title: Value of lysozyme agar incorporation and alkaline thioglycollate exposure for the environmental recovery of Clostridium difficile.
abstract: Clostridium difficile is an increasingly prevalent nosocomial pathogen. Environmental contamination by spores is believed to be a major factor propagating the spread of C. difficile. Various approaches including the use of bile salts have been described to enhance the recovery of C. difficile from clinical and environmental specimens. We found that lysozyme (5 mg/L) incorporated into a selective medium containing bile salts significantly increased the recovery of C. difficile from swabs of 197 environmental sites (11% versus 24% samples positive, P< 0.01). Furthermore, in a separate series of experiments additional use of cooked meat broth enrichment significantly enhanced the recovery of C. difficile (35% versus 45%, P = 0.009). Conversely, we found that pre-exposure to alkaline thioglycollate did not improve the yield of C. difficile. Lysozyme incorporation markedly increases the recovery of C. difficile from environmental samples probably by stimulation of spore germination. Our findings suggest that previous attempts to determine the level of environmental C. difficile contamination have markedly underestimated the true prevalence of this pathogen.
authors.affiliation: Department of Microbiology, University of Leeds and The General Infirmary, Leeds, LS2 9JT, UK.
authors.forename: M H
authors.initials: MH
authors.lastname: Wilcox
authors.forename: W N
authors.initials: WN
authors.lastname: Fawley
authors.forename: P
authors.initials: P
authors.lastname: Parnell
mesh_terms: D000362:Agar; D000900:Anti-Bacterial Agents; D001431:Bacteriological Techniques / Q000706:statistics & numerical data; D001647:Bile Acids and Salts; D016009:Chi-Square Distribution; D016360:Clostridioides difficile / Q000302:isolation & purification*; D003470:Culture Media; D004783:Environmental Microbiology; D009113:Muramidase; D013864:Thioglycolates
chemical_list: D000900:Anti-Bacterial Agents; D001647:Bile Acids and Salts; D003470:Culture Media; D013864:Thioglycolates; D000362:Agar; D009113:Muramidase