Biologia molecular aplicada ao diagnóstico laboratorial de endoftalmite bacteriana

Bacterial endophthalmitis is a serious but uncommon intraocular infection which frequently results in vision loss. Early diagnosis and appropriate therapy are associated with better visual outcome. Conventional microbiological methods are currently used for microbiological characterization of eyes with suspected endophthalmitis. However, the sensitivity of bacterial detection from aqueous and vitreous humor using microbiology techniques is poor, and time-consuming to confirm the results. The application of molecular methods enhances significantly laboratory confirmation of bacterial endophthalmitis, demanding a shorter time to draw a definitive result and thereby promoting the early initiation of a more specific therapy to limit the empirical or unnecessary use of broad-spectrum antibiotics. PCR-based techniques, including post-PCR methods such RFLP, DNA probe hybridization and DNA sequencing have been successfully used for the diagnostic elucidation of clinically suspected bacterial endophthalmitis cases, showing promising application in the routine practice of ocular microbiology laboratories.Endoftalmite bacteriana é uma infecção intraocular grave, mas de baixa frequência, podendo resultar em grande prejuízo visual. O diagnóstico precoce e a rápida instituição de terapia adequada estão associadas a um melhor prognóstico da doença. Os métodos microbiológicos convencionais são utilizados rotineiramente para caracterização microbiológica de olhos com suspeita de endoftalmite. No entanto, a sensibilidade de detecção bacteriana em amostras de humor aquoso e vítreo utilizando técnicas microbiológicas é baixa, além de demandar um maior tempo para a confirmação dos resultados. A utilização de métodos moleculares aumenta significativamente os casos de endoftalmite bacteriana confirmados laboratorialmente, com tempo menor para a liberação de um resultado definitivo, auxiliando assim a instituição precoce de uma terapia mais específica, limitando o uso empírico ou desnecessário de antibióticos de amplo espectro. A técnica de PCR e outras metodologias para análises pós-PCR como, RFLP, hibridização com sondas e sequenciamento do DNA, tem sido utilizadas com sucesso para elucidação diagnóstica em casos com suspeita clínica de endoftalmite bacteriana, demonstrando promissora aplicação para a rotina dos laboratórios de microbiologia ocular.Universidade Federal de São Paulo (UNIFESP) Departamento de Medicina Laboratório ALERTAUNIFESP Departamento de OftalmologiaUNIFESP Laboratório Especial de Microbiologia ClínicaUNIFESP, Depto. de Medicina Laboratório ALERTAUNIFESP, Depto. de OftalmologiaUNIFESP, Laboratório Especial de Microbiologia ClínicaSciEL

Prevalência e suscetibilidade antibiótica de Staphylococcus aureus resistente à meticilina em infecções oculares

PURPOSE: To study the prevalence of methicillin-resistant Staphylococcus aureus among S. aureus ocular infections in a tertiary health center in Brazil and compare antibiotic susceptibility patterns between MRSA and methicillin-susceptible S. aureus isolates. METHODS: Electronic records from the ocular microbiology laboratory of the Universidade Federal de São Paulo (UNIFESP) were retrospectively reviewed. During a 10-year period (between January 2000 and December 2009) all conjunctivitis, keratitis, and endophthalmitis cases with a positive culture for S. aureus were identified. Antibiotic susceptibility was determined using the Kirby-Bauer disk diffusion method. RESULTS: Five hundred sixty-six S. aureus isolates were identified; of those, 56 (9.9%) were resistant to methicillin. Throughout the 10-year period, Staphylococcus aureus showed a significant increasing trend from 7.55% to 16.18% among overall S. aurues infections (p=0.001) and from 3.7% to 13.16% in conjunctivitis (p=0.001). Conversely, we did not observe the same trend among those with keratitis (p=0.38). Staphylococcus aureus isolates showed higher resistance rates to tobramycin, gentamicin, ciprofloxacin, gatifloxacin, and moxifloxacin when compared with S. aureus isolates (p< 0.001). All cases were susceptible to vancomycin. CONCLUSION: We observed an increasing trend in the overall prevalence of Staphylococcus aureus ocular infections and statistically significant higher resistance rates to commonly used antibiotics compared to Staphylococcus aureus. Our data supports the need for constant bacterial surveillance and should be taken into consideration before initiating empiric treatment of ocular infections.OBJETIVO: Estudar a prevalência do Staphylococcus aureus resistente à meticilina nas infecções oculares causadas por S. aureus em um centro de saúde terciário no Brasil e comparar o perfil de suscetibilidade antimicrobiana entre as cepas de Staphylococcus aureus resistente à meticilina e S. aureus susceptível à meticilina MÉTODOS: Foi realizada uma análise retrospectiva dos arquivos do laboratório de microbiologia ocular da Universidade Federal de São Paulo (UNIFESP) e selecionados todos os casos de conjuntivite, ceratite e endoftalmite com cultivo positivo para S. aureus, durante um período de 10 anos (entre janeiro de 2000 e dezembro de 2009). Foi avaliada a prevalência de Staphylococcus aureus resistente à meticilina e comparado o perfil de susceptibilidade antimicrobiano dos Staphylococcus aureus resistente à meticilina e S. aureus susceptível à meticilina. RESULTADOS: Quinhentos e sessenta e seis isolados de S. aureus foram identificados. Desses, 56 (9,9%) apresentaram resistência à meticilina. Durante o período de 10 anos estudado, Staphylococcus aureus resistente à meticilina mostrou uma tendência significante de aumento de 7,6% para 16,2% entre as infecções oculares causadas por S. aureus em geral (p=0,001) e de 3,7% para 13,2% nas conjuntivites (p=0,001). A mesma tendência não foi observada entre as amostras de ceratite (p=0,38). Os isolados de Staphylococcus aureus resistente à meticilina mostraram maiores taxas de resistência à tobramicina, gentamicina, ciprofloxacino, gatifloxacino e moxifloxacino em comparação com os isolados de S. aureus susceptível à meticilina (p<0,001). Todos os casos foram susceptíveis à vancomicina. CONCLUSÃO:Foi observada uma tendência de aumento na prevalência do Staphylococcus aureus resistente à meticilina nas infecções oculares causadas por S. aureus, bem como taxas de resistência significantemente maiores aos antibióticos comumente utilizados na prática oftalmológica. Nossos dados alertam para a necessidade de constante vigilância de resistência bacteriana a antimicrobianos e devem ser considerados na eleição do tratamento empírico das infecções oculares.Universidade Federal de São Paulo (UNIFESP) Department of OphthalmologyUniversidad de la República Facultad de Medicina Instituto de HigieneUNIFESP, Department of OphthalmologySciEL

Culture proven bacterial endophthalmitis: a 6-year review

PURPOSE: To assess the distribution of microorganisms isolated from patients with bacterial endophthalmitis and their antimicrobial susceptibility. METHODS: Retrospective analysis of medical and microbiological records of patients with suspected diagnosis of endophthalmitis and bacterial culture-proven at the Department of Ophthalmology, UNIFESP, between January 1 2000 and December 31 2005. RESULTS: 153 (33.9%) of 451 patients showed positive bacterial culture. A total of 155 microorganisms were isolated, 79.35% were gram-positive and 20.65% gram-negative. Staphylococcus (CoNS) (41.94%) were the most frequently isolated. The antimicrobial susceptibility for gram-negative microorganisms was as follows: amikacin 87.10%, tobramycin 80.65%, ciprofloxacin 96.67%, levofloxacin, gatifloxacin and moxifloxacin 100%, ceftazidime 85.0%, and gentamicin 80.65%. Vancomycin sensitivity among gram-positive microorganisms was 100%. S. aureus and CoNS showed 83.33% of susceptibility to oxacillin, 89.61% to ciprofloxacin and 100% to gatifloxacin and moxifloxacin. The main acquisition mechanism was postoperative (60.65%). CONCLUSION: We detected a low sensitivity of vitreous/aqueous culture for the etiologic diagnosis of endophthalmitis. The empiric antimicrobial therapy or prophylaxis should be active against gram-positive bacteria, particularly staphylococci. Surveillance studies of bacterial resistance are important for a better utilization of antimicrobials in this clinical setting.OBJETIVO: Determinar a distribuição dos microrganismos isolados de pacientes com endoftalmite bacteriana e sua sensibilidade a antimicrobianos. MÉTODOS: Foram analisados retrospectivamente os dados clínicos e microbiológicos dos pacientes com hipótese diagnóstica de endoftalmite e cultura bacteriana positiva, atendidos no Departamento de Oftalmologia da UNIFESP de 1º de janeiro de 2000 a 31 de dezembro de 2005. RESULTADOS: De 451 pacientes, 153 (33,9%) apresentaram cultura bacteriana positiva. Foram isolados 155 microrganismos, sendo 79,35% gram-positivos e 20,65% gram-negativos. Os Staphylococcus coagulase-negativos (SCoN) (41,94%) foram os mais freqüentemente isolados. A sensibilidade aos antimicrobianos entre os gram-negativos foi: amicacina 87,10%, tobramicina 80,65%, ciprofloxacina 96,67%, levofloxacina, gatifloxacina e moxifloxacina 100%, ceftazidima 85%, e gentamicina 80,65%. A sensibilidade à vancomicina entre os gram-positivos foi de 100%. S. aureus e SCoN apresentaram 83,33% de sensibilidade à oxacilina, 89,61% à ciprofloxacina e 100% à gatifloxacina e moxifloxacina. A forma de aquisição predominante foi a pós-operatória (60,65%). CONCLUSÃO: Observamos baixa sensibilidade da cultura para o diagnóstico etiológico das endoftalmites. Uma terapia antimicrobiana ou profilaxia empírica deve ser ativa contra os microrganismos gram-positivos, particularmente contra estafilococos. Estudos de vigilância de resistência bacteriana são importantes para adequação desses esquemas.Universidade Federal de São Paulo (UNIFESP) - UNIFESP Departamento de Medicina Laboratório Especial de Microbiologia Clínica - LEMCUNIFESP Departamento de OftalmologiaFundação Faculdade Federal de Ciências Médicas de Porto Alegre - FFFCMPA Departamento de Microbiologia e ParasitologiaUNIFESP Departamento de Oftalmologia Laboratório de MicrobiologiaUNIFESP Departamento de Oftalmologia Laboratório de Microbiologia OcularUNIFESP Departamento de Medicina Laboratório Especial de Microbiologia ClínicaUNIFESP, Depto. de Medicina Laboratório Especial de Microbiologia Clínica - LEMCUNIFESP, Depto. de OftalmologiaUNIFESP, Depto. de Oftalmologia Laboratório de MicrobiologiaUNIFESP, Depto. de Oftalmologia Laboratório de Microbiologia OcularUNIFESP, Depto. de Medicina Laboratório Especial de Microbiologia ClínicaSciEL

Incidência de endoftalmite após cirurgia de catarata (2002-2008) num hospital universitário brasileiro

Purpose: To report on the incidence, diagnostic technique, and microbiological features of endophthalmitis at a university-setting in Brazil. Methods: All cases of presumed postoperative endophthalmitis from 2002 to 2008 at a teaching-hospital were included. Main data assessed were: number of cataract surgeries performed, incidence of endophthalmitis, microbiological outcome (aqueous and/or vitreous culture and Gram staining), and antimicrobial susceptibility testing of the positive cases. Results: Seventy-three eyes of 73 patients (43 females and 30 males) developed endophthalmitis after 24,590 cataract surgeries. The incidence decreased from 0.49% in 2003 to 0.17% in 2006 and stabilized afterwards. Coagulase negative Staphylococci (CoNS) and Streptococcus viridans (56.5% and 15%, respectively) were the most common bacterial isolates. Culture and Gram stain were negative in 36.9%. CoNS presented susceptibility rates of 80%-sensitivity to oxacillin, 90% to fourth-generation quinolones and 100% to vancomycin. Conclusions: The rate of endophthalmitis, diagnostic ability of conventional laboratory investigation, microbial isolates and antibiotic susceptibility are in accordance with other findings of the literature. Despite using prophylactic antibiotic drops, it was possible to identify cases that were susceptible to the antibiotics topically applied.Objetivo: Relatar incidência, técnica diagnóstica e características microbiológicas de endoftalmite numa instituição universitária no Brasil. Métodos: Todos os casos de endoftalmite pós-operatória presumida de 2002 a 2008 foram incluídos. Os principais dados avaliados foram: número de cirurgias de catarata realizadas, incidência de endoftalmite, resultado microbiológico (bacterioscopia e cultura de aquoso e vítreo) e teste de sensibilidade antibiótica dos casos positivos. Resultados: Setenta e três olhos de 73 pacientes (43 do sexo feminino e 30 do masculino) desenvolveram endoftalmite após 24.590 cirurgias de catarata. A incidência reduziu de 0,49% em 2003 para 0,17% em 2006 e estabilizou-se depois disso. Staphylococcus coagulase-negativa (SCoN) e Streptococcus viridans (56.5% e 15%, respectivamente) foram os isolados bacterianos mais comuns. Cultura e bacterioscopia foram negativas em 36,9%. SCoN apresentou taxas de sensibilidade de 80% à oxacilina, 90% às quinolonas de quarta geração e 100% à vancomicina. Conclusões: A taxa de endoftalmite, a capacidade diagnóstica das técnicas laboratoriais convencionais, os microrganismos isolados e a sensibilidade aos antibióticos estão em acordo com outros achados na literatura. Apesar do uso profilático de colírio antibiótico, foi possível identificar casos de infecção em que as bactérias eram sensíveis aos antibióticos usados topicamente.Universidade Federal de São Paulo (UNIFESP) Department of OphthalmologyUniversidade Federal de São Paulo (UNIFESP) Department of Medicine Special Laboratory of Clinical MicrobiologyUniversidade Federal de São Paulo (UNIFESP) Ocular Microbiology LaboratoryUNIFESP, Department of OphthalmologyUNIFESP, Department of Medicine Special Laboratory of Clinical MicrobiologyUNIFESP, Ocular Microbiology LaboratorySciEL

Influence of feeding graded levels of canned sardines on the inflammatory markers and tissue fatty acid composition of Wistar rats

The present study was supported by Fundacao para a Ciencia e a Tecnologia (FCT) through the grant PTDC-SAU/OSM/2006/70560 (principal investigator: P. O. R). S. V. M. is the recipient of a FCT individual fellowship SFRH/BPD/2009/63019 and P. A. L. is a researcher involved in the FCT programme 'Ciencia 2008'.Canned sardines are a ready-to-use fish product with excellent nutritional properties owing to its high n-3 long-chain PUFA content, mainly EPA (20 : 5n-3) and DHA (22 : 6n-3). The present study aimed to assess the effect of two dosages of canned sardines, recommended for the primary and secondary prevention of human CVD, on the inflammatory marker concentrations and fatty acid composition of erythrocytes and key metabolic tissues (liver, muscle, adipose tissue and brain) in the rat model. Wistar rats were fed a diet containing 11\% (w/w) of canned sardines (low-sardine (LS) diet) and a diet containing 22\% (w/w) of canned sardines (high-sardine (HS) diet) for 10 weeks. Daily food intake, weight gain, and organ and final body weights were not affected by the dietary treatments. The concentrations of total cholesterol, HDL-cholesterol and LDL-cholesterol decreased in both the LS and HS groups, while those of alanine aminotransferase and adiponectin increased. The concentrations of IL-1 beta increased only with the highest dosage of sardine. The dose-dependent influence of the graded levels of EPA+DHA was tissue specific. Compared with that of other tissues and erythrocytes, the fatty acid composition of the brain was less affected by the canned sardine-supplemented diets. In contrast, the retroperitoneal adipose tissue was highly responsive. The deposition ratios of EPA and DHA indicated that the LS diet was optimal for DHA deposition across the tissues, except in the retroperitoneal adipose tissue. Taken together, our findings indicate that a LS diet positively affects plasma lipid profiles and inflammatory mediators, whereas a HS diet has contradictory effects on IL-1 beta, which, in turn, is not associated with variations in the concentrations of other pro-inflammatory cytokines. This finding requires further investigation and pathophysiological understanding.publishersversionpublishe

Butyrate Protects Mice from Clostridium difficile-Induced Colitis through an HIF-1-Dependent Mechanism

Antibiotic-induced dysbiosis is a key factor predisposing intestinal infection by Clostridium difficile. Here, we show that interventions that restore butyrate intestinal levels mitigate clinical and pathological features of C. difficile-induced colitis. Butyrate has no effect on C. difficile colonization or toxin production. However, it attenuates intestinal inflammation and improves intestinal barrier function in infected mice, as shown by reduced intestinal epithelial permeability and bacterial translocation, effects associated with the increased expression of components of intestinal epithelial cell tight junctions. Activation of the transcription factor HIF-1 in intestinal epithelial cells exerts a protective effect in C. difficile-induced colitis, and it is required for butyrate effects. We conclude that butyrate protects intestinal epithelial cells from damage caused by C. difficile toxins via the stabilization of HIF-1, mitigating local inflammatory response and systemic consequences of the infection

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio