Norovirus (NoV), sapovirus (SaV) and human astrovirus (HAstV) are viral pathogens that are associated with outbreaks and sporadic cases of gastroenteritis. However, little is known about the occurrence of these pathogens in relatively isolated communities, such as the remnants of African-descendant villages (“Quilombola”). The objective of this study was the frequency determination of these viruses in children under 10 years, with and without gastroenteritis, from a “Quilombola” Community, Northern Brazil. A total of 159 stool samples were obtained from April/2008 to July/2010 and tested by an enzyme immunoassay (EIA) and reverse transcription-polymerase chain reaction (RT-PCR) to detect NoV, SaV and HAstV, and further molecular characterization was performed. These viruses were detected only in the diarrheic group. NoV was the most frequent viral agent detected (19.7%-16/81), followed by SaV (2.5%-2/81) and HAstV (1.2%-1/81). Of the 16 NoV-positive samples, 14 were sequenced with primers targeting the B region of the polymerase (ORF1) and the D region of the capsid (ORF2). The results showed a broad genetic diversity of NoV, with 12 strains being classified as GII-4 (5–41.7%), GII-6 (3–25%), GII-7 (2–16.7%), GII-17 (1–8.3%) and GI-2 (1–8.3%), as based on the polymerase region; 12 samples were classified, based on the capsid region, as GII-4 (6–50%, being 3–2006b variant and 3–2010 variant), GII-6 (3–25%), GII-17 (2–16.7%) and GII-20 (1–8.3%). One NoV-strain showed dual genotype specificity, based on the polymerase and capsid region (GII-7/GII-20). This study provides, for the first time, epidemiological and molecular information on the circulation of NoV, SaV and HAstV in African-descendant communities in Northern Brazil and identifies NoV genotypes that were different from those detected previously in studies conducted in the urban area of Belém. It remains to be determined why a broader NoV diversity was observed in such a semi-isolated community.
BACKGROUND Norovirus (NoV) is a major cause of acute gastroenteritis (AGE) worldwide, especially in children under five years. Studies involving the detection and molecular characterisation of NoV have been performed in Brazil, demonstrating its importance as an etiological agent of AGE.OBJECTIVES The objectives of this study were to investigate the frequency of human NoV and to genotype the strains isolated from 0-14-year-old patients of AGE in Manaus, Brazil, over a period of two years.METHODS A total of 426 faecal samples were collected between January 2010 and December 2011. All samples were tested for the presence of NoV antigens using a commercial enzyme immunoassay kit. RNA was extracted from all faecal suspensions and reverse transcription-polymerase chain reaction (RT-PCR) for the NoV-polymerase partial region was performed as a trial test. Positive samples were then subjected to PCR with specific primers for partial capsid genes, which were then sequenced.FINDINGS NoV was detected in 150 (35.2%) faecal samples, for at least one of the two techniques used. NoV was detected in children from all age groups, with the highest positivity observed among the group of 1-2 years old. Clinically, fever was verified in 43% of the positive cases and 46.3% of the negative cases, and vomiting was observed in 75.8% and 70.8% cases in these groups, respectively. Monthly distribution showed that the highest positivity was observed in January 2010 (81.2%), followed by February and April 2010 and March 2011, when the positivity rate reached almost 50%. Phylogenetic analyses performed with 65 positive strains demonstrated that 58 (89.2%) cases of NoV belonged to genotype GII.4, five (7.7%) to GII.6, and one (1.5%) each to GII.7 and GII.3.MAIN CONCLUSIONS This research revealed a high circulation of NoV GII.4 in Manaus and contributed to the understanding of the importance of this virus in the aetiology of AGE cases, especially in a region with such few studies available.
SUMMARYSapoviruses (SaVs) are responsible for acute gastroenteritis in humans, especially children and the elderly. In Brazil, data on SaVs infections are very limited, especially in Northern Brazil. Here, we investigated the occurrence of SaVs in samples from hospitalized children under ten years old that presented acute gastroenteritis. Positive samples were genotyped and phylogenetic analysis was performed using prototype strains sequences obtained from GenBank database. In total, 156 fecal samples were screened by RT-PCR for SaVs. A positivity rate of 3.8% (6/156) was found in children under three years of age. Four genotypes were detected: GI.I, GI.2 and GII.2?-GII.4?/GII.4, suggesting a possible inter-genotypes recombination. Most infections (83.3%) occurred between August and September. The positivity was similar to that found in other countries and genotyping demonstrated the presence of distinct genotypes. To our knowledge, this is the first study reporting the circulation of SaVs in Manaus, state of Amazonas, Amazon region, Brazil.
Sapoviruses (SaVs) belong to the family Caliciviridae and are related to gastroenteritis viruses of humans and animals. These agents have been reported from several countries of the world and represent an important cause of economic loss. The Amazon area has a high degree of diversity of animals and plants, is located in the Northern Region of Brazil and accounts for a large part of the Brazilian territory. In this study, stool samples were collected from pigs during the phase of nursing (less than 28 days of age) and post-weaning (29 to 56 days of age) from January 2008 to February 2009. A total of 169 specimens (108 nursing and 61 post-weaning pigs) were tested by reverse transcription polymerase chain reaction (RT-PCR) using the primers p289/p290 for the detection of caliciviruses (CVs), i.e., SaVs and noroviruses (NoVs). Positive sequences were analyzed using BioEdit software (v. 7.1.3.0) and compared with other sequences registered in the GenBank database. A positive frequency of 12.4 % (21/169) was observed, and all of the viruses found were identified as SaVs, with 15 belonging to genogroup GIII (71.4 %), three to GVII-1 (14.3 %) and three to GVIII-2 (14.3 %). No NoVs were detected. The frequency of SaV infections was significantly higher in nursing pigs (17.6 %-19/108) than in post-weaning pigs (3.3 %-2/61). Considering the consistency of the samples, 14.7 % of the samples were classified as diarrheic, but statistical analysis demonstrated that there was no significant difference compared to normal specimens (p = 0.5795). For the first time, we have demonstrated the circulation of SaVs in pigs from the Amazon.
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