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The introduction of rotavirus A (RVA) vaccines has considerably reduced the RVA-associated mortality among children under 5 years of age worldwide. The ability of RVA to reassort gives rise to different combinations of surface proteins G (glycoprotein, VP7) and P (protease sensitive, VP4) RVA types infecting children. During the epidemiological surveillance of RVA in the Northwest Amazon region, an unusual rotavirus genotype G6P[8] was detected in feces of a 2-year-old child with acute gastroenteritis (AGE) that had been vaccinated with one dose of Rotarix® (RV1). The G6P[8] sample had a DS-1-like constellation with a Wa-like VP3 gene mono-reassortment similar to equine-like G3P[8] that has been frequently detected in Brazil previously. The results presented here reinforce the evolutionary dynamics of RVA and the importance of constant molecular surveillance.
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Sapovirus is an important etiological agent of acute gastroenteritis (AGE), mainly in children under 5 years old living in lower-income communities. Eighteen identified sapovirus genotypes have been observed to infect humans. The aim of this study was to identify sapovirus genotypes circulating in the Amazon region. Twenty-eight samples were successfully genotyped using partial sequencing of the capsid gene. The genotypes identified were GI.1 (n = 3), GI.2 (n = 7), GII.1 (n = 1), GII.2 (n = 1), GII.3 (n = 5), GII.5 (n = 1), and GIV.1 (n = 10). The GIV genotype was the most detected genotype (35.7%, 10/28). The phylogenetic analysis identified sapovirus genotypes that had no similarity with other strains reported from Brazil, indicating that these genotypes may have entered the Amazon region via intense tourism in the Amazon rainforest. No association between histo-blood group antigen expression and sapovirus infection was observed.
RESUMO
OBJECTIVES: To verify the frequency of viruses causing acute gastroenteritis (AGE) in association with the histo-blood group antigen (HBGA) and Rotarix™ vaccination coverage in children from the Amazon region. DESIGN: Fecal and saliva samples were collected from children with AGE (n = 485) and acute respiratory infection (ARI) (n = 249) clinical symptoms. Rotavirus A (RVA), norovirus, human adenovirus (HAdV), and sapovirus (SaV) were verified in feces by molecular detection. Saliva samples were used for HBGA phenotyping/FUT3 genotyping. Blood group types, clinical aspects and Rotarix™ RVA vaccination data were recorded. RESULTS: Norovirus remained the most prevalently detected cause of AGE (38%, 184/485 and ARI 21.3%, 53/249). High HAdV frequencies were observed in AGE children (28.6%, 139/485) and ARI children (37.3%, 93/249). RVA was the third most prevalent virus causing AGE (22.7%, 110/485 and ARI 19.3%, 48/249) and a low RV1 coverage (61%, 448/734) was verified. The SaV frequencies were lower (7.2%, 35/485 for AGE and 6.8%, 17/249 for ARI). Secretor children were HBGA susceptible to HAdV infection (OR 1.5, 95% CI 1.0-2.3; P = 0.04) but not to RVA, norovirus or SaV infection. CONCLUSIONS: Norovirus could be considered the main etiological agent of AGE. No association was verified for HBGA susceptibility to RVA, norovirus and SaV. Secretor children showed a slight susceptibility to HAdV infection and the Le (a-b-) heterogeneous SNPs on the FUT3 gene.
Assuntos
Gastroenterite/virologia , Viroses/epidemiologia , Doença Aguda , Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/isolamento & purificação , Antígenos de Grupos Sanguíneos/análise , Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Pré-Escolar , Fezes/virologia , Feminino , Fucosiltransferases/genética , Gastroenterite/epidemiologia , Gastroenterite/genética , Genótipo , Humanos , Lactente , Masculino , Norovirus/isolamento & purificação , Polimorfismo de Nucleotídeo Único , Infecções Respiratórias/virologia , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Vacinas contra Rotavirus , Saliva , Sapovirus/isolamento & purificação , América do Sul/epidemiologia , Vacinas AtenuadasRESUMO
Human astrovirus (HAstV) 1-8 and highly divergent HAstVMLB1-3 genotypes have been detected in children both with and without acute gastroenteritis (AGE). One hundred and seventy fecal samples from children (≤5 years old) living in the Amazon region were evaluated for the presence of HAstV1-8, HAstV MLB1-3 and HAstVVA1-3, using an usual RT-PCR protocol and a new protocol with specific primers designed to detect HAstVMLB1-3. HAstVMLB1 and HAstV MLB2, as well as the HAstV3 and 5 genotypes were detected. HAstVMLB1-2 genotype was detected for the first time in Brazil at a frequency of 3.5% (6/170).
Assuntos
Infecções por Astroviridae , Gastroenterite , Mamastrovirus , Infecções por Astroviridae/diagnóstico , Infecções por Astroviridae/epidemiologia , Brasil , Criança , Fezes , Gastroenterite/diagnóstico , Genótipo , Humanos , Lactente , Mamastrovirus/genética , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVES: This study aimed to verify the frequency, genotypes, and etiological role of Human Bocavirus (HBoV) in younger Amazonian children with either acute gastroenteritis (AGE) or respiratory infections (ARI). The influence of Rotarix™ vaccination and co-infection status was also investigated. DESIGN: HBoV quantitative polymerase chain reaction (qPCR) testing was done on both fecal and saliva (1468 samples) from 734 children < 5 months old living in the Amazon (Brazil, Guyana, and Venezuela). High and median HBoV viral load samples were used for extraction, nested PCR amplification, and sequencing for genotyping. HBoV mRNA detection was done by reverse transcription following DNA amplification. RESULTS: The overall HBoV frequencies were 14.2% (69/485; AGE) and 14.1% (35/249; ARI) (p = 0.83). HBoV exclusively infected 4.5% (22/485; AGE) and 4% (10/249) of the Amazonian children (Odds ratios 1.13, 95% confidence interval= 2.42-0.52). HBoV 1 was mainly detected in feces and saliva from AGE children; and HBoV2, from ARI children. HBoV mRNA was detected only in feces. The Rotarix™ vaccination status did not affect the HBoV frequencies. CONCLUSIONS: We suggest that, after entry into the air/oral pathways, HBoV1 continues infecting toward the intestinal tract causing AGE. HBoV2 can be a causative agent of AGE and ARI in younger Amazonian children.
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Gastroenterite/virologia , Bocavirus Humano/genética , Infecções por Parvoviridae/virologia , Infecções Respiratórias/virologia , Doença Aguda , Brasil , Coinfecção/virologia , Fezes/virologia , Feminino , Genótipo , Guiana , Bocavirus Humano/isolamento & purificação , Humanos , Lactente , Masculino , Reação em Cadeia da Polimerase , Saliva/virologia , Venezuela , Carga ViralRESUMO
Neste estudo avaliamos o impacto das doenças diarreicas agudas (DDA) associadas aos rotavírus A (RVA), norovírus, adenovírus (HAdV), sapovírus (SaV), bocavírus (HBoV) e astrovírus (HAstV) em crianças ≤ 5 anos atendidas na emergência do Hospital da Criança de Santo Antônio (HCSA), Boa Vista, Roraima (RR), no período de outubro de 2016 a outubro de 2017. Foram coletadas, paralelamente, amostras de fezes e de saliva de 734 crianças sendo 485 com DDA e 249 com infecção respiratória aguda (IRA, grupo controle). O estudo teve a aprovação do CEP:1.333.480, 23/11/2015, UFRR. Os RVA, norovírus, HAdV, SaV e HBoV foram pesquisados nas 734 amostras de fezes e HAstV em 170 amostras de crianças com DDA. Adicionalmente, a presença dos HBoV foi também investigada em 38 amostras de saliva sendo 25 de crianças com DDA e 13 IRA. O perfil de susceptibilidade AB0, Lewis e secretor dos antígenos do grupo histosanguíneo (HBGA) foi determinado para todas as 734 crianças em salivas pela fenotipagem e em amostras de salivas pela genotipagem, sendo para o gene FUT2 em 166 amostras e para FUT3 em 42 amostras. Os aspectos clínicos, epidemiológicos e a cobertura da vacina Rotarix® (RV1) foram também avaliados. Os RVA, norovírus e SaV foram investigados pela metodologia de transcrição reversa seguida de amplificação genômica quantitativa (RT-qPCR); os HBoV e HAdV pela amplificação genômica quantitativa (qPCR) e os HAstV por amplificação genômica qualitativa (RT-PCR). A genotipagem dos HBoV e a detecção/genotipagem dos HAstV foi realizada pela PCR seguida de sequenciamento nucleotídico (método Sanger). O Ensaio imunoenzimático (EIA) e a amplificação genômica específica (PCR-touchdown), seguida de sequenciamento nucleotídico (Sanger) foram utilizadas respectivamente para a fenotipagem e genotipagem dos HBGA.
Nas crianças com DDA (n=485), observou-se as seguintes frequências virais: RVA (22,7%), norovírus (38%), HAdV (33,6%), SaV (7,3%) e HBoV (14,2%). Nas crianças com IRA (n=249), observou as seguintes frequências: RVA (19,3%), norovírus (21,3%), HAdV (39,5%), SaV (5,6%) e HBoV (14,1%). O perfil de detecção do HBoV nas 76 amostras de saliva e fezes pareadas foi diferente e correlacionado com os genótipos 1 a 3 detectados. Quanto aos HAstV, nas 170 amostras investigadas, observou-se as seguintes frequências e genótipos: HAstV clássicos: HAstV3 (0,60%) e HAsV5 (1,8%); HAstV não clássicos: HAstVMLB1-2 (3,5%), sendo esta a primeira descrição do HAstVMLB2 no Brasil. A cobertura vacinal para RV1 calculada foi de 61% e crianças vacinadas na faixa etária entre 6 e 24 meses apresentaram frequências mais elevadas de infecção pelo RVA. As 734 crianças apresentaram majoritariamente (54.5%) o perfil Lea+b+ (fraco secretor) e polimorfirmos de nucleotídico único (SNPs) foram detectados nos genes FUT2 e FUT3. Foi detectada susceptibilidade (HBGA) para a infecção pelos HAdV em crianças com IRA, perfil fraco secretor e grupo sanguíneo A ou O, de acordo com valores de Odds Ratio (OR) calculado. As frequência dos vírus detectados principalmente para os norovírus e HAdV em crianças com DDA, demostram a importância da etiologia viral nas DDA em crianças ≤ 5 anos de idade no período do estudo e podem estar relacionadas ao fator de susceptibilidade ao HBGA (incluindo heterogeneidade genética) e a baixa cobertura de RV1. (AU)
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Humanos , Pré-Escolar , Infecções por Rotavirus , Adenovírus Humanos , Pré-Escolar , Infecções por Caliciviridae , Sapovirus , Disenteria , AvastrovirusRESUMO
Sexual maturation and reproduction influence the status of a number of physiological processes and consequently the ecology and behaviour of cephalopods. Using Octopus mimus as a study model, the present work was focused in the changes in biochemical compound and activity that take place during gonadal maturation of females and its consequences in embryo and hatchlings characteristics. To do that, a total of 31 adult females of O. mimus were sampled to follow metabolites (ovaries and digestive gland) and digestive enzyme activities (alkaline and acidic proteases) during physiological and functional maturation. Levels of protein (Prot), triacylglyceride (TG), cholesterol (Chol), glucose (Glu), and glycogen (Gly) were evaluated. Groups of eggs coming from mature females were also sampled along development and after hatching (paralarvae of 1 and 3 days old) to track metabolites (Prot, TG, Glu, Gly, TG, Chol), digestive enzymes activity (Lipase, alkaline proteases, and acidic proteases), and antioxidant/detoxification defence indicators with embryos development. Based on the data obtained, we hypothesized that immature females store Chol in their ovaries, probably from the food they ingested, but switch to TG reserves at the beginning of the maturation processes. At the same time, results suggest that these processes were energetically supported by Glu, obtained probably from Gly breakdown by gluconeogenic pathways. Also, was observed that embryos metabolites and enzyme activities (digestive and antioxidant/detoxification enzymes) where maintained without significant changes and in a low activity during the whole organogenesis, meaning that organogenesis is relatively not energetically costly. In contrast, after organogenesis, a mobilization of nutrients and activation of the metabolic and digestive enzymes was observed, together with increments in consumption of yolk and Gly, and reduction in lipid peroxidation. Derived from our results, we also have the hypothesis that reactive oxygen species (ROS) were produced during the metabolic processes that occurs in ovarian maturation. Those ROS may be in part transferred to the egg provoking a ROS charge to the embryos. The elimination of ROS in embryos started when the activity of the heart and the absorption of the yolk around stages XIV and XV were evident. Altogether, these processes allowed the paralarvae to hatch with buffered levels of ROS and with the antioxidant defence mechanisms ready to support further ROS production derived from paralarvae higher life stage requirements (feeding and metabolic demands).
RESUMO
The Histo-blood group antigens (HBGA) are host genetic factors associated with susceptibility to rotavirus (RV) and human norovirus (HuNoV), the major etiological agents of viral acute gastroenteritis (AGE) worldwide. The FUT2 gene expressing the alpha-1, 2-L- fucosyltransferase enzyme is important for gut HBGA expression, and also provides a composition of the phenotypic profile achieved through mutations occurring in populations with different evolutionary histories; as such, it can be considered a genetic population marker. In this study, Lewis and secretor HBGA phenotyping was performed using 352 saliva samples collected from children between three months and five years old born in the Amazon (Brazil, Venezuela and English Guyana) presenting AGE or acute respiratory infection (ARI), the latter considered as control samples. The total of children phenotyped as secretors was 323, corresponding to 91.80%. From these, 207 (58.80%) had a Le (aâ¯+â¯b+) profile. The HBGA profiles were equally found in children with AGE as well as with ARI. The rs1047781 of the FUT2 gene was not detected in DNA from saliva cells with a Le (a+b+) profile. However, mutations not yet described in the FUT2 gene were observed: missense 325A>T, 501C>T, 585C>T, 855A>T and missense substitutions 327C>T [S (Ser) > C (Cys)], 446 T>C [L(Leu) > P(Pro)], 723C>A [N(Asn) > K(Lys)], 724A>T [I(Ile) > F(Phe)], 736C>A [H(His) > N(Asn)]. The SNP distribution in the FUT2 gene of the analyzed samples was very similar to that described in Asian populations, including indigenous tribes.