Fig. S1. Number of virulence genes found in each serotype in the isolates showing positive results for one or more genes. (A) All positive serotypes; (B) positive serotypes distributed by wastewater origin.

Table S1. Effector genes analysed in this study.

Table S2. Serotypes comprising the environmental collection. Serotyping of O and H antigens of the strains were determined according to the method described by Guinée and colleagues (1981) employing all available O (O1 to O181) and H (H1 to H56) antisera. All antisera were obtained and absorbed with the corresponding cross-reacting antigens to remove non-specific agglutinins.

Table S3. Oligonucleotides, target genes, binding sites and product size for detection of all genes analysed in this study. PCRs were performed with a GeneAmp PCR system 2400 (Perkin-Elmer, PE Applied Biosystems, Barcelona, Spain) and using the primers described in Table S3. The conditions for amplification were 1 cycle at 94°C and 5 min for denaturation, followed by 35 cycles at 94°C for 1.5 min, 1 min at the annealing temperature corresponding to each oligonucleotide and elongation at 72°C according to the product size (1 min for each 1000 bp). Finally, 1 cycle at 72°C for 4 min ended the amplification procedure. The PCR products were analysed by gel electrophoresis on a 1% agarose gel and bands were visualized by ethidium bromide staining. For sequencing the same primers were used.

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