Detection of SPM and IMP metallo-beta-lactamases in clinical specimens of Pseudomonas aeruginosa from a Brazilian public tertiary hospital

Phenotypic and genotypic SPM and IMP metallo-beta-lactamases (MBL) detection and also the determination of minimal inhibitory concentrations (MIC) to imipenem, meropenem and ceftazidime were evaluated in 47 multidrug-resistant Pseudomonas aeruginosa isolates from clinical specimens. Polymerase chain reaction detected 14 positive samples to either bla(SPM) or bla(IMP) genes, while the best phenotypic assay (ceftazidime substrate and mercaptopropionic acid inhibitor) detected 13 of these samples. Imipenem, meropenem and ceftazidime MICs were higher for MBL positive compared to MBL negative isolates. We describe here the SPM and IMP MBL findings in clinical specimens of P. aeruginosa from the University Hospital of Botucatu Medical School, São Paulo, Brazil, that reinforce local studies showing the high spreading of bla(SPM) and bla(IMP) genes among Brazilian clinical isolates.

Benomyl sensitivity of isolates of Colletotrichum acutatum and C. gloeosporioides from citrus

Postbloom fruit drop (PFD) of citrus, caused by Colletotrichum acutatum, produces orange-brown lesions on petals and results in premature fruit drop and the retention of calyces. C. gloeosporioides is common in groves and causes postharvest anthracnose on fruit. Both diseases are controlled effectively by the fungicide benomyl in research fields and commercial orchards. Highly sensitive and resistant isolates of C. gloeosporioides were found, whereas all isolates of C. acutatum tested were moderately resistant. In preliminary studies conducted in vitro with three isolates of each, mycelial growth of sensitive isolates of C. gloeosporioides was inhibited completely by benomyl (Benlate 50 WP) at 1.0 μg/ml, whereas resistant isolates grew well at 10 μg/ml. Growth of all isolates of C. acutatum was inhibited by about 55% at 0.1 μg/ml and by 80% at 1.0 μg/ml. Spore germination of C. acutatum was inhibited more at 0.1 μg/ml than at 1.0 μg/ml or higher concentrations. In all, 20 isolates of C. acutatum from 17 groves and 20 isolates of C. gloeosporioides from 7 groves were collected from locations with different histories of benomyl usage in São Paulo, Brazil, and Florida, United States. Benomyl at 1.0 μ.g/ml completely inhibited growth of 133 isolates of C. gloeosporioides, with the exception of 7 isolates that were highly resistant to the fungicide, whereas all isolates of C. acutatum were only partially inhibited at 0.1 and 1.0 μg/ml. Analysis of variance indicated that the sensitivity of the isolates of C. acutatum was not affected by benomyl usage or grove of origin, and country of origin had only minor effects. No highly resistant or sensitive isolate of C. acutatum was recovered. Partial sequencing of the β-tubulin gene did not reveal nucleotide substitutions in codons 198 or 200 in C. acutatum that usually are associated with benomyl resistance in other fungi.

Detection of SPM and IMP metallo-β-lactamases in clinical specimens of Pseudomonas aeruginosa from a Brazilian public tertiary hospital

Phenotypic and genotypic SPM and IMP metallo-β-lactamases (MBL) detection and also the determination of minimal inhibitory concentrations (MIC) to imipenem, meropenem and ceftazidime were evaluated in 47 multidrug-resistant Pseudomonas aeruginosa isolates from clinical specimens. Polymerase chain reaction detected 14 positive samples to either blaSPM or blaIMP genes, while the best phenotypic assay (ceftazidime substrate and mercaptopropionic acid inhibitor) detected 13 of these samples. Imipenem, meropenem and ceftazidime MICs were higher for MBL positive compared to MBL negative isolates. We describe here the SPM and IMP MBL findings in clinical specimens of P. aeruginosa from the University Hospital of Botucatu Medical School, São Paulo, Brazil, that reinforce local studies showing the high spreading of blaSPM and blaIMP genes among Brazilian clinical isolates. © 2011 Elsevier Editora Ltda.

Colonization pressure and risk factors for acquisition of imipenem-resistant Acinetobacter baumannii in a medical surgical intensive care unit in Brazil

We studied a nonconcurrent cohort of 582 patients admitted to a medical-surgical intensive care unit. Use of antimicrobials (imipenem and metronidazole) was a risk factor for acquisition of imipenem-resistant Acinetobacter baumannii only for the subcohort of patients admitted in months in which colonization pressure was lower than the median value. © 2013 by the Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Detecção e identificação de genes de β-lactamases de espectro estendido em Salmonella spp. e Escherichia coli isoladas de carnes de frango, suína e bovina destinadas ao consumo humano

Pós-graduação em Microbiologia - IBILCE

Análise de cepas de Escherichia coli Shiga toxigênica (STEC) e E. coli patogênica extraintestinal (ExPEC) isoladas de cachorros diarréicos atendidos em clínica privada no município de Ituverava, SP

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Construção de um painel com isolados clínicos de Mycobacterium tuberculosis com genes de resistência a quimioterápicos, para o estudo de novas drogas anti-TB

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Atividade antibacteriana de plantas medicinais frente á bactérias multirresistentes e a sua interação com drogas antimicrobianas

O controle de micro-organismos infecciosos multirresistentes às vezes é ineficaz mesmo com o desenvolvimento de novos antibióticos. Diversos extratos de plantas medicinais têm efeitos antimicrobianos o que pode representar uma alternativa terapêutica para doenças infecciosas, principalmente quando associados aos antibióticos de uso clínico. O objetivo do trabalho foi avaliar a atividade antibacteriana de plantas medicinais sobre bactérias multirresistentes e os efeitos de sua interação com drogas antimicrobianas. Foi determinada a atividade antibacteriana de extratos e frações das plantas Eleutherine plicata (marupazinho), Geissospermum vellosii (pau-pereira) e Portulaca pilosa (amor-crescido) frente a isolados de Staphylococcus aureus Oxacilina Resistente (ORSA) e de Pseudomonas aeruginosa multirresistente, provenientes de processos clínicos humanos, assim como a interação destes produtos vegetais com drogas antimicrobianas de uso clínico. A atividade antibacteriana foi determinada pelo método de disco difusão em ágar Muller Hinton e a Concentração Inibitória Mínima (CIM) pela técnica de microdiluição em placas utilizando caldo Muller Hinton como meio de cultura e resazurina a 0,01% como revelador de crescimento bacteriano. Os extratos e frações foram testados nas concentrações de 500, 250, 125, 62,5, 31,2 e 16,2 μg/mL dissolvidos em DMSO a 10%. As plantas E. plicata e G. vellosii demonstraram atividade contra os isolados ORSA com CIM de 125 μg/mL, enquanto que P. pilosa teve ação sobre os isolados de P. aeruginosa multirresistentes com CIM de 250 μg/mL. Ocorreram 25% de sinergismo e apenas 5% de antagonismo entre as 120 interações de produtos vegetais e drogas antimicrobianas testadas. Frente aos isolados ORSA houve sinergismo com as drogas ciprofloxacina, clindamicina e vancomicina tanto com os derivados de E. plicata como os de G. vellosii. Os produtos de P. pilosa potencializaram a ação das drogas aztreonam, cefepime e piperacilina+tazobactam frente aos isolados de P. aeruginosa multirresistentes. Os resultados comprovaram o potencial das plantas E. plicata, G. vellosii e P. pilosa no controle de infecções bacterianas envolvendo fenótipos multidrogas resistentes (MDR) e que a sua interação com drogas antibacterianas pode representar uma nova alternativa na terapia destas infecções.

Staphylococcus epidermis e Staphylococcus haemolycus: detecção de genes de biofilme, toxinas, resistência a antimicrobianos e tipagem clonal em isolados de hemoculturas

Pós-graduação em Biologia Geral e Aplicada - IBB

First Baseline of Circulating Genotypic Lineages of Mycobacterium tuberculosis in Patients from the Brazilian Borders with Argentina and Paraguay

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Risk factors for the acquisition of imipenem-resistant Acinetobacter baumannii in a burn unit: An appraisal of the effect of colonization pressure

Imipenem-resistant Acinetobacter baumannii (IRAB) is a major threat for critically ill patients, including those admitted to burn units. Recent studies have suggested that colonization pressure (the proportion of patients or patient-days harbouring the pathogen of interest) is an important driver of the risk for acquisition of multidrug-resistant organisms. With that in mind, we conducted a cohort study, enrolling 208 patients admitted to a burn unit from November 2008 through December 2009. The outcome of interest was the acquisition of IRAB. In addition to the usual risk factors, we assessed the impact of colonization pressure. The number of wound excisions (odds ratio (OR) 12.06, 95% confidence interval (CI) 2.82-51.64) and the number of antimicrobials used (OR 22.82, 95% CI 5.15-101.19) were significant risk factors for the outcome of interest. On the other hand, colonization pressure (measured for whole time of exposure or up to the last 14, 7, or 3 days) was not associated with the risk for IRAB acquisition.

Investigação de genes de resistência a carbapenêmicos e aminoglicosídeos e tipagem molecular de amostras de Acinetobacter baumannii isoladas de pacientes internados em unidades de terapia intensiva de um hospital terciário

Pós-graduação em Microbiologia - IBILCE

Opportunistic pathogens and elements of the resistome that are common in bottled mineral water support the need for continuous surveillance

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Perfil molecular e resistência a antimicrobianos de Salmonella isolada em linfonodos mesentéricos de suínos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of Mutation and Genetic Background on Drug Resistance in Mycobacterium tuberculosis

Bacterial factors may contribute to the global emergence and spread of drug-resistant tuberculosis (TB). Only a few studies have reported on the interactions between different bacterial factors. We studied drug-resistant Mycobacterium tuberculosis isolates from a nationwide study conducted from 2000 to 2008 in Switzerland. We determined quantitative drug resistance levels of first-line drugs by using Bactec MGIT-960 and drug resistance genotypes by sequencing the hot-spot regions of the relevant genes. We determined recent transmission by molecular methods and collected clinical data. Overall, we analyzed 158 isolates that were resistant to isoniazid, rifampin, or ethambutol, 48 (30.4%) of which were multidrug resistant. Among 154 isoniazid-resistant strains, katG mutations were associated with high-level and inhA promoter mutations with low-level drug resistance. Only katG(S315T) (65.6% of all isoniazid-resistant strains) and inhA promoter -15C/T (22.7%) were found in molecular clusters. M. tuberculosis lineage 2 (includes Beijing genotype) was associated with any drug resistance (adjusted odds ratio [OR], 3.0; 95% confidence interval [CI], 1.7 to 5.6; P < 0.0001). Lineage 1 was associated with inhA promoter -15C/T mutations (OR, 6.4; 95% CI, 2.0 to 20.7; P = 0.002). We found that the genetic strain background influences the level of isoniazid resistance conveyed by particular mutations (interaction tests of drug resistance mutations across all lineages; P < 0.0001). In conclusion, M. tuberculosis drug resistance mutations were associated with various levels of drug resistance and transmission, and M. tuberculosis lineages were associated with particular drug resistance-conferring mutations and phenotypic drug resistance. Our study also supports a role for epistatic interactions between different drug resistance mutations and strain genetic backgrounds in M. tuberculosis drug resistance.