Characterization of aquatic clade 2 and 3 Campylobacter coli isolates from Slovenia reveals admixture with other Campylobacter species

Campylobacter coli, a significant foodborne pathogen, has undergone extensive genetic exchange with its close relative, Campylobacter jejuni, leading to the emergence of three distinct clades. While clade 1 strains are commonly isolated from clinical and agricultural sources, clades 2 and 3 are primarily found in aquatic environments. This study aimed to enhance our understanding of C. coli clade 2 and 3 isolates through genomic and phenotypic characterization. A total of 48 surface water samples were collected from 19 different water bodies throughout Slovenia, and eleven Campylobacter isolates initially identified as C. coli from clades 2 and 3 were cultured. Whole genome sequencing was then performed on these isolates. Phylogenetic analysis was conducted using core genome multilocus sequence typing (cgMLST) and k-mer analysis. Phenotypic characterization included growth analysis, autoagglutination, biofilm formation, motility, antimicrobial susceptibility, water survival, and metabolic profiling. Genomic analysis revealed significant admixture with other Campylobacter species in the clade 2 and 3 isolates. One isolate was found to represent a new species related to C. coli. Besides C. jejuni and C. lari, this novel species appears to have contributed to introgression in the C. coli clades 2 and 3 isolates. Phenotypic characterization demonstrated diverse growth patterns, motility, autoagglutination abilities, and biofilm formation among the isolates. This study provides new insights into the genetic diversity and phenotypic characteristics of aquatic C. coli clade 2 and 3 isolates from Slovenia. The observed admixture with other Campylobacter species highlights the complex evolutionary history of these environmental strains and underscores the importance of continued surveillance and characterization of Campylobacter isolates from diverse ecological niches.