Biolog identification of non-sorbitol fermenting bacteria isolated on E. coli O157 selective CT-SMAC agar

Abstract

E. coli O157:H7 is recognised as an important human pathogen world-wide and has been associated with diseases such as haemorrhagic colitis (HC), haemolytic uraemic syndrome (HUS) and thrombotic thrombocytopaenic purpura (TTP). Accurate laboratory detection of E. coli O157:H7 is important for diagnostic purposes and to justify epidemiological data on E. coli O157:H7. A well-known phenotypic characteristic of E. coli O157:H7 bacteria is their inability to ferment sorbitol. This characteristic is often used to isolate these organisms from food and water using selective agar medium such as SMAC. However, the high number of false positive results obtained by a number of researchers when selectively screening for E. coli O157: H7 on CT-SMAC has prompted an investigation to determine which other sorbitol-negative bacteria also grow on CT-SMAC. The agar medium used for the investigation consisted of Sorbitol MacConkey agar (SMAC) supplemented with Cefiximetellurite (CT). All sorbitol-negative colonies obtained from CT-SMAC, after selective enrichment and IMS were identified using the Biolog microbial identification system. The majority of sorbitol-negative isolates identified were Burkholderia, Pseudomonas, Vibrio and Aeromonas spp. Only two E. coli O157:H7 isolates were identified with Biolog and confirmed with a polymerase chain reaction (PCR) specific for the shiga toxin 1 (Stx1) genes and with O157 and H7 antisera. The inability of the CT-SMAC agar medium to specifically select for E.coli O157:H7 was confirmed by the results of this study. These observations call for further improvement of affordable methods for the selective isolation of E. coli O157:H7 in the presence of large numbers of interfering bacteria capable of growing on CT-SMAC.