Isolation and characterization of potentially pathogenic antimicrobial-resistant Escherichia coli strains from chicken and pig farms in Spain

Abstract

To ascertain whether on animal farms there reside extended-spectrum beta-lactamase (ESBL) and plasmidic class C beta-lactamase-producing Escherichia coli isolates potentially pathogenic for humans, phylogenetic analyses, pulsed-field gel electrophoresis (PFGE) typing, serotyping, and virulence genotyping were performed for 86 isolates from poultry (57 isolates) and pig (29 isolates) farms. E. coli isolates from poultry farms carried genes encoding enzymes of the CTX-M-9 group as well as CMY-2, whereas those from pig farms mainly carried genes encoding CTX-M-1 enzymes. Poultry and pig isolates differed significantly in their phylogenetic group assignments, with phylogroup A predominating in pig isolates and phylogroup D predominating in avian isolates. Among the 86 farm isolates, 23 (26.7%) carried two or more virulence genes typical of extraintestinal pathogenic E. coli (ExPEC). Of these, 20 were isolated from poultry farms and only 3 from pig farms. Ten of the 23 isolates belonged to the classic human ExPEC serotypes O2:H6, O2:HNM, O2:H7, O15:H1, and O25:H4. Despite the high diversity of serotypes and pulsotypes detected among the 86 farm isolates, 13 PFGE clusters were identified. Four of these clusters contained isolates with two or more virulence genes, and two clusters exhibited the classic human ExPEC serotypes O2:HNM (ST10) and O2:H6 (ST115). Although O2:HNM and O2:H6 isolates of human and animal origins differed with respect to their virulence genes and PFGE pulsotypes, the O2:HNM isolates from pigs showed the same sequence type (ST10) as those from humans. The single avian O15:H1 isolate was compared with human clinical isolates of this serotype. Although all were found to belong to phylogroup D and shared the same virulence gene profile, they differed in their sequence types (ST362-avian and ST393-human) and PFGE pulsotypes. Noteworthy was the detection, for the first time, in poultry farms of the clonal groups O25b:H4-ST131-B2, producing CTX-M-9, and O25a-ST648-D, producing CTX-M-32. The virulence genes and PFGE profiles of these two groups were very similar to those of clinical human isolates. While further studies are required to determine the true zoonotic potential of these clonal groups, our results emphasize the zoonotic risk posed especially by poultry farms, but also by pig farms, as reservoirs of ESBL- and CMY-2-encoding E. coli.

FIG. 1.

XbaI PFGE dendrogram of 13 clusters (similarity, ≥85%) including 33 isolates of poultry and porcine origins. The dendrogram was produced by the UPGMA algorithm based on a Dice similarity coefficient with a 1.0% band position tolerance (Tol).

FIG. 2.

XbaI PFGE dendrogram of 13 isolates belonging to serotypes O2:HNM and O2:H6 of human and farm origins. The dendrogram was produced by the UPGMA algorithm based on a Dice similarity coefficient with a 1.0% band position tolerance.

FIG. 3.

XbaI PFGE dendrogram of five isolates belonging to serotypes O15:H1 and O25a:H1 of human and farm origins. The dendrogram was produced by the UPGMA algorithm based on a Dice similarity coefficient with a 1.0% band position tolerance.

FIG. 4.

XbaI-PFGE dendrogram of 21 isolates belonging to O25 serogroup of human and farm origin. The dendrogram was produced by the UPGMA algorithm based on a Dice similarity coefficient with a 1.0% band position tolerance.