Phenotypic and Genotypic Characterization of Escherichia coli Isolated from Untreated Surface Waters

Abstract

A common member of the intestinal microbiota in humans and animals isEscherichia coli. Based on the presence of virulence factors,E. colican be potentially pathogenic. The focus of this study was to isolateE. colifrom untreated surface waters (37 sites) in Illinois and Missouri and determine phenotypic and genotypic diversity among isolates. Water samples positive for fecal coliforms based on the Colisure®test were streaked directly onto Eosin Methylene Blue (EMB) agar (37°C) or transferred to EC broth (44.5°C). EC broth cultures producing gas were then streaked onto EMB agar. Forty-five isolates were identified asE. coliusing API 20E and Enterotube II identification systems, and some phenotypic variation was observed in metabolism and fermentation. Antibiotic susceptibility of each isolate was also determined using the Kirby-Bauer Method. Differential responses to 10 antimicrobial agents were seen with 7, 16, 2, and 9 of the isolates resistant to ampicillin, cephalothin, tetracycline, and triple sulfonamide, respectively. All of the isolates were susceptible or intermediate to amoxicillin, ciprofloxacin, polymyxin B, gentamicin, imipenem, and nalidixic acid. Genotypic variation was assessed through multiplex Polymerase Chain Reaction for four virulence genes (stx1andstx2[shiga toxin],eaeA[intimin]; andhlyA[enterohemolysin]) and one housekeeping gene (uidA[β-D-glucuronidase]). Genotypic variation was observed with two of the isolates possessing the virulence gene (eaeA) for intimin. These findings increase our understanding of the diversity ofE. coliin the environment which will ultimately help in the assessment of this organism and its role in public health.