Using the “”Phylogenetic Analysis”" tool within MG-RAST, the GS20

Using the “”Phylogenetic Analysis”" tool within MG-RAST, the GS20 and FLX sequencing runs were searched against the RDP and greengenes databases using the BLASTn algorithm. The percent of sequences assigned to each

of the bacterial phyla from the pig fecal GS20 (A and B) and FLX (C and D) metagenomes #BIX 1294 in vivo randurls[1|1|,|CHEM1|]# is shown. The e-value cutoff for 16S rRNA gene hits to RDP and greengenes databases was 1×10-5 with a minimum alignment length of 50 bp. Both GS20 and FLX metagenomic swine fecal datasets were dominated by Firmicutes and Bacteroidetes phyla (Figure 1), which is consistent with several molecular phylogenetic studies of mammalian gut environments, including the swine gut [2, 8, 10, 14]. Archaeal sequences constituted less than 1% of total rRNA gene sequences retrieved in either swine metagenome, and were dominated by the Methanomicrobia and Thermococci, which is consistent with previous molecular diversity studies of pig manure [16]. While

these populations are only a very small fraction of the total microbiota [17], methanogens contribute significantly to the metabolic potential within in a gut environment [18]. The majority of eukaryotic sequences derived from the swine metagenomes are related to Chordata (i.e., host phylum), fungi, and the Viridiplantae (i.e., feed). Sequences sharing high sequence homology to Balantidium coli were obtained in both swine metagenomes. The latter is many a protozoan pathogen that causes balantadiasis in mammalian hosts, including human and swine. Since the samples were collected from healthy animals, these CX-5461 in vivo sequences might be associated with non-pathogenic B. coli strains or with pathogenic strains in asymptomatic animals. Viral sequences were rare, comprising less than 1% of the total metagenomic sequences when compared to the SEED database (Additional File 1, Fig. S1). The low abundance of viral sequences retrieved from the swine fecal metagenomes is consistent with viral proportions retrieved in termite, chicken, and cattle gastrointestinal metagenomes, and may be a direct result of limited

representation of viral genetic information in currently available databases [8]. A closer look at the taxonomic distribution of the numerically abundant bacterial orders derived from the swine metagenomes revealed that Clostridiales, unclassified Firmicutes, Bacteroidales, Spirochaetales, unclassified gammaproteobacteria, and Lactobacillales were the top six most abundant bacterial groups (Additional File 1, Fig. S2). At the genus-level taxonomic resolution, Prevotella species were the most abundant, comprising 19-22% of 16S rRNA gene sequences within both swine fecal metagenomes (Additional File 1, Fig. S3). Of the classified Clostridiales, Sporobacter was the next most abundant genus within both the swine fecal metagenomic datasets.

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