A molecular microbial ecology approach to studying hemodialysis water and fluid  M. Gomila, J. Gascó, J. Gil, R. Bernabeu, V. Iñigo, J. Lalucat  Kidney International  Volume 70, Issue 9, Pages 1567-1576 (November 2006) DOI: 10.1038/sj.ki.5001756 Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 1 Schematic representation of the study. Kidney International 2006 70, 1567-1576DOI: (10.1038/sj.ki.5001756) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 2 Sampling. Four samples were collected: point 1 after the first water treatment system, which performs chlorination, particle filtering, decalcification, and reverse osmosis of community water, before it enters the reservoirs; point 2, at the outlet of the repositories collecting pretreated water; point 3, at the connection port of the distribution ring, before water enters the dialysis monitor; and point 4, was the dialysis fluid at the dialyzer, after the dialysis bath was reconstituted and passed through a polysulfone filter. Samples were taken 1 day before, and 2 days after routine monthly hyperchlorination (30p.p.m.) disinfection, with sodium hypochlorite. The hypochlorite circulated through the distribution loop during the disinfection process during 12h at room temperature. All samples were collected disinfecting the sample port with ethanol at 70%. The sample 4 was collected directly by disconnecting the dialysate line from the dialyzer. Sampling points will be numbered 1, 2, 3, or 4, and samples classified as (BH) before or (AH) after the hyperchlorination step. Kidney International 2006 70, 1567-1576DOI: (10.1038/sj.ki.5001756) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 3 Phylogenetic affiliation of the sequenced clones in the Alphaproteobacteria group. Clones are designated by sampling point (2, 3, or 4), before or after hyperchlorination (B or A), CL followed by the clone number. 16S sequences of isolates obtained from the same sampling, reported in a previous study,3 are also included in the phylogenetic trees (strains designation indicated by MG). Accession numbers are indicated in brackets. Kidney International 2006 70, 1567-1576DOI: (10.1038/sj.ki.5001756) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 4 Phylogenetic affiliation of the sequenced clones in the Betaproteobacteria group. Clones are designated by sampling point (2, 3, or 4), before or after hyperchlorination (B or A), CL followed by the clone number. 16S sequences of isolates obtained from the same sampling, reported in a previous study,3 are also included in the phylogenetic trees (strains designation indicated by MG). Accession numbers are indicated in brackets. Kidney International 2006 70, 1567-1576DOI: (10.1038/sj.ki.5001756) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 5 Richness curves of the clones calculated by rarefaction. (a) Phylotype richness at sampling points 2, 3, and 4 before hyperchlorination (BH), and point 3 after hyperchlorination (AH). (b) Phylotypes and species richness, with consideration of the diversity in the entire system. Kidney International 2006 70, 1567-1576DOI: (10.1038/sj.ki.5001756) Copyright © 2006 International Society of Nephrology Terms and Conditions