The 0.03 OTU curves were different with that of the unique OTU (Fig. 1B). The most marked change happened to A, B and D groups, which three showed dissimilar slopes this time. The condition D showed the steepest slope, suggesting that more tags in the group having larger than 3% variance than the other two conditions. The difference between E and B curves for 0.03 OTU was less pronounced than that for the unique OTU, indicating that a proportion of different unique sequences between B and E groups were within 97% similarity, which could possibly be produced by the PCR mutation. In addition to unique and 0.03 OTUs, we also compared OTUs at 0.05 and 0.10 distances (Additional file
2), and the trends were generally similar to that for 0.03 OTU. Nevertheless, because the larger distance OTUs harbored more varied sequences, the differences between the 5 groups were less obvious. Abundance of top 300 tags The Fig. #Selleck DZNeP randurls[1|1|,|CHEM1|]# 2 presents the relative abundance of the top 300 V6 sequences in the 10 samples. We observed that the E group (blue curve) showed significant differences with the other four groups, particularly for many tags within the top 50 abundances. For instance, the 10th abundant tag assigned as Syntrophobacterales (Deltaproteobacteria) showed 0.95-1.19% abundance in A to D groups, but only occupied 0.03-0.06% in the E group. The 15th abundant tag assigned as Epsilonproteobacteria had abundances of 0.46-0.62% in group A to D samples, but showed
1.50-1.53% in the learn more MRIP E group. In total, 91 out of the top 300 tags in group E showed significant differences with other 8 samples using the students t-test analysis (p < 0.01). A further PCA analysis using the 300 tags proved that the E1 and E2 were obviously different with other 8 samples (Fig. 2). Figure 2 Relative abundances (%) of the top 300 predominant V6 sequences in the 10 samples. The right figure shows the PCA of the 10 samples using the abundance data of top 300 tags. Microbial community
structure The community structure was compared at the phylum (subphylum for proteobacteria) level (Fig. 3). In general, the A to D groups showed very similar structure, but the E group showed obvious differences. The A-D groups showed higher phylum evenness than the E group. Statistically, the E group had higher percentage of Gammaproteobacteria and Epsilonproteobacteria, but lower percentage of Chloroflexi and Planctomycetes (One Way ANOVA, p < 0.01). We also compared the 10 samples using clustering with Primer 6 (Fig. 3). The result showed that samples E1 and E2 formed a different branch with the other 8 samples. Figure 3 Relative abundance of bacteria phyla (subphyla) in the 10 samples. The dendrogram shows the clustering of 10 samples using the phyla (subphyla) abundance data. Discussion Sequencing quality The present study sequenced the 16 S rRNA V6 tags using the Solexa platform, which employed a different base calling procedure with the pyrosequencing .