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1.
Viruses ; 14(5)2022 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-35632659

RESUMO

Commercial inactivated vaccines against H9N2 avian influenza (AI) have been developed in China since 1990s and show excellent immunogenicity with strong HI antibodies. However, currently approved vaccines cannot meet the clinical demand for a live-vectored vaccine. Newcastle disease virus (NDV) vectored vaccines have shown effective protection in chickens against H9N2 virus. However, preexisting NDV antibodies may affect protective efficacy of the vaccine in the field. Here, we explored avian paramyxovirus serotype 2 (APMV-2) as a vector for developing an H9N2 vaccine via intranasal delivery. APMV-2 belongs to the same genus as NDV, distantly related to NDV in the phylogenetic tree, based on the sequences of Fusion (F) and hemagglutinin-neuraminidase (HN) gene, and has low cross-reactivity with anti-NDV antisera. We incorporated hemagglutinin (HA) of H9N2 into the junction of P and M gene in the APMV-2 genome by being flanked with the gene start, gene end, and UTR of each gene of APMV-2-T4 to generate seven recombinant APMV-2 viruses rAPMV-2/HAs, rAPMV-2-NPUTR-HA, rAPMV-2-PUTR-HA, rAPMV-2-FUTR-HA, rAPMV-2-HNUTR-HA, rAPMV-2-LUTR-HA, and rAPMV-2-MUTR-HA, expressing HA. The rAPMV-2/HAs displayed similar pathogenicity compared with the parental APMV-2-T4 virus and expressed HA protein in infected CEF cells. The NP-UTR facilitated the expression and secretion of HA protein in cells infected with rAPMV-2-NPUTR-HA. Animal studies demonstrated that immunization with rAPMV-2-NPUTR-HA elicited effective H9N2-specific antibody (6.14 ± 1.2 log2) responses and conferred complete immune protection to prevent viral shedding in the oropharyngeal and cloacal swabs from chickens challenged with H9N2 virus. This study suggests that our recombinant APMV-2 virus is safe and immunogenic and can be a useful tool in the combat of H9N2 outbreaks in chicken.


Assuntos
Avulavirus , Vírus da Influenza A Subtipo H9N2 , Influenza Aviária , Animais , Galinhas , Hemaglutininas , Imunização , Vírus da Influenza A Subtipo H9N2/genética , Vírus da Doença de Newcastle/genética , Filogenia , Sorogrupo , Vacinas Atenuadas , Vacinas Sintéticas/genética
2.
Appl Environ Microbiol ; 88(11): e0046622, 2022 06 14.
Artigo em Inglês | MEDLINE | ID: mdl-35612300

RESUMO

Avian paramyxoviruses (APMVs) (subfamily Avulavirinae) have been isolated from over 200 species of wild and domestic birds around the world. The International Committee on Taxonomy of Viruses (ICTV) currently defines 22 different APMV species, with Avian orthoavulavirus 1 (whose viruses are designated APMV-1) being the most frequently studied due to its economic burden to the poultry industry. Less is known about other APMV species, including limited knowledge on the genetic diversity in wild birds, and there is a paucity of public whole-genome sequences for APMV-2 to -22. The goal of this study was to use MinION sequencing to genetically characterize APMVs isolated from wild bird swab samples collected during 2016 to 2018 in the United States. Multiplexed MinION libraries were prepared using a random strand-switching approach using 37 egg-cultured, influenza-negative, hemagglutination-positive samples. Forty-one APMVs were detected, with 37 APMVs having complete polymerase coding sequences allowing for species identification using ICTV's current Paramyxoviridae phylogenetic methodology. APMV-1, -4, -6, and -8 viruses were classified, one putative novel species (Avian orthoavulavirus 23) was identified from viruses isolated in this study, two putative new APMV species (Avian metaavulavirus 24 and 27) were identified from viruses isolated in this study and from retrospective GenBank sequences, and two putative new APMV species (Avian metaavulavirus 25 and 26) were identified solely from retrospective GenBank sequences. Furthermore, coinfections of APMVs were identified in four samples. The potential limitations of the branch length being the only species identification criterion and the potential benefit of a group pairwise distance analysis are discussed. IMPORTANCE Most species of APMVs are understudied and/or underreported, and many species were incidentally identified from asymptomatic wild birds; however, the disease significance of APMVs in wild birds is not fully determined. The rapid rise in high-throughput sequencing coupled with avian influenza surveillance programs have identified 12 different APMV species in the last decade and have challenged the resolution of classical serological methods to identify new viral species. Currently, ICTV's only criterion for Paramyxoviridae species classification is the requirement of a branch length of >0.03 using a phylogenetic tree constructed from polymerase (L) amino acid sequences. The results from this study identify one new APMV species, propose four additional new APMV species, and highlight that the criterion may have insufficient resolution for APMV species demarcation and that refinement or expansion of this criterion may need to be established for Paramyxoviridae species identification.


Assuntos
Animais Selvagens , Infecções por Avulavirus , Avulavirus , Doenças das Aves , Animais , Animais Selvagens/virologia , Avulavirus/genética , Avulavirus/isolamento & purificação , Infecções por Avulavirus/epidemiologia , Infecções por Avulavirus/veterinária , Infecções por Avulavirus/virologia , Doenças das Aves/epidemiologia , Doenças das Aves/virologia , Aves , Filogenia , Estudos Retrospectivos , Vigilância de Evento Sentinela/veterinária , Estados Unidos/epidemiologia
3.
Microbiol Spectr ; 10(2): e0206121, 2022 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-35357204

RESUMO

Avian paramyxovirus 1 (APMV-1), also known as Newcastle disease virus (NDV), causes severe and economically important disease in poultry around the globe. Although a limited amount of APMV-1 strains in urban areas have been characterized, the role of the urban wild bird population as an APMV-1 reservoir is unclear. Because urban birds may have an important role for long-term circulation of the virus, fecal and swab samples were collected by community scientists from wild birds in New York City (NYC), New York, United States. These samples were screened for APMV-1 and genotypically characterized by sequencing of the complete genome. A total of 885 samples were collected from NYC parks and from a local wildlife rehabilitation clinic from October 2020 through June 2021, and 255 samples obtained from 197 birds have been processed to date. Eight birds (4.1%) screened positive for the APMV-1 nucleoprotein gene by conventional reverse transcription PCR (RT-PCR), and two live viruses were isolated via egg culture. A multibasic F protein cleavage sequence, 112R R K K R F117, an indicator of highly pathogenic velogenic APMV-1 strains, was present in the two samples fully sequenced by next generation sequencing. Phylogenetic analysis of the F gene coding sequence classified both isolates into genotype VI, a diverse and predominant genotype responsible for APMV-1 outbreaks in pigeon and dove species worldwide. IMPORTANCE Here we describe the first large-scale effort to screen for APMV-1 in New York City's wild bird population as part of the New York City Virus Hunters program, a community science initiative. We characterized two isolates of APMV-1, with phylogenetic analyses suggesting diversity in established and circulating strains of pigeon paramyxoviruses. Our isolates are also domestic reference strains for future APMV-1 vaccine developments. Future surveillance in this region may contribute to our understanding of APMV-1's evolution and genetic diversity, as well as inform poultry husbandry and vaccination practices in New York State.


Assuntos
Avulavirus , Doença de Newcastle , Animais , Animais Selvagens , Avulavirus/genética , Columbidae , Cidade de Nova Iorque/epidemiologia , Doença de Newcastle/epidemiologia , Vírus da Doença de Newcastle/genética , Filogenia , Aves Domésticas , Estados Unidos
4.
J Vet Med Sci ; 84(3): 378-389, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-35082196

RESUMO

Avian paramyxoviruses (APMVs) belonging to the subfamily Avulavirinae within the family Paramyxoviridae. APMVs consist of twenty-two known species and are constantly isolated from a wide variety of avian species around the world. In this study, the APMV isolates obtained from wild birds and domestic poultry during 2009-2020 in Taiwan were genetically characterized by phylogenetic analysis of their complete fusion protein gene or full-length genome. As a result, 57 APMV isolates belonging to seven different species were obtained during this period and subsequently identified as APMV-1 (n=17), APMV-2 (n=1), APMV-4 (n=25), APMV-6 (n=8), APMV-12 (n=2), APMV-21 (n=2) and APMV-22 (n=2). Sanger sequencing was performed to provide 22 full-length genome sequences and 35 complete fusion protein gene sequences for the APMV isolates. Phylogenetic analysis showed that the recovered viruses were closely related to Eurasian strains, except five class I APMV-1 and four APMV-4 isolates were related to North America strains. Our findings provided more evidence for the intercontinental transmission of APMVs between Eurasia and North America by wild birds. In addition, according to the criteria of the classification system based on complete fusion protein gene sequences, three novel genotypes within APMV-2, APMV-12, and APMV-22 were identified. Together, this investigation provided a broader perspective on the genetic diversity, evolution, and distribution of APMVs in multiple avian host species sampled in Taiwan.


Assuntos
Avulavirus , Animais , Avulavirus/genética , Aves , Variação Genética , Filogenia , Aves Domésticas , Taiwan/epidemiologia
5.
Vet Res Commun ; 46(1): 159-168, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34580815

RESUMO

Avian orthoavulavirus 13 (AOAV-13), formerly known as Avian paramyxovirus 13 (APMV-13), is found scatteredly in wild birds around the world. Although four complete genome sequences of AOAV-13 had been identified since the first discovery in Japan in 2003, the information available on the genetic variation and biological characteristics of AOAV-13 is still limited. In the present study, we isolated six AOAV-13 strains from fecal samples of wild migratory waterfowls during annual (2014-2018) viral surveillance of wild bird populations from wetland and domestic poultry of live bird markets (LBMs) in China. The phylogenetic analyses based on the HN and F genes showed that they had very close relationship and the molecular clock estimations showed a low evolutionary rate of AOAV-13. However, Bean goose/Hubei/V97-1/2015 is 1953 nt in size (ORF, 1, 776 nt), which is a unique size and longer than other reported AOAV-13 strains. Additionally, four repeats of conserved sequences "AAAAAT" was presented in the 5'-end trailer region of Swan goose/Hubei/VI49-1/2016, which is unprecedented in the AOAV-13. These findings highlight the importance of continuous monitoring the specific species of APMVs.


Assuntos
Infecções por Avulavirus , Avulavirus , Doenças das Aves Domésticas , Animais , Infecções por Avulavirus/veterinária , Galinhas , Filogenia
6.
Methods Mol Biol ; 2411: 63-73, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34816398

RESUMO

Avian paramyxoviruses (APMVs) have gained a great attention to be developed as vaccine vectors against human and veterinary pathogens. Avirulent APMVs are highly safe to be used as vaccine vectors for avian and non-avian species. APMV vectored vaccines induce robust cellular and humoral immune responses in a broad range of hosts. APMV vectors can be a good platform by facilitating rapid generation of vaccines against emerging pathogens. In this chapter, we discuss application of reverse genetics of APMVs for vaccine development, design of APMV vectored vaccines, cloning of protective antigen(s) into a vector, recovery of vectored vaccines and characterization of generated vaccine viruses.


Assuntos
Avulavirus , Avulavirus/genética , Vetores Genéticos/genética , Humanos
7.
Avian Dis ; 65(1): 63-66, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-34339124

RESUMO

Minnesota is the leading state in number of turkeys produced in the United States. Turkey flocks in the field are usually vaccinated several times with live avian orthoavulavirus 1 (AOAV-1) vaccines starting as early as 2 wk of age (WOA). During the years 2018-2019, many turkey flocks were diagnosed with low-virulence AOAV-1 infection around 9 WOA that led to respiratory disease, although they were previously vaccinated. This study was designed to investigate the immunity against AOAV-1 in Minnesota turkey flocks in the field and experimentally after vaccination. We reviewed antibody titers against AOAV-1 from turkey flocks tested by ELISA at Minnesota Poultry Testing Laboratory (n = 1292). Up to 9 WOA, more than 85% of the field flocks tested had unprotective antibody titers against AOAV-1. However, commercial poults at 3 WOA experimentally vaccinated by eye-drop method had an ELISA geometric mean titer of 6011 at 7 WOA. Oropharyngeal virus shedding after vaccination was 10%, 70%, 80%, and 40% at 1, 3, 5, and 7 days postvaccination, respectively. This study demonstrates that experimentally vaccinated turkeys respond very well to AOAV-1 vaccine when properly administered. However, there is clear vaccination failure in the field, where vaccine is commonly administered in drinking water, a method that is more susceptible to failure because of many variables in this procedure. We recommend choosing the most effective method of vaccine administration. Given the high incidence of inadequate immunity induced in commercial turkeys on mass application of live AOAV-1 vaccines in water, alternative application methods and subsequent monitoring of the serologic antibody response must be undertaken to ensure a proper immune response.


Artículo regular­Fracaso de la vacunación contra el Orthoavulavirus aviar 1 en pavos de Minnesota. Minnesota es el estado líder en número de pavos producidos en los Estados Unidos. Las parvadas de pavos en el campo generalmente se vacunan varias veces con vacunas vivas con Orthoavulavirus Aviar 1 (AOAV-1) comenzando desde las 2 semanas de edad (WOA). Durante los años 2018­2019, muchas parvadas de pavos fueron diagnosticadas con infección por Orthoavulavirus Aviar 1 de baja virulencia alrededor de las nueve semanas de edad que condujeron a una enfermedad respiratoria, aunque las aves fueron vacunadas previamente. Este estudio fue diseñado para investigar la inmunidad contra Orthoavulavirus Aviar 1 en parvadas de pavos de Minnesota en el campo y experimentalmente después de la vacunación. Se revisaron los títulos de anticuerpos contra Orthoavulavirus Aviar 1 de parvadas de pavos analizados por ELISA en el Laboratorio de Diagnóstico Avícola de Minnesota (n = 1292). Hasta las nueve semanas de edad, más del 85% de las parvadas de campo analizadas tenían títulos de anticuerpos no protectores contra Orthoavulavirus Aviar 1. Sin embargo, los pavipollos comerciales a las tres semanas de edad vacunados experimentalmente por el método de gota ocular tenían un título medio geométrico de ELISA de 6011 a las siete semanas de edad. La diseminación del virus orofaríngeo después de la vacunación fue del 10%, 70%, 80% y 40% a los 1, 3, 5 y 7 días después de la vacunación, respectivamente. Este estudio demuestra que los pavos vacunados experimentalmente respondieron muy bien a la vacuna con Orthoavulavirus Aviar 1 cuando se administra correctamente. Sin embargo, existe un claro fracaso de la vacunación en el campo, donde la vacuna se administra comúnmente en el agua potable, un método que es más susceptible al fracaso debido a muchas variables en este procedimiento. Se recomienda elegir el método de administración de vacunas más eficaz. Considerando la alta incidencia de inmunidad inadecuada inducida en pavos comerciales con la aplicación masiva de vacunas vivas con Orthoavulavirus Aviar 1 en agua, se deben llevar a cabo métodos de aplicación alternativos y monitoreo posterior de la respuesta de anticuerpos serológicos para asegurar una respuesta inmune adecuada.


Assuntos
Infecções por Avulavirus/veterinária , Avulavirus/efeitos dos fármacos , Doenças das Aves Domésticas/prevenção & controle , Falha de Tratamento , Perus , Vacinação/veterinária , Vacinas Virais/administração & dosagem , Animais , Infecções por Avulavirus/prevenção & controle , Infecções por Avulavirus/virologia , Minnesota , Doenças das Aves Domésticas/virologia
8.
Viruses ; 13(4)2021 03 24.
Artigo em Inglês | MEDLINE | ID: mdl-33805157

RESUMO

We describe for the first time the genetic and antigenic characterization of 18 avian avulavirus type-6 viruses (AAvV-6) that were isolated from wild waterfowl in the Americas over the span of 12 years. Only one of the AAvV-6 viruses isolated failed to hemagglutinate chicken red blood cells. We were able to obtain full genome sequences of 16 and 2 fusion gene sequences from the remaining 2 isolates. This is more than double the number of full genome sequences available at the NCBI database. These AAvV-6 viruses phylogenetically grouped into the 2 existing AAvV-6 genotype subgroups indicating the existence of an intercontinental epidemiological link with other AAvV-6 viruses isolated from migratory waterfowl from different Eurasian countries. Antigenic maps made using HI assay data for these isolates showed that the two genetic groups were also antigenically distinct. An isolate representing each genotype was inoculated in specific pathogen free (SPF) chickens, however, no clinical symptoms were observed. A duplex fusion gene based real-time assay for the detection and genotyping of AAvV-6 to genotype 1 and 2 was developed. Using the developed assay, the viral shedding pattern in the infected chickens was examined. The chickens infected with both genotypes were able to shed the virus orally for about a week, however, no significant cloacal shedding was detected in chickens of both groups. Chickens in both groups developed detectable levels of anti-hemagglutinin antibodies 7 days after infection.


Assuntos
Animais Selvagens/virologia , Antígenos Virais/imunologia , Infecções por Avulavirus/veterinária , Avulavirus/genética , Doenças das Aves/epidemiologia , Doenças das Aves/virologia , Genótipo , Migração Animal , Animais , Avulavirus/classificação , Avulavirus/imunologia , Avulavirus/isolamento & purificação , Doenças das Aves/transmissão , Canadá/epidemiologia , Galinhas/virologia , Cloaca/virologia , Genoma Viral , Testes de Hemaglutinação , Filogenia , Doenças das Aves Domésticas/virologia , Organismos Livres de Patógenos Específicos , Eliminação de Partículas Virais
9.
Viruses ; 13(3)2021 03 08.
Artigo em Inglês | MEDLINE | ID: mdl-33800329

RESUMO

Significant mortalities of racing pigeons occurred in Australia in late 2011 associated with a pigeon paramyxovirus serotype 1 (PPMV-1) infection. The causative agent, designated APMV-1/pigeon/Australia/3/2011 (P/Aus/3/11), was isolated from diagnostic specimens in specific pathogen free (SPF) embryonated eggs and was identified by a Newcastle Disease virus (NDV)-specific RT-PCR and haemagglutination inhibition (HI) test using reference polyclonal antiserum specific for NDV. The P/Aus/3/11 strain was further classified as PPMV-1 using the HI test and monoclonal antibody 617/161 by HI and phylogenetic analysis of the fusion gene sequence. The isolate P/Aus/3/11 had a slow haemagglutin-elution rate and was inactivated within 45 min at 56 °C. Cross HI tests generated an R value of 0.25, indicating a significant antigenic difference between P/Aus/3/11 and NDV V4 isolates. The mean death time (MDT) of SPF eggs infected with the P/Aus/3/11 isolate was 89.2 hr, characteristic of a mesogenic pathotype, consistent with other PPMV-1 strains. The plaque size of the P/Aus/3/11 isolate on chicken embryo fibroblast (CEF) cells was smaller than those of mesogenic and velogenic NDV reference strains, indicating a lower virulence phenotype in vitro and challenge of six-week-old SPF chickens did not induce clinical signs. However, sequence analysis of the fusion protein cleavage site demonstrated an 112RRQKRF117 motif, which is typical of a velogenic NDV pathotype. Phylogenetic analysis indicated that the P/Aus/3/11 isolate belongs to a distinct subgenotype within class II genotype VI of avian paramyxovirus type 1. This is the first time this genotype has been detected in Australia causing disease in domestic pigeons and is the first time since 2002 that an NDV with potential for virulence has been detected in Australia.


Assuntos
Avulavirus/genética , Avulavirus/isolamento & purificação , Columbidae/virologia , Genoma Viral , Genótipo , Filogenia , Animais , Avulavirus/classificação , Avulavirus/patogenicidade , Galinhas/virologia , Testes de Inibição da Hemaglutinação , Organismos Livres de Patógenos Específicos , Vitória , Proteínas Virais de Fusão/genética , Proteínas Virais de Fusão/imunologia , Virulência , Zigoto/virologia
10.
Viruses ; 13(2)2021 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-33669530

RESUMO

A reverse genetic system for avian paramyxovirus type-3 (APMV-3) strain Wisconsin was created and the infectious virus was recovered from a plasmid-based viral antigenomic cDNA. Green fluorescent protein (GFP) gene was cloned into the recombinant APMV-3 genome as a foreign gene. Stable expression of GFP by the recovered virus was confirmed for at least 10 consecutive passages. APMV-3 strain Wisconsin was evaluated against APMV-3 strain Netherlands and APMV-1 strain LaSota as a vaccine vector. The three viral vectors expressing GFP as a foreign protein were compared for level of GFP expression level, growth rate in chicken embryo fibroblast (DF-1) cells, and tissue distribution and immunogenicity in specific pathogen-free (SPF) day-old chickens. APMV-3 strain Netherlands showed highest growth rate and GFP expression level among the three APMV vectors in vitro. APMV-3 strain Wisconsin and APMV-1 strain LaSota vectors were mainly confined to the trachea after vaccination of day-old SPF chickens without any observable pathogenicity, whereas APMV-3 strain Netherlands showed wide tissue distribution in different body organs (brain, lungs, trachea, and spleen) with mild observable pathogenicity. In terms of immunogenicity, both APMV-3 strain-vaccinated groups showed HI titers two to three fold higher than that induced by APMV-1 strain LaSota vaccinated group. This study offers a novel paramyxovirus vector (APMV-3 strain Wisconsin) which can be used safely for vaccination of young chickens as an alternative for APMV-1 strain LaSota vector.


Assuntos
Infecções por Avulavirus/veterinária , Avulavirus/genética , Vetores Genéticos/genética , Doenças das Aves Domésticas/virologia , Vacinas Virais/genética , Animais , Avulavirus/metabolismo , Infecções por Avulavirus/prevenção & controle , Infecções por Avulavirus/virologia , Galinhas , Vetores Genéticos/metabolismo , Doenças das Aves Domésticas/prevenção & controle , Genética Reversa , Organismos Livres de Patógenos Específicos , Vacinas Virais/administração & dosagem , Vacinas Virais/imunologia , Wisconsin
11.
Trop Anim Health Prod ; 53(1): 90, 2021 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-33415381

RESUMO

Circulation of the dominant sub-genotype VII.2 of Avian Orthoavulavirus-1 (AOAV-1) is affecting multiple poultry and non-poultry avian species and causing significant economic losses to the poultry industry worldwide. In countries where ND is endemic, continuous monitoring and characterization of field strains are necessary. In this study, genetic characteristics of eleven AOAV-1 strains were analyzed isolated from wild birds including parakeets (n = 3), lovebird parrot (n = 1), pheasant (n = 1), peacock (n = 1), and backyard chickens (n = 5) during 2015-2016. Genetic characterization (genome size [15,192 nucleotides], the presence of typical cleavage site [112-RRQKRF-117]) and biological assessment (HA log 27 to 29 and intracerebral pathogenicity index [ICPI] value ranging from 1.50 to 1.86) showed virulent AOAV-1. Phylogenetic analysis showed that the studied isolates belonged to sub-genotype VII.2 and genetically very closely related (> 98.9%) to viruses repeatedly isolated (2011-2018) from commercial poultry. These findings provide evidence for the existence of epidemiological links between poultry and wild bird species in the region where the disease is prevalent. The deduced amino acid analysis revealed several substitutions in critical domains of fusion and hemagglutinin-neuraminidase genes. The pathogenesis and transmission potential of wild bird-origin AOAV-1 strain (AW-Pht/2015) was evaluated in 21-day-old chickens that showed the strain was highly virulent causing clinical signs and killed all chickens. High viral loads were detected in different organs of the infected chickens correlating with the severity of lesions developed. The continuous monitoring of AOAV-1 isolates in different species of birds will improve our knowledge of the evolution of these viruses, thereby preventing possible panzootic.


Assuntos
Infecções por Avulavirus/veterinária , Avulavirus/fisiologia , Galinhas , Genoma Viral , Doenças das Aves Domésticas/virologia , Sequência de Aminoácidos , Animais , Animais Selvagens , Avulavirus/genética , Infecções por Avulavirus/virologia , Doenças das Aves/virologia , Galliformes , Paquistão , Papagaios , Proteínas Virais de Fusão/análise
12.
Viruses ; 12(7)2020 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-32605292

RESUMO

Viral vectored vaccines are desirable alternatives for conventional infectious bronchitis virus (IBV) vaccines. We have recently shown that a recombinant Newcastle disease virus (rNDV) strain LaSota expressing the spike (S) protein of IBV strain Mass-41 (rLaSota/IBV-S) was a promising vaccine candidate for IBV. Here we evaluated a novel chimeric rNDV/avian paramyxovirus serotype 2 (rNDV/APMV-2) as a vaccine vector against IBV. The rNDV/APMV-2 vector was chosen because it is much safer than the rNDV strain LaSota vector, particularly for young chicks and chicken embryos. In order to determine the effectiveness of this vector, a recombinant rNDV/APMV-2 expressing the S protein of IBV strain Mass-41 (rNDV/APMV-2/IBV-S) was constructed. The protective efficacy of this vector vaccine was compared to that of the rNDV vector vaccine. In one study, groups of one-day-old specific-pathogenic-free (SPF) chickens were immunized with rLaSota/IBV-S and rNDV/APMV-2/IBV-S and challenged four weeks later with the homologous highly virulent IBV strain Mass-41. In another study, groups of broiler chickens were single (at day one or three weeks of age) or prime-boost (prime at day one and boost at three weeks of age) immunized with rLaSota/IBV-S and/or rNDV-APMV-2/IBV-S. At weeks six of age, chickens were challenged with a highly virulent IBV strain Mass-41. Our challenge study showed that novel rNDV/APMV-2/IBV-S provided similar protection as rLaSota/IBV-S in SPF chickens. However, compared to prime-boost immunization of chickens with chimeric rNDV/APMV-2, rLaSota/IBV-S and/or a live IBV vaccine, single immunization of chickens with rLaSota/IBV-S, or live IBV vaccine provided better protection against IBV. In conclusion, we have developed the novel rNDV/APMV-2 vector expressing S protein of IBV that can be a safer vaccine against IB in chickens. Our results also suggest a single immunization with a LaSota vectored IBV vaccine candidate provides better protection than prime-boost immunization regimens.


Assuntos
Avulavirus/genética , Infecções por Coronavirus/veterinária , Vetores Genéticos/genética , Vírus da Bronquite Infecciosa/imunologia , Doenças das Aves Domésticas/prevenção & controle , Vacinas Virais/administração & dosagem , Animais , Avulavirus/metabolismo , Galinhas , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/virologia , Vetores Genéticos/metabolismo , Vírus da Bronquite Infecciosa/genética , Vírus da Bronquite Infecciosa/patogenicidade , Doenças das Aves Domésticas/virologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Proteínas Virais/administração & dosagem , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/genética , Vacinas Virais/imunologia
13.
Sci Rep ; 10(1): 9532, 2020 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-32533018

RESUMO

The newly assigned subfamily Avulavirinae in the family Paramyxoviridae includes avian paramyxoviruses (APMVs) isolated from a wide variety of avian species across the globe. Till date, 21 species of APMVs are reported and their complete genome sequences are available in GenBank. The APMV genome comprises of a single stranded, negative sense, non-segmented RNA comprising six transcriptional units (except APMV-6 with seven units) each coding for a structural protein. Additionally, by co-transcriptional RNA editing of phosphoprotein (P) gene, two mRNAs coding for accessory viral proteins, V and W, are generated along with unedited P mRNA. However, in APMV-11, the unedited mRNA codes for V protein while +2 edited mRNA translates to P protein, similar to members of subfamily Rubulavirinae in the same family. Such RNA editing in paramyxoviruses enables maximizing the coding capacity of their smaller genome. The three proteins of P gene: P, V and W, share identical N terminal but varied C terminal sequences that contribute to their unique functions. Here, we analyzed the P gene editing site, V and W sequences of all 21 APMV species known so far (55 viruses) by using bioinformatics and report their genetic variations and molecular evolution. The variations observed in the sequence and hexamer phase positions of the P gene editing sites is likely to influence the levels and relative proportions of P, V and W proteins' expressions which could explain the differences in the pathogenicity of APMVs. The V protein sequences of APMVs had conserved motifs similar to V proteins of other paramyxoviruses including the seven cysteine residues involved in MDA5 interference, STAT1 degradation and interferon antagonism. Conversely, W protein sequences of APMVs were distinct. High sequence homology was observed in both V and W proteins between strains of the same species than between species except in APMV-3 which was the most divergent APMV species. The estimates of synonymous and non-synonymous substitution rates suggested negative selection pressure on the V and W proteins within species indicating their low evolution rate. The molecular clock analysis revealed higher conservation of V protein sequence compared to W protein indicating the important role played by V protein in viral replication, pathogenesis and immune evasion. However, we speculate the genetic diversity of W proteins could impact the degree of pathogenesis, variable interferon antagonistic activity and the wide host range exhibited by APMV species. Phylogenetically, V proteins of APMVs clustered into three groups similar to the recent classification of APMVs into three new genera while no such pattern could be deciphered in the analysis of W proteins except that strains of same species grouped together. This is the first comprehensive study describing in detail the genetic variations and the molecular evolution of P gene edited, accessory viral proteins of Avian paramyxoviruses.


Assuntos
Avulavirus/genética , Evolução Molecular , Variação Genética , Edição de RNA , Proteínas Virais/genética , Animais , Sequência Conservada , Genoma Viral/genética , Sinais de Exportação Nuclear , Sinais de Localização Nuclear , Filogenia , Proteínas Virais/química
14.
Vet Microbiol ; 244: 108661, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32402346

RESUMO

Pigeon paramyxovirus type 1 (PPMV-1) is an antigenic variant of avian paramyxovirus type 1, which mainly infects pigeons. Here, we characterized ten PPMV-1 viruses isolated from pigeons in China during 1996-2019. Phylogenetic analysis of available complete genomes, F and HN genes of PPMV-1 from China showed that multiple PPMV-1 genotypes (I, II, VI, and VII) exist in pigeons in China. Ten PPMV-1 viruses isolated in this study belonged to genotypes VI.1.2.2.2, VI.2.1.1.2.1, VI.2.1.1.2.2 and VII respectively. Genotype VI is predominant in pigeons. VI.2.1.1.2.2 contains most recently isolated PPMV-1 viruses, suggesting that VI.2.1.1.2.2 is a prevalent genotype in pigeons in China. In vitro and in vivo studies showed that four representative viruses from genotypes VI.2.1.1.2.1 (TA14), VI.2.1.1.2.2 (SD19), VI.1.2.2.2 (SD16), and VII (JN08) could replicate efficiently in chicken embryo fibroblasts, while the replication titer of JN08 (VII) virus was significantly lower than that of VI gene viruses in pigeon embryo fibroblasts. The TA14 (VI.2.1.1.2.1) and SD19 (VI.2.1.1.2.2) viruses caused 20 % and 30 % mortality in pigeons, respectively. No birds infected with SD16 (VI.1.2.2.2) died during the study period. JN08 (VII) virus did not cause obvious clinical signs in infected pigeons. All data indicated that VI.2.1.1.2.2 is the prevalent genotype circulating in China and poses a major threat to pigeons, suggesting that a matched vaccine is necessary to control the disease.


Assuntos
Infecções por Avulavirus/veterinária , Avulavirus/classificação , Columbidae/virologia , Genoma Viral , Filogenia , Animais , Avulavirus/isolamento & purificação , Avulavirus/patogenicidade , Infecções por Avulavirus/mortalidade , Galinhas , China , Fibroblastos/virologia , Genótipo
15.
Viruses ; 12(4)2020 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-32224965

RESUMO

Newcastle disease is an important poultry disease that also affects Columbiform birds. The viruses adapted to pigeons and doves are referred to as pigeon paramyxoviruses 1 (PPMV-1). PPMV-1 are frequently isolated from pigeons worldwide and have the potential to cause disease in chickens. The complete genomes of 18 PPMV-1 isolated in China during 2012-2018 were sequenced by next-generation sequencing (NGS). Comprehensive phylogenetic analyses showed that five of the viruses belong to sub-genotype VI1.2.1.1.2.1 and 13 isolates belong to sub-genotype VI.2.1.1.2.2. The results demonstrate that these sub-genotypes have been predominant in China during the last decade. The viruses of these sub-genotypes have been independently maintained and continuously evolved for over 20 years, and differ significantly from those causing outbreaks worldwide during the 1980s to 2010s. The viral reservoir remains unknown and possibilities of the viruses being maintained in both pigeon farms and wild bird populations are viable. In vivo characterization of the isolates' pathogenicity estimated mean death times between 62 and 114 hours and intracerebral pathogenicity indices between 0.00 and 0.63. Cross-reactivity testing showed minor antigenic differences between the studied viruses and the genotype II LaSota vaccine. These data will facilitate PPMV-1 epidemiology studies, vaccine development, and control of Newcastle disease in pigeons and poultry.


Assuntos
Infecções por Avulavirus/veterinária , Avulavirus/genética , Columbidae/virologia , Genoma Viral , Genômica , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/virologia , Animais , Avulavirus/imunologia , Avulavirus/isolamento & purificação , China/epidemiologia , Reações Cruzadas , Genômica/métodos , Genótipo , História do Século XXI , Epidemiologia Molecular , Filogenia , Doenças das Aves Domésticas/história , Doenças das Aves Domésticas/imunologia , Sequenciamento Completo do Genoma
16.
Sci Rep ; 10(1): 2221, 2020 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-32042001

RESUMO

Highly pathogenic avian influenza (HPAI) is a devastating disease of poultry and a serious threat to public health. Vaccination with inactivated virus vaccines has been applied for several years as one of the major policies to control highly pathogenic avian influenza virus (HPAIV) infections in chickens. Viral-vectored HA protein vaccines are a desirable alternative for inactivated vaccines. However, each viral vector possesses its own advantages and disadvantages for the development of a HA-based vaccine against HPAIV. Recombinant Newcastle disease virus (rNDV) strain LaSota expressing HA protein vaccine has shown promising results against HPAIV; however, its replication is restricted only to the respiratory tract. Therefore, we thought to evaluate avian paramyxovirus serotype 3 (APMV-3) strain Netherlands as a safe vaccine vector against HPAIV, which has high efficiency replication in a greater range of host organs. In this study, we generated rAPMV-3 expressing the HA protein of H5N1 HPAIV using reverse genetics and evaluated the induction of neutralizing antibodies and protection by rAPMV3 and rNDV expressing the HA protein against HPAIV challenge in chickens. Our results showed that immunization of chickens with rAPMV-3 or rNDV expressing HA protein provided complete protection against HPAIV challenge. However, immunization of chickens with rAPMV-3 expressing HA protein induced higher level of neutralizing antibodies compared to that of rNDV expressing HA protein. These results suggest that a rAPMV-3 expressing HA protein might be a better vaccine for mass-vaccination of commercial chickens in field conditions.


Assuntos
Avulavirus/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Virus da Influenza A Subtipo H5N1/patogenicidade , Vacinas contra Influenza/imunologia , Influenza Aviária/prevenção & controle , Criação de Animais Domésticos , Animais , Anticorpos Neutralizantes/análise , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/análise , Anticorpos Antivirais/imunologia , Avulavirus/genética , Galinhas , Vetores Genéticos/genética , Vetores Genéticos/imunologia , Imunogenicidade da Vacina , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Influenza Aviária/virologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
17.
J Gen Virol ; 101(2): 156-167, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31922948

RESUMO

Several pigeon paramyxovirus-1 (PPMV-1) outbreaks in feral pigeons were described recently in Switzerland. The potential of PPMV-1 to induce the notifiable Newcastle disease in chickens is discussed controversially. Therefore, in order to study epidemiologically relevant parameters such as the kinetics of PPMV-1 replication and shedding as well as seroconversion after infection, chickens were infected experimentally with a Swiss PPMV-1 isolate. This generated also defined sample material for the comparison of diagnostic tests. The infectivity of the Swiss PPMV-1 isolate for chickens was demonstrated successfully by virus shedding after experimental inoculation. Our data suggest that long-lasting shedding for up to 60 days can occur in chickens infected with PPMV-1. The isolate used here was of low pathogenicity for chickens. Different quantitative reverse transcription PCR assays were evaluated with a set of Swiss PPMV-1 isolates, and various samples from experimentally infected chickens were analysed with respect to their suitability for viral RNA detection. At 14 days post-infection, virus genome was detected mainly in spleen, caecal tonsils, heart, cloacal swabs, liver, proventriculus, duodenum and kidney tissue samples. Overall, the level of virus replication was low. Not all assays used routinely in diagnostics were capable of detecting viral genome from the isolates tested. Possible explanations are the genetic divergence of PPMV-1 and the low level of viral RNA in the samples. In contrast, two methods that are not used routinely proved more suitable for virus-genome detection. Importantly, the collection of material from various different organs is recommended, in addition to the kidney and brain analysed routinely. In conclusion, this study shows that there is a need to reconsider the type of samples and the protocols used for the detection of PPMV-1 RNA in chickens.


Assuntos
Infecções por Avulavirus/diagnóstico , Avulavirus , Doença de Newcastle/diagnóstico , Animais , Avulavirus/genética , Avulavirus/crescimento & desenvolvimento , Avulavirus/isolamento & purificação , Avulavirus/patogenicidade , Infecções por Avulavirus/patologia , Galinhas , Columbidae/virologia , Genoma Viral , Doença de Newcastle/patologia , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/crescimento & desenvolvimento , Vírus da Doença de Newcastle/isolamento & purificação , Vírus da Doença de Newcastle/patogenicidade , Doenças das Aves Domésticas/virologia , Suíça , Viroses/veterinária , Replicação Viral , Eliminação de Partículas Virais
18.
Vet Microbiol ; 236: 108377, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31500723

RESUMO

Avian paramyxoviruses (APMVs) consist of twenty known species and have been isolated from domestic and wild birds around the world. In 2009, the isolate APMV/dove/Taiwan/AHRI33/2009 was isolated from swabs of red turtle doves (Streptopelia tranquebarica) during active surveillance of avian influenza in resident birds in Taiwan, and it was initially identified as paramyxovirus based on electron microscopy. Hemagglutination inhibition assays indicated antigenic heterogeneity of AHRI33 with the known APMV-1, -2, -3, -4, -6, -8, and -9 species, only showing weak but measurable cross-reactivity with APMV-7. Pathogenicity ICPI test revealed that the virus was avirulent for chickens. The AHRI33 virus genome revealed a typical APMV structure consisting of six genes 3'-NP-P-M-F-HN-L-5', and the length of the genome was 16,914 nucleotides, the third longest among the members of the subfamily Avulavirinae. Estimates of the nucleotide sequence identities of the genome between each prototype of APMVs had shown AHRI33 to be more closely related to APMV-7 than to the others, with a sequence identity of 62.8%. Based on topology of the phylogenetic tree of RdRp genes and the branch length between the nearest node and the tip of the branch, AHRI33 met the criteria for designation as distinct species. Together, the data suggest that the isolate APMV/dove/Taiwan/AHRI33/2009 should be considered as the prototype strain of the new species Avian metaavulavirus 21 in the genus Metaavulavirus in the subfamily Avulavirinae.


Assuntos
Avulavirus/isolamento & purificação , Columbidae/virologia , Sequência de Aminoácidos , Animais , Avulavirus/genética , Regulação Viral da Expressão Gênica , Variação Genética , Genoma Viral , Filogenia , Proteínas Virais/genética , Proteínas Virais/metabolismo
19.
Avian Pathol ; 48(6): 610-621, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31403322

RESUMO

Newcastle disease (ND), caused by virulent Avian avulavirus 1 (AAvV 1), affects a wide range of avian species worldwide. Recently, several AAvVs of diverse genotypes have emerged with varying genomic and residue substitutions, and subsequent clinical impact on susceptible avian species. We assessed the clinico-pathological influence of two different AAvV 1 pathotypes [wild bird originated-velogenic strain (sub-genotype VIIi, MF437287) and feral pigeon originated-mesogenic strain (sub-genotype VIm, KU885949)] in commercial broiler chickens and pigeons. The velogenic strain caused 100% mortality in both avian species while the mesogenic strain caused 0% and 30% mortality in chickens and pigeons, respectively. Both strains showed tissue tropism for multiple tissues including visceral organs; however, minor variances were observed according to host and pathotype. The observed gross and microscopic lesions were typical of AAvV 1 infection. Utilizing oropharyngeal and cloacal swabs, a comparable pattern of viral shedding was observed for both strains from each of the infected individuals of both avian species. The study concludes a varying susceptibility of chickens and pigeons to different wild bird-originated AAvV 1 pathotypes and, therefore, suggests continuous monitoring and surveillance of currently prevailing strains for effective control of the disease worldwide, particularly in disease-endemic countries.


Assuntos
Infecções por Avulavirus/veterinária , Avulavirus/genética , Doenças das Aves/patologia , Galinhas/virologia , Columbidae/virologia , Doença de Newcastle/patologia , Doenças das Aves Domésticas/patologia , Animais , Avulavirus/isolamento & purificação , Infecções por Avulavirus/patologia , Infecções por Avulavirus/virologia , Doenças das Aves/virologia , Genômica , Genótipo , Doença de Newcastle/virologia , Doenças das Aves Domésticas/virologia , Taxa de Sobrevida
20.
Viruses ; 11(7)2019 07 13.
Artigo em Inglês | MEDLINE | ID: mdl-31337066

RESUMO

Avian orthoavulavirus 13 (AOAV-13), also named avian paramyxovirus 13 (APMV-13), has been found sporadically in wild birds around the world ever since the discovery of AOAV-13 (AOAV-13/wild goose/Shimane/67/2000) in a wild goose from Japan in 2000. However, there are no reports of AOAV-13 in China. In the present study, a novel AOAV-13 virus (AOAV-13/wild goose/China/Hubei/V93-1/2015), isolated from a wild migratory waterfowl in a wetland of Hubei province of China, during active surveillance from 2013 to 2018, was biologically and genetically characterized. Phylogenetic analyses demonstrated a very close genetic relationship among all AOAV-13 strains, as revealed by very few genetic variations. Moreover, pathogenicity tests indicated that the V93-1 strain is a low virulent virus for chickens. However, the genome of the V93-1 virus was found to be 16,158 nucleotides (nt) in length, which is 12 nt or 162 nt longer than the other AOAV-13 strains that have been reported to date. The length difference of 12 nt in strain V93-1 is due to the existence of three repeats of the conserved sequence, "AAAAAT", in the 5'-end trailer of the genome. Moreover, the HN gene of the V93-1 virus is 2070 nt in size, encoding 610 aa, which is the same size as the AOAV-13 strain from Japan, whereas that of two strains from Ukraine and Kazakhstan are 2080 nt in length, encoding 579 aa. We describe a novel AOAV-13 in migratory waterfowl in China, which suggests that diversified trailer region sequences and HN gene lengths exist within serotype AOAV-13, and highlight the need for its constant surveillance in poultry from live animal markets, and especially migratory birds.


Assuntos
Animais Selvagens/virologia , Infecções por Avulavirus/veterinária , Avulavirus/classificação , Genoma Viral , Proteína HN/genética , Migração Animal , Animais , Avulavirus/isolamento & purificação , Galinhas/virologia , China , Patos/virologia , Gansos/virologia , Filogenia , RNA Viral/genética , Análise de Sequência de DNA , Sorogrupo
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