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Aim: To determine the prevalence of Burkholderia cepacia from the environment in a regional adult cystic fibrosis (CF) care centre., Methods: Two hundred and eighty-five environmental samples were taken from inpatient and outpatient environments within the CF centre. Following a non-selective enrichment step, broths were plated onto a selective medium and B. cepacia isolates were confirmed by phenotypic tests and also genotypically by species specific 16S rRNA PCR., Results: B. cepacia was not detected from commonly shared items of equipment, staff hands, staff uniforms or toilets. In addition, the organism was not detected in toilet bowls, even in the B. cepacia unit. With regard to positive environments for B. cepacia, 4/10 (40%) of the outside surfaces and inner rims of patients' plastic disposable sputum collection containers and 4/17 (23.5%) of air from patients' rooms, following physiotherapy, were positive., Conclusions: All positive samples originated in the B. cepacia segregation area of the inpatient wards and B. cepacia was not detected in the non-cepacia area of the CF centre. Consequently, these two positive sites should therefore be treated as high risk, where organisms may be potentially transmitted from environment to patient. As there is now evidence to suggest differences in the virulence and hence clinical outcome of certain of the genomovars of this organism, this study may be important in highlighting the areas that patients who are B. cepacia positive should avoid, to minimise the risk of transmission and acquisition of this organism within the CF cepacia population. These results indicate that effective infection control procedures are required in patients' rooms following physiotherapy, for prevention of B. cepacia transmission and to avoid the transmission of different genomovar types within CF patients, who are already colonised with this pathogen.