Human bocavirus (HBoV) is an emerging virus and has been detected worldwide, especially in pediatric patients with respiratory and gastrointestinal infection. In this study, we describe HBoV prevalence, genotypes circulation and DNA shedding, in stool samples from children up to two years of age in Brazil. During 2016 and 2017, 886 acute gastroenteritis (AGE) stool samples from ten Brazilian states were analyzed by TaqMan®-based qPCR, to detect and quantify HBoV. Positive samples were genotyped by sequencing the VP1/2 overlap region, followed by phylogenetic analysis and co-infections were accessed by screening other gastroenteric viruses. HBoV was detected in 12.4% (n = 110) of samples, with viral load ranging from 1.6 × 102 to 1.2 × 109 genome copies per gram of stool. From these, co-infections were found in 79.1%, and a statistically lower HBoV viral load was found compared to viral loads of rotavirus, norovirus and adenovirus in double infected patients (p < 0.05). No significant differences were found between HBoV viral load in single or co-infections, age groups or genotypes. Phylogenetic analysis identified the circulation of HBoV-1 in 38%, HBoV-2 in 40% and HBoV-3 in 22%. Continuous HBoV monitoring is needed to clarify its role in diarrhea disease, especially in the absence of classic gastroenteric viruses.

Rotavirus A (RVA) vaccines succeeded in lowering the burden of acute gastroenteritis (AGE) worldwide, especially preventing severe disease and mortality. In 2019, Brazil completed 13 years of RVA vaccine implementation (Rotarix™) within the National Immunization Program (NIP), and as reported elsewhere, the use of Rotarix™ in the country has reduced childhood mortality and morbidity due to AGE. Even though both marketed vaccines are widely distributed, the surveillance of RVA causing AGE and the monitoring of circulating genotypes are important tools to keep tracking the epidemiological scenario and vaccines impact. Thus, our study investigated RVA epidemiological features, viral load and G and P genotypes circulation in children and adults presenting AGE symptoms in eleven states from three out of five regions in Brazil. By using TaqMan®-based one-step RT-qPCR, we investigated a total of 1536 stool samples collected from symptomatic inpatients, emergency department visits and outpatients from January 2018 to December 2019. G and P genotypes of RVA-positive samples were genetically characterized by multiplex RT-PCR or by nearly complete fragment sequencing. We detected RVA in 12% of samples, 10.5% in 2018 and 13.7% in 2019. A marked winter/spring seasonality was observed, especially in Southern Brazil. The most affected age group was children aged >24–60 months, with a positivity rate of 18.8% (p < 0.05). Evaluating shedding, we found a statistically lower RVA viral load in stool samples collected from children aged up to six months compared to the other age groups (p < 0.05). The genotype G3P[8] was the most prevalent during the two years (83.7% in 2018 and 65.5% in 2019), and nucleotide sequencing of some strains demonstrated that they belonged to the emergent equine-like G3P[8] genotype. The dominance of an emergent genotype causing AGE reinforces the need for continuous epidemiological surveillance to assess the impact of mass RVA immunization as well as to monitor the emergence of novel genotypes.

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