Escherichia coli 0157:H7 serotipinin izolasyonunda selektif broth ve agar besiyerleri üzerine bir çalışma

Modified tryptic soy broth, lauryl sulfate tryptose broth, modified EC broth, sorbitol MacConkey agar and Fluorocult E. coli O157 agar were tested for the recovery of 3 strains of Escherichia coliO157:H7 against E. colitype 1 and Citrobacter freundii as common competitive microflora of raw meat products. Two-stepped trials showed that modified EC broth and lauryl sulfate tryptose broth were superior to modified tryptic soy broth as selective enrichment media while sorbitol MacConkey agar was clearly better than Fluorocult E. coli O157 agar as a selective agar medium. Recovery of E. coli O157:H7 was difficult when selective enrichment broth-growing cohabitant microflora were present at more than 100 folds of E. coliO157. One percent E. coli O157:H7 in the cohabitant flora was the minimum level for recovery. This is probably more important than the minimum recovery rate of culture media and/or conventional culture methods when E. coli O157:H7 exists without competitive microorganisms.

A study on selective broths and agar media for the isolation of Escherichia coli 0157:H7 serotype

Modified tryptic soy broth, lauryl sulfate tryptose broth, modified EC broth, sorbitol MacConkey agar and Fluorocult E. coli O157 agar were tested for the recovery of 3 strains of Escherichia coliO157:H7 against E. colitype 1 and Citrobacter freundii as common competitive microflora of raw meat products. Two-stepped trials showed that modified EC broth and lauryl sulfate tryptose broth were superior to modified tryptic soy broth as selective enrichment media while sorbitol MacConkey agar was clearly better than Fluorocult E. coli O157 agar as a selective agar medium. Recovery of E. coli O157:H7 was difficult when selective enrichment broth-growing cohabitant microflora were present at more than 100 folds of E. coliO157. One percent E. coli O157:H7 in the cohabitant flora was the minimum level for recovery. This is probably more important than the minimum recovery rate of culture media and/or conventional culture methods when E. coli O157:H7 exists without competitive microorganisms.

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  • 1. Doyle, P.D., Zhao, T., Meng, J., Zhao, S. Escherichia coli O157:H7. In: M.P. Doyle, L.R. Beuchat, T.J. Montville (Eds.) Food Microbiology: Fundamentals and Frontiers. American Society of Microbiology, Washington D. C. pp. 171-191, 1997.
  • 2. Venkateswaren, K., Kamijoh, Y., Ohashi, E. and Nakanishi, H. A simple filtration technique to detect enterohemorrhagic Escherichia coli O157:H7 and its toxins in beef by multiplex PCR. Appl. Environ. Microbiol. 1997; 63, 4127-4131.
  • 3. Krishnan, C., Fitzgerald, V.A., Dakin, S.J. and Behme, R.J. Laboratory investigation of outbreak of hemorrhagic colitis caused by Escherichia coli O157:H7. J. Clin. Microbiol. 1987; 25, 1043- 1047.
  • 4. Szabo, R.A., Todd, E.C.D. and Jean, A. A method to isolate Escherichia coli O157:H7 from food. J. Food Protect. 1986; 49, 768-772.
  • 5. Anonymous. Singlepath E. coli O157:H7. Leaflet Merck Microbiology W120 133 081 198 E. Merck Co. Darmstad, 1998.
  • 6. Blais, B.W., Booth, B.A., Phillippe, L. and Pandian, S. Polymacron enzyme immunoassay system for detection of Escherichia coli O157:H7 inoculated into foods. J. Food Prot. 1997; 60, 98-101.
  • 7. He, Y., Keen, J.E., Westerman, R.B. and Littledike, E.T. Monoclonal antibodies for detection of the H7 antigen of Escherichia coli. Appl. Environ. Microbiol. 1996; 62, 3325-3332.
  • 8. Karch, H., Mittmann, C.J., Aleksic, S. and Datz, M. Isolation of enterohemorrhagic Escherichia coli O157 strains from patients with hemolytic uremic syndrome by using immunomagnetic separation, DNA based methods and direct culture J. Clin. Microbiol. 1996; 34, 516-519.
  • 9. Doyle, M.P. and Schoeni, J.L. Isolation of Escherichia coli O157:H7 from retail fresh meats and poultry. Appl. Environ. Micr. 1997; 53, 2394-2396.
  • 10. Okrend, A.J.G., Rose, B.E. and Bennett, B.A. A screening method for the isolation of Escherichia coli O157:H7 from ground beef. J. Food Protect. 1990; 53, 249-252.
  • 11. March, S.B. and Ratnam, S.J. Sorbitol MacConkey medium for detection of Escherichia coli O157:H7 associated with hemorrhagic colitis. J. Clin. Microbiol. 1986; 23, 869-872.
  • 12. Chapman, P. A., Siddons C. A., Zad›k P. M. and Jewes, L. An improved selective medium for the isolation of Escherichia coli O157. J. Med. Microbiol. 1991; 35, 107-110.
  • 13. Heuvelink, A.E., Zwqrtkruis-Nahuis, T.M. and Boer, E.D. Evaluation of media and test kits for the detection of Escherichia coli O157 from minced beef. J. Food Protect. 1997; 60, 817- 824.
  • 14. Okrend, A.J.G., Rose, B.E. and Lattuda, C.P. 5-bromo-4-chloro-3- indoxyl-ß-D-glucuronide in sorbitol MacConkey agar to aid in the isolation of Escherichia coli O157: H7 from ground beef. J. Food Protect. 1990; 53, 941-943.
  • 15. Hitchins, A.D., Feng, P., Watkins, W.D., Rippey, S.R., and Chandler, S.A. Escherichia coli and coliform bacteria. In: Bacteriological Analytical Manual BAM/AOAC 8th Edition. AOAC International, Gaithersburg, pp 4.01-4.29, 1998.
  • 16. Okrend, A.J.G., Rose, B.E. and Lattuda, C.P. Use of 5-Bromo-4 Chloro-3-Indoxy-ß-D-Glucuronide in Sorbitol MacConkey Sorbitol Agar to Aid in the Isolation of Escherichia coli O157:H7 from Ground Beef. J Food Prot 1990; 53, 936-940.
  • 17. Polumbo, S.A., Pickard, A. and Call, J.E. Population changes and verotoxin production of enterohemorrhagic Escherichia coli strains inoculated in milk and ground beef held at low temperatures. J Food Protect 1977; 60, 746-750.
  • 18. Halkman, A. K., Noveir, M.R. and Do_an, H.B. Investigation of E. coli O157:H7 in Animal Origin Foods. Scientific and Technical Research Council of Turkey, Veterinary and Husbandry Research Group Project no TUBITAK-VHAG-1192 (in Turkish with English abstract, unpublished), 1998.
  • 19. Anonymous. Bergey’s Manual of Determinative Bacteriology. Eds. J. Holt et al. Williams & Wilkinson Co. Maryland, 787 p, 1994.
  • 20. Anonymous. Merck Microbiology Manual. E. Merck Co. Darmstad, 1994.
  • 21. Kleanthous, H., Fry, N.K., Smith, H.R., Gross, R.J. and Rowe, B. The use of sorbitol-MacConkey agar in conjunction with a specific antiserum for the detection of Vero cytotoxin-producing strains of Escherichia coli O157. Epidem. Inf. 1988; 101, 327-335.
  • 22. Schothorst, M., Renaud,A. and van Beek C. Salmonella isolation using RVS broth and MLCB agar. Food Microbiology 1987; 4, 11- 18.
  • 23. Smith, H.R. and Scotland, S.M. Vero cytotxin-producing strains of Escherichia coli. J. Med Microbiol. 1988; 26, 77-85.
Turkish Journal of Veterinary and Animal Sciences-Cover
  • ISSN: 1300-0128
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK