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1.
Arch Virol ; 164(12): 2995-3006, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31576460

RESUMO

Reticuloendotheliosis virus (REV) is an important representative avian retrovirus. To improve our understanding of the host cellular responses to virus infection and the pathogenesis of REV infection, we applied isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with multidimensional liquid chromatography-tandem mass spectrometry to detect changes in protein levels in chicken embryo fibroblast cells (CEFs) that were infected with REV or mock infected. In total, 605 cellular proteins were differentially expressed, among which 196, 345, and 286 were differentially expressed in REV-infected CEFs at 1, 3, and 5 days postinfection, respectively. Gene Ontology analysis indicated that the biological processes of the differentially expressed proteins were primarily related to cellular processes, metabolic processes, biological regulation, response to stimulus, and immune system processes and that the molecular functions in which the differentially expressed proteins were mainly involved were binding, catalytic activity, and enzyme regulator activity. Pathway analysis showed that a total of 143, 167, and 179 pathways, including protein digestion and absorption, focal adhesion, ECM-receptor interaction, cytokine-cytokine receptor interaction, Toll-like receptors, and JAK-STAT signaling, were enriched in REV-infected CEFs at 1, 3, and 5 days postinfection, respectively. In conclusion, this study is the first to analyze the protein profile of REV-infected CEFs using an iTRAQ approach. The results of this study provide valuable information for better understanding the host response to REV infection.


Assuntos
Fibroblastos/metabolismo , Doenças das Aves Domésticas/genética , Proteoma/genética , Vírus da Reticuloendoteliose/fisiologia , Infecções por Retroviridae/veterinária , Animais , Embrião de Galinha , Galinhas , Fibroblastos/química , Fibroblastos/virologia , Doenças das Aves Domésticas/metabolismo , Doenças das Aves Domésticas/virologia , Proteoma/química , Proteoma/metabolismo , Proteômica , Vírus da Reticuloendoteliose/genética , Infecções por Retroviridae/genética , Infecções por Retroviridae/metabolismo , Infecções por Retroviridae/virologia , Espectrometria de Massas em Tandem
2.
BMC Infect Dis ; 19(1): 762, 2019 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-31477028

RESUMO

BACKGROUND: Avian influenza A (H5N6) virus poses a great threat to the human health since it is capable to cross the species barrier and infect humans. Although human infections are believed to largely originate from poultry contaminations, the transmissibility is unclear and only limited information was available on poultry environment contaminations, especially in Fujian Province. METHODS: A total of 4901 environmental samples were collected and tested for Avian Influenza Virus (AIV) from six cities in Fujian Province through the Fujian Influenza Surveillance System from 2013 to 2017. Two patient-related samples were taken from Fujian's first confirmed H5N6 human case and his backyard chicken feces in 2017. Chi-square test or Fisher's exact probability test was used to compare the AIV and the viral subtype positive rates among samples from different Surveillance cities, surveillance sites, sample types, and seasons. Phylogenetic tree analysis and molecular analysis were conducted to track the viral transmission route of the human infection and to map out the evolutions of H5N6 in Fujian. RESULTS: The overall positive rate of the H5 subtype AIVs was 4.24% (208/4903). There were distinctive differences (p < 0.05) in the positive rates in samples from different cities, sample sites, sample types and seasons. The viruses from the patient and his backyard chicken feces shared high homologies (99.9-100%) in all the eight gene segments. Phylogenetic trees also showed that these two H5N6 viruses were closely related to each other, and were classified into the same genetic clade 2.3.4.4 with another six H5N6 isolates from the environmental samples. The patient's H5N6 virus carried genes from H6N6, H5N8 and H5N6 viruses originated from different areas. The R294K or N294S substitution was not detected in the neuraminidase (NA). The S31 N substitution in the matrix2 (M2) gene was detected but only in one strain from the environmental samples. CONCLUSIONS: The H5 subtype of AIVs has started circulating in the poultry environments in Fujian Province. The patient's viral strain originated from the chicken feces in his backyard. Genetic reassortment in H5N6 viruses in Fujian Province was indicated. The H5N6 viruses currently circulating in Fujian Province were still commonly sensitive to Oseltamivir and Zanamivir, but the resistance against Amantadine has emerged.


Assuntos
Vírus da Influenza A/isolamento & purificação , Influenza Aviária/virologia , Influenza Humana/epidemiologia , Influenza Humana/virologia , Infecções por Orthomyxoviridae/virologia , Aves Domésticas/virologia , Animais , Embrião de Galinha , Galinhas/virologia , China/epidemiologia , Patos/virologia , Meio Ambiente , Microbiologia Ambiental , Genes Virais , Abrigo para Animais/normas , Humanos , Vírus da Influenza A/genética , Influenza Aviária/diagnóstico , Influenza Aviária/epidemiologia , Tipagem Molecular , Infecções por Orthomyxoviridae/diagnóstico , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/transmissão , Filogenia , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/virologia , Fatores de Risco
3.
Arch Virol ; 164(11): 2837-2841, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31494776

RESUMO

Since January 2019, abnormal molting has been observed frequently in approximately 40-day-old Pekin ducks in China. To investigate the possible involvement of a virus, we tested the prevalence of duck circovirus (DuCV), goose hemorrhagic polyomavirus (GHPyV), and goose parvovirus (GPV) in 11 molt cases in two provinces. GPV was detected in all cases, particularly in all samples collected from the feather area. The complete genome sequences of three GPV strains were determined and found to have 52 nucleotide changes relative to GPVs associated with short beak and dwarfism syndrome of Pekin ducks. These data will enhance our understanding of GPV diversity and outcomes of GPV infection in Pekin ducks.


Assuntos
Patos/virologia , Gansos/virologia , Muda/fisiologia , Parvovirinae/isolamento & purificação , Doenças das Aves Domésticas/virologia , Animais , China/epidemiologia , Circovirus/genética , Circovirus/isolamento & purificação , Genoma Viral/genética , Parvovirinae/genética , Polyomavirus/genética , Polyomavirus/isolamento & purificação , Doenças das Aves Domésticas/epidemiologia
4.
BMC Vet Res ; 15(1): 271, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31370846

RESUMO

BACKGROUND: Fowl adenoviruses (FAdVs) are associated with many diseases, resulting in huge economic losses to the poultry industry worldwide. Since 2015, outbreaks of FAdV infections with high mortality rates have been reported in China. A continued surveillance of FAdVs contributes to understand the epidemiology of the viruses. RESULTS: We isolated 155 FAdV strains from diseased chickens from poultry in China between 2015 and 2018. PCR analysis determined that 123 samples were FAdV species C, 27 were FAdV species E, and five contained two different FAdV strains. The phylogenetic analysis demonstrates that these sequences of hexon regions were clustered into three distinct serotypes: FAdV-4 (79.4%, 123/155), FAdV-8a (13.5%, 21/155) and FAdV-8b (3.9%, 6/155), of which FAdV-4 was the dominant serotype in China. CONCLUSIONS: The characterization of newly prevalent FAdV strains provides valuable information for the development of an effective control strategy for FAdV infections in chickens.


Assuntos
Infecções por Adenoviridae/veterinária , Aviadenovirus/classificação , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/virologia , Infecções por Adenoviridae/epidemiologia , Infecções por Adenoviridae/prevenção & controle , Infecções por Adenoviridae/virologia , Animais , Aviadenovirus/isolamento & purificação , Galinhas , China/epidemiologia , Filogenia , Doenças das Aves Domésticas/prevenção & controle , Sorogrupo
5.
BMC Vet Res ; 15(1): 274, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31370852

RESUMO

BACKGROUND: In Poland, the leader in goose production in Europe, goose parovirus infection, or Derzsy's disease (DD), must be reported to the veterinary administration due to the serious economic and epizootic threat to waterfowl production. Prophylactic treatment for DD includes attenuated live or inactivated vaccines. Moreover, the control of DD includes the monitoring of maternal derived antibody (MDA) levels in the offspring and antibody titers in the parent flock after vaccination. The aim of this study was to develop an ELISA for the detection of goose parvovirus (GPV) antibodies. RESULTS: Two recombinant protein fragments derived from VP3 (viral protein 3) GPV, namely VP3ep6 and VP3ep4-6 with a mass of 20.9 and 32.3 kDa, respectively, were produced using an Escherichia coli expression system. These proteins were purified by one-step nickel-affinity chromatography, which yielded protein preparations with a purity above 95%. These recombinant proteins were useful in the detection of serum anti-GPV antibodies, and this was confirmed by Western blotting. However, recombinant VP3ep4-6 protein showed a greater ability to correctly identify sera from infected geese. In the next stage of the project, a pool of 166 goose sera samples, previously examined by a virus neutralization test (VN), was tested. For further studies, one recombinant protein (VP3ep4-6) was selected for optimization of the test conditions. After optimization, the newly developed ELISA was compared to other serological tests, and demonstrated high sensitivity and specificity. CONCLUSION: In conclusion, the VP3ep4-6 ELISA method described here can be used for the detection of antibodies to GPV in serum.


Assuntos
Anticorpos Antivirais/sangue , Proteínas do Capsídeo/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Infecções por Parvoviridae/veterinária , Parvovirinae/imunologia , Doenças das Aves Domésticas/diagnóstico , Animais , Ensaio de Imunoadsorção Enzimática/normas , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/diagnóstico , Doenças das Aves Domésticas/sangue , Doenças das Aves Domésticas/virologia , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade
6.
BMC Vet Res ; 15(1): 253, 2019 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-31324180

RESUMO

BACKGROUND: Avian influenza virus (AIV), infectious bronchitis virus (IBV), and Newcastle disease virus (NDV) are important avian pathogens that can cause enormous economic loss on the poultry industry. Different respiratory etiological agents may induce similar clinical signs that make differential diagnosis difficult. Importantly, AIV brings about severe threat to human public health. Therefore, a novel method that can distinguish these viruses quickly and simultaneously is urgently needed. RESULTS: In this study, an oligonucleotide microarray system was developed. AIV, including H5, H7, and H9 subtypes; NDV; and IBV were simultaneously detected and differentiated on a microarray. Three probes specific for AIV, NDV, and IBV, as well as three other probes for differentiating H5, H7, and H9 of AIV, were first designed and jet-printed to predetermined locations of initiator-integrated poly(dimethylsiloxane) for the synchronous detection of the six pathogens. The marked multiplex reverse transcription polymerase chain reaction (PCR) products were hybridized with the specific probes, and the results of hybridization were read directly with the naked eyes. No cross-reaction was observed with 10 other subtypes of AIV and infectious bursal disease virus, indicating that the oligonucleotide microarray assay was highly specific. The sensitivity of the method was at least 100 times higher than that of the conventional PCR, and the detection limit of NDV, AIV, H5, H7, and H9 can reach 0.1 EID50 (50% egg infective dose), except that of IBV, which was 1 EID50 per reaction. In the validation of 93 field samples, AIV, IBV, and NDV were detected in 53 (56.99%) samples by oligonucleotide microarray and virus isolation and in 50 (53.76%) samples by conventional PCR. CONCLUSIONS: We have successfully developed an approach to differentiate AIV, NDV, IBV, H5, H7, and H9 subtypes of AIV using oligonucleotide microarray. The microarray is an accurate, high-throughput, and relatively simple method for the rapid detection of avian respiratory viral diseases. It can be used for the epidemiological surveillance and diagnosis of AIV, IBV, and NDV.


Assuntos
Vírus da Bronquite Infecciosa/genética , Vírus da Influenza A/genética , Vírus da Doença de Newcastle/genética , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Doenças das Aves Domésticas/virologia , Animais , Infecções por Coronavirus/virologia , Vírus da Doença Infecciosa da Bursa/genética , Influenza Aviária/virologia , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase Multiplex/veterinária , Doença de Newcastle/virologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Aves Domésticas , Sensibilidade e Especificidade
7.
BMC Vet Res ; 15(1): 256, 2019 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-31337397

RESUMO

BACKGROUND: Fowl adenovirus (FAdV) is an infectious agent that can cause jaundice, severe anaemia, dyspnoea and reproductive disorders in fowls. Since 2015, FAdV disease has been rapidly spreading among broiler chickens in China, where it has caused significant economic losses. In this study, a loop-mediated isothermal amplification (LAMP) real-time turbidity method with strong specificity to FAdV was established. RESULTS: The established assay was specific to FAdV-4, and the lower limit of detection was 75 copies/µL of extracted DNA. The positive detection rate for the suspected tissue samples was 33.3% (14/42), which was consistent with that of the real-time PCR. CONCLUSION: The proposed LAMP method can quickly and accurately detect prevalent FAdV via real-time turbidity assay, thereby providing a diagnostic platform for the prevention and control of the FAdV disease.


Assuntos
Infecções por Adenoviridae/veterinária , Adenoviridae/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/veterinária , Doenças das Aves Domésticas/virologia , Adenoviridae/genética , Infecções por Adenoviridae/diagnóstico , Infecções por Adenoviridae/virologia , Animais , Galinhas , DNA Viral , Técnicas de Amplificação de Ácido Nucleico/métodos , Doenças das Aves Domésticas/diagnóstico , Sensibilidade e Especificidade
8.
BMC Vet Res ; 15(1): 261, 2019 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-31345206

RESUMO

BACKGROUND: In Southeast Asian countries, including Myanmar, poultry farming is a major industry. In order to manage and maintain stable productivity, it is important to establish policies for biosecurity. Infectious respiratory diseases are a major threat to poultry farming. Avian influenza and Newcastle disease have been reported in Myanmar, but no scientific information is available for other respiratory pathogens, such as mycoplasmas and infectious bronchitis virus (IBV). Identifying the genotypes and serotypes of IBVs is especially important to inform vaccination programs. In this study, we detected Mycoplasma gallisepticum (MG), M. synoviae (MS), and IBV in several poultry farms in Myanmar. RESULTS: Samples were collected from 20 farms in three major poultry farming areas in Myanmar, and MG, MS, and IBV were detected on two, four, and eight farms, respectively, by polymerase chain reaction. Phylogenetic analysis revealed that the observed MG and MS isolates were not identical to vaccine strains. Three different genotypes of IBV were detected, but none was an unknown variant. CONCLUSIONS: Mycoplasmas and IBV were detected on poultry farms in Myanmar. Periodic surveillance is required to establish the distribution of each pathogen, and to institute better vaccine protocols.


Assuntos
Vírus da Bronquite Infecciosa/isolamento & purificação , Mycoplasma gallisepticum/isolamento & purificação , Mycoplasma synoviae/isolamento & purificação , Doenças das Aves Domésticas/epidemiologia , Animais , Galinhas , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Genótipo , Humanos , Vírus da Bronquite Infecciosa/genética , Mianmar/epidemiologia , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/veterinária , Mycoplasma gallisepticum/genética , Mycoplasma synoviae/genética , Filogenia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/virologia
9.
Res Vet Sci ; 125: 212-217, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31271953

RESUMO

Goose parvovirus (GPV) is the etiological agent of Derzsy's disease, with a natural reservoir consisting only of geese and Muscovy ducks. However, the pathological changes in the immune organs of ducklings experimentally infected with GPV remain unknown. In this study, 2-day-old Cherry Valley ducklings were intramuscularly injected with GPV. Immune organs (e.g., thymus, bursa of Fabricius, spleen, Harderian gland, cecal tonsil, bone marrow, and peripheral blood lymphocytes [PBLs]) were collected 1, 3, 5, 7, 10, and 14 days post-infection (dpi). Pathological lesions were assessed by histology and the viral load was concurrently assessed using quantitative real-time polymerase chain reaction. GPV antigen was detected via immunofluorescence staining and immunohistochemistry. No clinical symptoms or death were observed in the infected ducklings from 1 to 14 dpi; however, lesions with different degrees of hemorrhage and hyperemia were observed in the thymus, spleen and Harderian gland. Lymphocyte necrosis was identified in the thymus and spleen. In the immune organs, the highest viral loads were found in the spleen at 7 dpi, followed by the bone marrow, PBLs, and cecal tonsil at 3 dpi, and the bursa, Harderian gland, and thymus at 1 dpi. GPV antigen was primarily expressed in the cecal tonsil, spleen, and Harderian gland at 5 dpi, as well as in the PBLs and bone marrow at 3 dpi. Our findings indicate widespread GPV replication and dissemination in the immune organs of Cherry Valley ducklings.


Assuntos
Bolsa de Fabricius/patologia , Patos , Infecções por Parvoviridae/veterinária , Parvovirinae , Doenças das Aves Domésticas/virologia , Baço/patologia , Animais , Bolsa de Fabricius/virologia , Gansos , Infecções por Parvoviridae/patologia , Parvovirus/genética , Doenças das Aves Domésticas/patologia , Reação em Cadeia da Polimerase em Tempo Real , Baço/virologia , Carga Viral
10.
Onderstepoort J Vet Res ; 86(1): e1-e6, 2019 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-31291734

RESUMO

Between July and September 2017, samples collected from six unvaccinated chickens in Namibia were shown to be positive for infectious bursal disease virus (IBDV) by RT-PCR. Partial sequence and phylogenetic analysis of the VP1 and VP2 genes from six viruses revealed that they all belong to the very virulent pathotype (Genogroup 3) and are genetically very similar to IBDVs identified in neighbouring Zambia. This is the first molecular characterisation of IBDV in Namibia and has implications on the control and management of the disease in the country.


Assuntos
Infecções por Birnaviridae/veterinária , Galinhas , Vírus da Doença Infecciosa da Bursa/isolamento & purificação , Doenças das Aves Domésticas/epidemiologia , Animais , Infecções por Birnaviridae/epidemiologia , Vírus da Doença Infecciosa da Bursa/genética , Namíbia/epidemiologia , Filogenia , Doenças das Aves Domésticas/virologia , Análise de Sequência de DNA/veterinária
11.
Comp Immunol Microbiol Infect Dis ; 65: 110-115, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31300098

RESUMO

The coronavirus avian Infectious bronchitis virus (IBV) poses economic threats to poultry farms worldwide, affecting the performance of both meat-type and egg-laying birds. To define the evolution of recent IBVs in Iran, a genetic analysis based on hypervariable nucleotide sequences of S1 gene was carried out. Tracheal swab samples were collected from 170 Broiler flocks during 2017. Ten tracheal swabs from each flock pooled. From a total number of 170 flocks tested, 84.71% found to be positive. Phylogenetic tree analysis revealed the presence of D274 as a first time in Iran. IS/1494/06 was showed to be dominant IBV type circulating in broiler farms with a significantly higher prevalence than other four genotypes. Considering fluctuations in QX-type prevalence in recent years, continuous monitoring is necessary to reduce economic consequences in layer and broiler farms. The findings highlight the importance of using modified vaccination strategies that are adapted to the changing disease scenario.


Assuntos
Infecções por Coronavirus/veterinária , Genótipo , Vírus da Bronquite Infecciosa/genética , Filogenia , Doenças das Aves Domésticas/virologia , Animais , Galinhas , Infecções por Coronavirus/epidemiologia , Fazendas , Vírus da Bronquite Infecciosa/classificação , Vírus da Bronquite Infecciosa/isolamento & purificação , Irã (Geográfico)/epidemiologia , Doenças das Aves Domésticas/epidemiologia , Prevalência , Traqueia/virologia
12.
Comp Immunol Microbiol Infect Dis ; 65: 219-225, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31300117

RESUMO

The interaction between a low pathogenic avian influenza virus (A/CK/TUN/145/2012), a H9N2 Tunisian isolate, and a vaccine strain (H120) of avian infectious bronchitis, administered simultaneously or sequentially three days apart to chicks during 20 days, was evaluated using ELISA antibody levels, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses and histopathology examination. First, the in vivo replication interference of avian influenza virus (AIV) and infectious bronchitis virus (IBV) was evaluated using qRT-PCR to detect accurately either AIV or IBV genomes or viral copy numbers during dual infections. Second, we have determined the amount of specific antibodies in sera of chick's infected with AIV alone, IBV alone, mixed AIV + IBV, IBV then AIV or AIV IBV 3 days later using an ELISA test. Finally, histopathological analyses of internal organs from inoculated chicks were realized. Quantitative results of AIV and IBV co-infection showed that interferences between the two viruses yielded decreased viral growth. However, in the case of super-infection, the second virus, either AIV or IBV, induced a decrease in the growth of the first inoculated virus. According to our results, vaccine application was safe and do not interfere with AIV H9N2 infection, and does not enhance such infection. In conclusion, co-infection of chicks with AIV and IBV, simultaneously or sequentially, affected the clinical signs, the virus replication dynamics as well as the internal organ integrity. The results proposed that infection with heterologous virus may result in temporary competition for cell receptors or competent cells for replication, most likely interferon-mediated.


Assuntos
Coinfecção/veterinária , Vírus da Bronquite Infecciosa/crescimento & desenvolvimento , Vírus da Influenza A Subtipo H9N2/crescimento & desenvolvimento , Interferência Viral , Replicação Viral , Animais , Anticorpos Antivirais/sangue , Galinhas/imunologia , Galinhas/virologia , Coinfecção/virologia , Influenza Aviária/virologia , Doenças das Aves Domésticas/virologia , RNA Viral/análise , Vacinas Virais/imunologia
13.
Vet Microbiol ; 235: 136-142, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282371

RESUMO

Infectious Bursal Disease Virus (IBDV) of the ITA genotype (G6) was shown to have peculiar molecular characteristics and, despite a subclinical course, aggressiveness towards lymphoid tissues after experimental infection of specific-pathogen-free (SPF) chickens. The aim of the present study was to evaluate and compare with a Classical IBDV strain, ITA IBDV distribution and persistence in various tissues (bursa of Fabricious, spleen, thymus, bone marrow, caecal tonsils, Harderian gland, kidney, liver and proventriculus), its cloacal shedding and the involvement of gut TLR-3 in duodenum tissues. The 35-day-old SPF chickens were experimentally infected and sampled up to 28 days post infection (dpi) for IBDV detection and TLR-3 quantification by qRT-PCR. The ITA IBDV strain was detected in lymphoid and most non-lymphoid tissues up to the end of the trial, with higher loads compared to the Classical IBDV. Most of those differences were found during the first 2 weeks post-infection. Notably, bone marrow and caecal tonsils presented higher viral loads until 28 dpi, allowing to speculate that these organs may serve as non-bursal lymphoid tissues supporting virus replication. Differences in relative TLR-3 gene expression between ITA IBDV-infected birds and Classical-IBDV infected ones were observed at 4, 14 and 21 dpi, being initially higher in Classical group and later in ITA group. Our results provide new insights into IBDV pathogenesis showing that IBDV of ITA genotype leads to a high and persistent viral load in lymphoid tissues and to a delayed antiviral response.


Assuntos
Infecções por Birnaviridae/veterinária , Vírus da Doença Infecciosa da Bursa/genética , Tecido Linfoide/virologia , Doenças das Aves Domésticas/imunologia , Carga Viral , Animais , Infecções por Birnaviridae/imunologia , Medula Óssea/patologia , Medula Óssea/virologia , Galinhas , Ensaio de Imunoadsorção Enzimática , Genótipo , Vírus da Doença Infecciosa da Bursa/patogenicidade , Tonsila Palatina/virologia , Doenças das Aves Domésticas/virologia , Organismos Livres de Patógenos Específicos , Receptor 3 Toll-Like/genética , Replicação Viral
14.
Vet Microbiol ; 235: 151-163, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282373

RESUMO

This study demonstrates that the Muscovy duck reovirus (MDRV) p10.8 protein is one of many viral non-structural proteins that induces both cell cycle arrest and apoptosis. The p10.8 but not σC is a nuclear targeting protein that shuttles between the nucleus and the cytoplasm. Our results reveal that p10.8-induced apoptosis in cultured cells occurs by the nucleoporin Tpr/p53-dependent and Fas/caspase 8-mediated pathways. Furthermore, a compelling finding from this study is that the p10.8 and σC proteins of MDRV facilitate CDK2 and CDK4 degradation via the ubiquitin-proteasome pathway. We found that depletion of Cdc20 reversed the p10.8- and σC- mediated CDK4 degradation and p10.8-induced apoptosis, suggesting that Cdc20 plays a critical role in modulating p10.8-mediated cell cycle and apoptosis. Furthermore, we found that depletion of chaperonin-containing tailless complex polypeptide 1 (CCT) 2 and CCT5 reduced the level of Cdc20 and reversed the p10.8- and σC-mediated CDK4 degradation and p10.8-induced apoptosis, indicating that molecular chaperone CCT2 and CCT5 are required for stabilization of Ccd20 for mediating both cell cycle arrest and apoptosis. This study provides mechanistic insights into how p10.8 induces both cell cycle arrest and apoptosis.


Assuntos
Proteínas Cdc20/metabolismo , Chaperonina com TCP-1/metabolismo , Orthoreovirus/genética , Doenças das Aves Domésticas/virologia , Infecções por Reoviridae/veterinária , Proteínas não Estruturais Virais/metabolismo , Animais , Apoptose , Caspase 8/genética , Caspase 8/metabolismo , Proteínas Cdc20/genética , Pontos de Checagem do Ciclo Celular , Linhagem Celular , Chaperonina com TCP-1/genética , Patos/virologia , Fibroblastos/virologia , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , RNA Interferente Pequeno , Células Vero , Proteínas não Estruturais Virais/genética
15.
Vet Microbiol ; 235: 164-169, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282374

RESUMO

Marek's disease virus (MDV) is a highly contagious alphaherpesvirus that causes rapid onset of T cell lymphomas in chickens. MDV continues to break through vaccinal immunity due to the emergence of highly virulent field strains. Earlier studies revealed that deletion of the meq gene from MDV results in attenuated vaccines that protect against disease when chickens are infected with highly virulent strains. However, meq-deleted viruses still retain the ability to induce lymphoid organ atrophy, which raises safety concerns. In an earlier study, we found that deletion of lorf9 counteracts this lymphoid organ atrophy. Here, we describe the generation of a double deletion mutant virus lacking virus-encoded meq and lorf9. In vitro studies revealed that during replication, the mutant virus had kinetic characteristics similar to the parental virus; however, in vivo the replication capability was significantly reduced. Results of animal studies revealed no obvious MDV-specific symptoms and lesions. Importantly, the double deletion mutant virus lost the capacity to induce lymphoid organ atrophy, which has been the main obstacle during development of a good vaccine candidate.


Assuntos
Deleção de Genes , Herpesvirus Galináceo 1/genética , Herpesvirus Galináceo 1/patogenicidade , Tecido Linfoide/patologia , Doença de Marek/patologia , Proteínas Oncogênicas Virais/genética , Animais , Atrofia , Galinhas , Tecido Linfoide/virologia , Mutação , Doenças das Aves Domésticas/virologia , Replicação Viral
16.
Arch Virol ; 164(10): 2525-2530, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31286221

RESUMO

In this study, we investigated which structural proteins of pigeon paramyxovirus type 1 (PPMV-1) are responsible for its low pathogenicity in chickens. The results revealed that the pathogenicity of the virus is determined by multiple genes. The NP protein and F protein were found to have the strongest individual effect on virulence, and this effect further enhanced when the two proteins were expressed in combination. Our study highlights the influence of the NP and F proteins on the pathogenicity of PPMV-1 in chickens.


Assuntos
Doença de Newcastle/virologia , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/patogenicidade , Proteínas do Nucleocapsídeo/genética , Doenças das Aves Domésticas/virologia , Proteínas Virais de Fusão/genética , Fatores de Virulência/genética , Animais , Galinhas , Columbidae , Doença de Newcastle/patologia , Vírus da Doença de Newcastle/isolamento & purificação , Doenças das Aves Domésticas/patologia , Virulência
17.
BMC Vet Res ; 15(1): 232, 2019 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-31286975

RESUMO

BACKGROUND: Recently, serotype 4 fowl adenovirus (FAdV-4) has spread widely and caused huge economic loss to poultry industry. However, little is known about the molecular pathogenesis of FAdV-4. Fiber protein is thought to be vital for its infection and pathogenesis. RESULTS: Two novel monoclonal antibodies (mAbs) targeting the fiber-1 protein of FAdV-4 were generated, designated as mAb 3B5 and 6H9 respectively. Indirect immunofluorescence assay (IFA) showed that both mAbs only reacted with the FAdV-4 and FAdV-10, not with other serotypes including FAdV-1, FAdV-5, FAdV-6, FAdV-7, FAdV-8 and FAdV-9 tested. Although both mAbs did not recognize the linear epitopes, they could efficiently immunoprecipitate the fiber-1 protein in LMH cells either infected with FAdV-4 or transfected with pcDNA3.1-Fiber-1. Moreover, mAb 3B5 as a capture antibody and HRP-conjugated mAb 6H9 as a detection antibody, a novel sandwich ELISA for efficient detection of FAdV-4 was generated. The limit of detection of the ELISA could reach to 1000 TCID50/ml of FAdV-4 and the ELISA could be efficiently applied to detect FAdV-4 in the clinical samples. CONCLUSION: The two mAbs specific targeting fiber-1 generated here would pave the way for further studying on the role of fiber-1 in the infection and pathogenesis of FAdV-4, and the established mAb based sandwich ELISA would provide an efficient diagnostics tool for detection of FAdV-4/10.


Assuntos
Infecções por Adenoviridae/diagnóstico , Anticorpos Monoclonais/metabolismo , Aviadenovirus/fisiologia , Proteínas do Capsídeo/imunologia , Doenças das Aves Domésticas/diagnóstico , Infecções por Adenoviridae/virologia , Animais , Anticorpos Antivirais/metabolismo , Aviadenovirus/genética , Proteínas do Capsídeo/genética , Linhagem Celular , Galinhas , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Limite de Detecção , Camundongos Endogâmicos BALB C , Doenças das Aves Domésticas/virologia
18.
Vet Microbiol ; 233: 1-4, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31176393

RESUMO

Domestic ducks are considered as the interface between wild aquatic birds and terrestrial poultry and play an important role in the transmission and evolution of avian influenza viruses (AIVs). However, the infectivity of H9N2 AIVs in different domestic duck species has not been systematically evaluated. Here we investigated the infectivity of various genotypes of chicken H9N2 AIVs in Pekin duck (Anas Platyrhynchos), Mallard duck (Anas Platyrhynchos) and Muscovy duck (Cairina Moschata) through intranasal inoculation. We found that Pekin ducks and Mallard ducks were generally resistant to chicken H9N2 virus infection, while Muscovy ducks were relatively susceptible to H9N2 AIVs. All the tested viruses were isolated from oropharynx, trachea and lung tissues of Muscovy ducks. Additionally, genotype 57 (G57) H9N2 AIVs, which was predominant in chickens since 2010, showed increased virus replication in this duck species, indicating an improved interspecies transmission ability of recent H9N2 viruses from chickens to ducks. Our results demonstrated the role of Muscovy ducks in the ecology of H9N2 AIVs. More attentions should be paid to this host during viral surveillances. Additionally, inactivated H9N2 vaccine may be unnecessarily used in Pekin and Mallard ducks.


Assuntos
Patos/virologia , Influenza Aviária/transmissão , Doenças das Aves Domésticas/virologia , Replicação Viral , Animais , Galinhas/virologia , Suscetibilidade a Doenças , Vírus da Influenza A Subtipo H9N2/genética , Vírus da Influenza A Subtipo H9N2/fisiologia , Pulmão/virologia , Orofaringe/virologia , Traqueia/virologia
19.
Vet Microbiol ; 233: 85-92, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31176417

RESUMO

Muscovy duck reovirus (MDRV) causes serious immunodeficiency in the intestinal mucosa, although the underlying histopathological mechanisms remain unclear. Thus, we investigated the impact of MDRV infection on intestinal morphology using hematoxylin and eosin staining. Immune-related cells were also quantified by staining with hematoxylin and eosin, toluidine blue, and periodic acid-Schiff stain, or by immunohistochemistry and cytochemistry for lectin. Similarly, CD4+ and CD8+ cells were quantified by flow cytometry, and the expression of several immune-related molecules was quantified by radioimmunoassay. We found that MDRV clearly damaged the intestinal mucosa, based on tissue morphology, villus length, villus width, intestinal thickness, villus height/crypt depth ratio, and villus surface area. MDRV also altered the density or distribution of lymphocytes, mastocytes, and goblet cells in the small intestinal mucosa, as well as microfold cells in Peyer's patches. In addition, MDRV markedly depleted CD4+ cells from the intestinal mucosa and lowered the CD4+:CD8+ ratio in peripheral blood. Moreover, MDRV diminished the levels of secretory IgA and mucosal addressin cell adhesion molecule-1 (p < 0.01), but elevated those of histamine and nitric oxide (p < 0.01 or p < 0.05). Finally, MDRV significantly suppressed IL-1ß, IL-4, IL-5, and IL-8 levels (p < 0.01 or p < 0.05) mid-infection. Collectively, our data suggest that MDRV severely damages the structure and function of the intestinal mucosa by modulating immune cells and immune-related factors, thus leading to local immunodeficiency. Our findings lay the foundation for further research on the pathogenesis of MDRV.


Assuntos
Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Intestino Delgado/virologia , Orthoreovirus Aviário/imunologia , Infecções por Reoviridae/imunologia , Fatores Etários , Animais , Contagem de Linfócito CD4 , Citocinas/imunologia , Patos/virologia , Duodeno , Fibroblastos/virologia , Histamina/análise , Imunoglobulina A Secretora/análise , Intestino Delgado/imunologia , Óxido Nítrico/análise , Orthoreovirus Aviário/patogenicidade , Doenças das Aves Domésticas/virologia , Infecções por Reoviridae/patologia , Carga Viral
20.
Infect Dis Poverty ; 8(1): 56, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31230595

RESUMO

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by severe fever with thrombocytopenia syndrome virus (SFTSV). SFTSV has been found in humans, ticks and animals, and SFTS has high mortality and increasing prevalence in East Asia. In the study, the samples (heart, liver, lung, kidney, spleen, brain tissue and serum) were collected from 374 domestic animals and 241 wild animals in Pingqiao District and Xinxian County of Xinyang in Henan Province, China. 275 (44.72%, 275/615) animals were positive for anti-SFTSV antibodies, the anti-SFTSV antibodies positive ratios of domestic and wild animals were 43.58% (163/374) and 46.47% (112/241), respectively. There was no significant difference in domestic and wild animals, but significant differences were detected among different species of animals (χ2 = 112.59, P < 0.0001). Among 615 animals, 105 (17.07%, 105/615) animals were positive for SFTSV RNA, and only one SFTSV strain was isolated from heart tissue of a yellow weasel. The phylogenetic analysis shows that the sequence from animals belonged to the same group with viral sequences obtained from humans. The animals maybe play a reservoir host in maintaining the life cycle of SFTSV in nature.


Assuntos
Doenças dos Animais/epidemiologia , Infecções por Bunyaviridae/veterinária , Phlebovirus/isolamento & purificação , Doenças dos Animais/virologia , Animais , Aves , Infecções por Bunyaviridae/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Galinhas , China/epidemiologia , Doenças do Cão/epidemiologia , Doenças do Cão/virologia , Cães , Patos , Mamíferos , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/virologia , Prevalência , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/virologia , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/virologia
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