Exploring the epidemiology and genetic complexity of Escherichia coli in vertically integrated pig production systems using whole genome sequencing

Escherichia coli is a major pathogen responsible for enteric diseases in swine. This study aims to assess the genetic diversity, virulence factors, and antimicrobial resistance genes of E. coli isolated from diarrheic pigs within Spanish integrated production systems. Metadata, including epidemiological data and minimum inhibitory concentration (MIC), was collected for 350 clinical isolates reported to the laboratory between 2018 and 2022. MIC analysis using Ward's hierarchical clustering identified 17 phenotypic resistance clusters. A subset of 206 isolates, representing different production pyramids and phenotypic clusters, was selected for whole genome sequencing (WGS). In silico serotyping and multilocus sequence typing revealed high genetic diversity, with predominant serotypes O138:H14, O147:H14, O141:H4, and O108:H39, and sequence types ST10 and ST42. Only three ST131 isolates were found in a single production pyramid suggesting that swine associated ST131 is not contributing significantly to the epidemiology of human infections associated to this ST. Isolates were classified into six phylogroups, with phylogroup A being the most represented. AMR profiling identified a high percentage of resistance (> 50 %) to tetracyclines, aminoglycosides, quinolones, trimethoprim and sulphonamides, while resistance to third-generation cephalosporins and colistin remained low (< 25 %). Comparative genomic analysis demonstrated high genetic diversity within production pyramids, although, some clones persisted over time, suggesting transmission within batches. Statistical analyses found no association between genetic traits and farm origins, although outbreaks often involved clones with minimal SNP differences. This study underscores the complexity of E. coli transmission in swine and highlights the need for genomic surveillance to monitor AMR.

This study was partially funded by AEMPS / PRAN with the European funds \u201CMecanismo de recuperaci\u00F3n y resilencia ( C18.I2.P3.S2 )\u201D and the CERCA program. Anna Vilar\u00F3 is supported by the Ag\u00E8ncia de Gesti\u00F3 d\u2019Ajuts Universitaris i de Recerca ( AGAUR ) of the Catalan Government though the industrial doctorate grant 2021DI 86 . This study was carried out with the support of Grup de Sanejament Porc\u00ED, Lleida (GSP).