Cumulative acquisition of pathogenicity islands has shaped virulence potential and contributed to the emergence of LEE-negative Shiga toxin-producing Escherichia coli strains.

Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens causing severe gastroenteritis, which may lead to hemolytic uremic syndrome. The Locus of Enterocyte Effacement (LEE), a Pathogenicity Island (PAI), is a major determinant of intestinal epithelium attachment of a group of STEC strains; however, the virulence repertoire of STEC strains lacking LEE, has not been fully characterized. The incidence of LEE-negative STEC strains has increased in several countries, highlighting the relevance of their study. In order to gain insights into the basis for the emergence of LEE-negative STEC strains, we performed a large-scale genomic analysis of 367 strains isolated worldwide from humans, animals, food and the environment. We identified uncharacterized genomic islands, including two PAIs and one Integrative Conjugative Element. Additionally, the Locus of Adhesion and Autoaggregation (LAA) was the most prevalent PAI among LEE-negative strains and we found that it contributes to colonization of the mice intestine. Our comprehensive and rigorous comparative genomic and phylogenetic analyses suggest that the accumulative acquisition of PAIs has played an important, but currently unappreciated role, in the evolution of virulence in these strains. This study provides new knowledge on the pathogenicity of LEE-negative STEC strains and identifies molecular markers for their epidemiological surveillance.

Impacts of Climate and Management Variables on the Contamination of Preharvest Leafy Greens with Escherichia coli.

The observed seasonality of foodborne disease suggests that climatic conditions play a role and that changes in the climate may affect the presence of pathogens. However, it is hard to determine whether this effect is direct or whether it works indirectly through other factors, such as farm management. This study aimed to identify the climate and management variables that are associated with the contamination (presence and concentration) of leafy green vegetables with E. coli. This study used data about E. coli contamination from 562 leafy green vegetables (lettuce and spinach) samples taken between 2011 and 2013 from 23 open-field farms in Belgium, Brazil, Egypt, Norway, and Spain. Mixed-effect logistic and linear regression models were used to study the statistical relationship between the dependent and independent variables. Climate variables and agricultural management practices together had a systematic influence on E. coli presence and concentration. The variables important for E. coli presence included the minimum temperature of the sampling day (odds ratio = 1.47), region, and application of inorganic fertilizer. The variables important for concentration (R(2) = 0.75) were the maximum temperature during the 3 days before sampling and the region. Temperature had a stronger influence (had a significant parameter estimate and the highest R(2)) than did management practices on E. coli presence and concentration. Region was a variable that masked many management variables, including rainwater, surface water, manure, inorganic fertilizer, and spray irrigation. Climate variables had a positive relationship with E. coli presence and concentration. Temperature, irrigation water type, fertilizer type, and irrigation method should be systematically considered in future studies of fresh produce safety.