RESUMO
Acute gastroenteritis (AGE) is a common pediatric infection that remains a significant cause of childhood morbidity and mortality worldwide, especially in low-income regions. Thus, the objective of this study was to detect human adenovirus (HAdV) and non-polio enterovirus (NPEV) in fecal samples from the Gastroenteritis Surveillance Network, and to identify circulating strains by nucleotide sequencing. A total of 801 fecal samples were tested using qPCR/RT-qPCR, and 657 (82.0%) were inoculated into HEp-2C and RD cell lines. The HAdV and NPEV positivity rates obtained using qPCR/RT-qPCR were 31.7% (254/801) and 10.5% (84/801), respectively, with 5.4% (43/801) co-detection. Cytopathic effect was observed in 9.6% (63/657) of patients, 2.7% (18/657) associated with HAdV, and 6.2% (41/657) associated with NPEV after testing by ICC-PCR. A comparison of the two methodologies demonstrated an agreement of 93.5% for EVNP and 64.4% for HAdV. These two viruses were detected throughout the study period, with HAdV positivity rates ranging from 41% in Amapá to 18% in Pará. The NEPV varied from 18% in Pará/Rondônia to 3% in Acre. The most affected age group was over 60 months for both HAdV and NPEV. Samples previously positive for rotavirus and norovirus, which did not show a major difference in the presence or absence of diarrhea, fever, and vomiting, were excluded from the clinical analyses of these two viruses. These viruses circulated over five years, with a few months of absence, mainly during the months corresponding to the waves of SARS-CoV-2 infection in Brazil. Five HAdV species were identified (A, B, C, D, and F), with a greater predominance of HAdV-F41 (56.5%) followed by HAdV-C (15.2%). Three NPEV species (A, B, and C) were detected, with serotypes E14 (19.3%) and CVA-24 (16.1%) being the most prevalent. The present study revealed a high diversity of NPEV and HAdV types circulating in children with AGE symptoms in the northern region of Brazil.
Assuntos
Adenovírus Humanos , Enterovirus , Fezes , Gastroenterite , Humanos , Gastroenterite/virologia , Gastroenterite/epidemiologia , Brasil/epidemiologia , Fezes/virologia , Pré-Escolar , Lactente , Adenovírus Humanos/genética , Adenovírus Humanos/isolamento & purificação , Adenovírus Humanos/classificação , Masculino , Enterovirus/genética , Enterovirus/isolamento & purificação , Feminino , Criança , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Infecções por Enterovirus/diagnóstico , Doença Aguda , Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/virologia , FilogeniaRESUMO
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.
Assuntos
Infecções por Adenovirus Humanos/epidemiologia , Adenovírus Humanos/classificação , Adenovírus Humanos/genética , Gastroenterite/epidemiologia , Adenovírus Humanos/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , Criança , Pré-Escolar , DNA Viral/genética , Fezes/virologia , Feminino , Gastroenterite/virologia , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Estudos Retrospectivos , Adulto JovemRESUMO
Human enteric adenovirus species F (HAdV-F) is one of the most common pathogens responsible for acute gastroenteritis worldwide. Brazil is a country with continental dimensions where continuous multiregional surveillance is vital to establish a more complete picture of the epidemiology of HAdV-F. The aim of the current study was to investigate the molecular epidemiology of HAdV-F using full-genome data in rural and low-income urban areas in northern Brazil. This will allow a genetic comparison between Brazilian and global HAdV-F strains. The frequency of HAdV-F infections in patients with gastroenteritis and molecular typing of positive samples within this period was also analysed. A total of 251 stool samples collected between 2010 and 2016 from patients with acute gastroenteritis were screened for HAdV-F using next-generation sequencing techniques. HAdV-F infection was detected in 57.8â% (145/251) of samples. A total of 137 positive samples belonged to HAdV-F41 and 7 to HAdV-F40. HAdV-F40/41 dual infection was found in one sample. Detection rates did not vary significantly according to the year. Single HAdV-F infections were detected in 21.9â% (55/251) of samples and mixed infections in 37.4â% (94/251), with RVA/HAdV-F being the most frequent association (21.5â%; 54/251). Genetic analysis indicated that the HAdV-F strains circulating in Brazil were closely related to worldwide strains, and the existence of some temporal order was not observed. This is the first large-scale HAdV-F study in Brazil in which whole-genome data and DNA sequence analyses were used to characterize HAdV-F strains. Expanding the viral genome database could improve overall genotyping success and assist the National Center for Biotechnology Information (NCBI)/GenBank in standardizing the HAdV genome records by providing a large set of annotated HAdV-F genomes.
Assuntos
Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/genética , Gastroenterite/virologia , Variação Genética , Adenovírus Humanos/classificação , Adenovírus Humanos/isolamento & purificação , Adolescente , Adulto , Idoso , Brasil/epidemiologia , Criança , Pré-Escolar , Biologia Computacional , Estudos Transversais , Fezes/virologia , Feminino , Gastroenterite/epidemiologia , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Masculino , Metagenômica , Pessoa de Meia-Idade , Epidemiologia Molecular , Tipagem Molecular , Filogenia , Recombinação Genética , Estudos Retrospectivos , Análise de Sequência de DNA , Adulto JovemRESUMO
Leachate from solid waste landfill is a dark liquid of variable composition and possible source of contamination of groundwater and surface waters. This study aims to assess skimmed milk flocculation and ultracentrifugation as viral concentration methods associated to different nucleic acid extraction protocols in order to establish a methodology for virus recovery from sanitary landfill leachate. Spiking experiments using human adenovirus (HAdV) and bacteriophage PP7 revealed the association of QIAamp Fast DNA Stool mini kit® nucleic acid extraction and ultracentrifugation as an effective method for recovering HAdV (346.18%) and PP7 (523.97%) when compared to organic flocculation method (162.64% for HAdV and 0.61% for PP7) that presented PCR inhibition in all undiluted samples. Ultracentrifugation applied in three landfill samples confirm efficiency of the methodology detecting HAdV in all samples with a mean of 3.44E + 06 ± 1.56E + 06 genomic copies/mL. Nucleotide sequencing characterized HAdV as belonging to group B and F. JC polyomavirus (JCPyV) was also investigated in those samples; however, detection was not observed. Methodologies for detection of viruses in leachate can be useful to generate data for future health risk analysis of workers who have contact with solid urban waste, as well as populations exposed to different environmental matrices contaminated by these effluents.
Assuntos
Adenovírus Humanos/isolamento & purificação , Bacteriófagos/isolamento & purificação , Virologia/métodos , Poluentes Químicos da Água/análise , Adenovírus Humanos/classificação , Adenovírus Humanos/genética , Bacteriófagos/classificação , Bacteriófagos/genética , Monitoramento Ambiental , Dosagem de Genes , Genoma Viral , Humanos , Reação em Cadeia da Polimerase , Resíduos Sólidos/análiseRESUMO
Human Adenovirus species C (HAdV-C) is the most common etiologic agent of respiratory disease. In the present study, we characterized the nearly full-length genome of one potential new HAdV-C recombinant strain constituted by Penton and Fiber proteins belonging to type 89 and a chimeric Hexon protein of types 1 and 89. By using viral metagenomics techniques, we screened out, in the states of Tocantins and Pará, Northern and North regions of Brazil, from 2010 to 2016, 251 fecal samples of children between 0.5 to 2.5 years old. These children were presenting acute diarrhea not associated with common pathogens (i.e., rotavirus, norovirus). We identified two HAdV-C strains in two distinct patients. Phylogenetic analysis performed using all complete genomes available at GenBank database indicated that one strain (HAdV-C BR-245) belonged to type 1. The phylogenetic analysis also indicated that the second strain (HAdV-C BR-211) was located at the base of the clade formed by the newly HAdV-C strains type 89. Recombination analysis revealed that strain HAdV-C BR-211 is a chimera in which the variable regions of Hexon gene combined HAdV-C1 and HAdV-C89 sequences. Therefore, HAdV-C BR-211 strain possesses a genomic backbone of type HAdV-C89 and a unique insertion of HAdV-C1 in the Hexon sequence. Recombination may play an important driving force in HAdV-C diversity and evolution. Studies employing complete genomic sequencing on circulating HAdV-C strains in Brazil are needed to understand the clinical significance of the presented data.
Assuntos
Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/genética , Genoma Viral , Adenovírus Humanos/classificação , Adenovírus Humanos/isolamento & purificação , Sequência de Aminoácidos , Brasil , Proteínas do Capsídeo/genética , Evolução Molecular , Genômica , Filogenia , Recombinação GenéticaRESUMO
The availability of drinking water is one of the main determinants of quality of life, disease prevention and the promotion of health. Viruses are important agents of waterborne diseases and have been described as important markers of human faecal contamination. This study aimed to investigate viruses' presence as an indicator of drinking water quality in low-income communities in the Manguinhos area, Rio de Janeiro, Brazil. Three hundred and four drinking water samples (2L/each) were collected along the drinking water distribution-to-consumption pathway in households, as well as healthcare and school units. Water samples were collected both directly from the water supply prior to distribution and after storage in tanks and filtration units. Using qPCR, viruses were detected 50 times in 45 water samples (15%), 19 of these being human adenovirus, 17 rotavirus A and 14 norovirus GII. Viral loads recovered ranged from 5E+10 to 8.7E+106 genome copies/Liter. Co-detection was observed in five household water samples and there was no difference regarding virus detection across sampling sites. Precarious and inadequate environmental conditions characterized by the lack of local infrastructure regarding basic sanitation and waste collection in the territory, as well as negligent hygiene habits, could explain viral detection in drinking water in regions with a water supply system.
Assuntos
Adenovírus Humanos/isolamento & purificação , Água Potável/virologia , Gastroenterite/virologia , Norovirus/isolamento & purificação , Rotavirus/isolamento & purificação , Adenovírus Humanos/classificação , Adenovírus Humanos/genética , Adenovírus Humanos/crescimento & desenvolvimento , Brasil/epidemiologia , Água Potável/análise , Gastroenterite/economia , Gastroenterite/epidemiologia , Humanos , Higiene , Norovirus/classificação , Norovirus/genética , Norovirus/crescimento & desenvolvimento , Pobreza , Qualidade de Vida , Características de Residência/estatística & dados numéricos , Rotavirus/classificação , Rotavirus/genética , Rotavirus/crescimento & desenvolvimento , Qualidade da Água , Abastecimento de Água/economiaRESUMO
AIMS: To determine the seasonal occurrence and diversity of norovirus (NoV) and human adenovirus (HAdV) in groundwater from sinkholes, and brackish water used for recreational activities in the karst aquifer of the Yucatan Peninsula, Mexico. METHODS AND RESULTS: Hollow fibre ultrafiltration was used to concentrate viruses and standard plaque assay methods were used to enumerate somatic and F+ specific coliphages as viral indicators. Real-time quantitative polymerase chain reaction assays were used to estimate the number of genome copies for NoV strains GI, and GII, and HAdVs. The predominant NoV genotypes and HAdV serotypes were identified by comparative sequence analysis. Somatic and male F+ specific coliphages were detected at concentrations up to 94 and 60 plaque-forming units per 100 ml respectively. The NoV genogroup I (GI) was associated with 50% of the sampled sites during the rainy season only, at concentrations ranging from 120 to 1600 genome copies per litre (GC l-1 ). The NoV genogroup II (GII) was detected in 30 and 40% of the sampled sites during the rainy and dry seasons, respectively, at concentrations ranging from 10 to 290 GC l-1 . During the rainy and dry seasons, HAdVs were detected in 20% of the sites, at concentrations ranging from 24 to 690 GC l-1 . Identification of viral types revealed the presence of NoV GI.2, GII.Pe, GII.P16 and GII.P17, and HAdV F serotypes 40 and 41. CONCLUSIONS: These findings demonstrate that NoVs and HAdVs are prevalent as virus contaminants in the karst aquifer, representing potential health risks particularly during the rainy season, in one of the most important areas used for tourism in Mexico. SIGNIFICANCE AND IMPACT OF THE STUDY: This is one of the few studies conducted in karst aquifers that provide a foundational baseline of the distribution, concentrations and diversity of NoVs and HadVs in these particular environments.
Assuntos
Adenovírus Humanos , Água Subterrânea/virologia , Norovirus , Adenovírus Humanos/classificação , Adenovírus Humanos/genética , Adenovírus Humanos/isolamento & purificação , México , Norovirus/classificação , Norovirus/genética , Norovirus/isolamento & purificação , Microbiologia da ÁguaRESUMO
Human adenovirus (HAdV) has been recognized as a significant viral pathogen implicated in neurological diseases, particularly in immunocompromised patients. However, its involvement in meningoencephalitis remains unclear. The aim of this study was to investigate HAdV and other viral co-infections in the cerebrospinal fluid (CSF) of patients suspected of having either meningoencephalitis or encephalitis. A total of 373 CSF samples from patients under clinical suspicion of neurological viral infection were included in this study. HAdV was investigated by conventional or multiplex real-time PCR, for different time periods. The frequency of HAdV central nervous system (CNS) infection was 1.08%, predominating in female patients with a predisposing condition, and presented with HAdV encephalitis. HAdV CNS infection was found to occur during the months of autumn and winter. The frequency of HAdV detected in CSF positive samples increased after the change in the diagnostic method from conventional to multiplex real-time PCR. There were no specific NMRI or EEG characteristics and two CSF samples with HAdV encephalitis had normal CSF WBC count. There were two cases of co-infection with HIV; no other co-infections with enterovirus or herpes family viruses were detected. All patients had good outcome. Although HAdV is rarely observable in CNS infectious syndromes, it must be investigated particularly in immunocompromised patients.
Assuntos
Adenovírus Humanos/genética , DNA Viral/genética , Encefalite Viral/diagnóstico , Meningite Viral/diagnóstico , Meningoencefalite/diagnóstico , Adenovírus Humanos/classificação , Adenovírus Humanos/isolamento & purificação , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , DNA Viral/líquido cefalorraquidiano , Encefalite Viral/líquido cefalorraquidiano , Encefalite Viral/patologia , Encefalite Viral/virologia , Feminino , Humanos , Contagem de Leucócitos , Leucócitos/virologia , Masculino , Meningite Viral/líquido cefalorraquidiano , Meningite Viral/patologia , Meningite Viral/virologia , Meningoencefalite/líquido cefalorraquidiano , Meningoencefalite/patologia , Meningoencefalite/virologia , Pessoa de Meia-Idade , Estudos Prospectivos , Estações do Ano , Carga ViralRESUMO
The role of human adenovirus (HAdV) infection in different acute diseases, such as febrile exudative tonsillitis, conjunctivitis, and pharyngoconjunctival fever is well established. However, the relationships, if any, of HAdV persistence and reactivation in the development of the chronic adenotonsillar disease is not fully understood. The present paper reports a 3-year cross-sectional hospital-based study aimed at detecting and quantifying HAdV DNA and mRNA of the HAdV hexon gene in adenoid and palatine tonsil tissues and nasopharyngeal secretions (NPS) from patients with adenotonsillar hypertrophy or recurrent adenotonsillitis. HAdV C, B, and E were detectable in nearly 50% of the patients, with no association with the severity of airway obstruction, nor with the presence of recurrent tonsillitis, sleep apnea or otitis media with effusion (OME). Despite the higher rates of respiratory viral coinfections in patients with HAdV, the presence of other viruses, including DNA and RNA viruses, had no association with HAdV replication or shedding in secretions. Higher HAdV loads in adenoids showed a significant positive correlation with the presence of sleep apnea and the absence of OME. Although this study indicates that a significant proportion (~85%) of individuals with chronic adenotonsillar diseases have persistent nonproductive HAdV infection, including those by HAdV C, B, and E, epithelial and subepithelial cells in tonsils seem to be critical for HAdV C production and shedding in NPS in some patients, since viral antigen was detected in these regions by immunohistochemistry in four patients, all of which were also positive for HAdV mRNA detection.
Assuntos
Tonsila Faríngea/virologia , Infecções por Adenovirus Humanos/virologia , Tonsila Palatina/virologia , Replicação Viral , Tonsila Faríngea/patologia , Infecções por Adenovirus Humanos/diagnóstico , Adenovírus Humanos/classificação , Adenovírus Humanos/isolamento & purificação , Adenovírus Humanos/fisiologia , Adolescente , Criança , Pré-Escolar , Estudos Transversais , DNA Viral/isolamento & purificação , Feminino , Humanos , Hipertrofia , Lactente , Masculino , Tonsila Palatina/patologia , Tonsilite/virologiaRESUMO
BACKGROUND: In Brazil, limited data are available regarding the incidence and molecular epidemiology of Human Adenovirus (HAdV) in diarrheic disease, especially in the post rotavirus (RVA) vaccine era. OBJECTIVE: The aims of the study were to investigate the frequency of HAdV infections in patients with gastroenteritis during a 6-year period (2012-2017); conduct molecular typing of positive strains, and obtain further information on the HAdV seasonality. STUDY DESIGN: A total of 3003 fecal samples negative for both, RVA and Norovirus, were selected and tested for HAdV by PCR. Positive HAdV samples were sequenced to obtain genotype identification. RESULTS: HAdV was detected in 3.9% (117/3003); 76 belong to species F (70 HAdV-F41; 6 HAdVF40), 31 to species C (15 HAdV-C1; 13 HAdV-C2; 3 HAdV-C5), 5 belong to species D (3 HAdVD56 and 2 untyped), 4 belong to species A (2 HAdV-A12; 2 HAdV-A31), and 1 belong to HAdVB3. Detection rate significantly varied according to the year, suggesting that HAdV infections show a tendency to occur in natural oscillatory fluctuation. No consistent seasonal pattern was identified. Children ≤5 years exhibited higher positivity rate, reinforcing that HAdV is an important pathogen in childhood diarrhea. Genetic analysis indicated that HAdV strains circulating in Brazil were closely related to worldwide strains, and there is no evidence for the introduction of a particular HAdV variant in the country. CONCLUSIONS: The present investigation does not suggest that HAdV has assumed an epidemiological importance in Brazil after the RVA vaccine introduction and contributed to the definition of the clinical and public health significance of HAdV infections.