Abstract Human Bocavirus (HBoV) has been identified from feces and respiratory samples from cases of both acute gastroenteritis and respiratory illness as well as in asymptomatic individuals. The aim of this study was to detect and characterize HBoV from fecal samples collected from hospitalized children aged less than five years old with no symptoms of respiratory tract infection (RTI) or acute gastroenteritis (AGE). The study involved 119 children and one fecal sample was collected from each participant between 2014 and 2015. HBoV was detected using Nested-PCR, and the viral type identified by genomic sequencing. HBoV-4 was identified from one sample obtained from a hospitalized child with soft tissue tumor of the submandibular region. This is the first report of HBoV-4 identification in Brazil, but we consider that this type may be circulating in the country similar to the other types and new investigations are necessary.

Abstract: INTRODUCTION Arboviruses are associated with human disease, and non-human primates (NHPs) are important primary hosts. This study shows the detection of antibodies to Oropouche virus (OROV) in NHPs either living in urban parks or acclimatized at the Wild Animal Screening Center, Goiânia city. METHODS: Fifty blood samples were analyzed by hemagglutination-inhibition and neutralization assays. RESULTS: Two monkeys (Alouatta caraya) had antibodies to OROV by both techniques. CONCLUSIONS This is the first report demonstrating the detection of OROV antibodies in Goiás State and may represent the introduction/circulation of OROV in the region and a potential risk to the human population.

ABSTRACT OBJECTIVES: The main objective was to evaluate HBV infection and occult HBV infection (OBI) cases in mentally ill patients based on serological and molecular profiles. MATERIAL AND METHODS: Serum samples of 333 long-stay mentally ill patients were tested for the prevalence of HBV markers by serological (ELISA) and molecular (PCR) assays. The PCR products were sequenced to determine viral genotypes. RESULTS: It was observed a global prevalence of 12.9% (43/333) for HBV infection markers, considering HBsAg and/or anti-HBc positivity. Fourteen samples tested positive for anti-HBs alone. All samples positive (n= 57) for any HBV serological markers were tested for HBV-DNA and six were positive: HBsAg/anti-HBc (n = 1), anti-HBc/anti-HBs (n = 1), anti-HBs alone (n = 1), and anti-HBc alone (n = 3). The rate of OBI was 9.2% (5/54) from samples that were anti-HBc and/or anti-HBs positive. All sequenced samples were characterized as genotype A. CONCLUSION: The high rate of HBV infections found in this study suggests the possibility of HBV transmission due to risk factors displayed by some patients, and highlights the importance of vaccination of susceptible patients and the staff of that institution.

ABSTRACTINTRODUCTION:In this study, the molecular characteristics of group A rotavirus (RVA) were compared in samples obtained before and after RVA vaccine-introduction in Brazil.METHODS:Eighty samples were screened for the presence of RVA. Positive samples were molecularly analyzed.RESULTS:RVA positivity was 16.9%, with a predominance of G2P[4]. Periods: pre-vaccination: predominance of IId (G1), IId (G2) lineages, and I1 and E1 genotypes; post-vaccination: predominance of Ib (G1), IIa, and IIc (G2) lineages and I2 and E2 genotypes.CONCLUSIONS:Although changes in RVA-circulation pattern were observed in the post-vaccination period, it could not be attributed to vaccination process.

To determine the positivity rate of human bocavirus (HBoV) 1 and 3 among children who presented with acute gastroenteritis symptoms during the period of 1994-2004 in the Central-West Region of Brazil, 762 faecal samples were tested using polymerase chain reaction (PCR) for the detection of HBoV DNA. Primers for a segment of the non-structural viral protein 1 (NS1) gene of HBoV-1 and HBoV-3 were used. Twelve HBoV-positive samples were further characterised via genomic sequencing and phylogenetic analysis. Of the samples tested, 5.8% (n = 44) were positive for HBoV-1 or HBoV-3 and co-infection was observed in 14 (31.8%) of the 44 HBoV-positive samples. Nine of the 14 samples were also positive for Rotavirus A and five were positive for Aichi virus. The genomic sequencing of the NS1 partial sequence of 12 HBoV-samples showed that 11 samples were characterised as HBoV-1 and that one was characterised as HBoV-3. The phylogenetic analysis showed that the HBoV-1 samples had a high sequence homology to others previously identified in China, Sweden and Brazil. This is the first study conducted in the Central-West Region of Brazil to detect HBoV-1 and HBoV-3 in faecal samples from children with acute gastroenteritis. Further studies are required to define the role of HBoVs as aetiological agents of gastroenteritis.

The epidemiological features of rotavirus A (RVA) infection differ between children from developing and developed countries which could result in differences in vaccine efficacy around the world. To evaluate the impact of RotarixTM on RVA prevalence, we monitored RVA genotypes circulating in Goiânia by monitoring virus in faecal samples from children that had or had not been previously vaccinated. From February-November of 2008, 220 faecal samples were collected from children in seven day-care centres. RVA detection was performed by two methodologies and the results were confirmed by polyacrylamide gel electrophoresis. From the 220 samples, eight were RVA-positive (3.6%) and five were from children that had received either one or two doses of the vaccine. All positive samples were collected from children with diarrhoea during August and September. Genotyping of the RVA characterised five of the viral samples as genotype G2P[4] and one as G8P[4], suggesting that G2P[4] was the predominant circulating genotype in Goiânia during the study. The fact that vaccinated children were also infected by RVA suggests that the vaccine does not fully protect against infection by the G2[P4] RVA genotype.

Hepatitis A virus (HAV) infection is a public health problem worldwide and the virus has been classified into six genotypes. In Brazil, the only genotype that has been found is genotype I, predominately from subgenotype IA. Here, the HAV genotypes were analyzed of 18 isolates circulating between 1996-2001 in Goiânia, state of Goiás, Brazil. Viral RNA was extracted from 18 serum samples and amplified (RT-PCR/nested-PCR), followed by the genomic sequencing of the VP1/2A junction region of the HAV genome. Sequences of 168 nucleotides were compared and analyzed using the BLAST N, Clustal X and PAUP v. 4.10b programs. All samples were classified as genotype I, with 10 belonging to subgenotype IA and eight to subgenotype IB. The subgenotype IA isolates showed greater diversity than the subgenotype IB isolates at the nucleotide level. Elevated identity values were found between isolates obtained in this study and those from other regions of the world, including Brazil, highlighting the high conservation among different isolates of this virus. However, changes in the HAV subgenotype circulation could also be observed during the evaluated period.

We analyzed fecal samples from hospitalized children up to three years of age with acute gastroenteritis at Campo Grande, Mato Grosso do Sul, Brazil, from May 2000-January 2004. Astrovirus and calicivirus were detected by Reverse Transcription-Polymerase Chain Reaction and adenovirus was detected using the Rotavirus and Adenovirus combined immunoenzyme assay. Astrovirus, adenovirus and calicivirus were detected at rates of 3.1%, 3.6% and 7.6%, respectively. These results re-emphasize the need for the establishment of regional vigilance systems to evaluate the impact of enteric viruses on viral gastroenteritis.

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.

Através da eletroforese em gel de poliacrilamida e do ensaio imunenzimático combinado para rotavírus e adenovirus, foram analisadas 380 amostras fecais de crianças com até 3 anos, hospitalizadas com diarréia aguda, entre maio de 2000 e janeiro de 2004, em Campo Grande, MS. Do total de amostras, 88 (23,2%) foram positivas para Rotavirus A. Dentre essas, 81 (92%) tiveram padrão eletroferotípico definido, sendo 77 (87,5%) de padrão longo e quatro (4,5%) de padrão curto. A caracterização genotípica G e P foi feita por RT-Nested-PCR para 85 amostras, sendo 56 (65,9%) genotipáveis para genótipo G. Dentre essas, 49 (87,5%) foram G1, cinco (8,9%) G4, uma (1,8%) G3 e uma (1,8%) G9. Considerando a genotipagem P, 37 (43,5%) foram genotipáveis e todas eram P[8]. A associação G e P mais observada foi G1P[8], 33 (89,2%) amostras; seguida de G4P[8], duas (5,4%) amostras; G3P[8], uma (2,7%) amostra; e G9P[8], uma (2,7%) amostra.

Polyacrylamide gel electrophoresis and combined immunoenzyme assay for rotavirus and adenovirus were used to analyze 380 fecal samples from children up to three years of age who were hospitalized with acute diarrhea in Campo Grande, State of Mato Grosso do Sul, between May 2000 and January 2004. Among all the samples, 88 (23. 2%) were positive for Rotavirus A. Out of these, 81 (92%) had a defined electrophoretic pattern: 77 (87. 5%) with a long pattern and four (4. 5%) with a short pattern. Genotype G and P characterization was done by nested RT-PCR for 85 samples, of which 56 (65. 9%) were genotyped as type G. Among these, 49 (87. 5%) were G1, five (8. 9%) were G4, one (1. 8%) was G3 and one (1. 8%) was G9. The genotype was found to be type P in 37 samples (43. 5%) and all of these were P[8]. The G and P association most observed was G1P[8], with 33 samples (89. 2%), followed by G4P[8], two samples (5. 4%); G3P[8], one sample (2. 7%); and G9P[8], one sample (2. 7%).

The objective of this study was to describe the circulation of caliciviruses in the West Central region of Brazil and its correlation with children's 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.

In this study, a total of 865 serum samples were collected between 1995 and 2002 from individuals living in Goiânia, Central Brazil, and clinically suspected of hepatitis. After exclusion of 162 samples which were positive for hepatitis B virus or hepatitis C virus, 703 samples were tested for anti-hepatitis A virus (anti-HAV) IgM antibodies by enzyme immunoassay. In addition, 588 of these samples and 22 fecal samples were analyzed by reverse transcription-nested PCR for HAV RNA detection, with positivity indices of 13.1% (77/588) and 54.5% (12/22), respectively. A similar index of viral RNA detection in anti-HAV-IgM positive or negative samples was observed in serum samples. HAV infection is a public health problem worldwide and this study underscores the extent of HAV circulation in our region.

Neste estudo, foram analisadas 648 amostras de soro, provenientes de profissionais de laboratório de Goiânia-Goiás, visando a detecção de três marcadores sorológicos do VHB: HBsAg, anti-HBsAg e anti-HBcAg. As amostras HBsAg e anti-HBcAg positivas foram também analisadas para os marcadores HBeAg, anti-HBeAg e anti-HBcAgIgM. Foi observado um percentual de soropositividade para infecção pelo VHB de 24,1% sendo que 0,7% destes foram positivos para o HBsAg. O DNA viral foi detectado por PCR nas duas amostras HBsAg positivas. Dos 648 profissionais, 74,5% relataram vacinação para o VHB e, destes, 89,9% apresentaram soroconversão. Trabalho direto com fluidos biológicos bem como trabalho em serviços gerais representaram risco significativo para aquisição da infecção pelo VHB. Os resultados do estudo mostram que embora tenha havido um aumento no índice de vacinação entre os trabalhadores de laboratório, o índice de infecção pelo VHB manteve-se inalterado em relação ao tempo na região.

Were analyzed 648 serum samples from laboratory staff in Goiânia, Goiás aiming detection of three serological markers of HBV: HBsAg, anti-HBsAg and anti-HBcAg. The HBsAg and anti-HBcAg positive samples were also analyzed for HBeAg, anti-HBeAg and anti-HBcAgIgM markers. HBV infection rate of 24.1% was observed and, from them, 0.7% were positive for HBsAg. Viral DNA was detected by PCR in two HBsAg positive samples. A vaccination index of 74.5% and a global index of 89.9% of serological response to vaccination were observed. The direct work with biological fluids as well as cleaning workers represented significant risks for acquisition of HBV infection. The data from the present study showed an increase of the vaccination index among laboratory staff but the rates of HBV infection did not change through the years in the region.