鸡新城疫红细胞凝集抑制试验不同操作方法的比较

<正> 鸡新城疫康复鸡、隐性感染鸡和免疫后的鸡血清中都含有抗鸡新城疫病毒的抗体,有抑制鸡新城疫病毒凝集红细胞的特异作用,因此可以应用红细胞凝集抑制试验(简称HI)来诊断鸡是否患过鸡新城疫病和进行鸡群免疫状态的监测。目前HI试验有几种不同的做法,为了解这几种做法的HI效价是否存在明显差异,特对这几种方法进行比较。     

蛋鸡新城疫与低致病性禽流感发病特点与防控对策

<正>1非典型新城疫造成的原因新城疫病毒侵入机体咽喉部位定居和繁殖。根据机体与新城疫病毒之间力量对比,可表现为免疫、隐性感染、发病和死亡,隐性感染的鸡可不表现症状但排毒。鸡新城疫的临床表现决定于新城疫病毒的毒力和机体的免疫状态。当新城疫病毒强毒遭到免疫力降低的鸡群或新城疫病毒中毒遇到无免疫力的鸡群时表现非典型新城疫。     

七彩山鸡新城疫病毒的分离及生物学特性测定

用SPF鸡胚从疑似七彩山鸡新城疫病鸡群中分离到1株病毒，经HA和HI试验及病毒中和试验证明该分离毒为七彩山鸡新城疫病毒。动物回归试验表明，分离毒肌肉注射非免疫七彩山鸡能使之发病、死亡，出现与自然病例一致的症状与病变，但肌注SPF鸡只能感染，不出现临床症状。其生物学特性：NHAT为快、HOT为8min、ICPI为1．46、IVPI为1．6、MDT为76．8h，与鸡新城疫病毒相比，表现出不同的特性。     

新城疫分离株免疫保护试验

对经过鉴定的新城疫病毒穴NDV雪分离株,按国家兽医生物制品质量标准制备油乳剂灭活疫苗,在隔离器中接种SPF鸡,设标准LaSota疫苗株作对照,进行免疫保护实验。免疫后,每7d采血监测NDV抗体。免疫后3周,利用ND强毒分离毒和F48E9分别进行交叉攻毒实验,同时设SPF鸡对照。每天观察,及时剖检发病鸡,检察鸡群病变,以确定疫苗的保护性,以便对常规生产中的免疫失败进行原因分析。     

山东省鸡新城疫病毒弱毒株的初步分离

某鸡场饲养的罗曼产蛋鸡群爆发了典型的新城疫 ,采集病料接种11日龄SPF鸡胚进行病毒分离。经致鸡胚病变、回归试验、血凝和血凝抑制试验结果表明 ,所分离到的病毒系鸡新城疫病毒弱毒株 ,定名为NDV—S1。     

Immune protection assay of Newcastle disease live vaccine （LaSota Strain） against a genotype VII NDV isolate

从山东省发病鸡群分离鉴定了一株新城疫病毒（NDV）,命名为SDLY01。经蚀斑纯化后进行毒力测定和序列分析表明分离株SDLY01属于基因VII型NDV强毒。20只7日龄SPF鸡免疫新城疫活疫苗LaSota后14d分别用NDV标准强毒F48E8和分离株SDLY01攻毒,同时设同日龄SPF鸡为对照组,未免疫任何疫苗。攻毒后观察10d,免疫组在攻毒后食欲、精神均正常;对照组在攻毒后2～4d发病死亡,并表现ND典型的临床症状和病理变化。攻毒后第3、5、7、9d对免疫组试验鸡取喉头、泄殖腔棉拭进行病毒分离,F48E8攻毒组病毒分离均为NDV阴性,SDLY01攻毒组第5d病毒分离NDV阳性,第3、7和9d病毒分离阴性。本研究结果表明LaSota活疫苗对F48E8和SDLY01均能提供100％免疫保护,但不能完全抑制基因VIINDV分离株在体内的复制和排毒。     

ELISA检测鸡新城疫病毒特异性IgM抗体的研究

以新城疫病毒单克隆抗体包被板,用10％小牛血清-PBS封闭后,捕获尿囊液中的新城疫病毒作固相抗原.在此板上应用酶标抗鸡IgM单克隆抗体进行间接ELISA试验检侧鸡血清中新城疫病毒的特异性IgM抗体.试验证明该方法特异性强、敏惑性高.兔抗新城疫病毒阳性血清可特异性阻断反应,将新城疫病IgM阳性血清用2-ME处理可使ELISA反应呈阴性,与禽源多杀性巴氏杆菌鸡IgM阳性血清、鸡传染性支气管炎病毒IgM阳性血清无交叉反应.该试验可检测到La Sota免疫后3天鸡血清中的特异性IgM,对鸡新城疫病毒IgM阳性血清的检测效价可达1:320以上,并可检测到临床新城疫病鸡血清中的特异性IgM抗体.     

山东省鸡新城疫病毒弱毒株的初步分离

某鸡场饲养的罗曼产蛋鸡群爆发了典型的新城疫,采集病料接种１１日龄ＳＰＦ鸡胚进行病毒分离。经致鸡胚病变,回归试验、血凝和血凝抑制试验结果表明,所分离到的病毒系鸡新城疫病毒弱毒株,定名为ＮＤＶ－Ｓ１。     

分子筛凝胶浓缩新城疫病毒疫苗的研究

本文采用新型分子筛凝胶作为浓缩剂对新城疫病毒感染鸡胚的尿囊毒进行了浓缩实验,结果表明,该浓缩胶可使新城疫病毒浓缩９６．２４倍,病毒的损失量为８．６５％,用浓缩胶浓缩新城疫病毒后并制成灭活苗,浓缩苗进行鸡的免疫实验,以研究浓缩疫苗的免疫效果。浓缩新城疫抗原后制备的灭活苗在免疫后８天抗体水平迅速上升,ＨＩ为５．６,并持续上升,均高于对照组,说明浓缩疫苗可以使鸡产生较高的抗体水平。     

鸡新城疫诊断与防制的体会(下)

（续上期）如上所述，由于我们对鸡群采取了广泛和密集的疫苗接种，确实避免了急性型新城疫的爆发和损失，但却无法避免免疫鸡的带毒和排毒现象。事实上，很多表面上看来相当健康和平静的鸡群，其实都仍在带毒和排毒，如对这样的鸡群进行临诊上的检疫，很难有理由说它们不合格，但一旦进行精细的检疫，例如从泄殖腔、喉气管粘膜取样或将已包装好的禽肉反复冻融后取肉汁分离病毒，则不难分离到毒力不等的新城疫病毒（疫苗病毒或野外强毒）。由于带毒和排毒鸡群的普遍存在，无论对本场或其他鸡群都是一种危险的威胁，一旦鸡群内一些个体的免疫…     

Simulation of maternal immunity by inoculation of immune yolk preparations into the yolk sac of 1-day-old chickens.

Yolk harvested from eggs laid by hens hyperimmunized with killed Newcastle disease virus (NDV) was inoculated into the yolk sac of 1-day-old specific-pathogen-free (SPF) chickens. Serum hemagglutination-inhibition antibody titers reached maximum levels 1 to 4 days after yolk inoculation and declined at a rate similar to that reported for naturally acquired maternal antibody. Expected levels of immune interference were observed when yolk-inoculated chickens were vaccinated with a conventional oil-emulsion NDV vaccine. These results show that yolk-sac inoculation with yolk antibody is a suitable approach for producing maternally immune chickens for laboratory studies.     

Protection of chickens from Newcastle disease and infectious laryngotracheitis with a recombinant fowlpox virus co-expressing the F, HN genes of Newcastle disease virus and gB gene of infectious laryngotracheitis virus

A recombinant fowlpox virus (rFPV) coexpressing the Newcastle disease virus (NDV) fusion and hemagglutinin-neuraminidase genes and infectious laryngothracheitis virus (ILTV) glycoprotein B gene was constructed. This virus was then evaluated for its ability to protect specific-pathogen-free (SPF) chickens against clinical symptoms and death after challenge by virulent NDV and ILTV. SPF chickens were grouped and vaccinated with the rFPV and commercial NDV (La Sota) and ILTV attenuated live vaccine (Nobilis ILT), respectively. After challenge with NDV 10 days postvaccination, 70% of chickens vaccinated with rFPV were protected from death, whereas 100% of the commercial NDV-vaccinated chickens were protected from death. In contrast, 100% of the unvaccinated chickens died after challenge. After challenge with ILTV, both the rFPV and commercial ILTV-vaccinated chickens were completely protected from death and 70% of chickens were protected from respiratory signs. In comparison, 100% of the unvaccinated chickens developed severe respiratory disease and 10% of chickens died. The protective efficacy was also measured by the antibody responses and isolation of challenge viruses. Results showed that this rFPV could be a potential vaccine for preventing NDV and ILTV by a single immunization.     

法氏囊提取液对新城疫活疫苗免疫效力的影响

本文对鸡法氏囊提取液与ND活疫苗的使用方法进行了分析。通过血凝抑制抗体滴度的测定结果表明,预先接种鸡法氏囊提取液时,经肌肉注射、口服和点眼滴鼻接种方式均能提高ND疫苗免疫鸡的抗体产生能力,且经肌肉注射法最佳。但是,在整个试验期间,法氏囊提取液只能增强鸡群对ND活疫苗的早期免疫应答,而法氏囊提取液与ND活疫苗同时使用时更能刺激免疫鸡群的早期抗体产生水平。     

鸵鸟新城疫病毒野毒株的分离及其生物学特性

用鸡胚从疑似鸵鸟新城疫的送检病料中分离到 2株病毒。 2株病毒均能凝集鸡红细胞 ,且能被抗新城疫病毒 ( NDV)阳性血清抑制 ;用抗 NDV单抗 PEG夹心 ELISA测定 2株分离毒均为阳性。对其中 1株作进一步生物学特性鉴定 ,按照国际上规定的 NDV毒力判定标准测定的该毒株最低致死量致死鸡胚的平均死亡时间 ( MDT)为 54h,1日龄雏鸡脑内接种致病指数 ( ICPI)为 1 .88,6周龄雏鸡静脉接种致病指数 ( IVPI)为2 .75。结果表明 ,本分离株为鸵鸟新城疫病毒强毒     

不同新城疫弱毒苗对新城疫病毒强毒的免疫保护试验

用NDV的LaSota株、VG/GA 株、VH 株、PHY LMV 42株及Clone 30株弱毒疫苗免疫SPF鸡后 ,通过对雏鸡免疫后的血清抗体监测表明 ,4种弱毒疫苗激发雏鸡的抗新城疫病毒抗体滴度在 7log2水平以下 ,且差异不显著。通过以上 4种新城疫弱毒疫苗对新城疫病毒东台强毒株的免疫保护试验表明 :至少单独使用弱毒疫苗 ,不能对东台地区流行的新城疫病毒强毒株提供有效保护 ,保护率在 60 %～ 74%     

CAV与REV共感染SPF鸡对疫苗免疫反应的抑制作用

用1日龄SPF鸡人工感染鸡贫血病毒(CAV)和禽网状内皮增生病病毒(REV),探讨病毒感染对鸡体疫苗免疫反应的影响。结果表明,在用禽流感病毒(AIV,H5和H9)疫苗免疫后,CAV与REV单独感染均显著抑制了鸡体对H5和H9亚型禽流感病毒灭活疫苗的HI抗体反应,在CAV与REV共感染后,这种抑制作用更为明显。CAV单独感染后鸡体对新城疫病毒(NDV)和传染性法氏囊病病毒(IBDV)疫苗的免疫反应受到抑制,但与对照组在统计学上的差异不显著,然而,CAV可以显著加重REV感染对鸡体在NDV和IBDV疫苗免疫后抗体反应的抑制作用。从而证实CAV与REV共感染在疫苗免疫抑制上有协同作用。     

Protective antibody response produced by the chickens vaccinated with green coloured thermostable Newcastle disease virus

The efficacy of green-coloured (GC) I-2 Newcastle disease vaccine was determined in the present study. I-2 vaccine was mixed with a green coloured dye and stored at 4°C for 6 months while assayed for the virus infectivity at a monthly interval. Chickens were vaccinated with the GC vaccine by eye drop. Serum samples were collected from all birds before and after vaccination at weekly interval for 4 weeks and tested for haemagglutination-inhibition (HI) antibody against Newcastle disease virus (NDV). These chickens were challenged with NDV virulent strain four weeks after vaccination. The results showed that there was no difference between the infectivity titres of GC and uncoloured vaccines. However, chickens vaccinated with GC vaccine produced higher HI antibody titres than chickens vaccinated with uncoloured vaccine. Results from the challenge trial showed that all vaccinated chickens survived whereas all unvaccinated chickens died. The findings from this study have shown that the GC vaccine is safe and produced protective antibodies against NDV in vaccinated chickens. Wambura, P. N., 2008. Protective antibody response produced by the chickens vaccinated with green coloured thermostable Newcastle disease virus. Tropical Animal Health and Production.     

Oral vaccination of chickens against Newcastle disease with I-2 vaccine coated on oiled rice

Antibody response produced by Newcastle disease virus (NDV, strain I-2) when given orally through oiled rice to chickens was determined. Serum samples were collected before and at a weekly interval for 28 days after vaccination and tested for haemagglutination inhibition (HI) antibody to NDV. The results showed 7 days after vaccination HI antibody titre log₂ was 3.8. Moreover, 14 and 28 days after vaccination HI antibody titre log₂ reached 6.5 and 8.0, respectively. All unvaccinated chickens were negative to NDV antibody throughout the study. Significant finding from the present study is that 7 days after vaccination chickens had produced protective antibody against NDV; this is in contrast to previous studies. Therefore, I-2 vaccine coated on the oiled rice is efficacious as it protects chickens from challenge with NDV. Wambura, P. N., 2008. Oral vaccination of chickens against Newcastle disease with I-2 vaccine coated on oiled rice. Tropical Animal Health and Production.     

THE SEROLOGICAL RESPONSE OF CHICKENS TO AUSTRALIAN LENTOGENIC STRAINS OF NEWCASTLE DISEASE VIRUS

SUMMARY: Australian lentogenic Newcastle disease viruses were evaluated as uninactivated vaccines in Australian chickens, the response being evaluated by the production of haemagglutination-inhibition (HI) antibodies. Two viruses, V4 and PM9, induced high levels of antibody and were readily transmissible between chickens by contact exposure. Three other viruses were poorly immunogenic and poorly transmissible. Chickens vaccinated intramuscularly with the V4 strain produced higher HI antibody titres than chickens vaccinated by the orotracheal, intranasal and intraocular routes. HI antibody titres in chickens vaccinated with the V4 strain reached peak levels 3 to 5 weeks after vaccination and waned considerably during the next 2 to 4 weeks. However, low levels of HI antibody persisted for at least 36 weeks after vaccination. Intramuscular vaccination with the V4 strain of one-day-old chicks lacking maternal antibody to Newcastle disease virus resulted in 42–70% mortality and the survivors developed very high titres of HI antibody. Similar chickens inoculated orotracheally showed signs of depression and developed high titres of HI antibody, but there were no mortalities. Chickens 1-, 2-, 3- and 4-weeks-old and lacking maternally derived HI antibody to Newcastle disease virus suffered no adverse reaction to intramuscular or orotracheal vaccination. The antibody response of the 1-week-old chickens was considerably poorer than that of the older chickens. Following orotracheal vaccination with the V4 strain, chickens with low levels of maternally derived antibody responded with low levels of HI antibody. On the other hand, in the progeny of hens hyperimmunised with the V4 strain the production of active antibody following orotracheal vaccination was delayed until the level of passive antibody had declined considerably. There was no response to intramuscular vaccination in congenitally hyperimmune chickens. The minimum HI antibody inducing dose of V4 vaccine, when measured 3 weeks after vaccination of 6-weeks-old chickens, was 10^5.6 50% egg infectious doses.     

Reduced serologic response to Newcastle disease virus in broiler chickens exposed to a Chinese field strain of subgroup J avian leukosis virus

In this study, a Chinese field strain of subgroup J avian leukosis virus (ALV-J), NX0101, was studied for its immunosuppressive effects in both commercial broilers and SPF white Leghorn chickens infected at 1 day of age. Our data demonstrated that NX0101 induced much more significant body and immune organ weight loss in the infected commercial broiler chickens in an earlier age than that in the SPF white Leghorn chickens. At the same time antibody responses to vaccinations of Newcastle disease virus (NDV) and infectious bursa disease virus (IBDV) in the NX0101-infected chickens were also evaluated and compared between the commercial broiler chickens and the SPF white Leghorn chickens. Compared with the control group of chickens, the hemagglutination inhibition (HI) antibody response to NDV vaccines was significantly reduced in the NX0101-infected commercial broiler chickens from as early as 20 days after vaccination. However, no significant difference in HI antibody response was seen when HI titers reached their peaks in the NX0101-inoculated and control SPF white Leghorn chickens, except it declined significantly faster in infected birds. Neither of these two types of chickens showed significant decrease of antibody response to IBDV vaccination. Herein, we conclude that this NX0101 strain of ALV-J could selectively suppress humoral immune reactions to NDV, especially in broilers. But challenge experiments were not conducted and, therefore, it cannot be known if decreased antibody levels correlated with decreased protection against NDV in this case.