Isolation and biological properties of virulent subpopulations from lentogenic Newcastle disease virus strains

From each of two lentogenic Newcastle disease virus (NDV) strains of the type LaSota and Hitchner B1 a virulent subpopulation could be obtained. The two subpopulations were—in comparison to the two parent viruses—more resistant to the lipid solvent chloroform and more stable against thermal degradation. Also, the glycoproteins haemagglutinin and F (fusion) were more stable against thermal inactivation. Electron microscopic observations revealed in terms of size and morphology all of the characteristics of NDV. Both subpopulations possessed, however, the same elution kinetics as their respective parent strains. The intracerebral and intravenous pathogenicity indices as well as the mean death times of the two subpopulations allow to classify these viruses as virulent Newcastle disease viruses.     

3株新城疫病毒毒力的测定

新城疫是由新城疫病毒引起鸡的一种高度接触性传染病。为了探讨本地有关区域 NDV毒力分布情况 ,将自 1 997年以来 ,从珠江三角洲不同地区分离获得的 3株 NDV的鸡胚平均致死时间 ( MDT)、1日龄雏鸡脑内致病指数 ( ICPI)和 6周龄鸡静脉内致病指数 ( IVPI)进行测定。1　材料与方法1 .1　病毒　ND野毒株 ,共 3株 ,包括 N株 (分离自南边 )、S株 (分离自狮山 )、L株 (分离自罗定 )均由本实验室分离、鉴定保存。1 .2　实验动物　 1 0日龄鸡胚、1日龄易感雏鸡 ,6周龄非 ND免疫雏鸡均由佛山燎原鸡场提供。1 .3　毒株的鸡胚传代　将 3株 ND…     

Genetic relationships among lentogenic strains of Newcastle disease virus

Purified RNA from five lentogenic Newcastle disease virus (NDV) strains (B1, England-F, Nebraska, Queensland V4, and Ulster) was subjected to oligonucleotide fingerprinting analysis. This technique could resolve over 100 oligonucleotides per virus strain, of which 50 to 80 were unique. Although the fingerprints generated from these viral RNAs exhibited similar distribution patterns of non-unique RNA fragments, all could be distinguished from one another on the basis of the migration of unique RNA fragments. It is concluded that oligonucleotide mapping is a useful method of unambiguously identifying individual lentogenic strains of NDV and that this technique may serve as a means of characterizing highly pathogenic NDV isolates.     

新城疫病毒在鸡胚中增殖特性的研究

本研究将接种到9～10日龄鸡胚中的鸡新城疫病毒(NDV)La Sota或V4株弱毒培养24h,病毒滴度可增加106以上,培养108 h,病毒滴度则增加至10 10倍以上.NDV在有NDV母源抗体的普通鸡胚与无抗体的SPF鸡胚中的增殖无显著差异;盲传对于检验NDV没有意义.La Sota或V4株病毒悬液在理论上稀释至每胚接种量只有数个或1个病毒时,仍可引起鸡胚感染.低毒量条件下固定病毒的数量,随着接种胚数的增加,感染率随之下降,但感染数在一定范围内波动,没有减少趋势.以上结果证实数个甚至1个活的NDV粒子即可引起鸡胚感染,这对现地检测NDV具有重要意义.     

鸭源新城疫病毒的分离与鉴定

2005年5月河北某地区25日龄金定鸭突然发生一起以腹泻、病死率较高、腺胃黏膜出血或有纤维素性渗出物、肠道黏膜出血及胰腺偶见白色坏死点为特征的新的传染病,给养鸭业造成了一定的经济损失.通过病毒分离、电镜形态观察、血清学鉴定、理化特性和致病性试验等,对该病原进行了分离和特性鉴定,为防控该病提供理论基础.     

Vaccination with CDF-66 strain of newcastle disease virus

Newcastle disease vaccine CDF-66 has been proved effective by the drinking water route. A chicken dose of 10^6.2 EID₅₀ in 12 ml of water containing 2.5% skimmed milk was found to be effective. This produced sufficient GMHI antibody response (105) and 86.6% of birds withstood challenge with the Mukteswar strain of virulent virus. The in-contact birds did not show either immune or antibody response. The vaccinated birds excreted vaccine virus only from the respiratory tract not from cloaca. The vaccine given in drinking water protected only 50% of chickens when it was given 168 h prior to challenge virus. With lesser intervals the protection was negligible. Diluted vaccine when held for 2 h at 37°C and room temperature did not show a drop in potency.

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Vacunacion Con La Cepa Viral CDF 66 De Newcastle

Resumen La vacuna CDF 66 contra newcastle suministrada en el agua de bebida ha tenido buen resultado. Una dosis individual de 10^6.2 DIE₅₀ en 12 ml. de agua estabilizada con 2.5% de leche descremada es suficiente para conferir una buena inmunidad. Esta produjo suficientes anticuerpos GMHI (105), y 86.6% de aves resistieron la descarga de virus virulento cepa Mukteswar. Las aves en contacto no mostraron inmunidad ni respuesta de anticuerpos. Las aves vacunadas excretaron virus del tracto respiratorio únicamente. La vacuna en el agua de bebida protegió el 50% de aves cuando se suministró 168 h. antes de la descarga con virus virulento. Con intervalos menores el efecto fue casi nulo. La vacuna diluida no perdió potencia a una temperatura de 37°C por 2 h.

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Vaccination Avec La Souche Virale CDF-66 De La Maladie De Newcastle

Résumé Le vaccin contre la maladie de Newcastle à l'aide de la souche CDF-66 dans l'eau de boisson a donné de bons résultats. La dose par animal de 10^6.2 EID₅₀ dans 12 ml d'eau contenant 2,5 p. 100 de lait écrèmé s'est révélé efficace. Elle a produit une réponse en anticorps GMHI (105) suffisante et 86,6 p. 100 des oiseaux ont résisté à l'épreuve consistant en l'inoculation d'un virus vivant non atténué de la souche Mukteswar. Les oiseaux non traités n'ont montré ni immunité ni réponse antigénique. Les oiseaux vaccinés ont excrété le virus-vaccin par les voies respiratoires seulement et non par le cloaque. Le vaccin administré dans l'eau de boisson a protégé seulement 50 p. 100 des poulets lorsqu'il a été administré 168 heures avant l'epreuve virale. A intervalles moindres, la protection a été négligebale. La dilution vaccinale, lorsque conservée pendant 2 heures à 37°C ou à la température du local, n'a rien perdu de son efficacité.

     

抗新城疫病毒血凝素单克隆抗体的研制

新城疫是由新城疫病毒(NDV ) 引起的一种主要侵害鸡和火鸡的急性、高度接触性传染病, 危害极其严重.根据流行特点、临床表现和病理变化可对典型性新城疫作出诊断,但对非典型性新城疫的诊断及该病的确诊需借助于病毒的分离鉴定、血清学方法、直接病毒抗原检测等实验室手段.研制抗NDV单抗将为NDV的检测发挥重要作用,曹殿军等[1]已制备出抗新城疫病毒糖蛋白单克隆抗体,但抗NDV血凝素单抗国内少有报道,鉴此,本实验室用PEG融合技术,研制出分泌抗NDV血凝素蛋白杂交瘤细胞株.     

马立克氏病病毒感染鸡对新城疫疫苗和法氏囊疫苗免疫反应的影响

研究了马立克氏病病毒(MDV)不同致病型毒株感染鸡后对新城疫病毒(NDV)和传染性法氏囊病病毒(IBDV)疫苗抗体反应的影响。结果表明，2周龄接种MDV超强毒株RBIB和7日龄接种MDV强毒株GA的鸡HI效价均显著低于对照鸡，但在本试验条件下尚未发现MDV感染对法氏囊病病毒疫苗免疫效果有显著影响。     

新城疫病毒弱毒株对鹅的免疫效力试验

用新城疫病毒(NDV)疫苗株La Sota和NDV基因Ⅶ型致弱株AND/Ⅶ为免疫原,分别以活苗和灭活苗的形式免疫2周龄试验鹅,免疫后第7、14天和21天采血分离血清用于抗体效价测定.免疫后3周以基因Ⅶ型强毒分离株JS2-06进行攻毒,攻毒后每天观察各组鹅的发病情况,并于第2、4天和7天分别采集喉气管和泄殖腔棉拭样品,用于病毒的分离.试验结果表明,活疫苗免疫后不能产生有效的免疫应答,而灭活苗免疫鹅对强毒的感染具有较强的抵抗能力,但基因Ⅶ型致弱株AND/Ⅶ能更有效抑制鹅体的排毒,其免疫效力明显高于疫苗株La Sota.     

Cytopathic effect of lentogenic strains of Newcastle disease virus in cultured chicken muscle cells

The ability of lentogenic strains of Newcastle disease virus (NDV) to inhibit the development of embryonic chicken muscle cells in vitro is described. In contrast to the uninfected control, which displayed a thick network of branched multinucleated muscle fibers, cultures infected with NDV exhibited a severely reduced potential to differentiate. The few differentiated muscle fibers that formed in the presence of NDV were poorly developed, displayed an attenuated morphology, and detached from the substrate 2 to 3 days before uninfected cultures showed signs of cell aging. When NDV-infected cultures were stained before muscle fiber detachment with a fluorescent-labeled NDV-specific antibody, the infected muscle fibers reacted strongly relative to neighboring myoblasts and fibroblasts. All six strains of NDV induced a similar inhibitory effect, and no strain-specific differences were detectable. These results indicate that differentiating muscle fibers are highly susceptible to infection by NDV in vitro and that this tissue exhibits an early NDV-induced cytopathic effect.     

新城疫病毒HN蛋白抗原表位分析及结构域基因原核表达

以本实验室构建的含新城疫病毒血凝素-神经氨酸酶基因重组质粒为模板，设计引物通过PCR和重叠-延伸PCR扩增，获得HNa、HNb和HNa-L-b三种抗原结构域基因片段，BamHⅠ／HindⅢ双酶切定向克隆到原核表达载体pET32c，获得重组质粒分别命名为pET32c—HNa、pET32c-HNb和pET32c．HNa—L—b。重组质粒转化大肠杆菌感受态细胞BL21，筛选出阳性克隆，诱导表达并取产物进行分析。结果表明，HNa、HNb、HNa-L-b结构域基因片段均获得了融合表达。Western-blotting分析证实表达产物HNa和HNb与NDV阳性血清具有免疫反应性。本试验结果为进一步研究HN蛋白抗原结构域的免疫原性以及HN蛋白与F蛋白在细胞融合中的相互作用奠定了基础。     

Response of domestic geese to lentogenic and velogenic strains of Newcastle disease virus

A total of 54 domestic white meat-type geese were included in vaccination/challenge trials to evaluate susceptibility to disease and humoral immune responses using the haemagglutination inhibition (HI) and virus neutralization (VN) tests against Newcastle disease (ND). Two groups of twenty geese, five weeks of age, were conjunctivally vaccinated with either 100 x 10(6) or 2.5 x 10(6) EID50 (egg infectious dose 50 per cent) per bird of live La Sota virus, respectively, and 14 geese remained unvaccinated. At 15 weeks of age all vaccinated geese and seven unvaccinated geese were subcutaneously injected with 0.5 ml of inactivated oil emulsion ND vaccine, whereas seven geese remained as negative controls. At an age of 20 weeks, all 54 geese were challenged with 10(8.0) EID50 per bird of the viscerotropic velogenic NDV strain Herts 33/56. Live virus application as well as the oil emulsion vaccine did not induce discernible clinical signs and have no detrimental effect on body weight gains. At days 1, 3, 5, 8, 13, 16, 20, 23 and 27 after the application of lentogenic vaccine pharyngeal and cloacal swabs were taken, after challenge samples were taken at days 2, 5 and 8. Lentogenic as well as velogenic virus were never reisolated. Low and shortlived antibody responses post vaccination were equally well measured in HI and VN tests. Only two out of seven unvaccinated but challenged geese developed signs of ND whereas all vaccinated/challenged geese remained normal but developed high to moderate levels of HI and VN antibodies. Since domestic geese do not readily excrete NDV's in detectable amounts and since they do not contain detectable amounts of the challenge virus fourteen days post challenge in their tissues the assumption is promoted that geese do not play a major role in the epidemiology of Newcastle disease.     

我国部分地区新城疫病毒的流行现状分析

从近年来由我国不同地区、不同宿主分离到的新城疫病毒(NDV)野外分离株中,选取其中具有代表性的16株,用RT PCR技术扩增其F基因重要的功能区片段,进一步进行克隆和序列测定.参照国内外已发表的部分毒株的F基因序列,构建了59株NDV的遗传进化树,分析其毒株间的遗传进化关系.序列分析表明,所扩增的目的片段长度为535bp,所分离的毒株在分裂位点的氨基酸顺序为112R-R-Q/R-K/R-R-F117,均相当于NDV的强毒株.通过遗传进化树分析表明,16株分离株中有13株为基因Ⅶ型NDV,说明目前基因Ⅶ型NDV所引起的新城疫在国内呈流行趋势.     

Pathological changes of tracheal mucosa in chickens infected with lentogenic Newcastle disease virus

Forty-day-old specific-pathogen-free chickens were exposed by aerosol to lentogenic Newcastle disease virus (NDV) and observed for 24 days for pathological changes in the tracheal mucociliary system. Specific fluorescence of NDV antigen was observed through day 5 postexposure (PE) in the cytoplasm of the tracheal epithelium and desquamated epithelium in the lumen. On day 1 PE, scanning electron microscopy revealed hypertrophy of goblet cells and small patches of the deciliated epithelium scattered mainly around the openings of mucous glands. The deciliated area of tracheal surface increased through day 4 PE. Light microscopy showed small vacuoles containing lymphocytes and heterophils in the epithelial layer. Immature epithelium proliferated in some areas. On days 5 and 6 PE, ciliated areas of the trachea tended to increase as a result of regeneration of the epithelium, still leaving many nonciliated patches of various sizes. On and after day 8 PE, there remained plaques with nonciliated flat epithelium, but most areas were covered with well-ciliated epithelium. Non-ciliated plaques were observed until day 24 PE, but they gradually decreased in size. These plaques were covered by a single layer of flat epithelium and were formed upon lymph follicles in subepithelial tissue.