However, all of the primer sets used
in these studies, which targeted three different variable regions of the 16S gene-the V4 region in the current study, V5 , and V6 regions [23, 24]-were shown in silico to cover the Bacteroidetes species, and the V4 primers were tested experimentally against genomic DNA from known Bacteroides isolates and shown to amplify 16 s rDNA. It is likely that members of the Bacteroidetes are also part of the core microbiome of porcine tonsils, despite the lack of evidence in our current data. While there were clear and strong similarities between the core microbiomes of all of the groups examined, there were also unique differences in minor genera found or missing from particular groups. https://www.selleckchem.com/products/elacridar-gf120918.html These differences can not readily be explained by differences in overall herd management or antibiotic p38 protein kinase usage in the groups (no antibiotics in Herd 1 time 1, Tylan in Herd 1 time 2, and Tylan plus Pulmotil in Herd 2). For example, reads identified as Arcanobacterium were found in all Herd 2 samples, and comprised 0.93% of the reads from that herd, but were not found in any Herd 1 sample. In contrast, reads identified
as Treponema were found in all but one sample from Herd 1, but not in any sample from Herd 2, and Chlamydia were found in Herd 1 tissue samples but not in Herd 2 samples. Lactobacillus was abundant in most samples from both Herd 1 time 1 and Herd 2, but was rare in Herd 1 time 2 samples. Pelosinus was abundant only in
Herd 1 time 1, not Herd 2 or Herd 1 time 2 samples. There were many other genera found in small numbers in 1-2 animals per group that were unique to that group, such as Polynucleobacter and Geobacter in Pig D from Herd 1 time 1 (Additional file 5), but no others that could be found in most animals in one group but not in animals of another group. These results indicate that, despite the small sample number, we can identify differences in the minor genera found in the two different herds. One goal of this project was to test tonsil brushes as an alternative, non-invasive method to collect tonsil samples, eliminating the need to euthanize animals to SB-3CT collect tonsil tissue. The Jaccard analysis (Figure 4) clearly indicated that all samples from the second sampling of Herd 1 were more similar to each other than to samples from Herd 1 and 2. We could detect differences between the brush and tissue extraction procedures as indicated in Figure 5, but the difference was small based on the range of eigenvalues. The detected statistical differences were a consequence of an increase in the percentage of reads identified as Actinobacillus, fewer sequences of Fusobacterium, Veillonella, and Peptostreptococcus), and no detectable sequences from the obligate intracellular pathogen Chlamydia in the brush specimens.