No abstract

Shiga toxin-producing Escherichia coli (STEC) O157:H7/NM and some non-O157 STEC are foodborne pathogens. In response to pork-associated O157 STEC outbreaks in Canada, we investigated the occurrence of STEC in Canadian retail raw ground pork during the period of November 1, 2014 and March 31, 2016. Isolated STEC were characterized to determine the Shiga-toxin gene ( stx ) subtype and the presence of virulence genes encoding intimin ( eae ), and enterohemorrhagic E. coli hemolysin (hlyA) . O157 STEC and non-O157 STEC were isolated from 0.11% (1/879) and 2.24% (13/580) of the pork samples. STEC virulence gene profiles containing both eae and hlyA were found only in the O157 STEC ( stx 2a , eae , hlyA ) isolate. The eae gene was absent from all non-O157 STEC isolates. Of the 13 non-O157 STEC isolates, two virulence genes of stx 1a and hlyA were found in four (30.8%) O91:H14 STEC isolates, while one virulence gene of stx 2e, stx 1a , and stx 2a was identified in five (38.5%), two (15.4%) and one (7.7%) STEC isolates respectively of various serotypes. The remaining non-O157 STEC isolate carried stx 2 , but the subtype is unknown as this isolate could not be recovered for sequencing. O91:H14 STEC ( stx 1a, hlyA ) was previously reported in association with diarrhea illnesses, while the other non-O157 STEC isolates identified in this study are not known to be associated with severe human illnesses. Virulence gene profiles identified in this study indicate that the occurrence of non-O157 STEC capable of causing severe human illness is rare in Canadian retail pork. However, O157 STEC in ground pork can occasionally occur, therefore education regarding the potential risks associated with STEC contamination of pork would be beneficial for the public and those in the food industry in order to help reduce foodborne illnesses.

Following two O121 STEC outbreaks linked to wheat flour, this study was conducted to gain baseline information on the occurrence of bacterial pathogens and levels of indicator organisms in wheat flour in Canada. A total of 347 pre-packaged wheat flour samples were analyzed for Salmonella spp., Shiga Toxin-Producing Escherichia coli (STEC), Listeria monocytogenes ( L. monocytogenes ) , aerobic colony count (ACC), total coliforms, and generic Escherichia coli ( E. coli ) . Salmonella spp. and O157 STEC were not detected in any of the samples. L. monocytogenes was identified in two samples (0.6%) at levels below the limit of detection (<0.7 log CFU/g). Non-O157 STEC were isolated from six samples (1.7%) and were characterized for the presence of STEC virulence genes: stx 1, stx 2 and subtypes, eae , hlyA, and aggR . One O103:H25 STEC isolate carried virulence genes ( stx 1 a + eae ) that are known to be capable of causing diarrhea and/or bloody diarrhea in humans. Of the five remaining non-O157 STEC isolates, four carried single stx 2a or stx 2c genes and were considered to have the potential of causing diarrhea. The remaining non-O157 STEC isolate ( stx 2 ), while not a priority non-O157 STEC was not available for sequencing and thus its potential to cause illness is unknown. ACC, total coliforms, and generic E. coli were detected   in 98.8%, 72.6% and 0.6% of the flour samples. The mean counts of ACC were greater in whole-wheat flour as compared to the other flour types tested ( p <0.001). The results of this study suggest that the occurrence of O157 STEC and Salmonella  is low, but the occurrence of non-O157 STEC in wheat flour with the potential to cause human illness of diarrhea is relatively common. Therefore, the consumption of raw flour could increase the likelihood of STEC infections. Further research is merited for potential risk mitigation strategies within the food production system and with consumers.

A preliminary investigation showed that polyoxyethylene emulsifiers contain substantial amounts of "free" polyethylene glycol. An improved method for determining these emulsifiers in foods is presented, in which the emulsifier is extracted with chloroform, "cleaned up" on an alumina column and analysed by thin-layer chromatography with a modified Dragendorff reagent to spray the chromatogram. The method is a t least f 1 5 per cent. accurate down to an emulsifier level of 0.01 per cent. in fats and 0.001 per cent. in baked foods and food mixes. The detection of polyoxyethylene emulsifiers can be carried out a t even lower levels.

Single-strand conformation polymorphism (SSCP) analysis of amplicons produced from a mitochondrial DNA region between the tRNA(Lys) and ATPase8 genes was applied for the detection of animal product within livestock feeds. Identification of prohibited animal (cattle, elk, sheep, deer, and goat) and nonprohibited animal (pig and horse) products from North America was possible based on the differential display of the single-stranded DNA fragments for the different animal species on SSCP gels. This method allowed specific detection and identification of mixed genomic DNA from different animal species. Trace amounts of cattle-derived materials were also detected in pig meat and bone meal and in grain-based feeds fortified with 10, 5, 1, or 0% porcine meat and bone meal. This study demonstrates the applicability of SSCP analyses to successfully identify the origin of animal species derived materials potentially present in animal feeds.