] Li T, Guo F, Lin Y, Li Y, Wu G. (2019). Metagenomic analysis of quorum sensing systems in activated sludge and membrane biofilm of a full-scale membrane bioreactor. Journal of Water Process Engineering, 32: 100952

[21] Li T, Yang B, Li X, Li J, Zhao G, Kan J. (2018). Quorum sensing system and influence on food spoilage in Pseudomonas fluorescens from turbot. Journal of Food Science and Technology, 55(8): 3016–3025

[22] Li Y, Xia H, Bai F, Song X, Zhuang L, Xu H, Zhang X, Zhang X, Qiao M. (2020). PA5001 gene involves in swimming motility and biofilm formation in Pseudomonas aeruginosa. Microbial Pathogenesis, 144: 103982

[23] Liu Q, Wang J, He R, Hu H, Wu B, Ren H. (2020). Bacterial assembly during the initial adhesion phase in wastewater treatment biofilms. Water Research, 184: 116147

[24] Lu X, Yan G, Fu L, Cui B, Wang J, Zhou D. (2023). A review of filamentous sludge bulking controls from conventional methods to emerging quorum quenching strategies. Water Research, 236: 119922

[25] Lynch M J, Swift S, Kirke D F, Keevil C W, Dodd C E, Williams P. (2002). The regulation of biofilm development by quorum sensing in Aeromonas hydrophila. Environmental Microbiology, 4(1): 18–28

[26] Mukherjee S, Bassler B L. (2019). Bacterial quorum sensing in complex and dynamically changing environments. Nature Reviews. Microbiology, 17(6): 371–382

[27] Ni S Q, Sun N, Yang H, Zhang J, Ngo H H. (2015). Distribution of extracellular polymeric substances in anammox granules and their important roles during anammox granulation. Biochemical Engineering Journal, 101: 126–133

[28] Oh H S, Lee C H. (2018). Origin and evolution of quorum quenching technology for biofouling control in MBRs for wastewater treatment. Journal of Membrane Science, 554: 331–345

[29] Pang Y, Wang S, Tao J, Wang J, Xue Z, Wang R. (2022). Mechanism of berberine hydrochloride interfering with biofilm formation of Hafnia alvei. Archives of Microbiology, 204(2): 126–134

[30] Rittmann B E. (2018). Biofilms, active substrata, and me. Water Research, 132: 135–145

[31] Sun Y, Guan Y, Zeng D, He K, Wu G. (2018). Metagenomics-based interpretation of AHLs-mediated quorum sensing in anammox biofilm reactors for low-strength wastewater treatment. Chemical Engineering Journal, 344: 42–52

[32] Sun Z, Xi J, Yang C, Cong W. (2022). Quorum sensing regulation methods and their effects on biofilm in biological waste treatment systems: a review. Frontiers of Environmental Science & Engineering, 16(7): 87

[33] Sun Z, Yang B, Yeung M, Xi J. (2023). Effects of exogenous acylated homoserine lactones on biofilms in biofilters for gaseous toluene treatment. Frontiers of Environmental Science & Engineering, 17(2): 17

[34] TanC H, Yeo Y P, HafizM, NgN K J, Subramoni S, TajS, TayM, ChaoX, KjellebergS, Rice S A (2021). Functional metagenomic analysis of quorum sensing signaling in a nitrifying community. npj Biofilms and Microbiomes, 7(1): 79

[35] Tomlin K L, Malott R J, Ramage G, Storey D G, Sokol P A, Ceri H. (2005). Quorum-sensing mutations affect attachment and stability of Burkholderia cenocepacia biofilms. Applied and Environmental Microbiology, 71(9): 5208–5218

[36] Wang J, Jiang Z, Wang W, Wang H, Zhang Y, Wang Y. (2021a). The connection between aeration regimes and EPS composition in nitritation biofilm. Chemosphere, 265: 129–141

[37] Wang J, Liu Q, Dong D, Hu H, Wu B, Ren H. (2021b). AHLs-mediated quorum sensing threshold and its response towards initial adhesion of wastewater biofilms. Water Research, 194: 116925

[38] Wang J, Ren H, Li X, Li J, Ding L, Geng J, Xu K, Huang H, Hu H. (2018). In situ monitoring of wastewater biofilm formation process via ultrasonic time domain reflectometry (UTDR). Chemical Engineering Journal, 334: 2134–2141

[39] Wang X, Jiang C, Wang D, Yang Y, Fan L, Xu S, Zhuang X. (2023). Quorum sensing responses of activated sludge to free nitrous acid: Zoogloea deformation, AHL redistribution, and microbiota acclimatization. Water Research, 238: 119993

[40] Wu B, Chai X, Zhao Y. (2016). Enhanced dewatering of waste-activated sludge by composite hydrolysis enzymes. Bioprocess and Biosystems Engineering, 39(4): 627–639

[41] Wu B, Wang H, Dai X, Chai X. (2021). Influential mechanism of water occurrence states of waste-activated sludge: specifically focusing on the roles of EPS micro-spatial distribution and cation-dominated interfacial properties. Water Research, 202: 117461

[42] Wu Y, Li Y, Ontiveros-Valencia A, Ordaz-Díaz L, Liu J, Zhou C, Rittmann B E. (2017). Enhancing denitrification using a novel in situ membrane biofilm reactor (isMBfR). Water Research, 119: 234–241

[43] Yu H, Xu G, Qu F, Li G, Liang H. (2016). Effect of solid retention time on membrane fouling in membrane bioreactor: from the perspective of quorum sensing and quorum quenching. Applied Microbiology and Biotechnology, 100(18): 7887–7897

[44] Yue Z, Li P, Bin L, Huang S, Fu F, Yang Z, Qiu B, Tang B. (2020). N-Acyl-homoserine lactone-mediated quorum sensing of aerobic granular sludge system in a continuous-flow membrane bioreactor. Biochemical Engineering Journal, 164: 107801

[45] Zhang Y, Chen J X, Wen L L, Tang Y, Zhao H P. (2016). Effects of salinity on simultaneous reduction of perchlorate and nitrate in a methane-based membrane biofilm reactor. Environmental Science and Pollution Research International, 23(23): 24248–24255

[46] Zhang Y, Jiang M, Ma J, Wang Y, Zhang X, Wei Q, Wang X, Zhang X, Zheng J. (2023a). Development of a novel periodic venting-controlled membrane biofilm reactor for hydrogenotrophic denitrification: process performance and microbial mechanism. Chemical Engineering Journal, 463: 142529

[47] Zhang Y, Yu H, Xie Y, Guo Y, Cheng Y, Yao W. (2023b). Inhibitory effects of hexanal on acylated homoserine lactones (AHLs) production to disrupt biofilm formation and enzymes activity in Erwinia carotovora and Pseudomonas fluorescens. Journal of Food Science and Technology, 60(1): 372–381

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