TL;DR: The 16S rRNA gene sequences were determined by PCR direct sequencing and were compared with more than 80 previously determined clostridial sequences and the previously published sequences of representative species of other low- G + C-content gram-positive genera, thereby providing an almost complete picture of the genealogical interrelationships of theClostridia.
Abstract: The 16S rRNA gene sequences of 34 named and unnamed clostridial strains were determined by PCR direct sequencing and were compared with more than 80 previously determined clostridial sequences and the previously published sequences of representative species of other low- G+C-content gram-positive genera, thereby providing an almost complete picture of the genealogical interrelationships of the clostridia. The results of our phylogenetic analysis corroborate and extend previous findings in showing that the genus Clostridium is extremely heterogeneous, with many species phylogenetically intermixed with other sporeforming and non-sporeforming genera. The genus Clostridium is clearly in need of major revision, and the rRNA structures defined in this and previous studies may provide a sound basis for future taxonomic restructuring. The problems and different possibilities for restructuring are discussed in light of the phenotypic and phylogenetic data, and a possible hierarchical structure for the clostridia and their close relatives is presented. On the basis of phenotypic criteria and the results of phylogenetic analyses the following five new genera and 11 new combinations are proposed: Caloramator gen. nov., with Caloramator fervidus comb. nov.; Filifactor gen. nov., with Filifactor villosus comb. nov.; Moorella gen. nov., with Moorella thermoacetica comb. nov. and Moorella thermoautotrophica comb. nov.; Oxobacter gen. nov., with Oxobacter pfennigii comb. nov.; Oxalophagus gen. nov., with Oxalophagus oxalicus comb. nov.; Eubacterium barkeri comb. nov.; Paenibacillus durum comb. nov.; Thermoanaerobacter kivui comb. nov.; Thermoanaerobacter thermocopriae comb. nov.; and Thermoanerobacterium thermosaccharolyticum comb. nov.
TL;DR: The human pharyngeal microbiome of healthy individuals in Greece was characterization and unique differentially abundant families, genera and species were identified, and further studies are needed to elucidate their role.
Abstract: BACKGROUND The recent advent of high-throughput sequencing methods enabled the study of the composition of the upper respiratory tract (URT) microbial ecosystem and its relationship with health and disease in immense detail. The aim of the present study was the characterization of the human pharyngeal microbiome of healthy individuals in Greece. MATERIALS AND METHODS We obtained ten pharyngeal specimens from healthy volunteers, Greek resident, with Greek nationality, who were eligible to the selection criteria. The construction of DNA libraries was performed by using two primer sets that amplify selectively the corresponding hypervariable regions of the 16s region in bacteria (V2-V9). The Ion Torrent PGM platform was used for the performance of next-generation sequencing. RESULTS In the study samples, twelve phyla were identified. The most abundant ones were Firmicutes, Proteobacteria, Bacteroidetes, followed by Actinobacteria and Fusobacteria. Seventy-nine families, 79 genera and 137 species were identified and characterized. Moreover, 17 unique differentially abundant families, 30 unique differentially abundant genera and 24 unique differentially abundant species were identified among healthy subgroups with adjusted p-values <0.05. At the genus level, Moraxella (Proteobacteria) and Gemella (Firmicutes) were detected with a statistical significance in non-smokers, while Bifidobacterium (Actinobacteria), Alloscardovia (Actinobacteria), Dialister (Firmicutes) and Filifactor (Firmicutes) were present mostly in smokers. CONCLUSIONS The URT is colonized by a variety of protective and potentially pathogenic bacteria. This microbiome system is highly diverse and varies significantly between individuals. Geographic location and ethnicity are considered to be a strong determinants and factors affecting the diversity and abundance of the URT microbiome. Although some of the most abundant families are common irrespective of these factors, the dominance patterns are usually different between the study subjects and between the studies from other geographic locations. Unique differentially abundant families, genera and species were identified, and further studies are needed to elucidate their role. Further studies should focus on the investigation of the URT microbiome dynamics and the interaction with the host in health and disease. HIPPOKRATIA 2018, 22(1): 29-36.