Human bocavirus detection and quantification in fecal and serum specimens from recipients of allogeneic hematopoietic stem cell transplantation: A longitudinal study.

OBJECTIVES: The aim of this study was to evaluate the occurrence of human bocavirus (HBoV) and to determine viral loads in samples of patients admitted for allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: Fecal and serum samples were collected from 19 patients, during a 24-month period. Samples were screened by quantitative polymerase chain reaction TaqMan assay, with specific probe and primers targeting the NP1 gene of all HBoVs genotypes (HBoV-1 to - 4), and viral loads were determined using serial dilutions of a recombinant plasmid. RESULTS: HBoV DNA was detected in 42.1% (8 of 19) of the patients in at least one type of sample (feces and/or serum) during the study period, with 75% (6 of 8) of the patients being positive in both types of sample. Viral shedding in feces had a median of 26 days (range, 5 to 121) and viremia was detected in 87.5% (7 of 8) of the patients. The HBoV loads in fecal samples were higher than in sera and, in most cases, HBoV was detected earlier in fecal than in sera samples. In six HBoV-positive patients (6 of 8) diarrhea was observed concomitantly to viral detection in fecal samples. CONCLUSIONS: A high frequency and loads of HBoV in allo-HSCT recipients was observed, especially in fecal samples. Positivity in fecal samples was an early predictor of HBoV presence.

Recombinant noroviruses detected in Mid-West region of Brazil in two different periods 2009-2011 and 2014-2015: Atypical breakpoints of recombination and detection of distinct GII.P7-GII.6 lineages.

Noroviruses are an important cause of acute gastroenteritis. The high incidence of norovirus is a reflection of its great genomic and antigenic variability resultant of evolutionary mechanisms, such as recombination. Herein, the main objective of this study was to characterize partially two regions of norovirus genome (RdRp and VP1) from fecal samples, collected in two different time periods (2009-2011 and 2014-2015) in the Mid-West region of Brazil. Twenty samples were sequenced and characterized (GI.P5-GI.5, GII.P16-GII.3, GI.P7-GI.7, GII.Pe-GII.4 and GII.P7-GII.6). Sequences of GII.Pe-GII.4 genotype were also characterized as Sydney 2012 variant. Genotypes GII.P7-GII.6, GII.P16-GII.3 and GII.Pe-GII.4 (16/20-80%) were identified as norovirus recombinants by phylogeny and bioinformatic analyzes. The GII.P7-GII.6 (62.5%) and GII.Pe-GII.4 (25%) genotypes had recombination point's upstream ORF1/2 overlapping region, whereas GII.P16-GII.3 (12.5%) genotype had the recombination point in the overlapping region. Furthermore, the GII.P7-GII.6, from samples collected in 2009-2011 had different recombinant points than the GII.P7-GII.6 from samples obtained in 2014-2015, forming two different clusters in the phylogenetic analysis. Our study brings information on the circulation of recombinant norovirus genotypes in Mid-West of Brazil, including recombinants with atypical recombination breakpoints, and provides evidence for the circulation of different lineages of the same recombinant genotype.

Sapovirus in Rectal and Nasopharyngeal Swab Samples of Children with Symptoms of Acute Gastroenteritis.

The study included 102 hospitalized children 0-72 months of age, with symptoms of acute gastroenteritis. One fecal and one nasopharyngeal swab sample were obtained from each child. Samples were screened for sapovirus and viral loads were determined. Sapovirus was detected in 18.6% of fecal samples and in 36.3% of nasopharyngeal swab samples. High viral loads were detected.

Identification of Human Bocavirus type 4 in a child asymptomatic for respiratory tract infection and acute gastroenteritis - Goiânia, Goiás, Brazil

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.

Phylogenetic analysis of G1P[8] and G12P[8] rotavirus A samples obtained in the pre- and post-vaccine periods, and molecular modeling of VP4 and VP7 proteins.

Reduction in morbimortality rates for acute gastroenteritis (AGE) by Rotavirus A (RVA) has been observed after the introduction of vaccines, however the agent continues to circulate. The present study described the genomic characterization of the 11 dsRNA segments of two RVA samples G1P[8] obtained in the pre- and post-vaccination periods and one of G12P[8] sample (post-vaccine), compared to Rotarix™ vaccine. Analysis by molecular sequencing of the samples showed that the three samples belonged to genogroup I. In addition, the analysis of VP7 gene revealed that the samples G1 (pre-vaccine), G1 (post-vaccine) and G12 were characterized as lineages II, I and III, respectively. Regarding to VP4 and NSP4 gene it was observed that all samples belonged to lineage III, whereas for VP6 gene, the sample of the pre- and post-vaccine belonged to the lineage IV and I, respectively. Considering the VP7 gene, it was observed high nucleotide and amino acid identity for the two G1 samples when compared to Rotarix™ vaccine and lesser identity for the G12 sample. In relation to antigenic epitope of VP7 greater modifications were observed for the G12 sample in the 7-2 epitope that was confirmed by molecular modeling. On the other hand, for VP4, some changes in the 8-1 and 8-3 antigenic epitopes was observed for the three samples. This data could be interpreted as a low selective pressure exerted by vaccination in relation to G1P[8] samples and lesser protection in relation to G12P[8]. Thus, the continuous monitoring of RVA circulating samples remains important.

Identification of Human Bocavirus type 4 in a child asymptomatic for respiratory tract infection and acute gastroenteritis - Goiânia, Goiás, Brazil.

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.

Circulating profile of rotavirus a, g and p genotypes before and after vaccine introduction in the Brazilian mid-west 1986-2015

The purpose of this study was to perform a comparative analysis of Rotavirus A (RVA) G and P genotypes circulating in the Brazilian Mid-West in the period 1986-2015. Seven studies conducted from 1986 to 2009 were included, as well as fecal samples obtained in the period 2014-2015. RVA was screened by ELISA and/or PAGE; genotyping by conventional RT-PCR and/or genomic sequencing. A temporal variation in the predominance of G genotypes mainly G1 and G2 with G9 and G12 emergence was observed. Even with vaccination, RVA continues to circulate in the population, requiring continuous virus monitoring

Detection of antibodies to Oropouche virus in non-human primates in Goiânia City, Goiás.

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.

Phylodynamics of DENV-1 reveals the spatiotemporal co-circulation of two distinct lineages in 2013 and multiple introductions of dengue virus in Goiás, Brazil.

Dengue virus type 1 (DENV-1) was the first serotype introduced in Brazil, during in the 1980s. Since then, this virus has spread in the Brazilian territory, causing several outbreaks. In 2013 the highest number of dengue cases was notified, when compared to the previous years in Brazil, and the state of Goiás reported over 160 thousand cases. In this study, we aimed to present the Phylodynamics of DENV-1 isolates from the state of Goiás, Brazil, during 2013 outbreak, based on the envelope gene (E) sequences. Phylogenetic analysis revealed that Brazilian DENV-1 isolates are grouped together with viruses from genotype V in two distinct lineages (lineage I and lineage II) reflecting co-circulation. Phylogeographic analyses showed that these lineages were introduced in different moments in Goiás, Brazil, using distinct routes, likely originated from the Caribbean. Lineage I was first introduced coming from Rio de Janeiro (2007-2012), followed by the introduction from Argentina (2010-2013). Lineage II was introduced in a single moment from Rio de Janeiro and this clade has existed since 2007-2010. The different viral introduction events demonstrate the viral dispersion process with neighboring regions, which is essential for the maintenance of outbreaks and introduction of new emerging viruses. In conclusion, obtained data reveals the importance of continuous molecular surveillance of this virus in different regions, providing a better understanding of DENV-1 circulation, considering the evolutionary and virus spread patterns.

Molecular characterization of group A rotavirus before and after the introduction of vaccines in Brazil.

INTRODUCTION: 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.

Hepatitis B virus infection among institutionalized mentally ill patients in Brazil.

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.