Characteristics and influencing factors of biofilm formation by a pyridine-degrading bacterium Pseudomonas sp. ZX08

XIONG Fu-Zhong1 ZHAO Xiao-Xi1 WEN Dong-Hui1 LI Qi-Lin2

(1.College of Environmental Sciences and Engineering, Peking University, Beijing, China 100871)
(2.Department of Civil and Environmental Engineering, Rice University, Houston, TX 77005, USA)

【Abstract】[Background] The wastewater discharged by coal chemical industry contains a large number of refractory and highly toxic organic pollutants. It is an economically feasible strategy to treat the wastewater with bioaugmentation technology based on effective degrading bacteria. Promoting the biofilm formation of degrading bacteria is proved to be beneficial to the performance of the biofilm wastewater treatment system. [Objective] To investigate the biofilm formation process and characteristics of a pyridine-degrading bacterium Pseudomonas sp. ZX08, and to identify the influence of different environmental factors such as temperature, pH, Na+, K+, Ca2+, and Mg2+ on biofilm formation, and finally to provide reference for regulating biofilm formation in wastewater treatment systems. [Methods] A modified microtiter dish biofilm formation assay was used to determine the biofilm formation and the planktonic bacteria growth in the 12-well plate under different conditions; the structural characteristics of biofilm was observed using a confocal laser scanning microscope (CLSM). [Results] Pseudomonas sp. ZX08 showed good pyridine-degrading performance and biofilm-forming abilities. According to the CLSM analysis, the thickness of its biofilms formed at the surface reached 40–50 μm, and the proportion of live cells and protein/cell ratio were higher in the outer layer of the biofilms. A periodic variation was observed in the biofilm formation process in 72 h, and the biofilm biomass at the time points of 12 h and 48 h were relative peaks. The optimum temperature for ZX08 biofilm formation was 25°C, and the optimum pH range was 7.0–9.0. Higher concentrations of NaCl (> 0.6 mol/L) and KCl (> 0.4 mol/L) significantly inhibited the biofilm formation of ZX08. Within a certain range (0–16 mmol/L), the increase of Ca2+ concentration could promote the biofilm formation at the solid-liquid interface of the 12-well plate bottom. Adding 0–16 mmol/L Mg2+ also led to a slight increase in the biofilm formation of ZX08. [Conclusion] The pyridine-degrading bacterium Pseudomonas sp. ZX08 can form thick and stable biofilm, and it needs to comprehensively consider the influence of environmental factors on the biofilm formation of this strain in the future application.

【Keywords】 Pyridine; Effective degrading bacteria; Biofilm; Environmental factors;

【DOI】

【Funds】 National Natural Science Foundation of China (51378019, 51529801)

Download this article

    References

    [1] Herrero M, Stuckey DC. Bioaugmentation and its application in wastewater treatment: a review [J]. Chemosphere, 2015, 140: 119–128

    [2] Xie RS, Wu MM, Qu GF, et al. Treatment of coking wastewater by a novel electric assisted micro-electrolysis filter [J]. Journal of Environmental Sciences, 2018, 66: 165–172

    [3] Safwat SM. Performance of moving bed biofilm reactor using effective microorganisms [J]. Journal of Cleaner Production, 2018, 185: 723–731

    [4] He M, Zhang XJ, Qu FP, et al. Study on relativity between aerobic biodegradability and chemical structure of heterocyclic compounds [J]. China Environmental Science, 1997, 17 (3): 199–202 (in Chinese)

    [5] Kim MK, Singleton I, Yin CR, et al. Influence of phenol on the biodegradation of pyridine by freely suspended and immobilized Pseudomonas putida MK1 [J]. Letters in Applied Microbiology, 2006, 42 (5): 495–500

    [6] Bai YH, Sun QH, Zhao C, et al. Microbial degradation and metabolic pathway of pyridine by a Paracoccus sp. strain BW001 [J]. Biodegradation, 2008, 19 (6): 915–926

    [7] Sun QH, Bai YH, Zhao C, et al. Biodegradation of pyridine by Shinella zoogloeoides BC026 [J]. Environmental Science, 2008, 29 (10): 2938–2943 (in Chinese)

    [8] Jin TT, Ren JH, Zhang H, et al. Identification and characterization of a pyridine-degrading bacterium [J]. Ecology and Environmental Sciences, 2016, 25 (7): 1217–1224 (in Chinese)

    [9] Chen Q, Ni JR, Ma T, et al. Bioaugmentation treatment of municipal wastewater with heterotrophic-aerobic nitrogen removal bacteria in a pilot-scale SBR [J]. Bioresource Technology, 2015, 183: 25–32

    [10] Zhang J, Wen DH, Zhao C, et al. Bioaugmentation accelerates the shift of bacterial community structure against shock load: a case study of coking wastewater treatment by zeolite-sequencing batch reactor [J]. Applied Microbiology and Biotechnology, 2014, 98 (2): 863–873

    [11] Zhao B, Ran XC, Tian M, et al. Assessing the performance of a sequencing batch biofilm reactor bioaugmented with P. stutzeri strain XL-2 treating ammonium-rich wastewater [J]. Bioresource Technology, 2018, 270: 70–79

    [12] Bai YH, Sun QH, Sun RH, et al. Bioaugmentation and adsorption treatment of coking wastewater containing pyridine and quinoline using zeolite-biological aerated filters [J]. Environmental Science & Technology, 2011, 45 (5): 1940–1948

    [13] Yue WL, Chen M, Cheng ZQ, et al. Bioaugmentation of strain Methylobacterium sp. C1 towards p-nitrophenol removal with broad spectrum coaggregating bacteria in sequencing batch biofilm reactors [J]. Journal of Hazardous Materials, 2018, 344: 431–440

    [14] Yang GF, Feng LJ, Yang Q, et al. Startup pattern and performance enhancement of pilot-scale biofilm process for raw water pretreatment [J]. Bioresource Technology, 2014, 172: 22–31

    [15] Mao YJ, Quan X, Zhao HM, et al. Accelerated startup of moving bed biofilm process with novel electrophilic suspended biofilm carriers [J]. Chemical Engineering Journal, 2017, 315: 364–372

    [16] Flemming HC, Wingender J, Szewzyk U, et al. Biofilms: an emergent form of bacterial life [J]. Nature Reviews Microbiology, 2016, 14 (9): 563–575

    [17] Atkinson S, Williams P. Quorum sensing and social networking in the microbial world [J]. Journal of the Royal Society Interface, 2009, 6 (40): 959–978

    [18] Wu YC, Ding YZ, Cohen Y, et al. Elevated level of the second messenger c-di-GMP in Comamonas testosteroni enhances biofilm formation and biofilm-based biodegradation of 3-chloroaniline [J]. Applied Microbiology and Biotechnology, 2015, 99 (4): 1967–1976

    [19] Flemming HC, Neu TR, Wozniak DJ. The EPS matrix: the “House of Biofilm cells” [J]. Journal of Bacteriology, 2007, 189 (22): 7945–7947

    [20] Zhang N, Xiong FZ, Wen DH, et al. Effects of environmental factors on degrading bacterial biofilm formation [J]. Acta Scientiarum Naturalium Universitatis Pekinensis, 2016, 52 (2): 345–353 (in Chinese)

    [21] Xiong FZ, Zhao XX, Liao YH, et al. Effects of surface properties on biofilm formation and the related applications [J]. Microbiology China, 2018, 45 (1): 155–165 (in Chinese)

    [22] Zhao C, Wen DH, Zhang Y, et al. Experimental and mathematical methodology on the optimization of bacterial consortium for the simultaneous degradation of three nitrogen heterocyclic compounds [J]. Environmental Science & Technology, 2012, 46 (11): 6205–6213

    [23] Sezonov G, Joseleau-Petit D, D’Ari R. Escherichia coli physiology in Luria-Bertani broth [J]. Journal of Bacteriology, 2007, 189 (23): 8746–8749

    [24] Zhao C, Sun QH, Bai YH, et al. Physiological characteristics and application of pyridine-degrading bacterial strain Paracoccus sp. BW001 [J]. Environmental Pollution and Control, 2008, 30 (11): 17–22, 26 (in Chinese)

    [25] Dong XZ, Cai MY. Identification System Manual of Common Bacteria [M]. Beijing: Science Press, 2001 (in Chinese)

    [26] Rashid MH, Kornberg A. Inorganic polyphosphate is needed for swimming, swarming, and twitching motilities of Pseudomonas aeruginosa [J]. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97 (9): 4885–4890

    [27] O’Toole GA. Microtiter dish biofilm formation assay [J]. Journal of Visualized Experiments, 2011 (47): e2437

    [28] Guttenplan SB, Kearns DB. Regulation of flagellar motility during biofilm formation [J]. FEMS Microbiology Reviews, 2013, 37 (6): 849–871

    [29] Webb JS, Thompson LS, James S, et al. Cell death in Pseudomonas aeruginosa biofilm development [J]. Journal of Bacteriology, 2003, 185 (15): 4585–4592

    [30] Stoodley P, Sauer K, Davies DG, et al. Biofilms as complex differentiated communities [J]. Annual Review of Microbiology, 2002, 56: 187–209

    [31] Stanley NR, Lazazzera BA. Environmental signals and regulatory pathways that influence biofilm formation [J]. Molecular Microbiology, 2004, 52 (4): 917–924

    [32] Flemming HC, Wuertz S. Bacteria and archaea on earth and their abundance in biofilms [J]. Nature Reviews Microbiology, 2019, 17 (4): 247–260

    [33] Rinaudi L, Fujishige NA, Hirsch AM, et al. Effects of nutritional and environmental conditions on Sinorhizobium meliloti biofilm formation [J]. Research in Microbiology, 2006, 157 (9): 867–875

    [34] Zhou G, Li LJ, Shi QS, et al. Effects of nutritional and environmental conditions on planktonic growth and biofilm formation of Citrobacter werkmanii BF-6 [J]. Journal of Microbiology and Biotechnology, 2013, 23 (12): 1673–1682

    [35] Flemming HC, Wingender J. The biofilm matrix [J]. Nature Reviews Microbiology, 2010, 8 (9): 623–633

    [36] Qurashi AW, Sabri AN. Biofilm formation in moderately halophilic bacteria is influenced by varying salinity levels [J]. Journal of Basic Microbiology, 2012, 52 (5): 566–572

    [37] van der Waal SV, van der Sluis LWM, Özok AR, et al. The effects of hyperosmosis or high pH on a dual-species biofilm of Enterococcus faecalis and Pseudomonas aeruginosa: an in vitro study [J]. International Endodontic Journal, 2011, 44 (12): 1110–1117

    [38] Cogan NG, Keener JP. The role of the biofilm matrix in structural development [J]. Mathematical Medicine and Biology: A Journal of the IMA, 2004, 21 (2): 147–166

    [39] Janjaroen D, Ling FQ, Monroy G, et al. Roles of ionic strength and biofilm roughness on adhesion kinetics of Escherichia coli onto groundwater biofilm grown on PVC surfaces [J]. Water Research, 2013, 47 (7): 2531–2542

    [40] Geesey GG, Wigglesworth-Cooksey B, Cooksey KE. Influence of calcium and other cations on surface adhesion of bacteria and diatoms: a review [J]. Biofouling, 2000, 15 (1/3): 195–205

    [41] Shukla SK, Rao TS. Effect of calcium on Staphylococcus aureus biofilm architecture: a confocal laser scanning microscopic study [J]. Colloids and Surfaces B: Biointerfaces, 2013, 103: 448–454

    [42] Patrauchan MA, Sarkisova S, Sauer K, et al. Calcium influences cellular and extracellular product formation during biofilm-associated growth of a marine Pseudoalteromonas sp. [J]. Microbiology, 2005, 151 (9): 2885–2897

    [43] He XY, Wang JP, Abdoli L, et al. Mg2+/Ca2+ promotes the adhesion of marine bacteria and algae and enhances following biofilm formation in artificial seawater [J]. Colloids and Surfaces B: Biointerfaces, 2016, 146: 289–295

    [44] Gomes LC, Moreira JMR, Teodósio JS, et al. 96-well microtiter plates for biofouling simulation in biomedical settings [J]. Biofouling, 2014, 30 (5): 535–546

    [45] Ye YW, Ling N, Jiao R, et al. Effects of Ca2+ and Mg2+ on the biofilm formation of Cronobacter sakazakii strains from powdered infant formula [J]. Journal of Food Safety, 2015, 35 (3): 416–421

This Article

ISSN:0253-2654

CN: 11-1996/Q

Vol 47, No. 05, Pages 1342-1353

May 2020

Downloads:2

Share
Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results
  • 3 Discussion and conclusion
  • References