Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
권호 표지

Multiple Confirmation and RAPD-genotyping of Enterobacter sakazakii Isolated from Sunsik

선식에서 분리한 Enterobacter sakazakii의 복합동정 및 RAPD를 이용한 genotyping

  • Choi, Jae-Won (Food Safety Research Division, Korea Food Research Institute) ;
  • Kim, Yun-Ji (Food Safety Research Division, Korea Food Research Institute) ;
  • Lee, Jong-Kyung (Food Safety Research Division, Korea Food Research Institute) ;
  • Kim, Young-Ho (Food Safety Research Division, Korea Food Research Institute) ;
  • Kwon, Ki-Sung (Center for Food Evaluation, Korea Food and Drug Adminstration) ;
  • Hwang, In-Gyun (Center for Food Evaluation, Korea Food and Drug Adminstration) ;
  • Oh, Se-Wook (Food Safety Research Division, Korea Food Research Institute)
  • 최재원 (한국식품연구원 안전성연구단) ;
  • 김윤지 (한국식품연구원 안전성연구단) ;
  • 이종경 (한국식품연구원 안전성연구단) ;
  • 김영호 (한국식품연구원 안전성연구단) ;
  • 권기성 (식품의약품안전청 식품평가부) ;
  • 황인균 (식품의약품안전청 식품평가부) ;
  • 오세욱 (한국식품연구원 안전성연구단)
  • Published : 2008.02.28
Download PDF
⟨ Previous Next ⟩

Abstract

Enterobacter sakazakii is implicated in severe forms of neonatal infections such as meningitis and sepsis. This organism has been isolated from a wide range of foods, including cheese, vegetables, grains, herbs, and spices, but its primary environment is still unknown. Generally, dried infant milk formula has been epidemiologically identified as the source of E. sakazakii. Sunsik (a powdered mixture of roasted grains and other foodstuffs) is widely consumed in Korea as a side dish or energy supplement. Sunsik is consumed without heat treatment; thus, lacking an additional opportunity to inactivate foodborne pathogens. Therefore, its microbiological safety should be guaranteed. In this study, the prevalence of E. sakazakii was monitored in 23 different sunsik component flours, using FDA recommended methods; but E. sakazakii medium (Neogen) and Chromogenic E. sakazakii medium (Oxoid) were used as the selective media. In total, presumptive E. sakazakii strains were isolated from 8 different sunsik powders. Subsequently, an API 20E test was conducted, and 15 strains from 5 different sunsik flours (sea tangle, brown rice, non-glutinous rice, cheonggukjang, dried anchovy) were confirmed as E. sakazakii. Fifteen strains were again confirmed by PCR amplification, using three different primer sets (tDNA sequence, ITS sequence, 16S rRNA sequence), and compared to ATCC strains (12868, 29004, 29544, 51329). They were once again confirmed by their enzyme production profiles using an API ZYM kit. Finally, RAPD (random amplified polymorphic DNA)-genotyping was carried out as a monitoring tool to determine the contamination route of E. sakazakii during processing.

시판되고 있는 선식 원료를 수거하여 최근 새로운 식중독균으로 보고되고 있는 Enterobacter sakazakii 분리 실험을 실시하였다. 그 결과 총 23종의 선식 원료 중 8개의 선식 원료에서 E.sakazakii로 추정되는 콜로니를 분리할 수 있었으며 API 20E kit를 이용하여 1차적으로 동정한 결과, 다시마 분말, 멸치 분말, 현미 분말, 청국장 분말 및 멥쌀 분말에서 E. sakazakii를 분리할 수 있었다. 이후 3 종의 primer를 이용한 PCR을 실시하여 2차적으로 동정하였다. 또한, 분리된 균주에 대한 RAPD-PCR을 실시하여 최종적으로 8종의 분리균으로 molecular typing을 할 수 있었다.

Keywords

References

  1. Farmer JJ, Asbury MA, Hickman FW, Brenner DJ. The Enterobacteriaceae study group (USA). Enterobacter sakazakii: A new species of 'Enterobacteriaceae' isolated from clinical specimens. Int. J. Syst. Bacteriol. 30: 569-584 (1980) https://doi.org/10.1099/00207713-30-3-569
  2. Iverson C, Forsythe SJ. Risk profiles of Enterobacter sakazakii, an emergent pathogen associated with infant milk formula. Trends Food Sci. Tech. 14: 443-454 (2003) https://doi.org/10.1016/S0924-2244(03)00155-9
  3. Dennison SK, Morris J. Multiresistant Enterobacter sakazakii wound infection in an adult. Infect. Med. 19: 533-535 (2002)
  4. Emery CL, Weymouth LA. Detection and clinical significance of extended-spectrum beta-lactamases in a tertiary-care medical center. J. Clin. Microbiol. 35: 2061-2067 (1997)
  5. Hawkins RE, Lissner CR, Sanford JP. Enterobacter sakazakii bacteremia in an adult. South. Med. J. 84: 793-795 (1991) https://doi.org/10.1097/00007611-199106000-00033
  6. Jimenez E, Gimenez C. Septic shock due to Enterobacter sakazakii. Clin. Microbiol. Newslett. 4: 30 (1982) https://doi.org/10.1016/S0196-4399(82)80129-3
  7. Pribyl C, Salzer R, Beskin J, Haddad RJ, Pollock B, Beville R, Holmes B, Mogabgab WJ. Aztreonam in the treatment of serious orthopedic infections. Am. J. Med. 78: 51-56 (1985) https://doi.org/10.1016/0002-9343(85)90205-0
  8. Lai KK. Enterobacter sakazakii infections among neonates, infants, children, and adults. Case reports and a review of the literature. Medicine 80: 113-122 (2001) https://doi.org/10.1097/00005792-200103000-00004
  9. Bar-Oz B, Preminger A, Peleg O, Block C, Arad I. Enterobacter sakazakii infection in the newborn. Acta Paediatr. 90: 356-358 (2001) https://doi.org/10.1111/j.1651-2227.2001.tb00319.x
  10. Biering G, Karlsson S, Clark NC, Jonsdottir KE, Ludvigsson P, Steingrimsson O. Three cases of neonatal meningitis caused by Enterobacter sakazakii in powdered milk. J. Clin. Microbiol. 27: 2054-2056 (1989)
  11. Clark NC, Hill BC, O'Hara CM, Steingrimsson O, Cooksey RC. Epidemiologic typing of Enterobacter sakazakii in two neonatal nosocomial outbreaks. Diagn. Micr. Infec. Dis. 13: 467-72 (1990) https://doi.org/10.1016/0732-8893(90)90078-A
  12. Muytjens HL, Kollee LA. Enterobacter sakazakii meningitis in neonates: Causative role of formula? Pediatr. Infect. Dis. J. 9: 372-373 (1990) https://doi.org/10.1097/00006454-199005000-00016
  13. Muytjens HL, Zanen HC, Sonderkamp HJ, Kollee LA, Wachsmuth IK, Farmer JJ. Analysis of eight cases of neonatal meningitis and sepsis due to Enterobacter sakazakii. J. Clin. Microbiol. 18: 115-120 (1983)
  14. Noriega FR, Kotloff KL, Martin MA, Schwalbe RS. Nosocomial bacteremia caused by Enterobacter Sakazakii and Leuconostoc mesenteroides resulting from extrinsic contamination of infant formula. Pediatr. Infect. Dis. J. 9: 447-449 (1990) https://doi.org/10.1097/00006454-199006000-00018
  15. Simmons BP, Gelfand MS, Haas M, Metts L, Ferguson F. Enterobacter sakazakii infections in neonates associated with intrinsic contamination of a powdered infant formula. Infect. Cont. Hosp. Ep. 10: 398-401 (1989) https://doi.org/10.1086/646060
  16. Van Acker J, De Smet F, Muyldermans G, Bougatef A, Naessens A, Lauwers S. Outbreak of necrotizing enterocolitis associated with Enterobacter sakazakii in powdered milk formula. J. Clin. Microbiol. 39: 293-297 (2001) https://doi.org/10.1128/JCM.39.1.293-297.2001
  17. Muytjens HL, Roelofs-Willemse H, Jaspar GH. Quality of powdered substitutes for breast milk with regard to members of the family Enterobacteriaceae. J. Clin. Microbiol. 26: 743-746 (1988)
  18. Nazarowec-White M, Farber JM. Incidence, survival, and growth of Enterobacter sakazakii in infant formula. J. Food Protect. 60: 226-230 (1997) https://doi.org/10.4315/0362-028X-60.3.226
  19. Wyeth Nutrition. Wyeth Nutrition initiates voluntary recall of selected store brand infant formula from U.S. marketplace. http:// www.wyeth.com/news?nav=display&navTo=/wyeth_html/home/ news/pressreleases/2002/1146452500787.html. Accessed Nov. 1, 2002
  20. The Food Safety Authority of Ireland. Pregestimil infant formula recall. http://www.fsai.ie/alerts/archive/fa20050120.asp. Accessed Jan. 20, 2005
  21. Gurtler JB, Kornacki JL, Beuchat LR. Enterobacter sakazakii: A coliform of increased concern to infant health. Int. J. Food Microbiol. 104: 1-34 (2005) https://doi.org/10.1016/j.ijfoodmicro.2005.02.013
  22. Kandhai MC, Reij MW, Gorris LG, Guillaume-Gentil O, van Schothorst M. Occurrence of Enterobacter sakazakii in food production environments and households. Lancet 363: 39-40 (2004) https://doi.org/10.1016/S0140-6736(03)15169-0
  23. Enterobacter sakazakii and other microorganisms in powdered infant formula: meeting report, MRA Series 6. Microbiological Risk Assessment Series, No. 6 ISBN: 92 4 156262 5 (WHO) http://www.who.int/foodsafety/publications/micro/es.pdf. Accessed Dec. 30, 2006
  24. Chung SS, Han YS. Consumer's recognition, nutrient composition, and safety evaluation of commercial sunsik and saengsik. Korean J. Food Culture 18: 235-243 (2003)
  25. FDA. Food and Drug Administration, Isolation and enumeration of Enterobacter sakazakii from dehydrated powdered infant formula. http://www.cfsan.fda. gov/~comm/mmesakaz.html. Accessed Sep. 19, 2003
  26. Iverson C, Druggan P, Forsythe S. A selective differential medium for Enterobacter sakazakii, a preliminary study. Int. J. Food Microbiol. 96: 133-139 (2004) https://doi.org/10.1016/j.ijfoodmicro.2004.01.024
  27. Oh SW, Kang DH. Fluorogenic selective and differential medium for isolation of Enterobacter sakazakii. Appl. Environ. Microb. 70: 5692-5694 (2004) https://doi.org/10.1128/AEM.70.9.5692-5694.2004
  28. FDA. http://www.cfsan.fda.gov/~ebam/bam-a2.html#excl. Accessed Dec. 29, 2006
  29. Clementino MM, Filippis ID, Nascimento CR, Branquinho R, Rocha CL, Martins OB. PCR analyses of tRNA intergenic spacer, 16S-23S internal transcribed spacer, and randomly amplified polymorphic DNA reveal inter- and intraspecific relationship of Enterobacter cloacae strains. J. Clin. Microbiol. 39: 3865-3870 (2001) https://doi.org/10.1128/JCM.39.11.3865-3870.2001
  30. Keyser M, Withuhn RC, Ronquest LC, Britz T. Treatment of winery effluent with upflow anaerobic sludge blanket (UASB)- granular sludges enriched with Enterobacter sakazakii. Biotechnol. Lett. 25: 1893-1898 (2003) https://doi.org/10.1023/B:BILE.0000003978.72266.96
  31. Lehner A, Tasara T, Stephan R. 16S rRNA genes based analysis of Enterobacter sakazakii strains from different sources and development of a PCR assay for identification. BMC Microbiol. 4: 43-50 (2004) https://doi.org/10.1186/1471-2180-4-43