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Persistence of V. cholerae O139 Biofilm Against Physical, Chemical and Antibiotic Lethal Challenges

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Since its emergence, cholera caused by the bacterium Vibrio cholerae remains as a significant threat to human health. The continued persistence of this pathogen against many unfavourable conditions made challenging to eradicate cholera, especially in the developing countries. In the hostile conditions, the bacterium is known to form a self-enclosed polymeric structure called ‘biofilm’ which serves as a major factor responsible for its persistence and transmission. Hence, the authors aimed to understand the effectiveness of relevant physical, chemical and antibiotic treatments against the biofilm of this bacterium. For the study, the biofilm of V. cholerae O139 wild type and its isogenic Tn5-mutants that differ in their biofilm phenotype (biofilm proficient and biofilm deficient) were exposed to different levels of pH, salinity, temperature, UV radiation, H2O2, chlorination, and antibiotics. It was observed that biofilm culture of both wild type and the biofilm-proficient mutant exhibited detectable survival rate up to pH 2.0, salinity 3.5 M, temperature 50 °C, H2O2 80 mM, NaOCl 12.5 mg/L and at several folds increased antibiotic concentration (Ciprofloxacin 2 mg/L, and Doxycycline 256 mg/L) as compared to the biofilm defective mutant and the planktonic cultures of respective strains. As the biofilm of the cholera pathogen or the cholera biofilm resists several lethal challenges, it is essential to target the biofilm and its residents for the complete destruction of the infection source and thereby to prevent transmission of cholera.

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References

  1. Lakshminarayanan S, Jayalakshmy R (2015) Diarrheal diseases among children in India: current scenario and future perspectives. J Nat Sci Bio Med 6:24–28. https://doi.org/10.4103/0976-9668.149073

    Article  Google Scholar 

  2. World Health Organization (2017) Diarrhoeal diseases. http://www.who.int/mediacentre/factsheets/fs330/en/. Accessed 2 May 2017

  3. Chakraborty S, Nair GB, Shinoda S (1997) Pathogenic vibrios in the natural aquatic environment. Rev Env Health 12:63–80

    Article  CAS  Google Scholar 

  4. Faruque SM, Albert MJ, Mekalanos JJ (1998) Epidemiology, genetics, and ecology of toxigenic Vibrio cholerae. Microbiol Mol Biol Rev 62:1301–1314

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Gil AI, Louis VR, Rivera ING, Lipp E, Huq A, Lanata CF, Colwell RR (2004) Occurrence and distribution of Vibrio cholerae in the coastal environment of Peru. Environ Microbiol 6:699–706. https://doi.org/10.1111/j.1462-2920.2004.00601.x

    Article  PubMed  Google Scholar 

  6. Tamplin ML, Gauzens AL, Huq A, Sack DA, Colwell RR (1990) Attachment of Vibrio cholerae serogroup O1 to zooplankton and phytoplankton of Bangladesh waters. Appl Environ Microbiol 56:1977–1980

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Watnick PI, Kolter R (1999) Steps in the development of a Vibrio cholerae El Tor biofilm. Mol Microbiol 34:586–595. https://doi.org/10.1046/j.1365-2958.1999.01624.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Yildiz FH, Schoolnik GK (1999) Vibrio cholerae O1 El Tor: identification of a gene cluster required for the rugose colony type, exopolysaccharide production, chlorine resistance, and biofilm formation. Proc Natl Acad Sci USA 96:4028–4033. https://doi.org/10.1073/pnas.96.7.4028

    Article  CAS  PubMed  Google Scholar 

  9. Castro-Rosas J, Escartín EF (2005) Increased tolerance of Vibrio cholerae O1 to temperature, pH, or drying associated with colonization of shrimp carapaces. Int J Food Microbiol 102:195–201. https://doi.org/10.1016/j.ijfoodmicro.2004.12.015

    Article  CAS  PubMed  Google Scholar 

  10. Tamayo R, Patimalla B, Camilli A (2010) Growth in a biofilm induces a hyperinfectious phenotype in Vibrio cholerae. Infect Immun 78:3560–3569. https://doi.org/10.1128/IAI.00048-10

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Akanda AS, Jutla AS, Alam M et al (2011) Hydroclimatic influences on seasonal and spatial cholera transmission cycles: implications for public health intervention in the Bengal Delta. Water Resour Res 47:W00H07. https://doi.org/10.1029/2010WR009914

    Article  Google Scholar 

  12. Huq A, Sack RB, Nizam A, Longini IM, Nair GB, Ali A, Morris JG, Khan MNH, Siddique AK, Yunus M, Albert MJ, Sack DA, Colwell RR (2005) Critical factors influencing the occurrence of Vibrio cholerae in the environment of Bangladesh. Appl Environ Microbiol 71:4645–4654

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Gupta P, Mankere B, Chekkoora Keloth S, Tuteja U, Chelvam KT (2018) Generation and in vivo characterization of Tn5-induced biofilm mutants of Vibrio cholerae O139. Curr Microbiol. https://doi.org/10.1007/s00284-018-1527-8

    Article  PubMed  Google Scholar 

  14. Singh A, Barnard TG (2016) Surviving the acid barrier: responses of pathogenic Vibrio cholerae to simulated gastric fluid. Appl Microbiol Biotechnol 100:815–824. https://doi.org/10.1007/s00253-015-7067-2

    Article  CAS  PubMed  Google Scholar 

  15. Clinical and Laboratory Standards Institute (2017) Performance standards for antimicrobial susceptibility testing: twenty-seven informational supplement M100–S27. CLSI, Wayne, PA, USA, 2017

    Google Scholar 

  16. Faruque SM, Biswas K, Udden SM, Ahmad QS, Sack DA, Nair GB, Mekalanos JJ (2006) Transmissibility of cholera: invivo-formed biofilms and their relationship to infectivity and persistence in the environment. Proc Natl Acad Sci USA 103:6350–6355. https://doi.org/10.1073/pnas.0601277103

    Article  CAS  PubMed  Google Scholar 

  17. Nelson EJ, Chowdhury A, Harris JB et al (2007) Complexity of rice water stool from patients with Vibrio cholerae plays a role in the transmission of infectious diarrhoea. Proc Natl Acad Sci USA 104:19091–19096. https://doi.org/10.1073/pnas.0706352104

    Article  PubMed  Google Scholar 

  18. Teschler JK, Zamorano-Sanchez D, Utada AS et al (2015) Living in the matrix: assembly and control of Vibrio cholerae biofilms. Nat Rev Microbiol 13:255–268. https://doi.org/10.1038/nrmicro3433

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Wang Y, Leung PC, Qian P, Gu JD (2004) Effects of UV, H2O2 and Fe3+ on the growth of four environmental isolates of Aeromonas and Vibrio species from a Mangrove environment. Microbes Environ 19:163–171

    Article  Google Scholar 

  20. Russell AD (2003) Lethal effects of heat on bacterial physiology and structure. Sci Prog 86:115–137

    Article  CAS  PubMed  Google Scholar 

  21. Wilson BR, Roessler PF, VanDellen E, Abbaszadegan M, Gerba CP (1992) Coliphage MS-2 as a UV water disinfection efficacy test surrogate for bacterial and viral pathogens. In: Proceedings, water quality technology conference, November 15–19 Toronto, Canada, pp 219–235

  22. Jacobs JL, Sundin GW (2001) Effect of solar UV-B radiation on a phyllosphere bacterial community. Appl Environ Microbiol 67:5488–5496. https://doi.org/10.1128/AEM.67.12.5488-5496.2001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Sinha RP, Häder DP (2002) UV-induced DNA damage and repair: a review. Photochem Photobiol Sci 1:225–236

    Article  CAS  PubMed  Google Scholar 

  24. Brioukhanov AL, Netrusov AI (2004) Catalase and superoxide dismutase: distribution, properties, and physiological role in cells of strict anaerobes. Biochemistry (Mosc) 69:949–962

    Article  CAS  Google Scholar 

  25. van Vliet AH, Ketley JM, Park SF, Penn CW (2002) The role of iron in Campylobactergene regulation, metabolism and oxidative stress defense. FEMS Microbiol Rev 26:173–186

    Article  PubMed  Google Scholar 

  26. Barnes AC, Bowden TJ, Horne MT, Ellis AE (1999) Peroxide-inducible catalase in Aeromonas salmonicida subsp. salmonicida protects against exogenous hydrogen peroxide and killing by activated rainbow trout, Oncorhynchus mykiss L., macrophages. Microb Pathogen 26:149–158

    Article  CAS  Google Scholar 

  27. Yildiz FH, Schoolnik GK (1998) Role of rpoS in stress survival and virulence of Vibrio cholerae. J Bacteriol 180:773–784

    CAS  PubMed  PubMed Central  Google Scholar 

  28. International Commission of Microbiological Specification for Food (1974) Microorganisms in food -2. University of Toronto Press, Toronto

    Google Scholar 

  29. National Research Council (1980) Drinking water and heath, vol 2. DC National academies press (US), Washington. https://doi.org/10.17226/1904

    Book  Google Scholar 

  30. World Health Organization & International Programme on Chemical Safety (1996) Guidelines for drinkingwater quality, 2nd ed. Health criteria and other supporting information, vol 2. World Health Organization, Geneva. http://www.who.int/iris/handle/10665/38551

  31. Silva AJ, Benitez JA (2016) Vibrio cholerae biofilms and cholera pathogenesis. PLoS Negl Trop Dis 10:e0004330. https://doi.org/10.1371/journal.pntd.0004330

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Sengupta C, Mukherjee O, Chowdhury R (2016) Adherence to intestinal cells promotes biofilm formation in Vibrio cholerae. J Infect Dis 214:1571–1578

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors are thankful to the Director, Defence Research and Development Establishment (DRDE) Gwalior, the Government of India (DRDE/MB/011/2018), for providing all the facilities and financial support to carry out this work. They are extremely grateful to Prof. Eric V. Stabb, Department of Microbiology, University of Georgia, Athens Georgia, for the kind gift of pEVS 168, pEVS104 vectors, and E. coli DH5 α/λpir host, which were used in the construction biofilm, altered Tn5 mutants of V. cholerae O139. This study was supported by Institutional fund provided by Defence R&D Establishment, Gwalior, Ministry of Defence, the Government of India.

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Correspondence to Kulanthaivel Thava Chelvam.

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Significance statement The V. cholerae O139 biofilm was found to be more resistant to high temperature, UV radiation, low pH, salinity, oxidative treatment, chlorination, and antibiotic treatments. It is essential to address the biofilm of V. cholerae for effective control.

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Gupta, P., Mankere, B., Keloth, S.C. et al. Persistence of V. cholerae O139 Biofilm Against Physical, Chemical and Antibiotic Lethal Challenges. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 1117–1124 (2019). https://doi.org/10.1007/s40011-018-1032-7

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  • DOI: https://doi.org/10.1007/s40011-018-1032-7

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