Shigatoxin producing Escherichia coli O157 and non-O157 serotypes in producer-distributor bulk milk

Abstract

Several recent large outbreaks of gastrointestinal diseases have highlighted the threat posed by morbidity and mortality associated with shigatoxin-producing Escherichia coli (STEC). Furthermore, the treatment of STEC infections is now threatened by the emergence of antibiotic resistance which is an alarming health concern in the world of medicine. The most implicated STEC in foodborne disease outbreaks across the globe is O157 serotype, although some emerging STEC non-O157 serotypes are increasingly becoming recognised as foodborne pathogens of important public health concern. This study was undertaken to characterise bacterial species in raw and pasteurised producer-distributor bulk milk (PDBM) with specific emphasis on E. coli and other Enterobacteriaceae. E. coli was further investigated for the prevalence and distribution of virulence factors (stx 1, stx 2 and hlyA), serotypes and antibiotic resistance patterns, which also included extended-spectrum ?-lactamase (ESBL) producing capacity. Subsequently, the study further estimated the haemolytic uraemic syndrome (HUS) risk associated with the consumption of STEC contaminated PDBM and also estimated the resulting burden of illness that may be associated with the consumption of such milk in South Africa (SA). A total of 258 PDBM samples were collected, using convenience sampling, from outlets (purchase points) in eight different geographical provinces in SA. Isolation, detection and enumeration of total aerobic bacteria, coliforms and E. coli were carried out using 3M E. coli /coliform petrifilm plates. Matrix assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) was used for the rapid identification of the bacterial isolates. The identification of E. coli was confirmed using PCR of the uidA gene. Further characterisation of E. coli into serogroups, identification of virulence factors and antibiotic resistance profiles were then performed. E. coli O157 was characterised using selective media and confirmed using mismatch amplification mutation assay (MAMA)-multiplex PCR. Identification of E. coli -serogroups was carried out by the restriction of amplified O-antigen gene cluster (rfb-RFLP), coupled with serum agglutination assay. Virulence factors (stx 1, stx 2 and hlyA) were determined using both phenotypic and genotypic characterisation. Antimicrobial agent susceptibility tests and detection of extended spectrum beta-lactamase (ESBL) producing capacity of the E. coli isolates were performed using phenotypic characterisation. Finally, a quantitative risk assessment for STEC in PDBM was also conducted. More than 60% of the PDBM samples were found not to be fit for human consumption on the basis of the minimum standards prescribed in the Foodstuffs, Cosmetics and Disinfectants Act (Act 54 of 1972). Raw and pasteurised PDBM was contaminated with a wide diversity of Enterobacteriaceae species, which included spoilage microbiota.