Abstract INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is a common pathogen causing healthcare-associated infections. Owing to the restricted use of beta-lactams in MRSA infections, non-beta-lactam antimicrobials are required for treatment. However, MRSA can develop resistance mechanisms to non-beta-lactam antimicrobials, which reduces viable treatment options. Here, we evaluated the antimicrobial susceptibility and resistance genes of MRSA isolated from hospitalized patients in South Brazil. METHODS: The antimicrobial susceptibilities of hospital MRSA (217) isolates were determined by disk diffusion or microdilution methods. Additionally, the presence of 14 resistance genes and SCCmec typing was performed by PCR. RESULTS: Among the antimicrobials tested, we observed high erythromycin (74.2%), ciprofloxacin (64.5%), and clindamycin (46.1%) resistance rates and complete susceptibility to linezolid and vancomycin. Seventeen different patterns of MRSA antimicrobial resistance were observed, of which 42.9% represented multidrug resistance. Among erythromycin-resistant MRSA, 53.4%, 45.3%, 37.9%, 13.0%, and 6.8% carried ermA, msrA, msrB, ermC, and ermB genes, respectively. Among clindamycin-resistant MRSA, 83%, 17%, 10%, 4%, and 2% carried ermA, ermC, ermB, linA, and linB genes, respectively. Among gentamicin-resistant MRSA, 96.8%, 83.9%, and 9.7% carried aac(6')/aph(2''), aph(3’)-IIIa, and ant(4’)-Ia genes, respectively. Among tetracycline-resistant MRSA, 6.5% and 93.5% carried tetK and tetM genes, respectively. Lastly, among trimethoprim/sulfamethoxazole-resistant MRSA, 13.3% and 100% carried dfrA and dfrG genes, respectively. The SCCmec type IV isolates were detected more frequently, whereas the SCCmec type III isolates exhibited higher multidrug resistance. CONCLUSIONS: The study data provides information regarding the MRSA resistance profile in South Brazil that is associated with the clinical conditions of patients and can contribute to clinical decision-making.

In this study, we aimed to evaluate the performance of different phenotypic tests to detect carbapenem-resistant Enterobacteriaceae. Three different phenotypic methods were evaluated: (1) combined-disk test of meropenem plus phenylboronic acid or EDTA reading after 24 h and 48 h; (2) selective/chromogenic read after 24 h and after 48 h; and (3) overnight selective enrichment broth containing 10 µg ertapenem disk followed by culture on MacConkey agar. A positive result in at least one of the methods was submitted to PCR for blaNDM-1, blaOXA-48, blaKPC, blaSPM-1, blaIMP, and blaGES detection. Carbapenem-resistant Enterobacteriaceae was detected in 31 (30.4%) of 102 rectal swabs evaluated. All isolates showed to be KPC-2-producing organisms. Results showed excellent agreement among the evaluated tests (positive and negative) (kappa = 0.88). It is important to state that combined-disk test with phenylboronic acid is not suitable for bacterial identification/isolation. Conversely, selective/chromogenic agar after 48h of incubation showed to be a useful tool, with the advantage of presumptive bacterial identification.

Helicobacter pylori infection is one of the most common infections worldwide and is associated with gastric diseases. Virulence factors such as VacA and CagA have been shown to increase the risk of these diseases. Studies have suggested a causal role of CagA EPIYA-C in gastric carcinogenesis and this factor has been shown to be geographically diverse. We investigated the number of CagA EPIYA motifs and the vacA i genotypes in H. pylori strains from asymptomatic children. We included samples from 40 infected children (18 females and 22 males), extracted DNA directly from the gastric mucus/juice (obtained using the string procedure) and analysed the DNA using polymerase chain reaction and DNA sequencing. The vacA i1 genotype was present in 30 (75%) samples, the i2 allele was present in nine (22.5%) samples and both alleles were present in one (2.5%) sample. The cagA-positive samples showed distinct patterns in the 3’ variable region of cagA and 18 of the 30 (60%) strains contained 1 EPIYA-C motif, whereas 12 (40%) strains contained two EPIYA-C motifs. We confirmed that the studied population was colonised early by the most virulent H. pylori strains, as demonstrated by the high frequency of the vacA i1 allele and the high number of EPIYA-C motifs. Therefore, asymptomatic children from an urban community in Fortaleza in northeastern Brazil are frequently colonised with the most virulent H. pylori strains.

Double disks synergy test (DDST) and combined disks test (CD) were evaluated to predict the presence of metallo-β-lactamase in 70 Pseudomonas aeruginosa isolates recovered from cystic fibrosis and non-cystic fibrosis patients. DDST CAZ-EDTA 1 cm and CD IMP-EDTA tests showed the best accuracy (94.3%). Furthermore, for other combinations, accuracy unsatisfactory was obtained.

INTRODUÇÃO: Vigilância com base em detecção laboratorial é um componente importante no controle de enterococos resistentes a vancomicina (ERV). MÉTODOS: Avaliamos procedimento da reação em cadeia da polimerase real time (PCR-RT) (genes vanA-vanB) para detecção de ERV em 115 swabs de pacientes incluídos em um programa de vigilância. RESULTADOS: A sensibilidade do RT-PCR foi semelhante a da cultura primária (75% e 79,5%, respectivamente) quando comparada com a cultura em caldo enriquecido, enquanto a especificidade foi de 83,1%. CONCLUSÕES: O RT-PCR fornece resultados no mesmo dia, contudo mostra baixa sensibilidade para a detecção de VRE.

INTRODUCTION: Laboratory-based surveillance is an important component in the control of vancomycin resistant enterococci (VRE). METHODS: The study aimed to evaluate real-time polymerase chain reaction (RT-PCR) (genes vanA-vanB) for VRE detection on 115 swabs from patients included in a surveillance program. RESULTS: Sensitivity of RT-PCR was similar to primary culture (75% and 79.5%, respectively) when compared to broth enriched culture, whereas specificity was 83.1%. CONCLUSIONS: RT-PCR provides same day results, however it showed low sensitivity for VRE detection.

Staphylococcus aureus resistente à meticilina (MRSA), outrora isolado especificamente em ambientes hospitalares, vem sendo identificado como causador de infecções cutâneas em pacientes da comunidade. Neste artigo, é relatado um caso do sul do Brasil com furunculose por CA-MRSA. O microrganismo isolado foi submetido a exames de PCR para o gene mecA e para o gene que codifica a leucocidina de Panton-Valentine. Esses exames permitiram a identificação genotípica do CA-MRSA.

Methicillin-resistant Staphylococcus aureus (MRSA), particularly isolated at hospital setting, has been identified in cutaneous infections of community patients. This paper reports a case of furunculosis from the southern Brazil. Dermatologists must be attentive to this emergent etiological diagnosis. The isolated microorganism was subjected to PCR for gene mecA and to PCR for the gene that encodes the leukocidin of Panton-Valentine. These exams enabled genotypic identification of CA-MRSA.

Enterococci are part of the endogenous flora of human beings, are naturally resistant to several classes of antimicrobials, and are able to acquire resistance with relative ease. Currently the vancomycin-resistant enterococci are spread all over the world and treatment options for infections caused by it are often extremely limited. We assessed 193 vancomycin-resistant Enterococcus faecalis isolates collected from four different hospitals in Porto Alegre for their susceptibility to fosfomycin using the E-test and agar diffusion. Fosfomycin proved to be active in vitro against the great majority of isolates, indicating that it is a valid option in the treatment of these infections.

Foram estudadas 455 amostras de enterococos isolados de pacientes moradores da cidade de Porto Alegre, Rio Grande do Sul, Brasil, durante o período de julho 1996 a junho 1997 e foram identificados ao nível de espécies por testes fisiológicos convencionais e analisados sua susceptibilidade aos agentes antimicrobianos. A diversidade genética foi avaliada por eletroforese de campo pulsado ("pulsed-field gel electrophoresis", PFGE) com a enzima de restrição SmaI. A espécie mais freqüente encontrada foi o Enterococcus faecalis (92,8%). As outras espécies identificadas foram: E. faecium (2,9%), E. gallinarum (1,5%), E. avium (1,1%), E. hirae (0,7%), E. casseliflavus (0,4%), E. durans (0,4%) and E. raffinosus (0,2%). A prevalência de amostras com níveis elevados de resistência (HLR) aos aminoglicosídeos foram de 37,8%. HLR para gentamicina foi encontrada em 24,8% das amostras. Nenhuma amostra com resistência adquirida à vancomicina foi isolada. A análise através de PFGE revelou a predominância do grupo clonal A, constituído por amostras isoladas de diferentes materiais clínicos obtidos de pacientes internados em três hospitais. Esses resultados sugerem a disseminação intra e inter-hospital de um clone predominante, composto por amostras de E. faecalis apresentando níveis elevados de resistência a gentamicina, nos hospitais incluídos neste estudo.

In the present study, a total of 455 enterococcal isolates, recovered from patients living in the city of Porto Alegre, State of Rio Grande do Sul, Brazil, during the period from July 1996 to June 1997, were identified to the species level by conventional biochemical and microbiological tests, and assayed for their susceptibilities to antimicrobial agents. The genetic diversity of antimicrobial resistant strains was evaluated by pulsed-field gel electrophoresis (PFGE) analysis of SmaI restricted chromosomal DNA. The most frequent species was Enterococcus faecalis (92.8%). Other species identified were: E. faecium (2.9%), E. gallinarum (1.5%), E. avium (1.1%), E. hirae (0.7%), E. casseliflavus (0.4%), E. durans (0.4%) and E. raffinosus (0.2%). The overall prevalence of isolates with high-level resistance (HLR) to aminoglycosides was 37.8%. HLR to gentamicin was found in 24.8%. No strains with acquired resistance to vancomycin were found. PFGE analysis showed the predominance of clonal group A, comprising strains isolated from different clinical specimens obtained from patients in three hospitals. These results suggest intra and inter-hospital dissemination of one predominant clonal group of E. faecalis isolates with HLR to gentamicin in the hospitals included in this study.

OBJETIVO: O objetivo deste estudo foi determinar a prevalência do S. pneumoniae resistente aos macrolídeos e identificar suas características fenotípicas e genotípicas. MÉTODOS: Amostras de S. pneumoniae isoladas entre maio de 2002 e agosto de 2004, em Porto Alegre (RS), a partir de materiais clínicos coletados de diferentes sítios anatômicos foram analisadas. Para o teste de difusão em ágar foram utilizados discos de eritromicina, claritromicina, azitromicina e clindamicina. As concentrações inibitórias mínimas de eritromicina foram determinadas nos isolados resistentes aos macrolídeos pelo método de diluição em ágar. Os fenótipos dos isolados resistentes aos macrolídeos foram investigados pelo teste de difusão em ágar e a genotipagem pela reação em cadeia da polimerase. RESULTADOS: Foram avaliados 229 isolados de pneumococos, e 12 mostraram-se resistentes aos macrolídeos (5,2%). Entre estes, 9 apresentaram o fenótipo MLSB (75%) e 3 o fenótipo M (25%). A reação em cadeia da polimerase indicou que 8 isolados com o fenótipo MLSB portavam apenas o gene ermB, enquanto que o gene mefE estava presente em todos os 3 isolados com o fenótipo M. Um isolado com o fenótipo MLSB apresentou ambos os genes. CONCLUSÃO: A resistência aos macrolídeos do S. pneumoniae em Porto Alegre permanece baixa, sendo devida principalmente à presença do gene ermB, com expressão do fenótipo MLSB.

OBJECTIVE: The aim of this study was to determine the prevalence of macrolide-resistant S. pneumoniae and to identify its phenotypic and genotypic characteristics. METHODS: Strains of S. pneumoniae isolated in the city of Porto Alegre between May 2002 and August 2004 from samples collected from different anatomical sites were analyzed. For the agar diffusion test, disks of erythromycin, clarithromycin, azithromycin and clindamycin were used. The minimum inhibitory concentrations of erythromycin were determined for macrolide-resistant isolates by the agar dilution method. Macrolide-resistant isolates were phenotyped by agar diffusion test and genotyped by polymerase chain reaction. RESULTS: A total of 229 pneumococcal strains were evaluated, 12 (5.2%) of which were macrolide-resistant. Among the 12 resistant strains, 9 (75%) presented the MLSB phenotype, and 3 (25%) presented the M phenotype. Polymerase chain reaction testing indicated that 8 MLSB phenotype isolates harbored the ermB gene only, whereas the mefE gene was present in all 3 M phenotype isolates. One MLSB phenotype isolate presented both genes. CONCLUSION: In Porto Alegre, the S. pneumoniae resistance to macrolides is still low since such resistance is due primarily to the presence of the ermB gene expressing the MLSB phenotype.

A resistência a várias classes de agentes antimicrobianos é uma característica marcante dos enterococos observada em diferentes regiões geográficas. Informações sobre amostras isoladas na região sul do Brasil ainda são limitadas. No presente estudo, 455 enterococos isolados consecutivamente de pacientes moradores na cidade de Porto Alegre, Brasil, foram identificados ao nível de espécie e testados em relação a sua sensibilidade aos antimicrobianos através de testes de difusão em agar. As espécies mais freqüentes foram E. faecalis (92,8%), seguidas por E. faecium (2,9%), E. gallinarum (1,5%), E. avium (1,1%), E. hirae (0,7%), E. casseliflavus (0,4%), E. durans (0,4%) e E. raffinosus (0,2%). Os testes de sensibilidade revelaram que 62,0% das amostras foram resistentes à tetraciclina, 42,6% à eritromicina, 24,8% ao cloranfenicol, 22,6% à ciprofloxacina, 22,0% à norfloxacina, 3,5% à ampicilina e 3,5% á nitrofurantoína. Resistência a níveis elevados de aminoglicosídeos foi encontrada em 37,8% das amostras, com 23,5% sendo resistente à gentamicina, 14,3% à estreptomicina e 2,8% a ambos gentamicina e estreptomicina. Nenhuma amostra com resistência adquirida à vancomicina ou produtora de b-lactamase foi encontrada. Os resultados indicam um significante percentual de amostras resistentes a diferentes antimicrobianos, apontando a necessidade de estratégias de controle para evitar a disseminação de cepas resistentes e de vigilância contínua para a detecção de características emergentes de resistência.

Resistance to several classes of antimicrobial agents is a remarkable characteristic of enterococcal strains increasingly reported worldwide. Information about strains isolated in the southern region of Brazil is still limited. In this study, a total of 455 consecutive enterococcal isolates recovered from patients living in Porto Alegre, Brazil, were identified at species level and evaluated for their antimicrobial susceptibilities by agar diffusion testing. The most frequent species was E. faecalis (92.8%), followed by E. faecium (2.9%), E. gallinarum (1.5%), E. avium (1.1%), E. hirae (0.7%), E. casseliflavus (0.4%), E. durans (0.4%), and E. raffinosus (0.2%). According to the results of disk tests 62.0% of the strains were resistant to tetracycline, 42.6% to erythromycin, 24.8% to chloramphenicol, 22.6% to ciprofloxacin, 22.0% to norfloxacin, 3.5% to ampicillin, 3.5% to nitrofurantoin. High level resistance to aminoglycosides was found in 37.8% of the isolates, with 23.5% being resistant to gentamicin, 14.3% to streptomycin, and 2.8% to both gentamicin and streptomycin. No vancomycin resistant or b-lactamase producing isolates were found. The results indicate that a significant percentage of isolates are resistant to different antimicrobials, pointing out the need for control strategies to avoid dissemination of resistant isolates and for continuous surveillance for the detection of emerging resistance traits.

O uso de métodos automatizados tem freqüentemente levado a falhas na identificação do gênero Enterococcus em laboratórios de microbiologia clínica. Neste estudo foi avaliada a utilização de um sistema automatizado Vitek (bioMérieux) em dois laboratórios de microbiologia clínica para identificação de diferentes espécies de enterococos. Os resultados foram comparados com os testes fisiológicos convencionais. As amostras (80) foram inoculadas em testes bioquímicos convencionais e no cartão Vitek GPI. No geral, a concordância entre os dois métodos foi de 83,7% (67/80). Entre as amostras de E. faecalis, o sistema Vitek identificou corretamente 35/40 (87,5%) e entre os E. faecium, a concordância foi 12/14 (85,7%). Em 20/26 amostras (76,9%) pertencentes a espécies não-E. faecalis e não-E. faecium, o sistema chegou à identificação correta. Os resultados do presente estudo mostram que o sistema Vitek necessita de melhorias para a identificação de enterococos, especialmente diante de espécies menos freqüentes.

Automated systems may present problems in the identification of members of the genus Enterococcus in clinical laboratories. Having conventional physiological tests as the reference method, we evaluated the use of an automated system (VITEK - bioMérieux) in the identification of 80 isolates belonging to different species of Enterococcus. The general agreement between results obtained by the conventional method and by the Vitek GPI card was 83.7% (67/80). Among isolates of E. faecalis and E faecium we observed that the automated system correctly identified 35/40 (87.5%) and 12/14 (85.7%) of the strains, respectively. Among isolates belonging to species which are neither E. faecalis, nor E. faecium, it was observed an agreement of 20/26 (76.9%). Results point to the need of improvement in the automated systems to identify enterococci. Special consideration must be taken regarding less frequently isolated species.

O Rhodococcus equi, principal agente da rodococose, é um cocobacilo pleomórfico, gram-positivo e aeróbio, que infecta humanos por via inalatória ou transcutânea e se manifesta clinicamente como abscesso pulmonar. Relatam-se os dois primeiros casos brasileiros da doença. Ambos os pacientes eram imunocomprometidos e apresentavam quadro infeccioso pulmonar. O primeiro tinha AIDS e apresentava pneumonia cavitada em lobo superior esquerdo, que teve evolução fatal. O segundo tinha doença de Goodpasture, insuficiência renal crônica e fazia uso de corticosteróides. Apresentava uma lesão pulmonar escavada no lobo superior direito, que foi tratada com sulfametoxazol-trimetoprim, com resolução do processo.

Rhodococcus equi, the principal agent of rhodococcosis, is a pleomorphic, gram-positive, aerobic coccus bacillus that infects humans by inhalation or through a transcutaneous route. It is clinically manifested as a pulmonary abscess. The first two Brazilian cases of rhodococcosis are reported on. Both patients were immunocompromised and showed pulmonary infection. The first patient had AIDS and cavitating pneumonia in the left upper lobe, that was fatal. The second case presented Goodpasture syndrome and was under chronic corticotherapy. He displayed a cavitating nodular lesion in the right upper lobe, that was successfully treated with sulfametoxazol-trimethoprim.

Os autores relatam três casos de neurocriptococose em crianças na primeira década de vida, no Estado do Rio Grande do Sul. O diagnóstico foi estabelecido pelo encontro do fungo Cryptococcus neoformans no líquido céfalo-raquidlano por exame direto com tinta da China e cultura em meio de Sabouraud. As formas de instalação da síndrome neurológica foram aguda, subaguda e crônica. Síndrome de hipertensão intracraniana e hidrocefalia foram complicações observadas durante a evolução da doença, sendo necessária a instalação de derivação ventrículo-peritonial em um caso. Desnutrição foi o processo mórbido associado nos três casos e infecção pelo bacilo da tuberculose em um caso. Terapia específica foi administrada aos três pacientes. Um paciente evoluiu para cura e dois evoluíram para óbito.

The authors report three cases of central nervous system cryptococcosis in children in the first decade of life, from Rio Grande do Sul State, Brazil. Diagnosis was supported by demonstration of Cryptoccoccus neformans with India-ink preparations of the spinal fluid and Sabouraud's media culture. Clinical presentation included acute, subacute and chronic forms. Increased intracranial pressure and hydrocephalus were the complications during the course of the disease, and a ventriculoperitoneal shunt was used in one case. Undernutrition was associated with all cases and tubercle bacillus infection with one. Specific therapy was administered to the three patients. One patient was cured and the other two died.