Strategies for recovering of planktonic and sessile cells of Escherichia coli O157:H7 from freshwater environment

Abstract

The experiments were performed with Escherichia coli O157:H7 EDL 933 in freshwater microcosms at 12 °C. At 35, 45, and 70 days, samples were taken and filtered through 0.45 μm membrane filters. The following alternatives were tested to evaluate the recovery percentage of injured cells: (1) selective media CHROMagar(™)O157 and chromID(™)O157:H7 agar, at 37 °C for 24 h; (2) tryptic soy agar supplemented with yeast extract (TSAE), incubated at 25 °C for 2 or 4 h, then transferred to CHROMagar(™)O157 or chromID(™)O157:H7 agar at 37 °C (TSAE2h-CHROM, TSAE4h-CHROM and TSAE2h-ID, TSAE4h-ID); (3) thin agar layer (TAL) method, TSAE was overlaid on CHROMagar(™)O157 or chromID(™)O157:H7 agar (TALCHROM and TALID, respectively) and incubated at 37 °C for 24 h; and (4) TALCHROM at 25 °C for 4 h, then continued up to complete 24 h at 37 °C (TALCHROM4h). Furthermore, the recovery of E. coli O157:H7 cells adhering to glass coverslips were evaluated to mimic biofilm conditions. The recovery percentages obtained from each alternative were calculated relative to TSAE counts. After 70 days, TSAE4h-CHROM and TALCHROM4h showed the highest recovery percentage (>90 %) from water microcosms. Despite the improved recovery of cell adhering to glass surfaces, the percentages obtained with TSAE4h-CHROM were low. Further studies for the recovery of biofilm-forming E. coli O157:H7 are required. Pre-incubation on TSAE at 25 °C for 4 h, combined with CHROMagar(™)O157, or by thin agar layer method (TALCHROM) enhanced significantly the recovery of viable cells of E. coli O157:H7 after prolonged stay in water microcosms.

Application of atmospheric cold plasma, ultraviolet radiation, or natural antimicrobials for control of foodborne pathogenic and spoilage microorganisms

“…It can easily differentiate targeted microbes from the background microflora and at the same time recover cells sub-lethally injured by UV radiation. In a study evaluating the strategies for recovering sublethally injured E. coli O157:H7 after 70 days in freshwater microcosms, the TAL method (TSAYE overlaid on CHROMagar ™ O157), and TSAYE pre-incubated at 25 °C for 4 h then transferred to CHROMagar ™ O157 at 37 °C, recovered the highest percentage (>90%) of viable E. coli O157:H7 (Marucci & Cubitto, 2016).…”