Lastly, a 10 days incubation of your samples on SSMS followed from the spreading from the microcolonies on solid media allowed the culture of isolates belonging to four genera. Between them, 3 were detected only on LB plates. These strains belong on the genera Bacillus and Paenibacillus from your phylum Firmicutes and Micrococcus from the phylum Actinobacteria. The advantage of this system was here to prevent fungal con tamination in comparison with direct spreading for which dif ferent moulds invaded the LB plates, regardless of the presence of antifungal agents. Acidocella have been also detected after growth on SSMS and spreading on FD1 media. They shared an identical 16S rRNA gene sequence with strains isolated directly on FD media. Our outcomes indicate the isolated strains are remarkably unique for the medium applied for principal isolation, since the majority of them were found on just one medium.
As previously stated, exceptions are located for members in the genus Cellulomonas sp. which have been iso lated on both FD1 and FD2 media and Acidocella sp. which had been uncovered on FD1, FD2 and FD4 media. Isolation of previously uncultured bacteria The culture strategy led here towards the isolation discover this of repre sentatives of three new genera as defined by Tindall et al, Indeed, these three isolates shared under 95% 16S rRNA gene sequence identity with their closest taxo nomically characterized species. The very first 1 belongs on the phylum Acidobacteria and shared 98% 16S rRNA gene sequence identity with uncultured clone IRON SNOW NB E11 whereas its closest taxonomically characterized species was Acidobacterium capsulatum with 94% identity.
Acidobacteria correspond to some of one of the most abundant microorgan isms in the setting but are recalcitrant to cultivate in laboratory, Only eight genera from this phylum have already been taxonomically selleck isolated thus far, The recent metagenomic investigation of Carnoul?s AMD, led to the detection of a strain of Acidobacteria i. e. CARN3, However, N3B showed lower than 95% 16S identity with CARN3. The second 1 belongs for the fam ily Xanthomonadaceae. X11 sequence was closely associated with the uncultured Proteobacterium DKE however the closest taxonomically character ized strain was Dokdonella koreensis with less than 94% identity. X11 and its closest associated strains are frequent inhabitants of mines and acidic environ ments, given that BLAST evaluation revealed the presence of their 16S rDNA in Carnoul?s likewise as in a variety of other acid mine drainages and acidic waters, The third a single is H7p, a member from the Propionibacter iaceae family members. Its finest matching 16S rRNA gene sequence is Luteococcus sp. uncovered in human peritoneum sharing only 95% identity with H7p. No superior identity was discovered in any database even with uncultured bacteria .