, 2011c). All these species have been involved in clinical infections and may bear several virulence genes, like those encoding Shiga toxins (stx1 and stx2), the type III secretion system

(TTSS) (ascF-G, Forskolin order ascV), flagella (fla) as well as several toxins (ast, act, alt, aexT) among others (Chacón et al., 2004; Aguilera-Arreola et al., 2005; Fehr et al., 2006; Chopra et al., 2009; Alperi & Figueras, 2010; Senderovich et al., 2012). Two new clinical species, Aeromonas taiwanensis and Aeromonas sanarellii, recovered from wound infections of hospitalized patients in Taiwan (although phenotypically misidentified as A. hydrophila and A. caviae, respectively) were recently discovered by sequencing the rpoD gene (Alperi et al., 2010a). Both species were described on the basis of a single strain (their type), and these were the only known Z-VAD-FMK purchase strains until two recent publications reported four isolates of A. sanarellii and one of A. taiwanensis in waste water in Portugal (Figueira et al., 2011), and a strain of A. taiwanensis recovered from the faeces of a female patient with diarrhoea in Israel (Senderovich et al., 2012). Isolates of the species A. sanarellii and A. taiwanensis were recorded in the course of a new study

that investigated the prevalence of Aeromonas populations in chironomid egg masses by culture and by real-time PCR methods (unpublished data). Considering the clinical relevance of these species, the Thalidomide present study describes for the first time the virulence genotypes and antibiotic susceptibility of these new species recovered from this new habitat and provides key phenotypic traits for their identification. Sampling for Aeromonas spp. populations was carried out in chironomid egg masses found in a waste stabilization pond in northern Israel between April and September 2009 using previously described procedures (Senderovich et al., 2008). Crushed egg masses were spread on M-Aeromonas agar (Biolife, Italy) for 24 h at 30 °C. Yellow, smooth, rounded colonies that were suspected Aeromonas species were then subcultured on Luria broth (LB) agar (Himedia, India). For each sample, about 15 Aeromonas isolates were identified to the species level using rpoD gene sequencing,

according to Soler et al. (2004). To observe the existence or not of clonally related isolates, DNA typing was carried out with the enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) technique using the primers and conditions described by Versalovic et al. (1991). Patterns with one or more different bands were considered different genotypes. In all A. sanarellii and A. taiwanensis strains, 24 phenotypic tests (Supporting information, Tables S1 and S2) were evaluated using conventional methods at 30 °C for 24 h up to 7 days as previously described (Abbott et al., 2003; Alperi et al., 2010b) with the exception of utilization of citrate, which was determined using the Simmons’s method (Cowan & Steel, 1993), and nitrate reduction (MacFaddin, 1976).