Comparison of Agar Media for Detection and Quantification of Shiga Toxin–Producing Escherichia coli in Cattle Feces

Abstract

The isolation and quantification of non-O157 Shiga toxin-producing Escherichia coli (STEC) from cattle feces are challenging. The primary objective of this study was to evaluate the performance of selected agar media in an attempt to identify an optimal medium for the detection and quantification of non-O157 STEC in cattle feces. Comparison studies were performed using CHROMagar STEC, Possé differential agar (Possé), Possé modified by the reduction or addition of antimicrobials, STEC heart infusion washed blood agar with mitomycin C (SHIBAM), and SHIBAM modified by the addition of antimicrobials. Fourteen STEC strains, two each belonging to serogroups O26, O45, O103, O111, O121, O145, and O157, were used to test detection in inoculated fecal suspensions at concentrations of 10(2) or 10(3) CFU/g. One STEC strain from each of these seven serogroups was used to estimate the concentration of recovered STEC in feces inoculated at 10(3), 10(4), or 10(5) CFU/g. Significantly more suspensions (P < 0.05) were positive for STEC when plated on Possé containing reduced concentrations of novobiocin and potassium tellurite compared with SHIBAM, but not SHIBAM modified by containing these same antimicrobials at the same concentrations. Numerically, more suspensions were positive for STEC by using this same form of modified Possé compared with Possé, but this difference was not statistically significant. More suspensions were positive for STEC cultured on CHROMagar STEC compared with those on Possé (P < 0.05) and on modified Possé (P = 0.05). Most inoculated fecal suspensions below 10(4) CFU/g of feces were underestimated or not quantifiable for the concentration of STEC by using CHROMagar STEC or modified Possé. These results suggest that CHROMagar STEC performs better than Possé or SHIBAM for detection of STEC in bovine feces, but adjustments in the concentrations of novobiocin and potassium tellurite in the latter two media result in significant improvements in their performance.