The effect of parental immunity on the response of five-days-old chicks to intranasal vaccination with F strain Newcastle disease virus

Summary Two flocks of chicks from parents not immune to Newcastle disease (ND) virus and six flocks from immune parents were vaccinated intranasally with 10^6.03 to 10^6.86 mean embryo infective doses of F strain ND virus on the fifth day of life. A group of chicks from each flock was left unvaccinated. Vaccinated and unvaccinated chicks from each flock were killed on days one to ten and then every other day until 39 to 61 days. Serums were examined by the ND virus haemagglutination-inhibition (HI) test, and tissues were inoculated into fertile hens' eggs for recovery of vaccine virus. Vaccine virus was recovered from vaccinated nonparentally-immune chicks daily from two to 14 days post-vaccination. Their serological response was detectable two days post-vaccination and reached peak levels nine to 14 days post-vaccination. In contrast, vaccine virus was recovered rarely or not at all from parentally-immune chicks and their serological response was much less than in chicks with no parental immunity. It is concluded that parentally-acquired antibodies neutralised vaccine virus so reducing the amount of immunizing virus antigen to which the host was exposed.

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Sumario Dos lotes de pollos de padres no inmunes a la enfermedad de Newcastle y seis lotes de pollos de padres inmunes fueron vacunados instranasalmente con 10^6.08 a 10^6.86 dosis infectantes medias para embriones de la cepa F del virus de Newcastle, el dia quinto despues del nacimiento. Un grupo de pollitos de cada lote fue dejado sin vacunar. Pollos vacunados y no vacunados de cada lote fueron sacrificados desde el dia primero hasta el dia décimo despues de la vacunación y luego dejando un dia hasta los dias 39 a 61. Los sueros fueron examinados por la prueba de la inhibición de la hemaglutinación del virus de Newcastle y los tejidos fueron inoculados en huevos fértiles de gallina con el fin de recobrar el virus vacunal. El virus vacunal fue recobrado de pollitos vacunados sin inmunidad parental desde el dia dos hasta el dia 14 despues de la vacunación. La respuesta serológica de los pollos fue detectable dos dias despues de la vacunación y alcanzó los niveles mas altos del noveno al catorceavo dia. En contraste, el virus vacunal se recobró muy raramente ó no se recuperó de pollos con immunidad parental y sus respuestas serológicas fue mucho menor que en pollitos sin inmunidad parental. Se concluye que los anticuerpos adquiridos pasivamente neutralizaron al virus vacunal reduciendo así la cantidad de antígeno viral inmunizante a la que el huesped se había expuesto.

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Résumé 2 bandes de poussins issues de parents non immuns au virus de la maladie de Newcastle et 6 autres bandes issues de parents immuns furent vaccinés par la voie nasale au 5ème jour de leur vie avec 10^6.08 à 10^6.086 doses moyennes infectantes pour l'embryon de la souche F du virus de Newcastle. Dans chaque bande, un groupe de poussins fut laissé non vacciné. Des poussins de chaque bande, vaccinés et non vaccinés, furent sacrifiés chaque jour, du jour 1 au jour 10 et ensuite du jour 39 jusqu'au jour 61. Leurs sérums furent examinés au moyen du test d'inhibition de l'hémagglutination et leurs tissus furent inoculés à des oeufs de poules embryonnés pour essayer de retrouver le virus vaccinal. Celui-ci fut décelé du 1er au 14ème jour après la vaccination chez les poussins vaccinés et issus de parents non immuns. Leur réponse sérologique était décelable 2 jours après la vaccination et atteignait son niveau maximum du 9ème au 14ème jour après cette vaccination. Au contraire le virus vaccinal n'était retrouvé que rarement ou pas du tout chez les poussins à immunité parentale et la réponse sérologique de ces derniers était bien moindre que celle des poussins sans immunité transmise. On en conclut que les anticorps d'origine parentale neutralisent le virus vaccinal, réduisant ainsi la quantité d'antigène viral immunisant par laquelle l'organisme est sollicité.

     

A simple method for the isolation of immunoglobulin (Y) from the eggs of immunized hens

Hens were vaccinated with Salmonella typhi-murium and with serum-IgG from a goat. Antibodies (IgY) were isolated from the eggs by ammonium sulphate precipitation. The concentration of egg antibodies was equal or superior to the concentration of serum antibodies of the hen. The longlasting antibody plateau of 9 and 28 weeks, respectively, provides evidence of the high yields of specific IgY which can be collected from a single immunized hen.     

Effect of sample selection from experimentally contaminated hatching eggs and freshly hatched chicks on Salmonella enteritidis detection rate.

The conventional culture methods of hatching eggs using shell and/or egg contents for detection of salmonella organisms give mostly unsatisfactory results. The aim of the present study is to evaluate selection of other samples and techniques of culturing hatching eggs and freshly hatched chicks. This current study provides the best evidence of Salmonella enteritidis in artificially contaminated eggs (Layer type) by using enrichment broth in empty egg shell samples in comparison to culturing samples from yolk, albumen or from shell above the air cell (with the outer shell membrane). The isolation rates could be enhanced if empty egg shell was initially filled with Buffered Pepton Water as pre-enrichment broth. Examination of organs from freshly hatched chicks revealed that crop samples give mostly higher reisolation rates.     

鸡贫血病—法氏囊病疫苗免疫母鸡后子代雏鸡的免疫学变化

本项目应用现代免疫学新技术对鸡传染性贫血病（CIA）-传染性法氏囊病（IBD）疫苗联合免疫母鸡后,其子代雏鸡外周血液T、B细胞数量和IgG、IgM、IgA含量法及法氏囊、胸腺、脾脏、盲肠扁桃体、哈德尔腺的T细胞和IgG、IgM、IgA抗体生成细胞数量以及泪液、气管液、胆汁、肠液的IgA、IgM、IgG含量的变化进行了动态研究。结果发现,CIA-IBD疫苗联合免疫母鸡后,其子代雏鸡外周血液、免疫器官组织和局部体液的上述各项指标均不同程度地高于未免疫的相应对照雏鸡。表明CIA-IBD疫苗免疫母鸡后,其子代雏鸡的体液免疫和细胞免疫功能明显增强,而CIAV-IBDV强毒攻击后,未免疫的子代雏鸡,其外周血液,免疫器官组织和局部体液的各项免疫学指标均明显低于疫苗免疫攻毒的子代雏鸡,这与未免疫雏鸡缺乏特异性抗体,强毒攻击后,雏鸡免疫器官组织广泛损害,淋巴细胞变性坏死等有关。     

Ultrastructural changes of the tracheal epithelium after vaccination of day-old chickens with the La Sota strain of Newcastle disease virus

The progression of tracheal lesions induced by vaccination of day-old specific pathogen-free chicks with the La Sota strain of Newcastle disease virus (NDV) was examined by relating surface changes as observed by scanning electron microscopy with subcellular changes seen by transmission electron microscopy. NDV infection resulted in hypertrophy of goblet cells, their rupture, and the formation of excess mucus. Activation of goblet cells peaked within 4 days postvaccination. Afterward, the activation levels gradually decreased. At the level of the ciliated cells, a marked increase in the proportion of nonciliated to ciliated cells and later an almost complete deciliation of the tracheal surface were observed because a simple squamous to cuboidal epithelium replaced the original pseudostratified epithelium. Fifteen days postvaccination, all epithelial damage was restored. Because the observed vaccination-induced lesions are detrimental to epithelial integrity and function as a barrier against invading microorganisms, they might explain at the ultrastructural level the secondary complications of vaccination with the La Sota strain against NDV.     

从种蛋中分离的新城疫病毒抗原性及其F和HN基因的分析

对3株首次从种蛋中分离到的NDV进行交叉HI试验表明,这3株分离毒的交叉HI同源性为98.1%～100%。对这3株病毒及另1株从输卵管分离到的NDV的F与HN完整基因进行扩增并克隆测序,序列结果登陆GenBank(FJ011441～FJ011448)。氨基酸同源性分析表明:这4个毒株的F与HN基因同源性分别为99.5%～99.8%和99.6%～100%,远高于其与Lasota、F48E8株以及目前国内流行的其他基因Ⅶ型毒株的同源性。分离株F基因裂解位点的氨基酸序列为112R-R-Q-K-R-F117,符合NDV强毒株特征,根据绘制的遗传进化树分析,属于基因Ⅶ型。研究结果显示这4个不同来源毒株其2个基因的同步高度同源性可能与它们均是与生殖道感染有关。     

免疫鸭体内副黏病毒的分离及其F基因的分子特征

为分析当地鸭副黏病毒流行状况及其融合蛋白(F)基因的序列特征,从江苏省某新城疫疫苗免疫过的鸭群中分离到1株病毒,经血清学试验和中和试验鉴定为鸭副黏病毒,命名为JS0920株.生物学特性分析,9日龄SPF鸡胚的半数致死量(ELD50)为10-5.3；9日龄SPF鸡胚的最小致死量平均死亡时间(MDT)为47.7h,1日龄SPF鸡脑内接种致病指数(ICPI)为1.875,6周龄鸡静脉接种致病指数(IVPI)测定为2.7.用该病毒的培养物(鸡胚尿囊液)经肌肉注射攻毒,鸭的发病率为70％,死亡率为40％；鸡的发病率和死亡率均为100％.用RT-PCR方法扩增该分离毒株的F基因,序列分析表明,JS0920株与标准强毒株F48E9的核苷酸序列和氨基酸序列同源性最高,分别为99.2％和99.5％,与免疫预防用的LaSota株的同源性分别为89.0％和92.2％；其多肽裂解位点为112R-R-Q-R-R-F117,具有高致病性鸭副黏病毒毒株的分子特征；系统发育进化树分析,JS0920株与LaSota株的基因型不同,JS0920株为基因Ⅸ型,与F48E9株的亲缘关系最近.试验结果对认识当前DPMV的流行和变异现状以及疫苗研制具有重要的参考价值.     

Susceptibility of chicks to infectious bursal disease (IBD) following vaccination of their parents with live IBD vaccine.

Vaccination of parent chickens with a commercial live infectious bursal disease (IBD) vaccine under field conditions at varying ages and by different routes resulted in variable susceptibility to the disease in their chicks. There was little correlation between the methods of vaccination and the levels of immunity in the chicks. There was some evidence that levels of transferred immunity decreased with the age of the parents. Of five flocks examined, the onset of susceptibility to IBD occurred at two days of age in two flocks and about one week of age in the other three. Four flocks were completely susceptible by the 27th day, the other at 31 days. Precipitins were demonstrated at day old in 100 per cent of chicks in three flocks and in 80 per cent in the other two. The precipitins had disappeared in all flocks by 17 days of age.     

一株野鸭源新城疫病毒F基因的克隆及序列分析

根据GenBank中新城疫病毒(Newcastle disease virus,NDV)的F基因序列设计了1对特异性引物,成功地对其中一株野鸭源NDV的F基因进行了克隆、测序,并推导其氨基酸序列,选择F基因部分片段绘制了遗传进化树。F基因全长1 662 bp,单一的阅读框架编码553个氨基酸,构成的F蛋白上有13个Cys残基位点和6个潜在的糖基化位点,其裂解位点的氨基酸序列为112R-R-Q-K-116R-117F。遗传进化分析表明,该NDV分离株为基因Ⅶ型NDV,与近年来我国家禽中所报道的NDV的基因型相一致。     

Oral vaccination of chickens with the V4 strain of Newcastle disease virus. Cooked and raw white rice as a vehicle

Summary Uncooked white rice and cooked white rice were tested as vehicles for the V4 strain of oral Newcastle disease vaccine. The results of feeding experiments were evaluated by the measurement of haemagglutination inhibition antibodies against Newcastle disease virus. Little of the virus applied to uncooked white rice could be recovered, even immediately after mixing, whereas when the virus was applied to cooked white rice most of it could be recovered. In 4 separate experiments, chickens failed to respond serologically to vaccine supplied on uncooked white rice. In all of 4 experiments with cooked white rice, there were serological responses in vaccinated chickens, from 45% to 100% of the chickens developing titres sufficiently high to indicate protection against challenge with virulent virus. Development of haemagglutination inhibition antibodies in some control chickens indicated the ability of the vaccine virus for lateral spread or persistence in the environment.

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Vacunacion Oral De Pollos Con La Cepa V4 Del Virus De La Enfermedad De Newcastle Utilizando Como Vehiculo Arroz Blanco Cocinado Y No Cocinado

Resumen Se utilizo arroz blanco sin cocinar y cocinado como vehículo para la vacuna oral con la cepa V4 de la enfermedad de Newcastle. Los resultados de los experimentos fueron evaluados mediante la determinación de los anticuerpos de inhibición-hemaglutinación contra el virus de la enfermedad de Newcastle. Poca cantidad del virus de la enfermedad de Newcastle pudo recuperarse en el arroz crudo, incluso inmediatamente despues de revolverlo, mientras que la mayoría del virus pudo recuperarse del arroz cocinado. En 4 experimentos separados, los pollos no respondieron serológicamente a la vacuna aplicada en arroz blanco crudo. En los 4 experimentos con arroz cocinado, se detectaron respuestas serológicas en los pollos vacunados; 45% a 100% de los pollos desarrollaron títulos suficientes como para resistir la descarga de virus virulento. El desarrollo de anticuerpos de inhibición-hemaglutinación en algunos de los pollos controles indicó la capacidad del virus vacunal para dispersarse lateralmente y persistir en el ambiente.

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Vaccination Par Voie Orale De Poulets Avec La Souche V4 Du Virus De La Maladie De Newcastle Avec Comme Support Du Riz Blanc Cuit Et Du Riz Blanc Cru

Résumé Du riz blanc crû et du riz blanc cuit ont été testés en tant que supports de la souche de virus V4 du vaccin contre la maladie de Newcastle. Les résultats de ces expériences d'alimentation ont été évalués par la mesure des anticorps d'inhibition de l'hémagglutination du virus de la maladie de Newcastle.Seule une petite quantité du virus mélangé dans le riz blanc crû a pu être récupérée même immédiatement après le mélange. Par contre la majeur partie du virus a pu être récupérée lorsqu'il s'agissait du riz blanc cuit.Dans 4 expériences séparées, ils n'y avait pas de réponses sérologiques chez les poulets recevant du vaccin ajouté au riz blanc crû. Dans toutes les 4 expériences avec du riz blanc cuit, les réponses sérologiques ont été positives chez les poulets vaccines, 45p. 100 à 100p. 100 des poulets développant des titres suffisamment élevés pour démontrer une protection contre l'épreuve avec du virus virulent. Le développement des anticorps d'inhibition de l'hémagglutination chez quelques poulets témoins a montré la capacité de diffusion latérale ou la persistance dans l'environnement du virus.

     

Protective antibody response following oral vaccination of feral pigeons (<Emphasis Type="Italic">Columba livia</Emphasis>) with Newcastle disease vaccine (strain I-2) coated on oiled rice

The novel vaccination technique for feral pigeons was developed in the present study. Multi-age feral pigeons were vaccinated orally with Newcastle disease (ND) strain I-2 vaccine coated on oiled rice. The results showed that 14 days after vaccination 40% of pigeons seroconverted with HI GMT of ≥3 log₂ whereas 28 days after vaccination the seroconversion rate of these birds reached 100%. Moreover, all vaccinated pigeons survived the challenge of virulent Newcastle disease virus (NDV). The findings from the present study indicated that the use of ND (strain I-2) vaccine in feral pigeons is feasible and resulted into the production of protective antibody response. Thus ND I-2 vaccine may prevent the spread of NDV to other birds particularly chickens. Furthermore the use of oral vaccine in feral multi-age pigeons overcomes the difficulty of catching these birds for individual vaccination.     

嗜水气单胞菌免疫蛋鸡IgY效价变化规律和体外抑菌效果观察

大约在100多年以前,Klemperer发现免疫的鸡可以将Ig从血清转移到蛋黄中,具有IgG样的免疫球蛋白作用.用禽卵大量制备多克隆抗体它有几个优势：饲养成本较低;不会造成动物的伤害;易产生高水平的抗体;在pH4～11时较稳定,可作为口服剂和检测试剂用于预防、治疗和诊断动物疾病,是近年来抗体制备技术中新兴的研究领域。