Pseudomonas putida 1 1 2 1 2 1 2 14 1 8 14 12 16 1997 1 21 12 5 Pseudomonas putida 8 27 imipenemipm IPM piperacillinceftazidimeamikacin norfloxacin 27 IPM IMP blaimp P. putida blaimp 27 8 9 4 1 MIC P. putida 1 9 blaimp Pseudomonas aeruginosa blaimp amikacin 4 gentamicin tobramycin P. putida P. aeruginosa Key words: Pseudomonas putidaimp Pseudomonas putida 5 I 1 1997 1 21 12 5 P. putida P. putida 2 MIC 1 imipenem IPM: meropenemmepm: ceftazidimecaz: cefepimecfpm: aztreonamazt: piperacillin PIPC: gentamicingm: tobramycintob: amikacinamk: norfloxacinnflx: SMA 2 PCR rifampicin RIF Pseudomonas aeruginosa PAO ML 517 CAZ8gmL RIF1gmL 4 PFGE Spe DNA BIO RAD P. aeruginosa 4 II 1 1997 1 21 12 5 P. putida 8 9 15
Pseudomonas putida Table 1 Isolation number and frequency of Pseudomonas putida strains Derived from Year 1997 1998 1999 2 21 Total Inpatients Outpatients 12 16 9 2 12 5 6981 14169 Total 15 16 12 2 17 8 Table 2 Souce of Pseudomonas putida isolates P. putida isolateno. Source urine respiratory digestive others total Imipenem resistant strains Imipenem susceptible strains 22 1 27 4 1 1 9 27 56 Total 2 27 14 1 8 8.169 Table 1 2 27 71 8 27 IPM 22 81 Table 2P. putida IPM 78 84 IPM Fig1 IPM AZT PIPCCAZAMKNFLX NCCLS P. putida 8 MIC MIC gml IMP16CAZ2 AZT2PIPC2GM16AMK16NFLX 16 2 27 IPM SMA PCR blaimp IMP PFGE MIC 27 8 1 9 4 1 P. putida PFGE Fig2 12 19 2 1182 29 4 22 27 2 17 118229 1 22 27 2 118 Isolation frequency 1 8 6 4 2 PIPC CAZ AZT GM AMK NFLX IPM-r IPM-s PIPC: piperacillin, CAZ: ceftazidime, AZT: aztreonam, GM: gentamicin, AMK: amikacin, NFLX: norfloxacin, IPM: imipenem Fig1. Isolation frequency of drug resistant strains in imipenemipm resistant or IPM susceptible Pseudomonas putida isolates. Fig2. Pulsed field gel electrophoresis patterns of Pseudomonas putida isolates derived from the same ward.
229 7 PFGE Table 1 MIC 1 AMK 1 GM TOB 8 NFLX PFGE MIC 118229 5 4 blaimp P. putida 1 9 P. aeruginosa CAZ P. putida 1 PCR blaimp P. aeruginosa MIC AMK GM TOB 4 blaimp Table 4 2 P. putida P. aeruginosa III 1991 P. aeruginosa 5 Serratia marcescens 67 Acinetobacter baumannii 8 Alcaligenes xylosoxidans 9 IMP Table Resistance patterns of strains derived from the same ward P. putida strain MICgmL Source strain no. isolation date IPM MEPM CAZ CFPM AZT PIPC GM TOB AMK NFLX 9 19988 urine 64 128 128 128 2 16 2 5 128 1 12 19996 urine 128 128 128 64 2 16 2 5 128 1 19 24 environment 64 128 128 128 2 16 2 5 64 1 1 1997 urine 2 128 128 64 2 2 64 2 2 128 18 24 environment 64 128 128 64 2 16 64 64 64 128 2 24 environment 64 128 128 64 2 2 128 128 128 128 29 218 urine 64 128 128 64 64 2 128 128 64 128 17 24 urine 128 128 128 128 64 64 128 64 128 128 22 27 urine 128 128 128 128 64 2 128 64 128 128 27 212 urine 128 128 128 128 64 2 128 64 128 128 16 2 urine 128 128 128 128 2 64 128 64 128 128 15 199912 urine 16 128 128 128 2 128 64 64 128 4 21 24 environment 64 128 128 128 2 16 64 2 128 1 IPM: imipenemmepm: meropenemcaz: ceftazidimecfpm: cefepimeazt: aztreonampipc: piperacillingm: gentamicin TOB: tobramycinamk: amikacinnflx: norfloxacin Table 4. Resistance patterns conferred by blaimp bearing plasmids Transconjugant MICgmL derived from IPM MEPM CAZ CFPM AZT PIPC GM TOB AMK No. 9 4 2 128 64 2 8.5.5 64 No. 12 4 2 128 128 2 4.5.5 64 No. 19 4 2 128 64 2 4.5.5 64 No. 29 4 2 128 64 2 4 4 16 128 No. 17 4 2 128 64 2 4.5.5 128 No. 22 4 2 128 64 2 4 4 16 2 No. 27 4 2 128 64 2 4 4 16 64 No. 15 4 2 128 64 2 4.5.5 128 No. 21 4 2 128 64 2 4 4 16 2 ML 517*.5.5.5 1 2 1.5.5 2 Host strain of transconjugants IPM: imipenemmepm: meropenemcaz: ceftazidimecfpm: cefepimeazt: aztreonampipc: piperacillingm: gentamicin TOB:tobramycinAMK: amikacin
Pseudomonas putida P. putida VIM 1 IMP 5 P. putida IPM IPM IMP IPM P. aeruginosa 6.5 11 A. baumannii 47.4 8 P. putida IPM Stenotrophomonas maltophila 12 Chryseobacterium spp. 1 P. putida BERGEYS MANUAL P. aeruginosa Pseudomonas fluorescens saprophytic or opportunistic animal pathogens 14 IPM IMP P. aeruginosa P. aeruginosa P. putida P. aeruginosa P. putida P. putida compromised host IMP 1 : MIC 1968 1974 Chemotherapy 29: 76791981 2 : 45: 8485 21 : 47: 1471511999 4 Yamashita MHida YSuzuki Set al.: Genetic Analysis of Pseudomonas aeruginosa by Pulsed Field Gel Electrophoresis 71: 67 611997 5 Watanabe MIyobe SInoue Met al.: Transferable imipenem resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother 5: 1471511991 6 Osano EArakawa YWacharotayankun Ret al.: Molecular characterization of an enterobacterial metallo lactamase found in a clinical isolate of Serratia marcescens that shows imipenem resistance. Antimicrob Agents Chemother 8: 7178 1994 7 Hirakata YIzumikawa KYamaguchi Tet al.: Rapid detection and evaluation of clinical characteristics of emerging multiple drug resistant gram negative rods carrying the metallo lactamase gene blaimp. Antimicrob Agents Chemother 42: 26 2111998 8 Takahashi AYomoda SIyobe S, et al.: Detection of carbapenemase producing Acinetobacter baumannii in a hospital. J. Clin. Microbiol. 8: 526529 2 9 Iyobe SKusadokoro HTakahashi Aet al.: Detection of a variant metallo LactamaseIMP 1from two unrelated strains of Pseudomonas aeruginosa and an Alcaligenes xylosoxidans strain. Antimicrob Agents Chemother 46: 214216 22 1 Lee KLim J BYum J Het al.: blavim 2 cassette containing novel integrons in metallo lactamase producing Pseudomonas aeruginosa and Pseudomonas putida isolates disseminated in a Korean hospital. Antimicrob Agents Chemother 46: 1515822 11 : 6 : 6 22 12 Saino YKobayashi FInoue Met al.: Purification and properties of inducible penicillin lactamase isolated from Pseudomonas maltophilia. Antimicrob Agents Chemother 22: 564571982 1 Bellais SAubert DNaas Tet al.: Molecular and biochemical heterogeneiety of class B carbapenem hydrolyzing lactamases in Chryseobacterium meningoseptcum. Antimicrob Agents Chemother 44: 187818862 14 Palleroni N J: Genus I. Pseudomonas Migula 1984. In Bergey s Manual of Systematic Bacteriologyvol 1Krieg N RHolt J Gedp. 141199Williams &WilkinsBaltimore1984
Isolation of carbapenem resistant Pseudomonas putida and its genetic background Sachie Yomoda 1 Ayako Takahashi 1 Toyoji Okubo 2 Masami Murakami 1 and Shizuko Iyobe 2 1 Department of Laboratory Medicine and Clinical Laboratory Center 2 Laboratory of Drug Resistance in Bacteria Gunma University School of Medicine 9 15 Showa machi, MaebashiJapan We isolated 8 Pseudomonas putida strains in the 5 years from January 1997 through December 21 at Gunma University Hospital. The sample was free of patient duplication. Among them27 isolates were resistant to imipenemipm22 of which were of urine origin. None was isolated from respiratory specimens. Most IPM resistant isolates were strains multiply resistant to piperacillinceftazidime amikacinand norfloxacin. The IMP metallo lactamase geneblaimpwas identified by PCR from all 27 IPM resistant strainswhich were derived from different 8 wards. We focused on 1 blaimp bearing P. putida strains of a ward9 isolatedfrominpatients and 4 detected from around the water pipe. The long term residence of blaimp bearing P. putida strainsidentified as the same strains with pulsed field gel electrophoresispfgepatterns and MIC patterns as for 1 drugswas observed in both inpatients and the ward environment. From 9 of the 1 strainsthe blaimp gene was effectively transferred to a recipient strain of Pseudomonas aeruginosaconferring resistance to IPM and other lactams concomitantly with amikacin resistance; 4 of the 9 strains conferred additional resistance to gentamicin and tobramycin.