Analysis of Waterborne Pathogenic Bacteria among Total Coliform Positive Samples in the Groundwater of Chungcheongnam-do Province, Korea

충남지역 지하수에서 분리한 총대장균군 양성시료 중 수인성 병원균의 분석

Yu, Jungho;Wang, Changkeun;Shin, Inchul;Kim, Donguk;Park, Kwisung

  • Received : 2016.03.09
  • Accepted : 2016.05.27
  • Published : 2016.06.30


Objectives: To ensure the microbiological safety of groundwater, it was confirmed whether waterborne pathogenic bacteria in groundwater samples tested positive for total coliforms in the Chungcheongnam-do Province region. Methods: Total colony counts, total coliforms and fecal coliforms were tested according to the process mandated by the drinking water quality testing standards of Korea. DNA was extracted from the samples, tested positive for total coliforms, and then subjected to real-time PCR to detect waterborne pathogenic bacteria. Results: A total of 115 samples were inadequate for drinking water. Thirty-one cases (27%) showed positive for fecal coliforms and nine cases (7.8%) showed total colony counts exceeding drinking water standards. Twenty-seven cases (23.5%) showed three items (total colony counts, total coliforms and fecal coliforms). Using the real-time PCR method, waterborne pathogens were detected in 57 cases (49.6%) in 115 samples. Seventy-eight cases of waterborne pathogenic bacteria were detected (including duplications): 27 cases of pathogenic E. coli (EPEC (19), ETEC (5), EHEC (1), EAEC (1) and EIEC (1)); 45 of Bacillus cereus; two of Yersinia spp.; two of Salmonella spp.; one of Staphylococcus aureus; one of Clostridium perfringens. Conclusion: The real-time PCR method can offer rapid and accurate detection of waterborne pathogenic bacteria. Therefore, this assay could be an alternative to conventional culture methods and can further ensure the microbiological safety of groundwater.


Groundwater;real-time PCR;total coliforms;waterborne pathogenic bacteria


  1. Park HK, Jung EY, Jung JM, Yu PJ. Detection and distribution of bacterial pathogens in raw water and during water treatment process by polymerase chain reaction. J life sci. 2007; 17(10): 1374-1380.
  2. Lee IH, Kim SK, Choi YH, Kim JS. Distribution and Characteristics of Coliform Bacteria in Groundwater of Yeungnam Province. Korean J Microbiol. 2006; 42(2): 95-102.
  3. Ministry of environment. Drinking Water Quality Official Test Standards; 2010
  4. Lim HS. Contributing Factors of Infectious Waterborne and Foodborne Outbreaks in Korea. J Korean. Med. Assoc. 2007; 50(7): 582-591.
  5. Ministry of environment. 2013 Statistics of Waterworks. 2014. p.5-6.
  6. Ministry of Land, Infrastructure and Transport (MOLIT), K-water. 2014 Groundwater annual report. 2014. p5.
  7. Ministry of environment. National Institute of Environmental Research. Operating result of Groundwater quality monitoring network in 2013. 2014. p5.
  8. Kim KH, Kim KR, Kim HS, Lee GT, Lee KH. Assessment of Soil and Groundwater Contamination at Two Animal Carcass Disposal Sites. Korean J Soil Sci Fert. 2010; 43: 384-389.
  9. Choi NC, Choi EJ, Kim BJ, Kim SB, Park JA, Park CY. Characterization of Water Quality and the Aerobic Bacterial Population in Leachate Derived from Animal Carcass Disposal. J Eng Geol. 2013; 23(1): 37-46.
  10. Lee JK, Park IH, Yoon KS, Kim HJ, Cho JI, Lee SH, et al. An analysis of epidemiological investigation reports regarding to pathogenic E. coil outbreaks in Korea from 2009 to 2010. J. Fd Hug. Safety. 2012; 27(4): 366-374.
  11. Granum PE. Bacillus cereus and its toxins. J Appl Bacteriol Sym. 1994; 76: 61-66.
  12. Ryu JH. Beuchat LR. Biofilm formation and sporulation by Bacillus cereus on a stainless steel surface and subsequent resistance of vegetative cells and spores to chlorine, chlorine dioxide, and a peroxyacetic acid-based sanitizer. J Food Prot. 2005; 68: 2614-2622.
  13. Choi KB, Lim HS, Lee K, Ha GY, Jung KH, Sohn CK. Epidemiological Investigation for Outbreak of Food Poisoning Caused by Bacillus cereus Among the Workers at a Local Company in 2010. J Prev Med Pub Heal. 2011; 44(2): 65-73.
  14. Stenfors Arnesen LP, Fagerlund A, Granum PE. From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol Rev. 2008; 32(4): 579-606.
  15. Fricker EJ, Fricker CR. Application of the polymerase chain reaction to the identification of Escherichia coli and coliforms in water. Letters Appl. Microbiol. 1994; 57: 320-323.
  16. Seo RJ, Park SH, Lee GH. Microbiological water quality of water purifiers at elementary schools in gunsan area. Korean J Microbiol. 2009; 45(1): 74-81.