RESUMO
AIM: To investigate molecular epidemiology of extended-spectrum ß-lactamase/ESBL and plasmid-mediated AmpC ß-lactamase/pAmpC producing Gram-negative bacteria causing urinary tract infections (UTIs) in Zenica-Doboj Canton, Bosnia and Herzegovina, in the period Decembar 2009-May 2010. METHODS: Antibiotic susceptibility was determined by disc diffusion and broth microdilution according to CLSI guidelines. Double-disk synergy test was performed in order to screen for ESBLs/pAmpC beta-lactamases. PCR was used to detect bla(ESBL)/bla(ampC)/bla(carb) genes. Genetic relatedness of the strains was determined by pulsed-field-gel-electrophoresis (PFGE). RESULTS: Among 85 patients with UTIs caused by ESBL producing isolates, 44 (51.8%) were from in-patients and 41 (48.2%) from outpatients. Klebsiella spp. was the most frequently isolated from in-patients, in 28 (63.6%) cases. Among outpatients, Klebsiella spp./Escherichia coli were the most frequently isolated, in 19 (46.3%)/16 (39.0%) cases. Twenty-one (75.0%) from hospital and nine (47.4%) from outpatient Klebsiella spp. isolates were positive for blaTEM, whereas 27 (96.4%) from in-patients and 6 (31.6%) from outpatient were bla(CTX-M) positive (18 hospital and five outpatient isolates were encoding bla(CTX-M-15)). One Klebsiella oxytoca and one Enterobacter cloacae inpatient isolates were positive for blaCTX-M-28. One Klebsiella pneumoniae outpatient isolate were positive for bla(CTX-M-22) and one E. coli for bla(CTX-M-3). One hospital Proteus mirabilis strain was positive for bla(CMY-2) and two Klebsiella spp. strains for blaDHA-1, whereas two E. coli, one K. oxytoca and one Proteus vulgaris outpatient strains were positive for bla(CMY-2). CONCLUSION: Identification of bla(CTX-M-3), bla(CTX-M-22) and bla(CTX-M-28) among Enterobacteriaceae is uncommon. In this study we report the emergency of CMY-2 and DHA-1 plasmid-mediated beta-lactamases.
Assuntos
Enterobacteriaceae/enzimologia , Infecções Urinárias/epidemiologia , Infecções Urinárias/microbiologia , Assistência Ambulatorial , Proteínas de Bactérias/genética , Bósnia e Herzegóvina/epidemiologia , Citrobacter freundii/enzimologia , Citrobacter freundii/genética , Citrobacter freundii/isolamento & purificação , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Eletroforese em Gel de Campo Pulsado , Enterobacter cloacae/enzimologia , Enterobacter cloacae/genética , Enterobacter cloacae/isolamento & purificação , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Escherichia coli/enzimologia , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Proteínas de Escherichia coli/genética , Hospitalização , Humanos , Lactente , Klebsiella oxytoca/enzimologia , Klebsiella oxytoca/genética , Klebsiella oxytoca/isolamento & purificação , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/isolamento & purificação , Pessoa de Meia-Idade , Epidemiologia Molecular , Tipagem Molecular , Plasmídeos/genética , Prevalência , Proteus mirabilis/enzimologia , Proteus mirabilis/genética , Proteus mirabilis/isolamento & purificação , Infecções Urinárias/genética , beta-Lactamases/genéticaRESUMO
Salmonella enterica displaying resistance to extended-spectrum cephalosporins and fluoroquinolone (FQs) has been deemed a high-priority pathogen by the World Health Organization (WHO). While CTX-M type acquired ß-lactamases have been detected in S. enterica serovar Bovismorbificans, DHA enzymes have been rarely reported in S. Bovismorbificans. In this study, we here report for the first time the isolation of two multi-drug resistant (MDR) S. Bovismorbificans strains co-harboring plasmid-encoded AmpC (pAmpC) ß-lactamase gene (blaDHA-1) and qnrB gene, 16Sal017 isolated from a chicken meat sample and 16Sal018 from a grass carp fish sample, collected from retail markets in Guangzhou, China. The blaDHA-1 and qnrB genes in these two strains were both located on the same novel 217,773 bp IncHI2 plasmid belonged to ST2. The plasmid contained 16 additional acquired antimicrobial resistance genes encoding resistance to eight antibiotic classes and quaternary ammonium compound. Besides, 16Sal017 contained an additional 10,124 bp Col (pHAD28)-like plasmid harboring qnrS1. The blaDHA-1 and qnrB4 genes were located in an 18,198 bp region, sul1-qacEΔ1-ampR-blaDHA-1-pspABCDF-qnrB4-sapABC-IS91-sul1-qacEΔ1, which has been identified in various bacteria species, indicating the high transfer ability of blaDHA-1 and qnrB4 genes within this gene cluster. The IncHI2 plasmid was found to be transferable to Escherichia coli J53 by conjugation and resulted in the acquiring of multiple resistance in the transconjugants. Genome sequence comparisons by cgMLST and MAUVE alignment indicated 16Sal017 and 16Sal018 are highly similar and are not epidemiologically linked with strains from other sources and countries. Our findings suggest S. Bovismorbificans as a new host for conjugative mega-plasmid harboring blaDHA-1 and qnrB4 genes, and highlight the potential transmission opportunity of these S. Bovismorbificans clones through the food chain, which need continuous investigation.