Orientador: Maria de Lourdes Ribeiro de Souza da Cunha / Resumo: Introdução: o isolamento de estafilococos coagulase negativos (ECN) de pacientes em Unidades de Terapia Intensiva Neonatal (UTIN) pode representar, muitas vezes, uma contaminação ao invés de bacteremia. O padrão ouro para o diagnóstico de sepse neonatal é a positividade de duas hemoculturas coletadas num intervalo de 48 horas associada às manifestações clínicas sugestivas de sepse. A resistência aos antimicrobianos e a formação de biofilme são fatores seletivos na persistência de cepas de S. epidermidis e S. haemolyticus. A técnica de Eletroforese em Gel de Campo Pulsado (PFGE), altamente discriminatória, é frequentemente usada para a investigação de surtos. A tipagem por Multilocus Sequence Typing (MLST) se tornou o método de escolha no estudo epidemiológico em longo prazo. Objetivos: esse estudo teve como objetivos caracterizar o perfil das infecções de corrente sanguínea por Staphylococcus spp. em RNs internados na UTIN, os fatores de risco para infecção, perfil de virulência e resistência antimicrobiana dos estafilococos isolados, bem como a presença de clones prevalentes na unidade. Resultados: foram isolados 72 Staphylococcus spp. de hemoculturas de 54 recém-nascidos de Março de 2014 a Agosto de 2015. Foram confirmados 37 episódios de Infecção da Corrente Sanguínea (ICS) causados por Staphylococcus spp., sendo 27 associados a espécie S. epidemidis, 3 S. haemolyticus, 2 S. capitis e 5 S. aureus. A maioria dos Recém-nascidos (RNs) possuía extremo baixo peso, nasceu com me... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Abstract Introduction: The isolation of coagulase negative staphylococci (CoNS) of Neonatal Intensive Care Unit (NICU) patients can often represent contamination instead of bacteremia. The golden standard for the diagnosis of sepsis is the positivity of two blood cultures, collected within a 48-hour gap, associated to clinic manifestations suggestive of sepsis. Resistance to antimicrobials and biofilm formation are selective factors to S. epidermidis and S. haemoyticus strains to persist. Pulsed Field Gel Electrophoresis (PFGE), a highly discriminatory technique, is often used in investigating outbreaks. Multilocus Sequence Typing (MLST) has become the preferred method when it comes long-term epidemiologic studies. Objectives: this research aimed to characterize the profile of infections caused by Staphylococcus spp. at NICU, the virulence profile and antimicrobial resistance of isolated staphylococci, as well as the presence of clusters prevalent in the unit. Results: 72 Staphyloccocus spp. have been isolated from blood cultures taken from 54 newborns from March 2014 to August 2015. Thirty-seven bloodstream infection (BSI) episodes caused by Staphylococcus spp. were confirmed, 27 of which are associated to S. epidemidis, 3 to S. haemolyticus, 2 to S. capitis and 5 to S. aureus. Most newborns presented extremely low weight, were born before the 31st week of pregnancy (72.2%), used catheter and mechanical ventilation. From the 72 samples of Staphylococcus spp. under analysis, ... (Complete abstract click electronic access below) / Doutor

Moraxella (Branhamella) catarrhalis, although commonly isolated from sputum in patients with pneumonia, rarely causes bacteremia. Sodium polyanethol sulfonate is commonly added to routine blood culture media to increase the yield of Gram-negative bacilli. This anticoagulant also inhibits the growth of Neisseria meningitidis. We added strains f Moraxella (Branhamella) catarrhalis to routine blood culture media supplemented with 0.03% SPS. No inhibitory effect could be demonstrated. The rare occurrence of bacteremic Moraxella (Branhamella) catarrhalis pneumonia is not due to an inhibitory effect of routine blood culture media.

No description available.

Introduction: Sepsis is a major source of morbidity and mortality in neonates; however, identification of the causative pathogens can be challenging. Next generation sequencing (NGS) is a high-throughput, parallel sequencing technique for DNA. Pathogen-targeted enrichment followed by NGS has the potential to be more sensitive and faster than current gold-standard blood culture. In this pilot study, we will test the feasibility and pathogen detection patterns of pathogen-targeted NGS in neonates with suspected sepsis. Additionally, the distribution and diagnostic accuracy of cell-free DNA and protein C levels at two time points will be explored. Methods: We will conduct a prospective, pilot observational study. Neonates over 1 kg with suspected sepsis from a single tertiary care children’s hospital will be recruited for the study. Recruitment will be censored at 200 events or 6 months duration. Two blood study samples will be taken: the first simultaneous to the blood culture (time = 0 hr, for NGS and biomarkers) via an exception to consent (deferred consent) and another 24 hours later after prospective consent (biomarkers only). Neonates will be adjudicated into those with clinical sepsis, culture-proven sepsis and without sepsis based on clinical criteria. Feasibility parameters (e.g. recruitment) and NGS process time will be reported. Analysis: NGS results will be described in aggregate, compared to the simultaneous blood culture (sensitivity and specificity) and reviewed via expert panel for plausibility. Pilot data for biomarker distribution and diagnostic accuracy (sensitivity and specificity) for distinguishing between septic and non-septic neonates will be reported. Study amendment and interim results: After obtaining ethics approval, study enrolment started October 15, 2020. Interim feasibility results showed successful deferred consent, but low enrolment. A study amendment was used to increase enrolment, create pre-packaged blood kits and implement a substitute decision maker Notification form. / Thesis / Master of Health Sciences (MSc)

A sepse é uma doença de alta mortalidade e o uso adequado de antimicrobianos no seu manuseio é essencial na obtenção de melhores resultados. O objetivo deste estudo foi avaliar o consumo de antimicrobianos em pacientes com sepse hospitalar com agente etiológico identificado no sangue, comparando a reação em cadeia da polimerase (PCR) - LightCycler® SeptiFast (SF) para a detecção rápida de microorganismos à hemocultura convencional nos primeiros 14 dias de tratamento. Os objetivos secundários incluíram descrever o percentual de positividade e a concordância entre o teste molecular e a hemocultura, o tempo de internação hospitalar, os custos diretos dos antimicrobianos utilizados e a letalidade em 10 e 28 dias. Entre outubro de 2012 e maio de 2016, foram incluídos 200 pacientes adultos com sepse hospitalar: 100 alocados no grupo intervenção onde a terapia antimicrobiana foi ajustada após a identificação do micro-organismo pelo SF (dentro de 6 a 12 horas) e 100 pacientes no grupo controle onde a terapia antimicrobiana foi ajustada após a identificação do microrganismo pela hemocultura (dentro de 48 a 72 horas). O consumo de antimicrobianos foi de 1429 (1071-2000) DOT por 1000 pacientes-dia no grupo intervenção versus 1889 (1357-2563) DOT por 1000 pacientes-dia no grupo controle (p = 0.017). O SF apresentou positividade de 25,9% enquanto a positividade da hemocultura foi de 22,9% (p = 0,452). O tempo de descalonamento antimicrobiano foi de 8 horas (7-14) versus 54 horas (38-75) (p < 0,001), enquanto a mortalidade em 10 e 28 dias foi de 21% e 36,8% no grupo de intervenção versus 37% e 44% no grupo controle, (p = 0,710 e p = 0,632), respectivamente. Não houve diferença no tempo de internação hospitalar e no custo direto dos antimicrobianos utilizados nos dois grupos. A duração média da terapia antimicrobiana em dias foi menor no grupo intervenção (12 ± 5 versus 15 ± 4, p = 0,039) em comparação com o grupo controle / Sepsis is a high mortality disease. Appropriate use of antimicrobials is essential to improve outcomes. The aim of the present study was to determine whether the use of the multiplex polymerase chain reaction LightCycler® SeptiFast (SF) assay reduces the antibiotic consumption through early de-escalation in patients with nosocomial sepsis compared with conventional blood cultures (BC) in the first 14 days of treatment. Secondary outcomes included the percentage of microorganisms identified through SF and BC (in both groups), timing of antimicrobial de-escalation, length of stay, direct costs of the antimicrobial drugs and mortality at 10 and 28 days. Between October 2012 and May 2016 adult patients with nosocomial sepsis were randomized in intervention group and control group: antimicrobial therapy was adjusted following the identification of microorganisms by SF (within 6 to 12 hours) or BCs (within 48 to 72 hours), respectively. A total of 200 patients were included (100 in each group). The median antimicrobial consumption was 1429 (1071-2000) DOT/1000 patients-day in the intervention group versus 1889 (1357-2563) DOT/1000 patients-day in the control (p = 0.017). Microorganism identification was 25.9% versus 22.9% (p = 0.452), timing of antimicrobial de-escalation was 8 hours (7-14) versus 54 hours (38-75) (p < 0.001) while the mortality rate at 10 and 28 days was 21% and 36.8% in the intervention group versus 37% and 44% in the control group, (p = 0.710 and p = 0.632), respectively. There was no difference in length of stay and antimicrobial costs between groups. The mean duration of antimicrobial therapy was lower in the SF group (12 ± 5 vs. 15 ± 4, p = 0.039) comparing to BC group. The use of a rapid molecular blood test resulted in a reduction in the antimicrobial consumption and a more rapid de-escalation in patients with nosocomial sepsis when compared to the standard management of blood culture

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2015-09-29T16:45:14Z No. of bitstreams: 1 Alcinéia Oliveira Damiao caracterização ....PDF.pdf: 1250268 bytes, checksum: 8e253d5ec93aa5123f3f85b88d8bc154 (MD5) / Approved for entry into archive by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2015-09-29T16:45:29Z (GMT) No. of bitstreams: 1 Alcinéia Oliveira Damiao caracterização ....PDF.pdf: 1250268 bytes, checksum: 8e253d5ec93aa5123f3f85b88d8bc154 (MD5) / Made available in DSpace on 2015-09-29T16:45:29Z (GMT). No. of bitstreams: 1 Alcinéia Oliveira Damiao caracterização ....PDF.pdf: 1250268 bytes, checksum: 8e253d5ec93aa5123f3f85b88d8bc154 (MD5) Previous issue date: 2015 / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / A leptospirose é uma zoonose de distribuição mundial, com 1,2 milhões de casos registrados a cada ano. De 1996 a 2013, o grupo de pesquisa de leptospirose do CPqGM, realiza uma vigilância ativa no Hospital Couto Maia em Salvador-Ba, onde foi recrutado 4612 casos suspeitos para leptospirose. Destes 4612 foi confirmado o diagnóstico de 1853 (40%) utilizando pelo menos um dos três métodos de diagnóstico (MAT, Hemocultura, qPCR). Dentre os casos confirmados, 1759 (95%) foram confirmados pelo MAT. A sensibilidade do MAT foi diferente entre as amostras aguda e convalescente, sendo 60% na fase aguda e 97% na fase convalescente. O sorogrupo Icterohaemorrhagiae foi o mais prevalente (90%) dos casos confirmados para MAT. Durante o período do estudo foram coletadas 1133 hemoculturas e destas 203 (18%) foram positivas, sendo possível isolar leptospiras de 93/203 (45%) das hemoculturas, as quais foram soro-agrupadas com soros heterológos de coelho. A concordância entre o sorogrupo encontrado no MAT e na soro-agrupagem foi de 80%. Os achados mostram que existe uma concordância significante entre o sorogrupo encontrado pelos dois métodos, o que indica que o painel de cepas utilizado no MAT apresenta uma ótima cobertura para os sorogrupos prevalentes na região. A predominância de um sorogrupo facilitou quanto a tomadas de decisões para prevenção e controle, assim como facilita para o desenvolvimento de novos testes de diagnóstico e vacinas mais direcionados. / Leptospirosis is a re-emerging zoonotic disease of global importance, with 1,2 million cases reported each year. Diagnosis of leptospirosis is often difficult given the nonspecific disease presentation. In order to compare the performance of the two gold standard diagnostic tests for leptospirosis, the group enrolled 4612 patients with suspected leptospirosis during active surveillance at the state infectious disease reference hospital, Hospital Couto Maia, in Salvador, Bahia between 1996 and 2013. Of these, was confirmed Leptospira infection in 1853 (40%) using at least one of three diagnostic methods (microagglutination (MAT), blood culture, and qPCR). Was confirmed 1759 (95%) cases using only the MAT assay, and identified the serogroup Icterohaemorrhagiae as the infective agent in 90% of MAT positive samples. It was determined the sensitivity of the MAT was 60% for acute phase samples and increased to 97% for convalescent samples. Within this study period, it was possible to collect 1133 blood cultures and was isolated leptospires from 93 of 203 (45%) of blood cultures, and determined the serotype using heterologous rabbit sera. The concordance between the infective serogroup identified using hemoculture and MAT techniques was 80%. This result indicates that the panel of 11 strains used in the MAT represents a majority of the infective serogroups causing disease in our study population. The predominance of a single serogroup in symptomatic cases informs the development of new diagnostic tests and novel vaccines to prevent leptospirosis in Brazil.

A sepse é uma doença de alta mortalidade e o uso adequado de antimicrobianos no seu manuseio é essencial na obtenção de melhores resultados. O objetivo deste estudo foi avaliar o consumo de antimicrobianos em pacientes com sepse hospitalar com agente etiológico identificado no sangue, comparando a reação em cadeia da polimerase (PCR) - LightCycler® SeptiFast (SF) para a detecção rápida de microorganismos à hemocultura convencional nos primeiros 14 dias de tratamento. Os objetivos secundários incluíram descrever o percentual de positividade e a concordância entre o teste molecular e a hemocultura, o tempo de internação hospitalar, os custos diretos dos antimicrobianos utilizados e a letalidade em 10 e 28 dias. Entre outubro de 2012 e maio de 2016, foram incluídos 200 pacientes adultos com sepse hospitalar: 100 alocados no grupo intervenção onde a terapia antimicrobiana foi ajustada após a identificação do micro-organismo pelo SF (dentro de 6 a 12 horas) e 100 pacientes no grupo controle onde a terapia antimicrobiana foi ajustada após a identificação do microrganismo pela hemocultura (dentro de 48 a 72 horas). O consumo de antimicrobianos foi de 1429 (1071-2000) DOT por 1000 pacientes-dia no grupo intervenção versus 1889 (1357-2563) DOT por 1000 pacientes-dia no grupo controle (p = 0.017). O SF apresentou positividade de 25,9% enquanto a positividade da hemocultura foi de 22,9% (p = 0,452). O tempo de descalonamento antimicrobiano foi de 8 horas (7-14) versus 54 horas (38-75) (p < 0,001), enquanto a mortalidade em 10 e 28 dias foi de 21% e 36,8% no grupo de intervenção versus 37% e 44% no grupo controle, (p = 0,710 e p = 0,632), respectivamente. Não houve diferença no tempo de internação hospitalar e no custo direto dos antimicrobianos utilizados nos dois grupos. A duração média da terapia antimicrobiana em dias foi menor no grupo intervenção (12 ± 5 versus 15 ± 4, p = 0,039) em comparação com o grupo controle / Sepsis is a high mortality disease. Appropriate use of antimicrobials is essential to improve outcomes. The aim of the present study was to determine whether the use of the multiplex polymerase chain reaction LightCycler® SeptiFast (SF) assay reduces the antibiotic consumption through early de-escalation in patients with nosocomial sepsis compared with conventional blood cultures (BC) in the first 14 days of treatment. Secondary outcomes included the percentage of microorganisms identified through SF and BC (in both groups), timing of antimicrobial de-escalation, length of stay, direct costs of the antimicrobial drugs and mortality at 10 and 28 days. Between October 2012 and May 2016 adult patients with nosocomial sepsis were randomized in intervention group and control group: antimicrobial therapy was adjusted following the identification of microorganisms by SF (within 6 to 12 hours) or BCs (within 48 to 72 hours), respectively. A total of 200 patients were included (100 in each group). The median antimicrobial consumption was 1429 (1071-2000) DOT/1000 patients-day in the intervention group versus 1889 (1357-2563) DOT/1000 patients-day in the control (p = 0.017). Microorganism identification was 25.9% versus 22.9% (p = 0.452), timing of antimicrobial de-escalation was 8 hours (7-14) versus 54 hours (38-75) (p < 0.001) while the mortality rate at 10 and 28 days was 21% and 36.8% in the intervention group versus 37% and 44% in the control group, (p = 0.710 and p = 0.632), respectively. There was no difference in length of stay and antimicrobial costs between groups. The mean duration of antimicrobial therapy was lower in the SF group (12 ± 5 vs. 15 ± 4, p = 0.039) comparing to BC group. The use of a rapid molecular blood test resulted in a reduction in the antimicrobial consumption and a more rapid de-escalation in patients with nosocomial sepsis when compared to the standard management of blood culture

Contaminated blood cultures constitute diagnostic challenges and place a burden on healthcare services. An observational retrospective study was undertaken to evaluate the effect of routine labelling of blood culture bottles with the initials of the healthcare worker who drew them, followed by individualized feedback, on blood culture contamination rates. The contamination rate of the entire facility was 2.6% before the procedural change, and this decreased significantly to 1.5% after the procedural change (P < 0.001) over the first 12 months of the intervention. Routine labelling of blood culture bottles with the initials of the healthcare worker who drew them, followed by individualized feedback, was effective in reducing blood culture contamination rates.

<p>The aim of this study was to investigate the effects of Sutherlandia frutescens on the inflammatory response and T cell differentiation in vitro using cytokines as biomarkers. Whole blood cells containing various concentrations of Sutherlandia frutescens were stimulated in vitro with either Lipopolysaccharide (LPS) or Phytohaemagglutinin (PHA). Results show that Sutherlandia frutescens is not toxic at any of the concentrations tested. The addition of Sutherlandia frutescens at high concentrations to the stimulated whole blood cell cultures reflects a significant down regulation of Interleukin(IL) 6 and IL-10 compared to the control (P&lt / 0.05) hence suppressed the inflammatory and humoral immune response. Results obtained for Inteferon-gamma (IFN ) shows that Sutherlandia frutescens is donor specific as it reflects both up and down regulation in the release of IFN at the concentrations tested. The in vitro data generated by this study supports the use of Sutherlandia frutescens in the management of inflammatory conditions and allergies such as asthma. However the effects of Sutherlandia frutescens on cell mediated immunity was found to be donor specific. Further investigation of Sutherlandia frutescens on cellular immunity is advised.</p>

<p>The aim of this study was to investigate the effects of Sutherlandia frutescens on the inflammatory response and T cell differentiation in vitro using cytokines as biomarkers. Whole blood cells containing various concentrations of Sutherlandia frutescens were stimulated in vitro with either Lipopolysaccharide (LPS) or Phytohaemagglutinin (PHA). Results show that Sutherlandia frutescens is not toxic at any of the concentrations tested. The addition of Sutherlandia frutescens at high concentrations to the stimulated whole blood cell cultures reflects a significant down regulation of Interleukin(IL) 6 and IL-10 compared to the control (P&lt / 0.05) hence suppressed the inflammatory and humoral immune response. Results obtained for Inteferon-gamma (IFN ) shows that Sutherlandia frutescens is donor specific as it reflects both up and down regulation in the release of IFN at the concentrations tested. The in vitro data generated by this study supports the use of Sutherlandia frutescens in the management of inflammatory conditions and allergies such as asthma. However the effects of Sutherlandia frutescens on cell mediated immunity was found to be donor specific. Further investigation of Sutherlandia frutescens on cellular immunity is advised.</p>