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1.
Mikrobiyol Bul ; 55(1): 1-16, 2021 Jan.
Artigo em Turco | MEDLINE | ID: mdl-33590977

RESUMO

Antibiotic resistance is one of the most important public health problem and one of the most critical steps in preventing resistance is the monitorization of the resistance. Local, regional and global monitoring enables the spread of antibiotic resistance to be understood more clearly. In this study, it was aimed to evaluate the results of the pilot study for the establishment of molecular-based carbapenem surveillance system in Escherichia coli and Klebsiella pneumoniae isolates and to investigate the carbapenemase epidemiology in Turkey. Hospitals (n= 28) from 26 different statistical level II regions from Turkey were included in the study. The hospitals participated in the study submitted ten carbapenem susceptible and ten carbapenem resistant E.coli and K.pneumoniae isolates to our laboratory that were isolated in two different periiods of six-month either between 1 March-31 August or 1 April-30 September 2019. A total of 509 isolates were collected from 26 of the 28 participating hospitals in the study. Isolates were identified by matrix assisted laser desorptionization-time of flight mass spectrophotometry (MALDI TOF MS) (Bruker Daltonics, Germany) method and antibiotic susceptibility tests for imipenem, meropenem and colistin were studied by broth microdilution. Moreover, susceptibilities to amikacin, amoxicillin-clavulanic acid, ampicillin, aztreonam, cefepime, cefotaxime, ceftazidime, ciprofloxacin, ertapenem, gentamicin, piperacillin-tazobactam, tobramycin and trimethoprim-sulfamethoxazole were determined by disc diffusion method. The resistance genes were investigated in isolates which were found to be phenotypically resistant to carbapenem and colistin, in house method was used to investigate carbapenemase genes and a commercial colistin resistant real-time PCR kit (Biospeedy, Turkey) was used for colistin resistance genes. In total, 493 of the 509 isolates collected from hospitals were identified as E.coli (25.7%, n= 127) and K.pneumoniae (74.3%, n= 366) and included in the study. It was determined that 31% of the isolates evaluated were from community-acquired infections and 69% were either from healthcare-associated infections or from colonization sites. Among the tested isolates, 248 (50.3%) were susceptible to carbapenems and 245 (49.7%) were resistant. The types of carbapenemases in carbapenemase-producing were OXA-48 (52.2%), KPC (16.1%), NDM-1 (15%), OXA-48 + NDM-1 (12.6%), KPC + NDM-1 (2.8%) and VIM (0.5%) and OXA-48+VIM (0.5%). Resistance to colistin was detected in 23.3% of the isolates but mcr1-8 genes were not detected. It was found that all colistin resistant isolates are resistant to at least one of the carbapenems. The importance of a molecular-based antimicrobial resistance surveillance system in our country was demonstrated with this pilot study. It is thought that continuous monitoring of these epidemiological features will contribute to the management of infections due to carbapenemase-producing organisms.


Assuntos
Proteínas de Bactérias , Infecções por Klebsiella , Klebsiella pneumoniae , beta-Lactamases , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Escherichia coli/enzimologia , Escherichia coli/genética , Humanos , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana , Projetos Piloto , Turquia/epidemiologia , beta-Lactamases/genética
3.
Epidemiol Infect ; 149: e12, 2020 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-33327984

RESUMO

The prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae urinary tract infections (UTIs) is increasing worldwide. We investigated the prevalence, clinical findings, impact and risk factors of ESBL E. coli/K. pneumoniae UTI through a retrospective review of the medical records of children with UTI aged <15 years admitted to Prince of Songkla University Hospital, Thailand over 10 years (2004-2013). Thirty-seven boys and 46 girls had ESBL-positive isolates in 102 UTI episodes, compared with 85 boys and 103 girls with non-ESBL isolates in 222 UTI episodes. The age of presentation and gender were not significantly different between the two groups. The prevalence of ESBL rose between 2004 and 2008 before plateauing at around 30-40% per year, with a significant difference between first and recurrent UTI episodes of 27.3% and 46.5%, respectively (P = 0.003). Fever prior to UTI diagnosis was found in 78.4% of episodes in the non-ESBL group and 61.8% of episodes in the ESBL group (P = 0.003). Multivariate analysis indicated that children without fever (odds ratio (OR) 2.14, 95% confidence interval (CI) 1.23-3.74) and those with recurrent UTI (OR 2.67, 95% CI 1.37-5.19) were more likely to yield ESBL on culture. Congenital anomalies of the kidney and urinary tract were not linked to the presence of ESBL UTI. In conclusion, ESBL producers represented one-third of E. coli/K. pneumoniae UTI episodes but neither clinical condition nor imaging studies were predictive of ESBL infections. Recurrent UTI was the sole independent risk factor identified.


Assuntos
Infecções por Escherichia coli/microbiologia , Escherichia coli/efeitos dos fármacos , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Infecções Urinárias/microbiologia , beta-Lactamases/metabolismo , Adolescente , Antibacterianos/farmacologia , Criança , Pré-Escolar , Farmacorresistência Bacteriana Múltipla , Escherichia coli/enzimologia , Feminino , Humanos , Lactente , Klebsiella pneumoniae/enzimologia , Masculino , Estudos Retrospectivos , beta-Lactamases/genética
4.
Artigo em Inglês | MEDLINE | ID: mdl-33375538

RESUMO

The emergence and spread of antimicrobial resistance pose a threat to public health globally. Antibiotic-resistant bacteria and genes can disseminate among environments, animals and humans. Therefore, investigation into potential reservoirs of multidrug-resistant bacteria is of great importance to the understanding of putative transmission routes of resistant bacteria and resistance genes. This study aimed to report the occurrence of Escherichia coli harboring the Klebsiella pneumoniae carbapenemase-producing gene (blaKPC) in Psittaciformes rescued from wildlife trafficking in Paraíba State, Brazil. Cloacal swabs were collected from thirty birds and cultured by conventional microbiology using MacConkey and serum tryptone glucose glycerol (STGG) media supplemented with selective antimicrobials. E. coli isolates (n = 43) were identified by phenotypic tests and confirmed by MALDI-TOF. Antimicrobial susceptibility profiles were determined by means of Kirby-Bauer test. All isolates were further screened for extended-spectrum beta-lactamase (ESBL) production, and putative genes encoding ESBL were investigated by PCR. Additionally, blaKPC-harboring strains were genotyped by REP-PCR. A total of 43 E. coli phenotypically resistant isolates were recovered. The highest resistance rate was observed against ciprofloxacin. Among the resistance genes, only blaKPC was found in seven different birds from three species. According to the genotyping, these seven isolates belonged to four different strains. To date, this is the first report on the occurrence of KPC-E. coli in Psittaciformes rescued from trafficking in Northeastern Brazil. Due to the high clinical importance of KPC-E. coli, our findings suggest that wild animals in captivity at wildlife rescue centers can play a role as reservoirs of bacteria that are resistance to Critically Important antimicrobials in human medicine.


Assuntos
Proteínas de Bactérias/genética , Escherichia coli , Psittaciformes/microbiologia , beta-Lactamases/genética , Animais , Antibacterianos/farmacologia , Brasil/epidemiologia , Crime , Farmacorresistência Bacteriana Múltipla , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana
5.
S Afr Med J ; 110(8): 783-790, 2020 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-32880307

RESUMO

BACKGROUND: Rates of healthcare-associated infections (HAIs) among babies born in developing countries are higher than among those born in resource-rich countries, as a result of suboptimal infection prevention and control (IPC) practices. Following two reported deaths of neonates with carbapenem-resistant Klebsiella pneumoniae bloodstream infections (BSIs), we conducted an outbreak investigation in a neonatal unit of a regional hospital in Gauteng Province, South Africa. OBJECTIVES: To confirm an outbreak of K. pneumoniae BSIs and assess the IPC programme in the neonatal unit. METHODS: We calculated total and organism-specific BSI incidence risks for culture-confirmed cases in the neonatal unit for baseline and outbreak periods. We conducted a clinical record review for a subset of cases with K. pneumoniae BSI that had been reported to the investigating team by the neonatal unit. An IPC audit was performed in different areas of the neonatal unit. We confirmed species identification and antimicrobial susceptibility, and used polymerase chain reaction for confirmation of carbapenemase genes and pulsed-field gel electrophoresis (PFGE) for typing of submitted clinical isolates. RESULTS: From January 2017 to August 2018, 5 262 blood cultures were submitted, of which 11% (560/5 262) were positive. Of 560 positive blood cultures, 52% (n=292) were positive for pathogenic organisms associated with healthcare-associated BSIs. K. pneumoniae comprised the largest proportion of these cases (32%; 93/292). The total incidence risk of healthcare-associated BSI for the baseline period (January 2017 - March 2018) was 6.8 cases per 100 admissions, and that for the outbreak period (April - September 2018) was 10.1 cases per 100 admissions. The incidence risk of K. pneumoniae BSI for the baseline period was 1.6 cases per 100 admissions, compared with 5.0 cases per 100 admissions during the outbreak period. Average bed occupancy for the entire period was 118% (range 101 - 133%), that for the baseline period was 117%, and that for the outbreak period was 121%. In a subset of 12 neonates with K. pneumoniae bacteraemia, the median (interquartile range (IQR)) gestational age at birth was 27 (26 - 29) weeks, and the median (IQR) birth weight was 1 100 (880 - 1 425) g. Twelve bloodstream and 31 colonising K. pneumoniae isolates were OXA-48-positive. All isolates were genetically related by PFGE analysis (89% similarity). Inadequate IPC practices were noted, including suboptimal adherence to aseptic technique and hand hygiene (57% overall score in the neonatal intensive care unit), with poor monitoring and reporting of antimicrobial use (pharmacy score 55%). CONCLUSIONS: Overcrowding and inadequate IPC and antimicrobial stewardship contributed to a large outbreak of BSIs caused by genetically related carbapenemase-producing K. pneumoniae isolates in the neonatal unit.


Assuntos
Bacteriemia/microbiologia , Infecção Hospitalar/microbiologia , Surtos de Doenças , Unidades Hospitalares , Infecções por Klebsiella/epidemiologia , Gestão de Antimicrobianos , Bacteriemia/epidemiologia , Proteínas de Bactérias/metabolismo , Auditoria Clínica , Infecção Hospitalar/epidemiologia , Aglomeração , Humanos , Incidência , Recém-Nascido , Controle de Infecções , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Programas Médicos Regionais , África do Sul/epidemiologia , beta-Lactamases/metabolismo
6.
PLoS One ; 15(8): e0237474, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32857767

RESUMO

The effective treatment of carbapenemase-producing Klebsiella pneumoniae infection has been limited and required novel potential agents. Due to the novel drug development crisis, using old antimicrobial agents and combination therapy have been highlighted. This study focused on fosfomycin which inhibits cell wall synthesis and has potential activity on Enterobacteriaceae. We evaluated fosfomycin activity against carbapenemase-producing K. pneumoniae and characterized fosfomycin resistance mechanisms. Fosfomycin revealed effective activity against only 31.8% of carbapenemase-producing K. pneumoniae isolates. The major resistance mechanism was FosA3 production. The co-occurrence of FosA3 overexpression with the mutation of glpT (or loss of glpT) and/or uhpT was mediated high-level resistance (MIC>256 mg/L) to fosfomycin. Moreover, fosA3 silenced in sixteen fosfomycin-susceptible isolates and the plasmid carrying fosA3 of these isolates increased 32- to 64-fold of fosfomycin MICs in Escherichia coli DH5α transformants. The in vitro activity of fosfomycin combination with amikacin by checkerboard assay showed synergism and no interaction in six (16.2%) and sixteen isolates (43.3%), respectively. No antagonism of fosfomycin and amikacin was observed. Notably, the silence of aac (6)'-Ib and aphA6 was observed in amikacin-susceptible isolates. Our study suggests that the combination of fosfomycin and amikacin may be insufficient for the treatment of carbapenemase-producing K. pneumoniae isolates.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Farmacorresistência Bacteriana/genética , Fosfomicina/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , beta-Lactamases/metabolismo , Amicacina/farmacologia , Substituição de Aminoácidos , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/genética , Escherichia coli/metabolismo , Humanos , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/patologia , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Testes de Sensibilidade Microbiana , Plasmídeos/genética , Plasmídeos/metabolismo , RNA Mensageiro/metabolismo , beta-Lactamases/genética
7.
Rev Soc Bras Med Trop ; 53: e20190526, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32578705

RESUMO

INTRODUCTION: This study investigated the genetic environment of bla KPC-2 in Klebsiella pnemoniae multi-drug resistant clinical isolates. METHODS: Four carbapenemase gene isolates resistant to carbapenems, collected from infected patients from two hospitals in Brazil, were investigated using polymerase chain reaction and plasmid DNA sequencing. RESULTS: The bla KPC-2 gene was located between ISKpn6 and a resolvase tnpR in the non-Tn4401 element (NTEKPC-IId). It was detected on a plasmid belonging to the IncQ1 group. CONCLUSIONS: To our knowledge, this is the first report of the presence of the bla KPC-2 gene in the NTEKPC-IId element carried by plasmid IncQ1 from infections in Brazil.


Assuntos
Antibacterianos/farmacologia , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , beta-Lactamases/genética , DNA Bacteriano/genética , Farmacorresistência Bacteriana Múltipla , Humanos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Testes de Sensibilidade Microbiana , Plasmídeos/genética , Reação em Cadeia da Polimerase
8.
BMC Infect Dis ; 20(1): 416, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32539687

RESUMO

BACKGROUND: Klebsiella pneumoniae (KP) is the primary pathogen associated with pyogenic liver abscesses (PLAs). Moreover, there has been an increase in the proportion of extended-spectrum beta-lactamase (ESBL)-producing KP. However, the clinical and computed tomography (CT) features of liver abscesses caused by ESBL-producing KP have not been separately described. We aimed to compare the clinical and CT features present in patients with ESBL-producing and non-ESBL-producing KP as well as to determine the risk factors for ESBL-producing KP liver abscesses (KPLAs). METHODS: We performed a retrospective analysis of data obtained from the medical records of patients with a first episode of KPLA admitted to Shengjing Hospital of China Medical University between May 2015 and May 2019. We compared the clinical and CT features between patients with ESBL-producing and non-ESBL-producing KPLA. RESULTS: We enrolled 100 patients with KPLA (14 and 86 in the ESBL-producing and non-ESBL-producing groups, respectively). There was no significant between-group difference in the proportion of patients with comorbid diabetes (71.43% vs. 66.2%, p = 0.086). The ESBL-producing KPLA group had a greater proportion of patients with a history of biliary disease (78.57% vs. 26.74%, p < 0.001) and gastrointestinal malignancy (50% vs. 6.98%, p < 0.001). Multivariate regression analysis showed that a history of biliary disease was an independent risk factor for ESBL-producing KPLA. Compared with the non-ESBL-producing KPLA group, the ESBL-producing KPLA group had a significantly higher intensive care unit (ICU) admission rate (28.57% vs. 2.33%, p < 0.001). All ESBL-producing KP isolates were susceptible to carbapenems and amikacin. Only the presence of multiloculation on CT was found to be significantly different between the groups (50% vs. 82.56%, p = 0.012). CONCLUSIONS: The presence of biliary disease was an independent risk factor for ESBL-producing KPLA. Patients with ESBL-producing KPLA had a higher ICU admission rate, with only half of patients having evidence of multiloculation on CT.


Assuntos
Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Abscesso Hepático Piogênico/microbiologia , beta-Lactamases/biossíntese , Adulto , Antibacterianos/farmacologia , China/epidemiologia , Feminino , Humanos , Infecções por Klebsiella/diagnóstico por imagem , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/patologia , Klebsiella pneumoniae/efeitos dos fármacos , Abscesso Hepático Piogênico/diagnóstico por imagem , Abscesso Hepático Piogênico/epidemiologia , Abscesso Hepático Piogênico/patologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Tomografia Computadorizada por Raios X
9.
J Infect Public Health ; 13(9): 1330-1335, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32439355

RESUMO

BACKGROUND: Extended-spectrum ß-lactamase producing Enterobacteriaceae (ESBL-PE) and carbapenem-resistant Enterobacteriaceae (CRE) are disseminated worldwide posing a serious public health concern. Although, the presence of ESBL-PE and CRE in Sri Lanka has been reported, the prevalence is unknown. This study aimed to provide up-to-date epidemiological data on multidrug-resistant Enterobacteriaceae and to characterize the molecular determinants of carbapenemase-producing Enterobacteriaceae (CPE) in Sri Lanka. METHODS: A prospective cross-sectional study was conducted at a tertiary care hospital in Sri Lanka between December 2017 and February 2018. ESBL-PE and CRE were identified by disc diffusion method. Carbapenemase production was determined by carbapenem inactivation method and the presence of selected carbapenemase genes were detected by PCR. RESULTS: Five hundred and ninety three Enterobacteriaceae were isolated from variety of clinical samples. Overall prevalence of ESBL-PE and CRE were 26.0% (n = 154) and 9.6% (n = 57), respectively. The highest rate of ESBL-PE (30.8%) was found in urine samples, while the highest occurrence of CRE (20.8%) was seen in respiratory specimens. The most common CRE species identified was K. pneumoniae (n = 46, 80.7%), followed by C. freundii (n = 4, 7.0%), E. coli (n = 3, 5.3%), P. rettgeri (n = 2, 3.5%), E. cloacae (n = 1, 1.7%), and K. aerogenes (n = 1, 1.7%). Carbapenemase production was observed in 54 (94.7%) of CRE isolates. Fifty eight carbapenemase encoding genes were identified in 54 CPE. The most prevalent carbapenemase gene was blaOXA-48-like (n = 48, 88.9%), followed by blaNDM (n = 8, 14.8%), and blaKPC (n = 2, 3.7%). CONCLUSION: This study reports an alarming rate of CRE and the emergence of blaKPC harboring K. pneumoniae in Sri Lanka. The need for preventive measures is highlighted to limit the spread of these difficult-to-treat bacteria in the country.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/enzimologia , Enterobacteriaceae/genética , Klebsiella pneumoniae/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/enzimologia , Enterobacteriáceas Resistentes a Carbapenêmicos/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Carbapenêmicos/farmacologia , Criança , Pré-Escolar , Estudos Transversais , Enterobacteriaceae/efeitos dos fármacos , Feminino , Humanos , Lactente , Recém-Nascido , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/enzimologia , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos Prospectivos , Sri Lanka/epidemiologia , Adulto Jovem , Resistência beta-Lactâmica/genética , beta-Lactamases/genética
10.
BMC Infect Dis ; 20(1): 312, 2020 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-32345218

RESUMO

BACKGROUND: While there is increasing knowledge about the gut microbiome, the factors influencing and the significance of the gut resistome are still not well understood. Infant gut commensals risk transferring multidrug-resistant antibiotic resistance genes (ARGs) to pathogenic bacteria. The rapid spread of multidrug-resistant pathogenic bacteria is a worldwide public health concern. Better understanding of the naïve infant gut resistome may build the evidence base for antimicrobial stewardship in both humans and in the food industry. Given the high carriage rate of extended spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in Asia, we aimed to evaluate community prevalence, dynamics, and longitudinal changes in antibiotic resistance gene (ARG) profiles and prevalence of ESBL-producing E. coli and K. pneumoniae in the intestinal microbiome of infants participating in the Growing Up in Singapore Towards Healthy Outcomes (GUSTO) study, a longitudinal cohort study of pregnant women and their infants. METHODS: We analysed ARGs in the first year of life among 75 infants at risk of eczema who had stool samples collected at multiple timepoints using metagenomics. RESULTS: The mean number of ARGs per infant increased with age. The most common ARGs identified confer resistance to aminoglycoside, beta-lactam, macrolide and tetracycline antibiotics; all infants harboured these antibiotic resistance genes at some point in the first year of life. Few ARGs persisted throughout the first year of life. Beta-lactam resistant Escherichia coli and Klebsiella pneumoniae were detected in 4 (5.3%) and 32 (42.7%) of subjects respectively. CONCLUSION: In this longitudinal cohort study of infants living in a region with high endemic antibacterial resistance, we demonstrate that majority of the infants harboured several antibiotic resistance genes in their gut and showed that the infant gut resistome is diverse and dynamic over the first year of life.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Eczema/diagnóstico , Microbioma Gastrointestinal/efeitos dos fármacos , Aminoglicosídeos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Eczema/etiologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/enzimologia , Escherichia coli/isolamento & purificação , Fezes/microbiologia , Feminino , Humanos , Lactente , Recém-Nascido , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Estudos Longitudinais , Masculino , Risco , beta-Lactamases/genética , beta-Lactamases/metabolismo , beta-Lactamas/farmacologia
11.
Am J Trop Med Hyg ; 102(5): 1137-1143, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32157990

RESUMO

Although there has been an increasing incidence of bacteremia caused by extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-E) across South East Asia, there are sparse data from the Lao PDR, where laboratory capacity for antimicrobial resistance surveillance is limited. We, therefore, retrospectively reviewed bacteremia caused by ESBL-producing Escherichia coli and Klebsiella pneumoniae between 2010 and 2014 at Mahosot Hospital, Vientiane, Lao PDR. Clinical and laboratory data relating to all episodes of ESBL-E bacteremia were reviewed over the 5-year period and compared with non-ESBL-E bacteremia. Blood cultures positive for E. coli or K. pneumoniae were identified retrospectively from laboratory records. Clinical and laboratory data were extracted from research databases and case notes and analyzed using STATA. Between 2010 and 2014, we identified 360 patients with E. coli (n = 249) or K. pneumoniae (n = 111) bacteremia, representing 34.8% of all patients with clinically significant bacteremia. Seventy-two (20%) isolates produced ESBL; E. coli accounted for 15.3% (55/360) and K. pneumoniae for 4.7% (17/360), respectively. The incidence of ESBL-producing E. coli bacteremia rose during the study period. By multiple logistic analysis, reported antibiotic use in the previous week was significantly associated with ESBL positivity (P < 0.001, odds ratio 3.89). Although multiresistant, most ESBL-producing E. coli and K. pneumoniae remained susceptible to meropenem (65/65; 100%) and amikacin (64/65; 98.5%). We demonstrated an alarming increase in the incidence of ESBL-E as a cause of bacteremia in Vientiane during the study period. This has implications for empiric therapy of sepsis in Laos, and ongoing surveillance is essential.


Assuntos
Bacteriemia/microbiologia , Infecções por Escherichia coli/tratamento farmacológico , Escherichia coli/efeitos dos fármacos , Infecções por Klebsiella/tratamento farmacológico , beta-Lactamases/metabolismo , Adolescente , Adulto , Idoso , Antibacterianos/uso terapêutico , Bacteriemia/tratamento farmacológico , Bacteriemia/epidemiologia , Escherichia coli/enzimologia , Infecções por Escherichia coli/epidemiologia , Feminino , Humanos , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/enzimologia , Laos/epidemiologia , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Adulto Jovem , Resistência beta-Lactâmica
12.
Diagn Microbiol Infect Dis ; 97(1): 114996, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32098688

RESUMO

Whole genome sequencing (WGS) is replacing traditional microbiological typing methods for investigation of outbreaks in clinical settings. Here, we used a clinical microbiology laboratory core genome multilocus sequence typing (cgMLST) workflow to analyze 40 isolates of K. pneumoniae which are part of the Antimicrobial Resistance Leadership Group (ARLG) isolate collection, alongside 10 Mayo Clinic K. pneumoniae isolates, comparing results to those of pulsed-field gel electrophoresis (PFGE). Additionally, we used the WGS data to predict phenotypic antimicrobial susceptibility (AST). Thirty-one of 40 ARLG K. pneumoniae isolates belonged to the same PFGE type, all of which, alongside 3 isolates of different PFGE types, formed a large cluster by cgMLST. PFGE and cgMLST were completely concordant for the 10 Mayo Clinic K. pneumoniae isolates. For AST prediction, the overall agreement between phenotypic AST and genotypic prediction was 95.6%.


Assuntos
Antibacterianos/farmacologia , Genoma Bacteriano , Infecções por Klebsiella/diagnóstico , Klebsiella pneumoniae/classificação , Klebsiella pneumoniae/efeitos dos fármacos , Tipagem de Sequências Multilocus , Técnicas de Tipagem Bacteriana , Eletroforese em Gel de Campo Pulsado , Genótipo , Humanos , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/enzimologia , Testes de Sensibilidade Microbiana , Fenótipo , Sequenciamento Completo do Genoma , Fluxo de Trabalho , beta-Lactamases
13.
Int J Infect Dis ; 93: 252-257, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32061861

RESUMO

OBJECTIVES: Evaluate the role of porins in the susceptibility of Klebsiella pneumoniae to ceftaroline and ceftaroline-avibactam. METHODS: Susceptibility to ceftaroline and ceftaroline-avibactam was tested by broth microdilution method in Klebsiella pneumoniae isolates (n = 65), including isogenic mutants (n = 30) and clinical isolates (n = 35), with different outer membrane porin defects in the presence or absence of beta lactamases. RESULTS: Ceftaroline exhibited excellent activity against all the isogenic porin mutants with a MIC range of 0.125-0.25 µg/ml. Ceftaroline showed limited activity in the presence of extended spectrum ß-lactamase enzymes in isogenic mutant constructs as expected but regained effectiveness in combination with avibactam against these isolates except those carrying metallo-carbapenemase (IMP-4) with an MIC range of 0.25->32 µg/ml. Ceftaroline-avibactam was able to inhibit 86% of the clinical isolates (n = 35) of Klebsiella pneumoniae carrying porin defects and multiple beta lactamases with only four isolates showing raised MICs against the combination (MIC range 0.125-4 µg/ml). One clinical isolate with IMP-4 carbapenemase had an MIC value of >32 µg/ml. CONCLUSION: Outer membrane porins play a key role in the transport of ceftaroline inKlebsiella pneumoniae but it remains effective in isolates with altered permeability due to common porin mutations. The addition of avibactam substantially enhances the potency of ceftaroline providing an effective remedy to the problem of omnipresent beta lactamases in these bacteria.


Assuntos
Compostos Azabicíclicos/farmacologia , Proteínas de Bactérias/farmacologia , Cefalosporinas/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , Porinas/genética , beta-Lactamases/farmacologia , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Humanos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/isolamento & purificação , Testes de Sensibilidade Microbiana , Mutação , Porinas/fisiologia , beta-Lactamases/metabolismo
14.
Epidemiol Infect ; 148: e68, 2020 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-32081112

RESUMO

Infections due to extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-E) are often preceded by asymptomatic carriage. Higher incidences in enteric infectious diseases during summer have been reported. Here, we assessed whether the presence of seasonality in intestinal ESBL-Escherichia coli/Klebsiella pneumoniae (ESBL-E/K) carriage in the general Dutch population exists. From 2014 to 2017, the faecal carriage of ESBL-E/K in healthy individuals was determined in three cross-sectional studies in the Netherlands, including 5985 subjects. Results were pooled to identify seasonal trends in prevalence (by month of sampling). Multivariate logistic regression analysis was used to calculate pooled odds ratios and 95% confidence intervals. Results were adjusted for age, sex, antibiotic use and travel. Overall prevalence of ESBL-E/K carriage was 4.3% (n = 260 ESBL-E/K-positive), with differences between months ranging from 2.6% to 7.4%. Compared to January, the monthly prevalence of ESBL-E carriage was highest in August (OR 1.88, 95% CI 1.02-3.49) and September (OR 2.25, 95% CI 1.30-3.89). The observed monthly differences in ESBL-E/K carriage rates suggest that there is seasonal variation in exposure to ESBL-E/K other than due to travelling and antibiotic use. This should be taken into account in designing future ESBL-E prevalence studies in temperate regions.


Assuntos
Portador Sadio , Farmacorresistência Bacteriana , Infecções por Enterobacteriaceae , Escherichia coli , Klebsiella pneumoniae , Adolescente , Adulto , Idoso , Proteínas de Bactérias , Portador Sadio/epidemiologia , Portador Sadio/microbiologia , Estudos Transversais , Infecções por Enterobacteriaceae/epidemiologia , Infecções por Enterobacteriaceae/microbiologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/enzimologia , Escherichia coli/isolamento & purificação , Fezes/microbiologia , Feminino , Humanos , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Prevalência , Estações do Ano , Adulto Jovem , beta-Lactamases
15.
BMC Infect Dis ; 20(1): 166, 2020 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-32087700

RESUMO

BACKGROUND: In this study, we evaluated the genetic relatedness of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-KPN) isolates from an outbreak in a neonatal intensive care unit (NICU) in August 2017, We implemented an active countermeasure to control this outbreak successfully. METHODS: The incidence of healthcare-associated ESBL-KPN bacteremia was evaluated before and after initiating enhanced infection control (IC) practices in January 2018. Surveillance cultures were set up and monitored for neonates, medical personnel, and NICU environments. Molecular analyses, including pulse-field gel electrophoresis (PFGE), sequence typing, and ESBL genotyping, were performed for the isolated KPN strains. RESULTS: After implementing the enhanced IC procedures, the healthcare-associated bacteremia rate decreased from 6.0 to 0.0 per 1000 patient-days. Samples from neonates (n = 11/15, 73.3%), medical personnel (n = 1/41, 2.4%), and medical devices and the environments (6/181, 3.3%) tested positive for ESBL-KPN in the surveillance cultures in December 2017. Active surveillance cultures revealed that 23 of 72 neonates who were screened (31.9%) were colonized with ESBL-KPN between January and March 2018. All the isolates demonstrated closely related PFGE patterns and were identified as ST307 strain carrying the CTX-M-15 gene. CONCLUSIONS: Contaminated NICU environments and medical devices, as well as transmission by medical personnel, appeared to be the source of the outbreak of ESBL-KPN infection. We employed an enhanced IC strategy during January-March 2018 and successfully controlled the clonal outbreak of CTX-M-15-positive KPN. ST307 has emerged as an important bacteremia-causing pathogen in the NICU and should be carefully monitored.


Assuntos
Bacteriemia/epidemiologia , Infecção Hospitalar/epidemiologia , Surtos de Doenças , Controle de Infecções/métodos , Unidades de Terapia Intensiva Neonatal , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/genética , Infecção Hospitalar/mortalidade , Feminino , Genótipo , Humanos , Incidência , Recém-Nascido , Infecções por Klebsiella/mortalidade , Infecções por Klebsiella/transmissão , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Masculino , África do Sul/epidemiologia , beta-Lactamases/biossíntese
16.
Microb Genom ; 6(1)2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-32003322

RESUMO

In this work, we used a whole-genome sequencing (WGS) approach to study the features of KPC-producing Klebsiella pneumoniae (KPC-Kp) spreading in a large Italian long-term acute-care rehabilitation facility (LTACRF), and to track the dynamics of dissemination within this setting. Thirty-eight, non-replicated, KPC-Kp isolates from colonized patients (either already colonized at admission or colonized during admission), collected during 2016, were subjected to antimicrobial-susceptibility testing and WGS. All isolates were resistant to ß-lactams, with the exception of ceftazidime/avibactam (97.4 % susceptible). The second most effective agent was fosfomycin, followed by colistin, trimethoprim/sulfamethoxazole, gentamicin and amikacin (92.1, 86.8, 60.5, 44.7 and 50 % of susceptibility, respectively). A large proportion of isolates (n=18/38, 47.4%) belonged to clonal group (CG) 101, and most of them (n=15) to a new sequence type (ST) designated as ST2502. All the CG101 isolates had a capsule locus type KL17. The ST2502 harboured the genes encoding for the yersiniabactin siderophore and the ArmA methylase, conferring high-level resistance to aminoglycosides. The second most represented lineage of isolates (16/38, 42.1%) belonged to ST512 of CG258. Analysing WGS data, we were able to ascertain the common origin of some isolates imported from other hospitals, and to track several clusters of in-LTACRF cross-transmissions. The results revealed that, in peculiar epidemiological settings such as LTACRF, new KPC-Kp clones different from those prevailing in acute-care hospitals and associated with uncommon resistance and virulence determinants can successfully emerge and disseminate.


Assuntos
Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Farmacorresistência Bacteriana/genética , Doenças Endêmicas , Humanos , Itália/epidemiologia , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/crescimento & desenvolvimento , Assistência de Longa Duração , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Instituições de Cuidados Especializados de Enfermagem , Sequenciamento Completo do Genoma , beta-Lactamases/metabolismo
17.
J Infect Public Health ; 13(4): 647-650, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32067931

RESUMO

Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacteriaceae have been isolated from human patients in many countries across the globe but rarely in Saudi Arabia. Here we provide the genomic characterization of the first KPC-producing K. pneumoniae isolated from the urine of a patient in Riyadh, Saudi Arabia, who had a recent travel history to Egypt involving a medical procedure. Presence of KPC-encoding gene initially detected with the Xpert Carba-R assay was confirmed by traditional PCR. Susceptibility testing using the VITEK 2 system, E-test and microbroth dilution methods showed that the K. pneumoniae isolate, namely SA01_KPC-2, was resistant to all antibiotics except colistin and ceftazidime/avibactam. Whole-genome sequencing (WGS), performed on the Illumina Miseq instrument, identified the isolate as sequence type (ST) 383 and serotype KL30-D1 O1v2. Genome assembly of SA01_KPC-2 indicated the presence of two plasmids. Plasmid pSA01_KPC-2, of approximately 45.9 kb in size, harbored the blaKPC-2 flanked by ISKpn27 and ISKpn6 and had a backbone similar to published KPC-carrying IncX6 plasmids. The second plasmid pSA01_incHIB1, a derivative of published 372-kb plasmid pKpvST383, carried genes encoding virulence factors and resistance to five classes of antibiotics. The isolation of the first KPC producer in Saudi Arabia requires high attention and rapid interventions to prevent further spread.


Assuntos
Proteínas de Bactérias/genética , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/enzimologia , beta-Lactamases/genética , Idoso , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana Múltipla/genética , Humanos , Infecções por Klebsiella/tratamento farmacológico , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/isolamento & purificação , Masculino , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Arábia Saudita/epidemiologia , Sequenciamento Completo do Genoma
18.
J Med Microbiol ; 69(1): 82-86, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31904319

RESUMO

In recent years, Serratia marcescens has emerged as an important agent of hospital-acquired infections, such as pneumonia, urinary tract infection, septicaemia and meningitis, particularly in vulnerable patients. Compared to Klebsiella pneumoniae and Escherichia coli, S. marcescens is less commonly associated with bla KPC genes, yet few cases of plasmid transmission at the gastrointestinal level from K. pneumoniae carbapenemase (KPC)-producing Enterobacterales to S. marcescens have been described. Here we report a case of in vivo acquisition, during a 3-month period of hospitalization in the intensive care unit, of a bla KPC-3 gene carried by a pKpQIL-IT plasmid, and its probable transmission at the bronchial level among different species of Enterobacterales, including K. pneumoniae and S. marcescens. By using whole genome sequence analyses we were able provide insight into the dynamics of carbapenem-resistance determinants acquisition in the lower respiratory tract, a novel anatomical region for such plasmid transmission events, that usually involve the gastrointestinal tract. The co-presence at the same time of both wild-type and resistant Enterobacterales could have been the critical factor leading to the spread of plasmids harbouring carbapenem-resistance genes, of particular importance during surveillance screenings. The possibility of such an event may have significant consequences in terms of antimicrobial treatment, with a potential limitation of therapeutic options, thereby further complicating the clinical management of high-risk critically ill patients.


Assuntos
Proteínas de Bactérias/genética , Transferência Genética Horizontal , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/genética , Plasmídeos , Serratia marcescens/enzimologia , Serratia marcescens/genética , beta-Lactamases/genética , Adulto , Infecção Hospitalar/microbiologia , Humanos , Unidades de Terapia Intensiva , Infecções por Klebsiella/microbiologia , Masculino , Infecções Respiratórias/microbiologia , Infecções por Serratia/microbiologia , Sequenciamento Completo do Genoma
19.
mSphere ; 5(1)2020 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-31915233

RESUMO

Carbapenemases confer resistance to nearly all ß-lactam antibiotics. The extensive spread of carbapenemase-producing multidrug-resistant bacteria contributes significantly to hospital-acquired infections. We have developed a novel protein-based binding assay that identifies KPC ß-lactamases from clinical isolates. We used the protein-protein interaction between KPCs and a soluble ß-lactamase inhibitory protein (BLIP) variant, BLIPK74T/W112D, which specifically inhibits KPCs but not other ß-lactamases. In this assay, BLIPK74T/W112D was allowed to form complexes with KPC-2 in bacterial cell lysates and then extracted using His tag binding resins. We demonstrated the presence of KPC-2 by monitoring the hydrolysis of a colorimetric ß-lactam substrate. Also, to further increase the accuracy of the method, a BLIPK74T/W112D-mediated inhibition assay was developed. The binding and inhibition assays were validated by testing 127 Klebsiella pneumoniae clinical isolates with known genome sequences for the presence of KPC. Our assays identified a total of 32 strains as KPC-2 producers, a result in 100% concordance with genome sequencing predictions. To further simplify the assay and decrease the time to obtain results, the BLIPK74T/W112D protein was tested in combination with the widely used Carba-NP assay. For this purpose, the genome-sequenced K. pneumoniae strains were tested for the presence of carbapenemases with the Carba-NP test with and without the addition of BLIPK74T/W122D The test accurately identified carbapenemase-producing strains and the addition of BLIPK74T/W112D allowed a further determination that the strains contain KPC carbapenemase. Thus, the BLIPK74T/W112D protein is an effective sensor to specifically detect KPC ß-lactamases produced by clinical isolates.IMPORTANCE Infections caused by carbapenem-resistant Enterobacteriaceae are associated with high therapeutic failure and mortality rates. Thus, it is critical to rapidly identify clinical isolates expressing KPC ß-lactamases to facilitate administration of the correct antibiotic treatment and initiate infection control strategies. To address this problem, we developed a protein-based, KPC-specific binding assay in combination with a cell lysate inhibition assay that provided a 100% identification rate of KPC from clinical isolates of known genomic sequence. In addition, this protein sensor was adapted to the Carba-NP assay to provide a rapid strategy to detect KPC-producing isolates that will facilitate informed treatment of critically ill patients.


Assuntos
Bioensaio/métodos , Farmacorresistência Bacteriana Múltipla , Klebsiella pneumoniae/enzimologia , beta-Lactamases/análise , Antibacterianos/farmacologia , Técnicas Biossensoriais , Colorimetria , Infecção Hospitalar/microbiologia , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana , Ligação Proteica , Reprodutibilidade dos Testes , Inibidores de beta-Lactamases/farmacologia , beta-Lactamas/farmacologia
20.
Nat Commun ; 11(1): 388, 2020 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-31959751

RESUMO

Bacterial microcompartments (BMCs) are prokaryotic organelles consisting of a protein shell and an encapsulated enzymatic core. BMCs are involved in several biochemical processes, such as choline, glycerol and ethanolamine degradation and carbon fixation. Since non-native enzymes can also be encapsulated in BMCs, an improved understanding of BMC shell assembly and encapsulation processes could be useful for synthetic biology applications. Here we report the isolation and recombinant expression of BMC structural genes from the Klebsiella pneumoniae GRM2 locus, the investigation of mechanisms behind encapsulation of the core enzymes, and the characterization of shell particles by cryo-EM. We conclude that the enzymatic core is encapsulated in a hierarchical manner and that the CutC choline lyase may play a secondary role as an adaptor protein. We also present a cryo-EM structure of a pT = 4 quasi-symmetric icosahedral shell particle at 3.3 Å resolution, and demonstrate variability among the minor shell forms.


Assuntos
Proteínas de Bactérias/metabolismo , Klebsiella pneumoniae/citologia , Liases/metabolismo , Organelas/ultraestrutura , Proteínas de Bactérias/genética , Colina/metabolismo , Microscopia Crioeletrônica , Loci Gênicos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/ultraestrutura , Liases/genética , Organelas/enzimologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Biologia Sintética
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