Exopolysaccharides from Klebsiella oxytoca: anti-inflammatory activity

Abstract Exopolysaccharides (EPS) produced by Klebsiella oxytoca are of environmental, pharmaceutical, and medicinal interest. However, studies about the anti-inflammatory activity of EPS produced by this microorganism still remain limited. The aim of this study was to produce, characterize, and evaluate the anti-inflammatory activity of EPS from K. oxytoca in a pleurisy model. Colorimetric analysis revealed that precipitated crude exopolysaccharides (KEPSC) and deproteinated exopolysaccharides (KEPS) present high levels of total carbohydrates (65.57% and 62.82%, respectively). Analyses of uronic acid (7.90% in KEPSC and 6.21% in KEPS) and pyruvic acid (3.01% in KEPSC and 1.68% in KEPS) confirm that the EPS are acidic. Gas chromatography-mass spectrometry analyses demonstrated that the EPS consisted of rhamnose (29.83%), glucose (11.21%), galactose (52.45%), and mannose (6.50%). The treatment of an experimental pleurisy model in rats through subcutaneous administration of 50, 100, 200, and 400 mg/kg of KEPS decreased both the volume of inflammatory exudate and the number of leukocytes recruited to the pleural cavity. The present data showed that EPS production by K. oxytoca using the method described is easy to perform and results in a good yield. In addition, we show that KEPS exhibit anti-inflammatory activity when administered subcutaneously in rats.

Cord factor producer Mycobacterium abscessus subsp. bolletii in asymptomatic immunocompetent host sputa samples

Abstract We report a case of Mycobacterium abscessus subsp. bolletii colonization in upper respiratory tract of an immunocompetent patient, who was misdiagnosed as tuberculosis by Acid Fast Bacilli (AFB) and cord factor formation observed directly from the sputa culture in liquid medium. This fact reflected a significant impact on the individual case's life and showed the importance to identify the mycobacteria isolated from clinical sample at species level, and to determine the true implication of nontuberculous mycobacteria (NTM) detected in clinical samples.

Occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens

Mycoplasmaspp. and Ureaplasmaspp. belong tohumans'genitourinary microbiota and sometimesare associated with infections of the genitourinarytract. The aim of this study was to evaluate the occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens from patients of the 15thRegional de Saúde of ParanáState, Brazil, and to correlate the results with clinical and laboratory data.A retrospective cross-sectional study was conducted,based on the analysis of results of vaginal, endocervical, urine andurethral culture for mycoplasmas from patients attended in areference laboratory, from January 2009 to December 2016. We evaluated 2,475 results of culture for mycoplasmas. A total of 50.8% patients were positive for mycoplasmas. Of these, 76.8%had positive culture exclusively for Ureaplasmaspp. and 4.7% for Mycoplasmahominis. Both microorganisms were isolated in the microbiology culture of 18.5% of patients. Among the positive culture, 81.4% had significant concentrations.Bacterialvaginosis was the most common alteration observed in association with mycoplasmas.Thehigh positivity of cultures for mycoplasmas, especially Ureaplasmaspp. found in our study, highlightthe presence of these microorganisms in many of the genital tract disorders that can be sexually transmitted and, consequently, should not be neglected.

Is the efflux pump inhibitor Verapamil a potential booster for isoniazid against Mycobacterium tuberculosis?

The membrane-based efflux pump systems are recognized to have an important role in pathogenicity and drug resistance in Mycobacterium tuberculosis by the extrusion of toxic substrates and drugs from the inner bacillus. This study aimed to investigate the in vitro interaction of Verapamil (VP), an efflux pump inhibitor, with the classical first-line anti-tuberculosis drug isoniazid (INH) in resistant and susceptible M. tuberculosis clinical isolates. Seven multidrug-resistant (MDR), three INH monoresistant and four susceptible M. tuberculosis clinical isolates were tested for the INH and VP combination by modified Resazurin Microtiter Assay Plate (REMA). Fractional Inhibitory Concentration (FIC) and Modulation Factor (MF) were determined. The INH plus VP combination showed no significant change in the Minimum inhibitory concentration (MIC) values of INH (FIC≥ 0.5; MF=1 or 2).The use of VP in tuberculosis therapy should be managed carefully, considering the resistance caused by specific mutation in katG and inhA genes, in which the use of these EPIs may have no success. The use of EPIs as an adjunctive drug in the anti-tuberculosis therapy should be further investigated on a larger number of M. tuberculosis clinical isolates with different resistant profile.

Fast detection of Mycobacterium tuberculosis in culture-positive sputum samples by nitrate reductase activity

ABSTRACT Microscopy and bacterial culture are the main tools in the diagnosis of tuberculosis. Since the slow growth of Mycobacterium tuberculosis impairs rapid diagnosis strategies, especially in countries where the latter are the only available resources, the ongoing development of new and inexpensive tools based on mycobacterial metabolism optimizing growth detection with preliminary identification is greatly welcome. When compared to the other species from the M. tuberculosis complex, M. tuberculosis is a strong nitrate reducer. Current assay compares the nitrate reductase activity of M. tuberculosis from pulmonary specimens cultivated in nitrate-supplemented media. Fifty-five sputum samples were decontaminated and inoculated in conventional (Middlebrook 7H9, Ogawa Kudoh-OK) and in nitrate-supplemented media (Middlebrook 7H9-N, Ogawa Kudoh-N). An aliquot from the media directly reacted with Griess reagent (7H9-N and OK-N) every five days, or transferred to a nitrate substrate solution (7H9, OK). Nitrate to nitrite reduction was considered positive, revealed by the pink color, indicating bacterial growth. As reference method, the Mycobacteria Growth Indicator Tube (MGIT) was used for sensitivity calculations and statistical analysis. 7H9-N and OK-N assays proved to perform better in detecting M. tuberculosis than conventional assays (7H9 and OK). Indeed, broth nitrate-supplemented medium (7H9-N) was comparable to MGIT to detect M. tuberculosis, except in growth detection time. Results show that 7H9-N may be used as an alternative tool particularly in low-income countries since it is a simple and cheap technique, and does not restrict diagnosis to single-source products.

Molecular characterization of Mycobacterium tuberculosis and Mycobacterium bovis isolates by Enterobacterial Repetitive Intergenic Consensus-PCR

Tuberculosis (TB) is an infectious disease in which the molecular typing methods allow to have important information about the dynamics of transmission and to assist properly in disease control. Although the ERIC-PCR (Enterobacterial repetitive intergenic consensus-PCR) assay is fast and easy to perform, scarce studies have reported its use in epidemiological studies in TB outbreaks. In this study, we aimed to genotype Mycobacterium tuberculosis and M. bovis isolates by ERIC-PCR and compare its discriminatory power with two other classically used methods: 12 loci-MIRU (Mycobacterial Interspersed Repetitive Units) and Spoligotyping. The M. tuberculosis isolates studied were from northwestern and southwestern and M. bovis from northwestern Parana, Brazil. ERIC-PCR rendered banding patterns with great diversity (1 to 12 bands) of molecular sizes, ranging from 100 to 1600 bp. ERIC-PCR showed to be fast, simple and affordable to differentiate isolates. ERIC-PCR would be an important tool in the epidemiology of TB as screening in case of outbreak, which demands rapid intervention. However if any doubt persist, as it may occur with the application of only one genotypic method, other genotyping methods should be applied and carefully interpreted, always with additional epidemiological information.

ß-lactamase-producing Gram-negative bacteria in an intensive care unit in southern Brazil

ABSTRACT The present study evaluated the antimicrobial susceptibility profile, ß-lactamase production, and genetic diversity of Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter spp. using phenotypic identification, antimicrobial susceptibility testing, and ß-lactamase phenotypic detection. Isolates were obtained from patients in an intensive care unit in a hospital in southern Brazil. Bacterial genomic DNA was extracted, followed by the genotypic detection of carbapenemases and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). Fifty-six isolates (26 Klebsiella pneumoniae, five Escherichia coli, three Enterobacter aerogenes, nine P. aeruginosa, and 13 Acinetobacter spp.) were evaluated. The phenotypic extended spectrum ß-lactamase (ESBL) test was positive in 53.8% of the K. pneumoniae isolates, 100.0% of the E. coli isolates, and 100.0% of the E. aerogenes isolates. Phenotypic and genotypic testing of K. pneumoniae carbapenemase (KPC) was positive in 50.0% of the K. pneumoniae isolates. Phenotypic and genotypic testing showed that none of the P. aeruginosa or Acinetobacter spp. isolates were positive for metallo- ß-lactamase (MBL). The bla OXA gene was detected only in Acinetobacter spp. The lowest genetic diversity, determined by ERIC-PCR, was observed among the KPC-producing K. pneumoniae isolates and OXA-producing Acinetobacter spp. isolates, indicating the inadequate dissemination control of multidrug-resistant bacteria in this hospital environment.

Critical analysis: use of polymerase chain reaction to diagnose leprosy

ABSTRACT Leprosy is a neglected tropical disease and an important public health problem, especially in developing countries. It is a chronic infectious disease that is caused by Mycobacterium leprae, which has a predilection for the skin and peripheral nerves. Although it has low sensitivity, slit-skin smear (SSS) remains the conventional auxiliary laboratory technique for the clinical diagnosis of leprosy. Polymerase chain reaction (PCR) is a molecular biology technique that holds promise as a simple and sensitive diagnostic tool. In the present study, the performance of two PCR methods, using different targets, PCR-LP and PCR-P, were compared with SSS with regard to leprosy diagnosis in a reference laboratory. M. leprae DNA was extracted from 106 lymph samples of 40 patients who had clinical suspicion of leprosy. The samples were subjected to both PCR techniques and SSS. Amplification of the human b-globin gene was used as PCR inhibitor control. The specificity of both PCR techniques was 100%, and sensitivity was 0.007 and 0.015 µg/ml for PCR-LP and PCR-P, respectively. No significant difference was found between either the PCR-LP or PCR-P results and SSS results (p > 0.05). Although PCR is not yet a replacement for SSS in the diagnosis of leprosy, this technique may be used as an efficient auxiliary tool for early detection of the disease, especially in endemic regions. This strategy may also be useful in cases in which SSS results are negative (e.g., in paucibacillary patients) and cases in which skin biopsy cannot be performed.

RESUMO A hanseníase é uma doença tropical negligenciada e ainda um importante problema de saúde pública, especialmente nos países em desenvolvimento. É uma doença infecciosa crônica causada pelo Mycobacterium leprae, que tem predileção pela pele e nervos periféricos. Embora com baixa sensibilidade, o esfregaço de linfa (SSS) continua sendo o método laboratorial convencional auxiliar no diagnóstico clínico da hanseníase. A biologia molecular representada pela Reação em Cadeia da Polimerase (PCR) trouxe a expectativa de ser uma ferramenta diagnóstica simples e sensível. No presente estudo, o desempenho de dois métodos de PCR usando alvos diferentes, PCR-P e PCR-LP, foi comparado com SSS no diagnóstico da hanseníase em um laboratório de referência. DNA de M. leprae foi extraído de 106 amostras de linfa de 40 pacientes que apresentavam suspeita clínica de hanseníase. As amostras foram submetidas tanto a PCR como SSS. A amplificação do gene humano β-globina foi usada como controle de inibição da PCR. A especificidade de ambas as técnicas de PCR foi de 100% e a sensibilidade foi de 0,007 μg/mL e 0,015 μg/mL para a PCR-P e PCR-LP, respectivamente. Não se observou diferença estatística entre os resultados da PCR-LP e PCR-P, quando comparado com SSS (p > 0,05). Apesar de a PCR ainda não substituir o SSS no diagnóstico da hanseníase, esta técnica pode ser usada como ferramenta auxiliar eficiente para a detecção precoce da doença, especialmente em regiões endêmicas. Esta estratégia pode também ser útil nos casos em que os resultados de SSS forem negativos (ex. em pacientes paucibacilares) e em casos onde a biópsia da pele não pode ser realizada.

Occurrence of Mycobacterium bovis and non-tuberculous mycobacteria (NTM) in raw and pasteurized milk in the northwestern region of Paraná, Brazil

Milk is widely consumed in Brazil and can be the vehicle of agent transmission. In this study, was evaluated the occurrence of Mycobacterium bovis and non-tuberculous mycobacteria (NTM) in raw and pasteurized milk consumed in the northwestern region of Paraná, Brazil. Fifty-two milk samples (20 pasteurized and 32 raw) from dairy farms near the municipality of Maringa, Parana State, Brazil were collected. Milk samples were decontaminated using 5% oxalic acid method and cultured on Lowenstein-Jensen and Stonebrink media at 35 °C and 30 °C, with and without 5-10% CO2. Mycobacteria isolates were identified by morphological features, PCR-Restriction Fragment Length Polymorphism Analysis (PCR-PRA) and Mycolic acids analysis. Thirteen (25%) raw and 2 (4%) pasteurized milk samples were positive for acid fast bacilli growth. Nine different species of NTM were isolated (M. nonchromogenicum, M. peregrinum, M. smegmatis, M. neoaurum, M. fortuitum, M. chelonae, M. flavescens, M. kansasii and M. scrofulaceum). M. bovis was not detected. Raw and pasteurized milk may be considered one source for NTM human infection. The paper reinforces the need for intensification of measures in order to avoid the milk contamination and consequently prevent diseases in the south of Brazil.

Anti-Mycobacterium tuberculosis activity and cytotoxicity of Calophyllum brasiliense Cambess (Clusiaceae)

We evaluated the in vitro anti-Mycobacterium tuberculosis activity and the cytotoxicity of dichloromethane extract and pure compounds from the leaves of Calophyllum brasiliense. Purification of the dichloromethane extract yielded the pure compounds (-) mammea A/BB (1), (-) mammea B/BB (2) and amentoflavone (3). The compound structures were elucidated on the basis of spectroscopic and spectrometric data. The contents of bioactive compounds in the extracts were quantified using high performance liquid chromatography coupled to an ultraviolet detector. The anti-M. tuberculosis activity of the extracts and the pure compounds was evaluated using a resazurin microtitre assay plate. The cytotoxicity assay was performed in J774G.8 macrophages using the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide colourimetric method. The quantification of the dichloromethane extract showed (1) and (2) at concentrations of 31.86 ± 2.6 and 8.24 ± 1.1 µg/mg of extract, respectively. The dichloromethane and aqueous extracts showed anti-M. tuberculosis H37Rv activity of 62.5 and 125 µg/mL, respectively. Coumarins (1) and (2) showed minimal inhibitory concentration ranges of 31.2 and 62.5 µg/mL against M. tuberculosis H37Rv and clinical isolates. Compound (3) showed no activity against M. tuberculosis H37Rv. The selectivity index ranged from 0.59-1.06. We report the activity of the extracts and coumarins from the leaves of C. brasiliense against M. tuberculosis.

COMPARISON BETWEEN AUTOMATED SYSTEM AND PCR-BASED METHOD FOR IDENTIFICATION AND ANTIMICROBIAL SUSCEPTIBILITY PROFILE OF CLINICAL Enterococcus spp / Comparação entre o sistema automatizado e PCR na identificação e susceptibilidade de isolados clínicos de Enterococcus spp

Enterococci are increasingly responsible for nosocomial infections worldwide. This study was undertaken to compare the identification and susceptibility profile using an automated MicrosScan system, PCR-based assay and disk diffusion assay of Enterococcus spp. We evaluated 30 clinical isolates of Enterococcus spp. Isolates were identified by MicrosScan system and PCR-based assay. The detection of antibiotic resistance genes (vancomycin, gentamicin, tetracycline and erythromycin) was also determined by PCR. Antimicrobial susceptibilities to vancomycin (30 µg), gentamicin (120 µg), tetracycline (30 µg) and erythromycin (15 µg) were tested by the automated system and disk diffusion method, and were interpreted according to the criteria recommended in CLSI guidelines. Concerning Enterococcus identification the general agreement between data obtained by the PCR method and by the automatic system was 90.0% (27/30). For all isolates of E. faecium and E. faecalis we observed 100% agreement. Resistance frequencies were higher in E. faecium than E. faecalis. The resistance rates obtained were higher for erythromycin (86.7%), vancomycin (80.0%), tetracycline (43.35) and gentamicin (33.3%). The correlation between disk diffusion and automation revealed an agreement for the majority of the antibiotics with category agreement rates of > 80%. The PCR-based assay, the van(A) gene was detected in 100% of vancomycin resistant enterococci. This assay is simple to conduct and reliable in the identification of clinically relevant enterococci. The data obtained reinforced the need for an improvement of the automated system to identify some enterococci.

Os enterococos são cada vez mais responsáveis por infecções hospitalares em todo o mundo. Este estudo foi realizado para comparar a identificação e perfil de suscetibilidade entre o sistema automatizado MicrosScan e a técnica molecular de PCR em espécies de Enterococcus spp. Foram avaliados 30 isolados clínicos de Enterococcus spp. Os isolados foram identificados pelo sistema MicrosScan® e pela técnica de PCR. A detecção de genes de resistência a antibióticos (vancomicina, gentamicina, tetraciclina e eritromicina) foi determinada por PCR. Suscetibilidades antimicrobianas à vancomicina (30 µg), gentamicina (120 µg), tetraciclina (30 µg) e eritromicina (15 µg), foram testados pelos métodos automatizados e pelo disco difusão, de acordo com as orientações do CLSI. No que diz respeito à identificação de Enterococcus em geral entre os dados obtidos pelo método de PCR e pelo sistema automático foi de 90,0% (27/30). Para todos os isolados de E. faecium e E. faecalis observamos concordância de 100%. Freqüências de resistência foi maior em E. faecium do que em E. faecalis. As taxas de resistência obtidas foi maior para eritromicina (86,7%), vancomicina (80,0%), tetraciclina (43,35%) e gentamicina (33,3%). A correlação entre a técnica de disco difusão e automação revelou-se de acordo para maioria dos antibióticos com taxas > 80%. O gene van(A) foi detectado em 100% dos Enterococcus resistentes á vancomicina. O ensaio baseado em PCR é de simples realização e de confiança para identificação de enterococos clinicamente relevantes. Os dados obtidos reforçam a necessidade de melhoria no sistema automatizado para identificar alguns enterococos.

High genetic diversity among Pseudomonas aeruginosa and Acinetobacter spp. isolated in a public hospital in Brazil

In Brazil and other regions of the world, Pseudomonas aeruginosa and Acinetobacter spp. have emerged as important agents of nosocomial infection and are commonly involved in outbreaks. The main objective of the present study was to evaluate the genetic relationship among P. aeruginosa and Acinetobacter spp. isolated from patients in a public university hospital in northwestern Paraná, Brazil, and report their antimicrobial resistance profile. A total of 75 P. aeruginosa and 94 Acinetobacter spp. isolates were phenotypically identified and tested for antibiotic susceptibility using automated methodology. Polymyxin B was tested by disk diffusion for P. aeruginosa. Metallo-β-lactamase (MBL) was detected using a disk approximation test. Genotyping was performed using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR). Approximately 55% of the P. aeruginosa isolates and 92% of the Acinetobacter spp. isolates were multiresistant, but none were MBL-producers. ERIC-PCR revealed the presence of small clusters of carbapenem-resistant Acinetobacter spp., most likely OXA-type carbapenemase producers. Furthermore, high genetic diversity in P. aeruginosa and Acinetobacter spp. clinical isolates was observed, suggesting that cross-transmission is not very frequent in the studied hospital.

No Brasil, bem como em outras regiões do mundo, Pseudomonas aeruginosa e Acinetobacter spp. surgiram como importantes agentes de infecção nosocomial e são comumente envolvidos em surtos. O objetivo principal deste estudo foi descrever a relação genética de P. aeruginosa e Acinetobacter spp. isoladas de pacientes internados em hospital universitário público do noroeste do Paraná - Brasil e reportar o perfil de resistência dessas bactérias. Um total de 75 P. aeruginosa e 94 Acinetobacter spp. isolados foi fenotipicamente identificado e testado para a suscetibilidade aos antibióticos por metodologia automatizada. A polimixina B foi testada por difusão em disco para P. aeruginosa. Metalo-β-lactamase (MBL) foi detectada por disco-aproximação. Análise genotípica foi realizada por enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR). Aproximadamente 55% dos isolados de P. aeruginosa e 92% de Acinetobacter spp. isolados foram multirresistentes, mas nenhum foi produtor de MBL. Os resultados de ERIC-PCR revelaram pequenos grupamentos de Acinetobacter spp. resistentes aos carbapenêmicos, provavelmente pela produção de carbapenemases do tipo OXA. Além disso, alta diversidade genética entre os isolados de P. aeruginosa e Acinetobacter spp. foi observada, sugerindo que a transmissão cruzada destas espécies bacterianas não é muito frequente em nosso hospital.

Use of the Ogawa-Kudoh method to isolate mycobacteria in a tuberculosis reference laboratory in northwestern Paraná, Brazil

Culturing is the gold standard method for confirming a diagnosis of tuberculosis (TB). The Brazilian Ministry of Health recently proposed the use of the Ogawa-Kudoh method for sputa cultures to detect Mycobacterium tuberculosis. The aim of the present study was to evaluate 8 years of using the Ogawa-Kudoh method in a TB reference laboratory in northwestern Paraná, Brazil. The present study consisted of a retrospective analysis of sputa cultures records for the detection of mycobacteria using the Ogawa-Kudoh method in the Laboratory of Medical Bacteriology, Laboratory of Teaching and Research in Clinical Analysis (LEPAC), State University of Maringá, from July 2003 to September 2011. The following variables were analyzed: Ziehl Neelsen (Z-N) smears and cultures results and the age and gender of the patients. Sputa samples from 3,231 patients with suspected TB were analyzed. Of these, 67.17% were male with an average age of 45.58 years. Of the total number of Z-N-negative samples (n=2,949), 42 (1.42%) were positive for M. tuberculosis (p >0.05). The Ogawa-Kudoh method is an excellent tool for diagnosing pulmonary TB. It is easy to perform, requires less biosafety equipment than the Petroff method, has a low cost, and has good sensitivity for detecting of M. tuberculosis.

A cultura é o método padrão ouro para confirmação da tuberculose (TB). O Ministério da Saúde Brasileiro propôs, recentemente, a utilização do método de Ogawa-Kudoh para cultura de escarro na detecção de Mycobacterium tuberculosis. O objetivo deste estudo foi avaliar oito anos de utilização do método de Ogawa-Kudoh na rotina de um laboratório de referência na região noroeste do Paraná, Brasil. Realizou-se estudo retrospectivo dos registros das culturas de escarro para a detecção de micobactérias, usando o método Ogawa-Kudoh conduzido no Laboratório de Bacteriologia Médica, Laboratório de ensino e pesquisa em Análises Clínicas (LEPAC) da Universidade Estadual de Maringá (UEM), de Julho de 2003 a Setembro de 2011. As seguintes variáveis foram analisadas: esfregaço Ziehl Neelsen (Z-N), cultura, idade e sexo do paciente. Analisaram-se 3.231 amostras de escarro de pacientes com suspeita de tuberculose. Destes, 67,17% eram do sexo masculino com idade média de 45,58 anos. Do total de amostras Z-N negativas (n=2.949), 42 amostras (42/2949, 1,42%) apresentaram cultura positiva para M. tuberculosis (p>0,05). A utilização do método Ogawa-Kudoh representa excelente ferramenta para o diagnóstico precoce da TB pulmonar. É de fácil execução, requer menos equipamentos de biossegurança do que o método de Petroff, apresenta baixo custo e boa sensibilidade para detecção de M. tuberculosis.

High prevalence rate of extended-spectrum beta-lactamases (ESBL) among Enterobacteriaceae in a small Brazilian public hospital

The production of extended-spectrum beta-lactamases (ESBL) is considered one of the most important resistance mechanisms that impair antimicrobial treatment of infections caused by Enterobacteriaceae. Data on culture and susceptibility tests were collected from the Clinical Analyses and Research Laboratory charts reporting on patients admitted to the University Hospital of Maringá (HUM) from January 2004 to December 2009. The following Enterobacteriaceae were selected: Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter spp. and Proteus mirabilis. All tests were performed according to the recommendations of the Clinical and Laboratory Standards Institute (biochemical identification; susceptibility tests; initial screening and phenotypic confirmatory tests for ESBL). For Enterobacter spp. isolates, a disk approximation test was carried out, adding a cefepime disk. Seven hundred samples were analyzed, and E. coli was the most prevalent bacteria (n= 356). ESBLs were detected phenotypically in 7.3 percent of E. coli, 61.7 percent of K. pneumoniae, 33.3 percent of K. oxytoca, 7.1 percent of P. mirabilis, and 13.4 percent of Enterobacter spp samples. Overall ESBL prevalence reached 22 percent when all producers were taken together. Although HUM is considered a small-sized hospital, it showed high levels of resistance to antimicrobial agents, similar to those observed in bigger hospitals, which demonstrated the need for careful epidemiological surveillance.

A produção de beta-lactamases de espectro ampliado (ESBL) é considerada um dos mais importantes mecanismos de resistência aos antimicrobianos, o que dificulta o tratamento de infecções causadas por enterobactérias. Dados sobre cultura e testes de sensibilidade foram coletados das fichas do Laboratório de Ensino e Pesquisa de Análises Clínicas de pacientes atendidos no Hospital Universitário de Maringá (HUM), de janeiro de 2004 a dezembro de 2009. As enterobactérias escolhidas foram: Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter spp. e Proteus mirabilis. Os testes foram realizados de acordo com as recomendações do CLSI (identificação bioquímica; testes de suscetibilidade; triagem e confirmação fenotípica de produção de ESBL). Para isolados de Enterobacter spp., utilizou-se a técnica de disco aproximação, acrescentando um disco de cefepima. Setecentas amostras foram analisadas e E. coli foi a bactéria mais prevalente (n=356). ESBLs foram detectadas fenotipicamente em 7,3 por cento das amostras de E. coli, 61,7 por cento das de K. pneumoniae, 3,3 por cento das de K. oxytoca, 7,1 por cento das de P. mirabilis e em 13,4 por cento das de Enterobacter spp. A prevalência geral de ESBL chegou a 22 por cento, somando-se todos os isolados produtores. O HUM, mesmo sendo considerado um hospital de pequeno porte, apresenta níveis altos de resistência a antimicrobianos, semelhantes àqueles observados em hospitais maiores, demonstrando a necessidade de cuidadosa vigilância epidemiológica.

Relato de caso de febre tifóide no Município de Maringá, Estado do Paraná / Report on typhoid fever case in Maringá, State of Paraná

A febre tifóide é doença bacteriana aguda causada por Salmonella enterica sorotipo typhi, que é adquirida pela ingestão de água ou alimento contaminado. O objetivo do presente trabalho é descrever um caso de febre tifóide ocorrido em Maringá, após três anos sem notificação da doença no Estado do Paraná.

Typhoid fever is an acute bacterial disease caused by Salmonella enterica serotype typhi, which is acquired by consumption of contaminated food or water. This paper had the aim of describing a case of typhoid fever that occurred in Maringá, State of Paraná, after three years without any notifications of the disease.

Mycoplasma hominis e Ureaplasma sp. em amostras do trato genitourinário e sua relação com sintomas de infecção genital / Mycoplasma hominis and Ureaplasma sp. in genitourinary specimens and their association with symptoms of genital infection

Mycoplasma hominis e Ureaplasma sp. são espécies de micoplasmas comensais pertencentes à microflora do trato geniturinário humano. No entanto, existem evidências do papel etiológico destas bactérias em diversas infecções. O objetivo deste estudo foi avaliar a ocorrência de M. hominis e Ureaplasma sp. em pacientes atendidos no Laboratório de Ensino e Pesquisa em Análises Clínicasde Maringá – PR e sua relação com sintomas de infecção genital. Amostras genitais de 1553 pacientes foram pesquisadas para micoplasmas e outros agentes comumente envolvidos em doenças sexualmente transmissíveis (DST). Positividade para micoplasma, em concentração clinicamente significativa, foi observada em 47,1 dos pacientes. O isolamento exclusivo de M. hominis ou Ureaplasma sp. foi observado em 36 pacientes (2,32) e 536 (34,51), respectivamente. Em 159 pacientes (10,24) foram isoladas ambas as espécies. Da totalidade de mulheres estudadas, 1371 (94,5) relataram sintoma de infecção genital e destas 678 (49,5) apresentaramcultura positiva para micoplasma. A alta positividade de M. hominis e Ureaplasma sp. e a associação com sintomas de infecção genital, observada neste estudo, pode contribuir para evidenciar a importância destes microrganismos como agentes de infecção genital.

Acompanhamento farmacoterapêutico de crianças sob prescrição de antimicrobianos: um estudo em farmácia comunitária / Pharmacotherapeutic follow-up of children under antimicrobial prescription: a community pharmacy study

Atualmente, os antibióticos estão entre os medicamentos prescritos com maior freqüência, tanto em ambulatórios quanto em hospitais. As infecções respiratórias agudas são importante causa para atenção médica em atendimento primário à saúde nos países em desenvolvimento, para as quais há necessidade de se garantir uma apropriada adesão, não somente para evitar falhas terapêuticas, mas também para diminuir os riscos de complicações. O estudo teve como objetivo identificar variáveis sociais e fármaco-epidemiológicas, por meio do acompanhamento de cuidadores de crianças responsáveis pela administração de antimicrobianos prescritos. O estudo foi prospectivo e descritivo, desenvolvido em uma farmácia comunitária da cidade de Ibiporã-PR, de maio a julho de 2003. A amostra foi de 85 crianças de zero a 15 anos de idade.A maioria dos cuidadores possuía no mínimo oito anos de escolaridade e a administração do antibiótico era realizada pela mãe/pai. Observou-se que para a amoxicilina, administrada entre o 1º e 3º dia, a categoria "nenhuma queixa" foi referidapor 83% dos cuidadores, enquanto para a cefalexina, por 70%; entre o 4º e 7º dia, para a amoxicilina foi de 81% e de 86% para a cefalexina. A ocorrência de reações adversas a medicamentos (RAM) foi de 30% para a cefalexina e de 19% para a amoxicilina. No 5o dia 44% das crianças haviam interrompido o tratamento com amoxicilina e 14% com cefalexina. Estes achados reforçam a necessidade de acompanhamento farmacoterapêutico sistematizado

Currently, antibiotics are among the most often prescribed medication, as in ambulatories as in hospitals. The acute respiratory infections are an important issue for the medical care regarding fi rst health care in developing countries where there is a need of guaranteeing appropriate adherence, not only to avoid therapeutic fl aw, but also to decrease the risk of complications. The goal of the research was to identify social and pharmacoepidemiological variables, and to accompany children's caretakers responsible for the administration of antimicrobial prescription. The study was prospective and descriptive, and developed in a community pharmacy in Ibiporã (Paraná State, Brazil) from May to July 2003. The sample consisted of 85 children (0 to 15 years old). Most caretakers had at least eight years of schooling and the antibiotics were administered by the mother/father. For Amoxicilin, administered between the 1st and 3rd, day 83% of the caretakers mentioned the "no complaints" category, as 70% for Cephalexin.When administered between the 4th and 7th day, the results were 81 and 86%, respectively. The occurrence of adverse drug reactions (ADR) was 30% for cephalexin (1st and 3rd day) and 19% for Amoxicilin (4th and 7th day). By the fi fth day, 44% of the children had interrupted the Amoxicilin treatment and 14% the Cephalexin therapy. These findings reinforce the need for systematized pharmacotherapeutic follow-up

Pharmacological studies of Cordia salicifolia Cham in normal and diabetic rats

O efeito anti-obesidade, anorexigeno, diurético e anti-hiperlipêmico de folhas secas pulverizadas de Cordia salicifolia Cham (CS) foi investigado. CS foi dissolvida em água e administrada (20 mg.Kg-1.dia-1) por gavagem durante 13 dias. A dose utilizada correspondente à empregada no Brasil. Contrariamente ao uso popular, o efeito anti-obesidade, anorexigeno e diurético não foi observado. Porém, observamos efeito hipolipidêmico em ratos normais e portadores de diabetes induzidos por aloxana.

Absence of antidiabetic and hypolipidemic effect of Gymnema sylvestre in non-diabetic and alloxan-diabetic rats

Neste estudo foi investigado o potencial antidiabético e hipolipemiante da Gymnema sylvestre (GS). O efeito agudo da GS administrada via oral sobre a glicemia e lipidemia foi avaliado em ratos Wistar sob as seguintes condições: a) após refeição balanceada; b) após ingestão de 1000 mg de amylose ou 1000 mg/kg de glicose; c) após ingestão de uma mistura de 12 mL/kg de óleo de soja + 1% de colesterol (OSC). Foi avaliado também o efeito do tratamento com GS durante duas (sub-agudo) ou quatro semanas (crônico) sobre o peso corporal, ingestão de água e alimento, glicemia e lipidemia em ratos não diabéticos e diabéticos induzidos com aloxana. A dose de GS utilizada na maioria dos experimentos (30 mg/kg) corresponde a dose empregada para tratar diabetes no Brasil. A administração aguda de GS não influenciou a elevação da glicemia durante a refeição balanceada, administração de amilose ou glicose; mas intensificou a elevação da concentração sérica de lipídeos após a administração de OSC. Além disso, o tratamento sub-agudo e crônico com GS em ratos não diabéticos e diabéticos induzidos com aloxana não alterou: a) o ganho de peso corporal; b) a ingestão de água e alimento; c) os níveis de glicose e lipídios. Assim sendo, a Gymnema sylvestre pelo menos na forma comercializada no Brasil (folhas secas em pó) requer mais ensaios experimentais e clínicos antes de ser recomendada para tratamento do diabetes e hiperlipidemia.

Estudo do perfil de susceptibilidade antimicrobiana e avaliação molecular de amostras de Enterococcus spp isoladas de pacientes hospitalizados / Study of drugs susceptibility and molecular samples evaluation of Enterococcus spp isolated in hospitalized patients

Neste estudo foi investigado o perfil de susceptibilidade antimicrobiana e avaliação molecular de amostras de Enterococcus spp, isoladas de urina, secreção purulenta, sangue e ferida cirúrgica de pacientes do Hospital Universitário de Maringá, estado do Paraná, no período entre 1998 e 2000. As amostras foram avaliadas quanto à susceptibilidade a drogas, tipificadas através de JB1-PCR e investigadas quanto à presença de gene Van A e Van B. Das 58 amostras, 98,3 por cento foram classificadas como Enterococcus faecalis. Todas as amostras foram sensíveis à nitrofurantoína e vancomicina, 96,6 por cento sensíveis à ampicilina, 32,8 por cento foram resistentes à estreptomicina, 10,3 por cento resistentes à gentamicina e ciprofloxacina e 6,9 por cento resistentes a norfloxacina. Por meio do teste de CIM, para vancomicina, observou-se crescimento bacteriano até a concentração de 4μg/ml. Não foi detectada a presença de gene Van A e Van B. A amplificação por JB1-PCR proporcionou padrões de bandas com até 7 fragmentos sendo as amostras divididas em 6 grupos distintos. Não houve prevalência de algum padrão de bandeamento que denotasse a presença de um clone e não foi possível estabelecer correlação entre genótipo com resistência antimicrobiana