Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Comparison of Biochemical Identification to Detect Pathogenic Escherichia coli in Fresh Vegetables

신선편이 엽채류의 병원성 E. coli 검출을 위한 생화학적동정법 비교 분석

  • Choi, Yukyung (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Lee, Heeyoung (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Lee, Soomin (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Kim, Sejeong (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Ha, Jimyeng (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Lee, Jeeyeon (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Oh, Hyemin (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Yoon, Yohan (Department of Food and Nutrition, Sookmyung Women's University)
  • 최유경 (숙명여자대학교 식품영양학과) ;
  • 이희영 (숙명여자대학교 식품영양학과) ;
  • 이수민 (숙명여자대학교 식품영양학과) ;
  • 김세정 (숙명여자대학교 식품영양학과) ;
  • 하지명 (숙명여자대학교 식품영양학과) ;
  • 이지연 (숙명여자대학교 식품영양학과) ;
  • 오혜민 (숙명여자대학교 식품영양학과) ;
  • 윤요한 (숙명여자대학교 식품영양학과)
  • Received : 2016.10.03
  • Accepted : 2016.10.30
  • Published : 2016.12.30
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Abstract

The objective of this study was to isolate pathogenic Escherichia coli from fresh vegetables with selective media and Petrifilm, and identify a suspicious colony using biochemical identification. Twenty gram of lettuce, twenty gram of cabbage and ten gram of sprout were prepared, and a 5-strain mixture of pathogenic E. coli (Enterohemorrhagic E. coli NCCP11142, Enterotoxigenic E. coli NCCP14037, Enteropathogenic E. coli NCCP14038, Enteroaggregative E. coli NCCP14039, Enteropathogenic E. coli NCCP15661) was inoculated to obtain 1, 2 and 3 log CFU/g. Eighty to ninety milliliter of buffered peptone water (BPW) was placed and pummeled for 60 s. As a results, the Petrifilm method was all positive, but enrichment method of qualitative analysis was negative except for 3-log CFU/g inoculated lettuce. Regarding biochemical identification of pathogenic E. coli, the identification rates were dependent on type of methods and vegetables; lettuce: API 20E 100% (44/44), Microgen GNA 100% (44/44) and Food System 66.7% (10/15), cabbage: API 20E 64.7% (22/34), Microgen GNA 50% (16/32) and Food System 60% (9/15), sprout: API 20E 65.1% (28/43), Microgen GNA 62.3% (27/43) and Food System 53.3% (8/15). These results could be useful in determining an appropriate method to detect pathogenic E. coli in fresh vegetables.

본 연구에서는 선택배지와 건조필름을 이용하여 신선편이 엽채류 3종(양상추, 양배추, 어린잎채소)에서 병원성 E. coli를 분리하였고, 의심집락 동정을 위해 생화학적 분석법을 사용하여 동정한 후 결과를 비교 분석하였다. 양상추 20 g, 양배추 20 g, 어린잎채소 10 g에 병원성 E. coli 혼합 균액(Enterohemorrhagic E. coli NCCP11142, Enterotoxigenic E. coli NCCP14037, Enteropathogenic E. coli NCCP14038, Enteroaggregative E. coli NCCP14039, Enteropathogenic E. coli NCCP15661)을 최종농도가 1, 2, 3 log CFU/g이 되도록 접종하였고, BPW (80~90 ml)을 넣은 후 60초 동안 균질화하여 분석하였다. 연구결과, 모든 시료에서 건조필름 시험 양성, 양상추 시료 일부(최종농도 3 log CFU/g 접종시료)를 제외한 모든 시료에서 증균배양법을 이용한 정성시험결과가 음성으로 나타났다. 증균배양과 건조필름 시험을 통해 분리한 병원성 E. coli를 이용하여 생화학적 분석을 실시한 결과, 양상추에서 분리한 병원성 E. coli의 경우, API 20E 100% (44/44), Microgen GNA 100% (44/44), Food System 66.7% (10/15)의 동정률이 나타났다. 양배추의 경우, API 20E 64.7% (22/34), Microgen GNA 50% (16/32), Food System 60% (9/15), 어린잎채소의 경우, API 20E 65.1% (28/43), Microgen GNA 62.3% (27/43), Food System 53.3% (8/15)가 병원성 E. coli로 동정되었다. 본 연구의 결과는 신선편이 엽채류에 대한 병원성 E. coli 검출법 선택에 유용하게 이용될 것으로 판단된다.

Keywords

References

  1. National Agricultural Products Quality Management Service (http://www.naqs.go.kr).
  2. Jo, M.J., Jeong, A.R., Kim, H.J., Lee, N., Oh, S.W., Kim, Y.J., Chun, H.S., Koo, M.: Microbiological quality of freshcut produce and organic vegetables. Korean J. Food Sci. Technol., 43, 91-97 (2011). https://doi.org/10.9721/KJFST.2011.43.1.091
  3. National Agricultural Products Quality Management Service. Available at: http://www.naqs.go.kr/multiboard/board/detail.naqs?menu=MN30511&groupno=283&cate=0&page=1&seq=48438&schType=&schKeyword= Accessed on September 28, 2016 (2016).
  4. Kang, T.M., Cho, S.K., Park, J.Y., Song, K.B., Chung, M.S., Park, J.H.: Analysis of microbial contamination of sprouts and fresh-cut salads in a market. Korean J. Food Sci. Technol., 43, 490-494 (2011). https://doi.org/10.9721/KJFST.2011.43.4.490
  5. Choi, J.W., Park, S.Y., Yeon, J.H., Lee, M.J., Chung, D.H., Lee, K.H., Kim, M.G., Lee, D.H., Kim, K.S., Ha, S.D.: Microbial contamination levels of fresh vegetables distributed in markets. J. Food Hyg. Saf., 20, 43-47 (2005).
  6. FDA: Guidance for Industry: Guide to Minimize Microbial Food Safety Hazards of Leafy Greens. Available at: http://www.fda.gov/Food/GuidanceRegulation/GuidanceDocumentsRegulatoryInformation/ucm174200.htm. Accessed on September 28, 2016 (2009).
  7. EFSA (European Food Safety Authority): Shiga toxin-producing E. coli (STEC) O104:H4 2011 outbreaks in Europe: Taking Stock, EFSA Journal, 9, 2390 (2011). https://doi.org/10.2903/j.efsa.2011.2390
  8. Lee, C., Sung, D., Oh, S.: Profiling and priority selection of foodborne pathogens in fresh produce. J. Food Hyg. Saf., 27, 356-365 (2012). https://doi.org/10.13103/JFHS.2012.27.4.356
  9. Kim, S.R., Choi, S.Y., Seo, M.K., Lee, J.Y., Kim, W.I., Yoon, Y., Ryu, K.Y., Yun, J.C., Kim, B.S.: Establishment of sample preparation method to enhance recovery of food-borne pathogens from produce. J. Food Hyg. Saf., 28, 279-285 (2013). https://doi.org/10.13103/JFHS.2013.28.3.279
  10. Food and Agriculture Organization/World Health Organization: Microbiological hazards in fresh leafy vegetables and herbs: microbiological risk assessment series 14 meeting report. (World Health Organization. eds.) FAO/WHO, pp. 9 (2008).
  11. Wang, G., Clark, G.C., Rodgers, G.F.: Detection in Escherichia coli of the genes encoding the major virulence factors, the genes defining the O157:H7 serotype, and components of the type 2 shiga toxin family by multiplex PCR. J. Clin. Miccrobiol., 40, 3613-3619 (2002). https://doi.org/10.1128/JCM.40.10.3613-3619.2002
  12. Mazaheri, S., Salmanzadeh-Ahrabi, S., Falsafi, T., Aslani, M.M.: Isolation of Enteropathogenic Escherichia coli from lettuce samples in Tehran. Gastroenterol. Hepatol. Bed Bench, 7, 38-42 (2014).
  13. Kim, S.R., Lee, J.Y., Lee, S.H., Kim, W.I., Park, K.H., Yun, H.J., Kim, B.S., Chung, D.H., Yun, J.C., Ryu, K.Y.: Evaluation of microbiological safety of lettuce and cultivation area. J. Food Hyg. Saf., 26, 289-295 (2011).
  14. Sadek, Z.I., Hosny, I.M., El-Kholy, W.I., El-Dairouty, R.K.: Comparative investigations for detection of foodborne microorganisms in Egyptian hard cheese "Ras" using conventional and fast biochemical tests, Glob. Vet., 3, 189-195 (2009).
  15. Saeed, A.Y., Mazin, H., Saadi, A., Hussein, S.O.: Detection of Escherichia coli O157 in vegetables. J. Agric. Vet. Sci., 6, 16-18 (2013).
  16. Shearer, A.E.,, Strapp, C.M., Joerger, R.D.: Evaluation of a polymerase chain reaction-based system for detection of Salmonella enteritidis, Escherichia coli O157:H7, Listeria spp., and Listeria monocytogenes on fresh fruits and vegetables. J. Food Prot., 64, 788-795 (2001). https://doi.org/10.4315/0362-028X-64.6.788
  17. Jung, K.S., Roh, E.J., Ryu, K.Y., Kim, W.I., Park, K.H., Lee, D.H., Kim, K.H., Yun, J.C., Heu, S.G.: Monitoring of pathogenic bacteria in organic vegetables from korean market. Korean J. Soil Sci. Fert., 45, 560-564 (2012). https://doi.org/10.7745/KJSSF.2012.45.4.560
  18. Barry, A.L., Badal, R.E.: Rapid identification of Enterobacteriaceae with the Micro-ID system versus API 20E and conventional media. J. Clin. Microbiol., 10, 293-298 (1979).