[25]. The sequence comparison of this gene has been already used for species identification and phylogenetic analysis of other genera (e.g. Staphylococcus, Lactobacillus) and enteric pathogens [26–28]. A chaperonin database (cpnDB) is available on line, collecting Selonsertib datasheet bacterial and eukaryotic sequences (http://​www.​cpndb.​ca/​cpnDB/​home.​php)

[29]. The purpose of this study is the development of a rapid, reproducible and easy-to-handle molecular tool for the identification of Bifidobacterium species isolated from various environments. The protocol is based on the restriction endonuclease analysis of the PCR-amplified hsp60 partial gene sequence (hsp60 PCR-RFLP) with the use of a single restriction enzyme and has been tested on the 30 most widely distributed Bifidobacterium TGF-beta inhibitor species and subspecies. PHA-848125 A diagnostic dichotomous key to speed up profile interpretation has also been proposed. Methods Bacterial strains and culture conditions The type strains used to develop the technique are listed in Table  1, whereas the strains used to validate the method are reported in Table  2. The strains, belonging to BUSCoB (Bologna University Scardovi Collection of Bifidobacteria) collection, were isolated from faeces

of human and animals and from sewage. Bacteria were maintained as frozen stocks at −80°C in the presence of skim milk as cryoprotective agent. Working cultures were prepared in TPY medium [1], grown anaerobically at 37°C and harvested at logarithmic phase. Table 1 Type-strains investigated Species International culture collection Bifidobacterium adolescentis ATCC 15703 Bifidobacterium angulatum ATCC 27535 Bifidobacterium animalis subsp. animalis ATCC 25527 Bifidobacterium animalis subsp. lactis DSM 10140 Bifidobacterium asteroides ATCC 25910 Bifidobacterium bifidum ATCC 29521 Bifidobacterium boum ATCC 27917 Bifidobacterium breve ATCC 15700 Bifidobacterium catenulatum ATCC 27539 Bifidobacterium choerinum ATCC 27686 Bifidobacterium

coryneforme ATCC 25911 Bifidobacterium cuniculi ATCC 27916 Bifidobacterium dentium ATCC this website 27534 Bifidobacterium gallicum ATCC 49850 Bifidobacterium gallinarum ATCC 33777 Bifidobacterium indicum ATCC 25912 Bifidobacterium longum subsp. longum ATCC 15707 Bifidobacterium longum subsp. infantis ATCC 15697 Bifidobacterium longum subsp. suis ATCC 27533 Bifidobacterium minimum ATCC 27539 Bifidobacterium merycicum ATCC 49391 Bifidobacterium pseudolongum subsp pseudolongum ATCC 25526 Bifidobacterium pseudolongum subsp. globosum ATCC 25865 Bifidobacterium pseudocatenulatum ATCC 27919 Bifidobacterium pullorum ATCC 27685 Bifidobacterium ruminantium ATCC 49390 Bifidobacterium subtile ATCC 27537 Bifidobacterium thermacidophilum subsp. porcinum LMG 21689 Bifidobacterium thermacidophilum subsp.