Clostridium difficile is a common and potentially fatal cause of antibiotic-associated diarrhoea and pseudomembranous colitis worldwide. It has been isolated from patients and their surroundings, in healthcare facilities and from the community. C. difficile is able to survive for many months on inanimate surfaces in the form of spores. PCR ribotyping is used in the UK to characterise and identify strain diversity. Investigating how the most problematic strains respond to cleaning regimes may influence the control of disease. This work used the University Hospitals of Leicester Trust as a case study for this purpose of understanding the epidemiology of this pathogen within healthcare facilities. Five individual agar media were compared based on their abilities to recover and resuscitate damaged ribotype 027 spores, a strain associated with disease outbreaks and increased severity. Controlled laboratory experiments with a sub-lethal dose of a germicide were conducted before C. difficile recovery from hospital wards. An additional two sampling campaigns acquired environmental strains. C. difficile isolation after routine cleaning demonstrated the inefficiency of the current recovery regime as C. difficile spores were recovered using direct contact plates, enrichment broths, and resuscitation media. This study used layering of non-selective agar over selective agar, identifying a potential link in the proportions of media following the use of sponges in environmental sampling. All strains were characterised by ribotyping; ribotype 027 was isolated from all sampling cohorts. A four-month epidemiological study was conducted into the ribotype prevalence and distribution from C. difficile-positive faecal specimens. A second survey investigated these effects with a modification of C. difficile detection from faecal samples. Hydrogen peroxide vapour is currently being explored as a means of decontamination of healthcare-associated infections. Inactivation kinetics of ribotype 027 spores were analysed in response to vapour and liquid exposure of hydrogen peroxide. No reports thus far have explored such kinetics and controlled decontamination with both clinical and non-clinical strains. Evidence strongly suggests spores can be inactivated with its application. Furthermore, this study revealed there appears to be significant differences in susceptibility and inactivation of different C. difficile ribotypes.
A Doctoral Thesis. Submitted in partial fulfilment of the requirements for the award of Doctor of Philosophy of Loughborough University.