The internal review boards and ethics committees of all collaborating hospitals

in the surveillance network approved the protocol, and written informed consent was collected from the guardians of all participants to obtain fecal and/or blood samples, and selleck compound use the clinical and microbiologic information for scientific studies [1]. The ST213 strain YU39 was used as a pA/C donor, since this was the only strain capable of conjugal transfer [5]. This strain harbored five plasmids: the 150 kb pA/C and four plasmids of different sizes (ca. 100, 40, 5 and 3 kb), for which no information was available. We selected strain SOHS 02-2 (hereafter referred to as SO1) which contains a 94 kb pSTV and a cryptic 80 kb plasmid [4], and the reference strain LT2 which only carries the 94 kb pSTV [8], as representative strains of the ST19 genotype harboring pSTV. The pSTV of SO1 and LT2 were marked with a kanamycin resistance cassette inserted into the spvC gene (coding for a phosphothreonine lyase) according to the Datsenko and Wanner protocol [9]. These strains were named SO1pSTV::Km

and LT2pSTV::Km, and were used as recipients in conjugation experiments (Table 1). Table 1 Bacterial strains and plasmids used in this work Strain Plasmids (kb) Feature Salmonella     YU39 (ST213) pA/C (150), p100 (100), pX1 PARP inhibitor (40), pColE1-like (5), p3 (3) Donor SO1 (ST19) pSTV::Km (94), p80 (80) Recipient LT2 (ST19) pSTV::Km (94) Recipient E. coli     DH5α   Recipient HB101   Recipient HB101pSTV pSTV::Km Ureohydrolase Recipient DH5α pA/C Wild-type pA/C, donor DH5α pA/C, pSTV::Km Stability assays DH5α pX1 Wild-type pX1 Transconjugants     SO1     IA4 pA/C Re-arranged pA/C IA5 pA/C Re-arranged pA/C IA9 pA/C Re-arranged pA/C IIA4 pA/C + pX1 pA/C and pX1 FRAX597 co-integrate HB101     IC2 pX1::CMY pX1 with

the transposed CMY region IIC1 pX1::CMY pX1 with the transposed CMY region IIIC9 pA/C + pX1 pA/C and pX1 co-integrate IIIC10 pX1::CMY pX1 with the transposed CMY region IVC8 pA/C + pX1 pA/C and pX1 co-integrate HB101pSTV ::Km     ID1 pX1::CMY pX1 with the transposed CMY region IID2 pX1::CMY pX1 with the transposed CMY region IIID8 pA/C + pX1 pA/C and pX1 co-integrate IVD2 pA/C + pX1 pA/C and pX1 co-integrate IVD8 pX1::CMY pX1 with the transposed CMY region LT2     IIE2 pX1::CMY pX1 with the transposed CMY region IIIE4 pX1::CMY pX1 with the transposed CMY region IIIE9 pA/C + pX1 pA/C and pX1 co-integrate DH5α     221-1 pA/C + pX1 pA/C and pX1 co-integrate 221-10 pA/C + pX1 pA/C and pX1 co-integrate 225-1 pA/C + pX1 pA/C and pX1 co-integrate 225-7 pA/C + pX1 pA/C and pX1 co-integrate pX1 mutants     DH5α pX1ydgA::Tn5 Tn5 transposon insertion DH5α pX1taxB::Km taxB site-directed mutant DH5α pA/C, pX1ydgA::Tn5 Donor DH5α pA/C,pX1taxB::Km Donor Transformation of pA/C and pSTV into E.