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1.
Viruses ; 13(2)2021 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-33525386

RESUMO

Parvoviruses are small single-stranded DNA viruses that can infect both vertebrates and invertebrates. We report here the full characterization of novel viruses we identified in ducks, including two viral species within the subfamily Hamaparvovirinae (duck-associated chapparvovirus, DAC) and a novel species within the subfamily Densovirinae (duck-associated ambidensovirus, DAAD). Overall, 5.7% and 21.1% of the 123 screened ducks (American black ducks, mallards, northern pintail) were positive for DAC and DAAD, respectively, and both viruses were more frequently detected in autumn than in winter. Genome organization and predicted transcription profiles of DAC and DAAD were similar to viruses of the genera Chaphamaparvovirus and Protoambidensovirus, respectively. Their association to these genera was also demonstrated by subfamily-wide phylogenetic and distance analyses of non-structural protein NS1 sequences. While DACs were included in a highly supported clade of avian viruses, no definitive conclusions could be drawn about the host type of DAAD because it was phylogenetically close to viruses found in vertebrates and invertebrates and analyses of codon usage bias and nucleotide frequencies of viruses within the family Parvoviridae showed no clear host-based viral segregation. This study highlights the high parvoviral diversity in the avian reservoir with many avian-associated parvoviruses likely yet to be discovered.


Assuntos
Patos/virologia , Infecções por Parvoviridae/veterinária , Parvoviridae/genética , Animais , Animais Selvagens/virologia , Uso do Códon , DNA Viral/genética , Patos/classificação , Genoma Viral/genética , Especificidade de Hospedeiro , Parvoviridae/classificação , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Filogenia , Estações do Ano , Proteínas não Estruturais Virais/genética
2.
Transbound Emerg Dis ; 68(2): 802-812, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32688446

RESUMO

Canine bufavirus (CBuV) is a protoparvovirus, genetically related to human and non-human primate bufaviruses and distantly related to canine parvovirus type 2 (CPV-2). CBuV was initially identified from young dogs with respiratory signs but subsequent studies revealed that this virus is also a common component of the canine enteric virome. In this survey, by assessing archival and recent collections of dogs faecal samples, CBuV DNA was detected with a higher prevalence rate (8.8%) in animals with enteritis than in control animals (5.0%), although this difference was not statistically significant. The rate of co-infections with other enteric viruses in diarrhoeic dogs was high (84.6%), mostly in association with canine parvovirus CPV-2 (90.1%). The complete ORF2 gene was determined in five samples, and the nearly full-length genome was reconstructed for three strains, 62/2017/ITA, 9AS/2005/ITA and 35/2018/ITA. Upon sequence comparison, the viruses appeared highly conserved in the NS1 (97.2%-97.9% nt and 97.5%-98.1% aa identities). In the complete VP2 coding region, three strains were similar to the prototype viruses (99.7-99.8 nt and 99.6%-99.8% aa) whilst strains 9AS/2005/ITA and 35/2016/ITA were distantly related (87.6%-89.3% nt and 93.9%-95.1% aa identities). Interestingly, genetic diversification occurred downstream conserved regions such as the VP1/VP2 splicing signals and/or the G-rich motif in the N terminus of the VP2, suggesting a potential recombination nature. Upon phylogenetic analysis, the two divergent CBuV strains formed a distinct cluster/genotype.


Assuntos
Doenças do Cão/virologia , Heterogeneidade Genética , Infecções por Parvoviridae/veterinária , Parvoviridae/genética , Animais , Doenças do Cão/epidemiologia , Cães , Fezes/virologia , Genótipo , Parvoviridae/classificação , Infecções por Parvoviridae/virologia , Filogenia
3.
PLoS One ; 15(12): e0244056, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33332429

RESUMO

Viral infections were investigated in American black bears (Ursus americanus) from Nevada and northern California with and without idiopathic encephalitis. Metagenomics analyses of tissue pools revealed novel viruses in the genera Circoviridae, Parvoviridae, Anelloviridae, Polyomaviridae, and Papillomaviridae. The circovirus and parvovirus were of particular interest due to their potential importance as pathogens. We characterized the genomes of these viruses and subsequently screened bears by PCR to determine their prevalence. The circovirus (Ursus americanus circovirus, UaCV) was detected at a high prevalence (10/16, 67%), and the chaphamaparvovirus (Ursus americanus parvovirus, UaPV) was found in a single bear. We showed that UaCV is present in liver, spleen/lymph node, and brain tissue of selected cases by in situ hybridization (ISH) and PCR. Infections were detected in cases of idiopathic encephalitis and in cases without inflammatory brain lesions. Infection status was not clearly correlated with disease, and the significance of these infections remains unclear. Given the known pathogenicity of a closely related mammalian circovirus, and the complex manifestations of circovirus-associated diseases, we suggest that UaCV warrants further study as a possible cause or contributor to disease in American black bears.


Assuntos
Doenças dos Animais/virologia , Circoviridae/patogenicidade , Encefalite Viral/virologia , Parvoviridae/patogenicidade , Ursidae/virologia , Doenças dos Animais/epidemiologia , Animais , Encéfalo/virologia , Circoviridae/genética , Circoviridae/isolamento & purificação , Código de Barras de DNA Taxonômico , Encefalite Viral/epidemiologia , Fígado/virologia , Metagenoma , Parvoviridae/genética , Parvoviridae/isolamento & purificação , Baço/virologia , Estados Unidos
5.
Viruses ; 12(10)2020 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-32992674

RESUMO

Viral pathogens are being increasingly described in association with mass morbidity and mortality events in reptiles. However, our knowledge of reptile viruses remains limited. Herein, we describe the meta-transcriptomic investigation of a mass morbidity and mortality event in a colony of central bearded dragons (Pogona vitticeps) in 2014. Severe, extensive proliferation of the respiratory epithelium was consistently found in affected dragons. Similar proliferative lung lesions were identified in bearded dragons from the same colony in 2020 in association with increased intermittent mortality. Total RNA sequencing identified two divergent DNA viruses: a reptile-infecting circovirus, denoted bearded dragon circovirus (BDCV), and the first exogeneous reptilian chaphamaparvovirus-bearded dragon chaphamaparvovirus (BDchPV). Phylogenetic analysis revealed that BDCV was most closely related to bat-associated circoviruses, exhibiting 70% amino acid sequence identity in the Replicase (Rep) protein. In contrast, in the nonstructural (NS) protein, the newly discovered BDchPV showed approximately 31%-35% identity to parvoviruses obtained from tilapia fish and crocodiles in China. Subsequent specific PCR assays revealed BDCV and BDchPV in both diseased and apparently normal captive reptiles, although only BDCV was found in those animals with proliferative pulmonary lesions and respiratory disease. This study expands our understanding of viral diversity in captive reptiles.


Assuntos
Circovirus/isolamento & purificação , Parvoviridae/isolamento & purificação , Répteis/virologia , Infecções Respiratórias/veterinária , Animais , China/epidemiologia , Circovirus/classificação , Circovirus/genética , Circovirus/patogenicidade , Genoma Viral/genética , Lagartos/virologia , Pulmão/patologia , Parvoviridae/classificação , Parvoviridae/genética , Parvoviridae/patogenicidade , Filogenia , Prevalência , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/patologia , Infecções Respiratórias/virologia , Proteínas Virais/genética
6.
PLoS Pathog ; 16(8): e1008718, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32797103

RESUMO

APOBEC3 enzymes are innate immune effectors that introduce mutations into viral genomes. These enzymes are cytidine deaminases which transform cytosine into uracil. They preferentially mutate cytidine preceded by thymidine making the 5'TC motif their favored target. Viruses have evolved different strategies to evade APOBEC3 restriction. Certain viruses actively encode viral proteins antagonizing the APOBEC3s, others passively face the APOBEC3 selection pressure thanks to a depleted genome for APOBEC3-targeted motifs. Hence, the APOBEC3s left on the genome of certain viruses an evolutionary footprint. The aim of our study is the identification of these viruses having a genome shaped by the APOBEC3s. We analyzed the genome of 33,400 human viruses for the depletion of APOBEC3-favored motifs. We demonstrate that the APOBEC3 selection pressure impacts at least 22% of all currently annotated human viral species. The papillomaviridae and polyomaviridae are the most intensively footprinted families; evidencing a selection pressure acting genome-wide and on both strands. Members of the parvoviridae family are differentially targeted in term of both magnitude and localization of the footprint. Interestingly, a massive APOBEC3 footprint is present on both strands of the B19 erythroparvovirus; making this viral genome one of the most cleaned sequences for APOBEC3-favored motifs. We also identified the endemic coronaviridae as significantly footprinted. Interestingly, no such footprint has been detected on the zoonotic MERS-CoV, SARS-CoV-1 and SARS-CoV-2 coronaviruses. In addition to viruses that are footprinted genome-wide, certain viruses are footprinted only on very short sections of their genome. That is the case for the gamma-herpesviridae and adenoviridae where the footprint is localized on the lytic origins of replication. A mild footprint can also be detected on the negative strand of the reverse transcribing HIV-1, HIV-2, HTLV-1 and HBV viruses. Together, our data illustrate the extent of the APOBEC3 selection pressure on the human viruses and identify new putatively APOBEC3-targeted viruses.


Assuntos
Citidina Desaminase/metabolismo , Genoma Viral/genética , Interações Hospedeiro-Patógeno/genética , Seleção Genética/genética , Replicação Viral/genética , Coronaviridae/genética , Humanos , Imunidade Inata/imunologia , Papillomaviridae/genética , Parvoviridae/genética , Polyomaviridae/genética , Proteínas Virais/genética
7.
Hepatology ; 72(6): 2182-2196, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32654263

RESUMO

BACKGROUND AND AIMS: Alcoholic hepatitis (AH) is a severe manifestation of alcohol-associated liver disease (ALD) with high mortality. Although gut bacteria and fungi modulate disease severity, little is known about the effects of the viral microbiome (virome) in patients with ALD. APPROACH AND RESULTS: We extracted virus-like particles from 89 patients with AH who were enrolled in a multicenter observational study, 36 with alcohol use disorder (AUD), and 17 persons without AUD (controls). Virus-like particles from fecal samples were fractionated using differential filtration techniques, and metagenomic sequencing was performed to characterize intestinal viromes. We observed an increased viral diversity in fecal samples from patients with ALD, with the most significant changes in samples from patients with AH. Escherichia-, Enterobacteria-, and Enterococcus phages were over-represented in fecal samples from patients with AH, along with significant increases in mammalian viruses such as Parvoviridae and Herpesviridae. Antibiotic treatment was associated with higher viral diversity. Specific viral taxa, such as Staphylococcus phages and Herpesviridae, were associated with increased disease severity, indicated by a higher median Model for End-Stage Liver Disease score, and associated with increased 90-day mortality. CONCLUSIONS: In conclusion, intestinal viral taxa are altered in fecal samples from patients with AH and associated with disease severity and mortality. Our study describes an intestinal virome signature associated with AH.


Assuntos
Doença Hepática Terminal/virologia , Hepatite Alcoólica/virologia , Mucosa Intestinal/virologia , Cirrose Hepática/virologia , Viroma/genética , Adulto , Idoso , Animais , Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Estudos de Casos e Controles , DNA Viral/isolamento & purificação , Doença Hepática Terminal/diagnóstico , Doença Hepática Terminal/mortalidade , Doença Hepática Terminal/terapia , Fezes/virologia , Feminino , Hepatite Alcoólica/diagnóstico , Hepatite Alcoólica/mortalidade , Hepatite Alcoólica/terapia , Herpesviridae/genética , Herpesviridae/isolamento & purificação , Humanos , Fígado/patologia , Cirrose Hepática/diagnóstico , Cirrose Hepática/mortalidade , Cirrose Hepática/terapia , Masculino , Metagenômica , Pessoa de Meia-Idade , Parvoviridae/genética , Parvoviridae/isolamento & purificação , RNA Viral/isolamento & purificação , Índice de Gravidade de Doença , Taxa de Sobrevida
8.
Infect Genet Evol ; 85: 104421, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32580027

RESUMO

Crab-eating (Cerdocyon thous) and Pampas foxes (Lycalopex gymnocercus) are wild canids distributed in South America. Domestic dogs (Canis lupus familiaris) and wild canids may share viral pathogens, including rabies virus (RABV), canine distemper virus (CDV), and canine parvovirus 2 (CPV-2). To characterize the virome of these wild canid species, the present work evaluated the spleen and mesenteric lymph node virome of 17 crab-eating and five Pampas foxes using high-throughput sequencing (HTS). Organ samples were pooled and sequenced using an Illumina MiSeq platform. Additional PCR analyses were performed to identify the frequencies and host origin for each virus detected by HTS. Sequences more closely related to the Paramyxoviridae, Parvoviridae and Anelloviridae families were detected, as well as circular Rep-encoding single-stranded (CRESS) DNA viruses. CDV was found only in crab-eating foxes, whereas CPV-2 was found in both canid species; both viruses were closely related to sequences reported in domestic dogs from southern Brazil. Moreover, the present work reported the detection of canine bocavirus (CBoV) strains that were genetically divergent from CBoV-1 and 2 lineages. Finally, we also characterized CRESS DNA viruses and anelloviruses with marked diversity. The results of this study contribute to the body of knowledge regarding wild canid viruses that can potentially be shared with domestic canids or other species.


Assuntos
Cães/virologia , Raposas/virologia , Viroma , Vírus/classificação , Vírus/genética , Anelloviridae/classificação , Anelloviridae/genética , Animais , Bocavirus/classificação , Bocavirus/genética , Brasil , Vírus de DNA/classificação , Vírus de DNA/genética , DNA Viral , Vírus da Cinomose Canina/classificação , Vírus da Cinomose Canina/genética , Sequenciamento de Nucleotídeos em Larga Escala , Linfonodos/virologia , Metagenômica , Paramyxoviridae/classificação , Paramyxoviridae/genética , Parvoviridae/classificação , Parvoviridae/genética , Parvovirus Canino/classificação , Parvovirus Canino/genética , Filogenia , RNA Viral , Baço/virologia , Uruguai , Viroses/veterinária , Viroses/virologia , Vírus/isolamento & purificação
9.
Arch Virol ; 165(9): 2133-2146, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32533329

RESUMO

Parvoviridae, a diverse family of small single-stranded DNA viruses was established in 1975. It was divided into two subfamilies, Parvovirinae and Densovirinae, in 1993 to accommodate parvoviruses that infect vertebrate and invertebrate animals, respectively. This relatively straightforward segregation, using host association as the prime criterion for subfamily-level classification, has recently been challenged by the discovery of divergent, vertebrate-infecting parvoviruses, dubbed "chapparvoviruses", which have proven to be more closely related to viruses in certain Densovirinae genera than to members of the Parvovirinae. Viruses belonging to these genera, namely Brevi-, Hepan- and Penstyldensovirus, are responsible for the unmatched heterogeneity of the subfamily Densovirinae when compared to the Parvovirinae in matters of genome organization, protein sequence homology, and phylogeny. Another genus of Densovirinae, Ambidensovirus, has challenged traditional parvovirus classification, as it includes all newly discovered densoviruses with an ambisense genome organization, which introduces genus-level paraphyly. Lastly, current taxon definition and virus inclusion criteria have significantly limited the classification of certain long-discovered parvoviruses and impedes the classification of some potential family members discovered using high-throughput sequencing methods. Here, we present a new and updated system for parvovirus classification, which includes the introduction of a third subfamily, Hamaparvovirinae, resolves the paraphyly within genus Ambidensovirus, and introduces new genera and species into the subfamily Parvovirinae. These proposals were accepted by the ICTV in 2020 March.


Assuntos
Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/virologia , Parvoviridae/classificação , Parvoviridae/fisiologia , Filogenia , Animais , Especificidade de Hospedeiro , Humanos , Parvoviridae/genética , Parvoviridae/isolamento & purificação , Proteínas Virais/genética
11.
J Virol ; 94(18)2020 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-32581107

RESUMO

Wild birds are major natural reservoirs and potential dispersers of a variety of infectious diseases. As such, it is important to determine the diversity of viruses they carry and use this information to help understand the potential risks of spillover to humans, domestic animals, and other wildlife. We investigated the potential viral causes of paresis in long-standing, but undiagnosed, disease syndromes in wild Australian birds. RNA from diseased birds was extracted and pooled based on tissue type, host species, and clinical manifestation for metagenomic sequencing. Using a bulk and unbiased metatranscriptomic approach, combined with clinical investigation and histopathology, we identified a number of novel viruses from the families Astroviridae, Adenoviridae, Picornaviridae, Polyomaviridae, Paramyxoviridae, Parvoviridae, and Circoviridae in common urban wild birds, including Australian magpies, magpie larks, pied currawongs, Australian ravens, and rainbow lorikeets. In each case, the presence of the virus was confirmed by reverse transcription (RT)-PCR. These data revealed a number of candidate viral pathogens that may contribute to coronary, skeletal muscle, vascular, and neuropathology in birds of the Corvidae and Artamidae families and neuropathology in members of the Psittaculidae The existence of such a diverse virome in urban avian species highlights the importance and challenges in elucidating the etiology and ecology of wildlife pathogens in urban environments. This information will be increasingly important for managing disease risks and conducting surveillance for potential viral threats to wildlife, livestock, and human health.IMPORTANCE Wildlife naturally harbor a diverse array of infectious microorganisms and can be a source of novel diseases in domestic animals and human populations. Using unbiased RNA sequencing, we identified highly diverse viruses in native birds from Australian urban environments presenting with paresis. This research included the clinical investigation and description of poorly understood recurring syndromes of unknown etiology: clenched claw syndrome and black and white bird disease. As well as identifying a range of potentially disease-causing viral pathogens, this study describes methods that can effectively and efficiently characterize emergent disease syndromes in free-ranging wildlife and promotes further surveillance for specific pathogens of potential conservation and zoonotic concern.


Assuntos
Animais Selvagens/virologia , Doenças das Aves/epidemiologia , Aves/virologia , Infecções por Vírus de DNA/veterinária , Metagenoma , Infecções por Vírus de RNA/veterinária , Transcriptoma , Adenoviridae/classificação , Adenoviridae/genética , Adenoviridae/isolamento & purificação , Animais , Astroviridae/classificação , Astroviridae/genética , Astroviridae/isolamento & purificação , Austrália/epidemiologia , Doenças das Aves/virologia , Circoviridae/classificação , Circoviridae/genética , Circoviridae/isolamento & purificação , Cidades , Infecções por Vírus de DNA/epidemiologia , Infecções por Vírus de DNA/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Paramyxoviridae/classificação , Paramyxoviridae/genética , Paramyxoviridae/isolamento & purificação , Parvoviridae/classificação , Parvoviridae/genética , Parvoviridae/isolamento & purificação , Filogenia , Picornaviridae/classificação , Picornaviridae/genética , Picornaviridae/isolamento & purificação , Polyomaviridae/classificação , Polyomaviridae/genética , Polyomaviridae/isolamento & purificação , Infecções por Vírus de RNA/epidemiologia , Infecções por Vírus de RNA/virologia
12.
Infect Genet Evol ; 81: 104203, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32035977

RESUMO

Brazil is a major exporter of pork meat worldwide. Swine liver is a common ingredient in food consumed by humans, thus emphasizing the importance of evaluating the presence of associated pathogens in swine liver. To obtain knowledge, this study aimed to provide insights into the viral communities of livers collected from slaughtered pigs from southern Brazil. The 46 livers were processed and submitted for high-throughput sequencing (HTS). The sequences were most closely related to Anelloviridae, Circoviridae and Parvoviridae families. The present work also describes the first Brazilian PCV1 and the first PPV6 and PPV7 from South America. Virus frequencies revelead 63% of samples positive for TTSuV1, 71% for TTSuVk2, 10.8% for PCV, 13% for PPV and 6% for PBov. This report addresses the diversity of the liver virome of healthy pigs and expands the number of viruses detected, further characterizing their genomes to assist future studies.


Assuntos
Vírus de DNA/genética , DNA de Cadeia Simples/genética , Genoma Viral/genética , Fígado/virologia , Suínos/virologia , Viroma/genética , Anelloviridae/genética , Animais , Brasil , Circoviridae/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Parvoviridae/genética , Doenças dos Suínos/virologia
13.
J Gen Virol ; 101(2): 226-239, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31855134

RESUMO

Diaphorina citri densovirus (DcDV) is an ambisense densovirus with a 5071 nt genome. Phylogenetic analysis places DcDV in an intermediate position between those in the Ambidensovirus and Iteradensovirus genera, a finding that is consistent with the observation that DcDV possesses an Iteradensoviris-like non-structural (NS) protein-gene cassette, but a capsid-protein (VP) gene cassette resembling those of other ambisense densoviruses. DcDV is maternally transmitted to 100 % of the progeny of infected female Diaphorina citri, and the progeny of infected females carry DcDV as a persistent infection without outward phenotypic effects. We were unable to infect naïve individuals by oral inoculation, however low levels of transient viral replication are detected following intrathoracic injection of DcDV virions into uninfected D. citri insects. Transcript mapping indicates that DcDV produces one transcript each from the NS and VP gene cassettes and that these transcripts are polyadenylated at internal sites to produce a ~2.2 kb transcript encoding the NS proteins and a ~2.4 kb transcript encoding the VP proteins. Additionally, we found that transcriptional readthrough leads to the production of longer non-canonical transcripts from both genomic strands.


Assuntos
Densovirus , Genoma Viral , Hemípteros/virologia , Viroses/transmissão , Animais , Proteínas do Capsídeo/genética , Classificação , Vírus de DNA/genética , Densovirus/classificação , Densovirus/genética , Densovirus/isolamento & purificação , Genes Virais , Transmissão Vertical de Doenças Infecciosas , Vírus de Insetos/classificação , Parvoviridae/classificação , Filogenia , Proteínas não Estruturais Virais/genética , Proteínas Virais/genética
14.
Mem Inst Oswaldo Cruz ; 114: e190198, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31596309

RESUMO

BACKGROUND: In Brazil the implementation of the Sentinel Surveillance System of Influenza began in 2000. Central public health laboratories use reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for diagnosis of respiratory viruses, but this protocol identifies only specific targets, resulted in inconclusive diagnosis for many samples. Thus, high-throughput sequencing (HTS) would be complementary method in the identification of pathogens in inconclusive samples for RT-qPCR or other specific detection protocols. OBJECTIVES: This study aimed to detect unidentified viruses using HTS approach in negative samples of nasopharynx/tracheal secretions by the standard RT-qPCR collected in the Federal District, Brazil. METHODS: Nucleic acids were extracted from samples collected in winter period of 2016 and subjected to HTS. The results were confirmed by the multiplex PR21 RT-qPCR, which identifies 21 respiratory pathogens. FINDINGS: The main viruses identified by HTS were of families Herpesviridae, Coronaviridae, Parvoviridae and Picornaviridae, with the emphasis on rhinoviruses. The presence of respiratory viruses in the samples was confirmed by the PR21 multiplex RT-qPCR. Coronavirus, enterovirus, bocavirus and rhinovirus were found by multiplex RT-qPCR as well as by HTS analyses. MAIN CONCLUSIONS: Wide virus diversity was found by different methodologies and high frequency of rhinovirus occurrence was confirmed in population in winter, showing its relevance for public health.


Assuntos
Coronaviridae/isolamento & purificação , Herpesviridae/isolamento & purificação , Nasofaringe/virologia , Parvoviridae/isolamento & purificação , Picornaviridae/isolamento & purificação , Traqueia/virologia , Coronaviridae/classificação , Coronaviridae/genética , DNA Viral/genética , Herpesviridae/classificação , Herpesviridae/genética , Humanos , Parvoviridae/classificação , Parvoviridae/genética , Picornaviridae/classificação , Picornaviridae/genética , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
15.
Mol Cell Probes ; 48: 101448, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31521579

RESUMO

Amdoparvoviruses infect carnivore species, including mink, raccoon dog, fox, skunk, and red panda. Amdoparvovirus infection is a major cause of morbidity and mortality in farmed minks. Here, we developed a direct TaqMan qPCR assay for detection and quantification of carnivore amdoparvoviruses by using three primers and one probe based on the conserved VP2 gene. The detection limit for Aleutian mink disease virus (AMDV) and Raccoon dog and arctic fox amdoparvovirus (RFAV) were 4.06 × 101 copies/µl and 2.93 × 101 copies/µl, respectively. Both intra- and inter-assay variability were less than 2%. Among 74 carnivore samples, the positive rates for amdoparvoviruses were 62.2% (46/74) by direct TaqMan qPCR, while only 40.5% (30/74) by SYBR Green I qPCR. This result suggests that the direct TaqMan qPCR was more sensitive than the SYBR Green I qPCR. Additionally, the direct TaqMan qPCR is a rapid and sensitive method for liquid samples at microliter level as the assay employed the direct alkaline lysis method to obtain viral DNA and, therefore, eliminated the cumbersome steps in extracting DNA. Overall, the direct TaqMan qPCR assay possessed high specificity, sensitivity, and reproducibility, indicating that it can be used as a powerful tool for detection and quantification of various carnivore amdoparvoviruses in epidemiological and pathogenesis studies.


Assuntos
Vírus da Doença Aleutiana do Vison/genética , Parvoviridae/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Primers do DNA/genética , DNA Viral/genética , Cães , Raposas/virologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA/métodos
16.
Infect Genet Evol ; 75: 104013, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31446136

RESUMO

Bufavirus (BuV), a recent discovery virus of the family Parvoviridae, has been suggested to be a potential causative agent of diarrhea in humans. The prevalence of BuV has been demonstrated mostly in fecal specimens of diarrheic patients. Little is known about the presence of BuV in the environmental water. The aim of the present study was to conduct a surveillance for BuV in the environmental water in Thailand. A total of 125 water samples were collected during November 2016 and July 2018 from six different areas in Chiang Mai city, Thailand. Water samples were concentrated and extracted to obtain BuV genomic DNA. The BuV was screened by amplification of NS1 region using nested-PCR. The detected BuV was further characterized by amplification of VP2 region. The NS1 and VP2 genes were sequenced and analyzed phylogenetically. The BuV strain (CMW88/18) detected in this study belonged to BuV1 with the prevalence of 0.8%. The CMW88/18 strain was most closely related to human BuV1 strains reported previously worldwide suggesting contamination of BuV in the environmental water could be a potential source of infection in human.


Assuntos
Genoma Viral , Genômica , Parvoviridae/classificação , Parvoviridae/genética , Microbiologia da Água , Diarreia/epidemiologia , Diarreia/virologia , Genômica/métodos , Humanos , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Filogenia , Vigilância em Saúde Pública , Tailândia/epidemiologia
17.
Biologicals ; 61: 1-7, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31447377

RESUMO

Horses are often used as blood donors for commercial horse serum (HS) production and to manufacture biologicals. HS is an alternative for fetal bovine serum (FBS) used as a supplement for cell culture and vaccine production. Furthermore, HS is also frequently obtained in order to produce antisera toxins and pathogens. The advent of high-throughput sequencing (HTS) has promoted changes in virus detection, since previous knowledge of targets is not required. Thus, the present study aimed to describe the virome of five different batches of commercial HS from New Zealand (three batches) and Brazil and the United States (one batch each) using HTS. Each HS pool were processed and sequenced using an Illumina MiSeq platform. Sequences-related to viruses belonging to the Flaviviridae, Herpesviridae, and Parvoviridae families were detected. Particularly, equine hepacivirus (EqHV), equine pegivirus (EPgV), and Theiler's disease-associated virus (TDAV) were more frequent found in the batches analyzed. The presence of viral genomes in cell culture sera illustrates that these commercial sera can contain a mixture of different viruses and, therefore, can be regarded as potentially infectious for susceptible hosts. Moreover, the innocuity of commercial HS is important for the efficiency and security of diagnostics and the production of biological products.


Assuntos
Flaviviridae/genética , Genoma Viral , Herpesviridae/genética , Cavalos/virologia , Parvoviridae/genética , Soro/virologia , Animais , Meios de Cultura , Flaviviridae/classificação , Herpesviridae/classificação , Cavalos/sangue , Parvoviridae/classificação
18.
Viruses ; 11(6)2019 06 06.
Artigo em Inglês | MEDLINE | ID: mdl-31174309

RESUMO

Chapparvoviruses (ChPVs) comprise a divergent, recently identified group of parvoviruses (family Parvoviridae), associated with nephropathy in immunocompromised laboratory mice and with prevalence in deep sequencing results of livestock showing diarrhea. Here, we investigate the biological and evolutionary characteristics of ChPVs via comparative in silico analyses, incorporating sequences derived from endogenous parvoviral elements (EPVs) as well as exogenous parvoviruses. We show that ChPVs are an ancient lineage within the Parvoviridae, clustering separately from members of both currently established subfamilies. Consistent with this, they exhibit a number of characteristic features, including several putative auxiliary protein-encoding genes, and capsid proteins with no sequence-level homology to those of other parvoviruses. Homology modeling indicates the absence of a ß-A strand, normally part of the luminal side of the parvoviral capsid protein core. Our findings demonstrate that the ChPV lineage infects an exceptionally broad range of host species, including both vertebrates and invertebrates. Furthermore, we observe that ChPVs found in fish are more closely related to those from invertebrates than they are to those of amniote vertebrates. This suggests that transmission between distantly related host species may have occurred in the past and that the Parvoviridae family can no longer be divided based on host affiliation.


Assuntos
Especificidade de Hospedeiro , Invertebrados/virologia , Infecções por Parvoviridae/virologia , Parvovirus/classificação , Parvovirus/genética , Vertebrados/virologia , Animais , Proteínas do Capsídeo/genética , Evolução Molecular , Peixes/virologia , Genoma Viral , Parvoviridae/classificação , Parvoviridae/genética , Filogenia , Análise de Sequência , Homologia de Sequência , Proteínas Virais/genética , Sequenciamento Completo do Genoma
19.
Viruses ; 11(5)2019 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-31060205

RESUMO

Rodent protoparvoviruses (PVs), parvovirus H-1 (H-1PV) in particular, are naturally endowed with oncolytic properties. While being historically described as agents that selectively replicate in and kill cancer cells, recent yet growing evidence demonstrates that these viruses are able to reverse tumor-driven immune suppression through induction of immunogenic tumor cell death, and the establishment of antitumorigenic, proinflammatory milieu within the tumor microenvironment. This review summarizes the most important preclinical proofs of the interplay and the cooperation between PVs and the host immune system. The molecular mechanisms of PV-induced immunostimulation are also discussed. Furthermore, initial encouraging in-human observations from clinical trials and compassionate virus uses are presented, and speak in favor of further H-1PV clinical development as partner drug in combined immunotherapeutic protocols.


Assuntos
Neoplasias/virologia , Vírus Oncolíticos/fisiologia , Parvoviridae/fisiologia , Animais , Humanos , Sistema Imunitário , Neoplasias/imunologia , Neoplasias/terapia , Terapia Viral Oncolítica , Vírus Oncolíticos/genética , Parvoviridae/genética
20.
J Gen Virol ; 100(3): 367-368, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30672729

RESUMO

Members of the family Parvoviridae are small, resilient, non-enveloped viruses with linear, single-stranded DNA genomes of 4-6 kb. Viruses in two subfamilies, the Parvovirinae and Densovirinae, are distinguished primarily by their respective ability to infect vertebrates (including humans) versus invertebrates. Being genetically limited, most parvoviruses require actively dividing host cells and are host and/or tissue specific. Some cause diseases, which range from subclinical to lethal. A few require co-infection with helper viruses from other families. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the Parvoviridae, which is available at www.ictv.global/report/parvoviridae.


Assuntos
Infecções por Parvoviridae/virologia , Parvoviridae/classificação , Filogenia , Animais , Genoma Viral , Humanos , Parvoviridae/genética , Parvoviridae/isolamento & purificação , Parvoviridae/ultraestrutura , Virologia/organização & administração
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