Análise da prevalência de resistência aos antimicrobianos em pacientes com infecção urinária comunitária de um laboratório privado na cidade do Rio de Janeiro

Descrever a prevalência das espécies bacterianas isoladas nas infecções urinárias comunitárias. Descrever os perfis de susceptibilidade aos antibióticos de uso oral utilizado frente às bactérias isoladas nas infecções urinárias comunitárias. Avaliar a prevalência de fenótipos de resistência bacterianos através dos resultados dos testes de susceptibilidade e dos rastreamentos específicos utilizados. Amostras colhidas exclusivamente no atendimento ambulatorial com contagens de unidades formadoras de colônias entre 100.000 a ≥1.000.000 por mililitro (UCF/ml) Com ou sem piúria no exame de elementos anormais na urina e sedimentoscopia (EAS). Foram analisados retrospectivamente os resultados de urinoculturas e dos testes de susceptibilidade a antimicrobianos, realizados em um Laboratório da rede privada na cidade do Rio de janeiro, de pacientes atendidos em ambulatórios e com quadros de ITU. As amostras de urina coletadas englobavam basicamente os seguintes bairros: Botafogo, Barra da Tijuca, Ipanema, Copacabana, Tijuca e Centro. Foram analisados um total de 8.475 culturas de urina divididas em 7.286 urinas de pacientes femininos e 1.189 de pacientes masculinos entre Janeiro de 2006 a Dezembro de 2012. As amostras foram todas coletadas na Cidade do Rio de Janeiro e englobavam basicamente os seguintes bairros: Botafogo, Barra da Tijuca, Ipanema, Copacabana, Tijuca e Centro. Encontramos um percentual de resistência de 27% para ciprofloxacina frente à Escherichia coli que com 68.23% é a principal etiologia encontrada na ITU na comunidade os resultados das três fluoroquinolonas avaliadas no estudo, ciprofloxacina (2 geração), levofloxacina (3 geração) e norfloxacina (2 geração), acharemos respectivamente 27%, 25% e 20% de resistência em Escherichia coli. O uso de fluoroquinolonas em infecções urinárias comunitárias e consequentemente os achados de padrões de resistência neste estudo, reforçam o que já foi descrito em outros trabalhos. A cefalosporina de 2 geração (cefuroxima), demonstrou percentuais de resistência bastante satisfatórios frente as principais etiologias. Em Escherichia coli o percentual foi de 2%, em Klebsiella pneumoniae 3% e em Proteus mirabilis não houve nenhum achado de resistência. Uma das vantagens da cefuroxima é ser ativa quanto à produção de beta lactamase, conferindo um espectro maior frente a possíveis produtoras desta enzima. Seu esquema posológico é de 250mg duas vezes ao dia por 7 dias para infecções urinárias não complicadas. O meio mais eficaz de melhorar a administração antimicrobiana provavelmente envolverá um programa abrangente que incorpora múltiplas estratégias e colaboração entre as diversas especialidades dentro de uma determinada instituição de saúde. Neste contexto, a observação periódica da incidência bacteriana com seus respectivos índices de resistência aos antimicrobianos por sitio de infecção e correlação com os antibióticos mais comumente utilizados, é mandatória para o sucesso terapêutico.

Structure and function of sedoheptulose-7-phosphate isomerase, a critical enzyme for lipopolysaccharide biosynthesis and a target for antibiotic adjuvants

The barrier imposed by lipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria presents a significant challenge in treatment of these organisms with otherwise effective hydrophobic antibiotics. The absence of L-glycero-D-manno-heptose in the LPS molecule is associated with a dramatically increased bacterial susceptibility to hydrophobic antibiotics and thus enzymes in the ADP-heptose biosynthesis pathway are of significant interest. GmhA catalyzes the isomerization of D-sedoheptulose 7-phosphate into D-glycero-D-manno-heptose 7-phosphate, the first committed step in the formation of ADP-heptose. Here we report structures of GmhA from Escherichia coli and Pseudomonas aeruginosa in apo, substrate, and product-bound forms, which together suggest that GmhA adopts two distinct conformations during isomerization through reorganization of quaternary structure. Biochemical characterization of GmhA mutants, combined with in vivo analysis of LPS biosynthesis and novobiocin susceptibility, identifies key catalytic residues. We postulate GmhA acts through an enediol-intermediate isomerase mechanism.

Diphenylamine increases cloacin DF13 sensitivity in avian septicemic strains of Escherichia coli

Thirteen avian septicemic isolates of Escherichia coli were examined for the presence of the aerobactin iron transport system. All of the strains possessed a functional aerobactin system and hybridization experiments showed that the aerobactin genes were located on ColV-type plasmids in all cases. The expression of the aerobactin receptor IutA was also studied by determining the bacterial susceptibility to the bacteriocin cloacin DF13. Twelve of the 13 isolates were cloacin-resistant but became sensitive to this bacteriocin upon treatment with diphenylamine which caused a reduction in the amount of O-side chain lipopolysaccharide.

Putrescine Reduces Antibiotic-Induced Oxidative Stress as a Mechanism of Modulation of Antibiotic Resistance in Burkholderia cenocepacia

Communication of antibiotic resistance among bacteria via small molecules is implicated in transient reduction of bacterial susceptibility to antibiotics, which could lead to therapeutic failures aggravating the problem of antibiotic resistance. Released putrescine from the extremely antibiotic resistant bacterium Burkholderia cenocepacia protects less resistant cells from different species against the antimicrobial peptide polymyxin B (PmB). Exposure of B. cenocepacia to sub-lethal concentrations of PmB and other bactericidal antibiotics induce reactive oxygen species (ROS) production and expression of the oxidative stress response regulator OxyR. We evaluated whether putrescine alleviates antibiotic-induced oxidative stress. The accumulation of intracellular ROS such as superoxide ion and hydrogen peroxide was assessed fluorometrically with dichlorofluorescein diacetate, while the expression of OxyR and putrescine synthesis enzymes was determined in luciferase assays using chromosomal promoter-lux reporter system fusions. We evaluated wild type and isogenic deletion mutant strains with defects in putrescine biosynthesis after exposure to sub-lethal concentrations of PmB and other bactericidal antibiotics. Exogenous putrescine protected against oxidative stress induced by PmB and other antibiotics, whereas reduced putrescine synthesis resulted in increased ROS generation, and a parallel increased sensitivity to PmB. Of the 3 B. cenocepacia putrescine synthesizing enzymes, PmB induced only BCAL2641, an ornithine decarboxylase. This study exposes BCAL2641 as a critical component of the putrescine-mediated communication of antibiotic resistance, and as a plausible target for designing inhibitors that would block the communication of such resistance among different bacteria, ultimately reducing the window of therapeutic failure in treating bacterial infections.

Perfil microbiológico, prevalencia de resistencia antibiótica y patrones de susceptibilidad en infección urinaria en una población pediátrica

Introducción: El incremento de la resistencia antibiótica se considera un problema de salud pública con consecuencias clínicas y económicas, por lo tanto se determinará la prevalencia de resistencia antibiótica en Infección del Tracto Urinario (ITU, el perfil microbiológico y los patrones de susceptibilidad en una población pediátrica atendida en la Fundación Cardioinfantil. Materiales y métodos: Estudio observacional de corte transversal, retrospectivo, entre 1 mes a 18 años de edad, con diagnóstico de ITU comunitaria atendidos entre Enero de 2011 y Diciembre de 2013. Se excluyeron pacientes con dispositivos en la vía urinaria, instrumentación quirúrgica previa, trayectos fistulosos entre la vía urinaria y sistema digestivo, ITU luego de 48 horas de hospitalización y recaída clínica en tratamiento. Se estableció la prevalencia de ITU resistente y se realizó un análisis descriptivo de la información. Resultados: Se evaluaron 385 registros clínicos, con una mediana de 1.08 años (RIQ 0.8 – 4.08), el 73.5% eran niñas. La fiebre predominó (76.5%), seguido de emesis (32.0%), disuria (23.7%) y dolor abdominal (23.1%). El uropatógeno más frecuente fue E.coli (75%), seguido de Proteus mirabilis (8.5%) y Klebsiella spp. (8.3%). La Ampicilina, el Trimetropim sulfametoxazol, la Ampicilina sulbactam y el ácido nalidixico tuvieron mayor tasa de resistencia. La prevalencia de BLEE fue 5.2% y AmpC 3.9%. La prevalencia de resistencia antimicrobiana fue de 11.9%. Conclusiones: La E.coli es el uropatogeno más frecuentemente aislado en ITU, con resistencia a la ampicilina en 60.2%, cefalosporinas de primera generación en 15.5%, trimetropin sulfametoxazol en 43.9%, cefepime 4.8%. La prevalencia de resistencia antimicrobiana fue de 11.9%.

Microbiological evaluation of different strategies for management of snakes in captivity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Atividade comparada da cefepima, cefpiroma e amicacina em amostras bacterianas hospitalares

The objective of this study was to evaluate the in vitro activity of cefepime, cefpirome and amikacin against the most prevalent nosocomial bacteria. Initially a prospective study was designed to compare the bacterial susceptibility to the three drugs using 1,022 pathogenic strains. The strains were isolated from hospitalized patients of the Hospital das Clinicas - Faculdade de Medicina de Botucatu, SP, from March to December of 1996, by using the Bauer-Kirby susceptibility diffusion controlled method. The activity of cefepime by the Kirby-Bauer method was significantly higher (χ2, p ≤ 0.05) than cefpirome and amikacin for the following bacteria: P. aeruginosa (72% x 56% x 64%, respectively), Enterobacter cloacae (98% x 88% x 80%) and total strains (79.5% x 74.3% x 76.8%). Cefpirome exhibited higher activity than cefepime only to Enterococcus faecalis (42% x 23%). In the 12 other bacterial groups studied the sensibility of the three drugs was similar (χ2, p ≥ 0.05). The minimal inhibitory concentration (MIC) for 127 bacterial strains - Enterobacter cloacae (12), Citrobacter sp (15), Pseudomonas aeruginosa (50), Acinetobacter baumannii (12), BGNF others (22) and Enterococcus faecalis (16)-from the same origin previously described and isolated during 1997, was determined by E-test. Ranges of MIC intervals, MIC(50%), MIC(90%) and the proportion of the sensitive bacterial strains were determined and permitted the following analysis: the activity of cefepime against Gram-negative bacteria was 2 or more times higher than that of cefpirome and amikacin, specially when CIM(90%) was considered; the activity of cefpirome was higher only against E. faecalis. This information must be considered in the rational use of antibiotic, specially in patients with nosocomial infections.

Peptídeos derivados da proteína bacteriana YacG : síntese e estudos de estrutura-função

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência de aplicações do laser érbio:YAG sobre a viabilidade microbiana, sua resistência a drogas e atividade hemolítica

Pós-graduação em Odontologia - FOA

Effect of sub-lethal challenge with Photodynamic Antimicrobial Chemotherapy (PACT) on the antibiotic susceptibility of clinical bacterial isolates

This study aimed to determine the effect of sub-lethal challenge with Photodynamic Antimicrobial Chemotherapy (PACT) on the susceptibility of clinical Staphylococcus aureus and Pseudomonas aeruginosa isolates to both PACT and a range of antibiotics used in the treatment of infection caused by these bacteria. Clinical S. aureus and P. aeruginosa isolates were exposed to sub-lethal PACT with meso-tetra (N-methyl-4-pyridyl) porphine tetra tosylate (TMP) and methylene blue (MB) over a 72 h period. After exposure, susceptibility of surviving organisms to a range of antibiotics was determined and compared with the susceptibility of an untreated control. Surviving bacteria were also exposed to previously lethal photosensitizer-light combinations, to determine if susceptibility to PACT was affected by sub-lethal exposure. Exposure to sub-lethal PACT did not decrease susceptibility to antibiotics with the minimum inhibitory concentrations for 95% and 100% of P. aeruginosa and S. aureus isolates, respectively, within two doubling dilutions of the MIC of the untreated control. Similarly, habituation with sub-lethal PACT did not reduce susceptibility of P. aeruginosa isolates to PACT levels previously determined as lethal. A reduction in susceptibility to PACT following habituation was apparent for two S. aureus isolates with MB and for 1 S. aureus isolate with IMP. However, for two of these three isolates, the log reduction for habituated cells was still greater than 4 log(10). PACT remains an attractive potential treatment for infection caused by these bacteria. (C) 2010 Elsevier B.V. All rights reserved.

Susceptibility of different insect pupae to the bacterial symbiont, Xenorhabdus nematophila, isolated from the entomopathogenic nematode Steinernema carpocapsae

Cells of the bacterial symbiont Xenorhabdus nematophila from the entomopathogenic nematode, Steinernema carpocapsae entered the pupae of Plutella xylostella after 15 minutes treatment with suspensions containing the bacterial cells. Secretions of Xenorhabdus nematophila, in either broth or water, were found lethal to the pupae of P. xylostella when applied in moist sand. The bacterial symbiont Xenorhabdus nematophila was found lethal to the pupae of greater wax moth (Galleria mellonella), beet armyworm (Spodoptera exigua), diamondback moth (Plutella xylostella) and black vine weevil (Otiorhynchus sulcatus) in the absence of the nematode vector and the cells of X. nematophila entered the haemocoele of the pupae.

Analysis of susceptibility profile of Pseudomonas spp. and prevalence of bacterial samples from the surfaces of dental consulting-rooms

The aim of this research was to evaluate the susceptibility profile of Pseudomonas spp. and the prevalence of bacterial samples isolated from horizontal surfaces surrounding wash-basins used by dentists in several adjoined consulting-rooms, at points next to and at a distance from the basin, before and after surgical procedures. Our results showed a high percentage of Gram-positive cocci and Gram-negative bacilli; 34.66% were Staphylococcus spp. and 30.12% were non-fermentative Gram-negative bacilli among which Pseudomonas spp. (40.90%) was the commonest genus. Analysis of the susceptibility profile of Pseudomonas spp. isolates by determining the minimal inhibitory concentration (MIC) of 14 antibiotics showed a great variation among the strains and high rates of resistance to cefazolin, ceftazidime and aztreonan. Of the 14 antibiotics tested, 59.03% were found to be active against all the environmental isolates. Strains were resistant to aztreonan (62.82%), while susceptibility to third generation cephalosporins was variable.

Exogenous bacterial osteomyelitis in 52 dogs: a retrospective study of etiology and in vitro antimicrobial susceptibility profile (2000-2013)

Background: Most clinical cases of osteomyelitis in dogs involve infectious agents, especially bacteria and fungi. The characterization of these microorganisms may aid in the prevention and treatment of disease.Objective: The aim of this study was to evaluate retrospectively microbiological cultures and in vitro antimicrobial susceptibility profile of isolates from 52 cases of bacterial osteomyelitis in long bones of dogs over 2000-2013. In 78% of the cases injuries were caused by a motor vehicle accident, but there were a few cases of dog bites (17%) and ascending infection due to pododermatitis (5%).Animals and methods: The isolated microorganisms were identified based on conventional phenotypic methods. In vitro disk diffusion test was performed using 30 different antimicrobials.Results: The isolates were obtained from femur (28%), humerus (16%), tibia (31%), and radius/ulna (25%). Among 52 cases, culture was positive in 88% of cases. Thirteen genus of different species of microorganisms were isolated. The most common microorganisms isolated were Staphylococcus spp. and Escherichia coli followed by Streptococcus spp., enteric bacteria, Corynebacterium sp. and anaerobic bacteria. In 42% of cases cultures were mixed. The most effective drugs against isolated bacteria were amoxicillin and clavulanate potassium (79%) followed by ceftriaxone (69%). High-resistance rates were documented against azithromycin (80%), penicillin (59%), and clindamycin (59%).Conclusions: The present study highlights diverse etiologic agents in cases of infectious bacterial osteomyelitis, with predominance of Staphylococcus genus, and reinforces the importance of obtaining cultures and susceptibility profiles given the high rates of antimicrobial resistance.