SUMMARYSalmonellosis remains an important cause of diarrheal illness in humans in São Paulo State, Brazil. In this study were identified 3554 Salmonella isolates from human infections, during the period 1996-2003. Among 68 different serovars determined, S. Enteritidis was the most frequent one in gastrointestinal and extra-intestinal infections accounting for 67.4% of all isolates. S. Typhimurium and S. enterica subsp. enterica (4,5,12:i:-) were most frequently isolated from children aged < 1-4 year-old, in contrast, people with S. Enteritidis infections were most likely to be 20-50 year-old. In our geographic area the continued laboratorial surveillance of salmonellosis, including serotyping, has showed the trends in Salmonella serovars causing infections in humans throughout the time.

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Background: Bordetella pertussis is the causative agent of pertussis. In Brazil, laboratory diagnosis of pertussis is based on the culture. In 2010, was standardized the Real-Time PCR TaqMan® in routine diagnosis. Objectives: The aim of this study was to evaluate the impact achieved with the introduction of RT-PCR for the routine diagnosis of pertussis and to compare with the results obtained from culture. Patients and Methods: 4,697 samples of nasopharyngeal secretions collected from suspected pertussis cases and/or contacts were analyzed for RT-PCR and culture, from January 2008 until the end of December 2011. Results: According to the results obtained from the samples 6.9% were culture/RT-PCR positive, 14.8% were positive only for RT-PCR and 0.2% only for culture. Negative samples for both techniques was 3,622 (77.1%) and 1.0% were inconclusive for RT-PCR. Conclusions:The implementation of RT-PCR in routine diagnosis resulted in an increase in laboratory confirmation by almost three times. The RT-PCR assay does not intend to replace the culture technique, but to promote an improvement in the diagnosis of pertussis.

Angiostrongylus cantonensis , a rat lungworm, is one of the leading causes of eosinophilic meningitis in humans. Infection in humans occurs by the ingestion of intermediate hosts, undercooked paratenic hosts or contaminated vegetables and fruits by mucus from infected molluscs. This zoonosis is widespread in tropical and subtropical areas of Southeast Asia, it has also been reported in the Pacific Islands as well as in other regions of Americas. In Brazil, human cases of angiostrongyliasis have been reported since 2007 in Southeast, Northeast, and South regions. In January 2011, we collected a batch of 30 Belocaulus willibaldoi slides in a neighborhood of São Paulo city (Parque Fernanda). Six of them were used for identifying species, and the others (24) were used in parasitological tests through digestion in peptic solution and then larvae isolation by the Rugai method. A total of 250 larvae were obtained and they had morphological traits of Angiostrongy lus spp. Later, four Golden hamsters ( Mesocricetus auratus ) were infected with 38 larvae that allowed the recovery of young worms from the brain and lungs of rodents on the 21 st and 30 th day of infection. In this same neighborhood we captured rodents ( Rattus norvegicus ) that, after necropsy led us to recovery of 22 adult worms in the pulmonary arteries (14 males and 8 females) in May 2011. The larvae and worms obtained from natural infection were evaluated by morphological and morphometric parameters, as well as biological behavior patterns and molecular profile. All methodologies identified the parasite as Angiostrongylus cantonensis . In this way, we report for the first time, the natural infection by A. cantonensis in intermediate ( B. willibaldoi ) and definitive ( R. norvegicus ) hosts in a new urban region of Brazil.

BACKGROUND Angiostrongylus cantonensis is the etiological agent of neuroangiostrongyliasis in humans, which is developed in gastropods and vertebrate species, mainly rodents. Human transmission occurs through consumption of molluscs and paratenic hosts infected with L3, and the migration of larvae to the central nervous system causes eosinophilic meningitis. Laboratory diagnosis is based on molecular and immunological tests, using young or adult females as a source of antigens. However, these tests give positive results only after several weeks of symptoms onset and also cross-reactions with others parasite infections may occur.OBJECTIVES The purpose of this work was to study different antigenic preparations of distinct evolutionary phases of A. cantonensis, in order to improve serological techniques for disease immunodiagnosis.METHODS For this purpose, antigenic fractions of different evolutionary forms were evaluated by Dot-enzyme-linked immunosorbent assay (Dot-ELISA) and Western blot using serum samples.FINDINGS All analysed fractions showed reactivity with serum samples from patients with neuroangiostrongyliasis, especially female membrane alkaline (FAM) and female soluble alkaline (FAS) fractions together with female soluble saline (FSS), improving the technique specificity. MAIN CONCLUSIONSThe results point to the possibility of use of raw female antigens in association with alkaline membrane antigens extracted from adult worms to aid in diagnosis and helps initiate neuroangiostrongyliasis surveillance and control actions.

A Reação de Imunofluorescência Indireta (RIFI) foi utilizada para localizar antígenos em estruturas estágio-específicas em cortes parafinados de vermes fêmeas e em tecidos de caramujos do Gênero Biomphalaria infectados com larvas de terceiro estágio de Angiostrongylus cantonensis. Soros de casos confirmados de meningite eosinfílica foram usados para avaliação da reatividade. Soros não reagentes de casos suspeitos; de pacientes com outras parasitoses e de indivíduos sem outras etiologias foram utilizados como controle de reatividade cruzada. Anticorpos da classe IgG foram reativos para antígenos presentes nos dois estágios e, anticorpos IgM somente para o estágio larvário. Nos cortes de vermes, as marcações fluorescentes foram assinaladas nas cutículas e estruturas internas, com acentuada marcação para os conteúdos uterinos. Nos tecidos dos caramujos as larvas foram encontradas exclusivamente no interior dos granulomas, com marcações fluorescentes nas  cutículas das larvas e no interior dos tecidos granulomatosos. O padrão de fluorescência no granuloma sugere a marcação de antígenos excretores/secretores. Reatividade cruzada mais expressiva ocorreu com anticorpos presentes em soros de pacientes com outras parasitoses, com destaque para estrongiloidíase. A RIFI em cortes parafinados abre novas perspectivas para identificação de antígenos e de marcadores sorológicos, que possam ser aplicados no aprimoramento do diagnóstico laboratorial da meningite eosinofílica.

Background: The introduction of pertussis vaccination in the past decades reduced substantially the number of cases; however, despite the high vaccination coverage rates, pertussis is still a major threat to public health all over the world including Brazil. In this study we present an overview of pertussis in Brazil in the last two decades.Methods: Between 1988 and 2009, the National Reference Laboratory for Pertussis, Instituto Adolfo Lutz, São Paulo, Brazil, received a total of 985 presumptively identified Bordetella spp strains, isolated from sporadic cases and/or some pertussis outbreaks. The strains were forwarded by the Regional Laboratories in São Paulo State and by the Central Public Health Laboratories throughout the country. Identification of the species was done by standard methods. Detection of O1 antigen, and the serotyping were done by slide agglutination test using O1, and Fim 2 and Fim 3 antibodies, respectively. Molecular characterization was done by Pulsed Field Gel Electrophoresis (PFGE).Results: All the 985 strains were confirmed as Bordetella pertussis and the most of them belonged to the serotype 1,3. Strains from southeastern states accounted for 67% (660/985), most of them from São Paulo State (646/985; 65.6%). The remaining 325 strains were from south region (147/985, 15%), northeastern states (87/985, 8.8%), north region (48/985, 5%) and central area (43/985, 4.4%). The great majority (81.1%, 799/985) was from children under twelve years of age, and among them 84.4% (675/799) was from infants aged less than six months. Strains from adolescents and adults accounted for 3.2% (32/985) and 8% (78/985), respectively. The age group was unknown in approximately 7.7%. Strains belonging to several PFGE patterns were identified and some were prevalent.Conclusion: Different B. pertussis clones circulate all over the country. These data do not cover all the country, and may represent only a fraction of the actual number of pertussis cases due to the underreporting, despite pertussis is, in Brazil, a reportable disease since 2001. Data on age distribution of pertussis cases may contribute to develop the policy of booster doses of vaccine in adolescents and adults.