maltophilia. The elucidation of molecular mechanisms underlying these phenotypic differences might be relevant to the identification of new targets for designing rational and effective methods to combat and eradicate S. maltophilia infection. Methods Bacterial isolates and growth conditions Overall, 98 S. maltophilia isolates were investigated: 41 strains collected

from the sputa of CF Selleck YH25448 patients attending the CF Unit at “”Bambino Gesù”" Children’s Hospital and Research Institute of Rome; TEW-7197 ic50 47 strains collected from different sites (30 from respiratory tract, 10 from blood, and 7 from swabs) in non-CF patients attending “”Bambino Gesù”" Children Hospital of Rome, or “”Spirito Santo”" Hospital of Pe scara; and 10 strains (ENV) isolated in Czech Republic from several environmental sources (paddy, soil, rhizosphere tuberous roots, and waste water). Since in severely ill chronic obstructive pulmonary disease (COPD) patients P. aeruginosa clones similar to those in CF persists [52], patients with COPD were not enrolled AZD6094 cell line in the present study. All clinical isolates represented non-consecutive strains isolated from different patients, except for 2 CF patients with 7 and 3 isolates, respectively. The isolates were identified as S. maltophilia by biochemical tests using manual (API 20-NE System; BioMérieux, Marcy-L’Etoile,

France) or automated (Vitek; BioMérieux) systems, then stored at -80°C until use when they were grown at 37°C (and also at 25°C, in the case of ENV strains) in Trypticase Soy broth

(TSB; Oxoid SpA; Garbagnate M.se, Milan, Italy) or Mueller-Hinton agar (MHA; Oxoid) plates unless otherwise noted. Genetic relatedness by PFGE and cluster analysis After digestion of DNA with the Suplatast tosilate restriction enzyme XbaI as previously described [24, 27, 28], PFGE was carried out as follows: initial switch time and final switch time were 5 and 35 sec, respectively; DNA fragments were run with a temperature of 12°C for 20 h at 6.0 V/cm with an included angle of 120°. Isolates with identical PFGE patterns were assigned to the same PFGE type and subtype. Isolates differing by one to three bands were assigned to different PFGE subtypes but to the same PFGE type and were considered genetically related. Isolates with PFGE patterns differing by more than 4 bands were considered genetically unrelated and were assigned to different PFGE types. PFGE types were analyzed with BioNumerics software for Windows (version 2.5; Applied Maths, Ghent, Belgium). The DNA banding patterns were normalized with bacteriophage lambda concatemer ladder standards. Comparison of the banding patterns was performed by the UPGMA and with the Dice similarity coefficient. A tolerance of 1.