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Chlorine disinfection promotes the exchange of antibiotic resistance genes across bacterial genera by natural transformation

Min Jin, Lu Liu, Da-ning Wang and 11 more

The ISME Journal · 2020 · CC BY

doi:10.1038/s41396-020-0656-9

Abstract

Chlorine disinfection to drinking water plays an important role in preventing and controlling waterborne disease outbreaks globally. Nevertheless, little is known about why it enriches the antibiotic resistance genes (ARGs) in bacteria after chlorination. Here, ARGs released from killed antibiotic-resistant bacteria (ARB), and culturable chlorine-injured bacteria produced in the chlorination process as the recipient, were investigated to determine their contribution to the horizontal transfer of ARGs during disinfection treatment. We discovered Escherichia coli, Salmonella aberdeen, Pseudomonas aeruginosa and Enterococcus faecalis showed diverse resistance to sodium hypochlorite, and transferable RP4 could be released from killed sensitive donor consistently. Meanwhile, the survival of chlorine-tolerant injured bacteria with enhanced cell membrane permeabilisation and a strong oxidative stress-response demonstrated that a physiologically competent cell could be transferred by RP4 with an improved transformation frequency of up to 550 times compared with the corresponding untreated bacteria. Furthermore, the water quality factors involving chemical oxygen demand (CODMn), ammonium nitrogen and metal ions (Ca2+ and K+) could significantly promote above transformation frequency of released RP4 into injured E. faecalis. Our findings demonstrated that the chlorination process promoted the horizontal transfer of plasmids by natural transformation, which resulted in the exchange of ARGs across bacterial genera and the emergence of new ARB, as well as the transfer of chlorine-injured opportunistic pathogen from non-ARB to ARB. Considering that the transfer elements were quite resistant to degradation through disinfection, this situation poses a potential risk to public health.

References 45

  1. 1.

    Pruden A, Pei R, Storteboom H, Carlson KH. Antibiotic resistance genes as emerging contaminants: studies in northern Colorado. Environ Sci Technol. 2006;40:7445-50.

    Not in our library10.1021/es060413l

  2. 2.

    O’Neill J. Antimicrobial resistance: tackling a crisis for the health and wealth of nations. 2015.

    Not in our library

  3. 3.

    Watkins RR, Bonomo RA. Overview: global and local impact of antibiotic resistance. Infect Dis Clin N Am. 2016;30:313-22.

    Not in our library10.1016/j.idc.2016.02.001

  4. 4.

    Zhu YG, Gillings M, Simonet P, Stekel D, Banwart S, Penuelas J. Microbial mass movements. Science. 2017;357:1099-100.

    Not in our library10.1126/science.aao3007

  5. 5.

    Fricker CR, Eldred BJ. The effect of sodium azide concentration on the recovery of Enterococci from water. J Water Health. 2014;12:264-8.

    Not in our library10.2166/wh.2013.422

  6. 6.

    Wang X, Devlieghere F, Geeraerd A, Uyttendaele M. Thermal inactivation and sublethal injury kinetics of Salmonella enterica and Listeria monocytogenes in broth versus agar surface. Int J Food Microbiol. 2017;243:70-7.

    Not in our library10.1016/j.ijfoodmicro.2016.12.008

  7. 7.

    Ano T, Shoda M. Ultra-rapid transformation of Escherichia coli by an alkali cation. Biosci Biotechnol Biochem. 1992;56:1505.

    Not in our library10.1271/bbb.56.1505

  8. 8.

    Baquero F, Martinez JL, Canton R. Antibiotics and antibiotic resistance in water environments. Curr Opin Biotechnol. 2008;19:260-5.

    Not in our library10.1016/j.copbio.2008.05.006

  9. 9.

    Marti E, Variatza E, Balcazar JL. The role of aquatic ecosystems as reservoirs of antibiotic resistance. Trends Microbiol. 2014;22:36-41.

    Not in our library10.1016/j.tim.2013.11.001

  10. 10.

    Karkman A, Do TT, Walsh F, Virta MPJ. Antibiotic-resistance genes in waste water. Trends Microbiol. 2018;26:220-8.

    Not in our library10.1016/j.tim.2017.09.005

  11. 11.

    Czekalski N, Gascon Diez E, Burgmann H. Wastewater as a point source of antibiotic-resistance genes in the sediment of a freshwater lake. ISME J. 2014;8:1381-90.

    Not in our library10.1038/ismej.2014.8

  12. 12.

    Yan L, Liu D, Wang XH, Wang Y, Zhang B, Wang M, et al. Bacterial plasmid-mediated quinolone resistance genes in aquatic environments in China. Sci Rep. 2017;7:40610.

    Not in our library10.1038/srep40610

  13. 13.

    Li J, Liu L, Yang D, Liu WL, Shen ZQ, Qu HM, et al. Culture-dependent enumeration methods failed to simultaneously detect disinfectant-injured and genetically modified Escherichia coli in drinking water. Environ Sci Process Impacts. 2017;19:720-6.

    Not in our library10.1039/c6em00625f

  14. 14.

    Hasegawa H, Suzuki E, Maeda S. Horizontal plasmid transfer by transformation in Escherichia coli: Environmental factors and possible mechanisms. Front Microbiol. 2018;9:2365.

    Not in our library10.3389/fmicb.2018.02365

  15. 15.

    Nishioka M, Mashayekhan S, Onishi K, Taya M. The biofilm environment offers a possible condition for inducing the competency of DNA recipient cells through nutritional starvation. Biocontrol Sci. 2007;12:115-8.

    Not in our library10.4265/bio.12.115

  16. 16.

    Khan S, Beattie TK, Knapp CW. Relationship between antibiotic- and disinfectant-resistance profiles in bacteria harvested from tap water. Chemosphere. 2016;152:132-41.

    Not in our library10.1016/j.chemosphere.2016.02.086

  17. 17.

    McCurry DL, Krasner SW, von Gunten U, Mitch WA. Determinants of disinfectant pretreatment efficacy for nitrosamine control in chloraminated drinking water. Water Res. 2015;84:161-70.

    Not in our library10.1016/j.watres.2015.07.024

  18. 18.

    Nwachcuku N, Gerba CP. Emerging waterborne pathogens: can we kill them all? Curr Opin Biotechnol. 2004;15:175-80.

    Not in our library10.1016/j.copbio.2004.04.010

  19. 19.

    Liu SS, Qu HM, Yang D, Hu H, Liu WL, Qiu ZG, et al. Chlorine disinfection increases both intracellular and extracellular antibiotic resistance genes in a full-scale wastewater treatment plant. Water Res. 2018;136:131-6.

    Not in our library10.1016/j.watres.2018.02.036

  20. 20.

    Shi P, Jia S, Zhang XX, Zhang T, Cheng S, Li A. Metagenomic insights into chlorination effects on microbial antibiotic resistance in drinking water. Water Res. 2013;47:111-20.

    Not in our library10.1016/j.watres.2012.09.046

  21. 21.

    Zheng DW, Chen JL, Zhu JY, Rong L, Li B, Lei Q, et al. Highly integrated nano-platform for breaking the barrier between chemotherapy and immunotherapy. Nano Lett. 2016;16:4341-7.

    Not in our library10.1021/acs.nanolett.6b01432

  22. 22.

    Xi C, Zhang Y, Marrs CF, Ye W, Simon C, Foxman B, et al. Prevalence of antibiotic resistance in drinking water treatment and distribution systems. Appl Environ Microbiol. 2009;75:5714-8.

    Not in our library10.1128/aem.00382-09

  23. 23.

    Baur B, Hanselmann K, Schlimme W, Jenni B. Genetic transformation in freshwater: Escherichia coli is able to develop natural competence. Appl Environ Microbiol. 1996;62:3673-8.

    Not in our library10.1128/aem.62.10.3673-3678.1996

  24. 24.

    Dodd MC. Potential impacts of disinfection processes on elimination and deactivation of antibiotic resistance genes during water and wastewater treatment. J Environ Monit. 2012;14:1754-71.

    Not in our library10.1039/c2em00006g

  25. 25.

    Hao H, Shi DY, Yang D, Yang ZW, Qiu ZG, Liu WL, et al. Profiling of intracellular and extracellular antibiotic resistance genes in tap water. J Hazard Mater. 2019;365:340-5.

    Not in our library10.1016/j.jhazmat.2018.11.004

  26. 26.

    Bissonnette G, Jezeski J, McFeters G, Stuart D. Influence of environmental stress on enumeration of indicator bacteria from natural waters. Appl Microbiol. 1975;29:186-94.

    Not in our library10.1128/am.29.2.186-194.1975

  27. 27.

    McFeters GA, LeChevallier MW, Singh A, Kippin JS. Health significance and occurrence of injured bacteria in drinking water. Water Sci Technol. 1986;18:227-31.

    Not in our library

  28. 28.

    Hou AM, Yang D, Miao J, Shi DY, Yin J, Yang ZW, et al. Chlorine injury enhances antibiotic resistance in Pseudomonas aeruginosa through over expression of drug efflux pumps. Water Res. 2019;156:366-71.

    Not in our library10.1016/j.watres.2019.03.035

  29. 29.

    Ch’ng JH, Chong KKL, Lam LN, Wong JJ, Kline KA. Biofilm-associated infection by Enterococci. Nat Rev Microbiol. 2019;17:82-94.

    Not in our library10.1038/s41579-018-0107-z

  30. 30.

    Kao PHN, Kline KA. Dr. Jekyll and Mr. Hide: how Enterococcus faecalis subverts the host immune response to cause infection. J Mol Biol. 2019;431:2932-45.

    Not in our library10.1016/j.jmb.2019.05.030

  31. 31.

    American Public Health Association (APHA). Standard methods for the examination of water and wastewater. 1998.

    Not in our library

  32. 32.

    Froger A, Hall JE. Transformation of plasmid DNA into E. coli using the heat shock method. J Vis Exp. 2007:253.

    Not in our library10.3791/253

  33. 33.

    Martinez JL. Environmental pollution by antibiotics and by antibiotic resistance determinants. Environ Pollut. 2009;157:2893-902.

    Not in our library10.1016/j.envpol.2009.05.051

  34. 34.

    MacFadden DR, McGough SF, Fisman D, Santillana M, Brownstein JS. Antibiotic resistance increases with local temperature. Nat Clim Chang. 2018;8:510-4.

    Not in our library10.1038/s41558-018-0161-6

  35. 35.

    Murphy EA, Post GB, Buckley BT, Lippincott RL, Robson MG. Future challenges to protecting public health from drinking-water contaminants. Annu Rev Public Health. 2012;33:209-24.

    Not in our library10.1146/annurev-publhealth-031811-124506

  36. 36.

    Stange C, Sidhu JPS, Toze S, Tiehm A. Comparative removal of antibiotic resistance genes during chlorination, ozonation, and UV treatment. Int J Hyg Environ Health. 2019;222:541-8.

    Not in our library10.1016/j.ijheh.2019.02.002

  37. 37.

    He H, Zhou P, Shimabuku KK, Fang X, Li S, Lee Y, et al. Degradation and deactivation of bacterial antibiotic resistance genes during exposure to free chlorine, monochloramine, chlorine dioxide, ozone, ultraviolet light, and hydroxyl radical. Environ Sci Technol. 2019;53:2013-26.

    Not in our library10.1021/acs.est.8b04393

  38. 38.

    McKinney CW, Pruden A. Ultraviolet disinfection of antibiotic resistant bacteria and their antibiotic resistance genes in water and wastewater. Environ Sci Technol. 2012;46:13393-400.

    Not in our library10.1021/es303652q

  39. 39.

    Yoon Y, Chung HJ, Wen Di DY, Dodd MC, Hur HG, Lee Y. Inactivation efficiency of plasmid-encoded antibiotic resistance genes during water treatment with chlorine, UV, and UV/H2O2. Water Res. 2017;123:783-93.

    Not in our library10.1016/j.watres.2017.06.056

  40. 40.

    Rizzo L, Manaia C, Merlin C, Schwartz T, Dagot C, Ploy MC, et al. Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: a review. Sci Total Environ. 2013;447:345-60.

    Not in our library10.1016/j.scitotenv.2013.01.032

  41. 41.

    Roller VO SD, Kawata K. Mode of bacterial inactivation by chlorine dioxide. Water Res. 1980;14:635-41.

    Not in our library

  42. 42.

    Lim Y, Su CH, Liao YC, Lee SY. Impedimetric analysis on the mass transfer properties of intact and competent E. coli cells. Biochim Biophys Acta Biomembr. 2019;1861:9-16.

    Not in our library10.1016/j.bbamem.2018.10.010

  43. 43.

    Leclerc H, Schwartzbrod L, Dei-Cas E. Microbial agents associated with waterborne diseases. Crit Rev Microbiol. 2002;28:371-409.

    Not in our library10.1080/1040-840291046768

  44. 44.

    Zhang Y, Zhuang Y, Geng J, Ren H, Zhang Y, Ding L, et al. Inactivation of antibiotic resistance genes in municipal wastewater effluent by chlorination and sequential UV/chlorination disinfection. Sci Total Environ. 2015;512-513:125-32.

    Not in our library10.1016/j.scitotenv.2015.01.028

  45. 45.

    Seidman CE, Struhl K, Sheen J, Jessen T. Introduction of plasmid DNA into cells. Curr Protoc Mol Biol. 2001;37:1.8.1-1.8.10.

    Not in our library10.1002/0471142727.mb0108s37

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