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In Brazil, molecular surveillance expanded after Rotarix™ vaccine introduction, alongside G2P[4] dominance. The G2P[6] genotype, despite sharing the same DS-1-like constellation as G2P[4] strains, remains rare. This retrospective study analyzed eight Brazilian G2P[6] strains (2012–2014) through RT-PCR and 11-segments sequencing, followed by phylogenetic analysis. Two distinct groups were identified: 2012–2013 strains (six) carried a DS-1-like backbone with the rare NSP4 E6 genotype, while 2014 strains (two) exhibited the classical DS-1-like constellation with E2. Phylogenetic analysis confirmed the two main clusters: 2012–2013 strains related to classical G2P[4] and uncommon global genotypes, and 2014 strains resembling emerging DS-1-like G1/G3/G8P[8] reassortants. The 2012–2013 strains clustered within G2-VP7 Lineage IVa, while the 2014 strains belonged to Lineage V, reflecting the global distribution of these variants. All VP4 genes were classified within the P[6]-Ia lineage, with phylogenetic analyses suggesting separate introductions from Asia and Africa. The E6 NSP4 gene segment identified in these strains has an undetermined origin and was not previously associated with G2P[6] strains in Brazil. Despite similarities to G2P[4], G2P[6] strains remain rare, with no genomic features explaining their limited spread. Phylogenetic data indicate multiple reassortment events and international viral exchange, highlighting Brazil’s role in RVA diversity. Ongoing full-genome surveillance is crucial to track rare variants and assess their public health relevance.

Pneumonia and diarrhea are the leading causes of death in children under 5 globally, worsened by viral infections. This study investigates viral agents in children ≤ 3 years with respiratory illness and diarrhea in Metropolitan Region of São Paulo, Brazil, during spring 2021. Twenty paired samples (oropharyngeal swab and feces) were tested using in-house qPCR for HBoV and HAdV, RT-qPCR for RVA, EV, PeV-A, and NoV, and a commercial RT-qPCR kit for SARS-CoV-2, Flu A/B, and RSV. HAstV was detected with conventional nested (RT)-PCR. Positive samples were sequenced for molecular characterization and phylogenetic analysis. Seven viruses were identified: HBoV, NoV, HAdV, PeV-A, EV, RSV, and Flu A. HBoV and NoV were detected in 75% of cases, with co-infection in 65% of patients, indicating their involvement in the gastro-respiratory illness. Genotyping of HBoV (HBoV-1), NoV (GII.4_Sydney[P16], GII.2[P16], and GII.4_Sydney[P31]), EV (Coxsackievirus A6), HAdV (species C, type 6), and PeV-A (genotype 1) showed local virus diversity. Phylogenetic analysis indicated no ongoing community outbreak, with distinct clusters observed. The findings highlight the overlap of respiratory and enteric diseases, revealing local viral diversity and high exposure to enteric viruses. This underscores the challenges in differential diagnosis and the need for syndromic surveillance.

Beginning in late 2016 Brazil faced the worst outbreak of Yellow Fever in recent decades, mainly located in southeastern rural regions of the country. In the present study we characterize the Yellow Fever Virus (YFV) associated with this outbreak in São Paulo State, Brazil. Blood or tissues collected from 430 dead monkeys and 1030 pools containing a total of 5,518 mosquitoes were tested for YFV by quantitative RT-PCR, immunohistochemistry (IHC) and indirect immunofluorescence. A total of 67 monkeys were YFV-positive and 3 pools yielded YFV following culture in a C6/36 cell line. Analysis of five nearly full length genomes of YFV from collected samples was consistent with evidence that the virus associated with the São Paulo outbreak originated in Minas Gerais. The phylogenetic analysis also showed that strains involved in the 2016–2017 outbreak in distinct Brazilian states (i.e., Minas Gerais, Rio de Janeiro, Espirito Santo) intermingled in maximum-likelihood and Bayesian trees. Conversely, the strains detected in São Paulo formed a monophyletic cluster, suggesting that they were local-adapted. The finding of YFV by RT-PCR in five Callithrix monkeys who were all YFV-negative by histopathology or immunohistochemistry suggests that this YFV lineage circulating in Sao Paulo is associated with different outcomes in Callithrix when compared to other monkeys.

The porcine origin rotavirus A (RVA) G5 genotype is notable for its unique and sustained human circulation in Brazil, primarily as G5P[8] during the 1980s–2000s. This study aimed to characterize and investigate the full genome of a rare G5P[6] strain detected in 2013 (RVA/Human-wt/BRA/IAL-R406/2013/G5P[6]) to elucidate its evolutionary origin throughout RT-PCR, sequencing, and phylogenetic analysis. Whole-genome assessment revealed an atypical G5-P[6]-I1-R1-C1-M1-A8-N1-T7-E1-H1 constellation. The IAL-R406 VP7 (classified in Lineage I) was closely related to G5 strains that have circulated in both humans and pigs in Brazil for nearly three decades, showing no evidence of recent variant introduction. The VP4 P[6] (assigned as Lineage I) was genetically similar to Paraguayan and Argentinian G4P[6] porcine-like strains, indicating a regional swine reservoir and zoonotic RVA spillover in South America. The remaining nine segments support the animal–human reassortant origin of IAL-R406, showing broad similarity to porcine-like human and porcine strains described worldwide, with additional relationships to bovine (Republic of Korea, USA), feline-like human (Brazil), equine (UK), simian (Caribbean), wild boar/fox (Croatia), and classical human (Japan, USA) strains. In particular, the NSP1-A8 and NSP3-T7 genotypes, extremely rare in humans yet widespread in animals, especially swine, strongly indicate interspecies reassortment, likely resulting from porcine-to-human transmission. Together, these findings reinforce swine as a persistent reservoir for zoonotic RVA infections and highlight the importance of a One Health approach integrating human and animal surveillance to better understand RVA cross-species transmission and evolution.

During 2006-2011, 5035 fecal samples were tested by PCR for human adenovirus (HAdV) and sequenced. HAdV was detected in 198 cases (3.9%), with the highest rate in children ≤ 5 years. Enteric HAdVs were the most prevalent genotypes (78%; 146/187): HAdV-F41 (63.6%; 119/187), HAdV-F40 (12.3%; 23/187), HAdV-A12 (1.6%; 3/187) and HAdV-A31 (0.5%; 1/187). Non-enteric HAdVs were detected in 22% (41/187): HAdV-C1 (8.0%; 15/187), HAdV-C2 (6.9%; 13/187), HAdV-C5 (4.3%; 8/187), HAdV-D8 (1.3%; 2/187), HAdV-B21 (0.5%; 1/187), HAdV-B3 (0.5%; 1/187) and HAdV-C6 (0.5%; 1/187). This 6-year retrospective study points out a high diversity of HAdV types circulating in Brazil and highlights the need to carry out molecular epidemiological studies of HAdV among patients with acute diarrheal infection on a regular basis.

Rotavirus (RVA) G8 is frequently detected in animals, but only occasionally in humans. G8 strains, however, are frequently documented in nations in Africa. Recently, an increase in G8 detection was observed outside Africa. The aims of the study were to monitor G8 infections in the Brazilian human population between 2007 and 2020, undertake the full-genotype characterization of the four G8P[4], six G8P[6] and two G8P[8] RVA strains and conduct phylogenetic analysis in order to understand their genetic diversity and evolution. A total of 12,978 specimens were screened for RVA using ELISA, PAGE, RT-PCR and Sanger sequencing. G8 genotype represented 0.6% (15/2434) of the entirely RVA-positive samples. G8P[4] comprised 33.3% (5/15), G8P[6] 46.7% (7/15) and G8P[8] 20% (3/15). All G8 strains showed a short RNA pattern. All twelve selected G8 strains displayed a DS-1-like genetic backbone. The whole-genotype analysis on a DS-1-like backbone identified four different genotype-linage constellations. According to VP7 analysis, the Brazilian G8P[8] strains with the DS-1-like backbone strains were derived from cattle and clustered with newly DS-1-like G1/G3/G9/G8P[8] strains and G2P[4] strains. Brazilian IAL-R193/2017/G8P[8] belonged to a VP1/R2.XI lineage and were grouped with bovine-like G8P[8] strains with the DS-1-like backbone strains detected in Asia. Otherwise, the Brazilian IAL-R558/2017/G8P[8] possess a “Distinct” VP1/R2 lineage never previously described and grouped apart from any of the DS-1-like reference strains. Collectively, our findings suggest that the Brazilian bovine-like G8P[8] strains with the DS-1-like backbone strains are continuously evolving and likely reassorting with local RVA strains rather than directly relating to imports from Asia. The Brazilian G8P[6]-DS-1-like strains have been reassorted with nearby co-circulating American strains of the same DS-1 genotype constellation. However, phylogenetic analyses revealed that these strains have some genetic origin from Africa. Finally, rather than being African-born, Brazilian G8P[4]-DS-1-like strains were likely imported from Europe. None of the Brazilian G8 strains examined here exhibited signs of recent zoonotic reassortment. G8 strains continued to be found in Brazil according to their intermittent and localized pattern, thus, does not suggest that a potential emergence is taking place in the country. Our research demonstrates the diversity of G8 RVA strains in Brazil and adds to the understanding of G8P[4]/P[6]/P[8] RVA genetic diversity and evolution on a global scale.

Hand-foot-and-mouth disease (HFMD) is a highly contagious viral disease commonly associated to Enteroviruses (EV). During 2018, Brazil faced massive HFMD outbreaks spread across the country. This study aimed to characterize the EV responsible for the HFMD outbreak that occurred in Paraiba State, Brazilian Northeastern region, in 2018, followed by a phylogenetic analysis to detail information on its genetic diversity. A total of 49 serum samples (one from each patient) collected from children ≤ 15 years old, clinically diagnosed with HFMD were tested for EV using conventional RT-PCR and RT-qPCR. EV infection was confirmed in 71.4% (35/49) of samples. The mean and median ages were 1.83 years and one year old, respectively. Twenty-two EV-positive samples were successfully sequenced and classified as EV-A species; 13 samples were also identified with the CV-A6 genotype. The phylogenetic analysis (VP1 region) of three samples revealed that the detected CV-A6 strains belonged to sub-lineage D3. The CV-A6 strains detected here clustered with strains from South America, Europe and West Asia strains that were also involved in HFMD cases during the 2017-2018 seasons, in addition to the previously detected Brazilian CV-A6 strains from 2012 to 2017, suggesting a global co-circulation of a set of different CV-A6 strains introduced in the country at different times. The growing circulation of the emerging CV-A6 associated with HFMD, together with the detection of more severe cases worldwide, suggests the need for a more intense surveillance system of HFMD in Brazil. In addition, this investigation was performed exclusively on serum samples, and the analysis of whole blood samples should be considered and could have shown advantages when employed in the diagnosis of enteroviral HFMD outbreaks.

This study combined conventional epidemiology of human astroviruses. From 2010 to 2016, 232 stool samples from children under 5 years of age were screened using NGS and conventional RT-PCR followed by genetic analysis in order to investigate the genotypic diversity of classical human astrovirus (HAstV) circulating in Tocantins State, Brazil. HAstV was detected in 16 cases (6.9%). Seven specimens (43.7%; 7/16) were positive according RT-PCR and next-generation sequencing (NGS) to investigate the molecular to both NGS and RT-PCR. NGS and RT-PCR individually revealed six (37.5%; 6/16) and three (18.8%; 3/16) additional positive samples, respectively. Sequencing of the HAstV-positive samples revealed HAstV-1a (9/16), HAstV-4c (3/16), and HAstV-5c (4/16) lineages.

Capybara ( Hydrochoerus hydrochaeris ) is the world’s largest rodent species distributed throughout South America. These animals are incredibly tolerant to anthropogenic environments and are occupying large urban centers. Capybaras are known to carry potentially zoonotic agents, including R . rickettsia , Leishmania spp., Leptospira spp., Trypanosoma spp., Salmonella spp., Toxoplasma gondii , and rabies virus. Focusing on the importance of monitoring potential sources of emerging zoonotic viruses and new viral reservoirs, the aim of the present study was to assess the presence of fecal-borne viruses in the feces of capybaras living in urban parks in São Paulo state, Brazil. A total of 337 fecal samples were collected between 2018 and 2020 and screened for the following: (i) Rotavirus group A (RVA) by ELISA; (ii) non-RVA species and Picobirnavirus (PBV) using PAGE; (iii) Human Bocaparvovirus (HBoV), Bufavirus (BuV), Tusavirus (TuV), and Cutavirus (CuV) qPCR; (iv) Human Enterovirus (EV), Norovirus GII (NoV), and Hantavirus by in houses RT-qPCR; (v) SARS-CoV-2 via commercial RT-qPCR kit assay; and (vi) Astrovirus (AstV) and Adenovirus (AdV) using conventional nested (RT)-PCRs. All fecal samples tested were negative for fecal-borne viruses. This study adds further evidence that the fecal-borne viruses is a minor public health issue in Brazilian capybaras, at least during the surveillance period and surveyed areas. Continuous monitoring of sylvatic animals is essential to prevent and control the emergence or re-emergence of newly discovered virus as well as viruses with known zoonotic potential.

In Brazil, molecular surveillance expanded after Rotarix™ vaccine introduction, alongside G2P[4] dominance. The G2P[6] genotype, despite sharing the same DS-1-like constellation as G2P[4] strains, remains rare. This retrospective study analyzed eight Brazilian G2P[6] strains (2012-2014) through RT-PCR and 11-segments sequencing, followed by phylogenetic analysis. Two distinct groups were identified: 2012-2013 strains (six) carried a DS-1-like backbone with the rare NSP4 E6 genotype, while 2014 strains (two) exhibited the classical DS-1-like constellation with E2. Phylogenetic analysis confirmed the two main clusters: 2012-2013 strains related to classical G2P[4] and uncommon global genotypes, and 2014 strains resembling emerging DS-1-like G1/G3/G8P[8] reassortants. The 2012-2013 strains clustered within G2-VP7 Lineage IVa, while the 2014 strains belonged to Lineage V, reflecting the global distribution of these variants. All VP4 genes were classified within the P[6]-Ia lineage, with phylogenetic analyses suggesting separate introductions from Asia and Africa. The E6 NSP4 gene segment identified in these strains has an undetermined origin and was not previously associated with G2P[6] strains in Brazil. Despite similarities to G2P[4], G2P[6] strains remain rare, with no genomic features explaining their limited spread. Phylogenetic data indicate multiple reassortment events and international viral exchange, highlighting Brazil's role in RVA diversity. Ongoing full-genome surveillance is crucial to track rare variants and assess their public health relevance.In Brazil, molecular surveillance expanded after Rotarix™ vaccine introduction, alongside G2P[4] dominance. The G2P[6] genotype, despite sharing the same DS-1-like constellation as G2P[4] strains, remains rare. This retrospective study analyzed eight Brazilian G2P[6] strains (2012-2014) through RT-PCR and 11-segments sequencing, followed by phylogenetic analysis. Two distinct groups were identified: 2012-2013 strains (six) carried a DS-1-like backbone with the rare NSP4 E6 genotype, while 2014 strains (two) exhibited the classical DS-1-like constellation with E2. Phylogenetic analysis confirmed the two main clusters: 2012-2013 strains related to classical G2P[4] and uncommon global genotypes, and 2014 strains resembling emerging DS-1-like G1/G3/G8P[8] reassortants. The 2012-2013 strains clustered within G2-VP7 Lineage IVa, while the 2014 strains belonged to Lineage V, reflecting the global distribution of these variants. All VP4 genes were classified within the P[6]-Ia lineage, with phylogenetic analyses suggesting separate introductions from Asia and Africa. The E6 NSP4 gene segment identified in these strains has an undetermined origin and was not previously associated with G2P[6] strains in Brazil. Despite similarities to G2P[4], G2P[6] strains remain rare, with no genomic features explaining their limited spread. Phylogenetic data indicate multiple reassortment events and international viral exchange, highlighting Brazil's role in RVA diversity. Ongoing full-genome surveillance is crucial to track rare variants and assess their public health relevance.