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Bakgrund: Enterohemorragisk Escherichia coli (EHEC) är en toxinproducerande bakterie somkan orsaka sporadiska fall av infektion men även ge upphov till allvarliga utbrott. Sjukdomen kan ge symtom alltifrån okomplicerad diarré, blodig diarrétill mycket allvarliga symtomsom hemolytiskt uremiskt syndrom (HUS). De flesta EHEC-falli Sverige förekommer i åldersgruppen ett till fyra år. Syfte: Studienssyfte var att undersöka värdet av en EHEC-screening hos barn under tio år genom att kartlägga förekomst av EHEC och distribution av serotypoch stx-typer. Ytterligare ett syfte var att undersöka om den kliniska bilden är beroende av serotyp och stx-typ. Därutöver undersöktes hur länge man utsöndrars tx i avföringen. Metod: I studien ingick alla barn under tio års ålder, som lämnat avföringsprov under perioden 1 maj 2003 till 30 april 2013i Jönköpingslän. Barnen delades in i de med klinisk EHEC frågeställning och de resterande som ingick i screeningen. Barnen följdes med upprepad provtagning varje vecka efter första positiva provet. Kliniska data samlades in via ett frågeformulär samt via granskning av journaler. Resultat: Totalt diagnostiserades 191 barn (87 flickor och 104 pojkar) med EHEC, av dessa var 162 indexfall och 29 hittades vid smittspårning. Prevalensen av EHEC var 1,8% i gruppen med klinisk frågeställning och 1,5% ingick i screening (p=0,5). Mediantiden för utsöndringav stx i avföringen var 20 dagar (1-256 dagar). Barnmed Stx2 producerande EHEC hade allvarligare symtom. Sju barn insjuknade i HUS, alla var smittade i Sverige. Det var vanligare med Stx2 hos barn smittade i Sverige. I fem av fallen kunde smittkällan fastställas. Konklusion: Studienvisar en lika högprevalens av EHEC i den screenade gruppen som i gruppen där EHEC var efterfrågat. Även svårighetsgraden av symtom var lika i båda grupperna.EHEC-förekomstenvar hög och de flestavarejav serotyp O157. Serotypoberoende metoder vid diagnostikär därför viktiga. Den långa utsöndringstiden utgör en risk för smittspridning. De flesta barnen med Stx2 var smittade i Sverige, vilket medför ökad risk för svår sjukdom. Studiens resultatbekräftar värdet av EHEC-screening hos barn. / Background:Enterohemorrhagic Escherichia coli(EHEC) is a toxin-producing bacterium responsible for sporadic cases of infection as well as serious outbreaks. Symptoms rangefrom uncomplicated or bloody diarrhea to hemolytic uremic syndrome (HUS). In Sweden, most EHEC casesoccurin 1-4-year-old children. Aims:This study aimed at investigate the value of EHEC screening in children younger than 10 years of age by evaluatingthe prevalence of EHEC and the distribution of serotypes and stxtypes. We also aimed t ocorrelate clinical symptoms with EHEC serotypes and stxtypes. Furthermore the duration of faecal shedding of stxwas investigated. Methods: The study examined stool samples collected fromall children younger than 10 years of age between1 May 2003 and April 2013 in the County of Jönköping. We divided the children into a physician-requested EHEC analysis group and ascreening group. Children who tested positive for stxwere sampled weekly after initial EHEC diagnosis. We used a questionnaire and reviewed medical records to collect clinical data. Results: Among 191 children (87 girls and 104 boys) with confirmed EHEC, 162 specimens were index cases and 29 were found by contact tracing. The EHEC prevalence was 1.8 % in the EHEC requested group and 1.5 % in the screening group (p=0.5). The median duration of stxdetection in faeces was 20 days (1-256 days). Symptoms were more severe in children with Stx2-producing EHEC, and seven children developed HUS, all infected in Sweden. We were able to determine the source of infection in five cases. Conclusions: This study showed that the EHEC prevalence and severity of symptoms where equal between the requested and screening group. HighEHEC prevalence and a high proportion of non-O157 isolates were found. Our results emphasize the need for serotype-independent methods for EHEC detection. The long duration of stxdetection in stools indicates a risk for transmission. Most children with Stx2 were infected in Sweden, suggesting a higher risk forsevere symptoms. Our results confirm the value of EHEC screening in children. / <p>ISBN 978-94-86739-92-8</p>

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2012-07-25T20:57:02Z No. of bitstreams: 1 Luis da Conseiçao Martins Morais Epidemiologia molecular de Streptococcus pneumoniaes... 2012.pdf: 2694690 bytes, checksum: 6a8757e0acddf633d2db80ca6ea13e0c (MD5) / Made available in DSpace on 2012-07-25T20:57:02Z (GMT). No. of bitstreams: 1 Luis da Conseiçao Martins Morais Epidemiologia molecular de Streptococcus pneumoniaes... 2012.pdf: 2694690 bytes, checksum: 6a8757e0acddf633d2db80ca6ea13e0c (MD5) Previous issue date: 2012 / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, Brasil / Streptococcus pneumoniae é a principal causa de morbi-mortalidade no mundo. Em Moçambique, os sorotipos 1 e 5 são os mais prevalentes. Este estudo investigou a relação clonal de isolados de pneumococo obtidos de doença invasiva entre o período de 2002-2007, utilizando três procedimentos laboratoriais: Box-PCR, PFGE e MLST. Um total de 105 isolados (sendo 72 do sorotipo 1 e 33 do sorotipo 5), foram submetidos a técnica de Box-PCR. O sorotipo 5 apresentou cinco padrões, sendo um clonal com 20 isolados e quatro padrões não clonais. O padrão A foi o predominante com 61% dos isolados, enquanto que por PFGE, 100% dos isolados foram agrupados em um único clone (clone A), e pela técnica de MLST foi identificado apenas um único ST (ST 289). Por outro lado, o sorotipo 1 apresentou maior diversidade clonal pelos três métodos; por Box-PCR os isolados foram agrupados em 3 clones, sendo predominante o padrão B com 58 isolados, padrão C e N com dois isolados cada. Um total de 12 isolados foram não clonais. O clone B apresentou 20 subtipos, sendo o mais frequente o sub-tipo B1 com 20 amostras idênticas, seguido por B2, B6, B7 com 5 amostras idênticas cada. Por PFGE, 19 amostras confirmaram ser do mesmo perfil clonal (clone B), enquanto que 12 amostras demonstraram ser não clonais. Quando submetidos ao MLST, foram identificados 6 STs; ST 217 (7 isolados), ST 853 (1 isolado), e quatro novos STs, ST 4125, ST 2909, ST 3779 e ST 4166. O ST 217 pertence ao clone Suécia1-27 (ST217), identificados previamente em surtos de meningite na África, enquanto o ST 289 foi identificado como um representante do clone Colômbia5-19 (ST289) que circulam na América Latina desde 1994. A taxa de não susceptibilidade à penicilina foi de 3%, e à cotrimoxazole foi de 39%. A maior taxa de resistência foi encontrada entre os isolados de sorotipo 1. Este trabalho mostra a persistência de dois sorotipos responsáveis por causar doença pneumocócica invasiva graves, bem como seus respectivos clones em uma região do sul de Moçambique. / Streptococcus pneumoniae is the leading cause of morbidity and mortality worldwide. In Mozambique, serotypes 1 and 5 are the most prevalent. This study investigated the clonal relationship of isolates obtained from pneumococcal invasive disease during the period of 2002-2007 using three laboratory procedures: Box-PCR, PFGE and MLST. A total of 105 isolates (72 serotype 1 and 33 serotype 5) were submited to Box-PCR technique. The serotype 5 showed five patterns, one clonal with 20 isolates and four non-clonal patterns. The pattern A, covered 61% of all isolates, whereas by PFGE, 100% of the isolates have been grouped into a single clone (clone A), and by MLST was also identified only one ST (ST 289). In another hand, serotype 1 had higher clonal diversity by the three methods; by Box-PCR isolates have been grouped into three clonal patterns, the pattern B being predominantly (58 isolates), C and N with two isolates each. A total of 12 strains were non-clonal. Clone B showed 20 sub-types, the most common subtype B1 with 20 identical samples, followed by B2, B6, B7 with 5 identical samples each. By PFGE, 19 samples confirmed with the same profile clone (clone B), while 12 samples were non-clonal. By MLST, 6 STs were identified, ST 217 (7 isolates), ST 853 (one isolate) and four new STs, ST 4125, ST 2909, ST 3779 and ST 4166. ST 217 belongs to Sweden1-27 clone (ST217), previously identified in outbreaks of meningitis in Africa, while the ST 289 has been identified as a representative clone Colômbia5-19 clone (ST289) that circulate in Latin America since 1994. The rate of non susceptibility to penicilin was 3%, and 39% to cotrimoxazol. The highest resistance rate was found among serotype 1 isolates. This results shows the persistence of two serotypes responsible for cases of severe invasive pneumococcal disease, as well as their respective clones in a region of southern Mozambique.

The diagnosis of dengue cases in Central Laboratories of Public Health (LACEN) of the country includes the research of NS1 protein and only in positive cases, diagnostic confirmation occurs by identifying the serotype. Data provided by the Epidemiological Surveillance of Sergipe revealed that the first half of 2016 all suspected cases that reached the LACEN / SE were negative on screening tests, even in the thirty-first epidemiological week of the same year, Sergipe has reached the seventh place in the racking of dengue cases in the Northeast region and the third in the Quick Survey Infestation Index by Aedes aegypti. This study aimed to identify the serotypes and the epidemiological profile of suspected dengue cases in the first half of 2016. The 437 suspected dengue patients from January to June of this year were registered in LACEN / SE. After exclusion criteria, 382 serum samples of these patients diagnosed and negative for dengue by ELISA NS1 Antigen Platelia kit method were analyzed by RT-PCR in real time on the multiplex system for confirmation of suspected cases of dengue. The distribution of patients according to epidemiological and demographic variables revealed that there was a predominance of cases in the south central region of the state and large Aracaju (53.5%) inferring the ease in access to outpatient and laboratory service in the region the most affected gender was female (62.8%) and the age group of highest incidence was 20-59 years (59.3%). The highest rate was in urban areas (84.9%) and the first three days of symptoms (63.9%) with higher sampling rate. The prevalence of dengue in cases initially presented as a suspect was 22.5% (86) distributed among serotypes DENV4 82.5% (71), DENV1 9.3% (8) and DENV3 8.1% (7). Compared to official data reported that there were no cases of dengue in the period, this study reveals the positive for dengue with cocirculation of three distinct serotypes being DEV4 the predominant. As also, reports the first evidence of the last ten years of DENV3 serotype circulating in the state. Data analysis enabled to view an underreporting of cases screened and thus suggest a reassessment of the methods used as diagnostic screening so that they can take control measures to the potential for severe disease in a population. / O diagnóstico de casos de dengue nos Laboratórios Centrais de Saúde Pública (LACEN) do país contempla a pesquisa da proteína NS1 e, somente em casos positivos, a confirmação diagnóstica ocorre pela identificação do sorotipo. Dados disponibilizados pela Vigilância epidemiológica de Sergipe revelaram que no primeiro semestre de 2016 todos os casos suspeitos que chegaram ao LACEN/SE foram considerados negativos nos testes de triagem, ainda que na trigésima primeira semana epidemiológica do mesmo ano, Sergipe tenha alcançado o sétimo lugar no racking de casos de dengue da região Nordeste e o terceiro no Levantamento Rápido do Índice de Infestação por Aedes aegypti. O presente trabalho teve como objetivo conhecer da dengue no estado de Sergipe no primeiro semestre de 2016 relacionando-os com alguns parâmetros epidemiológicos. Os 437 pacientes suspeitos de dengue, de janeiro a junho do presente ano, foram cadastrados no LACEN/SE. Após critérios de exclusão, 382 amostras séricas destes pacientes com diagnóstico e resultado negativo para dengue através da metodologia ELISA NS1 Antígeno do kit Platelia foram analisadas pela técnica de RT-PCR em tempo real no sistema multiplex para a confirmação dos casos suspeitos de dengue. A distribuição dos pacientes de acordo com as variáveis epidemiológicas e demográficas revelou que houve predominância dos casos na região centro-sul do estado e da grande Aracaju (53,5%) podendo-se inferir a facilidade no acesso ao atendimento ambulatorial e laboratorial nessa região, o gênero mais acometido foi o feminino (62,8%) e a faixa etária de maior incidência foi 20 a 59 anos (59,3%). A maior frequência foi na zona urbana (84,9%) e os três primeiros dias dos sintomas (63,9%) com maior índice de coletas. A incidência de dengue nos casos que incialmente se apresentaram como suspeita foi de 22,5% (86) distribuídas entre os sorotipos DENV4 82,5% (71), DENV1 9,3% (8) e DENV3 8,1% (7). Em comparação aos dados oficias que informavam que não houve casos de dengue no referido período, este estudo revela a positividade para dengue com co-circulação de três sorotipos distintos sendo o DENV4 o predominante. Ademias, relata a primeira evidência, dos últimos dez anos, de circulação do sorotipo DENV3 no estado. A análise dos dados permitiu visualizar uma subnotificação dos casos triados e, assim, sugerir uma reavaliação dos métodos utilizados como triagem diagnóstica para que se possam tomar medidas de controle para riscos potenciais de formas graves da doença numa população. / Lagarto, SE

ABSTRACT Introduction Invasive pneumococcal disease (IPD) is an important cause of morbidity and mortality in many parts of the world. It is estimated that pneumococcal disease causes more than one million-childhood deaths every year and the burden of disease is greater in developing countries. The main aim of this study was to analyze risk factors associated with mortality in invasive pneumococcal disease in all ages in South Africa. Materials and Methods We performed an analytical cross-sectional analysis of secondary data from national population-based surveillance for invasive Streptococcus pneumoniae infection in South Africa. The study period was 1 January 2003 to 31 December 2005, and the mortality analysis used a subset of laboratory-confirmed cases who had a completed case report form and available mortality data. Multiple logistic regression models were constructed to identify risk factors significantly associated with the increased risk of death in patients with invasive pneumococcal disease. Separate models were used to evaluate risk factors for death in patients with meningitis and those with other IPD. Results There were 1154 (24%) cases of Streptococcus pneumoniae meningitis and 3736 (76%) cases of other invasive disease. The overall case fatality rate was 1360/4890 (27.8%) of which 911 (67%) patients died within 2 days of admission and 449 (33%) died between 2 days and 30 days of admission. Variables associated with mortality in a logistic regression analysis of all IPD patients included meningitis (OR 2.8, CI 1.9 – 3.9, P=<0.001), HIV-infection (OR 2.8, CI 1.6 – 4.6, P=<0.001), acute severe illness measured by Pitt bacteraemia score >=4 (OR 4.7, CI 2.8 – 7.7, P=<0.001) and prior antibiotic use within 2 months before first positive culture (OR 2.1, CI 1.4 – 3.1, P=<0.001). In addition to this children less than 1 year and adults ≥45 years were more likely to die compared to other age groups. Patients from Western Cape Province were significantly less likely to die (OR 0.27, CI 0.15 – 0.50, P=<0.001) compared to other provinces. Amongst HIV-positive patients severe immunosuppression (low CD4+ count) was a risk factor for death. Risk factors for death were similar in patients with other IPD and meningitis except for HIV which was associated with death in the meningitis group but not in the other IPD group. Antibiotic resistance and vaccine-serotype disease were not associated with increased risk of death. Discussion and Conclusions IPD is associated with a high mortality in South Africa. Our findings of increased risk of death in HIV-positive patients especially those with low CD4+ count are of importance given the high prevalence of HIV amongst patients with IPD. Introduction of the pneumococcal conjugate vaccine as part of the national expanded program for immunization (EPI) and ensuring access to antiretroviral therapy for HIV-positive patients where indicated should be prioritized.

As infecções por Streptococcus pneumoniae (pneumococo) ainda desafiam os sistemas de saúde em todo mundo. Este é um estudo observacional, de seguimento retrospectivo, que avaliou aspectos microbiológicos e clínicos das cepas de pneumococo isoladas de pacientes com doença invasiva pneumocócica (DIP) isolados nos Departamentos Regionais de Saúde (DRS) de Araraquara, Barretos, Franca e Ribeirão Preto, em um período de 16 anos (1998-2013). As informações foram obtidas junto ao Instituto Adolfo Lutz e, no banco de dados do Hospital das Clínicas de Ribeirão Preto (HCRP). Analisou-se 796 linhagens, com predominio do gênero masculino (58,9%), da faixa etária de 20 a menores de 60 anos de idade (32,2%) e do período de 2003 a 2010 (60,2%). As DIPs mais comuns foram a meningite (45,7%) e a pneumonia (45,0%). Quanto aos sorotipos mais frequentes, observou-se em 83,3%: 14, 3, 19F, 1, 6A, 6B, 23F, 9V, 18C, 19A, 12F, 4, 7F, 5, 22F, 11A, 8, 9N, 10A e 15C, sendo o 14 o mais comum nos quatro DRS estudados. Os sorotipos 14, 3 e 19F foram mais frequentes na meningite, enquanto os sorotipos 14, 3 e 1 na pneumonia. Após 2010, verificou-se diminuição dos sorotipos 14, 1, 23F e 5 e aumento de 12F, 11A e 8, não contidos na vacina. A resistência à penicilina foi de 14,8%, sendo 3,0% resistência intermediária e 11,8% de resistência plena. Para ceftriaxona, 5,3% foram não sensíveis. A sensibilidade ao cloranfenicol, eritromicina e ceftriaxona manteve-se acima dos 90%, no período estudado. O maior nível de resistência foi observado para Sulfametoxazol/trimetoprim (49,4%). Destaca-se o aumento dos sorotipos 12F, 11A e 8 após a vacinação, considerando que nenhum deles compõe as vacinas conjugadas disponíveis. Observou-se variabilidade de resistência entre os diferentes sorotipos de pneumococo. A DIP mais frequente nos pacientes cadastrados no HCRP foi a pneumonia (67,8%), seguida da meningite (22,9%) tendo como sorotipos mais frequentes 14, 6A, 23F, 1, 3, 18C, 19F, 12F, 4,9V, 6B e 19A. Destes pacientes 67,5% apresentaram cura sem sequelas, 6,9% tiveram algum tipo de sequela e 25,6% evoluíram para óbito. A pneumonia causou 18,2% dos óbitos, principalmente na faixa etária de 20 a menores de 60 anos de idade. Os sorotipos 12F, 14, 18C, 9V, 18A, 19A e 23F foram responsáveis por 64,9% dos óbitos por meningite, enquanto os sorotipos 3, 14, 9V, 6B, 23F e 19F estiveram envolvidos em 63,4% das mortes por pneumonia. Entre os pacientes que morreram 68,2% tinham algum tipo de comorbidade, sendo HIV/AIDS, alcoolismo e câncer as mais comuns. A faixa etária com 60 anos ou mais foi a mais significativa (OR=4,2) para o insucesso, independente da presença de comorbidade. A presença do sorotipo 18C foi fator de risco significativo tanto na análise bruta (OR=3,8), quanto ao ajustar por comorbidade (OR=5,0) ou ajustada por idade (OR=5,4). O mesmo ocorreu para o sorotipo 12F (respectivamente, OR=5,1, OR=5,0 e OR=4,7). Observou-se alterações na circulação de alguns sorotipos de pneumococo no período pós VPC10. Ressalta-se a importância da continuidade da vigilância das DIPs, afim de determinar oscilações clínicas e microbiológicas da doença. Além disto, na era das vacinas conjugadas, o contínuo monitoramento sobre a distribuição de sorotipos na população é necessário para a avaliação do impacto e adequação da imunização / Infections by Streptococcus pneumoniae (pneumococcus) are still a challenge to health systems worldwide. An observational retrospective study was developed to assess microbiological and clinical aspects of pneumococcus strains isolated from patients with invasive pneumococcal diseases (IPD) which were isolated in the Regional Health Departments (DRS) of Araraquara, Barretos, Franca and Ribeirão Preto, in a period of 16 years (1998-2013). Data were obtained at the Adolfo Lutz Institute and in databases of the Clinics Hospital of Ribeirão Preto (HCRP). A total of 796 strains were analyzed, with prevalence of male individuals (58.9%), aged between 20 and 60 years (32.2%), and in the period between 2003 and 2010 (60.2%). The most common IPD were meningitis (45.7%) and pneumonia (45.0%). Regarding the most frequent serotypes, in 83.3% they were: 14, 3, 19F, 1, 6A, 6B, 23F, 9V, 18C, 19A, 12F, 4, 7F, 5, 22F, 11A, 8, 9N, 10A and 15C, with 14 being the most common in the four DRS studied. Serotypes 14, 3 and 19F were more frequent in meningitis, whereas serotypes 14, 3 and 1 were more frequent in pneumonia. After 2010, there was a decrease in serotypes 14, 1, 23F and 5, and an increase in 12F, 11A and 8, which are not included in the vaccine. Resistance to penicillin was 14.8%, with 3.0% being intermediate, and 11.8% full resistance. For ceftriaxone, 5.3% were not sensitive. Sensitivity to chloramphenicol, erythromycin and ceftriaxone remained over 90% in the studied period. The highest level of resistance was observed for Sulfamethoxazole/trimethoprim (49.4%). It is noteworthy that there was an increase in the s serotypes 12F, 11A and 8 after vaccination, considering that none of them make up the combined vaccines available. Resistance varied among the different serotypes of pneumococcus. The most frequent IPD in the patients registered in the HCRP was pneumonia (67.8%), followed by meningitis (22.9%), with the most frequent serotypes being 14, 6A, 23F, 1, 3, 18C, 19F, 12F, 4, 9V, 6B and 19A. Of these patients, 67.5% were cured without sequela, 6.9% had some sort of sequela and 25.6% evolved to death. Pneumonia caused 18.2% of the deaths, mainly in the age range between 20 and 60 years. Serotypes 12F, 14, 18C, 9V, 18A, 19A and 23F were responsible for 64.9% of the deaths by meningitis, whereas serotypes 3, 14, 9V, 6B, 23F and 19F were involved in 63.4% of the deaths by pneumonia. Among the patients who died, 68.2% had some sort of comorbidity, with HIV/AIDS, alcoholism and cancer being the most common. The age range over 60 years was the most significant (OR=4.2) for failure, regardless of the presence of a comorbidity. The presence of serotype 18C was a significant risk factor both in the gross analysis (OR=3.8), and in the adjustment as for comorbidity (OR=5.0) or age (OR=5.4). This was also true for the serotype 12F (respectively, OR=5.1, OR=5.0 and OR=4.7). There were alterations in the circulation of some pneumococcus serotypes in the period after VPC10. it is emphasized the importance of continued monitoring of DIPs, in order to determine clinical and microbiological fluctuations of the disease. In addition, in the era of combined vaccines, it is necessary to keep monitoring the distribution of serotypes in the population to assess the impact and adequacy of immunization

As infecções por Streptococcus pneumoniae (pneumococo) ainda desafiam os sistemas de saúde em todo mundo. Este é um estudo observacional, de seguimento retrospectivo, que avaliou aspectos microbiológicos e clínicos das cepas de pneumococo isoladas de pacientes com doença invasiva pneumocócica (DIP) isolados nos Departamentos Regionais de Saúde (DRS) de Araraquara, Barretos, Franca e Ribeirão Preto, em um período de 16 anos (1998-2013). As informações foram obtidas junto ao Instituto Adolfo Lutz e, no banco de dados do Hospital das Clínicas de Ribeirão Preto (HCRP). Analisou-se 796 linhagens, com predominio do gênero masculino (58,9%), da faixa etária de 20 a menores de 60 anos de idade (32,2%) e do período de 2003 a 2010 (60,2%). As DIPs mais comuns foram a meningite (45,7%) e a pneumonia (45,0%). Quanto aos sorotipos mais frequentes, observou-se em 83,3%: 14, 3, 19F, 1, 6A, 6B, 23F, 9V, 18C, 19A, 12F, 4, 7F, 5, 22F, 11A, 8, 9N, 10A e 15C, sendo o 14 o mais comum nos quatro DRS estudados. Os sorotipos 14, 3 e 19F foram mais frequentes na meningite, enquanto os sorotipos 14, 3 e 1 na pneumonia. Após 2010, verificou-se diminuição dos sorotipos 14, 1, 23F e 5 e aumento de 12F, 11A e 8, não contidos na vacina. A resistência à penicilina foi de 14,8%, sendo 3,0% resistência intermediária e 11,8% de resistência plena. Para ceftriaxona, 5,3% foram não sensíveis. A sensibilidade ao cloranfenicol, eritromicina e ceftriaxona manteve-se acima dos 90%, no período estudado. O maior nível de resistência foi observado para Sulfametoxazol/trimetoprim (49,4%). Destaca-se o aumento dos sorotipos 12F, 11A e 8 após a vacinação, considerando que nenhum deles compõe as vacinas conjugadas disponíveis. Observou-se variabilidade de resistência entre os diferentes sorotipos de pneumococo. A DIP mais frequente nos pacientes cadastrados no HCRP foi a pneumonia (67,8%), seguida da meningite (22,9%) tendo como sorotipos mais frequentes 14, 6A, 23F, 1, 3, 18C, 19F, 12F, 4,9V, 6B e 19A. Destes pacientes 67,5% apresentaram cura sem sequelas, 6,9% tiveram algum tipo de sequela e 25,6% evoluíram para óbito. A pneumonia causou 18,2% dos óbitos, principalmente na faixa etária de 20 a menores de 60 anos de idade. Os sorotipos 12F, 14, 18C, 9V, 18A, 19A e 23F foram responsáveis por 64,9% dos óbitos por meningite, enquanto os sorotipos 3, 14, 9V, 6B, 23F e 19F estiveram envolvidos em 63,4% das mortes por pneumonia. Entre os pacientes que morreram 68,2% tinham algum tipo de comorbidade, sendo HIV/AIDS, alcoolismo e câncer as mais comuns. A faixa etária com 60 anos ou mais foi a mais significativa (OR=4,2) para o insucesso, independente da presença de comorbidade. A presença do sorotipo 18C foi fator de risco significativo tanto na análise bruta (OR=3,8), quanto ao ajustar por comorbidade (OR=5,0) ou ajustada por idade (OR=5,4). O mesmo ocorreu para o sorotipo 12F (respectivamente, OR=5,1, OR=5,0 e OR=4,7). Observou-se alterações na circulação de alguns sorotipos de pneumococo no período pós VPC10. Ressalta-se a importância da continuidade da vigilância das DIPs, afim de determinar oscilações clínicas e microbiológicas da doença. Além disto, na era das vacinas conjugadas, o contínuo monitoramento sobre a distribuição de sorotipos na população é necessário para a avaliação do impacto e adequação da imunização / Infections by Streptococcus pneumoniae (pneumococcus) are still a challenge to health systems worldwide. An observational retrospective study was developed to assess microbiological and clinical aspects of pneumococcus strains isolated from patients with invasive pneumococcal diseases (IPD) which were isolated in the Regional Health Departments (DRS) of Araraquara, Barretos, Franca and Ribeirão Preto, in a period of 16 years (1998-2013). Data were obtained at the Adolfo Lutz Institute and in databases of the Clinics Hospital of Ribeirão Preto (HCRP). A total of 796 strains were analyzed, with prevalence of male individuals (58.9%), aged between 20 and 60 years (32.2%), and in the period between 2003 and 2010 (60.2%). The most common IPD were meningitis (45.7%) and pneumonia (45.0%). Regarding the most frequent serotypes, in 83.3% they were: 14, 3, 19F, 1, 6A, 6B, 23F, 9V, 18C, 19A, 12F, 4, 7F, 5, 22F, 11A, 8, 9N, 10A and 15C, with 14 being the most common in the four DRS studied. Serotypes 14, 3 and 19F were more frequent in meningitis, whereas serotypes 14, 3 and 1 were more frequent in pneumonia. After 2010, there was a decrease in serotypes 14, 1, 23F and 5, and an increase in 12F, 11A and 8, which are not included in the vaccine. Resistance to penicillin was 14.8%, with 3.0% being intermediate, and 11.8% full resistance. For ceftriaxone, 5.3% were not sensitive. Sensitivity to chloramphenicol, erythromycin and ceftriaxone remained over 90% in the studied period. The highest level of resistance was observed for Sulfamethoxazole/trimethoprim (49.4%). It is noteworthy that there was an increase in the s serotypes 12F, 11A and 8 after vaccination, considering that none of them make up the combined vaccines available. Resistance varied among the different serotypes of pneumococcus. The most frequent IPD in the patients registered in the HCRP was pneumonia (67.8%), followed by meningitis (22.9%), with the most frequent serotypes being 14, 6A, 23F, 1, 3, 18C, 19F, 12F, 4, 9V, 6B and 19A. Of these patients, 67.5% were cured without sequela, 6.9% had some sort of sequela and 25.6% evolved to death. Pneumonia caused 18.2% of the deaths, mainly in the age range between 20 and 60 years. Serotypes 12F, 14, 18C, 9V, 18A, 19A and 23F were responsible for 64.9% of the deaths by meningitis, whereas serotypes 3, 14, 9V, 6B, 23F and 19F were involved in 63.4% of the deaths by pneumonia. Among the patients who died, 68.2% had some sort of comorbidity, with HIV/AIDS, alcoholism and cancer being the most common. The age range over 60 years was the most significant (OR=4.2) for failure, regardless of the presence of a comorbidity. The presence of serotype 18C was a significant risk factor both in the gross analysis (OR=3.8), and in the adjustment as for comorbidity (OR=5.0) or age (OR=5.4). This was also true for the serotype 12F (respectively, OR=5.1, OR=5.0 and OR=4.7). There were alterations in the circulation of some pneumococcus serotypes in the period after VPC10. it is emphasized the importance of continued monitoring of DIPs, in order to determine clinical and microbiological fluctuations of the disease. In addition, in the era of combined vaccines, it is necessary to keep monitoring the distribution of serotypes in the population to assess the impact and adequacy of immunization

Ovinos são considerados reservatórios de Escherichia coli produtora da toxina Shiga (STEC). Este estudo pesquisou amostras de 100 carneiros no Estado de São Paulo. PCR foi empregada para detecção de stx1, stx2, eae, ehx, e intiminas. RFLP-PCR foi realizado para identificação das variantes de stx1/stx2 e fliC. Cinco isolados eae+stx- de sorotipos O128:H2, O145:H2, O153:H7 e O178:H7 apresentaram intiminas b, g e e. Cinqüenta e seis STEC foram obtidos e os genótipos foram stx1cstx2d-O118 (56,1%), stx1c (33,3%), stx2d-O118 (7,6%), e stx1cstx2dOX3a (3%). Os sorotipos mais freqüentes foram ONT:H8, ONT:H-, O75:H-, O174:H8 e O5:H-. Gene fliC codificando para H2, H8, H14, H16, H19, H21 ou H28 foi identificado em 25/33 isolados. ehx foi detectado em três/cinco EPEC Atípicas e em 38 (57,6%) STEC. Sensibilidade aos antimicrobianos foi observada em 77,2% dos isolados STEC. A análise da similaridade genética por PFGE revelou 24 perfis entre 40 isolados STEC e EPEC atípicas. O estudo demonstrou que ovinos saudáveis podem se comportar como reservatórios de STEC e EPEC atípica. / Lambs are reported as carriers of Shiga toxin-producing Escherichia coli (STEC) worldwide. This study surveyed 100 sheep in São Paulo, Brazil. PCR was used for detection of stx1, stx2, eae, ehx, and intimins. RFLP-PCR investigated the stx1/stx2 variants and flagellar antigen (fliC). Five isolates were eae+/stx-, belonging to serotypes O128:H2, O145:H2, O153:H7 and O178:H7, harboring b, g and e intimins. Fifty-six STEC isolates were recovered, and stx genotypes were stx1cstx2d-O118 (56.1%), stx1c (33.3%), stx2d-O118 (7.6%), and stx1cstx2dOX3a (3%). A diversity of STEC serotypes was observed, but most frequent were ONT:H8, ONT:H-, O75:H-, O174:H8 and O5:H-. fliC gene coding for H2, H8, H14, H16, H19, H21 or H28 was identified in 25/33 isolates. ehx gene was detected in three Atypical EPEC and in 38 (57,6%) STEC. Fifty-one (77.2%) STEC were susceptible to antimicrobials tested. Pulsed field gel electrophoresis (PFGE) revealed 24/40 profiles. The study showed that healthy ovine can be carrier of STEC and atypical EPEC in Brazil.

Shiga toxin-producing <i>E. coli</i> (STEC) is an important food- and water-borne pathogen of humans, causing Hemorrhagic Colitis and Haemolytic Uremic Syndrome. Colonization of both cattle and human hosts is mediated through the action of effector molecules secreted via a type III secretion system (T3SS), which forms attaching and effacing lesions (A/E). The necessary effectors which form A/E by manipulation of host signalling and actin nucleation are present on a pathogenicity island called the Locus of Enterocyte Effacement (LEE).<p> It has been reported that vaccination of cattle with Type III-secreted proteins (T3SPs) from STEC O157 resulted in decreased shedding. In order to extend this to non-O157 STEC serotypes, we examined the serological cross-reactivity of T3SPs of serotypes O26:H11, O103:H2, O111:NM and O157:H7. Groups of cattle were vaccinated with T3SPs produced from each of the serotypes and the magnitude and specificity of the responses were measured resulting in limited cross reactivity. Overall, results suggest that vaccination of cattle with T3SPs as a means of reducing the risk of STEC transmission to humans will induce protection that is serotype specific.<p> To pursue the possibility of a cross-protective vaccine, we investigated the protective properties of a chimeric Tir protein against STEC serotypes. Several studies have reported that Tir is highly immunogenic and capable of producing high antibody titers. Potter and colleagues also demonstrated that the vaccination of cattle with ∆tir STEC O157 strain did not protect as well as the wildtype strain. We constructed thirty-mer peptides to the entire STEC O157 Tir protein, as well as to the intimin binding domain of the Tir protein from STEC serotype O26, O103 and O111. Using sera raised against STEC O157 and non-O157 T3SPs, we identified a number of immunogenic peptides containing epitopes unique to a particular serotype. Two different chimeric Tir proteins were constructed containing the STEC O157 Tir protein fused with six STEC non-O157 peptides with or without the Leukotoxin produced by <i>Mannheimia haemolytica</i>. However, the vaccination of mice with the chimeric protein did not protect against challenge with STEC O157 or STEC O111. These results suggest that to achieve cross protection against STEC serotypes using a recombinant protein vaccine, other immunogenic and protective antigens must also be included.<p> In order to identify other immunogenic and cross-protective antigens we cloned and expressed the genes coding for 66 effectors and purified each as histidine-tagged proteins. These included 37 LEE-encoded proteins and 29 non-LEE effectors. The serological response against each protein was measured by Western blot analysis and an enzyme-linked immunosorbent assay (ELISA) using sera from rabbits immunized with T3SPs from four STEC serotypes, experimentally infected cattle and human sera from 6 HUS patients. A total of 20 proteins were recognized by at least one of the STEC T3SP- vaccinated rabbits using Western blots. Sera from experimentally infected cattle and HUS patients were tested using an ELISA against each of the proteins. Tir, EspB, EspD, EspA and NleA were recognized by the majority of the samples tested. Overall, proteins such as Tir, EspB, EspD, NleA and EspA were highly immunogenic for both vaccinated and naturally infected subjects.<p> Based on the above results, two different mixtures of secreted proteins (5 proteins and 9 proteins) were used to vaccinate mice and test the level of shedding following challenge with STEC O157. Overall, the cocktail vaccine containing 9 immunogenic effectors including Tir, EspB, EspD, NleA and EspA was capable of reducing shedding as effectively as the current STEC T3SPs vaccine, Econiche®.

Shiga toxin-producing <i>E. coli</i> (STEC) is an important food- and water-borne pathogen of humans, causing Hemorrhagic Colitis and Haemolytic Uremic Syndrome. Colonization of both cattle and human hosts is mediated through the action of effector molecules secreted via a type III secretion system (T3SS), which forms attaching and effacing lesions (A/E). The necessary effectors which form A/E by manipulation of host signalling and actin nucleation are present on a pathogenicity island called the Locus of Enterocyte Effacement (LEE).<p> It has been reported that vaccination of cattle with Type III-secreted proteins (T3SPs) from STEC O157 resulted in decreased shedding. In order to extend this to non-O157 STEC serotypes, we examined the serological cross-reactivity of T3SPs of serotypes O26:H11, O103:H2, O111:NM and O157:H7. Groups of cattle were vaccinated with T3SPs produced from each of the serotypes and the magnitude and specificity of the responses were measured resulting in limited cross reactivity. Overall, results suggest that vaccination of cattle with T3SPs as a means of reducing the risk of STEC transmission to humans will induce protection that is serotype specific.<p> To pursue the possibility of a cross-protective vaccine, we investigated the protective properties of a chimeric Tir protein against STEC serotypes. Several studies have reported that Tir is highly immunogenic and capable of producing high antibody titers. Potter and colleagues also demonstrated that the vaccination of cattle with ∆tir STEC O157 strain did not protect as well as the wildtype strain. We constructed thirty-mer peptides to the entire STEC O157 Tir protein, as well as to the intimin binding domain of the Tir protein from STEC serotype O26, O103 and O111. Using sera raised against STEC O157 and non-O157 T3SPs, we identified a number of immunogenic peptides containing epitopes unique to a particular serotype. Two different chimeric Tir proteins were constructed containing the STEC O157 Tir protein fused with six STEC non-O157 peptides with or without the Leukotoxin produced by <i>Mannheimia haemolytica</i>. However, the vaccination of mice with the chimeric protein did not protect against challenge with STEC O157 or STEC O111. These results suggest that to achieve cross protection against STEC serotypes using a recombinant protein vaccine, other immunogenic and protective antigens must also be included.<p> In order to identify other immunogenic and cross-protective antigens we cloned and expressed the genes coding for 66 effectors and purified each as histidine-tagged proteins. These included 37 LEE-encoded proteins and 29 non-LEE effectors. The serological response against each protein was measured by Western blot analysis and an enzyme-linked immunosorbent assay (ELISA) using sera from rabbits immunized with T3SPs from four STEC serotypes, experimentally infected cattle and human sera from 6 HUS patients. A total of 20 proteins were recognized by at least one of the STEC T3SP- vaccinated rabbits using Western blots. Sera from experimentally infected cattle and HUS patients were tested using an ELISA against each of the proteins. Tir, EspB, EspD, EspA and NleA were recognized by the majority of the samples tested. Overall, proteins such as Tir, EspB, EspD, NleA and EspA were highly immunogenic for both vaccinated and naturally infected subjects.<p> Based on the above results, two different mixtures of secreted proteins (5 proteins and 9 proteins) were used to vaccinate mice and test the level of shedding following challenge with STEC O157. Overall, the cocktail vaccine containing 9 immunogenic effectors including Tir, EspB, EspD, NleA and EspA was capable of reducing shedding as effectively as the current STEC T3SPs vaccine, Econiche®.