| Abstract | Objectives To analyse the recombination events associated with conjugal mobilization of two multiresistance plasmids, pRUM17i48 and pLAG (formerly named pDO1-like), from Enterococcus faecium 17i48 to Enterococcus faecalis JH2-2. Methods The plasmids from two E. faecalis transconjugants (JH-4T, tetracycline resistant, and JH-8E, erythromycin resistant) and from the E. faecium donor (also carrying a pHTβ-like conjugative plasmid, named pHTβ17i48) were investigated by several methods, including PCR mapping and sequencing, S1-PFGE followed by Southern blotting and hybridization, and WGS. Results Two locations of repApHTβ were detected in both transconjugants, one on a â^¼50âEUR?kb plasmid (as in the donor) and the other on plasmids of larger sizes. In JH-4T, WGS disclosed an 88.6âEUR?kb plasmid resulting from the recombination of pHTβ17i48 (â^¼50âEUR?kb) and a new plasmid, named pLAG (35.3âEUR?kb), carrying the tet(M), tet(L), lsa(E), lnu(B), spw and aadE resistance genes. In JH-8E, a 75âEUR?kb plasmid resulting from the recombination of pHTβ17i48 and pRUM17i48 was observed. In both cases, the cointegrates were apparently derived from replicative transposition of an IS1216 present in each of the multiresistance plasmids into pHTβ17i48. The cointegrates could resolve to yield the multiresistance plasmids and a pHTβ17i48 derivative carrying an IS1216 (unlike the pHTβ17i48 of the donor). Conclusions Our results completed the characterization of the multiresistance plasmids carried by the E. faecium 17i48, confirming the role of pHT plasmids in the mobilization of non-conjugative antibiotic resistance elements among enterococci. Results also revealed that mobilization to E. faecalis was associated with the generation of cointegrate plasmids promoted by IS1216-mediated transposition. |
| Text | 382544 2017 10.1093/jac/dkx197 Scopus 2 s2.0 85035132650 ISI Web of Science WOS 000408084700007 Plasmids Enterococcus faecalis Enterococcus faecium pHT promoted mobilization of non conjugative resistance plasmids from Enterococcus faecium to Enterococcus faecalis Di Sante L.; Morroni G.; Brenciani A.; Vignaroli C.; Antonelli A.; D Andrea M.M.; Cesare A.D.; Giovanetti E.; Varaldo P.E.; Rossolini G.M.; Biavasco F. Unit of Microbiology, Department of Life and Environmental Sciences, Polytechnic University of Marche, Ancona, , Italy; Unit of Microbiology, Department of Biomedical Sciences and Public Health, Polytechnic University of Marche Medical School, Ancona, , Italy; Department of Experimental and Clinical Medicine, University of Florence, Florence, , Italy; Department of Medical Biotechnologies, University of Siena, Siena, , Italy; Microbial Ecology Group, CNR Institute of Ecosystem Study, Verbania, , Italy; Microbiology and Virology Unit, Florence Careggi University Hospital, Florence, , Italy Objectives To analyse the recombination events associated with conjugal mobilization of two multiresistance plasmids, pRUM17i48 and pLAG formerly named pDO1 like , from Enterococcus faecium 17i48 to Enterococcus faecalis JH2 2. Methods The plasmids from two E. faecalis transconjugants JH 4T, tetracycline resistant, and JH 8E, erythromycin resistant and from the E. faecium donor also carrying a pHTβ like conjugative plasmid, named pHTβ17i48 were investigated by several methods, including PCR mapping and sequencing, S1 PFGE followed by Southern blotting and hybridization, and WGS. Results Two locations of repApHTβ were detected in both transconjugants, one on a a ¼50aEUR kb plasmid as in the donor and the other on plasmids of larger sizes. In JH 4T, WGS disclosed an 88.6aEUR kb plasmid resulting from the recombination of pHTβ17i48 a ¼50aEUR kb and a new plasmid, named pLAG 35.3aEUR kb , carrying the tet M , tet L , lsa E , lnu B , spw and aadE resistance genes. In JH 8E, a 75aEUR kb plasmid resulting from the recombination of pHTβ17i48 and pRUM17i48 was observed. In both cases, the cointegrates were apparently derived from replicative transposition of an IS1216 present in each of the multiresistance plasmids into pHTβ17i48. The cointegrates could resolve to yield the multiresistance plasmids and a pHTβ17i48 derivative carrying an IS1216 unlike the pHTβ17i48 of the donor . Conclusions Our results completed the characterization of the multiresistance plasmids carried by the E. faecium 17i48, confirming the role of pHT plasmids in the mobilization of non conjugative antibiotic resistance elements among enterococci. Results also revealed that mobilization to E. faecalis was associated with the generation of cointegrate plasmids promoted by IS1216 mediated transposition. 72 Published version http //www.scopus.com/inward/record.url eid=2 s2.0 85035132650 partnerID=q2rCbXpz Articolo 2017_J_Antimicrob_Chemother_2017_72_2447_2453.pdf Articolo in rivista Academic Press for the British Society for Antimicrobial Chemotherapy, 0305 7453 Journal of Antimicrobial Chemotherapy. Print Journal of antimicrobial chemotherapy Print Journal of antimicrobial chemotherapy Print J. antimicrob. chemother. Print andrea.dicesare DI CESARE ANDREA |
