Statistical Analysis 16S & WGS datasets: Alpha & Beta diversity (PcoA), Differential Abundance & PERMANOVA
Various bacterial diversity metrics were employed for inferring the structural aspects of the microbial community. The alpha diversity was calculated to determine by Shannon index & Fisher’s alpha, Chao richness and evenness (Figure 1A). The results of both16s amplicon and shot gun metagenome demonstrated similar diversity patterns at height H2 (Figure 1B). Other comparisons indicate major diversity shifts at all heights. The rarefraction curve of all samples calculated had reached a plateau, suggesting that the sequencing was deep enough at phylum and genus levels (Figure 1C). Good’s coverage estimation revealed 80.3% to 91.0% species in all the samples.
Supervised PCA and CCA results (Figure 2), demonstrate separate clusters (significant differential distribution of fecal microbiota at p = 0.05), between altitudes H1 & H2 employing both 16s rRNA (Figure 2 Ai and Aii) and WGS methods (Figure 2 Bi, Bii). Global microbial community composition using Permutational Analysis of Multivariate Dispersions (PERMDISP) 2 clustering, also indicated significant differential distribution between H1 and H2 heights (Figure2Aiii&Biii), as well as all four heights H1, H2, H3 and H4 (Figure 2 C & D).