Wilia Marta Elsner Diederichsen de Brito

Epidemiological features of rotavirus infection in Goiânia, Goiás, Brazil, from 1986 to 2000

A total of 2,605 faecal specimens from children up to 10 years old with or without diarrhoea were collected. Samples were obtained from 1986 to 2000 in hospitals, outpatient clinics and day-care centers in Goiânia, Goiás. Two methodologies for viral detection were utilized: a combined enzyme immunoassay for rotavirus and adenovirus and polyacrylamide gel electrophoresis. Results showed 374 (14.4%) faecal specimens positive for Rotavirus A, most of them collected from hospitalized children. A significant detection rate of rotavirus during the period from April to August, dry season in Goiânia, and different frequencies of viral detection throughout the years of study were also observed. Rotavirus was significantly related to hospitalization and to diarrhoeal illness in children up to 24 months old. This study reinforces the importance of rotavirus as a cause of diarrhoea in children and may be important in regards to the implementation of rotavirus vaccination strategies in our country.

Detection of calicivirus from fecal samples from children with acute gastroenteritis in the West Central region of Brazil

The objective of this study was to describe the circulation of caliciviruses in the West Central region of Brazil and its correlation with childrens gender and age, as well as with the year and months of the sample collection. Reverse transcriptase-polymerase chain reaction was performed to detect the human calicivirus genome in 1006 fecal samples that were collected in Goiânia (n = 696) and Brasília (n = 310). Viral RNA was detected in 8.6% of the samples. No significant difference in viral prevalence was found regarding gender, age or year of the sample. However, it was observed that in Goiânia, there is a higher incidence of caliciviruses from September to March. The analysis employing three primer pairs demonstrated that the Ni/E3 or JV12/13 primer pairs, which detect norovirus (NoV), detected 41 positive samples while the 289/290 primer pair, which detects NoV or sapovirus, detected the remaining 46 samples. Calicivirus circulates in the West Central region of Brazil and for better detection of this virus it is important to use more than one primer pair. Also, we conclude that the seasonality presented by this virus is related to higher humidity in the period.

Soroprevalência e fatores de risco para a infecção pelo herpesvírus bovino tipo 1 (BHV-1) no Estado de Goiás, Brasil

The main objectives of this cross-sectional study were to estimate BHV-1 seroprevalence in a population of non-vaccinated cattle in Goias State, Brazil, and to determine potential risk factors related to the seroprevalence. It was conducted from March to September, 2002. Serum samples were collected from 6,932 animals of 892 herds from 232 municipalities in Goias. Sera were tested for antibodies against BHV-1 using the serum neutralization test. Information regarding animals and herds were recorded through a personal interview with the farmer or farmer manager. The data were analyzed using Epi Info 6.04 and Epi Info for Windows 3.01 programs. The seroprevalence was 51.9%. Eight hundred and seventy nine out of 892 herds (98.5%) had at least one seropositive animal, and all (100%) municipalities showed at least one herd/animal positive. Only age influenced the distribution of neutralization antibodies to this virus in animals. None of the exposure variables analyzed was considered as risk factors for the infection with BHV-1 in cattle herds. With these results we conclude that the infection is spread among cattle herds of municipalities in the state of Goias.

Molecular characterization of the NSP4 gene of human group A rotavirus samples from the West Central region of Brazil

Nonstructural protein 4 (NSP4), encoded by group A rotavirus genome segment 10, is a multifunctional protein and the first recognized virus-encoded enterotoxin. The NSP4 gene has been sequenced, and five distinct genetic groups have been described: genotypes A-E. NSP4 genotypes A, B, and C have been detected in humans. In this study, the NSP4-encoding gene of human rotavirus strains of different G and P genotypes collected from children between 1987 and 2003 in three cities of West Central region of Brazil was characterized. NSP4 gene of 153 rotavirus-positive fecal samples was amplified by reverse transcriptase-polymerase chain reaction and then sequenced. For phylogenetic analysis, NSP4 nucleotide sequences of these samples were compared to nucleotide sequences of reference strains available in GenBank. Two distinct NSP4 genotypes could be identified: 141 (92.2%) sequences clustered with NSP4 genotype B, and 12 sequences (7.8%) clustered with NSP4 genotype A. These results reinforce that further investigations are needed to assess the validity of NSP4 as a suitable target for epidemiologic surveillance of rotavirus infections and vaccine development.

Characterization of mixed infections with different strains of bovine rotavirus in an outbreak of diarrhea in dairy herds in Goiás, Brazil

Ten faecal samples of bovine rotavirus from calves less than 30 days old from an outbreak of diarrhea in Hidrolândia, Goias, Brazil were submitted to serological and molecular characterization, using enzyme immunoassay for subgrouping and serotyping, PAGE for determination of electropherotypes and PCR for genome typing. Nine samples belonged to group A/subgroup I rotavirus and one sample was group A / subgroup non-I/non-II. Four samples were characterized as G10P[11] (B223-like), four samples showed a mixture of two rotavirus strains (G6G10 and P[5]P[11]), one sample was characterized as G6P[11] and one sample was characterized only by G serotyping/genotyping, and did not react with any P primer used. Two electropherotypes were detected and both were present in the same animal. This study demonstrates that two different electropherotypes and/or serotypes of bovine rotavirus can circulate in the same outbreak.

Ocorrência de rotavírus e adenovírus em amostras fecais de crianças com gastrenterite, na cidade de Goiânia

In an attempt to detect rotavirus and adenovirus prevalence among other enteropathogens (bacteria and parasites) in diarrhoea, three hundred fecal samples originating from children living in Goiânia city (Goias state, Brazil) were analysed. Rotavirus was found to be the only pathogen in 47 cases, and associated with other infectious agents in 21 cases. 97,0% positive samples of rotavirus showed an electrophoretic pattern characteristic of subgroup II. Adenovirus was found in 7 cases, and associated with other microrganisms in 1 case. Three methods were applied for virological analyses: enzyme immunoassay for rotavirus and adenovirus (EIARA), polyacrylamide gel electrophoresis (PAGE) and immunoelectron microscopy (IEM). The concordance among the three methods was 92.8%, PAGE and EIARA agreed in 95.8%, and IEM and EIARA agreed in 100.0%.

Molecular characterization of VP6-encoding gene of group A human rotavirus samples from central west region of Brazil

Group A rotaviruses are the main cause of acute gastroenteritis in children worldwide. The intermediate capsid protein VP6 encoded by segment 6 of the dsRNA genome is the major structural component of the virus and it is highly antigenic and immunogenic. VP6 is responsible for group and subgroup (SG) specificities, allowing classification of group A rotavirus into SG I, SG II, SG I + II, and SG non‐I‐non‐II. VP6‐encoding gene of 154 group A human rotavirus samples of different G and P genotypes recovered from children in three cities of Central West region of Brazil was amplified by reverse transcription‐polymerase chain reaction followed by sequencing and phylogenetic analysis. Two distinct genetic groups could be recognized: VP6 genogroups I and II. Sequences analysis also revealed that all samples identified as VP6 genogroup I were associated with NSP4 genotype A, whereas samples identified as VP6 genogroup II were associated with NSP4 genotype B. This is the first study in Central West region regarding genetic variability of the VP6 gene. Further molecular surveillance of rotavirus strains is needed to understand better the occurrence of VP6 gene diversity in Brazil and the significance of VP6 for the control and prevention of rotavirus gastroenteritis. J. Med. Virol. 80:2034–2039, 2008.

Ocorrência de infecção por parvovírus suíno e gastrenterite transmissível em suínos, criados de forma extensiva, em Goiás

The serological status of porcine parvovirus (PPV) infection and transmissible gastroenteritis virus (TGEV) infection were determined in swine from extensive raising systems in the state of Goias, Brazil. Ninety-seven serum samples were collected from animals in 12 extensive farms distributed in six cities located nearby Goiânia, GO, and 74 samples were collected from animals in a slaughterhouse in Goiânia, GO. For the PPV-specific antibody detection, the hemaglutination inhibition test (HI) was used; and for TGE antibody detection, the serum neutralization test was performed. Results showed that 25 out of the total 171 (14.4%) analyzed sera were positive for PPV antibodies, and the HI titers varied between 256 to 4,096. None of the 136 serum samples analyzed for TGEV was positive. This is probably the first study that detected PPV and TGEV-specific antibodies in swine herd in the state of Goias. Data suggest that PPV but not TGEV circulated between and among this population of swine in that state.

A conformational epitope mapped in the bovine herpesvirus type 1 envelope glycoprotein B by phage display and the HSV-1 3D structure.

The selected dodecapeptide (1)DRALYGPTVIDH(12) from a phage-displayed peptide library and the crystal structure of the envelope glycoprotein B (Env gB) from Herpes Simplex Virus type 1 (HSV-1) led us to the identification of a new discontinuous epitope on the Bovine herpesvirus type 1 (BoHV-1) Env gB. In silico analysis revealed a short BoHV-1 gB motif ((338)YKRD(341)) within a epitope region, with a high similarity to the motifs shared by the dodecapeptide N-terminal region ((5)YxARD(1)) and HSV-1 Env gB ((326)YARD(329)), in which the (328)Arg residue is described to be a neutralizing antibody target. Besides the characterization of an antibody-binding site of the BoHV-1 Env gB, we have demonstrated that the phage-fused peptide has the potential to be used as a reagent for virus diagnosis by phage-ELISA assay, which discriminated BoHV-1 infected serum samples from negative ones.