Fig. 8

Co-occurrence of Legionella ASVs with protozoans Ciliophora, Amoebozoa, and Percolozoa. SPIEC-EASI cross-domain co-occurrence networks of Legionella ASVs and potential protozoan hosts, associated with high sodium concentrations, are presented for water and biofilm samples. Networks are based on next-generation sequencing (NGS) performed using Legionella spp.-specific 16S primers. Raw data were pre-filtered to retain Legionella amplicon sequence variants (ASVs). NGS amplicon sequencing targeting protozoa was performed using 18S rRNA primers directed to the V9 region. Raw data were pre-filtered to retain Amoebozoa, Ciliophora, and Percolozoa ASVs. We included ASVs present at a minimal frequency of 20 reads across all samples, in a minimum of 2 samples. To focus on Legionella spp. associated with high sodium concentrations, the analysis includes ASVs from Tiberias Hot Springs (THS, n = 20) and Haon borehole (n = 4) stations. Moreover, we only included Legionella ASVs present in the top 30 ASV importance table, of the random forest procedure (Fig. 4c,d). ASVs are represented by the last 5 letters of a letter-number code. For the full-length code and corresponding sequence, see Supplementary Data 1 and 2. Edges with low weights (> 0.01) and unconnected nodes were removed. Only positive associations between ASV nodes are presented. Edge width corresponds to association strength between ASVs