Abstract Identification of nonfermenting Gram-negative bacteria (NFGNB) of cystic fibrosis patients is hard and misidentification could affect clinical outcome. This study aimed to propose a scheme using polymerase chain reaction to identify NFGNB. This scheme leads to reliable identification within 3 days in an economically viable manner when compared to other methods.

RESUMO: Salmonella Gallinarum (S. Gallinarum) e Salmonella Pullorum (S. Pullorum) são patógenos hospedeiro-específico de aves, agentes do tifo aviário e pulorose, respectivamente. Estes biovares causam infecções septicêmicas, resultando em alta mortalidade. Surtos são frequentemente relatados em diversos países, causando prejuízos devido à eliminação de lotes infectados e tratamentos. Agentes antimicrobianos são utilizados frequentemente em granjas avícolas para prevenir ou tratar infecções gastrointestinais. No presente trabalho, foi avaliada a susceptibilidade antimicrobiana de 50 isolados de S. Gallinarum e S. Pullorum, obtidos durante surtos que ocorreram entre 1987 a 1991 e entre 2006 a 2013. A comparação dos perfis de sensibilidade mostrou que todas as amostras são sensíveis aos β-lactâmicos. Todos os isolados de 1987-1991 foram sensíveis a todos os antibióticos testados, exceto NAL e CIP (78%). O perfil de susceptibilidade de S. Gallinarum (surtos de 2006 a 2013) foi NAL (58%), CIP (63%), ENR (67%), TET (92%), FFC (96%) e SXT (96%). Isolados de S. Pullorum (surtos de 2006 a 2013) apresentaram as seguintes taxas de sensibilidade: NAL (65%), CIP (71%), ENR (94%) e TET (94%). Todas as amostras foram sensíveis ao β-lactâmicos testados, no entanto, a resistência às quinolonas e fluoroquinolonas aumentou ao longo do tempo. Além disso, baixos níveis de resistência a outros antibióticos foram encontrados em isolados recentes, tais como as tetraciclinas.

ABSTRACT: Salmonella Gallinarum (S. Gallinarum) and Salmonella Pullorum (S. Pullorum) are poultry host-specific, agents of fowl typhoid and pullorum disease, respectively. These biovars cause septicemic infections, resulting in high mortality. Outbreaks are frequently reported worldwide, causing losses due to the elimination of infected flocks and treatments. The use of antimicrobial agents is frequent in poultry farms to prevent or treat gastrointestinal infections. In the present research it was evaluated the antimicrobial susceptibility of 50 S. Gallinarum and S. Pullorum isolates, from outbreaks that occurred between 1987 to 1991 and 2006 to 2013. The comparison of the susceptibility profiles showed that all isolates were susceptible to β-lactams. All isolates from 1987-1991 were susceptible to all antibiotics tested except NAL and CIP (78%). The susceptibility profile of S. Gallinarum (2006 - 2013 period) was the following NAL (58%), CIP (63%), ENR (67%), TET (92%), FFC (96%) and SXT (96%). S. Pullorum isolates (2006 - 2013 period) showed the following susceptibility rates to NAL (65%), CIP (71%), ENR (94%) and TET (94%). All isolates were susceptible to β-lactams tested, however, resistance to quinolones and fluoroquinolones increased over time. Furthermore, low levels of resistance to other antibiotics were found in recent isolates, such as tetracyclines.

INTRODUCTION:Polymyxins are antimicrobial agents capable of controlling carbapenemase-producing Klebsiella pneumoniae infection.METHODS: We report a cluster of four patients colonized or infected by polymyxin-resistant and Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae.RESULTS: Three patients were hospitalized in adjacent wards, and two were admitted to the intensive care unit. The index case maintained prolonged intestinal colonization by KPC-producing K. pneumoniae. Three patients received polymyxin B before the isolation of polymyxin-resistant K. pneumoniae.CONCLUSIONS: Colonization by KPC-producing K. pneumoniae and previous use of polymyxin B may be causally related to the development of polymyxin-resistant microorganisms.

This study evaluated the relationship between previous colonization of the oropharynx and development of ventilator-associated pneumonia through the classification of genomic fingerprint pattern by pulsed-field gel electrophoresis of both oxacillin-resistant and oxacillin-susceptible Staphylococcus aureus isolates obtained from hospitalized patients in an intensive care unit.

Background: The carrier state of methicillin-resistant Staphylococcus aureus is pointed as an infection predictor and a factor for environmental and person-to-person dissemination, including health service workers. These, when colonized are commonly associated to outbreaks. Objective: Analyze the prevalence of Staphylococcus aureus in saliva of workers at a university hospital. Methodology: Epidemiologic longitudinal study carried out in Curitiba, Paraná, Brazil, with 486 workers between April 2006 and June 2008. Three saliva samples were collected and a data collection instrument was applied. Staphylococcus aureus were isolated from the clinical specimen and characterized by phenotypes. The data from the instrument and the laboratory results were organized and processed with EPI-Info software and analyzed via descriptive statistics. Results: Among the healthcare workers studied, 60.9% were colonized by Staphylococcus aureus in saliva; of those, 67.9% were transitory carriers and 32.1% were persistent carriers; the prevalence of meticillin-resistant Staphylococcus aureus (MRSA) among the isolated cases was 15.7%. The average prevalence of MRSA was 12.7% and higher among nurses’ aides (21.4%) and cleaning aides (20.6%) and lower among nurses (4.5%) and doctors (5.9%). Conclusions: Healthcare workers presented high prevalence of Staphylococcus aureus in saliva, indicating the mouth as an important body site to investigate colonization by MRSA and a potential source to its dissemination.

Introdução: O estado de portador de Staphylococcus aureus resistente à meticilina é apontado como preditor de infecção e fator para a disseminação ambiental e de pessoa a pessoa, incluindo trabalhadores de serviço de saúde. Estes quando colonizados são freqüentemente associados a surtos. Objetivo: Analisar a prevalência de Staphylococcus aureus na saliva de trabalhadores de hospital universitário. Metodologia: Estudo epidemiológico longitudinal realizado em Curitiba, Paraná, Brasil, com 486 trabalhadores no período de abril de 2006 a junho de 2008 compreendeu a coleta de três amostras de saliva e aplicação de instrumento de coleta de dados. Staphylococcus aureus foram isolados dos espécimes clínicos e caracterizados fenotipicamente. Os dados foram organizados e processados no Programa EPI-Info e analisados por meio de estatística descritiva. Resultados:Entre os trabalhadores investigados, 60,9% estavam colonizados por Staphylococcus aureus na saliva, sendo 67.9% carreadores transitórios e 32.1% carreadores persistentes; a prevalência de Staphylococcus aureus resistente à meticilina (MRSA) entre os isolados foi de 15.7%. A prevalência média de MRSA foi de 12.7% sendo maior entre técnicos em enfermagem (21.4%) e auxiliares de limpeza (20.6%) e menor entre enfermeiros (4.5%) e médicos (5.9%). Conclusões: Os trabalhadores apresentaram alta prevalência de Staphylococcus aureus na saliva, indicando a boca como importante sítio corporal para a investigação da colonização por MRSA e potencial fonte para sua disseminação.

In this study we report the first isolation of VanA-type vancomycin-resistant Enterococcus faecalis strains from two different patients hospitalized in the same intensive care unit at the hospital of Universidade Federal do Triângulo Mineiro (UFTM), Uberaba, Minas Gerais, Brazil.

Staphylococcus aureus and coagulase-negative staphylococci are the main cause of sepsis in Neonatal Intensive Care Unit (NICU). Central venous catheters (CVCs) are an important part of critical neonates' treatment and are associated with sepsis. The aim of this study was to investigate two outbreaks caused by Staphylococcus aureus and Staphylococcus epidermidis associated with CVC inserted by phlebotomy in critical neonates. The surveillance was performed from January 2001 to December 2005 at the Brazilian NICU. The genotypic analysis of oxacillin susceptible S. aureus (OSSA) and oxacillin resistant S. epidermidis (ORSE) was performed based on pulsed-field gel electrophoresis (PFGE). Staphylococcus was the most frequent pathogen (65.8%) with highest incidence of CoNS (59.9%) followed by S. aureus (40.1%). During the five years of surveillance, there were two outbreaks detected, occurred in January-February/02 and August/02 and confirmed by PFGE analysis. The predisposing factors for infection corresponding to both outbreaks were: age <7 days, hospitalization > 7 days, and use of polyethylene CVC through dissection of vein (phlebotomy). This is the first relate of staphylococcal outbreaks associated with CVC inserted by phlebotomy in NICU. PFGE showed polyclonal spread of OSSA during both epidemic and endemic period, and two monoclonal outbreaks of ORSE in the same epidemic period of OSSA.

A identificação das espécies de 52 Enterococcus spp. isolados de amostras de alimentos foi realizada empregando-se duas metodologias: sistema API 20 STREP e PCR multiplex. Os resultados obtidos revelaram que 78,9% dos isolados apresentaram resultados diferentes nos dois testes utilizados. Apenas seis E. faecalis e cinco E. faecium apresentaram resultados concordantes pelos dois métodos. PCR multiplex permitiu a identificação completa de um número significantemente maior de enterococos do que o sistema API 20 STREP.

We evaluated the suitability of API 20 STREP and multiplex PCR to speciate 52 Enterococcus spp. obtained from Brazilian foods. A high percentage of isolates (78.9%) presented discrepant results between evaluated tests. Similar results were obtained for six E. faecalis and five E. faecium. The PCR multiplex was more effective than API 20 STREP for complete identification of the isolates.

We report the first case of vancomycin-resistant Enterococcus faecalis with VanA phenotype and vanA genotype isolated from a surgical patient in a general Hospital in Uberlândia, state of Minas Gerais, Brazil.

The isolation of vancomycin resistant enterococci (VRE) in Brazil has rapidly increased, following the world wide tendency. We report in the present study the first isolation of vancomycin resistant Enterococcus faecalis (VRE) in the Northeast of Brazil. The four VRE isolates were characterized for antimicrobial susceptibility, genotypic typing by macro restriction of chromosomal DNA followed by pulsed-field gel electrophoresis and for characterization of the Tn1546-like element and plasmid contents. The isolates showed resistance to multiple antibiotics and a single genotype profile, suggesting the dissemination of a single clone among the patients. Tn1546 associated to genetic elements as plasmids shows the importance of infection control measures to avoid the spreading of glycopetide resistance by conjugative transfer of VanA elements.

No Brasil, enterococos resistente à vancomicina (VRE) têm sido descritos como patógenos hospitalares, desde 1998. Durante um monitoramento de VRE em um hospital, foram detectadas duas cepas de Enterococcus faecalis com genótipo vanA, e sensibilidade à teicoplanina. Este é o primeiro relato do isolamento de enterococo fenótipo VanB e genótipo vanA de amostra clínica no Brasil.

In Brazil, vancomycin-resistant enterococci (VRE) have been reported as nosocomial pathogens since 1998. Recently, in a VRE surveillance in a hospital, we detected two Enterococcus faecalis isolates with vanA genotype and susceptible to teicoplanin. This is the first report of VanB phenotype-vanA genotype enterococci isolated from humans in Brazil.

Staphylococcus aureus e Estafilococos coagulase-negativa (ECN) estão entre os patógenos hospitalares mais importantes em pacientes de unidades de terapia intensiva neonatal, principalmente em infecções da corrente sanguínea. O principal objetivo deste estudo foi determinar a ocorrência de infecções hospitalares por estes microrganismos usando dois sistemas de vigilância (laboratorial e "National Nosocomial Infection Surveillance" - NNIS) e determinar os fatores de risco mais importantes durante o período de dois anos (2001-2002). Dois surtos por ambos S. aureus suscetível a meticilina (1.5%) e ECN resistente à meticilina (1.0%) foram observados, de janeiro a fevereiro/02 e agosto a setembro/02. Taxas de incidência endêmica de 3.77% e 5.16% para S.aureus e ECN foram detectadas respectivamente. Fatores de risco incluíram idade <=7 dias, hospitalização > ou = 7 dias e utilização de cateter vascular central (CVC) de polietileno através de dissecação de veia (flebotomia), mas, nenhum desses fatores independentes foram confirmados pela análise multivariada. Por outro lado, ECN resistente à oxacilina prevaleceu (66.0%) nos episódios epidêmicos. Análise molecular através de gel de eletroforese em campo pulsátil mostrou a natureza policlonal das amostras de S. aureus. Em conclusão, nós identificamos dois surtos de etiologia mista por S. aureus suscetível à meticilina e ECN resistente à meticilina associados à falta de material adequado (cateter vascular central) para neonatos, relacionados a procedimento invasivo. Os dois surtos foram controlados com a substituição de CVC de polietileno pelo cateter central de inserção periférica.

Staphylococcus aureus and coagulase-negative staphylococci (CoNS) are among the most important nosocomial pathogens in patients from neonatal intensive care units, mainly in bloodstream infections. The main objective of this study is to determine the occurrence of nosocomial infections by these microorganisms using two surveillance systems (Laboratorial Surveillance and National Nosocomial Infection Surveillance System) and to determine the most important risk factors during a two-year period (2001-2002). Two outbreaks by both methicillin susceptible S. aureus (MSSA) (1.5%) and methicillin resistant CoNS (MRCoNS) (1.0%) were observed, from January to February/02 and August to September/02. Endemic incidence rates of 3.77% and 5.16% of S. aureus and CoNS, respectively were detected. Risk factors included age <=7 days, hospitalization > or = 7 days and utilization of polietilene central vascular catheter (CVC) through vein dissection (phlebotomy), but none of these independent factors were confirmed by the multivariate analysis. However, oxacillin resistant CoNS prevailed (66.0%) in the epidemic episodes. Molecular analysis by pulsed field gel electrophoresis showed the polyclonal nature of S. aureus isolates. In conclusion, two outbreaks were identified of mixed etiology by MSSA and MRCoNS associated to the lack of an adequate material (central venous catheter) for neonates, related invasive procedure. The outbreaks were controlled with the substitution of polietilene CVC for peripherally inserted central catheter.

We studied an outbreak of two multi-drug resistant clones of Acinetobacter baumannii in the Neonatal Intensive Care Unit of the Uberlândia Federal University Hospital in Minas Gerais state, Brazil, and we analyzed the contribution of cross-transmission in the rise in infection rates. Eleven neonates who developed multi-drug resistant A. baumannii nosocomial infection were matched to 22 neonates who were admitted to the same unit and did not develop an infection during the outbreak period, in order to identify risk factors for infection. Three out of the 11 neonates died. Epidemiological investigation included molecular typing, using pulsed field gel electrophoresis. Prior to the outbreak, from December 2001 to March 2002, no case of infection by this microorganism was diagnosed. Environmental and healthcare worker hand cultures were negative. Nine isolates had similar pulsed field gel electrophoresis patterns and two had another clone. The first clone was brought into the unit by an infected patient who was transferred from another hospital without a history of antibiotic use. The second clone did have its origin clearly defined. Both infected groups led us to conclude that several factors contributed to infection with A. baumannii. These factors were: exposure to antibiotics and invasive devices, birth weight < 1500g, age < 7 days and duration of hospitalization > 7 days. Based on logistic regression, infected neonates were more exposed to carbapenem and mechanical ventilation than the control group. Cross transmission between infants contributed to the rise in the rates of multi-drug resistant A. baumannii infection.

This study reports the results about antimicrobial resistance of Enterococcus spp. isolated from intestinal tract of patients from a university hospital in Brazil. The identification of strains at species level was performed by conventional biochemical tests, API 20 Strep (bioMérieux), and polymerase chain reaction assay. The specie distribution was E. faecium (34%), followed by E. faecalis (33%), E. gallinarum (23.7%), E. casseliflavus (5.2%), E. avium (1%), and E. hirae (1%). Intrinsic resistance to vancomycin characterized by presence of vanC genes was found in E. gallinarum and E. casseliflavus. The high prevalence of VanC phenotype enterococci is very important because these species have been reported as causing a wide variety of infections. Vancomycin-resistant E. faecium or E. faecalis were not found and no one isolate of these species was a beta-lactamase producer. Thirteen clinical isolates of enterococci (13.4%) showed multiresistance patterns, which were defined by resistance to three classes of antibiotics plus resistance to at least one aminoglycoside (gentamicin and/or streptomycin). The resistance to several antimicrobials shown by enterococcal strains obtained in this study is of concern because of the decrease in the therapeutic options for treatment of infections caused by enterococci.

Nesse estudo, para detecção de S. aureus resistente à meticilina (MRSA), a amplificação do gene mecA baseada no multiplex PCR foi comparada com os testes de difusão com disco para 1 µg/mL de oxacilina, detecção da concentração inibitória mínima (CIM), meio de triagem com 4% de NaCl e 6 µg/mL de oxacilina. Na investigação de 24 isolados obtidos de sangue, o genótipo mecA foi detectado em apenas 16 (66,7%) dos isolados pelo multiplex PCR. A CIM apresentou valores variando de 0,19 a 512 µg/mL entre os isolados de MRSA. Os dados obtidos pelos testes de triagem e diluição em ágar apresentaram sensibilidades de 80,0% e 72,8% respectivamente, quando comparados com a presença do gene mecA (multiplex). Todos os isolados, incluindo os negativos, quando reavaliados com a técnica de PCR exclusivo para este gene, o resultado foi positivo. A produção de beta-lactamase foi positiva em 20/25 (80,0%) dos isolados. Cerca de ¼ dos pacientes evoluiu para óbito apesar da maioria (83,3%) ter sido tratada adequadamente. A identificação simultânea da bactéria e sua susceptibilidade aos antibióticos é necessária para a escolha de uma terapia adequada para casos de sepse estafilocócica, mas é importante considerar a sensibilidade, especificidade e validação dos kits disponíveis.

In this study, for detection of methicillin-resistant S. aureus (MRSA), a mecA multiplex PCR-based amplification was compared with the 1 µg oxacillin disk diffusion test, detection of minimal inhibitory concentration (MIC), and screening in agar with 4% NaCl and 6 µg/mL oxacillin. Among 24 isolates obtained from blood, mecA gene was detected in only 16 (66.7%) isolates by multiplex PCR. The MIC test showed a range of resistance to oxacillin from 0.19 to 512 µg/mL, among these isolates. Data obtained by screening and dilution tests showed that sensitivity to methicillin was 80.0% and 72.8%, respectively, when compared with the presence of mecA gene (multiplex). All isolates, including the negatives, when revaluated for mecA gene by PCR were positive. beta-lactamase production was positive for 20/25 isolates (80.0%). About ¼ of patients died dispite most of them (83.3%) were adequately treated. The simultaneous identification of the bacteria and determination of this susceptibility to antibiotics are necessary for the choice of empiric antibiotic therapy in suspected staphylococcal sepse, but is important to considering the sensibility, specificity and validation of the available kits.