In contrast, only 15% (142/947) of library clones that were grouped into two OTUs (1 and 25) showed species-level sequence identity to Methanobrevibacter ruminantium, and only 4.3% (41/947) of clones populating two OTUs (11 and 14) displayed CP673451 nmr over 98% sequence identity to Methanobrevibacter smithii. Clones from 27 OTUs (21.1% or 200/947 of sequences from the combined libraries) only had 95-97.9% sequence identity to validly described Methanobrevibacter species (Tables
1 and 3), and likely corresponded to methanogen species that have yet to be cultivated. Based on 16S rRNA sequence identity, there is likely to be overlap between different hosts in representation of these uncharacterized methanogens, such as for instance AP5-146 (OTU 41) GSK2126458 clinical trial which was almost identical (1265/1268 bp) to the Ven09 methanogen clone identified in sheep from Venezuela [28]. Table 3 Percentage (%) in 16S rRNA gene clone distribution by taxon or phylum between alpacas Taxa Alpaca 4 Alpaca 5 Alpaca 6 Alpaca 8 Alpaca 9 Combined Methanobacteriales Methanobrevibacter ruminantium 1 16.2 11.6 7.0 28.6 12.3 15.0 millerae 1 57.5 32.7 62.7 27.5 57.0 47.3 smithii 1 6.1 5.0 3.5 4.8 2.2 4.3 unassigned 2 12.8 34.7 15.4 30.7 10.6
21.1 Methanobacterium unassigned 2 5.6 13.1 4.5 4.2 8.9 7.3 Methanosphaera unassigned 2 1.7 1.5 0.5 3.2 5.6 2.4 Thermoplasmatales unassigned 3 0.0 1.5 6.5 1.0 3.4 2.5 1sequences in OTUs that have 98% or higher sequence identity to the 16S rRNA gene of the specified species 2sequences in OTUs that have 95-97.9% sequence identity to the16S rRNA gene of a valid species from the specified genera 3sequences in OTUs that have 80-83% sequence identity to the 16S rRNA gene of Aciduliprofundum boonei and are likely part of a new order of uncultured archaea The remaining 14 OTUs were divided into three distinct phylogenetic groups. Clones from four OTUs (7, 19, 20 and 24), Selumetinib solubility dmso accounting for 7.3% (69/947) of the library sequences, showed 95-97.9% sequence identity to species
ID-8 belonging to the genus Methanobacterium (Table 3), and were accordingly grouped in the same cluster (Figure 2). Of interest in this category, clone AP4-007 from OTU 7 was almost identical (1259/1260 bp) to environmental clone UG3241.13 identified in dairy cattle from Canada [29]. Three other OTUs (13, 22 and 47), representing 2.4% (23/947) of clones, displayed genus-level sequence identity to Methanosphaera species and were also grouped into a single well-defined cluster by phylogenetic analysis (Figure 2). Finally, 2.5% (24/947) of alpaca clones were phylogenetically very distant from the previously mentioned genera within the order Methanobacteriales (Figure 2), and were grouped into 7 OTUs (15, 18, 28, 31, 35, 38 and 48) (Table 1).