Molecular Typing of Potentially Pathogenic Escherichia coli Isolated from Fresh Pasta Filata Venezuelan Cheeses

Abstract

Background. Fresh pasta filata cheese is considered as one of the most important foods in the Venezuelan diet. It is typically produced by small-scale producers using raw milk. The objective of this research was to molecularly characterize the pathogenic potential of Escherichia coli strains isolated from pasta filata cheese manufactured and marketed in Venezuela. Methodology. In the period between January and March of 2019, a total of 36 strains of E. coli were isolated from a variety of pasta filata cheeses including 17 samples of mozzarella, 16 of telita, and 3 of guayanés. These strains were isolated according to the Venezuelan Commission of Industrial Standards (COVENIN) and identified by conventional methods (biochemical and phenotypic tests). Antimicrobial susceptibility was determined using the disk diffusion technique. Phylogenetic grouping and detection of virulence genes were performed by Polymerase Chain Reaction amplification. Diversity and genetic relationships were determined by Rep-PCR. Results: All strains were susceptible to the tested antibiotics. Phylogroup A (n=19) was the most frequent (52.8%), followed by groups D (n=11; 30.6%), and B1 (n=2; 5.6%). The majority of isolates carried at least two virulence genes, one coding for adhesion mechanisms (fimH) and the other for iron uptake (fyuA). Only one strain of phylogroup A presented a profile consisting of four virulence genes (fimH, fyuA, kpsMT II, and papAH). Four strains that could not be classified according to Clermont's scheme carried resistance genes as well. A heterogeneous population structure was observed by Rep-PCR of the strains. Conclusion: Results support the hypothesis that the E. coli strains isolated from the three types of pasta filata cheeses manufactured and marketed in Venezuela have identical characteristics and virulence factors to Extraintestinal Pathogenic E. coli strains observed in animals and humans, posing a potential health risk. Therefore, it is essential to improve hygienic and sanitary controls at all stages of cheese production and to implement measures for epidemiological surveillance of potentially pathogenic bacterial strains present in Venezuelan, artisanal pasta filata cheeses.