Sixty-two strains of Listeria monocytogenes isolated in Canada and Switzerland were investigated. Comparison based on molecular genotypes confirmed that strains in these two countries are genetically diverse. Interestingly strains from both countries displayed similar range of cold growth phenotypic profiles. Based on cold growth lag phase duration periods displayed in BHI at 4??C, the strains were similarly divided into groups of fast, intermediate and slow cold adaptors. Overall Swiss strains had faster exponential cold growth rates compared to Canadian strains. However gene expression analysis revealed no significant differences between fast and slow cold adapting strains in the ability to induce nine cold adaptation genes (lmo0501, cspA, cspD, gbuA, lmo0688, pgpH, sigB, sigH and sigL) in response to cold stress exposure. Neither was the presence of Stress survival islet 1 (SSI-1) analysed by PCR associated with enhanced cold adaptation. Phylogeny based on the sigL gene subdivided strains from these two countries into two major and one minor cluster. Fast cold adaptors were more frequently in one of the major clusters (cluster A), whereas slow cold adaptors were mainly in the other (cluster B). Genetic differences between these two major clusters are associated with various amino acid substitutions in the predicted SigL proteins. Compared to the EGDe type strain and most slow cold adaptors, most fast cold adaptors exhibited five identical amino acid substitutions (M90L, S203A/S203T, S304N, S315N, and I383T) in their SigL proteins. We hypothesize that these amino acid changes might be associated with SigL protein structural and functional changes that may promote differences in cold growth behaviour between L.?monocytogenes strains.
Listeria monocytogenes strains belonging to serotypes 1/2a and 4b are frequently linked to listeriosis. While inlA mutations leading to premature stop codons (PMSCs) and attenuated virulence are common in 1/2a, they are rare in serotype 4b. We observed PMSCs in 35% of L. monocytogenes isolates (n = 54) recovered from the British Columbia food supply, including serotypes 1/2a (30%), 1/2c (100%), and 3a (100%), and a 3-codon deletion (amino acid positions 738 to 740) seen in 57% of 4b isolates from fish-processing facilities. Caco-2 invasion assays showed that two isolates with the deletion were significantly more invasive than EGD-SmR (P 200 h) adaptors. Intermediate CAG strains (70%) more frequently possessed inlA PMSCs than did fast (20%) and slow (10%) CAGs; in contrast, 87% of fast adaptors lacked inlA PMSCs. In conclusion, we report food chain-derived 1/2a and 4b serotypes with a 3-codon deletion possessing invasive behavior and the novel association of inlA genotypes encoding a full-length InlA with fast cold-adaptation phenotypes.
In British Columbia (BC), Canada, food processing facilities licensed under provincial authority are not required to sample for Listeria monocytogenes in food products or processing environments. In 2009, we conducted a survey of dairy, fish, and meat facilities under BC authority to estimate the prevalence of Listeria spp. and L. monocytogenes in ready-to-eat (RTE) foods and production environments. From August to October, 250 RTE food samples and 258 swabs from the food processing environments of 43 facilities were collected. Standard culture methods were applied to both food samples and swabs. Of swabs collected from all 258 environmental surfaces, 15% were positive for Listeria spp. Significantly (P, 0.001) more fish facilities than dairy and meat facilities had food contact surfaces contaminated with Listeria spp. L. monocytogenes was found in RTE foods from fish facilities alone (5 of 12); in all five of the fish facilities with contaminated product, one or more environmental swabs were also positive for L. monocytogenes. The results suggest that while control of L. monocytogenes in BC-inspected dairy and meat facilities is effective in limiting food contamination, there is a need for provincial inspectors to initiate improved monitoring and management of contamination by L. monocytogenes in RTE fish processing facilities.