As phagocytosis is the first line of defense against malaria, we developed a phagocytosis assay with Plasmodium vivax (P. vivax) merozoites that can be applied to evaluate vaccine candidates. Briefly, after leukocyte removal with loosely packed cellulose powder in a syringe, P. vivax trophozoites matured to the merozoite-rich schizont stages in the presence of the E64 protease inhibitor. The Percoll gradient-enriched schizonts were chemically disrupted to release merozoites that were submitted to merozoite opsonin-dependent phagocytosis in two phagocytic lines with human and mouse antibodies against the N- and C-terminus of P. vivax Merozoite Surface Protein-1 (Nterm-PvMSP1 and MSP119). The resulting assay is simple and efficient for use as a routine phagocytic assay for the evaluation of merozoite stage vaccine candidates.

The bacterial pathogen most commonly associated with endemic forms of childhood diarrhoea is Escherichia coli. Studies of epidemiological characteristics of HEp-2 cell-adherent E. coli in diarrhoeal disease are required, particularly in developing countries. The aim of this study was evaluate the presence and significance of adherent Escherichia coli from diarrhoeal disease in children. The prevalence of LA, AA, and DA adherence patterns were determined in HEp-2 cells, the presence of virulence genes and the presence of the O serogroups in samples obtained from 470 children with acute diarrhoea and 407 controls in Porto Velho, Rondônia, Brazil. E. coli isolates were identified by PCR specific for groups of adherent E. coli. Out of 1,156 isolates obtained, 128 (11.0%) were positive for eae genes corresponding to EPEC, however only 38 (29.6%) of these amplified bfpAgene. EAEC were isolated from 164 (14.1%) samples; of those 41(25%), 32 (19%) and 16 (9.7%) amplified eagg, aggA or aafA genes, respectively and aggA was significantly associated with diarrhoea (P = 0.00006). DAEC identified by their adhesion pattern and there were few isolates. In conclusion, EAEC was the main cause of diarrhoea in children, especially when the aggA gene was present, followed by EPEC and with a negligible presence of DAEC.

Bacterial resistance to commonly used antibiotics has been recognized as a significant global health issue. In this study, we carried out the screening of a family of allylic thiocyanates for their action against a diversity of bacteria and fungi with a view to developing new antimicrobial agents. Allylic thiocyanates bearing halogenated aryl groups, which were readily obtained in two steps from the Morita-Baylis-Hillman adducts, showed moderate-to-high activity against selective pathogens, including a methicillin-resistant S. aureus (MRSA) strain. In particular cases, methyl (Z)-3-(2,4-dichlorophenyl)-2-(thiocyanomethyl)-2-propenoate exhibited antimicrobial activity comparable to the reference antibiotic Imipenem.

Méis são descritos possuindo diferentes propriedades, incluindo a antimicrobiana. Muitos estudos têm apresentado essa atividade de méis produzidos por abelhas Apis mellifera, no entanto estudos incluindo atividades de méis de abelhas sem ferrão são escassos. O objetivo deste estudo foi comparar a atividade antimicrobiana de méis de Melipona compressipes, Melipona seminigra e A. mellifera, coletados no Estado do Amazonas, contra Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Chromobacterium violaceum, e Candida albicans. As concentrações inibitórias mínimas foram determinadas usando o método de diluição em ágar, com ágar Muller-Hinton (para bactérias) ou ágar Sabouraud (para a levedura). S. aureus e E. faecalis foram inibidos por todos os méis em concentrações inferiores a 12%, enquanto E. coli e C. violaceum foram inibidos por méis de abelhas sem ferrão em altas concentrações entre 10 e 20%. A. mellifera inibiu E. coli na concentração de 7% e C. violaceum em baixa concentração (0,7%). C. albicans foi inibida apenas em concentrações entre 30 e 40% dos méis. Assim, todas as variedades de mel testadas apresentaram atividade antimicrobiana sobre os patógenos testados, servindo assim como agente antimicrobiano potencial para diversas abordagens terapêuticas.

Honeys are described possessing different properties including antimicrobial. Many studies have presented this activity of honeys produced by Apis mellifera bees, however studies including activities of stingless bees honeys are scarce. The aim of this study was to compare the antimicrobial activity of honeys collected in the Amazonas State from Melipona compressipes, Melipona seminigra and Apis mellifera against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Chromobacterium violaceum, and Candida albicans. Minimum inhibitory concentrations were determined using the agar dilution method with Müller-Hinton agar (for bacteria) or Saboraud agar (for yeast). Staphylococcus aureus and E. faecalis were inhibited by all honeys at concentrations below 12%, while E. coli and C. violaceum were inhibited by stingless bee honeys at concentrations between 10 and 20%. A. mellifera honey inhibited E. coli at a concentration of 7% and Candida violaceum at 0.7%. C. albicans were inhibited only with honey concentrations between 30 and 40%. All examined honey had antimicrobial activity against the tested pathogens, thus serving as potential antimicrobial agents for several therapeutic approaches.

INTRODUÇÃO: Entre 2000 e 2002, 79 amostras positivas para rotavírus foram coletadas de crianças com diarreia na Amazônia ocidental brasileira. MÉTODOS: Para a caracterização molecular dos genótipos G e P foram realizadas as reações de RT-PCR e a análise dos eletroferotipos por eletroforese em gel de poliacrilamida (PAGE). RESULTADOS: 59 amostras foram confirmadas como pertencentes ao rotavírus grupo A. Os genótipos G1P[8], G3P[8] e G4P[8] apresentaram perfis eletroforéticos longos. O genótipo G1P[8] foi encontrado em maior proporção. O eletroferotipo curto ocorreu apenas em genótipos G2. CONCLUSÕES: A proporção dos genótipos de rotavírus observada não foi diferente de outras áreas do Brasil. Este estudo é a primeira genotipagem de rotavírus na Amazônia ocidental brasileira.

INTRODUCTION: During the period from 2000 to 2002, 79 rotavirus-positive stool samples were collected from children presenting diarrhea in the Western Brazilian Amazon. METHODS: Molecular characterization of the G and P genotypes was performed using RT-PCR and electropherotyping analysis by polyacrylamide gel electrophoresis. RESULTS: A total of 59 samples were confirmed as group A rotavirus. A long electrophoretic profile was exhibited by the G1P[8], G3P[8], and G4P[8] genotypes. The G1P[8] genotype was found in greater proportion. The short electropherotype was exhibited only by G2 genotype strains. CONCLUSIONS: The proportion of the rotavirus genotypes observed was not different from that in other areas of Brazil. This study is the first genotyping of rotavirus in the Western Brazilian Amazon.

The incidence of Shigella spp. was assessed in 877 infants from the public hospital in Rondônia (Western Amazon region, Brazil) where Shigella represents the fourth cause of diarrhea. Twenty-five isolates were identified: 18 were Shigella flexneri, three Shigella sonnei, three Shigella boydii and one Shigella dysenteriae. With the exception of S. dysenteriae, all Shigella spp. isolated from children with diarrhea acquired multiple antibiotic resistances. PCR detection of ipa virulence genes and invasion assays of bloody diarrhea and fever (colitis) were compared among 25 patients testing positive for Shigella. The ipaH and ipaBCD genes were detected in almost all isolates and, unsurprisingly, all Shigella isolates associated with colitis were able to invade HeLa cells. This work alerts for multiple antibiotic resistant Shigella in the region and characterizes presence of ipa virulence genes and invasion phenotypesin dysenteric shigellosis.

Trinta por cento das faringotonsilites agudas são de etiologia estreptocócica com potencial de complicações como a glomerulonefrite difusa aguda e febre reumática. Crianças de creches apresentam maior incidência destas infecções. OBJETIVO: Identificar e comparar a prevalência do Streptococcus pyogenes na orofaringe entre crianças que freqüentam creches e crianças não-institucionalizadas, em duas regiões do Brasil. CASUÍSTICA E MÉTODO: Estudo prospectivo com 200 crianças, provenientes da cidade de Porto Velho/RO e São Paulo/SP, em quatro grupos, freqüentadoras ou não de creches. Realizou-se swab de orofaringe e cultura para identificação do Streptococcus pyogenes. RESULTADOS: Prevalência de 8% e 2% entre as crianças de São Paulo que atendem a creches e do grupo controle, respectivamente, apresentando valor estatístico (p=0,02). Prevalência de 24% e 16% nos grupos de Porto Velho/RO que freqüentam creche e controle respectivamente, não caracterizando diferença estatisticamente significante (p=0,18). Observou-se diferença estatisticamente significante entre os grupos creche e controle de São Paulo/SP aos seus correspondentes de Porto Velho/RO (p < 0,01). CONCLUSÃO: Os resultados deste estudo nos permitem sugerir que freqüência em creches representa um fator de risco para a colonização de orofaringe pelo Streptococcus pyogenes, fato observado em populações distintas, porém com significância estatística em apenas uma das duas amostras.

Thirty percent of acute pharyngotonsillitis is caused by Streptococcus pyogenes, which increased the risk of glomerulonephritis and rheumatic fever. Children attending daycare centers have a higher incidence of these infections. AIM: to identify and compare the prevalence of Streptococcus pyogenes in the oropharynx of children who are enrolled and who are not enrolled in daycare centers in different regions of Brazil. MATERIALS AND METHODS: A prospective study of two hundred children from Sao Paulo/SP and Porto Velho/RO. Children from each city were divided into two groups: those attending, and those not attending daycare centers. Swabs of the oropharynx were taken for bacteriological culture and identification. RESULTS: The prevalence of Streptococcus pyogenes in the Sao Paulo groups were 8% and 2% for daycare and control groups, which was statistically significant (p=0.02). The prevalence in children from Porto Velho/RO was 24% and 16% for daycare and control groups, which was statistically significant (p=0.015). Statistical analysis also showed a significant difference between the corresponding groups in the two locations (p<0.01). CONCLUSION: These results show that daycare attendance is a risk factor for oropharyngeal streptococcal colonization; this was seen in different populations, but was statistically significance in only one of the two samples.

One hundred and thirty cases of diarrhea and 43 age-matched controls, 0 to 5 years old, were studied in a pediatric outpatient unit from a poor peri urban area of Porto Velho, Rondônia. Eighty percent of diarrheal cases were observed in the groups under 2 years of age. Rotavirus (19.2%) was the most frequent enteropathogen associated with diarrhea, followed by Shigella flexneri (6.15%) and S. sonnei (1.5%) and Salmonella sp. (6.9%). Four cases of E. coli enterotoxigenic infections (3.1%), E. coli enteropathogenic (EPEC)(2.3%) one case of E. coli enteroinvasive infection (0.8%) and one case of Yersinia enterocolitica (0.8%) were also identified. Mixed infections were frequent, associating rotavirus, EPEC and Salmonella sp. with Entamoeba histolytica and Giardia lamblia.