SUPPLEMENTARY FIG. S1. (Continued).

Transcription

1 Supplementary Data SUPPLEMENTARY FIG. S1. Plasmid profile study by PFGE-S1-Nuclease. The gels show the approximate number and size of the large plasmids present in each isolate. Isolates were numbered consecutively above each lane in the same order as they appear in Supplementary Table 1. (A) Escherichia coli harbored one to five plasmids that ranged < kb in size. (B) Klebsiella oxytoca harbored one to six plasmids that ranged < kb in size. (C) Citrobacter freundii harbored one to six plasmids that ranged < kb in size. (D) Serratia marcescens harbored one to three plasmids that ranged < kb in size. *NS, sample not studied; PFGE, pulsed-field gel electrophoresis.

2 SUPPLEMENTARY FIG. S1. (Continued).

3 SUPPLEMENTARY FIG. S2. Number of isolates and hospitals referring KPC-producing non-klebsiella pneumoniae isolates from June 2010 to December The graph shows the number of KPC-producing ETB (squares), the number of hospitals referring isolates (triangles), and the cumulative number of hospitals (diamonds). The study included isolates that were referred until March This year was not included in this graph because it was misleading. ETB, Enterobacteriaceae; KPC, Klebsiella pneumoniae carbapenemase.

4 Specie ID a Supplementary Table S1. Epidemiological and Molecular Characteristics of KPC-Producing Non-K. pneumoniae Enterobacteriaceae HTAL. code Province Date Sex Age Sample PFGE type ESBLs b bla KPC genetic environment c No. of plasmids/mw (Kb) ECO-1 NEU NEUQUEN June-2010 Female 55 Pleural fluid B No.4 4/<48.5; 60; 76; 145 ECO-2 HHE NEUQUEN January-2011 Male 87 Blood G c CTX-M-1/15 No.2 2/<48.5; 135 ECO-3 FAV BA CITY August-2010 ND 45 Rectal screening C No.8 2/<48.5; 83 ECO-4 BAR RIO NEGRO September-2010 Female 19 Rectal screening D c Tn4401a 2/66; 145 ECO-5 JAF BA CITY December-2010 Male 43 Rectal screening F Tn4401a 2/<48.5; 104 ECO-6 SWI BA CITY October-2010 Female 94 Rectal screening E Tn4401a 2/<48.5; 66; 120 ECO-7 HRA CORDOBA March-2011 Male 61 Urine H No.2 1/83 ECO-8 HOC BA CITY May-2011 Male 87 Urine I No.8 2/83; 102 ECO-9 VIR BA CITY May-2011 Female ND Tracheal aspirate J c CTX-M-1/15 Tn4401a 2/91; 135 ECO-10 FAV BA CITY May-2011 Female 44 Urine K No.8 3/83; 86; 140 ECO-11 PEN BA CITY April-2011 Male ND Rectal screening A Tn4401a 2/101; 135 ECO-12 PEN BA CITY June-2011 Female ND Rectal screening A Tn4401a 2/101; 135 ECO-13 HIM CORDOBA June-2011 Female 17 Purulent collection of ileostomy L No.3 1/75 ECO-14 CSF BA CITY September-2011 Female ND Rectal screening M No.14 2/<48.5; 145 ECO-15 HOA BA October-2011 Male 71 Mini bronchoalveolar N c CTX-M-1/15 Tn4401a 1/201 lavage fluid ECO-16 ANC BA CITY February-2012 Female 60 Rectal screening O Tn4401a 2/55; 97 ECO-17 CMM BA April-2012 ND ND Abdominal drainage NT CTX-M-8/25 Tn4401a nd ECO-18 HDB BA August-2012 Male 45 Abdominal drainage P Tn4401a 3/85; 104; 121 ECO-19 HJP CHACO September-2012 Male ND Abdominal drainage Q No.2 2/60; 91 ECO-20 COS BA CITY November-2012 Male ND Rectal screening R CTX-M-9/14 Tn4401a 4/52; 83; 104; 158 ECO-21 ITA BA CITY March-2013 Female 84 Feces S Tn4401a 3/<48.5; 55; 80 ECO-22 ALV BA CITY April-2013 Male ND Rectal screening T CTX-M-8/25 Tn4401a 5/<<48.5; <48.5; 66; 145; 199 ECO-23 ITA BA CITY July-2013 Male 79 Feces U CTX-M-1/15 No.8 3/83; 91; 194 ECO-24 HOB BA CITY August-2013 ND ND Abdominal drainage V Tn4401a 3/62; 130; 255 ECO-25 MIT BA CITY September-2013 Male 32 Bone W Tn4401a 2/80; 178 ECO-26 GUT BA CITY October-2013 Male 5 Retroculture X No.2 3/<48.5; 91; 154 ECO-27 JAF BA CITY January-2014 Male 50 ND Y No.8 3/<48.5; 66; 158 ECO-28 ITA BA CITY March-2014 Male 90 Urine J c CTX-M-1/15 Tn4401a 1/102 ECO-29 DUR BA CITY June-2014 ND 56 Abdominal drainage H No.8 1/83 KOX-1 EVI BA August-2011 Female 5 m Blood E No.2 2/90; 196 KOX-2 POS BA September-2011 Male 9 Retroculture G No.6 4/<48.5; 145; 160; 437 KOX-3 EVI BA November-2012 Male 58 Lymph node biopsy F No.2 3/59; 135; 185 KOX-4 VIR BA CITY Dec-2010 Female 84 Blood K CTX-M-2 No.2 4/<48.5; 145; 129; 310 KOX-5 IFL BA CITY April-2011 Male 82 Rectal screening L PER-2 No.2 3/80; 182; 214 KOX-6 ITA BA CITY August-2011 Male 25 Abdominal drainage A No.2 4/48.5; 59; 129; 176 KOX-7 GUT BA CITY October-2012 Male 3 m Urine B No. 3/150; 199; 299 KOX-8 COS BA CITY November-2012 Male 38 Blood C PER-2 No.2 2/170; 310 KOX-9 GUT BA CITY November-2012 Female 17 m Urine R No.2 2/<48.5; 188 KOX-10 GUT BA CITY October-2013 Female 4 Retroculture G No.6 6/<48.5; 59; 150; 185; 349; 360 KOX-11 GUT BA CITY November-2013 Male 5 cc M PER-2 No.2 3/60; 129; 340 KOX-12 JAF BA CITY Jan-2014 ND ND Dialisis water N PER-2 No.2 4/<48.5; <48.5; 150; 340 KOX-13 ITA BA CITY February-2014 Male 2 m Articular joint fluid O No.10 2/60; 145 KOX-14 HSR CORDOBA October-2010 ND ND Retroculture D No.2 3/59; 145; 373 KOX-15 HIM CORDOBA January-2011 Male 7 Blood D No.2 2/145; 340 KOX-16 HIM CORDOBA July-2011 Male 1 Blood D No.2 3/59; 145; 340 KOX-17 HMU CORDOBA May-2012 Male 52 Skin and soft tissue H PER-2 No.2 4/59; 110; 145; 209 KOX-18 JP2 CORRIENTES December-2013 Male 5 Blood P No.2 4/<48.5; 49; 86; 196 KOX-19 HJP CHACO November-2012 Female ND Urine J PER-2 No.2 4/<48.5; 59; 114; 310 KOX-20 H09 MENDOZA May-2011 ND ND ND A CTX-M-2 No.2 5/48.5; 59; 129; 145; 176 KOX-21 HAC RIO NEGRO October-2013 Female 43 Blood Q PER-2 No.2 3/<48.5; 90; 340 KOX-22 LPT TUCUMAN December-2010 Male ND Urine I No.2 1/237 CFR-1 EVI BA December-2012 Male 72 Drainage G No.2 3/59; 97; 261 CFR-2 ALV BA CITY November-2010 Male ND Urine E No.2 3/52; 62; 108 CFR-3 HVS BA CITY April-2012 Male ND Skin and soft tissue M No.2 1/48.5 CFR-4 ITAL BA CITY October-2012 Male 73 Blood F No. 2/57; 69 CFR-5 GUT BA CITY November-2012 Female ND ND D No. 2/291; 368 CFR-6 GUT BA CITY December-2013 Female ND ND D No. 2/97; 284 CFR-7 ALV BA CITY June-2013 Female ND Urine E PER-2 No. 2/66; 322 CFR-8 ALV BA CITY June-2013 Male 72 Retroculture E No.13 2/66; 291 CFR-9 ALV BA CITY June-2013 Male ND Urine E PER-2 No.13 2/66; 322 CFR-10 GUT BA CITY October-2012 Male 12 Rectal screening D No.5 3/97; 252; 274 CFR-11 GUT BA CITY April-2013 Male 3 Rectal screening D No.2 3/104; 252; 284 CFR-12 HSP CHACO August-2010 Female 50 Skin and soft tissue L No.2 2/48.5; 170 CFR-13 HJP CHACO June-2013 Male ND Skin and soft tissue L No.2 2/<48.5; 194 CFR-14 HSR CORDOBA June-2011 Female 37 Urine H No. 4/<48.5; 48.5; 68; 97; 136; 174 CFR-15 HSR CORDOBA November-2011 Male 61 Urine H No. 5/64; 91; 130; 165 CFR-16 NSM CORDOBA January-2012 Male 56 Blood I No.2 5/64; 91; 130; 267 CFR-17 HMU CORDOBA May-2012 Male 52 Skin and soft tissue N PER-2 No.2 2/55; 173 CFR-18 HPS JUJUY April-2013 Male 49 Urine O CTX-M-2 No. 4/48.5; 58; 97; 123 CFR-19 H09 MENDOZA September-2010 ND ND ND J No.2 5/<48.5; 60; 97; 145; 300 (continued)

5 Specie ID a Supplementary Table S1. (Continued) HTAL. code Province Date Sex Age Sample PFGE type bla KPC genetic environment c No. of plasmids/mw (Kb) ESBLs b CFR-20 NJT TUCUMAN August-2012 Male 3 m Catheter K CTX-M-1/15, PER-2 SMA-1 PBA BA CITY June-2010 ND ND Urine E No. 2/<48.5; SMA-2 UOM BA CITY April-2012 Male 43 Abdominal drainage C No.2 2/48.5; 79.5 SMA-3 UOM BA CITY September-2012 Male 31 Abdominal drainage C CTX-M-2 No.2 3/48.5; 79.5; 254 SMA-4 FLE BA CITY September-2012 Female ND Blood F No.7 1/79.5 SMA-5 COS BA CITY Jan-2013 Female ND Blood G No.2 2/<48.5; 48.5 SMA-6 COS BA January-2013 Male ND ND G No.2 1/48.5 No.9 5/48.5; 100; 145; 215; 300 SMA-7 PIR BA CITY November-2013 Male 17 Mini bronchoalveolar lavage fluid D Tn4401a 1/79.5 SMA-8 SAS BA CITY January-2014 Male 41 Cerebrospinal fluid K No.2 1/<48 SMA-9 CEN BA September-2010 Female 20 Blood H No.2 2/<48.5; 70 SMA-10 SML BA April-2013 Male 65 Tracheal aspirate I Tn4401a 1/57 SMA-11 H09 MENDOZA February-2013 ND ND Blood A CTX-M-9/14 No.2 2/<48; SMA-12 H09 MENDOZA February-2013 ND ND Blood A CTX-M-9/14 No.12 2/<48.5; 296 SMA-13 H09 MENDOZA February-2013 ND ND Blood A CTX-M-9/14 No.12 2/<48.5; 296 SMA-14 H09 MENDOZA February-2013 ND ND Blood A CTX-M-9/14 No. 3/<48.5; 70; 296 SMA-15 HRA CORDOBA May-2011 Male ND Blood B No.2 2/70; 333 SMA-16 HRA CORDOBA September-2012 Female 38 Urine B No.11 3/70; 333; 369 SMA-17 HRA CORDOBA September-2012 Female 58 Blood B No.2 2/70; 333 SMA-18 VSW CORDOBA September-2012 Male 46 Abdominal abscess J PER-2 No.2 2/<48.5; 48.5 a The isolates were listed and therefore numbered consecutively as they appear in Supplementary Fig. S1 (PFGE-S1 nuclease assay). b ESBLs were screened by polymerase chain reaction as explained in Materials and Methods section. c Isolates confirmed as Escherichia coli ST131. HTAL., hospital; Date, month, and year in which the isolate was obtained; BA, Buenos Aires; BA City, Buenos Aires City; PFGE, pulsedfield gel electrophoresis; ND, not determined; nt, non-typeable.

6 Supplementary Table S2. List of Primers Used in the Study Primer name Nucleotide sequence Refs. CTX-MU1 ATGTGCAGYACCAGTAARGT Pagani et al. 1 CTX-MU2 TGGGTRAARTARGTSACCAGA Pagani et al. 1 CTX-F GCCGCTCAATGTTAACGGTGA Melano et al. 2 CTX-R ACCGTGGGTTACGATTTTCGC Melano et al. 2 CTXM8/25G-F CTGGAGAAAAGCAGCGGGGG This work CTXM8/25G-R CGCTGCCGGTTTTATCCCCGAC This work CTXM9G-F ATGGTGACAAAGAGAGTGCAACG This work CTXM9G-R GCGGCTGGGTAAAATAGGTCACC This work CTXM1/15G-F CAGTTCACGCTGATGGCGACG This work CTXM1/15G-R CGGCGCACGATCTTTTGGCCA This work PER-1-578F GGCCTGACGATCTGGAACCTT This work PER-6F GCCCTGATGATCTGGAGCCTT This work PER-U1-8R TAACCGCTsTGGTCCTGTGGT This work PER-2-PLUS GTA GTA TCA GCC CAA TCC CC Melano et al. 2 PER-2-MINUS CCA ATA AAG GCC GTC CAT CA Melano et al. 2 rfb.1bis mplx ATACCGACGACGCCGATCTG Blanco, et al. 3 rfbo25b.r mplx TGCTATTCATTATGCGCAGC Blanco, et al. 3 1 Pagani, L., E. Dell Amico, R. Migliavacca, M.M. D Andrea, E. Giacobone, G. Amicosante, E. Romero, and G.M. Rossolini Multiple CTX-M-type extended-spectrum b-lactamases in nosocomial isolates of Enterobacteriaceae from a hospital in northern Italy. J. Clin. Microbiol. 41: Melano, R., A. Corso, A. Petroni, D. Centrón, B. Orman, A. Pereyra, N. Moreno, and M. Galas Multiple antibiotic-resistance mechanisms including a novel combination of extended-spectrum beta-lactamases in a Klebsiella pneumoniae clinical strain isolated in Argentina. J Antimicrob. Chemother. 52: Blanco, M., M.P. Alonso, M.H. Nicolas-Chanoine, G. Dahbi, A. Mora, J.E. Blanco, C. López, P. Cortés, M. Llagostera, V. Leflon- Guibout, B. Puentes, R. Mamani, A. Herrera, M.A. Coira, F. García-Garrote, J.M. Pita, and J. Blanco Molecular epidemiology of Escherichia coli producing extended-spectrum b-lactamases in Lugo (Spain): dissemination of clone O25b:H4-ST131 producing CTX-M- 15. J. Antimicrob. Chemother. 63: