Assessment of Chlorine Resistance Enterobacter cloacae Isolated from Water Storage Tanks in Sulaimaniyah City-Iraq

Document Type : Original Article

Author

Department of biology/ college of science/ university of Sulaimani

Abstract

This study was designed to determine the extent of contamination of water storage tanks by non-lactose fermenter Enterobacter spp, and to characterize the chlorine and antibiotic resistance status. Moreover, to find the correlation between biofilm formation and resistance to chlorine. For this purpose, a total of 60 water samples were collected from residential and restaurant water storage tanks. Bacterial analysis and antibiotic susceptibility profiles of the samples were assessed by the Most Probable Number (MPN) and Vitek 2 compact tests, respectively. The biofilm formation was quantified by crystal violet staining method and chlorine resistance test by microdilution technique.
Obtained results indicated that water samples were contaminated by Escherichia coli (35%). Additionally, water samples that were assessed to be potable by the MPN test showed that (44%) positive for Enterobacter cloacae. Results of chlorine resistance test revealed variation in resistance of E. cloacae to different concentrations of chlorine, and relatively similar antibiotic susceptibility profiles. Moreover, biofilm analysis showed the isolates that were resistant to concentration of chlorine 400 mg L-1, have formed significantly more biofilm than those that were resistant to other concentrations. A positive non-linear correlation (r = 0.72) was found between the degree of biofilm formation and the ability of isolates to resist different chlorine concentrations, and no correlation has been detected between antibiotic and chlorine resistance.
It can be concluded that the presence of chlorine resistant E. cloacae in drinking water can pose a real public health threat. The routine microbial water analysis should be modified to include detection of non-lactosefermenter Enterobacter.

Keywords

  1. References

    1. KHAN, M. A., & ALMADANI, A. M. A. 2017. Assessment of microbial quality in household water tanks in Dubai, United Arab Emirates. Environ. Eng. Res. 22(1): 55-60.
    2. FARKAS, A., BOCOȘ, B., BULARDA, M. D. & CRACIUNAS, C. 2014. Effect of different disinfectants against biofilm bacteria. Studia Universitatis Babes-Bolyai Biologia, LIX. 1: 5-20.
    3. LEVY, K., ANDERSON, L., ROBB, K. A., CEVALLOS, W., TRUEBA, G. & EISENBERG J. N. 2014. Household effectiveness vs. laboratory efficacy of point-of-use chlorination. Water Res. 54:69-77.
    4. SHRIVASTAVA, R., UPRETI, R. K., JAIN, S. R., ORASAD K. N., SETH, P. K. & CHATURVEDI, U. C. 2004. Suboptimal chlorine treatment of drinking water leads to selection of multidrug-resistant Pseudomonas aeruginosa. Ecotoxicol. Environ. Saf. 58(2): 277-283.
    5. SUN, W., LIU, W., CUI, L., ZHANG, M. & WANG, B. 2013. Characterization and identification of a chlorine-resistant bacterium, Sphingomonas TS001, from a model drinking water distribution system. Sci. Total Environ. 458-460:169-175.
    6. DESTIANI, R. & TEMPLETON, M. R. 2019. Chlorination and ultraviolet disinfection of antibiotic-resistant bacteria and antibiotic resistance genes in drinking water. AIMS Environmental Science. 6(3):222-241.
    7. KAMAL, M. A., KHALAF, M. A., AHMED, Z. & JAKEE, J. E. 2019. Evaluation of the efficacy of commonly used disinfectants against isolated chlorine-resistant strains from drinking water used in Egyptian cattle farms. Vet. World. 12(12): 2025-2035.
    8. SANGANYADO, E. & GWENZI, W. 2019. Antibiotic resistance in drinking water systems: Occurrence, removal, and human health risks. Sci Total Environ. 669: 785-797.
    9. AKUFFO, I., COBBINA, S. J., ALHASSAN, E. H. & NKOOM, M. 2013. Assessment of the quality of water before and after storage in the Nyankpala community of the Tolon-Kumbungu District, Ghana. IJSTR. 2(2):10-13.
    10. BRICK, T., PRIMROSE, B., CHANDRASEKHAR, R., ROY, S., MULIYIL, J., & KANG, G. 2004. Water contamination in urban south India: household storage practices and their implications for water safety and enteric infections. Int J Hyg Environ Health.5: 473-480.
    11. CHALCHISA, D. MEGERSA, M. & BEYENE, A. 2018. Assessment of the quality of drinking water in storage tanks and its implication on the safety of urban water supply in developing countries. Environ Syst Res. 6(12):1-6.
    12. KAMPF, G. 2018. Biocidal agents used for disinfection can enhance antibiotic resistance in gram-negative species. Antibiotics. 7(4):1-24.
    13. ZHU, Z., WU, C., ZHONG, D., YUAN, Y., SHAN, L. & ZHANG, J. 2014. Effects of pipe materials on chlorine-resistant biofilm formation under long-term high chlorine level. Applied biochem Biotechnol.173 (6): 1-16.
    14. BERTELLI, C., COURTOIS, S., ROSIKIEWICZ, M. & et al. 2018. Reduced chlorine in drinking water distribution systems impacts bacterial biodiversity in biofilms. Front Microbiol. 9:1-11.
    15. BRIDIER, A., BRIANDET, R., THOMAS, V. & DUBOIS-BIOFOULING, F. 2011. Resistance of bacterial biofilms to disinfectants: a review. Biofouling. 27(9):1017-1032.
    16. CABRAL, J. P. S. 2010. Water microbiology. Bacterial pathogens and water. Int. J. Environ. Res. Public Health. 7: 3657-3703.
    17. KANAMORI, H., WEBER, D. J. & RUTALA, W. A. 2016. Healthcare Outbreaks Associated with a Water Reservoir and Infection Prevention Strategies. Clin. Infect. Dis. 62(11):1423-1435.
    18. DAVIN-REGLI, A. & PAGÈS, J.-M. 2015. Enterobacter aerogenes and Enterobacter cloacae; versatile bacterial pathogens confronting antibiotic treatment. Front Microbiol. 6:1-10.
    19. MEZZATESTA, M. L., GONA, F. & STEFANI, S. 2012. Enterobacter cloacae complex: clinical impact and emerging antibiotic resistance. Future microbiol. 7(7): 887-902.
    20. TALARO, K. P. and TALARO, A., 2002. Foundations in Microbiology, 4th edit. The McGraw−Hill Companies. 810-811.
    21. SAPUTRO, S., YOSHIMURA, K., TAKEHARA, K., MATSUOKA, S., NARSITO. 2011. Analytical methods of chlorine and the substances produced by the chlorine treatments, in: Chlorine Properties. Applications and Health Effects. pp. 259–274.
    22. World Health Organization WHO, 2006. Guideline for drinking-water quality: incorporating first addendum recommendations, 3rd ed. World Health Organization, Geneva.
    23. BAUER, A.W., KIRBY, W.M., SHERRIS, J.C., TURCK, M., 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol. 45:493–496.
    24. MAHON, C. and MANUSELIS, G. 2000. Text book of diagnostic microbiology. 2nd edition. W.B. Saunders Company.
    25. CLSI, 2013. Performance Standards for Antimicrobial Susceptibility Testing; Clinical and Laboratory Standards Institute. https://doi.org/1-56238-525-5.
    26. HUANG, J. J., HU, H. Y., TANG, F., LI, Y., LU, S. Q., & LU, Y. 2011. Inactivation and reactivation of antibiotic-resistant bacteria by chlorination in secondary effluents of a municipal wastewater treatment plant. Water research. 45(9): 2775-2781.
    27. YUAN, Q. B., GUO, M. T. & YANG, J. 2015. Fate of antibiotic resistant bacteria and genes during wastewater chlorination: implication for antibiotic resistance control. PloS one. 1-11.
    28. TENDOLKAR, P. M., BAGHDAYAN, A. S., GILMORE, M. S. & SHANKAR, N. 2004. Enterococcal Surface Protein, Esp, Enhances Biofilm Formation by Enterococcus faecalis. Infect. Immun, 72(10):6032-6039.
    29. SUO, Y., HUANG, Y., LIU, Y., SHI, C. & SHI, X. 2012. The expression of superoxide dismutase (SOD) and a putative ABC transporter permease is inversely correlated during biofilm formation in Listeria monocytogenes 4b G. PLoS One. 7(10):1-9.
    30. NAJMULDEEN, H., ALGHAMDI, R., ALGHOFAILI, F., YESILKAYA, H. 2019. Functional assessment of microbial superoxide dismutase isozymes suggests a differential role for each isozyme. Free Radic. Biol. Med. 134, 215–228.
    31. World Health Organization WHO, 2014. Conflict and humanitarian crisis in Iraq, public health risk assessment and intervention.
    32. ALWAN, A. 2004. Health in Iraq. Minister of Health, Iraq.1-80.
    33. MUYIMA, N.Y.O., NGCAKANI , F. 1998. Indicator bacteria and regrowth potential of the drinking water in Alice, Eastern Cape. Water SA. 24: 29–34.
    34. TABOR, M., KIBRET, M., ABER, B. 2011. Bacteriological and Physicochemical Quality of Drinking Water and Hygiene- Sanitation Practices of the Consumers in Bahir Dar City, Ethiopia. Ethiop. J. Health Sci. 21.(1):19-26.
    35. TOKAJIAN, S. & HASHWA, F. 2004. Microbiological quality and genotypic speciation of heterotrophic bacteria isolated from potable water stored in household tanks. Water Quality Research Journal. 39(1): 64-73.
    36. ALOBAIDY, A.H.M.J., ABID, H.S., MAULOOD, B.K. 2010. Application of Water Quality Index for Assessment of Dokan Lake Ecosystem, Kurdistan Region, Iraq. J. Water Resour. Prot. 02, 792–798.
    37. SALIH, F. A., OTHMAN, N., MUHIDIN, F. M. and KASEM, A. O. 2015. Assessment of the Quality of Water in Sulaimaniyah City, Kurdistan Region: Iraq.Curr. World Environ. 10(3): 781-791.
    38. JIN, M., LIU, L., WANG, D. NING, YANG, D., LIU, W. LI, YIN, J., YANG, Z. WEI, WANG, H. RAN, QIU, Z. GANG, SHEN, Z. QIANG, SHI, D. YANG, LI, H. BEI, GUO, J. HUA, LI, J. WEN. 2020. Chlorine disinfection promotes the exchange of antibiotic resistance genes across bacterial genera by natural transformation. ISME J. 14 (7): 1847–1856.
    39. LEONG, L. Y. C., OTSAKA, D. & RIDGWAY, H. F. 1982. Chlorine-Resistance of Coliform-Tested Bacteria Isolated from Raw and Treated Sewage Effluents. Water Sci Tecnol. 14(4-5): 127-132.
    40. LUO, L., WU, Y., YU, T., WANG, Y., CHEN, G., TONG, X., BAI, Y., XU, C., WANG, H., IKUNO, N., HU, H. (2020). Evaluating method and potential risks of chlorine-resistant bacteria (CRB): A review. Water Research, 188.
    41. AL-BERFKANI, M. I., ZUBAIR, A. I. & BAYAZED, H. 2014. Assessment of chlorine resistant bacteria and their susceptibility to antibiotic from water distribution system in Duhok province. J. appl. biol. biotechnol.2(6):10-13.
    42. RIDGWAY, H. F. & OLSON, B. H. 1982. Chlorine resistance patterns of bacteria from two drinking water distribution systems. Appl. Environ. Microbiol. 44(4): 972-987.
    43. SIMÕES, L., SIMÕES, M. & VIEIRA, M. 2010. Influence of the Diversity of Bacterial Isolates from Drinking Water on Resistance of Biofilms to Disinfection. AEM. 76(19): 6673-6679.
    44. AL-BAHRY, S. N., AL-HINAI, J. A. & PROCEDIA, M.-I. Y. 2013. Opportunistic and microbial pathogens in municipal water distribution systems. APCBEE procedia. 5: 339-343.
    45. MEDRANO‐FÉLIX, A., MARTINEZ, C., CASTRO-DEL, C.N, LEON-FELIX, J, PERAZA-GARAY, F. GERBA, C. P. & CHAIDEZ, C. 2011. Impact of prescribed cleaning and disinfectant use on microbial contamination in the home. J Appl. Microbiol. 110(2): 463-470.
    46. GROBE, S., WINGENDER, J. & FLEMMING, H-C. 2001. Capability of mucoid Pseudomonas aeruginosa to survive in chlorinated water. IJHEH. 204(2-3): 139-142.
    47. LIN, H., ZHU, X., WANG, Y., YU, X. 2017. Effect of sodium hypochlorite on typical biofilms formed in drinking water distribution systems. JWH. 218-227.
    48. PENNA, V., MARTINS, S. & MAZZOLA, P. 2002. Identification of bacteria in drinking and purified water during the monitoring of a typical water purification system. BMC Public Health. 2(13):1-11.
    49. CAI, L., WANG, H., LIANG, L., WANG, G., Xu, X., and WANG, H. 2018. Response of Formed‐Biofilm of Enterobacter cloacae, Klebsiella oxytoca, and Citrobacter freundii to Chlorite‐Based Disinfectants. J food science. 83 (5): 1326-1332.
    50. BUSH, K., BRADFORD, P.A., 2016. β-lactams and β-lactamase inhibitors: An overview. Cold Spring Harb. Perspect. Med. 6 (8): 1-22.
    51. KAPOOR, G, SAIGAL, S., ELONGAVAN, A. 2017. Action and resistance mechanisms of antibiotics: A guide for clinicians. J Anaesthesiol Clin Pharmacol. 33(3):300-305.
    52. ANNAVAJHAL, M.K., GOMEZ-SIMMONDS, A., Uhlemann, A.C. 2019. Multidrug-resistant Enterobacter cloacae complex emerging as a global, diversifying threat. Front. Microbiol. 10(44): 1-8.
    53. BASSETTI, M., GINOCCHIO, F., MIKULSKA, M. 2011. New treatment options against gram-negative organisms. Crit. Care. 15:1-9.
    54. GROSSMAN, T.H., 2016. Tetracycline antibiotics and resistance. Cold Spring Harb. Perspect. Med. 6(4):a025387.
    55. KEENEY, D., RUZIN, A., BRADFORD, P.A. 2007. RamA, a transcriptional regulator, and AcrAB, an RND-type efflux pump, are associated with decreased susceptibility to tigecycline in Enterobacter cloacae. Microb. Drug Resist. 13(1):1–6.
    56. KIM, S.Y., Park, Y.J., Yu, J.K., Kim, Y.S., 2011. Aminoglycoside susceptibility profiles of enterobacter cloacae isolates harboring the aac(6′)-Ib gene. Korean J. Lab. Med. 31: 279–281.
    57. MOAWAD, A.A., HOTZEL, H., NEUBAUER, H., EHRICHT, R., MONECKE, S., TOMASO, H., HAFEZ, H.M., ROESLER, U., El-ADAWY, H.. 2018. Antimicrobial resistance in Enterobacteriaceae from healthy broilers in Egypt: Emergence of colistin-resistant and extended-spectrum β-lactamase-producing Escherichia coli. Medical Mi. Gut Pathog. 10(39):1-12.
    58. RAMOS, M.M.B., GAETTI-JARDIM, E.C., GAETTI-JARDIM, E. 2009. Resistance to tetracycline and β-lactams and distribution of resistance markers in enteric microorganisms and pseudomonads isolated from the oral cavity. J. Appl. Oral Sci. 17:13–18.
    59. TANNER, W.D., VANDERSLICE, J.A., GOEL, R.K., LEECASTER, M.K., FISHER, M.A., OLASTADT, J., GURLEY, C.M., MORRIS, A.G., SEELY, K.A., CHAPMAN, L., KORANDO, M., SHABAZZ, K.A., STADSHOLT, A., VANDEVLDE, J., BRAUN-HOWLAND, E., MINIHANE, C., HIGGINS, P.J., DERAS, M., JABER, O., JETTE, D., GUNDLAPALLI, A. V. 2019. Multi-state study of Enterobacteriaceae harboring extended-spectrum beta-lactamase and carbapenemase genes in U.S. drinking water. Sci. Rep. 9. 3938.
    60. AlTURAIFI, F.H., ALMOMATIN, A.A., BADGER-EMEKA, L., EMEKA, P.M., ISLAM, M.M. 2019. Assessment of Microbiological Content of Private and Public Recreational Water Facilities and Their Antimicrobial Susceptibility Pattern in Al-Ahsa. Environ. Health Insights. 13:1-18.
    61. MARTINEZ, J.L. 2009. Environmental pollution by antibiotics and by antibiotic resistance determinants. Environ. Pollut. 157(11):2893-902.
    62. RTEDEL, S., BORIRE, N., CARSON, K.A., VADLMUDI, A., KHUVIS, J., VADLAMUDI, V., ATUKORALE, V., RIEDEL, V.A.A., PARRISH, N.M. 2019. A survey of antimicrobial resistance in Enterobacteriaceae isolated from the Chesapeake Bay and adjacent upper tributaries. Microbiologyopen. 8:1-14.
    63. HUANG, J. J., HU, H., LU, S. Q., LI, Y., TANG, F. & WEI, B. 2012. Monitoring and evaluation of antibiotic-resistant bacteria at a municipal wastewater treatment plant in China. Environ Int. 42:31-36.
    64. KORZENIEWSKA, E., KORZENIEWSKA, A. & HARNISZ, M. 2013. Antibiotic resistant Escherichia coli in hospital and municipal sewage and their emission to the environment. Ecotoxicol. Environ. Saf. 91: 96-102.
    65. MURRAY, G. E., TOBIN, R. S., JUNKINS, B., & KUSHNER, D. J. 1984. Effect of chlorination on antibiotic resistance profiles of sewage-related bacteria. AEM. 48(1): 73-77.
    66. IWANE, T., URASE, T. & YAMAMOTO, K. 2001. Possible impact of treated wastewater discharge on incidence of antibiotic resistant bacteria in river water. Water Sci Technol. 43(2): 91-99.
    67. RIZZO, L., MANAIA, C., MERLIN, C., SCHWARTZ, T., DAGOT, C., PLOY, M. C., MICHAEL, I. & FATTA-KASSINOS, D. 2013. Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment: A review. Sci Total Environ. 447: 345-360.