Costas C Papagiannitsis1, Vincenzo Di Pilato2, Tommaso Giani3, Panagiota Giakkoupi4, Eleonora Riccobono5, Giulia Landini3, Vivi Miriagou6, Alkiviadis C Vatopoulos7, Gian Maria Rossolini8. 1. Department of Microbiology, National School of Public Health, Athens, Greece Department of Microbiology, Faculty of Medicine and University Hospital in Plzen, Charles University in Prague, Plzen, Czech Republic. 2. Department of Medical Biotechnologies, University of Siena, Siena, Italy Department of Surgery and Translational Medicine, University of Florence, Florence, Italy. 3. Department of Medical Biotechnologies, University of Siena, Siena, Italy. 4. Department of Microbiology, National School of Public Health, Athens, Greece. 5. Department of Medical Biotechnologies, University of Siena, Siena, Italy Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy. 6. Laboratory of Bacteriology, Hellenic Pasteur Institute, Athens, Greece. 7. Department of Microbiology, National School of Public Health, Athens, Greece Central Public Health Laboratory, Hellenic Centre of Disease Control and Prevention, Vari, Greece. 8. Department of Medical Biotechnologies, University of Siena, Siena, Italy Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy Microbiology and Virology Unit, Careggi University Hospital, Florence, Italy Don Carlo Gnocchi Foundation, Florence, Italy gianmaria.rossolini@unifi.it.
Abstract
OBJECTIVES: Global dissemination of KPC-type carbapenemases is mainly associated with the spread of high-risk clones of Klebsiella pneumoniae and of KPC-encoding plasmids. In this study, we explored the population structure of KPC-encoding plasmids from the recent epidemics of KPC-producing K. pneumoniae (KPC-Kp) in Greece and Italy, the two major European endemic settings. METHODS: Thirty-four non-replicate clinical strains of KPC-Kp representative of the early phases (2008-11) of the Greek (n = 22) and Italian (n = 12) epidemics were studied. Isolates were typed by MLST, and blaKPC-carrying plasmids were characterized by S1 profiling, PCR-based replicon typing and RFLP. Transfer experiments by conjugation or transformation were carried out with Escherichia coli recipients. Eleven plasmids, representative of all different restriction profiles, were completely sequenced. RESULTS: The representative Greek strains belonged to 14 sequence types (STs), with a predominance of ST258. The representative Italian strains belonged to three STs, with a predominance of clonal complex 258 (ST258, ST512). The 34 strains carried plasmids of variable size (78-166 kb), either with blaKPC-2 or blaKPC-3 gene embedded in a Tn4401a transposon. Plasmids from Greek strains were mostly of a single RFLP type (A) and resembled the archetypal pKpQIL KPC-encoding plasmid, while plasmids from Italian strains belonged to a more heterogeneous population, showing five RFLP profiles (A, C-F). Types A and C resembled pKpQIL or deletion derivatives thereof, while types D-F included plasmids with hybrid structures between pKpQIL, pKPN3 and pKPN101-IT. CONCLUSIONS: pKpQIL-like plasmids played a major role in the dissemination of blaKPC in Greece and Italy, but evolved with different dynamics in these endemic settings.
OBJECTIVES: Global dissemination of KPC-type carbapenemases is mainly associated with the spread of high-risk clones of Klebsiella pneumoniae and of KPC-encoding plasmids. In this study, we explored the population structure of KPC-encoding plasmids from the recent epidemics of KPC-producing K. pneumoniae (KPC-Kp) in Greece and Italy, the two major European endemic settings. METHODS: Thirty-four non-replicate clinical strains of KPC-Kp representative of the early phases (2008-11) of the Greek (n = 22) and Italian (n = 12) epidemics were studied. Isolates were typed by MLST, and blaKPC-carrying plasmids were characterized by S1 profiling, PCR-based replicon typing and RFLP. Transfer experiments by conjugation or transformation were carried out with Escherichia coli recipients. Eleven plasmids, representative of all different restriction profiles, were completely sequenced. RESULTS: The representative Greek strains belonged to 14 sequence types (STs), with a predominance of ST258. The representative Italian strains belonged to three STs, with a predominance of clonal complex 258 (ST258, ST512). The 34 strains carried plasmids of variable size (78-166 kb), either with blaKPC-2 or blaKPC-3 gene embedded in a Tn4401a transposon. Plasmids from Greek strains were mostly of a single RFLP type (A) and resembled the archetypal pKpQIL KPC-encoding plasmid, while plasmids from Italian strains belonged to a more heterogeneous population, showing five RFLP profiles (A, C-F). Types A and C resembled pKpQIL or deletion derivatives thereof, while types D-F included plasmids with hybrid structures between pKpQIL, pKPN3 and pKPN101-IT. CONCLUSIONS: pKpQIL-like plasmids played a major role in the dissemination of blaKPC in Greece and Italy, but evolved with different dynamics in these endemic settings.
Authors: Marco Coppi; Vincenzo Di Pilato; Francesco Monaco; Tommaso Giani; Pier Giulio Conaldi; Gian Maria Rossolini Journal: Antimicrob Agents Chemother Date: 2020-03-24 Impact factor: 5.191
Authors: Michelle M C Buckner; Howard T H Saw; Rachael N Osagie; Alan McNally; Vito Ricci; Matthew E Wand; Neil Woodford; Alasdair Ivens; Mark A Webber; Laura J V Piddock Journal: mBio Date: 2018-04-24 Impact factor: 7.867
Authors: Sophia David; Victoria Cohen; Sandra Reuter; Anna E Sheppard; Tommaso Giani; Julian Parkhill; Gian Maria Rossolini; Edward J Feil; Hajo Grundmann; David M Aanensen Journal: Proc Natl Acad Sci U S A Date: 2020-09-23 Impact factor: 11.205