16 In the current study, AFLP was found to be useful for discrimination between inter- and intrapatient isolates. Moreover, GSK126 in vivo all isolates could be identified down to the species

level according to the current taxonomic status. A majority of patients were exclusively colonised by one AFLP genotype. Only one genotype was shared between two patients. The colonisation of CF patients by multiple AFLP genotypes was already reported previously [37] but this study was performed in 2002, well before the recent taxonomical changes. Therefore, from the present perspective, we cannot appraise if intra- or interspecific variations were detected. Defontaine et al.37 state multiple colonisations with up to three different genotypes, comprising one predominant genotype associated with up to two accompanying genotypes. Exceptionally, in our study, we found patients colonised with up to five different genotypes over a period of up to 5 years, with re-appearing genotypes. Therefore,

it is very likely that those patients are colonised with multiple S. prolificans genotypes. Our data mirror that CF patients can be chronically colonised with a specific genotype or multiple genotypes for prolonged periods of time (several years). Co-colonisation by multiple genotypes, also in non-CF patients, has been recognised before for other fungal species, such as A. fumigatus and A. flavus.36,38,39 If multiple colonisation turns into multiple infections by different genotypes of one species, this might have an impact on disease outcome, check details as we found that different Scedosporium isolates from the same patient (Table 1) can vary considerably in their AFSP. In particular, when patients are colonised by two or more Megestrol Acetate isolates with different susceptibility patterns, this may result in an overestimation of MIC values. This situation is exemplified in this study for instance in patient 13 where one clinical sample contained an MICA-susceptible, as well as MICA-resistant isolate of the same species. Apparently,

also patient 1 was colonised at the same time with two isolates of the same species, but with different AFSPs. For this reason, clinical specimens should be carefully analysed for the possible presence of multiple strains expressing variable antifungal susceptibilities. Overseeing such mixed infections due to S. prolificans may in part explain the therapy refractive nature of S. prolificans. In conclusion, we found that S. prolificans represents the most prevalent Scedosporium species in the respiratory tract of CF patients and immunocompromised patients in Northern Spain. In CF patients, P. boydii or S. prolificans were exclusively found as respiratory colonisers. All patients were colonised over years exclusively with isolates affiliated to one Scedosporium species, but to multiple AFLP genotypes carrying variable AFSP.