RESUMO
The Adenoviridae family is composed by a high diversity of viruses that are extremely resistant in environment and are frequently excreted in animal reservoir feces for long periods. The knowledge of adenovirus (AdV) diversity among wild species may be important for the understanding of the epidemiology of putative emerging diseases. Cavia aperea aperea, commonly known as wild guinea pigs, wild cavies, or preas, are small herbivorous rodents widely distributed throughout South America and classified in Caviidae family, as well as domestic guinea pigs and capybaras. In order to investigate their potential role as reservoir of zoonotic agents, the present study aimed to verify the presence of AdV in fecal samples of 14 preas from Northeast Brazil. When submitted to nested PCR, two out of 14 samples (14.28%) were positive for AdV and classified as human Mastadenovirus C (HAdV-C) using DNA sequencing and phylogenetic analysis. Wild guinea pigs are synanthropic rodents that live in close contact with humans. The investigation of viral agents in rodents is important due to their potential role as reservoirs of human and animal pathogens. Moreover, the present work presents the first known evidence of HAdV in wild guinea pig stool samples, which may represent both the impact of anthropogenic pollution to wild animals and an important knowledge in terms of human health.
Assuntos
Animais Selvagens , Mastadenovirus , Humanos , Cobaias , Animais , Filogenia , Fezes , Análise de Sequência de DNA , Mastadenovirus/genéticaRESUMO
Molecular methodologies providing data on viral concentration and infectivity have been successfully used in environmental virology, supporting quantitative risk assessment studies. The present study aimed to assess human mastadenovirus (HAdV) intact particles using a derivative of propidium monoazide associated with qPCR (PMAxx-qPCR) in aquatic matrices. Initially, different concentrations of PMAxx were evaluated to establish an optimal protocol for treating different naturally contaminated matrices, using 10 min incubation in the dark at 200 rpm at room temperature and 15 min of photoactivation in the PMA-Lite™ LED photolysis device. There was no significant reduction in the quantification of infectious HAdV with increasing concentration of PMAxx used (20 µM, 50 µM, and 100 µM), except for sewage samples. In this matrix, a reduction of 5.01 log of genomic copies (GC)/L was observed from the concentration of 50 µM and revealed 100% HAdV particles with damaged capsids. On the other hand, the mean reduction of 0.51 log in stool samples using the same concentration mentioned above demonstrated 83% of damaged particles eliminated in the stool. Following, 50 µM PMAxx-qPCR protocol revealed a log reduction of 0.91, 0.67, and 1.05 in other samples of raw sewage, brackish, and seawater where HAdV concentration reached 1.47 × 104, 6.81 × 102, and 2.33 × 102 GC/L, respectively. Fifty micrometers of PMAxx protocol helped screen intact viruses from different matrices, including sea and brackish water.
Assuntos
Mastadenovirus , Esgotos , Humanos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Água do MarRESUMO
ABSTRACT Human Adenovirus 36 (HAdV-36) has been related to diverse effects on metabolism and may attenuate the lipid accumulation in kidneys with increased adiposity. Some of these effects would be related to viral persistence. However, until now, a model of persistent in vitro infection by HAdV-36 is unknown. In this study, we examined the cells of the Vero lineage to explore their permissiveness to long-term HAdV-36 infection. HAdV-36 was productively replicated in Vero cells and maintained long-term infection for up to 35 cell passages. A subculture was obtained from the cells that survived the primary infection at a low MOI (0.5). The production of the extracellular infectious virus with titers ranging from 104 to 106 TCID50/mL and DNA-bearing cells was detected. In long-term infected cells, the intracellular distribution of viral antigen was demonstrated by performing immunolocalization (IFI) and expression of cell-viral antigen in 50% of cells by flow cytometry, using anti-HAdV-36 hyperimmune rabbit serum. Furthermore, E1a and E4orf1 genes in long-term infected passages showed a decreasing trend. Our preliminary results reveal that renal epithelial monkey cells are permissive for the productive infection of HAdV-36. Vero cell culture long-term infection might be a promising model for addressing the fundamental aspects of the HAdV-36 biology that cannot reveal broadly-used cultures, which do not maintain long-term infection in primary or transformed cells.
RESUMO
A common viral replication strategy is characterized by the assembly of intracellular compartments that concentrate factors needed for viral replication and simultaneously conceal the viral genome from host-defense mechanisms. Recently, various membrane-less virus-induced compartments and cellular organelles have been shown to represent biomolecular condensates (BMCs) that assemble through liquid-liquid phase separation (LLPS). In the present work, we analyze biophysical properties of intranuclear replication compartments (RCs) induced during human adenovirus (HAdV) infection. The viral ssDNA-binding protein (DBP) is a major component of RCs that contains intrinsically disordered and low complexity proline-rich regions, features shared with proteins that drive phase transitions. Using fluorescence recovery after photobleaching (FRAP) and time-lapse studies in living HAdV-infected cells, we show that DBP-positive RCs display properties of liquid BMCs, which can fuse and divide, and eventually form an intranuclear mesh with less fluid-like features. Moreover, the transient expression of DBP recapitulates the assembly and liquid-like properties of RCs in HAdV-infected cells. These results are of relevance as they indicate that DBP may be a scaffold protein for the assembly of HAdV-RCs and should contribute to future studies on the role of BMCs in virus-host cell interactions.
Assuntos
Adenoviridae/metabolismo , Condensados Biomoleculares , Proteínas de Ligação a DNA/metabolismo , Compartimentos de Replicação Viral/fisiologia , Replicação Viral/fisiologia , Adenoviridae/genética , Infecções por Adenoviridae , Adenovírus Humanos/metabolismo , Linhagem Celular , Proteínas de Ligação a DNA/química , Interações entre Hospedeiro e Microrganismos , Humanos , Organelas/virologia , Domínios Proteicos , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/metabolismoRESUMO
With the arrival of coronavirus disease 2019 (COVID-19) in Brazil in February 2020, several preventive measures were taken by the population aiming to avoid severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection including the use of masks, social distancing, and frequent hand washing then, these measures may have contributed to preventing infection also by other respiratory viruses. Our goal was to determine the frequencies of Influenza A and B viruses (FLUAV/FLUBV), human mastadenovirus C (HAdV-C), Enterovirus 68 (EV-68), and rhinovirus (RV) besides SARS-CoV-2 among hospitalized patients suspect of COVID-19 with cases of acute respiratory disease syndrome (ARDS) in the period of March to December 2020 and to detect possible coinfections among them. Nucleic acid detection was performed using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) in respiratory samples using naso-oropharyngeal swabs and bronchoalveolar lavage. A total of 418 samples of the 987 analyzed (42.3%) were positive for SARS-CoV-2, 16 (1.62%) samples were positive for FLUAV, no sample was positive for FLUBV or EV-68, 67 (6.78%) samples were positive for HAdV-C, 55 samples were positive for RV 1/2 (26.3%) and 37 for RV 2/2 (13.6%). Coinfections were also detected, including a triple coinfection with SARS-CoV-2, FLUAV, and HAdV-C. In the present work, a very low frequency of FLUV was reported among hospitalized patients with ARDS compared to the past years, probably due to preventive measures taken to avoid COVID-19 and the high influenza vaccination coverage in the region in which this study was performed.
Assuntos
Infecções por Adenoviridae/epidemiologia , COVID-19/epidemiologia , Resfriado Comum/epidemiologia , Infecções por Enterovirus/epidemiologia , Influenza Humana/epidemiologia , Distanciamento Físico , Infecções por Adenoviridae/prevenção & controle , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , COVID-19/prevenção & controle , Criança , Pré-Escolar , Coinfecção/epidemiologia , Coinfecção/virologia , Resfriado Comum/prevenção & controle , Enterovirus Humano D/genética , Enterovirus Humano D/isolamento & purificação , Infecções por Enterovirus/prevenção & controle , Feminino , Humanos , Lactente , Vírus da Influenza A/genética , Vírus da Influenza A/isolamento & purificação , Vírus da Influenza B/genética , Vírus da Influenza B/isolamento & purificação , Influenza Humana/prevenção & controle , Masculino , Máscaras , Mastadenovirus/genética , Mastadenovirus/isolamento & purificação , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Rhinovirus/genética , Rhinovirus/isolamento & purificação , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Adulto JovemRESUMO
Human adenoviruses (HAdVs) are associated with respiratory infection in the human population worldwide, but HAdV is underreported and less studied than other respiratory viruses. We investigated HAdV in patients with respiratory infection in Rio Grande do Sul (RS), Brazil, between 2004 and 2018. The frequency and seasonality of HAdV, clinical symptoms and underlying diseases were analysed. Respiratory samples from outpatients with acute respiratory illness (ARI) who attended sentinel units and from inpatients with severe acute respiratory infection (SARI) were collected for HAdV detection by immunofluorescence assay; demographic and clinical data were analysed. In total, 43,514 cases of respiratory infection were analysed, of which 8,901 were ARI (20.5%), and 34,613 (79.5%) were SARI. Respiratory viruses were detected in 35.8% of the cases. The frequency of HAdV in relation to respiratory viruses was 2.8%. HAdV circulated year-round, with higher frequency during winter and early spring; increases in the average monthly temperature were associated with decreases in HAdV infections (p = 0.013). Most hospitalized patients with HAdV were male (p = 0.003). HAdV infection showed association with age (p < 0.001), and children between 1 and 5 years old accounted for 30.8% of the outpatients, whereas among cases of SARI, 88.2% were paediatric patients. Among inpatients with HAdV, 3% died, and of these, the majority had at least one underlying condition, such as cardiopathy and immunosuppression. HAdV infection of the respiratory tract causes morbidity and mortality, and individuals with heart diseases and the immunocompromised are at higher risk of fatality.
Assuntos
Infecções por Adenovirus Humanos/diagnóstico , Adenovírus Humanos/isolamento & purificação , Cardiopatias/virologia , Infecções Respiratórias/epidemiologia , Infecções por Adenovirus Humanos/epidemiologia , Brasil/epidemiologia , Criança , Pré-Escolar , Humanos , Lactente , Masculino , Infecções Respiratórias/virologia , Estações do AnoRESUMO
The current SARS-CoV-2 pandemic has imposed new challenges and demands for health systems, especially in the development of new vaccine strategies. Vaccines for many pathogens were developed based on the display of foreign epitopes in the variable regions of the human adenovirus (HAdV) major capsid proteins (hexon, penton and fiber). The humoral immune response against the HAdV major capsid proteins was demonstrated to play a role in the development of an immune response against the epitopes in display. Through the immunoinformatic profiling of the major capsid proteins of HAdVs from different species, we developed a modular concept that can be used in the development of vaccines based on HAdV vectors. Our data suggests that different immunomodulatory potentials can be observed in the conserved regions, present in the hexon and penton proteins, from different species. Using this modular approach, we developed a HAdV-5 based vaccine strategy for SARS-CoV-2, constructed through the display of SARS-CoV-2 epitopes indicated by our prediction analysis as immunologically relevant. The sequences of the HAdV vector major capsid proteins were also edited to enhance the IFN-gamma induction and antigen presenting cells activation. This is the first study proposing a modular HAdV platform developed to aid the design of new vaccines by inducing an immune response more suited for the epitopes in display.
Assuntos
Proteínas do Capsídeo/química , Biologia Computacional/métodos , Epitopos de Linfócito B/imunologia , Vacinas Virais/imunologia , Apresentação de Antígeno , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/imunologia , Simulação por Computador , Dependovirus/imunologia , Desenho de Fármacos , Epitopos de Linfócito B/genética , Humanos , Imunidade Humoral , Interferon gama/metabolismo , SARS-CoV-2/efeitos dos fármacos , SARS-CoV-2/genética , SARS-CoV-2/imunologia , Vacinas Virais/genéticaRESUMO
Since the beginning of COVID-19 pandemic studies on viral shedding have reported that this virus is excreted in feces in most patients. High viral loads are found at the sewage pipeline or at the entrance of wastewater treatment plants from cities where the number of COVID-19 cases are significant. In Quito (Ecuador) as in many other cities worldwide, wastewater is directly discharged into natural waters. The aim of this study was to evaluate SARS-CoV-2 presence in urban streams from a low sanitation context. Three river locations along the urban rivers of Quito were sampled on the 5th of June during a peak of COVID-19 cases. River samples were evaluated for water quality parameters and afterwards, concentrated for viral analysis using skimmed milk flocculation method. The viral concentrates were quantified for SARS-CoV-2 (N1 and N2 target regions) and Human Adenovirus as a human viral indicator. The results showed that SARS-CoV-2 was detected for both target regions in all samples analyzed in a range of 2,91E+05 to 3,19E+06 GC/L for N1 and from 2,07E+05 to 2,22E+06 GC/L for N2. The high values detected in natural waters from a low sanitation region have several implications in health and ecology that should be further assessed.
Assuntos
Infecções por Coronavirus , Pandemias , Pneumonia Viral , Rios , Saneamento , Betacoronavirus , COVID-19 , Cidades , Equador , Humanos , SARS-CoV-2RESUMO
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
Background: The Brazilian domestic canine population are the second largest in the world and their feeding means 0.4%of the Brazilian gross domestic product. For maintaining the quality of the food, the companies use worldwide standardsfor technical prevention and control of contaminants and biological conservation. The packaging is part of this process,since it provides a barrier between food and environment. However, in Brazil, packagings are often opened in retail storesfor bulk marketing. The objectives of this work were to develop a methodology to detect viruses in foods and to analyzethe bacterial and fungal contamination in puppies food sold in bulk in Ivoti and Estância Velha, cities in Southern Brazil.Materials, Methods & Results: Twenty samples collected between September and October 2016 were analyzed for mostprobable number of coliforms, Salmonella sp., mesophilic aerobic bacteria and yeast/mold following the regulation of theBrazilian Ministry of Agriculture, Livestock and Food Supply guidelines. They were also tested for Human MastadenovirusC (HAdV), Canine Mastadenovirus A (CAdV), and Carnivore Protoparvovirus 1 (CPV) genomes. Viral analysis wereperformed by polymerase chain reaction (PCR) detection. During the collection of the samples hygienic-sanitary conditions, storage of feeds, animals access, dog grooming, and veterinary care were considered to evaluate the conditions ofeach store. A pilot study was carried out using one food sample marketed in bulk and one sample from the original package(closed package) and testing them for bacterial and fungal contamination for standardizing viral detection. Ten grams offood from the original package were mixed with 90 mL of Eagles Minimal Essential Medium (E-MEM) in 100 mL sterilebottles. These bottles were kept in room temperature and shaken for 60 min. Subsequently, aliquots were obtained by sequentially diluting the sample (10-2 to 10-4)...(AU)
Assuntos
Animais , Cães , Adenovírus Humanos/isolamento & purificação , Ração Animal/microbiologia , Salmonella/isolamento & purificação , Aspergillus/isolamento & purificação , Penicillium/isolamento & purificação , Contaminação de Alimentos/análise , Fungos , Bactérias Aeróbias , Reação em Cadeia da PolimeraseRESUMO
Human mastadenovirus (HAdV) genus is related to several diseases, among them upper and lower respiratory tract illness. HAdV species B, C, D, and E are mainly associated with respiratory infections. The goal of this work was to identify the HAdV species associated with respiratory infections in hospitalized patients from southern Brazil. Samples were collected from 1996 to 2004 and 2011 to 2017. During this period, 28,524 samples were collected, and 9983 were positive for respiratory viruses, being 435 for HAdV. From these 435 samples, 57 were selected for characterization of HAdV species. For screening the presence of HAdV, a partial sequence of the DNA polymerase gene (DNApol gene) was amplified by nested PCR. Partial nucleotide sequencing was performed in positive samples, and HAdV (DNApol gene) was detected in 53 samples: species B (28;49.1%), C (16;8.0%), D (2; 3.5%), E (5; 8.7%), and untyped (2; 3.5%). Specie D was found only in 2017 and specie E in 2011 and 2012. The age of the patients ranged from < 1 to 81 years old, and 62.3%were male. No relationship between gender orage and identified HAdV species were observed. In addition, in the period of 20132017, 18 samples from patients who died were analyzed: 11 were related to species B, 4 to C, and 2 to D and 1 remained untyped. Circulation of HAdV species D and Evaried over the years, but species B and C were present throughout the evaluated period. In addition, respiratory infections by HAdVaffect elderly and children mainly. (AU)
Assuntos
Humanos , Masculino , Feminino , Criança , Idoso , Idoso de 80 Anos ou mais , Sistema Respiratório , Infecções Respiratórias/virologia , Mastadenovirus/patogenicidade , Ácidos Nucleicos , MorbidadeRESUMO
Anthropogenic contamination of beaches in the south of Brazil was assessed by detection of Escherichia coli, human mastadenovirus species C (HAdV-C) and F (HAdV-F) and hepatitis E virus (HEV). Sampling was carried out in October (2016), and in January, April and July (2017). Water, sediment, sea surface microlayer (SML), bivalves, and air sentinel samples were evaluated. Quantitative microbiological risk assessment (QMRA) was used to estimate the probability of swimmer infection. HAdV-C was present in 26% of the samples, for both qPCR and viral isolation. The highest rates of detection in genomic copies (GC) were in water (2.42E+10 GC/L), SML (2.08E+10 GC/L), sediment (3.82E+08 GC/g) and bivalves (3.91E+07 GC/g). QMRA estimated daily and annual risks with a maximum value (9.99E-01) in almost all of the samples. Viable HAdV-C was often detected in the SML, pointing that this is a source of infection for people bathing in these waters.
Assuntos
Adenovírus Humanos/isolamento & purificação , Bivalves/virologia , Sedimentos Geológicos/virologia , Água do Mar/virologia , Adenovírus Humanos/genética , Animais , Praias , Brasil , Monitoramento Ambiental , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Vírus da Hepatite E/genética , Vírus da Hepatite E/isolamento & purificação , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Medição de Risco/métodos , Natação , Microbiologia da ÁguaRESUMO
Human mastadenovirus (HAdV) genus is related to several diseases, among them upper and lower respiratory tract illness. HAdV species B, C, D, and E are mainly associated with respiratory infections. The goal of this work was to identify the HAdV species associated with respiratory infections in hospitalized patients from southern Brazil. Samples were collected from 1996 to 2004 and 2011 to 2017. During this period, 28,524 samples were collected, and 9983 were positive for respiratory viruses, being 435 for HAdV. From these 435 samples, 57 were selected for characterization of HAdV species. For screening the presence of HAdV, a partial sequence of the DNA polymerase gene (DNApol gene) was amplified by nested PCR. Partial nucleotide sequencing was performed in positive samples, and HAdV (DNApol gene) was detected in 53 samples: species B (28; 49.1%), C (16; 28.0%), D (2; 3.5%), E (5; 8.7%), and untyped (2; 3.5%). Specie D was found only in 2017 and specie E in 2011 and 2012. The age of the patients ranged from < 1 to 81 years old, and 62.3% were male. No relationship between gender or age and identified HAdV species were observed. In addition, in the period of 2013-2017, 18 samples from patients who died were analyzed: 11 were related to species B, 4 to C, and 2 to D and 1 remained untyped. Circulation of HAdV species D and E varied over the years, but species B and C were present throughout the evaluated period. In addition, respiratory infections by HAdV affect elderly and children mainly.
Assuntos
Infecções por Adenoviridae/virologia , Mastadenovirus/isolamento & purificação , Infecções Respiratórias/virologia , Infecções por Adenoviridae/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Mastadenovirus/classificação , Mastadenovirus/genética , Pessoa de Meia-Idade , Filogenia , Infecções Respiratórias/epidemiologia , Adulto JovemRESUMO
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
Human adenovirus (HAdV-2) is considered a common agent of respiratory tract infection in the human, especially in children. Virus infection is believed to modify host cell expression necessary for its replication and therefore cell proteome can reflect the changes of specific cellular pathways during infection. This study aims to identify differentially expressed proteins of A549 cells in response to HAdV-2 infection using a label-free liquid chromatography-high-resolution tandem mass spectrometry strategy (LC-MS/MS) at 24 and 48 hpi. A total of 248 and 216 proteins were deregulated by 1.35-fold at 24 and 48 hpi, respectively. Among them, 155 were upregulated at 24 hpi and 86 at 48 hpi, whereas 93 and 130 were downregulated at 24 and 48 hpi, respectively. The identified proteins were involved in different pathways as energy, transcription, protein synthesis, cytoskeleton, rescue and defense, cell cycle, DNA processing, transportation, and metabolism. Glycolytic pathway and histone deregulated proteins were further confirmed by chemical testing and immunofluorescence, respectively. The results suggest that the identified proteins influenced HAdV-2 infection in the context of viral replication and propagation. This study complement proteomic data obtained from previous studies and reinforce the understanding of the relationship between HAdV and host cell.
Assuntos
Adenovírus Humanos/genética , Interações Hospedeiro-Patógeno , Proteômica , Células A549 , Adenovírus Humanos/fisiologia , Cromatografia Líquida , Regulação para Baixo , Glucose/metabolismo , Humanos , Proteoma/análise , Espectrometria de Massas em Tandem , Replicação ViralRESUMO
Background: The Brazilian domestic canine population are the second largest in the world and their feeding means 0.4%of the Brazilian gross domestic product. For maintaining the quality of the food, the companies use worldwide standardsfor technical prevention and control of contaminants and biological conservation. The packaging is part of this process,since it provides a barrier between food and environment. However, in Brazil, packagings are often opened in retail storesfor bulk marketing. The objectives of this work were to develop a methodology to detect viruses in foods and to analyzethe bacterial and fungal contamination in puppies food sold in bulk in Ivoti and Estância Velha, cities in Southern Brazil.Materials, Methods & Results: Twenty samples collected between September and October 2016 were analyzed for mostprobable number of coliforms, Salmonella sp., mesophilic aerobic bacteria and yeast/mold following the regulation of theBrazilian Ministry of Agriculture, Livestock and Food Supply guidelines. They were also tested for Human MastadenovirusC (HAdV), Canine Mastadenovirus A (CAdV), and Carnivore Protoparvovirus 1 (CPV) genomes. Viral analysis wereperformed by polymerase chain reaction (PCR) detection. During the collection of the samples hygienic-sanitary conditions, storage of feeds, animals access, dog grooming, and veterinary care were considered to evaluate the conditions ofeach store. A pilot study was carried out using one food sample marketed in bulk and one sample from the original package(closed package) and testing them for bacterial and fungal contamination for standardizing viral detection. Ten grams offood from the original package were mixed with 90 mL of Eagles Minimal Essential Medium (E-MEM) in 100 mL sterilebottles. These bottles were kept in room temperature and shaken for 60 min. Subsequently, aliquots were obtained by sequentially diluting the sample (10-2 to 10-4)...
Assuntos
Animais , Cães , Adenovírus Humanos/isolamento & purificação , Aspergillus/isolamento & purificação , Penicillium/isolamento & purificação , Ração Animal/microbiologia , Salmonella/isolamento & purificação , Bactérias Aeróbias , Contaminação de Alimentos/análise , Fungos , Reação em Cadeia da PolimeraseRESUMO
A novel human adenovirus was isolated from a pediatric case of acute respiratory disease in Panama City, Panama in 2011. The clinical isolate was initially identified as an intertypic recombinant based on hexon and fiber gene sequencing. Based on the analysis of its complete genome sequence, the novel complex recombinant Human mastadenovirus D (HAdV-D) strain was classified into a new HAdV type: HAdV-84, and it was designated Adenovirus D human/PAN/P309886/2011/84[P43H17F84]. HAdV-D types possess usually an ocular or gastrointestinal tropism, and respiratory association is scarcely reported. The virus has a novel fiber type, most closely related to, but still clearly distant from that of HAdV-36. The predicted fiber is hypothesised to bind sialic acid with lower affinity compared to HAdV-37. Bioinformatic analysis of the complete genomic sequence of HAdV-84 revealed multiple homologous recombination events and provided deeper insight into HAdV evolution.
Assuntos
Adenovírus Humanos/genética , Proteínas do Capsídeo/genética , Genoma Viral , Genótipo , Análise de Sequência de DNA , Infecções por Adenoviridae/virologia , Adenovírus Humanos/isolamento & purificação , Proteínas do Capsídeo/metabolismo , Humanos , Panamá , Filogenia , Ligação Proteica , Recombinação Genética , Homologia de Sequência , Ácidos Siálicos/metabolismoRESUMO
Human adenovirus (HAdV) is resistant to environment and can be used as a marker to detect fecal contamination. Considering the importance of freshwater snails in the aquatic environment, their use as concentrators for HAdV is a complementary tool for viral analysis of water. The goal of the study was to detect HAdV in snails and surface water collected from wetlands of the Sinos River (Rio Grande do Sul, Brazil) basin and to compare rates and viral loads found in both samples. HAdV was detected through real-time PCR. Total and fecal coliforms were detected by Colilert® kit, and viral infectivity of positive samples of the DNA genome was performed in A549 human cell line. All wetlands presented bacterial and viral contamination, but no viral particle was considered viable. The wetland that showed lower fecal coliform mean was Campo Bom, and São Leopoldo (both cities in Rio Grande do Sul) was representative of the highest mean. HAdV was detected in water samples (53%), gastropods' hemolymph (31%) and tissues (16%). Wetlands proved to be environments already altered by human action. Water samples exhibited a higher frequency of HAdV detection; however, in some instances, the target viral genomes were only found in gastropod biological samples. This was a pioneer study in the use of freshwater snails for human enteric viral assessment thus demonstrating that the human organism can retain fecal contamination, complementing and assisting in microbiological water analyzes.
Assuntos
Adenovírus Humanos/crescimento & desenvolvimento , Monitoramento Ambiental , Água Doce/virologia , Caramujos/virologia , Animais , Brasil , Cidades , Fezes , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Rios , Microbiologia da Água , Poluição da Água/análise , Poluição da Água/estatística & dados numéricosRESUMO
Climate variables may interfere with the environmental persistence and spread of pathogenic microorganisms. This study aimed to investigate the occurrence of human adenovirus (HAdV) and total and thermotolerant coliforms in treated and untreated water and report gastroenteritis cases in seven cities located in the hydrographic basin of the Sinos River (HBSR), Southern Brazil. The data on water quality from samples collected at catchment areas of HBSR from March to December 2011 were compared with precipitation records, virus detection rates and viral loads, and information on enteric diseases among residents of the region. There was a marked increase in precipitation intensity in April, July, and August and a decrease in May and November. The number of HAdV genome copies (gc) in untreated water ranged from 2.1×10(8) gc/L in June to 7.8×10(1) gc/L in December, and in treated water, from 6.3×10(4) gc/L in September to 4.1×10(1) gc/L in November. The most probable number (MPN) of total coliforms ranged from 5×10(1) MPN/100 mL in December to 2.4×10(5) MPN/100 mL in July, and thermotolerant coliforms ranged from 1×10(1) MPN/100 mL in August to 6.9×10(4) MPN/100 mL in July. A total of 79 hospital admissions due to gastroenteritis were registered in the cities studied. The results for coliforms in untreated water demonstrate deficits in sanitation and wastewater treatment. These findings also indicate a possible relationship between the occurrence of rainfalls after dry periods and an increase in the number of gastroenteritis cases and in HAdV load quantified in surface water collected for conventional potabilization.
Assuntos
Adenovírus Humanos , Gastroenterite/epidemiologia , Microbiologia da Água , Brasil/epidemiologia , Cidades/epidemiologia , Monitoramento Ambiental/métodos , Humanos , Chuva/virologia , Rios/virologia , Purificação da Água , Qualidade da ÁguaRESUMO
Human adenoviruses (HAdV), members of the Adenoviridae family, are excreted through the fecal route and may be present in the feces of humans consuming contaminated food or water. The presence of HAdV from different serotypes in the feces of healthy individuals was already reported using conventional polymerase chain reaction; however, real-time PCR (qPCR) may reveal not only the rates of detection as well as demonstrate the viral loads excreted by healthy persons. Aiming to identify and characterize the presence of adenoviruses in stool samples, 147 fecal samples from patients with no records of diarrhea were analyzed (74 from winter season and 73 from summer) by Real-Time PCR (qPCR) assay and conventional PCR. HAdV genome was present in 43.8% (32/73) of stools samples collected during summer season and 21.6% (16/74) during winter. The rate of detection of genomic copies (gc) ranged from 4.04×10(2) to 6.72×10(5)gc/g of feces among the 147 samples analyzed, of which the ranged of genomic copies of DNA HAdV was major in summer. All samples were negative when tested for rotaviruses (RV) and noroviruses (NoV) by PCR conventional and qPCR respectively. HAdV is excreted constantly by infected individuals in the absence of clinical signs and the occurrence may vary seasonally.