RESUMO
We studied in parallel the population structure of 90 carbapenemase-producing and 88 carbapenemase-susceptible Klebsiella pneumoniae isolates collected in 20 Spanish hospitals, in the context of the EuSCAPE project. Fourteen and 50 multilocus sequence types (MLSTs) were detected among the carbapenemase-producing and carbapenem-susceptible isolates, respectively. ST11 and ST15 clones were more frequent in the carbapenemase-producing group than in the carbapenemase-susceptible group (P < 0.0001). Among the members of the carbapenem-suceptible group, the cefotaxime-resistant population showed population parameters that differed between the populations of the wild-type strains and the carbapenemase producers.
Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Carbapenêmicos/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , beta-Lactamases/metabolismo , Proteínas de Bactérias/genética , Cefotaxima/farmacologia , Farmacorresistência Bacteriana/genética , Klebsiella pneumoniae/enzimologia , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , beta-Lactamases/genéticaRESUMO
OBJECTIVES: There is little information about carbapenemase-producing (CP) Citrobacter spp. We studied the molecular epidemiology and microbiological features of CP Citrobacter spp. isolates collected in Spain (2013-15). METHODS: In total, 119 isolates suspected of being CP by the EUCAST screening cut-off values were analysed. Carbapenemases and ESBLs were characterized using PCR and sequencing. The genetic relationship among Citrobacter freundii isolates was studied by PFGE. RESULTS: Of the 119 isolates, 63 (52.9%) produced carbapenemases, of which 37 (58.7%) produced VIM-1, 20 (31.7%) produced OXA-48, 12 (19%) produced KPC-2, 2 (3.2%) produced NDM-1 and 1 (1.6%) produced VIM-2; 9 C. freundii isolates co-produced VIM-1 plus OXA-48. Fourteen isolates (22.2%) also carried ESBLs: 8 CTX-M-9 plus SHV-12, 2 CTX-M-9, 2 SHV-12 and 2 CTX-M-15. Fifty-seven isolates (90.5%) were C. freundii, 4 (6.3%) were Citrobacter koseri, 1 (1.6%) was Citrobacter amalonaticus and 1 (1.6%) was Citrobacter braakii. By EUCAST breakpoints, eight (12.7%) of the CP isolates were susceptible to the four carbapenems tested. In the 53 CP C. freundii analysed by PFGE, a total of 44 different band patterns were observed. Four PFGE clusters were identified: cluster 1 included eight isolates co-producing VIM-1 and OXA-48; blaVIM-1 was carried in a class 1 integron (intI-blaVIM-1-aacA4-dfrB1-aadA1-catB2-qacEΔ1/sul1) and blaOXA-48 was carried in a Tn1999.2 transposon. CONCLUSIONS: We observed the clonal and polyclonal spread of CP Citrobacter spp. across several Spanish geographical areas. Four species of Citrobacter spp. produced up to five carbapenemase types, including co-production of VIM-1 plus OXA-48. Some CP Citrobacter spp. isolates were susceptible to the four carbapenems tested, a finding with potential clinical implications.
Assuntos
Proteínas de Bactérias/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/enzimologia , Enterobacteriáceas Resistentes a Carbapenêmicos/genética , Citrobacter/enzimologia , Citrobacter/genética , Infecções por Enterobacteriaceae/microbiologia , Variação Genética , beta-Lactamases/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/classificação , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Citrobacter/classificação , Citrobacter/isolamento & purificação , Eletroforese em Gel de Campo Pulsado , Infecções por Enterobacteriaceae/epidemiologia , Genótipo , Humanos , Epidemiologia Molecular , Tipagem Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Espanha/epidemiologiaRESUMO
INTRODUCTION: An analysis was made about the evolution of resistance to 3rd generation cephalosporins, imipenem, and other antibiotics in invasive isolates of Klebsiella pneumoniae (K. pneumoniae) according to the Spanish EARS-Net database (2010-2014). METHODS: Forty-two hospitals from 16 Autonomous Communities with an approximate population coverage of 33% participated. RESULTS: A total 7,140 pneumoniae corresponding to the same number of patients were studied. Overall resistance percentages (I+R) were: cefotaxime 15.8%, ceftazidime 13.7%, imipenem 1.7%, ciprofloxacin 20.1%, tobramycin 14.1%, gentamicin 10.4%, and amikacin 1.9%. Resistance to 3rd generation cephalosporins increased from 9.8% (2010) to 19% (2014); to ciprofloxacin from 15.4% (2010) to 19.6% (2014); to gentamicin from 6.2% (2010) to 10.3% (2014) and to tobramycin from 7.1% (2010) to 14.2% (2014) (p<.001 in all cases). Combined resistance to 3rd generation cephalosporins, ciprofloxacin, and aminoglycosides increased from 3.3% (2010) to 9.7% (2014) (p<.001). Resistance to imipenem also increased from 0.27% (2010) to 3.46% (2014) (p<.001). A total of 121 isolates were resistant to imipenem, of which 104 (86%) produced carbapenemases: 74 OXA-48, 14 VIM, 9 KPC (6 KPC-2 and 3 KPC-3), 6 IMP, and 1 GES. CONCLUSIONS: Over the 5 year period (2010-2014), resistance to 3rd generation cephalosporins in invasive K. pneumoniae in Spain has doubled. The combined resistance to 3rd generation cephalosporins, ciprofloxacin, and aminoglycosides has tripled, and imipenem resistance has increased almost 13 times, mostly due to the spread of carbapenemase-producing isolates.
Assuntos
Antibacterianos/farmacologia , Bacteriemia/microbiologia , Cefalosporinas/farmacologia , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana Múltipla , Imipenem/farmacologia , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Aminoglicosídeos/farmacologia , Bacteriemia/epidemiologia , Proteínas de Bactérias/metabolismo , Área Programática de Saúde , Criança , Pré-Escolar , Ciprofloxacina/farmacologia , Infecção Hospitalar/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/enzimologia , Masculino , Pessoa de Meia-Idade , Vigilância da População , Espanha/epidemiologia , Adulto Jovem , Resistência beta-Lactâmica , beta-Lactamases/metabolismoRESUMO
OBJECTIVES: The objective of this study was to analyse the microbiological traits and the population structure of carbapenemase-producing (CP) Escherichia coli isolates collected in Spain between 2012 and 2014. METHODS: Two-hundred-and-thirty-nine E. coli isolates non-susceptible to carbapenems were studied. The carbapenemase genes and the phylogenetic groups were characterized using PCR. MLST was carried out using the typing schemes of the University of Warwick and the Institut Pasteur. The diversity of the population structure was estimated by calculating a simple diversity index (SDI). RESULTS: One-hundred-and-twenty-one isolates (50.6%) produced carbapenemases, of which 87 (71.9%) were OXA-48, 27 (22.3%) were VIM-1, 4 (3.3%) were KPC-2, 2 (1.7%) were NDM and 1 (0.8%) was IMP-22; 4 isolates were collected in 2012, 40 in 2013 and 77 in 2014. Ertapenem was more sensitive than imipenem or meropenem for screening for OXA-48-producing E. coli. Using the Warwick typing scheme, 59 different STs were identified, the most prevalent being ST131 (16.5%). The population diversity was higher among VIM-1-producing isolates (SDIâ=â81.5%) than among OXA-48-producing isolates (SDIâ=â44.8%). The Pasteur scheme had a higher discrimination capability (SDIâ=â55.4%) than the Warwick scheme (SDIâ=â48.8%). CONCLUSIONS: A progressive increase in the prevalence of CP E. coli was observed, mainly due to the dissemination of OXA-48 producers. The most sensitive method for detecting decreased susceptibility of CP E. coli to carbapenems was disc diffusion with ertapenem using the EUCAST screening cut-offs. The spread of CP E. coli was due to a polyclonal population. The Pasteur scheme showed the highest discrimination power. Surveillance is crucial for the early detection of CP E. coli.
Assuntos
Proteínas de Bactérias/metabolismo , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimologia , Variação Genética , Filogenia , beta-Lactamases/metabolismo , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Ertapenem , Escherichia coli/classificação , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Proteínas de Escherichia coli/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Tipagem de Sequências Multilocus , Reação em Cadeia da Polimerase , Prevalência , Espanha/epidemiologia , beta-Lactamases/genética , beta-Lactamas/farmacologiaRESUMO
OBJECTIVES: We analysed the microbiological traits and population structure of KPC-producing Enterobacteriaceae isolates collected in Spain between 2012 and 2014. We also performed a comparative WGS analysis of the three major KPC-producing Klebsiella pneumoniae clones detected. METHODS: Carbapenemase and ESBL genes were sequenced. The Institut Pasteur MLST scheme was used. WGS data were used to construct phylogenetic trees, to identify the determinants of resistance and to de novo assemble the genome of one representative isolate of each of the three major K. pneumoniae clones. RESULTS: Of the 2443 carbapenemase-producing Enterobacteriaceae isolates identified during the study period, 111 (4.5%) produced KPC. Of these, 81 (73.0%) were K. pneumoniae and 13 (11.7%) were Enterobacter cloacae. Three major epidemic clones of K. pneumoniae were identified: ST11/KPC-2, ST101/KPC-2 and ST512/KPC-3. ST11/KPC-2 differed from ST101/KPC-2 and ST512/KPC-3 by 27â819 and 6924 SNPs, respectively. ST101/KPC-2 differed from ST512/KPC-3 by 28â345 SNPs. Nine acquired resistance genes were found in ST11/KPC-2, 11 in ST512/KPC-3 and 13 in ST101/KPC-2. ST101/KPC-2 had the highest number of virulence genes (20). An 11 bp deletion at the end of the mgrB sequence was the cause of colistin resistance in ST512/KPC-3. CONCLUSIONS: KPC-producing Enterobacteriaceae are increasing in Spain. Most KPC-producing K. pneumoniae isolates belonged to only five clones: ST11 and ST512 caused interregional spread, ST101 caused regional spread and ST1961 and ST678 produced independent hospital outbreaks. ST101/KPC-2 had the highest number of resistance and virulence genes. ST101/KPC-2 and ST512/KPC-3 were recently implicated in the spread of KPC in Italy.
Assuntos
Infecções por Enterobacteriaceae/epidemiologia , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/enzimologia , Genoma Bacteriano , Genótipo , Análise de Sequência de DNA , beta-Lactamases/genética , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Enterobacteriaceae/classificação , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Humanos , Epidemiologia Molecular , Tipagem de Sequências Multilocus , Prevalência , Espanha/epidemiologiaRESUMO
INTRODUCTION: Clinical and epidemiological description of an outbreak in an intensive care unit (ICU) caused by a strain of multidrug-resistant Enterobacter cloacae complex carrying a CTX-M-9-type extended-spectrum ß-lactamase (ESBL). METHODS: A retrospective study of the clinical and epidemiological features of the outbreak caused by E.cloacae complex was performed. Identifying and studying the sensitivity of the strains were performed using the semi-automated system BD Phoenix™, and the characterisation of ESBL using PCR and sequencing. Molecular typing was performed by pulsed-field gel electrophoresis (PFGE). RESULTS: During February 2014, 6 patients (50% women; mean age: 61.5 years; age range: 44-76 years) admitted to the ICU of the Hospital of Pontevedra (CHOP) presented resistant E.cloacae complex isolates to extended-spectrum cephalosporins. Three patients developed infection; one had primary bacteraemia and septic shock, and 2 with ventilator-associated pneumonia. In the remaining three cases E.cloacae complex isolates were considered as colonisation. Phenotypic and genotypic analysis revealed that all isolates had the same PFGE profile and carried the same CTX-M-9 ESBL. The outbreak was controlled by improving universal precautions and contact isolation of patients infected and/or colonized. CONCLUSION: The clinical and epidemiological features of an outbreak in an ICU caused by E.cloacae complex carrying CTX-M-9 are described.
Assuntos
Surtos de Doenças , Farmacorresistência Bacteriana Múltipla , Enterobacter cloacae/efeitos dos fármacos , Infecções por Enterobacteriaceae/epidemiologia , Unidades de Terapia Intensiva , Adulto , Idoso , Eletroforese em Gel de Campo Pulsado , Enterobacter cloacae/enzimologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , beta-LactamasesRESUMO
We report the epidemiological impact of carbapenemase-producing Enterobacteriaceae (CPE) in Spain in 2012. Of the 237 carbapenemases detected, 163 were from the OXA-48 group, 60 were from VIM-1, 8 were from KPC-2, 5 were from IMP, and 1 was from NDM-1. Interhospital spread of carbapenemase-producing Klebsiella pneumoniae was due to a limited number of multilocus sequence types (MLST) and carbapenemase types, including ST15-VIM-1, ST11-OXA-48, ST405-OXA-48, ST101-KPC-2, and ST11-VIM-1. The number of CPE cases in Spain has increased sharply in recent years, due mainly to the emergence of OXA-48.
Assuntos
Proteínas de Bactérias/metabolismo , Enterobacteriaceae/enzimologia , beta-Lactamases/metabolismo , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , Humanos , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Espanha , beta-Lactamases/genéticaRESUMO
Little information is available about pediatric infections caused by extended-spectrum-ß-lactamase (ESBL)-producing Escherichia coli. We characterized an outbreak caused by a CTX-M-14-producing E. coli isolate in a neonatal intensive care unit (NICU) and studied other infections caused by ESBL-producing E. coli in non-NICU pediatric units. All children ≤4 years old who were infected or colonized by ESBL-producing E. coli isolates between January 2009 and September 2010 were included. Molecular epidemiology was studied by phylogroup analysis, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing. Antibiotic resistance genes were analyzed by PCR and sequencing. Plasmids were studied by PFGE with S1 nuclease digestion and by incompatibility group analysis using a PCR-based replicon-typing scheme. Of the ESBL-producing E. coli isolates colonizing or infecting the 30 newborns, identical PFGE results were observed for 21 (70%) isolates, which were classified as CTX-M-14-producing E. coli of ST23 phylogroup A. bla(CTX-M-14a) was linked to ISEcp1 and was carried on an â¼80-bp IncK plasmid. A smaller ongoing outbreak due to SHV-12-producing ST131 E. coli was also identified in the same NICU. Fifteen additional infections with ESBL-producing E. coli were identified in non-NICU pediatric units, but none was caused by the CTX-M-14-producing E. coli epidemic clone. Overall, CTX-M-14 (71.1%), CTX-M-15 (13.3%), and SHV-12 (13.3%) were the most important ESBLs causing pediatric infections in this study. Infections of newborns with CTX-M-14-producing E. coli were caused by both clonal and nonclonal isolates.
Assuntos
Antibacterianos/administração & dosagem , Surtos de Doenças , Infecções por Escherichia coli/tratamento farmacológico , Escherichia coli/efeitos dos fármacos , Criança , Pré-Escolar , Impressões Digitais de DNA , Eletroforese em Gel de Campo Pulsado , Escherichia coli/enzimologia , Escherichia coli/genética , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Feminino , Humanos , Lactente , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Masculino , Tipagem de Sequências Multilocus , Filogenia , Plasmídeos , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Espanha/epidemiologia , Inibidores de beta-Lactamases , beta-Lactamases/metabolismoRESUMO
We study the epidemiology, molecular basis, clinical risk factors, and outcome involved in the clonal dissemination of VIM-1-producing Klebsiella pneumoniae isolates in the hospital setting. All patients infected/colonized by carbapenem-nonsusceptible K. pneumoniae (CNSKP) in 2009 were included. Molecular epidemiology was studied by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Antibiotic resistance genes were analyzed by PCR and sequencing. Plasmids were studied by PFGE with S1 nuclease digestion and for incompatibility group by a PCR-based replicon typing scheme. Risk factors associated with CNSKP colonization/infection were assessed by an observational case-control study. All 55 patients studied were infected (n = 28) or colonized (n = 27) by VIM-1-producing K. pneumoniae. All but one acquired isolates of a single clone (PFGE cluster 1 [C1], sequence type 15 [ST15]), while another clone (PFGE C2, ST340) was detected in four patients. C1 isolates also produced the new extended-spectrum ß-lactamase SHV-134. bla(VIM-1) was carried in a class 1 integron and an untypeable plasmid of â¼50 bp. The number of days that the patient received mechanical ventilation, the use of parenteral nutrition, previous treatment with linezolid, and treatment with extended-spectrum cephalosporins for more than 7 days were detected to be independent risk factors for CNSKP acquisition. The VIM-1-producing K. pneumoniae ST15 clone has a high capacity to spread among intensive care unit patients with severe underlying conditions. A high rate of associated mortality and great difficulty in controlling the spread of this clone, without permanent behavioral changes in the personnel, were observed.
Assuntos
Antibacterianos/administração & dosagem , Carbapenêmicos/administração & dosagem , Infecção Hospitalar/tratamento farmacológico , Surtos de Doenças , Infecções por Klebsiella/tratamento farmacológico , Klebsiella pneumoniae/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Infecção Hospitalar/microbiologia , Infecção Hospitalar/mortalidade , Infecção Hospitalar/transmissão , Impressões Digitais de DNA , DNA Bacteriano/análise , DNA Bacteriano/biossíntese , Farmacorresistência Bacteriana Múltipla , Eletroforese em Gel de Campo Pulsado , Feminino , Hospitais , Humanos , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/mortalidade , Infecções por Klebsiella/transmissão , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus , Fatores de Risco , Espanha/epidemiologia , Taxa de Sobrevida , Resultado do Tratamento , beta-Lactamases/biossínteseRESUMO
Newborns are rarely infected by extended-spectrum ß-lactamase (ESBL)-producing members of the Enterobacteriaceae. In a neonatal intensive care unit, 14 newborns were infected or colonized by CTX-M-15-producing Enterobacter cloacae. All seven infected patients had underlying medical conditions, and five of them were treated successfully with meropenem, whilst one untreated patient died. Paediatric infections caused by multidrug-resistant ESBL-producing Enterobacter cloacae constitute a critical clinical and epidemiological issue.
Assuntos
Infecção Hospitalar/epidemiologia , Surtos de Doenças , Farmacorresistência Bacteriana Múltipla , Enterobacter cloacae/isolamento & purificação , Infecções por Enterobacteriaceae/epidemiologia , beta-Lactamases/metabolismo , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Análise por Conglomerados , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Eletroforese em Gel de Campo Pulsado , Enterobacter cloacae/efeitos dos fármacos , Enterobacter cloacae/enzimologia , Enterobacter cloacae/genética , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Enterobacteriaceae/microbiologia , Feminino , Genótipo , Humanos , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Masculino , Meropeném , Testes de Sensibilidade Microbiana , Tipagem Molecular , Tienamicinas/uso terapêutico , Resultado do TratamentoRESUMO
We describe a clinical case of an abdominal abscess due to NDM-1-producing Klebsiella pneumoniae in a 35-year-old Spanish patient after hospitalization in India for perforated appendicitis and peritonitis. The strain belonged to the MLST type 231 and had multiple additional antibiotic resistance genes such as bla(CTX-M-15), armA methylase, aac(6')-Ib-cr, dfrA12, sul1 and qnrB and lack of porin genes ompK35 and ompK36. The patient was cured after abscess drainage.