Resumo
The aim of the present study was to evaluate the post-vaccinal reaction to two lentogenic vaccine strains of Newcatle disease virus (NDV) and a recombinant turkey herpesvirus (rHVT) vaccine expressing the fusion glycoprotein of NDV in broiler chickens through histomorphometric and histopathologic analyses of the trachea. The experiment involved 245 chicks housed in randomized blocks with three different enclosures under controlled conditions of temperature, light and ventilation. Each enclosure represented a vaccine strain and was divided into groups according to the administration route. Each block also had its own control group composed of unvaccinated birds. The vaccine strains PHY.LMV.42 (PL42) and La Sota (LS) were selected according to the Intracerebral Pathogenicity Index (ICPI) and the rHVT-NDV Serotype 3 strain (ST3) was selected for representing non-NDV infection. At two, four, seven, 14 and 21 days post vaccination, fragments from the middle third of the trachea were collected and submitted to routine histological processing. For the histomorphometric analysis, the slides were photographed, and the thickness of the tracheal mucosa was measured. Statistical analysis involved two-way ANOVA and Tukey's post-hoc test with a 5% significance level. For the histopathological evaluation, lesions were described as to the degree of intensity and distribution. At four and 14 days post vaccination with the LS strain administered by the ocular route, the means of thickening of the tracheal mucosa (20.85±7.31µm and 26.97±5.50µm, respectively) were significantly higher (p<0.05) than for all other strains, which was related to the severe histopathological lesions found in this group, characterized by hyperemia, hyperplasia of the mucous glands, moderate deciliation and multifocal lymphohistiocytic inflammatory infiltrate. At 21 days, broiler chickens vaccinated with the ST3 strain showed more discrete lesions and less thickening of the tracheal mucosa (23.23±7.62µm; p<0.05) in comparison with other studied strains. The lesions found in this group were only hemorrhage, deciliation and mild focal lymphocytic inflammatory infiltrate. The results of the histomorphometry and histopathology of the trachea indicated that vaccination with rHVT-NDV Serotype 3 strain induced lower degree post-vaccine tracheal lesions compared to other vaccine strains analyzed in this study.
Objetivou-se avaliar a reação pós-vacinal de duas estirpes lentogênicas do vírus da doença de Newcastle (VDN) e uma vacina recombinante de herpesvirus de perus (rHVT) que expressa a glicoproteína de fusão de VDN em frangos de corte por meio da histomorfometria e histopatologia da traqueia. Foram utilizados 245 pintos alojados em blocos ao acaso, sendo três galpões distintos em condições controladas de temperatura, luz e ventilação. Cada galpão representou uma cepa vacinal, onde foram divididos por grupos de acordo com a via de administração. Todos os blocos possuíam um grupo controle composto por aves não vacinadas. As cepas vacinais PHY.LMV.42 (PL42) e La Sota (LS) utilizadas foram selecionadas de acordo com o Índice de Patogenicidade Intracerebral (IPIC) e a cepa Sorotipo 3 (ST3), da vacina rHVT-VDN foi selecionada por não representar infecção do VDN. Aos dois, quarto, sete, 14 e 21 dias pós-vacinação, fragmentos do terço médio da traqueia foram coletados e posteriormente processados conforme rotina histológica. Para análise histomorfométrica da mucosa traqueal, as lâminas foram fotografadas e realizadas as mensurações da espessura da mucosa traqueal sendo aplicado teste de anaÌlise de variaÌncia a dois criteÌrios (ANOVA) e utilizando o post-hoc de Tukey com niÌvel de significaÌncia de 5%. Para a avaliação histopatológica foram observadas a presença de lesões microscópicas e estas foram descritas quanto ao grau de intensidade e distribuição. Aos quatro e quatorze dias pós-vacinação com a cepa LS administrada por via ocular, as médias do espessamento da mucosa traqueal (20,85±7,31µm e 26,97±5,50µm, respectivamente) foram significativamente maiores (p<0,05) quando comparada a todas as demais cepas utilizadas, isto se deve às severas lesões histopatológicas encontrados neste grupo, caracterizadas por hiperemia, hiperplasia das glândulas mucosas, deciliação moderada e infiltrado inflamatório linfohistiocitário multifocal moderado. Já aos 21 dias as aves vacinadas com a cepa ST3 apresentaram lesões mais discretas e menor espessamento da mucosa da traqueia (23,23±7,62µm; p<0,05) em comparação às demais cepas estudadas. As lesões encontradas neste grupo foram apenas hemorragia, deciliação e infiltrado inflamatório linfocitário focal discreto. Os resultados da histomorfometria e da histopatologia da traqueia indicou que a vacinação com a rHVT-NDV, cepa Sorotipo 3 induziu menor grau de lesões pós-vacinais na traqueia comparada a outras cepas vacinais analisadas nesse estudo.
Assuntos
Animais , Traqueia/efeitos dos fármacos , Traqueia/patologia , Vírus da Doença de Newcastle/imunologia , Vacinas/efeitos adversos , Galinhas/virologia , Doença de Newcastle/imunologia , Vacinação/efeitos adversosResumo
ABSTRACT: Chickens are considered to be potential reservoirs of Newcastle disease virus (NDV). In this study, six Newcastle disease virus strains were isolated and characterized in Tibetan chickens. The HN gene was sequenced, and phylogenetic relationship to reference strains was studied. The phylogenetic analysis demonstrated that these six isolated strains were closely related to NDV isolates of the reference strains GQ245823, KT002186, KU527561, KJ563939, AY225110, EU305607, KM056357, Y18898, GQ245832, AF077761 and lasota strain. Among them, EU305607, KJ563939 and KM056357 were isolated from India, while lasota strain came from attenuated vaccine widely used in China. Then, mean death time (MDT) and intracerebral pathogenicity index (ICPI) were used to estimate the pathogenicity of the isolates. Pathogenicity experiment showed HNH1 and HN17 to be virulent. Our results indicated that genetically diverse viruses circulate in Tibetan chickens, and based upon the phlogeographic analysis, we estimated the origin of ancestral viruses of the isolates and its sister strains located in India and China (lasota strain). It indicates the importance of continuous surveillance to enhance current understanding of the genetic evolution of the NDV strains.
Resumo
Chickens are considered to be potential reservoirs of Newcastle disease virus (NDV). In this study, six Newcastle disease virus strains were isolated and characterized in Tibetan chickens. The HN gene was sequenced, and phylogenetic relationship to reference strains was studied. The phylogenetic analysis demonstrated that these six isolated strains were closely related to NDV isolates of the reference strains GQ245823, KT002186, KU527561, KJ563939, AY225110, EU305607, KM056357, Y18898, GQ245832, AF077761 and lasota strain. Among them, EU305607, KJ563939 and KM056357 were isolated from India, while lasota strain came from attenuated vaccine widely used in China. Then, mean death time (MDT) and intracerebral pathogenicity index (ICPI) were used to estimate the pathogenicity of the isolates. Pathogenicity experiment showed HNH1 and HN17 to be virulent. Our results indicated that genetically diverse viruses circulate in Tibetan chickens, and based upon the phlogeographic analysis, we estimated the origin of ancestral viruses of the isolates and its sister strains located in India and China (lasota strain). It indicates the importance of continuous surveillance to enhance current understanding of the genetic evolution of the NDV strains.(AU)
Assuntos
Animais , Feminino , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/patogenicidade , Galinhas/virologia , Filogenia , TibetResumo
Chickens are considered to be potential reservoirs of Newcastle disease virus (NDV). In this study, six Newcastle disease virus strains were isolated and characterized in Tibetan chickens. The HN gene was sequenced, and phylogenetic relationship to reference strains was studied. The phylogenetic analysis demonstrated that these six isolated strains were closely related to NDV isolates of the reference strains GQ245823, KT002186, KU527561, KJ563939, AY225110, EU305607, KM056357, Y18898, GQ245832, AF077761 and lasota strain. Among them, EU305607, KJ563939 and KM056357 were isolated from India, while lasota strain came from attenuated vaccine widely used in China. Then, mean death time (MDT) and intracerebral pathogenicity index (ICPI) were used to estimate the pathogenicity of the isolates. Pathogenicity experiment showed HNH1 and HN17 to be virulent. Our results indicated that genetically diverse viruses circulate in Tibetan chickens, and based upon the phlogeographic analysis, we estimated the origin of ancestral viruses of the isolates and its sister strains located in India and China (lasota strain). It indicates the importance of continuous surveillance to enhance current understanding of the genetic evolution of the NDV strains.(AU)
Assuntos
Animais , Feminino , Galinhas/virologia , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/patogenicidade , Filogenia , TibetResumo
This study was carried out during 2002/2003, aiming to determine the prevalence of virulent Newcastle disease virus strains (NDV) in Brazilian commercial poultry farms. Clinical samples were obtained from the Southeastern, Southern and Central-Western regions, which comprise the main area of the Brazilian poultry production. Serum samples and tracheal and cloacal swabs of 23,745 broiler chickens from 1,583 flocks, including both vaccinated chickens and those with no vaccination information, were tested for NDV using a diagnostic ELISA kit. The seropositivity was 39.1%, and the isolation percentage by flock varied from 1.0 to 7.6%, and by region from 6.5 to 58.4%. Higher isolation rates (74.3-83.3%) were obtained after three passages in embryonated chicken eggs. All isolates preliminarily identified as NDV were characterized as nonpathogenic strains, as their Intracerebral Pathogenicity Index (ICPI) was below 0.7. Based on results of this study, Brazil can claim a virulent NDV-free status for commercial flocks.
Resumo
Intracerebral pathogenicity index (ICPI) and mean death time (MDT) were determined using commercial live vaccines against Newcastle disease available in Brazil. The ICPI profiles obtained for B1 vaccine strains were nonvirulent and varied from 0 to 0.19, and their MDT was 104-116 hours. The LaSota strains had an ICPI varying between 0.02 and 0.37 and MDT from 92 to 116 hours. ICPI and MDT for the Clone 30 were 0.11 and 104 hours, respectively. For Ulster vaccines, ICPI and MDT were 0 and >150 hours; for VG-GA was 0.03 and 140 hours; and for C2, 0.04 and >144 hours. Eye drop vaccination and IM challenge, at the 1st week and the 4th week, respectively, resulted in highest protection for B1 (95-100%) and LaSota (90-100%) strains. The variability in vaccine ICPI did not interfere with immune response and all vaccines provided similar protection. All vaccines were considered non virulent and were classified as lentogenic according to the immunobiological product standards.
Resumo
Intracerebral pathogenicity index (ICPI) and mean death time (MDT) were determined using commercial live vaccines against Newcastle disease available in Brazil. The ICPI profiles obtained for B1 vaccine strains were nonvirulent and varied from 0 to 0.19, and their MDT was 104-116 hours. The LaSota strains had an ICPI varying between 0.02 and 0.37 and MDT from 92 to 116 hours. ICPI and MDT for the Clone 30 were 0.11 and 104 hours, respectively. For Ulster vaccines, ICPI and MDT were 0 and >150 hours; for VG-GA was 0.03 and 140 hours; and for C2, 0.04 and >144 hours. Eye drop vaccination and IM challenge, at the 1st week and the 4th week, respectively, resulted in highest protection for B1 (95-100%) and LaSota (90-100%) strains. The variability in vaccine ICPI did not interfere with immune response and all vaccines provided similar protection. All vaccines were considered non virulent and were classified as lentogenic according to the immunobiological product standards.