S1 Fig. Rarefaction curves for all sequences at all sampling locations estimating the number
of bacterial OTUs at the 97% identity level.
S2 Fig. Stacked column bar graph representing the predicted metabolic attributes between
upstream and downstream sites against the KEGG database implemented in PICRUSt at
(A) tier level-1 and (B) tier level-2. The mean Nearest Sequenced Taxon Index (NSTI) value
for all samples was 0.188 ± 0.000125 s.d.
S3 Fig. Heat map and cluster analysis of bacterial core 97% identity OTUs between the different
sampling locations. Samples were grouped using hierarchical clustering (complete
linkage) based on the Bray–Curtis distance matrix calculated from the relative abundances (in
percent) of the OTUs. The colour code goes from blue (not detected) to yellow (low abundance)
to orange (medium abundance) to red (high abundance) on a logarithmic scale to
improve visualization between low and medium abundance.
S4 Fig. Network analysis of bacterial core OTUs (i.e. OTUs with relative abundance
>0.2%) and environmental parameters. Co-occurrence patterns were based on significant
(P<0.05) Spearman correlations with ρ ±0.6 showing the entire network structured according
to site (upstream, downstream and/or both). Each node (circle) in the network represents
a unique OTU and the size is proportional to node degree. Each edge (connection) represents
a strong and significant correlation (P<0.05), while the colour relates to the type of interaction:
positive (grey solid lines) or negative (red dashed lines). Environmental parameters are presented
by purple rectangles.
S1 Table. Summary of the sampling area, anthropogenic activities performed in the lower
WFS and associated contaminants.
S2 Table. Recommended Target Water Quality Range (TWQR) for the lower Wonderfonteinspruit.
The authors kindly thank IBIS/Universite´ Laval Plate-forme d’Analyses Ge´nomiques for 454-
pyrosequencing and support, especially Dr. Brian Boyle, as well as Marie-E`ve Beaulieu (manager
of DPK’s laboratory). We also acknowledge the Unit for Environmental Sciences and
Management (UESM), and David Hamman for assistance with sampling.