The detection of Shiga toxin-producing Escherichia coli in diagnostic bovine faecal samples using vancomycin-cefixime-cefsulodin blood agar and PCR.
The prevalence of complex Shiga toxin-producing Escherichia coli (STEC), i.e. STEC containing accessory virulence factors intimin (eaeA) and/or enterohaemorrhagic E. coli haemolysin (ehxA) and their serotypes were determined in diagnostic bovine faecal samples processed during a 3 months period. The presence of complex STEC was determined using PCR and vancomycin-cefixime-cefsulodin blood agar (BVCCA) using a dual approach which involved (i) direct culture of faecal samples on BVCCA followed by mutiplex PCR of BVCCA positive colonies and (ii) culture of faecal samples enriched in modified EC (mEC) broth (with a complex STEC profile determined by PCR) on BVCCA followed by multiplex PCR of BVCCA positive colonies. Using both techniques complex STEC were isolated from 23 (18.7%) of the 123 faecal samples. Complex STEC were isolated from 14 faecal samples by direct culture on BVCCA and 13 faecal samples yielded complex STEC by culture of mEC broths with a complex STEC profile on BVCCA. Only four samples were positive using both techniques. The serotypes isolated included O5:H-, O26:H-, O26:H11, O91:H21, O111:H-, O111:H8, O104:H11, O113:H21 and O157:H8. This study confirms that non-O157 STEC can be isolated from bovine faeces and that they carry types associated with human disease. This work also demonstrates that the use of a dual approach is advisable to increase the likelihood of isolating complex STEC.
Hornitzky
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Elizabeth Macarthur Agricultural Institute, New South Wales Agriculture, National Escherichia coli Reference, Private Mail Bag, 8, Camden, NSW 2570 Australia. michael.hornitzky@agric.nsw.gov.au
Bettelheim
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Djordjevic
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D018829:Adhesins, Bacterial; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000235:genetics; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D001431:Bacteriological Techniques; D002352:Carrier Proteins; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D020682:Cefixime; D002441:Cefsulodin; D003470:Culture Media; D003967:Diarrhea / Q000382:microbiology / Q000662:veterinary*; D004926:Escherichia coli / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism / Q000472:pathogenicity; D029968:Escherichia coli Proteins; D005243:Feces / Q000382:microbiology*; D006460:Hemolysin Proteins / Q000032:analysis / Q000235:genetics; D016133:Polymerase Chain Reaction; D012703:Serotyping; D022622:Shiga Toxin 1 / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D022641:Shiga Toxin 2 / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D014640:Vancomycin; D014774:Virulence
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D003470:Culture Media; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; C073819:eaeA protein, E coli; D014640:Vancomycin; D020682:Cefixime; D002441:Cefsulodin