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1.
Rev. chil. infectol ; 37(4): 362-370, ago. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1138560

ABSTRACT

Resumen Introducción: Las enterobacterias son una causa principal de infecciones del torrente sanguíneo y su resistencia antimicrobiana se encuentra en aumento. Esto lleva a un incremento de la morbilidad-mortalidad y de los costos en la salud pública. Las enterobacterias resistentes a carbapenems representan un grave desafío a nivel global ya que existen escasas opciones terapéuticas disponibles. Objetivo: Caracterización clínico/microbiológica de las bacteriemias resistentes a carbapenémicos observadas en un período de 4 años. Material y Método: Estudio retrospectivo, observacional y descriptivo, sobre las bacteriemias por enterobacterias resistentes y sensibles a carbapenems. Resultados: Se analizó un total de 84 pacientes con bacteriemia por enterobacterias resistentes y sensibles a carbapenems. Entre las resistentes, observamos una mayor proporción de: tratamiento antimicrobiano previo, hospitalización en unidad de terapia intensiva (UTI), inicio de la bacteriemia en UTI y antecedentes de β-lactamasas de espectro extendido. Además, se detectó un amplio predominio de Klebsiella pneumoniae productor de KPC y una mortalidad atribuible de 52,4%. Discusión: El estudio posibilitó profundizar el conocimiento de una enfermedad emergente de elevada mortalidad, en vistas al diseño y aplicación de estrategias de control de infecciones y de esquemas de tratamiento efectivos adaptados a la epidemiologia local.


Abstract Background: Enterobacteriaceae are a major cause of bloodstream infections and their antimicrobial resistance continues to increase. This leads to higher morbidity-mortality rates and public health costs. Carbapenem-resistant Enterobacteriaceae represent a serious challenge globally, since there are few therapeutic options available. Aim: Clinical/microbiological characterization of the carbapenem-resistant bacteremia observed over a period of 4 years. Methods: Retrospective, observational and descriptive study about bacteremia caused by carbapenem-resistant and susceptible Enterobacteriaceae. Results: A total of 84 patients with bacteremia including carbapenem-resistant and susceptible Enterobacteriaceae were analyzed. We found that patients infected with carbapenem-resistant strains presented a higher proportion of: previous antibiotic treatment, hospitalization in intensive care unit (ICU), onset of the bacteremia during hospitalization in ICU and previous infection with extended-spectrum-beta-lactamase producing Enterobacteriaceae. Additionally, we observed a predominance of KPC-producing Klebsiella pneumoniae and an attributable mortality rate of 52.4%. Discussion: This study allowed for a better understanding of an emerging problem with high mortality, which in turn is useful for the design and adoption of infection control strategies and effective treatment regimens adapted to our local epidemiology.


Subject(s)
Humans , Klebsiella Infections/drug therapy , Klebsiella Infections/epidemiology , Bacteremia/drug therapy , Bacteremia/epidemiology , Argentina/epidemiology , beta-Lactamases , Microbial Sensitivity Tests , Carbapenems/pharmacology , Retrospective Studies , Drug Resistance, Bacterial , Enterobacteriaceae , Klebsiella pneumoniae , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology
2.
Rev. chil. infectol ; 37(4): 389-394, ago. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1138563

ABSTRACT

Resumen Introducción: Pseudomonas aeruginosa es relevante en infecciones asociadas a la atención de salud, principalmente cuando presenta resistencia a carbapenémicos. Objetivos: Estudiar la producción de carbapenemasas en P. aeruginosa, con susceptibilidad disminuida a carbapenémicos procesadas en el Laboratorio de Microbiología de la Red de Salud UC-CHRISTUS entre 2014-2015, y compararlas con las cepas estudiadas en 2004-2005. Métodos: Entre enero de 2014 y junio de 2015, se aislaron 459 cepas de P. aeruginosa provenientes de muestras clínicas. La susceptibilidad fue determinada por dilución en agar y a las cepas con susceptibilidad disminuida a carbapenémicos se les realizó test de carbaNP. Las cepas positivas fueron estudiadas por RPC para genes blaVIM, blaVIM-1, blaVIM-2, blaIMP, blaNDM, blaKPC, blaOXA y blaIMI. Se realizó en cepas seleccionadas electroforesis de campo pulsado. Resultados: De las 459 cepas estudiadas, 300 presentaban susceptibilidad disminuida a carbapenémicos (65,3%). De éstas, 183 fueron viables para estudio, correspondientes a 164 pacientes. El test de carbaNP fue positivo en 44 cepas de las 183 cepas (24%). Los genes de resistencia encontrados fueron: blaVIM-2 en 35 cepas, blaKPC-2+VIM-2 en 7 cepas y blaKPC-2 en 2 cepas. En las cepas blaKPC-2 se encontró relación clonal entre ellas. Conclusiones: Un 65,3% de P. aeruginosa presentó susceptibilidad disminuida a carbapenémicos, observándose que la presencia de carbapenemasas no es el principal mecanismo de resistencia. Además, se describe la emergencia en Chile de cepas de P. aeruginosa con carbapenemasas del tipo KPC-2 sola o en combinación con VIM-2.


Abstract Background: Pseudomonas aeruginosa is a relevant infectious agent affecting patients within health care setting; this situation is worsening with the appearance of strains resistance to carbapenems. Aims: To study carbapenemase production in P. aeruginosa with decreased susceptibility to carbapenems processed in the microbiology laboratory of the Health Network UC-CHRISTUS in 2014-2015 and compare them with the strains studied in 2004-2005. Methods: Between January 2014 and June 2015, 459 strains of P. aeruginosa from clinical samples were isolated. Susceptibility was determined by dilution in agar and strains with reduced susceptibility to carbapenems were tested for carbaNP. Positive strains were studied by PCR for blaVIM, blaVIM-1, blaVIM-2, blaIMP, blaNDM, blaKPC, blaOXA and blaIMI genes. Pulsed field electrophoresis was performed on selected strains. Results: From 459 strains studied, 300 had reduced susceptibility to carbapenems (65.3%). Of these, 183 were viable for study, corresponding to 164 patients. The carbaNP test was positive in 44 strains of the 183 strains (24%). The resistance genes found were: blaVIM-2 in 35 strains, blaKPC-2+VIM-2 in 7 strains and blaKPC-2 in 2 strains. In the blaKPC-2 strains clonal relation between them was found. Conclusions: A 65.3% of P. aeruginosa presented decreased susceptibility to carbapenems being the presence of carbapenemases not the main resistance mechanism. In addition, the emergence in Chile of P. aeruginosa strains with bla of the KPC-2 type alone or in combination with VIM-2 is described.


Subject(s)
Humans , Pseudomonas aeruginosa/genetics , Carbapenems/pharmacology , Bacterial Proteins/genetics , beta-Lactamases , Microbial Sensitivity Tests , Chile , Anti-Bacterial Agents/pharmacology
3.
Rev. Soc. Bras. Med. Trop ; 53: e20200413, 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1136893

ABSTRACT

Abstract Consumption of carbapenem has increased due to extended-spectrum beta-lactamase-producing bacteria spreading. Ertapenem has been suggested as a not carbapenem-resistance inducer. We performed a scoping review of carbapenem-sparing stewardship with ertapenem and its impact on the antibiotic resistance of Gram-negative bacilli. We searched PubMed for studies that used ertapenem as a strategy to reduce resistance to carbapenems and included epidemiologic studies with this strategy to evaluate susceptibility patterns to cephalosporins, quinolones, and carbapenems in Gram-negative-bacilli. The search period included only studies in English, up to February 2018. From 1294 articles, 12 studies were included, mostly from the Americas. Enterobacteriaceae resistance to quinolones and cephalosporins was evaluated in 6 studies and carbapenem resistance in 4 studies. Group 2 carbapenem (imipenem/meropenem/doripenem) resistance on A. baumannii was evaluated in 6 studies. All studies evaluated P. aeruginosa resistance to Group 2 carbapenem. Resistance profiles of Enterobacteriaceae and P. aeruginosa to Group 2 carbapenems were not associated with ertapenem consumption. The resistance rate of A. baumannii to Group 2 carbapenems after ertapenem introduction was not clear due to a lack of studies without bias. In summary, ertapenem as a strategy to spare use of Group 2 carbapenems may be an option to stewardship programs without increasing resistance of Enterobacteriaceae and P. aeruginosa. More studies are needed to evaluate the influence of ertapenem on A. baumannii.


Subject(s)
Carbapenems/pharmacology , Anti-Bacterial Agents/therapeutic use , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests , Drug Resistance, Bacterial , beta-Lactams/pharmacology , Ertapenem
4.
Rev. Soc. Bras. Med. Trop ; 52: e20180502, 2019. tab, graf
Article in English | LILACS | ID: biblio-1041563

ABSTRACT

Abstract INTRODUCTION: Plant products are sources for drug development against multidrug resistant bacteria. METHODS The antimicrobial activity of Origanum vulgare L. essential oil (OVeo) against carbapenem-resistant strains was assessed by disk-diffusion, microdilution (REMA-Resazurin Microtiter Assay), and time kill assays. RESULTS Carbapenemase production was confirmed for all strains. OVeo exhibited a minimum inhibitory concentration of 0.059% v/v for Klebsiella pneumoniae and Serratia marcescens, and of 0.015 % v/v for Acinetobacter baumannii. A decrease in cell count was observed after a 4 h treatment. CONCLUSIONS OVeo antimicrobial effect was rapid and consistent, making it a candidate for developing alternative therapeutic options against carbapenem-resistant strains.


Subject(s)
Humans , Serratia marcescens/drug effects , Oils, Volatile/pharmacology , Acinetobacter baumannii/drug effects , Origanum/chemistry , Gram-Negative Bacteria/drug effects , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/pharmacology , Serratia marcescens/growth & development , Bacterial Proteins , beta-Lactamases , Microbial Sensitivity Tests , Carbapenems/pharmacology , Drug Resistance, Multiple, Bacterial , Acinetobacter baumannii/growth & development , Gram-Negative Bacteria/growth & development , Klebsiella pneumoniae/growth & development , Anti-Bacterial Agents/classification
5.
São Paulo; s.n; s.n; 2019. 69 p. tab, graf.
Thesis in Portuguese | LILACS | ID: biblio-1049564

ABSTRACT

Pseudomonas aeruginosa é um importante agente de infecções relacionadas à assistência à saúde em todo o mundo. O tratamento empírico de infecções graves causadas por esta espécie usualmente inclui os carbapenêmicos. Essa terapia, se inadequada, está relacionada a um aumento significativo da mortalidade. Os principais mecanismos de resistência aos carbapenêmicos em bacilos Gram-negativos são a redução da permeabilidade da membrana externa, hiperexpressão de bombas de efluxo e produção de betalactamases, sendo este último, o mais eficiente. O objetivo deste trabalho consistiu na caracterização de carbapenemases novas ou emergentes e seu contexto genético em P. aeruginosa. Foram utilizados 11 isolados de P. aeruginosa capazes de hidrolisar o imipenem em ensaio espectrofotométrico e negativos para genes de carbapenemases conhecidos e uma cepa produtora de KPC-2. Tais isolados foram submetidos à confirmação do perfil de resistência aos carbapenêmicos pelos métodos de disco-difusão e microdiluição em caldo (CIM), bloqueio enzimático com EDTA e Blue-Carba. O perfil plasmidial foi avaliado utilizando-se os métodos de lise alcalina e eletroforese em campos pulsados (PFGE). Os genomas foram sequenciados utilizando-se o sistema MiSeq. O perfil clonal dos isolados foi avaliado por PFGE e Multilocus Sequence Typing (MLST). Dez isolados apresentaram Blue-Carba negativo, compatível com a presença de carbapenemases fracas. Quatro isolados apresentaram plasmídeos, visualizados em gel de agarose após PFGE. Os plasmídeos do isolado D5303023 foram eletroporados em E. coli TOP 10 e selecionados em meio LB contendo 1µg/mL imipenem, contudo não foram obtidos transformantes. A análise por PFGE identificou sete grupos clonais distintos. Quanto à tipagem por MLST, foi detectado um novo tipo ST3187 e os ST277 e ST1560 foram os predominantes. O isolado D9203039 apresentou uma carbapenemase GES-20 associada a uma betalactamase de espectro estendido GES-19, sendo os genes que as codificam localizados no integron In724. Esse isolado pertence ao ST309, clone de potencial alto risco, associado ao fenótipo de resistência a múltiplos antimicrobianos no México. O genoma da cepa positiva para KPC-2 foi digerido com a S1 nuclease e submetido à PFGE, evidenciando um plasmídeo de aproximadamente 453 kb. Após análise do sequenciamento confirmou-se que a cepa pertence ao ST446 e foi observada a presença do gene blaKPC-2 em um contig de 128.487 bp. Este contig apresentou 99,9% de similaridade com o plasmídeo 1 da cepa P. aeruginosa RW109; no entanto, ensaios de conjugação bacteriana falharam em obter colônias de transconjugantes. O gene blaKPC-2 foi encontrado flanqueado à montante por uma estrutura truncada de um transpóson da família Tn3 e à jusante por uma ISKpn6 truncada. As análises das sequências de DNA das demais cepas evidenciaram a presença de sequências gênicas, que traduzidas, apresentavam similaridade para sete metalobetalactamases (MBL), indicando a potencial presença de novos genes de carbapenemases. Foi caracterizada pela primeira vez no Brasil cepa produtora da carbapenemase GES-20 e o novo ST3187. Foram detectados potenciais novos genes de carbapenemases. O gene blaKPC-2 no isolado J5083553 tem provável localização plasmidial


Pseudomonas aeruginosa is a major agent of healthcare-related infections worldwide. Empirical treatment of serious infections caused by this species usually includes carbapenems. This therapy, if inadequate, is related to a significant increase in mortality. The main mechanisms of carbapenem resistance in Gram-negative bacteria are the reduction of outer membrane permeability, overexpression of efflux pumps and beta-lactamase production, the latter being the most efficient. The aim of this work was the characterization of new or emerging carbapenemases and their genetic context in P. aeruginosa. Eleven P. aeruginosa isolates capable of hydrolyzing imipenem in spectrophotometric assay and negative for known carbapenemases genes were used, as well as a KPC-2 producing strain. These isolates were subjected to confirmation of carbapenem resistance profile by disk diffusion, broth microdilution (MIC) and enzyme inhibition by EDTA and Blue-Carba methods. Plasmid profile was evaluated using alkaline lysis and pulsed field electrophoresis (PFGE) methods. Genomes were sequenced using the MiSeq system. The clonal profile of the isolates was evaluated by PFGE and Multilocus Sequence Typing (MLST). Ten isolates presented negative Blue-Carba, compatible with the presence of weak carbapenemases. Four isolates presented plasmids visualized on agarose gel after PFGE. The plasmids of isolate D5303023 were electroporated into E. coli TOP 10 and selected in LB medium containing 1 µg/mL imipenem, however no transformants were obtained. The PFGE analysis identified 7 distinct clonal groups. As for MLST typing, a new type ST3187 was detected and the ST277 and ST1560 were the predominant types. The genome of the KPC-2 positive strain was digested with S1 nuclease and subjected to PFGE, evidencing a plasmid of approximately 453 kb. Isolate D9203039 presented a GES-20 carbapenemase associated with a GES-19 extended spectrum beta-lactamase. The genes encoding them were located in the In724 integron. This isolate belonged to ST309, a potential high risk clone, associated with the multidrug resistant strain in Mexico. After sequencing it was confirmed that the strain belongs to ST446 and it was observed the presence of the blaKPC-2 gene in a contig of 128,487 bp. This contig was 99.9% similar to plasmid 1 of the P. aeruginosa RW103 strain. However, bacterial conjugation assays failed to obtain transconjugant colonies. The blaKPC-2 gene was found flanked upstream by a truncated structure of a Tn3 family transposon and downstream by a truncated ISKpn6. DNA sequence analysis of the other strains showed the presence of gene sequences, which translated, showed similarity to seven metallo-beta-lactamases (MBL), indicating the potential presence of new carbapenemase genes. It was characterized for the first time in Brazil carbapenemase producing strain GES-20 and the new ST3187. Potential new carbapenemase genes were detected. The blaKPC-2 gene in isolate J5083553 has plasmid localization


Subject(s)
Pseudomonas aeruginosa/metabolism , Carbapenems/pharmacology , Genes/immunology
6.
Med. infant ; 25(4): 299-302, diciembre 2018. tab
Article in Spanish | LILACS | ID: biblio-970392

ABSTRACT

Introducción. La bacteriemia por Pseudomonas aeruginosa (PAE) en niños es infrecuente. Objetivo.Describir las características epidemiológicas, clínicas, microbiológicas y evolutivas en niños con bacteriemia por PAE. Métodos. Estudio de cohorte retrospectivo. Resultados. Se incluyeron 100 pacientes (p). La mediana de edad fue de 27 meses (RIC 6-88).Tenían enfermedad de base: 93 p (93%) y 36 de ellos estaban neutropénicos. Ochenta y cinco p (85%) habían recibido antibióticos en el último mes, 60 (60%) tuvieron procedimientos invasivos previos y 81 (81%) tuvieron internaciones previas. Ingresaron con shock séptico 42 p (42%), 56 p (56%) fueron admitidos en unidad de cuidados intensivos (UCI) y 49 (49%) requirieron ventilación mecánica (VM). La bacteriemia fue primaria en 17 p (17%); asociada a catéter en 15 p (15%) y secundaria en 68 p (68%). El foco más frecuente fue mucocutáneo, 21 p, seguido por el pulmonar, 20 p. El tratamiento empírico fue adecuado en 84 p (84%). La resistencia a uno o más grupos de antibióticos se dio en el 38% de los casos, 11% fueron multirresistentes y 15% fueron resistentes sólo a carbapenemes. Fallecieron 31 p (31%). Pseudomonas aeruginosa resistente a carbapenemes en forma exclusiva o combinada con otros antibióticos se relacionó en esta serie a exposición previa a antibióticos, (p≤0,03), tratamiento empírico inicial inadecuado (p≤0,006) y mayor mortalidad (p≤0,01), prolongación de la internación y del tiempo de tratamiento (p≤0,001)


Introduction. Pseudomonas aeruginosa (PAE) associated bacteremia is uncommon in children. Objective. To describe the epidemiological, clinical, and microbiological features and outcome in children with PAE-associated bacteremia. Methods. A retrospective cohort study. Results. 100 patients (p) were included. Median age was 27 months (IQR 6-88). Overall 93 p (93%) had an underlying disease, 36 of whom had neutropenia. Eighty-five p (85%) had received antibiotics over the previous month, 60 (60%) had undergone previous invasive procedures, and 81 (81%) had been previously admitted. Forty-two p (42%) were admitted because of septic shock, 56 p (56%) were admitted to the intensive care unit (ICU), and 49 (49%) required mechanical ventilation (MV). Seventeen p (17%) had primary bacteremia, 15 p (15%) had catheter-related bacteremia, and 68 p (68%) had secondary bacteremia. The most common focus was mucocutaneous (21 p), followed by pulmonary (20 p). Emperical treatment was adequate in 84 p (84%). Resistance to one or more groups of antibiotics was observed in 38% of the cases; 11% were multiresistant and 15% were only resistant to carbapenems. Thirty-one p (31%) died. In our series, Pseudomonas aeruginosa resistant to carbapenems only or combined with other antibiotics was associated with previous exposition to antibiotics (p≤0.03), inadequate initial emperical treatment (p≤0.006), and higher mortality (p≤0.01), and longer hospital stay and treatment duration (p≤0.001)


Subject(s)
Humans , Infant , Child, Preschool , Child , Adolescent , Pseudomonas aeruginosa/isolation & purification , Pseudomonas aeruginosa/drug effects , Pseudomonas Infections/diagnosis , Pseudomonas Infections/microbiology , Pseudomonas Infections/epidemiology , Bacteremia/microbiology , Bacteremia/mortality , Drug Resistance, Multiple, Bacterial/drug effects , Carbapenems/pharmacology , Prospective Studies , Cohort Studies , Anti-Bacterial Agents/pharmacology
7.
Braz. j. microbiol ; 49(4): 914-918, Oct.-Dec. 2018. tab
Article in English | LILACS | ID: biblio-974286

ABSTRACT

ABSTRACT The global emergence of carbapenemases led to the need of developing new methods for their rapid detection. The aim of this study was to evaluate the performance of the rapid tests for carbapenemase-producing and non-producing Enterobacteriaceae. Carbapenem non-susceptible Enterobacteriaceae from a surveillance study submitted to a multiplex real time PCR for carbapenemase detection were included in this study. The isolates were subjected to the rapid phenotypic tests Carba NP, Blue-Carba and Carbapenem Inactivation Method (CIM). A total of 83 carbapenemase-producing (43) and non-producing (40) isolates were included in the study. The sensitivity/specificity were 62.7%/97.5%, 95.3%/100%, and 74.4%/97.5% for Carba NP, Blue-Carba and CIM, respectively. Both Carba NP and Blue-Carba presented their final results after 75 min of incubation; the final results for CIM were obtained only after 8 h. Failure to detect OXA-370 carbapenemase was the main problem for Carba NP and CIM assays. As the Blue-Carba presented the highest sensitivity, it can be considered the best screening test. Conversely, CIM might be the easiest to perform, as it does not require special reagents. The early detection of carbapenemases aids to establish infection control measures and prevent carbapenemases to spread reducing the risk of healthcare associated infections and therapeutic failure.


Subject(s)
Humans , Bacterial Proteins/analysis , beta-Lactamases/analysis , Enterobacteriaceae/enzymology , Enterobacteriaceae Infections/microbiology , Enzyme Assays/methods , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , beta-Lactamases/genetics , beta-Lactamases/metabolism , Brazil , Carbapenems/pharmacology , Polymerase Chain Reaction , Sensitivity and Specificity , Enterobacteriaceae/isolation & purification , Enterobacteriaceae/drug effects , Enterobacteriaceae/genetics , Enterobacteriaceae Infections/diagnosis , Anti-Bacterial Agents/pharmacology
8.
Braz. j. microbiol ; 49(4): 885-890, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974312

ABSTRACT

ABSTRACT In this study, the performance of the "RESIST-3 O.K.N. K-SeT" (Coris BioConcept, Gembloux, Belgium) immunochromatographic assay was evaluated in 132 Klebsiella pneumoniae comprising 102 carbapenem resistant and 30 carbapenem susceptible isolates. Genotypically known isolates of Gram negative bacteria (n = 22) including various species were also tested by the assay as controls. The isolates tested by the immunochromatographic assay and also were run PCR for bla KPC, bla IMP, bla VIM, bla NDM, and bla OXA-48. The rates of bla NDM, bla OXA-48, and bla KPC in carbapenem resistant isolates were found at 52.9%, 39.2%, and 2.0%, respectively. Both bla NDM and bla OXA-48 were found in six (5.9%) isolates. The results of the assay showed 100% concordance with those obtained by PCR in 132 K. pneumoniae. The agreement between the two methods was found to be identical at the isolate level. The assay also correctly detected all genotypically known isolates of Escherichia coli, Serratia marcescens, Citrobacter freundii, Enterobacter cloacae, K. pneumoniae carrying bla KPC, bla NDM, and/or bla OXA-48. On the other hand, the assay did not exhibit any cross-reaction in control isolates harboring bla IMP and bla VIM. We conclude that the RESIST-3 O.K.N. K-SeT is a reliable, rapid, and user friendly test and we recommend it for routine diagnostic laboratories.


Subject(s)
Humans , Bacterial Proteins/analysis , beta-Lactamases/analysis , Klebsiella Infections/microbiology , Immunoassay/methods , Klebsiella pneumoniae/enzymology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Turkey , beta-Lactamases/metabolism , Carbapenems/pharmacology , Polymerase Chain Reaction , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/chemistry , Anti-Bacterial Agents/pharmacology
9.
Braz. j. microbiol ; 49(1): 5-6, Jan.-Mar. 2018.
Article in English | LILACS | ID: biblio-889197

ABSTRACT

ABSTRACT The type strain SUR2 of the novel species Chryseobacterium limigenitum was isolated from a dehydrated sludge of the municipal sewage treatment plant in Dogoše near Maribor in Slovenia. The draft genome, with 60 contigs, 4,697,725 bp, 34.4% of G+C content, was obtained using the Illumina HiSeq 2500-1 platform. Joint Genome Institute Microbial Genome Annotation Pipeline (MGAP v.4) has identified 4322 protein-coding sequences including resistance genes against arsenic and other heavy metals. In addition, a subclass B3 metallo-β-lactamase, which confers resistance to penicillins, cephalosporins and carbapenems, was also present in the genome. The genome sequence provides important information regarding bioremediation potential and pathogenic properties of this newly identified species.


Subject(s)
Sewage/microbiology , Genome, Bacterial , Chryseobacterium/genetics , Penicillins/pharmacology , Phylogeny , Sewage/chemistry , Base Composition , DNA, Bacterial/genetics , Molecular Sequence Data , Base Sequence , Microbial Sensitivity Tests , Carbapenems/pharmacology , Chryseobacterium/isolation & purification , Chryseobacterium/classification , Chryseobacterium/drug effects , Anti-Bacterial Agents/pharmacology
10.
Braz. j. infect. dis ; 22(1): 47-50, Jan.-feb. 2018. tab, graf
Article in English | LILACS | ID: biblio-1039209

ABSTRACT

ABSTRACT Carbapenemases have great importance in the global epidemiological scenario since infections with carbapenemase-producing bacteria are associated with high mortality, especially in hospitalized patients in intensive care units. This study describes two microorganisms producers of the New Delhi Metallo-b-lactamase, Klebsiella pneumoniae and Citrobacter freundii, from two patients admitted to a public hospital in Salvador, Bahia. These are the first clinical cases of New Delhi Metallo-b-lactamase described in microorganisms in the north and northeast Brazil. The isolates were characterized by antimicrobial susceptibility test, with resistance to all β-lactams including carbapenems, negative Modified Hodge Test and the synergy test with Ethylenediaminetetraacetic acid, Phenylboronic Acid and Cloxacillin was positive only with Ethylenediaminetetraacetic acid (difference of >5 mm in the inhibition zone between the disk without and with the inhibitor). Analysis by multiplex PCR for blaIMP, blaVIM, blaNDM, blaKPC and blaOXA-48 enzymes confirmed the presence of blaNDM gene. This report of two different New Delhi Metallo-b-lactamase-producing microorganisms in a different region of Brazil confirms the risk of spreading resistance genes between different species and emphasizes the need for prevention and control of infections caused by these pathogens, which have limited treatment options and have been linked to high mortality rates.


Subject(s)
Humans , Male , Adult , Aged , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Enterobacteriaceae/enzymology , Enterobacteriaceae Infections/microbiology , Bacterial Proteins/drug effects , beta-Lactamases/drug effects , Brazil , Carbapenems/pharmacology , Fatal Outcome , Drug Resistance, Bacterial , Enterobacteriaceae/isolation & purification , Enterobacteriaceae/drug effects , Multiplex Polymerase Chain Reaction , Hospitals, Public
11.
Braz. j. infect. dis ; 22(1): 51-54, Jan.-feb. 2018. tab, graf
Article in English | SES-SP, LILACS, SES-SP, SESSP-IDPCPROD, SES-SP | ID: biblio-1039210

ABSTRACT

ABSTRACT A retrospective cohort study, were evaluated: polymyxin B plus aminoglycosides or polymyxin B plus other antibiotics. Any degree of acute kidney injury occurred in 26 (86.6%) patients. The median time to acute kidney injury was 6.0 (95% CI 3-14) days in the polymyxin-aminoglycoside containing regimen group, against 27.0 (95% CI 6-42) days in the polymyxin with other antimicrobial combinations group (p = 0.03). Polymyxin B with aminoglycosides group progressed faster to any degree of renal dysfunction.


Subject(s)
Humans , Male , Female , Polymyxin B/therapeutic use , Enterobacteriaceae Infections/drug therapy , Kidney/drug effects , Mediastinitis/microbiology , Mediastinitis/drug therapy , Anti-Bacterial Agents/therapeutic use , Microbial Sensitivity Tests , Carbapenems/pharmacology , Retrospective Studies , Treatment Outcome , Statistics, Nonparametric , Risk Assessment , beta-Lactam Resistance/drug effects , Enterobacteriaceae Infections/mortality , Kaplan-Meier Estimate , Acute Kidney Injury/chemically induced , Aminoglycosides/therapeutic use , Mediastinitis/mortality
12.
Clinics ; 72(10): 642-644, Oct. 2017.
Article in English | LILACS | ID: biblio-1039534

ABSTRACT

OBJECTIVE: We describe an IncX4 pHC891/16mcr plasmid carrying mcr-1 in a colistin-resistant and carbapenem-susceptible E. coli isolate (HC891/16), ST156, which caused a blood infection in a Brazilian patient with gallbladder adenocarcinoma. METHODS: Strain HC891/16 was subjected to whole genome sequencing using the MiSeq Platform (Illumina, Inc., USA). Assembly was performed using Mira and ABACAS. RESULTS: The isolates showed resistance only to ciprofloxacin, ampicillin and cefoxitin, and whole-genome sequencing revealed the presence of aac(6')Ib-cr and blaTEM1. CONCLUSION: Our findings warn of the possible silent dissemination of colistin resistance by carbapenem-susceptible mcr-1 producers, as colistin susceptibility is commonly tested only among carbapenem-resistant isolates.


Subject(s)
Humans , Female , Aged , Carbapenems/pharmacology , Bacteremia/drug therapy , Colistin/pharmacology , Escherichia coli Proteins/drug effects , Escherichia coli/drug effects , Anti-Bacterial Agents/pharmacology , Plasmids/drug effects , Brazil , Microbial Sensitivity Tests , Escherichia coli Proteins/isolation & purification , Escherichia coli Proteins/genetics , Drug Resistance, Multiple, Bacterial , Escherichia coli/isolation & purification , Escherichia coli/genetics , Escherichia coli Infections/drug therapy
13.
Braz. j. microbiol ; 48(3): 493-498, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-889153

ABSTRACT

Abstract Carbapenems are considered last-line agents for the treatment of serious infections caused by Klebsiella pneumoniae, and this microorganism may exhibit resistance to β-lactam antibiotics due to different mechanisms of resistance. We evaluated 27 isolates of K. pneumoniae resistant to carbapenems recovered from inpatients at the University Hospital of Santa Maria-RS from July 2013 to August 2014. We carried out antimicrobial susceptibility, carbapenemase detection, testing for the presence of efflux pump by broth microdilution and loss of porin by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Genetic similarity was evaluated by ERIC-PCR. High levels of resistance were verified by the minimum inhibitory concentration for the antimicrobials tested. The blaKPC gene was present in 89% of the clinical isolates. Blue-Carba and combined disk with AFB tests showed 100% concordance, while the combined disk test with EDTA showed a high number of false-positives (48%) compared with the gold-standard genotypic test. Four isolates showed a phenotypic resistance profile consistent with the overexpression of the efflux pump, and all clinical isolates had lost one or both porins. The ERIC-PCR dendrogram demonstrated the presence of nine clusters. The main mechanism of resistance to carbapenems found in the assessed isolates was the presence of the blaKPC gene.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Carbapenems/pharmacology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Drug Resistance, Bacterial , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/metabolism , Microbial Sensitivity Tests
14.
Rev. Soc. Bras. Med. Trop ; 50(2): 173-178, Mar.-Apr. 2017. tab
Article in English | LILACS | ID: biblio-842833

ABSTRACT

Abstract INTRODUCTION: In this study, we used phenotypic methods to screen carbapenem-resistant Enterobacteriaceae (CREs) and evaluated their antimicrobial sensitivity profile. METHODS: One hundred and seventy-eight CREs were isolated at a university hospital in south Brazil in a one-year period. Samples were assessed using disk diffusion tests with inhibitors of β-lactamases such as phenylboronic acid (AFB), cloxacillin (CLOXA), and ethylenediaminetetraacetic acid (EDTA). Strains with differences in zone diameters ≥ 5mm for disks supplemented or not were considered producers of carbapenemases. RESULTS: Klebsiella pneumoniae was the most prevalent CRE, which appeared in 80.3% cases (n = 143). Among clinical materials, the rectal swab was responsible for 43.4% of the isolations (n = 62), followed by urine (18.9%; n = 27). Among the CREs identified in this study, the growth of 56.7% (n = 101) isolates, which were putative producers of Klebsiella pneumoniae carbapenemase (KPC), were inhibited by AFB, whereas 7.3% (n = 13) isolates were inhibited by both AFB and CLOXA and were considered as putative producers of plasmid-mediated AmpC; approximately 3.4% (n = 6) were inhibited by EDTA, which possibly produced metallo-β-lactamase. Lastly, 32.6% (n = 58) cases showed negative results for AFB, CLOXA, and EDTA sensitivity, and represented another class of β-lactamases and/or mechanism of resistance. CONCLUSIONS: Phenotypic screening of CREs is important for clinical laboratories that monitor outbreaks of resistant microbes. Phenotypic tests that use carbapenemase inhibitors and enhancers such as AFB, CLOXA, and EDTA are necessary since they are good screening methods for the detection of carbapenemases.


Subject(s)
Humans , Carbapenems/pharmacology , beta-Lactam Resistance/genetics , Enterobacteriaceae/drug effects , Anti-Bacterial Agents/pharmacology , Phenotype , Microbial Sensitivity Tests , Enterobacteriaceae/classification , Enterobacteriaceae/enzymology , Enterobacteriaceae/genetics , Hospitals, University
15.
Rev. Soc. Bras. Med. Trop ; 50(2): 167-172, Mar.-Apr. 2017. tab
Article in English | LILACS | ID: biblio-842841

ABSTRACT

Abstract INTRODUCTION: Acinetobacter baumannii is a major pathogen causing infections in intensive care units (ICUs). In this study, we aimed to evaluate the presence of A. baumannii in an ICU environment and gloves from ICU workers and to characterize the antimicrobial resistance of the isolates in comparison with those isolated from ICU patients at the same hospital. METHODS: ICU samples were collected from March to November 2010. Isolates biochemically characterized as Acinetobacter calcoaceticus-Acinetobacter baumannii complex were evaluated by PCR targeting the 16S rDNA and bla OXA-51 genes. Antimicrobial susceptibility was determined using the disk diffusion method, and carbapenem-resistant isolates were also evaluated for the minimum inhibitory concentration of imipenem using broth microdilution. The presence of the bla OXA-23 gene was evaluated in isolates with reduced susceptibility to carbapenems. RESULTS: A. baumannii was detected in 9.5% (84) of the 886 samples collected from the ICU environment, including from furniture, medical devices, and gloves, with bed rails being the most contaminated location (23.8%; 20/84). Multidrug-resistant (MDR) A. baumannii was found in 98.8% (83/84) of non-clinical and 97.8% (45/46) of clinical isolates. Reduced susceptibility to carbapenems was detected in 83.3% (70/84) of non-clinical and 80.4% (37/46) of clinical isolates. All isolates resistant to carbapenems harbored bla OXA-23. CONCLUSIONS: We found a strong similarity between the antimicrobial susceptibility profiles of non-clinical and clinical A. baumannii isolates. Such data highlight the ICU environment as a potential origin for the persistence of MDR A. baumannii, and hence the ICU may be a source of hospital-acquired infections caused by this microorganism.


Subject(s)
Humans , Carbapenems/pharmacology , Polymerase Chain Reaction , Gloves, Protective/microbiology , Acinetobacter baumannii/drug effects , Environmental Microbiology , Equipment and Supplies, Hospital/microbiology , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Acinetobacter baumannii/isolation & purification , Acinetobacter baumannii/enzymology , Disk Diffusion Antimicrobial Tests
17.
Braz. j. infect. dis ; 21(1): 57-62, Jan.-Feb. 2017. tab, graf
Article in English | LILACS | ID: biblio-839184

ABSTRACT

Abstract The mechanisms involved in the uncommon resistance phenotype, carbapenem resistance and broad-spectrum cephalosporin susceptibility, were investigated in 25 Pseudomonas aeruginosa clinical isolates that exhibited this phenotype, which were recovered from three different hospitals located in São Paulo, Brazil. The antimicrobial susceptibility profile was determined by CLSI broth microdilution. β-lactamase-encoding genes were investigated by PCR followed by DNA sequencing. Carbapenem hydrolysis activity was investigated by spectrophotometer and MALDI-TOF assays. The mRNA transcription level of oprD was assessed by qRT-PCR and the outer membrane proteins profile was evaluated by SDS-PAGE. Genetic relationship among P. aeruginosa isolates was assessed by PFGE. Carbapenems hydrolysis was not detected by carbapenemase assay in the carbapenem-resistant and cephalosporin-susceptible P. aueruginosa clinical isolates. OprD decreased expression was observed in all P. aeruginosa isolates by qRT-PCR. The outer membrane protein profile by SDS-PAGE suggested a change in the expression of the 46 kDa porin that could correspond to OprD porin. The isolates were clustered into 17 genotypes without predominance of a specific PFGE pattern. These results emphasize the involvement of multiple chromosomal mechanisms in carbapenem-resistance among clinical isolates of P. aeruginosa, alert for adaptation of P. aeruginosa clinical isolates under antimicrobial selective pressure and make aware of the emergence of an uncommon phenotype among P. aeruginosa clinical isolates.


Subject(s)
Humans , Pseudomonas aeruginosa/drug effects , Carbapenems/pharmacology , Cephalosporins/pharmacology , beta-Lactam Resistance/genetics , Anti-Bacterial Agents/pharmacology , Phenotype , Pseudomonas aeruginosa/enzymology , Pseudomonas aeruginosa/genetics , Spectrophotometry, Ultraviolet , Bacterial Outer Membrane Proteins , Bacterial Proteins/metabolism , beta-Lactamases/metabolism , Brazil , DNA, Bacterial , Microbial Sensitivity Tests , Electrophoresis, Gel, Pulsed-Field , Sequence Analysis, DNA , Porins/metabolism , Reverse Transcriptase Polymerase Chain Reaction
18.
Braz. j. microbiol ; 47(4): 785-792, Oct.-Dec. 2016. tab
Article in English | LILACS | ID: biblio-828193

ABSTRACT

Abstract Acinetobacter baumannii is widely recognized as an important pathogen associated with nosocomial infections. The treatment of these infections is often difficult due to the acquisition of resistance genes. A. baumannii presents a high genetic plasticity which allows the accumulation of these resistance determinants leading to multidrug resistance. It is highlighted the importance of the horizontal transfer of resistance genes, through mobile genetic elements and its relationship with increased incidence of multidrug resistant A. baumannii in hospitals. Considering that resistance to carbapenems is very important from the clinical and epidemiological point of view, the aim of this article is to present an overview of the current knowledge about genetic elements related to carbapenem resistance in A. baumannii such as integrons, transposons, resistance islands and insertion sequences.


Subject(s)
DNA, Bacterial , DNA Transposable Elements , Carbapenems/pharmacology , beta-Lactam Resistance , Acinetobacter baumannii/drug effects , Acinetobacter baumannii/genetics , Anti-Bacterial Agents/pharmacology , Mutagenesis, Insertional , Integrons , Genomic Islands
19.
Rev. chil. infectol ; 33(5): 519-523, oct. 2016. ilus, graf, tab
Article in Spanish | LILACS | ID: biblio-844403

ABSTRACT

In order to study the clonal relationship and blaKPC gene detection in clinical isolates of Klebsiella pneumoniae resistant to carbapenems, we analyzed 22 clinical isolates of K. pneumoniae with resistance to imipenem and/ or meropenem, isolated in the laboratory of bacteriology at the University Hospital "Antonio Patricio de Alcalá" (HUAPA) from the Cumana city, Sucre state, Venezuela, for a period of five consecutive years. Susceptibility to different antimicrobials was determined, and the presence of carbapenemases was detected by modified Hodge method, phenyl boronic acid synergy and combination discs. blaKPC gene detection was conducted by polymerase chain reaction and the clonal relationship was determined by pulsed field electrophoresis. High rates of antimicrobial resistance were found, five strains were negative, at least one phenotypic method, and all carried the blaKPC gene. Clonal spread was observed only in the intensive care unit (ICU), while in other services, polyclonality was found. We concluded that blaKPC gene is present in K. pneumoniae strains resistant to carbapenems isolated in the HUAPA and clonal spread it was only in the ICU.


Con el objetivo de estudiar la relación clonal y detección del gen blaKPC en aislados clínicos de Klebsiella pneumoniae resistentes a carbapenémicos, se analizaron 22 cepas clínicas de K. pneumoniae con resistencia a imipenem y/o meropenem, aisladas en el laboratorio de bacteriología del Hospital Universitario "Antonio patricio de Alcalá" (HUAPA) de la ciudad de cumaná, Estado Sucre, Venezuela, durante un período de cinco años continuos. Se determinó la susceptibilidad a diversos antimicrobianos, y se detectó la presencia de carbapenemasas por los métodos de Hodge modificado, sinergia con ácido fenil borónico y combinación de discos. La detección del gen blaKPC se llevó a cabo mediante la técnica de reacción de polimerasa en cadena y la determinación de la relación clonal se realizó por electroforesis de campo pulsado. Se encontraron elevados porcentajes de resistencia antimicrobiana, cinco cepas resultaron negativas, al menos, a un método fenotípico y todas portaban el gen blaKPC. Se observó diseminación de clones únicamente en la Unidad de cuidados Intensivos (UCI), mientras que, en otros servicios, se halló policlonalidad. Se concluye que el gen blaKPC se encuentra presente en cepas de K. pneumoniae resistentes a carbapenémicos aisladas en el HUAPA y que hubo diseminación clonal sólo en UCI.


Subject(s)
Humans , beta-Lactamases/genetics , Klebsiella Infections/microbiology , Carbapenems/pharmacology , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Venezuela , DNA, Bacterial/genetics , Microbial Sensitivity Tests , Polymerase Chain Reaction , Electrophoresis, Gel, Pulsed-Field , Clone Cells
20.
Mem. Inst. Oswaldo Cruz ; 111(9): 551-558, Sept. 2016. tab, graf
Article in English | LILACS | ID: lil-794722

ABSTRACT

Carbapenem-resistance mechanisms are a challenge in the treatment of Pseudomonas aeruginosa infections. We investigated changes in P. aeruginosa carbapenem-resistance determinants over a time period of eight years after the emergence of São Paulo metallo-β-lactamase in a university hospital in Rio de Janeiro, Brazil. Patients admitted to the intensive care unit (ICU) were screened for P. aeruginosa colonisation and followed for the occurrence of infections from April 2007 to April 2008. The ICU environment was also sampled. Isolates were typed using random amplified polymorphic DNA, pulsed-field gel electrophoresis and multilocus sequence typing. Antimicrobial susceptibility was determined by disk diffusion and E-test, production of carbapenemases by a modified-CarbaNP test and presence of carbapenemase-encoding genes by polymerase chain reaction. Non-carbapenemase resistance mechanisms studied included efflux and AmpC overexpression by PAβN and cloxacillin susceptibility enhancement, respectively, as well as oprD mutations. From 472 P. aeruginosa clinical isolates (93 patients) and 17 isolates from the ICU environment, high genotypic diversity and several international clones were observed; one environment isolate belonged to the blaSPM-1 P. aeruginosa epidemic genotype. Among isolates from infections, 10 (29%) were carbapenem resistant: none produced carbapenemases, three exhibited all non-carbapenemase mechanisms studied, six presented a combination of two mechanisms, and one exclusively displayed oprD mutations. Carbapenem-resistant P. aeruginosa displayed a polyclonal profile after the SPM-1 epidemic genotype declined. This phenomenon is connected with blaSPM-1 P. aeruginosa replaced by other carbapenem-resistant pathogens.


Subject(s)
Humans , beta-Lactam Resistance/genetics , beta-Lactamases/biosynthesis , Carbapenems/pharmacology , Pseudomonas aeruginosa/enzymology , Pseudomonas Infections/microbiology , Anti-Bacterial Agents/pharmacology , beta-Lactam Resistance/drug effects , Disk Diffusion Antimicrobial Tests , DNA, Bacterial/genetics , Electrophoresis, Gel, Pulsed-Field , Genotype , Hospitals, University , Intensive Care Units , Multilocus Sequence Typing , Polymerase Chain Reaction , Prospective Studies , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/genetics
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