Occurrence of extended-spectrum β-lactamase-producing bacteria in urban Clinton River habitat

The aim of this study was to determine whether Clinton River water is contaminated with antibiotics and is a reservoir of antimicrobial-resistant bacteria.

Water samples were taken from two sites of Clinton River. Antimicrobial-resistant bacteria were enumerated on agar plates supplemented with six commonly used antibiotics. Extended-spectrum β-lactamase (ESBL)-producing bacteria were identified using a BD Phoenix™ System and by 16S rRNA gene sequencing. Antimicrobial resistance gene transfer was performed by conjugation studies and the location of genes was determined by Southern hybridisation. Virulence properties of ESBL-producing isolates were determined by assessing their biofilm-forming ability, cellular toxicity, and induction of an inflammatory response in intestinal epithelial (Caco-2) cells.

16S rRNA analysis of water samples showed the presence of potentially pathogenic bacteria (e.g. Shigella flexneri, Klebsiella pneumoniae, Aeromonas punctata and Pseudomonas aeruginosa). Among 64 biochemically identified bacterial isolates tested, 42% were resistant to cefotaxime, 34% to chloramphenicol, 9% to tetracycline, 11% to ciprofloxacin and 9% to gentamicin. Of 27 cefotaxime-resistant isolates, 11 (41%) were ESBL-positive and possessed either bla_CTX-M (n=9), bla_TEM (n=1) or bla_KPC (n=1). Comparative analysis of ESBL gene sequences from Clinton River water bacteria showed 98-100% identity with clinical isolates. ESBL-producing isolates from Clinton River water were found to form biofilms, induced inflammatory cytokines and caused toxicity to epithelial cells.

Clinton River water contains isolates with ESBL genes identical to clinical isolates and possessing virulence properties, thus it could be a potential reservoir in causing human infections.