Isolation of Environmental Bacteria from Surface and Drinking Water in Mafikeng, South Africa, and Characterization Using Their Antibiotic Resistance Profiles
The aim of this study was to isolate and identify environmental bacteria from various raw water sources as well as the drinking water distributions system in Mafikeng, South Africa, and to determine their antibiotic resistance profiles. Water samples from five different sites (raw and drinking water) were analysed for the presence of faecal indicator bacteria as well as Aeromonas and Pseudomonas species. Faecal and total coliforms were detected in summer in the treated water samples from the Modimola dam and in the mixed water samples, with Pseudomonas spp. being the most prevalent organism. The most prevalent multiple antibiotic resistance phenotype observed was KF-AP-C-E-OT-K-TM-A. All organisms tested were resistant to erythromycin, trimethoprim, and amoxicillin. All isolates were susceptible to ciprofloxacin and faecal coliforms and Pseudomonas spp. to neomycin and streptomycin. Cluster analysis based on inhibition zone diameter data suggests that the isolates had similar chemical exposure histories. Isolates were identified using gyrB, toxA, ecfX, aerA, and hylH gene fragments and gyrB, ecfX, and hylH fragments were amplified. These results demonstrate that (i) the drinking water from Mafikeng contains various bacterial species and at times faecal and total coliforms. (ii) The various bacteria are resistant to various classes of antibiotics. 1. Introduction Water is considered a vehicle for the propagation and dissemination of human associated bacteria [1]. Safe drinking water is a fundamental human right and if contaminated with opportunistic pathogenic environmental bacteria, it may have health implications for consumers [2, 3]. Human health should therefore be protected by preventing microbial contamination of water that is intended for consumption [4]. In rural communities, untreated surface water from rivers, dams, and streams is directly used for drinking and other domestic purposes [5]. These unprotected water sources can be contaminated with microbes through rainfall run-off and agricultural inputs, mixing with sewage effluents and faeces from wild life [6, 7], which render them unacceptable for human consumption. Faecal coliforms, Aeromonas and Pseudomonas, are used as indicators of faecal contamination in water [8] and the presence of these pathogens may have severe health implications on consumers especially those that are immunocompromised [5, 9, 10]. South Africa is a semiarid country with very low rainfall and high evaporation [11] and it has a scarcity of fresh water systems due to the highly variable and spatial distribution of
References
[1]
C. Faria, I. Vaz-Moreira, E. Serapicos, O. C. Nunes, and C. M. Manaia, “Antibiotic resistance in coagulase negative staphylococci isolated from wastewater and drinking water,” Science of the Total Environment, vol. 407, no. 12, pp. 3876–3882, 2009.
[2]
WHO, Guidelines for Drinking Water Quality, vol. 1, World Health Organization, Geneva, Switzerland, 3rd edition, 2004.
[3]
J. Fawell and M. J. Nieuwenhuijsen, “Contaminants in drinking water Environmental pollution and health,” British Medical Bulletin, vol. 68, no. 1, pp. 199–208, 2003.
[4]
S. V？lker, C. Schreiber, and T. Kistemann, “Drinking water quality in household supply infrastructure—a survey of the current situation in Germany,” International Journal of Hygiene and Environmental Health, vol. 213, no. 3, pp. 204–209, 2010.
[5]
P. T. Biyela, J. Lin, and C. C. Bezuidenhout, “The role of aquatic ecosystems as reservoirs of antibiotic resistant bacteria and antibiotic resistance genes,” Water Science and Technology, vol. 50, no. 1, pp. 45–50, 2004.
[6]
C. L. Obi, N. Potgieter, P. O. Bessong, and G. Matsaung, “Assessment of the microbial quality of river water sources in rural Venda communities in South Africa,” Water SA, vol. 28, no. 3, pp. 287–292, 2002.
[7]
A. Sharma, N. Dubey, and B. Sharan, “Characterization of aeromonads isolated from the river Narmada, India,” International Journal of Hygiene and Environmental Health, vol. 208, no. 5, pp. 425–433, 2005.
[8]
L. F. Webster, B. C. Thompson, M. H. Fulton et al., “Identification of sources of Escherichia coli in South Carolina estuaries using antibiotic resistance analysis,” Journal of Experimental Marine Biology and Ecology, vol. 298, no. 2, pp. 179–195, 2004.
[9]
S. Dumontet, K. Krovacek, S. B. Svenson, V. Pasquale, S. B. Baloda, and G. Figliuolo, “Prevalence and diversity of Aeromonas and Vibrio spp. in coastal waters of Southern Italy,” Comparative Immunology, Microbiology and Infectious Diseases, vol. 23, no. 1, pp. 53–72, 2000.
[10]
D. Pavlov, C. M. E. de Wet, W. O. K. Grabow, and M. M. Ehlers, “Potentially pathogenic features of heterotrophic plate count bacteria isolated from treated and untreated drinking water,” International Journal of Food Microbiology, vol. 92, no. 3, pp. 275–287, 2004.
[11]
R. Eberhard and P. Robinson, Guidelines for the Development of National Water Policies and Strategies to Support IWRM 2003 Draft, SADC Water Sector Co-Ordination Unit, Gaborone, Botswana, 2003.
[12]
J. R. Adewumi, A. A. Ilemobade, and J. E. Van Zyl, “Treated wastewater reuse in South Africa: overview, potential and challenges,” Resources, Conservation and Recycling, vol. 55, no. 2, pp. 221–231, 2010.
[13]
International Water Association (IWA), 2008.
[14]
R. Mckenzie, J. Buckle, W. Wegelin, and N. Meyer, “Water demand management cookbook, rand water in collaboration with WRP, managing water for African Cities UN Habitat Programme, New York,” Medical Bulletin, vol. 68, pp. 199–208, 2003.
[15]
S. Dimitriadis, Issues Encountered in Advancing Australia's Water Recycling Schemes, Research Brief, Parliamentary Library, Parliament of Australia, Department of Parliamentary Services, 2005.
[16]
B. Vaseeharan, P. Ramasamy, T. Murugan, and J. C. Chen, “In vitro susceptibility of antibiotics against Vibrio spp. and Aeromonas spp. isolated from Penaeus monodon hatcheries and ponds,” International Journal of Antimicrobial Agents, vol. 26, no. 4, pp. 285–291, 2005.
[17]
J. L. Martinez, “Environmental pollution by antibiotics and by antibiotic resistance determinants,” Environmental Pollution, vol. 157, no. 11, pp. 2893–2902, 2009.
[18]
M. Akram, M. Shahid, and A. U. Khan, “Etiology and antibiotic resistance patterns of community-acquired urinary tract infections in J N M C Hospital Aligarh, India,” Annals of Clinical Microbiology and Antimicrobials, vol. 6, article 4, pp. 1–10, 2007.
[19]
A. ？uczkiewicz, K. Jankowska, S. Fudala-Ksiazek, and K. Olańczuk-Neyman, “Antimicrobial resistance of fecal indicators in municipal wastewater treatment plant,” Water Research, vol. 44, no. 17, pp. 5089–5097, 2010.
[20]
J. E. Moore, P. J. A. Moore, B. C. Millar et al., “The presence of antibiotic resistant bacteria along the River Lagan,” Agricultural Water Management, vol. 98, no. 1, pp. 217–221, 2010.
[21]
T. Schwartz, W. Kohnen, B. Jansen, and U. Obst, “Detection of antibiotic-resistant bacteria and their resistance genes in wastewater, surface water, and drinking water biofilms,” FEMS Microbiology Ecology, vol. 43, no. 3, pp. 325–335, 2003.
[22]
T. I. Lobova, Y. V. Barkhatov, O. V. Salamatina, and L. Y. Popova, “Multiple antibiotic resistance of heterotrophic bacteria in the littoral zone of Lake Shira as an indicator of human impact on the ecosystem,” Microbiological Research, vol. 163, no. 2, pp. 152–160, 2008.
[23]
Y. Zhang, C. F. Marrs, C. Simon, and C. Xi, “Wastewater treatment contributes to selective increase of antibiotic resistance among Acinetobacter spp.,” Science of the Total Environment, vol. 407, no. 12, pp. 3702–3706, 2009.
[24]
S. G. Mulamattathil, H. A. Esterhuysen, and P. J. Pretorius, “Antibiotic-resistant Gram-negative bacteria in a virtually closed water reticulation system,” Journal of Applied Microbiology, vol. 88, no. 6, pp. 930–937, 2000.
[25]
J. Lin and P. T. Biyela, “Convergent acquisition of antibiotic resistance determinants amongst the Enterobacteriaceae isolates of the Mhlathuze River, KwaZulu-Natal (RSA),” Water SA, vol. 31, no. 2, pp. 257–260, 2005.
[26]
C. W. Kinge and M. Mbewe, “Characterisation of Shigella species isolated from river catchments in the North West province of South Africa,” South African Journal of Science, vol. 106, no. 11-12, article 211, 2010.
[27]
SANS: South African National Standards, “Drinking water—part 1: microbiological , physical, chemical, aesthetic and chemical determinands,” SABS Standards Division, Pretoria, South Africa, 2011.
[28]
W. Kirby, W. Bauer, J. Sherris, and M. Turck, “Antibiotic susceptibility testing by a standardized single disk method,” The American Journal of Clinical Pathology, vol. 45, no. 4, pp. 493–496, 1966.
[29]
C. N. Ateba, M. Mbewe, and C. C. Bezuidenhout, “Prevalence of Escherichia coli O157 strains in cattle, pigs and humans in North West province, South Africa,” South African Journal of Science, vol. 104, no. 1-2, pp. 7–8, 2008.
[30]
National Committee for Clinical Laboratory Standards, “Performance standards for antimicrobial disk and dilution susceptibility tests for bacterial isolates from animals,” National Committee for Clinical Laboratory Standards, Wayne, Pa, USA, Approved standard, M31-A19, (11), 1999.
[31]
X. Qin, J. Emerson, J. Stapp, L. Stapp, P. Abe, and J. L. Burns, “Use of real-time PCR with multiple targets to identify Pseudomonas aeruginosa and other nonfermenting gram-negative bacilli from patients with cystic fibrosis,” Journal of Clinical Microbiology, vol. 41, no. 9, pp. 4312–4317, 2003.
[32]
A. A. Khan and C. E. Cerniglia, “Detection of Pseudomonas aeruginosa from clinical and environmental samples by amplification of the exotoxin A gene using PCR,” Applied and Environmental Microbiology, vol. 60, no. 10, pp. 3739–3745, 1994.
[33]
R. Lavenir, D. Jocktane, F. Laurent, S. Nazaret, and B. Cournoyer, “Improved reliability of Pseudomonas aeruginosa PCR detection by the use of the species-specific ecfX gene target,” Journal of Microbiological Methods, vol. 70, no. 1, pp. 20–29, 2007.
[34]
E. Kaszab, S. Szoboszlay, C. Dobolyi, J. Háhn, N. Pék, and B. Kriszt, “Antibiotic resistance profiles and virulence markers of Pseudomonas aeruginosa strains isolated from composts,” Bioresource Technology, vol. 102, no. 2, pp. 1543–1548, 2011.
[35]
A. H. Yousr, S. Napis, G. R. A. Rusul, and R. Son, “Detection of aerolysin and hemolysin genes in Aeromonas spp. isolated from environmental and shellfish sources by polymerase chain reaction,” ASEAN Food Journal, vol. 14, no. 2, pp. 115–122, 2007.
[36]
A. I. Sch？fer, H. M. A. Rossiter, P. A. Owusu, B. S. Richards, and E. Awuah, “Physico-chemical water quality in Ghana: prospects for water supply technology implementation,” Desalination, vol. 248, no. 1–3, pp. 193–203, 2009.
[37]
P. T. Yillia, N. Kreuzinger, and J. M. Mathooko, “The effect of in-stream activities on the Njoro River, Kenya. Part I: stream flow and chemical water quality,” Physics and Chemistry of the Earth, vol. 33, no. 8–13, pp. 722–728, 2008.
[38]
H. Schraft and L. A. Watterworth, “Enumeration of heterotrophs, fecal coliforms and Escherichia coli in water: comparison of 3M Petrifilm plates with standard plating procedures,” Journal of Microbiological Methods, vol. 60, no. 3, pp. 335–342, 2005.
[39]
W. Germs, M. S. Coetzee, L. van Rensburg, and M. S. Maboeta, “A preliminary assessment of the chemical and microbial water quality of the Chunies River—Limpopo,” Water SA, vol. 30, no. 2, pp. 267–272, 2004.
[40]
J. T. Carter, E. W. Rice, S. G. Buchberger, and Y. Lee, “Relationships between levels of heterotrophic bacteria and water quality parameters in a drinking water distribution system,” Water Research, vol. 34, no. 5, pp. 1495–1502, 2000.
[41]
C. L. Obi, P. O. Bessong, M. Momba, N. Potgieter, A. Samie, and E. O. Igumbor, “Profiles of antibiotic susceptibilities of bacterial isolates and physico-chemical quality of water supply in rural Venda communities, South Africa,” Water SA, vol. 30, no. 4, pp. 515–519, 2004.
[42]
P. Messi, E. Guerrieri, and M. Bondi, “Antibiotic resistance and antibacterial activity in heterotrophic bacteria of mineral water origin,” Science of the Total Environment, vol. 346, no. 1–3, pp. 213–219, 2005.
[43]
J. Lin, P. T. Biyela, and T. Puckree, “Antibiotic resistance profiles of environmental isolates from Mhlathuze River, KwaZulu-Natal (RSA),” Water SA, vol. 30, no. 1, pp. 23–28, 2004.
[44]
G. J. Oudhuis, A. Verbon, J. A. A. Hoogkamp-Korstanje, and E. E. Stobberingh, “Antimicrobial resistance in Escherichia coli and Pseudomonas aeruginosa from Intensive Care Units in The Netherlands, 1998–2005,” International Journal of Antimicrobial Agents, vol. 31, no. 1, pp. 58–63, 2008.
[45]
T. C. Siewicki, T. Pullaro, W. Pan, S. McDaniel, R. Glenn, and J. Stewart, “Models of total and presumed wildlife sources of fecal coliform bacteria in coastal ponds,” Journal of Environmental Management, vol. 82, no. 1, pp. 120–132, 2007.
[46]
A. Sorlózano, J. Gutiérrez, J. Luna et al., “High presence of extended-spectrum β-lactamases and resistance to quinolones in clinical isolates of Escherichia coli,” Microbiological Research, vol. 162, no. 4, pp. 347–354, 2007.
[47]
M. E. Scott, A. R. Melton-Celsa, and A. D. O'Brien, “Mutations in hns reduce the adherence of Shiga toxin-producing E. coli 091:H21 strain B2F1 to human colonic epithelial cells and increase the production of hemolysin,” Microbial Pathogenesis, vol. 34, no. 3, pp. 155–159, 2003.
[48]
E. Tuméo, H. Gbaguidi-Haore, I. Patry, X. Bertrand, M. Thouverez, and D. Talon, “Are antibiotic-resistant Pseudomonas aeruginosa isolated from hospitalised patients recovered in the hospital effluents?” International Journal of Hygiene and Environmental Health, vol. 211, no. 1-2, pp. 200–204, 2008.
[49]
E. Breidenstein, C. de la Fuente-Nú？ez, and R. Hancock, “Pseudomonas aeruginosa: all roads lead to resistance,” Trends in Microbiology, vol. 19, no. 8, pp. 419–426, 2011.
[50]
M. I. Jeena, P. Deepa, K. M. Mujeeb Rahiman, R. T. Shanthi, and A. A. M. Hatha, “Risk assessment of heterotrophic bacteria from bottled drinking water sold in Indian markets,” International Journal of Hygiene and Environmental Health, vol. 209, no. 2, pp. 191–196, 2006.
[51]
G. Emekdas, G. Aslan, S. Tezcan et al., “Detection of the frequency, antimicrobial susceptibility, and genotypic discrimination of Aeromonas strains isolated from municipally treated tap water samples by cultivation and AP-PCR,” International Journal of Food Microbiology, vol. 107, no. 3, pp. 310–314, 2006.
[52]
E. E. Odjadjare, E. O. Igbinosa, R. Mordi, B. Igere, C. L. Igeleke, and A. I. Okoh, “Prevalence of multiple antibiotics resistant (MAR) Pseudomonas species in the final effluents of three municipal wastewater treatment facilities in South Africa,” International Journal of Environmental Research and Public Health, vol. 9, no. 6, pp. 2092–2107, 2012.
[53]
S. Harakeh, H. Yassine, and E. Mutasem, “Antimicrobial-resistance patterns of Escherichia coli and Salmonella strains in the aquatic Lebanese environments,” Environment Pollution, vol. 295, no. 2, pp. 503–511, 2005.
[54]
K. Kümmerer, “Antibiotic in the aquatic environment. A review: part 2,” Chemosphere, vol. 75, pp. 435–441, 2009.
[55]
R. Shrivastava, R. K. Upreti, S. R. Jain, K. N. Prasad, P. K. Seth, and U. C. Chaturvedi, “Suboptimal chlorine treatment of drinking water leads to selection of multidrug-resistant Pseudomonas aeruginosa,” Ecotoxicology and Environmental Safety, vol. 58, no. 2, pp. 277–283, 2004.
[56]
S. N. Anuj, D. M. Whiley, T. J. Kidd et al., “Identification of Pseudomonas aeruginosa by a duplex real-time polymerase chain reaction assay targeting the ecfX and the gyrB genes,” Diagnostic Microbiology and Infectious Disease, vol. 63, no. 2, pp. 127–131, 2009.
[57]
X. Li, M. Mehrotra, S. Ghimire, and L. Adewoye, “β-Lactam resistance and β-lactamases in bacteria of animal origin,” Veterinary Microbiology, vol. 121, no. 3-4, pp. 197–214, 2007.
