Comparison of four methods for isolation of Yersinia enterocolitica from raw and pasteurized milk from northern Iran

Abstract

Four methods for isolation of Yersinia enterocolitica from raw and pasteurized milk from northern Iran were compared. Three hundred and ten raw milk samples were collected from tanks on their arrival at various central dairies, and 40 pasteurized milk samples were collected from tanks on their arrival at a manufacturing plant. Each sample was examined for the presence of Y. enterocolitica by (1) direct culture; (2) enrichment in double-strength buffered peptone water at 4 degrees C for 1 month; (3) enrichment in modified Rappaport medium at room temperature for 72 h after a preenrichment in double-strength peptone water at 4 degrees C for 1 month; and (4) enrichment in a medium containing sucrose, tris (hydroxymethyl) aminomethane, sodium azide, and ampicillin at 28 degrees C for 48 h after a preenrichment in double-strength peptone water at 4 degrees C for 1 month. All samples and enrichments were spread on MacConkey agar plus calcium chloride and Tween 80, Yersinia selective agar, and Hektoen medium plus ampicillin. Five samples (1.6%) of raw milk but no pasteurized milk samples were positive for Y. enterocolitica. No Y. enterocolitica were recovered by methods 1 or 2. Y. enterocolitica were recovered from 2 samples by method 3 followed by culture on Yersinia selective agar, and from 5 samples by method 4 followed by culture on Hektoen medium plus ampicillin. The isolates were biotype 1A or 1B, serotype O:7-13 or O:9 and phage type Xo or Xz. All isolates were resistant to ampicillin and amoxicillin, and sensitive to tetracycline, streptomycin, chloramphenicol, and trimethoprim-sulfamethoxazole.