Evaluation the Impact of Chlorination on the Development of Chlorine and Antibiotic-Resistant Bacterial Isolates in Public Swimming Pools

Water is necessary for human daily activities and acts as a vehicle for the transmission of several waterborne diseases. This study was designed to investigate the bacteriological safety of water in swimming pools in Sulaymaniyah city-Iraq. For this purpose, during summer time (August to September 2021), sixty-seven water samples were collected from eight indoor public swimming pools. The physicochemical parameters were measured and the bacteriological assessment was done using the Most Probable Number test. The diversity of microbial communities was then assessed using the Vitek 2 compact and the 16S rRNA gene. In addition, the chlorine resistance test and biofilm formation were quantified using the microdilution technique and crystal violet staining method, respectively. Moreover, the Kirby-Bauer disk diffusion technique was used to test antimicrobial susceptibility. Variable results of the physicochemical analysis were recorded for each of the temperatures (25.81±1.16 to 28.69±1.03 °C), pH (7.02±0.24 to 7.46±0.31), and free chlorine (0.12±0.05 to 1.38±1.24 mg L -1 ). MPN negative results revealed that non-lactose fermenting bacteria were detected in 80% of the water samples, including Enterobacter cloacae 34/59 (57.63%), and Pseudomonas spp. 10/59 (16.95%), and other important opportunistic pathogens 15/59 (25.42%). The contamination rate was strongly correlated with the sources of water used for the swimming pool and the free chlorine concentrations in the pools. Chlorine resistance test results revealed variations for all isolated bacteria ranging from 1.25 to 5 mg L -1 . Moreover, a negative linear correlation was found between chlorine concentrations, and both bacterial growth and biofilm formation. Most significantly, there was a strong positive correlation (r=0.90 to 0.98) between the degree of biofilm formation and bacterial growth. Moreover, different antibiogram profiles were recorded for the 15 antibiotics used in this study. It can be concluded that most of the studied swimming pools are heavily contaminated by chlorine and antibiotic-resistant waterborne pathogens. Therefore, continuous monitoring and use of maximum permissible chlorine disinfectant are highly recommended.

[1]  A. Olaniran,et al.  Does Chlorination Promote Antimicrobial Resistance in Waterborne Pathogens? Mechanistic Insight into Co-Resistance and Its Implication for Public Health , 2022, Antibiotics.

[2]  N. Desai,et al.  Identification and characterization of chlorine-resistant bacteria from water distribution sites of Mumbai , 2021, Archives of Microbiology.

[3]  L. M. Pitombo,et al.  Swimming pools disinfection by chlorine-based products in a simulated environment / Desinfecção de piscinas com produtos à base de cloro em ambiente simulado , 2021 .

[4]  Aifeng Li,et al.  Chlorine disinfectants promote microbial resistance in Pseudomonas sp. , 2021, Environmental research.

[5]  I. Gheorghe,et al.  Antibiotic resistance profiles in cultivable microbiota isolated from some romanian natural fishery lakes included in Natura 2000 network , 2021, BMC Veterinary Research.

[6]  T. Stenström,et al.  Detection of multidrug resistant environmental isolates of acinetobacter and Stenotrophomonas maltophilia: a possible threat for community acquired infections? , 2020, Journal of environmental science and health. Part A, Toxic/hazardous substances & environmental engineering.

[7]  Hong-Ying Hu,et al.  Evaluating method and potential risks of chlorine-resistant bacteria (CRB): A review. , 2020, Water research.

[8]  Zhiqiang Shen,et al.  Chlorine disinfection promotes the exchange of antibiotic resistance genes across bacterial genera by natural transformation , 2020, The ISME Journal.

[9]  Peili Lu,et al.  Granular activated carbon assisted nitrate-dependent anaerobic methane oxidation-membrane bioreactor: Strengthening effect and mechanisms. , 2020, Environment international.

[10]  Z. Ahmed,et al.  Evaluation of the efficacy of commonly used disinfectants against isolated chlorine-resistant strains from drinking water used in Egyptian cattle farms , 2019, Veterinary world.

[11]  M. R. Templeton,et al.  Chlorination and ultraviolet disinfection of antibiotic-resistant bacteria and antibiotic resistance genes in drinking water , 2019, AIMS Environmental Science.

[12]  H. Yesilkaya,et al.  Functional assessment of microbial superoxide dismutase isozymes suggests a differential role for each isozyme. , 2019, Free radical biology & medicine.

[13]  Minglu Zhang,et al.  Selective antibiotic resistance genes in multiphase samples during biofilm growth in a simulated drinking water distribution system: Occurrence, correlation and low-pressure ultraviolet removal. , 2019, The Science of the total environment.

[14]  A. Uhlemann,et al.  Multidrug-Resistant Enterobacter cloacae Complex Emerging as a Global, Diversifying Threat , 2019, Front. Microbiol..

[15]  H. Tang,et al.  Variability of residual chlorine in swimming pool water and determination of chlorine consumption for maintaining hygienic safety of bathers with a simple mass balance model. , 2018, Journal of water and health.

[16]  G. Kampf.  Biocidal Agents Used for Disinfection Can Enhance Antibiotic Resistance in Gram-Negative Species , 2018, Antibiotics.

[17]  C. Bertelli,et al.  Reduced Chlorine in Drinking Water Distribution Systems Impacts Bacterial Biodiversity in Biofilms , 2018, Front. Microbiol..

[18]  S. Monecke,et al.  Antimicrobial resistance in Enterobacteriaceae from healthy broilers in Egypt: emergence of colistin-resistant and extended-spectrum β-lactamase-producing Escherichia coli , 2018, Gut Pathogens.

[19]  Dong Yang,et al.  Chlorine disinfection increases both intracellular and extracellular antibiotic resistance genes in a full-scale wastewater treatment plant. , 2018, Water research.

[20]  Z. Teng,et al.  Association between bacterial survival and free chlorine concentration during commercial fresh-cut produce wash operation. , 2018, Food microbiology.

[21]  S. Saigal,et al.  Action and resistance mechanisms of antibiotics: A guide for clinicians , 2017, Journal of anaesthesiology, clinical pharmacology.

[22]  Xin Yu,et al.  Effect of sodium hypochlorite on typical biofilms formed in drinking water distribution systems. , 2017, Journal of water and health.

[23]  Amira Amine,et al.  Pseudomonas aeruginosa in swimming pools , 2017 .

[24]  M. A. Khan,et al.  Assessment of microbial quality in household water tanks in Dubai, United Arab Emirates , 2016 .

[25]  T. Grossman.  Tetracycline Antibiotics and Resistance. , 2016, Cold Spring Harbor perspectives in medicine.

[26]  W. Uhl,et al.  Rejection of submicron sized particles from swimming pool water by a monolithic SiC microfiltration membrane: Relevance of steric and electrostatic interactions , 2016 .

[27]  N. Khanjani,et al.  Survey of bacterial contamination of environment of swimming pools in Yazd city, in 2013 , 2015 .

[28]  Jian Yang,et al.  Fate of Antibiotic Resistant Bacteria and Genes during Wastewater Chlorination: Implication for Antibiotic Resistance Control , 2015, PloS one.

[29]  Stuart J. Khan,et al.  Chemical contaminants in swimming pools: Occurrence, implications and control. , 2015, Environment international.

[30]  Michael B. Fisher,et al.  Global Monitoring of Water Supply and Sanitation: History, Methods and Future Challenges , 2014, International journal of environmental research and public health.

[31]  Yuefeng F. Xie,et al.  Haloacetic acids in swimming pool and spa water in the United States and China , 2014, Frontiers of Environmental Science & Engineering.

[32]  T. Takaro,et al.  Recreational water-related illness: office management and prevention. , 2013, Canadian family physician Medecin de famille canadien.

[33]  S. Rice,et al.  A risk assessment of Pseudomonas aeruginosa in swimming pools: a review. , 2012, Journal of water and health.

[34]  M. M. Abd El-Salam.  Assessment of water quality of some swimming pools: a case study in Alexandria, Egypt , 2012, Environmental Monitoring and Assessment.

[35]  R. Briandet,et al.  Resistance of bacterial biofilms to disinfectants: a review , 2011, Biofouling.

[36]  Hong-Ying Hu,et al.  Inactivation and reactivation of antibiotic-resistant bacteria by chlorination in secondary effluents of a municipal wastewater treatment plant. , 2011, Water research.

[37]  F. Frimmel,et al.  Trichloramine in swimming pools--formation and mass transfer. , 2011, Water research.

[38]  J. Cabral,et al.  Water Microbiology. Bacterial Pathogens and Water , 2010, International journal of environmental research and public health.

[39]  H. Nelis,et al.  In vitro activity of ceftazidime, ciprofloxacin, meropenem, minocycline, tobramycin and trimethoprim/sulfamethoxazole against planktonic and sessile Burkholderia cepacia complex bacteria. , 2009, The Journal of antimicrobial chemotherapy.

[40]  A. Rabi,et al.  Sanitary Conditions of Public Swimming Pools in Amman, Jordan , 2008, International journal of environmental research and public health.

[41]  George Filioussis,et al.  Microbiological quality of indoor and outdoor swimming pools in Greece: investigation of the antibiotic resistance of the bacterial isolates. , 2008, International journal of hygiene and environmental health.

[42]  A. Rabi,et al.  Sanitary Conditions of Public Swimming Pools in Amman, Jordan , 2007, International journal of environmental research and public health.

[43]  M. Beach,et al.  Outbreak of norovirus illness associated with a swimming pool , 2006, Epidemiology and Infection.

[44]  Arto S. Baghdayan,et al.  Enterococcal Surface Protein, Esp, Enhances Biofilm Formation by Enterococcus faecalis , 2004, Infection and Immunity.

[45]  C. Gugliandolo,et al.  Distribution of potentially pathogenic bacteria as free living and plankton associated in a marine coastal zone , 2004, Journal of applied microbiology.

[46]  P. Seth,et al.  Suboptimal chlorine treatment of drinking water leads to selection of multidrug-resistant Pseudomonas aeruginosa. , 2004, Ecotoxicology and environmental safety.

[47]  P. Vandamme,et al.  Influence of taxonomic status on the in vitro antimicrobial susceptibility of the Burkholderia cepacia complex. , 2002, The Journal of antimicrobial chemotherapy.

[48]  Donald C. Lehman,et al.  Textbook of Diagnostic Microbiology , 1997 .

[49]  G. Oktay,et al.  [Bacteriological examination of paper money]. , 1992, Mikrobiyoloji bulteni.

[50]  C. Gerba,et al.  Disinfection of bacteria in water systems by using electrolytically generated copper:silver and reduced levels of free chlorine. , 1990, Canadian journal of microbiology.

[51]  R. S. Tobin,et al.  Effect of chlorination on antibiotic resistance profiles of sewage-related bacteria , 1984, Applied and environmental microbiology.

[52]  S. Alcock.  Acute otitis externa in divers working in the North Sea: a microbiological survey of seven saturation dives , 1977, Journal of Hygiene.

[53]  J. O. Irwin,et al.  The estimation of the bactericidal power of the blood , 1938, Epidemiology and Infection.

[54]  Hastyar Najmuldeen.  Assessment of Chlorine Resistance Enterobacter cloacae Isolated from Water Storage Tanks in Sulaimaniyah City-Iraq , 2020 .

[55]  I. Karunasagar,et al.  Molecular Methods to Study Vibrio parahaemolyticus and Vibrio vulnificus From Atypical Environments , 2018 .

[56]  Derara Chalchisa,et al.  Assessment of the quality of drinking water in storage tanks and its implication on the safety of urban water supply in developing countries , 2017, Environmental Systems Research.

[57]  Joyce Margaret Ekopai,et al.  Determination of Bacterial Quality of Water in Randomly Selected Swimming Pools in Kampala City, Uganda , 2017, Adv. Artif. Neural Syst..

[58]  H. Horn,et al.  Occurrence and simulation of trihalomethanes in swimming pool water: A simple prediction method based on DOC and mass balance. , 2016, Water research.

[59]  C. Saba,et al.  Water Quality Assessment of Swimming Pools and Risk of Spreading Infections in Ghana , 2015 .

[60]  Microbial Qualities of Swimming Pools in Lagos, Nigeria , 2013 .

[61]  Challenges in global immunization and the Global Immunization Vision and Strategy 2006-2015. , 2006, Releve epidemiologique hebdomadaire.

[62]  Mark D. Sobsey,et al.  Managing Water in the Home: Accelerated Health Gains from Improved Water Supply , 2002 .

[63]  M. Ferraro.  Performance standards for antimicrobial susceptibility testing , 2001 .

[64]  J. Czeczelewski.  [Evaluation of water sanitation and hygiene in swimming pools in the town of Biała Podlaska]. , 1994, Roczniki Panstwowego Zakladu Higieny.

[65]  A. Bauer,et al.  Antibiotic susceptibility testing by a standardized single disk method. , 1966, Technical bulletin of the Registry of Medical Technologists. American Society of Clinical Pathologists. Registry of Medical Technologists.