Genotyping of group A rotavirus samples from Brazilian children by probe hybridization

The G genotyping of 74 group A rotavirus samples was done by RNA-DNA hybridization (dot-blot) using oligonucleotide probes for the VP7 gene region of the human rotavirus serotypes/genotypes 1, 2, 3 and 4. Thirty-one samples could be genotyped by dot-blot showing the following results: G1 = 16, G4 = 6, G3 = 5, and G2 = 4. The data show circulation of genotypes G1-G4 and the predominance of G1. The knowledge of genotypes provides important information concerning rotavirus circulation in Central Brazil.

Effect of actinomycin D on simian rotavirus (SA11) replication in cell culture

Rotaviruses are the major cause of viral diarrhea in humans and animals. Actinomycin D (Act D) is an antibiotic that intercalates DNA and therefore inhibits DNA-dependent transcription. The current study was carried out to assess the influence of Act D on the replication of simian rotavirus (SA11) in cell culture. Virus-infected MA-104 cell cultures were studied in the presence of Act D at concentrations of 1.25 and 2.5 µg/ml. Treatment of rotavirus-infected cells with 2.5 µg/ml Act D 48 h post-infection reduced the cytoplasmic metachromasia after staining with acridine orange by 25%. Viral RNA labeled with ³H-uridine in the presence of the drug was separated by polyacrylamide gel electrophoresis. Viral RNA replication was not affected by Act D, but increased ³H-uridine uptake was demonstrable by infected cells in the presence of the drug. This possibly was due to the inhibition of cellular RNA synthesis by Act D, which thus enhances incorporation of the radionuclide into the viral RNA. Act D reduced the number of infected cells presenting virus-specific fluorescence 48 h post-infection by more than 50%. These data suggest that Act D may have complexed with viral RNA and prevented newly synthesized mRNA from being translated, but may not have prevented early replication.

Prevalence of diarrheogenic Escherichia coli and rotavirus among children from Botucatu, São Paulo State, Brazil

In a one-year prospective study carried out to define the role of rotavirus and Escherichia coli in local childhood diarrhea, we determined the prevalence of both agents in 54 diarrheic children attending a health center in Botucatu. Diarrheogenic E. coli (DEC) strains were characterized by O:H serotyping, a search for virulence genetic markers, and assays of adherence to HEp-2 cells. Except for enteroaggregative E. coli (EAEC), no other DEC category was detected in the children's stools. Both EAEC and rotavirus were isolated from 22 of the 54 (41.0%) diarrheic children as single agents or in combination with other enteropathogens. However, when considering the presence of a single agent, EAEC was dominant and isolated from 20.4% of the patients, whereas rotavirus was detected in 14.8%. These results indicate that rotavirus and EAEC play a significant role as agents of childhood diarrhea in the local population.

Genetic analysis of the porcine group B rotavirus NSP2 gene from wild-type Brazilian strains

Group B rotaviruses (RV-B) were first identified in piglet feces, being later associated with diarrhea in humans, cattle, lambs, and rats. In human beings, the virus was only described in China, India, and Bangladesh, especially infecting adults. Only a few studies concerning molecular analysis of the RV-B NSP2 gene have been conducted, and porcine RV-B has not been characterized. In the present study, three porcine wild-type RV-B strains from piglet stool samples collected from Brazilian pig herds were used for analysis. PAGE results were inconclusive for those samples, but specific amplicons of the RV-B NSP2 gene (segment 8) were obtained in a semi-nested PCR assay. The three porcine RV-B strains showed the highest nucleotide identity with the human WH1 strain and the alignments with other published sequences resulted in three groups of strains divided according to host species. The group of human strains showed 92.4 to 99.7% nucleotide identity while the porcine strains of the Brazilian RV-B group showed 90.4 to 91.8% identity to each other. The identity of the Brazilian porcine RV-B strains with outer sequences consisting of group A and C rotaviruses was only 35.3 to 38.8%. A dendrogram was also constructed to group the strains into clusters according to host species: human, rat, and a distinct third cluster consisting exclusively of the Brazilian porcine RV-B strains. This is the first study of the porcine RV-B NSP2 gene that contributes to the partial characterization of this virus and demonstrates the relationship among RV-B strains from different host species.

Changes in epidemiology of rotavirus in the Triângulo Mineiro region of Brazil: lack of two consecutive rotavirus seasons

Rotaviruses are the main cause of infantile acute diarrhea, and a monovalent (G1P[8]) vaccine against the virus was introduced into the Brazilian National Immunization Program for all infants in March 2006. The objectives of this study were to determine the rate and genotype distribution of rotavirus causing infantile diarrhea in the Triângulo Mineiro region of Brazil during 2011-2012 and to assess the impact of local vaccination. Fecal specimens were analyzed for detection and characterization of rotavirus using polyacrylamide gel electrophoresis, reverse transcription followed by polymerase chain reaction (PCR), and PCR-genotyping assays. Overall, rotavirus was diagnosed in 1.7% (6/348) of cases. Rotavirus positivity rates decreased 88% [95% confidence intervals (CI)=15.2, 98.3%; P=0.026] in 2011 and 78% (95%CI=30.6, 93.0%; P=0.007) in 2012 when compared with available data for baseline years (2005/2006) in Uberaba. In Uberlândia, reductions of 95.3% (95%CI=66.0, 99.4%; P=0.002) in 2011, and 94.2% (95%CI=56.4, 99.2%; P=0.004) in 2012 were also observed compared with data for 2008. The circulation of rotavirus G2P[4] strains decreased during the period under study, and strains related to the P[8] genotype reemerged in the region. This study showed a marked and sustained reduction of rotavirus-related cases, with a lack of rotavirus in the 2011 and 2012 seasons, suggesting a positive impact of the vaccination program.

Indícios de Rotavirus na etiologia de um surto de infecção de origem alimentar

Doenças transmitidas por alimentos têm merecido especial atenção por parte de pesquisadores em todo mundo. Os surtos alimentares de etiologia viral têm sido tratados inadequadamente como sendo de menor importância quando comparados com aqueles de etiologia bacteriana. O rotavírus é um importante agente causador de doenças diarréicas graves, sendo responsável por significativo número de óbito em vários países. Este trabalho é um relato de surto de gastroenterite ocasionado por rotavírus, ocorrido em estabelecimento comercial com 720 funcionários, dos quais 51 relataram sintomas de mal-estar, vômito, diarréia, náusea e calafrios. Foi realizada análise microbiológica da água, coletada em diversos pontos, bem como de todos os alimentos e bebidas oferecidos para consumo dos funcionários. Nenhum destes apresentou contaminação por microbiota bacteriana patogênica, nem alterações em suas características sensoriais e organolépticas. De acordo com a estimativa do risco atribuível a cada um dos alimentos, o pão com manteiga oferecido foi apontado como alimento suspeito, somado ao fato de que o único manipulador incumbido do preparo deste produto apresentava-se com infecção por rotavírus.

Variabilidad de electroferotipos y serotipos G y P de rotavirus en casos de gastroenteritis infantil [por] Juan Fco. Contreras Cordero

Tesis (Maestría en Ciencias con Especialidad en Inmunobiología) U.A.N.L.

Determinación de la especificidad de la respuesta inmune humoral en una infección natural por rotavirus [por] Griselda Edith Menchaca Rodríguez

Tesis (Maestría en Ciencias con Especialidad en Microbiología) U.A.N.L.

Análisis del efecto antiviral de Waltheria americana L (Standley y Steyermarl, 1949) en el proceso infectivo de rotavirus humano sobre células MA 104.

Tesis (Maestría en Ciencias con acentuación en Microbiología) UANL, 2014.

Bases moleculares de la respuesta inmune en una infección natural por rotavirus

Tesis (Doctorado en Ciencias con Especialidad en Microbiología) UANL

Epitopes inmunodominantes en la subunidad VP8* de la proteínaVP4 de cepas humanas de rotavirus con genotipo P [8].

Tesis (Doctor en Ciencias con Especialidad en Microbiología) UANL, 2013.

Proteína NSP4 de rotavirus como factor de patogénesis.

Tesis (Doctor en Ciencias con Especialidad en Microbiología) UANL, 2013.

Efectividad de la vacuna contra rotavirus en la hospitalización por diarrea en niños menores de dos años en cinco ciudades de Colombia

Introducción: La enfermedad diarreica aguda (EDA) es la segunda causa de muerte a nivel mundial en menores de cinco años. El rotavirus es el principal entero-patógeno asociado a EDA severa por lo cual se ha iniciado la vacunación contra este virus en Colombia. Este trabajo evaluó la efectividad de la vacunación contra la hospitalización por diarrea en niños menores de dos años en cinco ciudades de Colombia. Metodología: Análisis de sobrevida para estimar la efectividad de la vacuna contra el rotavirus en la prevención de hospitalización por EDA, basado en los datos de una encuesta poblacional realizada en niños menores de dos años de Bogotá, Barranquilla, Cali, Cartagena y Riohacha, en la que se recolectó información socio-demográfica, de vacunación, factores de riesgo para EDA y antecedentes de EDA. Resultados: Incidencia acumulada de hospitalización por diarrea de 5,3%. Cobertura de vacunación fue de 88,5% y 78% para primera y segunda dosis respectivamente. La vacunación con dos dosis presentó una efectividad de 52,4% IC 95% (13,2% a 73,9%) en la prevención de hospitalizaciones por EDA y la vacunación adecuada una efectividad de 59,5% IC 95% (23,9% a78,4%). El sexo masculino y la ciudad de residencia (Riohacha) se asociaron al riesgo de hospitalización. Conclusiones: La vacunación contra rotavirus mostro ser efectiva en la prevención de la hospitalización por EDA. Las coberturas de vacunación y la adherencia al esquema presentan deficiencias que requieren medidas para asegurar la vacunación de todos los niños antes de los seis meses..

Economic Impact of a Rotavirus Vaccine in Brazil

The study was done to evaluate the cost-effectiveness of a national rotavirus vaccination programme in Brazilian children from the healthcare system perspective. A hypothetical annual birth-cohort was followed for a five-year period. Published and national administrative data were incorporated into a model to quantify the consequences of vaccination versus no vaccination. Main outcome measures included the reduction in disease burden, lives saved, and disability-adjusted life-years (DALYs) averted. A rotavirus vaccination programme in Brazil would prevent an estimated 1,804 deaths associated with gastroenteritis due to rotavirus, 91,127 hospitalizations, and 550,198 outpatient visits. Vaccination is likely to reduce 76% of the overall healthcare burden of rotavirus-associated gastroenteritis in Brazil. At a vaccine price of US$ 7-8 per dose, the cost-effectiveness ratio would be US$ 643 per DALY averted. Rotavirus vaccination can reduce the burden of gastroenteritis due to rotavirus at a reasonable cost-effectiveness ratio.

Rotavirus Gastroenteritis in Children in 4 Regions in Brazil: A Hospital-Based Surveillance Study

Background. Rotavirus is a major cause of gastroenteritis in children. Knowledge of rotavirus genotypes is important for vaccination strategies. Methods. During 2005-2006, rotavirus surveillance studies were conducted in Sao Paulo, Salvador, Goiania, and Porto Alegre, Brazil. Stool samples were collected from children <5 years of age who had diarrhea and were screened by the Rotaclone Enzyme Immunoassay for the presence of rotavirus. Confirmed rotavirus-positive samples were characterized for P and G genotypes by reverse-transcriptase polymerase chain reaction. Results. A total of 510 stool samples were collected. Of these, 221 (43.3%) were positive for rotavirus. Overall, G9 was the predominant G type, followed by G2, and G1; P[4] and P[8] were the predominant P types. The most frequent G/P genotype combination detected was G2P[4], followed by G9P[8], G9P[4], and G1P[8]. G2P[4] was the predominant type in Goiania and Salvador; G9P[8] and G1P[8] were predominant in Sao Paulo and Porto Alegre, respectively. Conclusions. The prevalence, seasonality, and genotype distribution of rotavirus infection varied in different regions in Brazil. With immunization programs, continuous monitoring of rotavirus types is important to detect novel and emerging strains.