Transcription levels of hes and their involvement in the biofilm formation of Shiga toxin-producing Escherichia coli O91.

Shiga toxin-producing Escherichia coli (STEC) are recognized as being responsible for many cases of foodborne diseases worldwide. Cattle are the main reservoir of STEC, shedding the microorganisms in their feces. The serogroup STEC O91 has been associated with hemorrhagic colitis and hemolytic uremic syndrome. Locus of Adhesion and Autoaggregation (LAA) and its hes gene are related to the pathogenicity of STEC and the ability to form biofilms. Considering the frequent isolation of STEC O91, the biofilm-forming ability, and the possible role of hes in the pathogenicity of STEC, we propose to evaluate the ability of STEC to form biofilms and to evaluate the expression of hes before and after of biofilm formation. All strains were classified as strong biofilm-forming. The hes expression showed variability between strains before and after biofilm formation, and this may be due to other genes carried by each strain. This study is the first to report the relationship between biofilm formation, and hes expression and proposes that the analysis and diagnosis of LAA, especially hes as STEC O91 virulence factors, could elucidate these unknown mechanisms. Considering that there is no specific treatment for HUS, only supportive care, it is necessary to know the survival and virulence mechanisms of STEC O91.

Distribution of Locus of Adhesion and Autoaggregation and hes Gene in STEC Strains from Countries of Latin America.

Shiga toxin-producing Escherichia coli (STEC) are zoonotic food pathogens associated with foodborne diarrheal illness, hemorrhagic colitis, and complications such as hemolytic uremic syndrome (HUS). The ability to adhere to epithelial cells is an important virulence trait, and pathogenicity islands (PAIs) play an important role on it. Some STEC carrying a PAI named locus of enterocyte effacement (LEE-positive) have been frequently associated to HUS; however, STEC that do not carry LEE (LEE-negative) have also been associated with this outcome. The burden of disease caused by LEE-negative STEC has increased recently in several countries like Argentina, Chile, and Paraguay. A new PAI -the Locus of Adhesion and Autoagregation (LAA)-has been associated to severe disease in humans. In this study, we aimed to analyze the distribution of LAA and its possible predictor, the gene hes, in LEE-negative STEC strains isolated from Chile and Paraguay from different sources. The presence of the different LAA modules and hes were detected by PCR. LAA was found in 41.6% and 41.0% of strains isolated from Chile and Paraguay, respectively. Strains were isolated from diverse origins and belonged to several serogroups including O91, O103, and O113. The hes gene was detected in 50% of the isolates from Paraguay and Chile. Therefore, the detection of LAA and hes in STEC could complement current genetic evaluation schemes, allowing to classify LEE negative STEC strains as LAA-positive or LAA-negative STEC strains.

Identification and detection of iha subtypes in LEE-negative Shiga toxin-producing Escherichia coli (STEC) strains isolated from humans, cattle and food.

LEE-negative Shiga toxin-producing Escherichia coli (STEC) strains are important cause of infection in humans and they should be included in the public health surveillance systems. Some isolates have been associated with haemolytic uremic syndrome (HUS) but the mechanisms of pathogenicity are is a field continuos broadening of knowledge. The IrgA homologue adhesin (Iha), encoded by iha, is an adherence-conferring protein and also a siderophore receptor distributed among LEE-negative STEC strains. This study reports the presence of different subtypes of iha in LEE-negative STEC strains. We used genomic analyses to design PCR assays for detecting each of the different iha subtypes and also, all the subtypes simultaneously. LEE-negative STEC strains were designed and different localizations of this gene in STEC subgroups were examinated. Genomic analysis detected iha in a high percentage of LEE-negative STEC strains. These strains generally carried iha sequences similar to those harbored by the Locus of Adhesion and Autoaggregation (LAA) or by the plasmid pO113. Besides, almost half of the strains carried both subtypes. Similar results were observed by PCR, detecting iha LAA in 87% of the strains (117/135) and iha pO113 in 32% of strains (43/135). Thus, we designed PCR assays that allow rapid detection of iha subtypes harbored by LEE-negative strains. These results highlight the need to investigate the individual and orchestrated role of virulence genes that determine the STEC capacity of causing serious disease, which would allow for identification of target candidates to develop therapies against HUS.

First report of the distribution of Locus of Adhesion and Autoaggregation (LAA) pathogenicity island in LEE-negative Shiga toxin-producing Escherichia coli isolates from Argentina.

Shiga toxin-producing Escherichia coli (STEC) are important foodborne pathogens that can cause severe disease. The ability to adhere to epithelial cells is an important virulence trait and pathogenicity islands (PAIs) play an important role. Recently, researchers identified a member of the Heat-resistant agglutinin family and characterized this antigen named Hemagglutinin from Shiga toxin-producing E. coli (Hes). More importantly, they showed that hes and other genes such as iha, pagC and agn43 were integrated in each of the four modules present in the new PAI named Locus of Adhesion and Autoaggregation (LAA) whose presence is associated with severe disease linked to with LEE-negatives STEC. The distribution of LAA among STEC strains isolates from different origins between 2000 and 2015 from cattle, the farm environment, and food and harboring diverse virulence was investigated. The STEC strains were characterized by PCR to detect three modules of LAA and agn43 (as marker of module IV), and phylogenetic groups were determined. LAA was found in 46% of LEE-negative STEC corresponding to serogroups O91, O174, O113, O171, O178, O130 and others. The presence of this PAI is associated with strains harboring stx2 (56%) and belonging to phylogroup B1 (91%). LAA is a novel pathogenicity island associated with strains isolated from Hemolytic Uremic Syndrome cases. Therefore, the results of this study contribute to a better understanding regarding the pathogenicity of this emergent subset of STEC strains harboring LAA as a predictor of virulence of LEE-negative STEC strains.