鸽禽I型副粘病毒油佐剂灭活苗对肉种鸽免疫效果观察

用鸽Ａ／ＰＭＶ－１ＳＸ株（简称ＳＸ株）制成的油佐剂灭活苗与ＮＤＶ油佐剂灭活苗分别免疫肉种鸽,免疫后２０天抗体水平达到峰值,抗体水平在３ｌｏｇ２以上维持３０天以上。免疫后４０天用鸽ＳＸ株和新城疫强毒对两种疫苗免疫鸽分别进行攻击,两种疫苗免疫组对ＮＤＶ强毒攻击的保护率均为１００％,鸽Ａ／ＰＭＶ－１油佐剂灭活苗免疫组对鸽ＳＸ株攻击的保护率为１００％,而ＮＤＶ油佐剂灭活苗免疫组对鸽ＳＸ株攻击的保护率仅为６６７％。鸽Ａ／ＰＭＶ－１油佐剂灭活苗田间试验抽检４８份肉种鸽血清,抗体平均值为４３０±１１６ｌｏｇ２。     

鸽禽Ⅰ型副粘病毒油佐剂灭活苗对肉种鸽免疫效果观察

用鸽Ａ／ＰＭＶ－１ ＳＸ株制成的油佐剂灭活苗与ＮＤＶ油佐剂灭活苗分别免疫肉种鸽,免疫后２０天抗体水平达到峰值,抗体水平在３ｌｏｇ２以上维持３０天以上。免疫后４０天鸽ＳＸ株和新城疫强毒对两种疫苗免疫鸽分别进行攻击,两种疫苗免疫组对ＮＤＶ强毒攻击的保护率均为１００％,鸽Ａ／ＰＭＶ－１油佐剂灭活苗免疫组对鸽ＳＸ株攻击的保护率为１００％,而ＮＤＶ油佐剂灭活苗免疫组对鸽ＳＸ株攻击的保护率仅为６６．７％。     

鸡新城疫—减蛋综合征蜂胶佐剂灭活二联苗的研究

首次用蜂胶作为佐剂，研制出鸡新城疫－减蛋综合征（ＮＤ－ＥＤＳ）二联灭活 ，在开产前１个月左右，用该苗按０．５ｍｌ／只的剂量皮下或肌肉注射免疫蛋鸡，接种后２０ｄ测ＮＤ抗体（ＨＩ法）效价为６．２５－７．０（ｌｏｇ２，）ＥＤＳ－７６抗体（ＨＩ法）效价为６．５－８．０，用ＮＤ－ＥＤＳ油乳剂灭活二联苗做对照，进行免疫持续期平行测定对比试验，免疫后１８０ｄ，蜂胶苗免疫组鸡的ＮＤ抗体滴度为８．５－８．７，ＥＤＳ     

新城疫与传染性法氏囊疫苗疫苗对雏鸡联合免疫的研究

将１１４只１１日龄海赛克斯褐公鸡随机分成５组：Ⅰ组为空白对照，Ⅱ组用新城疫弱毒苗和ＮＤ油佐剂灭活苗同时免疫，Ⅱ组用传染性法氏囊病地方毒株油佐剂灭活苗免疫，Ⅳ组用ＮＤ弱毒苗，ＮＤ油佐剂灭活苗，ＩＢＤ地方毒株油佐剂灭活苗同时免疫，Ｖ组在Ⅳ组的基础上增加ＩＢＤ弱毒苗免疫。５组鸡分别隔离饲养，定期测定血清ＮＤＨＩ效价与ＩＢＤ琼扩效价。选择时机用强毒攻击，观察各组鸡的保护情况。结果：（１）Ⅱ、Ⅳ、Ⅴ组鸡均能     

鸡新城疫／传染性支气管炎二联油乳剂灭活疫苗试验研究报告（Ⅲ）

用同胚培养的含ＮＤＶ、ＩＢＶ胚液制备的二联油乳剂灭活苗，经免疫效力试验表明，用５０倍稀释的鸡二联灭活苗，０．５ｍｌ／只剂量免疫１８日龄鸡，４周后试验免疫鸡分别进行ＮＤＶ、ＩＢＶ强毒攻击，攻毒保护率在５０％以上，证明０．５ｍｌ剂量含有５０个半数保护量（５０个ＰＤ５０），ＮＤＨＩ抗体≥１：１６，ＩＢＶＮ抗体≥１：８便可保护。三批疫苗免疫１８日龄鸡，０．５ｍｌ／只组免疫鸡体现人有效抗体可维持１６周以上，     

新城疫与传染性法氏囊病疫苗对雏鸡联合免疫的研究

将１１４只１１日龄海赛克斯褐公鸡随机分成５组：Ⅰ组为空白对照，Ⅱ组用新城疫（ＮＤ）弱毒苗和ＮＤ油佐剂灭活苗同时免疫，Ⅲ组用传染性法氏囊病（ＩＢＤ）地方毒株油佐剂灭活苗免疫，Ⅳ组用ＮＤ弱毒苗、ＮＤ油佐剂灭活苗、ＩＢＤ地方毒株油佐剂灭活苗同时免疫，Ⅴ组在Ⅳ组的基础上增加ＩＢＤ弱毒苗免疫。５组鸡分别隔离饲养，定期测定血清ＮＤＨＩ效价与ＩＢＤ琼扩效价。选择时机用强毒攻击，观察各组鸡的保护情况。结果：（１）Ⅱ、Ⅳ、Ⅴ组鸡均能很好地抵抗ＮＤＶ强毒的攻击，Ⅲ、Ⅳ、Ⅴ组鸡无论对ＩＢＤＶ人工感染，还是自然感染，均有坚强的保护力；（２）ＩＢＤ地方毒株油佐剂灭活疫苗不干扰ＮＤ疫苗的免疫应答；（３）ＩＢＤ弱毒苗对ＮＤ疫苗的免疫应答有一定影响，在免疫初期能促进机体尽快对ＮＤ疫苗产生应答，但后期ＮＤ疫苗的ＨＩ抗体降低较快；（４）ＮＤ疫苗对ＩＢＤ疫苗的免疫效果无显著影响。作者提出，对雏鸡首免（无论有、无母源抗体）可采用ＮＤ弱毒苗＋ＮＤ油佐剂灭活苗＋ＩＢＤ（地方毒株）油佐剂灭活苗同时进行免疫，至少可以保护到上笼后或开产前免受ＮＤＶ和ＩＢＤＶ的感染，既简化了免疫程序，又可减少对鸡群的应激，值得推广应用。     

鸡新城疫,减蛋综合征,传染性支气管炎三联油佐剂灭活苗的研究…

用新城疫病毒克隆７９株，减蛋综合征病毒ＮＥ４株和传染支气管炎病毒Ｍ１株分别接种于鸡胚和鸭胚，并收取其鸡，鸭胚尿囊液毒，经甲醛灭活，按一定经例配比，以矿物油为佐剂制成三联油佐剂灭活苗。本苗接种于产蛋后备鸡，免疫后７天产生免疫应答，免疫后３０天保护率达９０％－１００％，免疫后六个月攻毒，ＮＤ和ＩＢ保护率１００％，ＥＤＳ７６为９５％。１９９２－１９９４上半年，本苗在江苏，安徽，山东等省的一些鸡场免疫十万     

鸡用天然缓释免疫增强剂的研究：天然缓释免疫增强剂对IBD等4？…

对７日龄艾维茵雏鸡投服天然缓释免疫增强剂１丸,１０日龄时分别用ＩＢＤ、ＮＤ、ＩＢ弱毒疫苗及ＮＤ油乳剂灭活苗免疫;免疫后７、１５、２５、３５一５５ｄ定期检测抗体效价;弱毒苗免疫后３５ｄ、灭活苗免疫后５５ｄ分别用强毒攻击。结果各试验组抗体效价显著高于对照组,试验组攻毒保护率提高１６．７￣３３．３个百分点。表明该缓释免疫增强剂能显著增强雏鸡的免疫应答,提高疫苗的免疫效力。     

用吐温80制备新城疫病毒乳化灭活疫苗

用１０％（Ｗ／Ｖ）吐温８０和０．０５％（Ｖ／Ｖ）福尔马林制备新城疫病毒（ＮＤＶ）水－油－水乳化（ＷＯＷＥ）灭活疫苗，其抗原亚单位平均分子量为３０７ＫＤ，油与水的比例为１：２，乳化的ＮＤＶ可单独应用，也构成ＷＯＷＥ四联疫苗。用４４．７μｌ去污剂处理的ＮＤＶ尿囊液（ＮＤＶ－ＡＦ）为一个剂量的单价苗，用其免疫５周龄商品鸡后７周攻毒，可获完全保护。用１７８．６和８９．３μｌ去污处理的ＮＤＶ－ＡＦ苗接种４周     

MD二价油佐剂灭活疫苗免疫的研究

自ＭＤ病鸡分离到的强致病力毒株，经鉴定为Ⅰ型ＭＤＶ，与Ⅲ型ＨＶＴ（ＡＨＶＴ１７）组合制成二价油佐剂灭活苗，经各项筛选试验，证明二价灭苗的免疫效果比任何一种单价苗好。免疫剂量在各１５００－３０００ＰＦＵ时，免疫保护率多数在４０％以下，免疫剂量在各１５００－３０００ＰＦＵ时，免疫保护率多数在４０％以下，且时高时低，而将ＰＦＵ提高到各５０００时，其免疫保护率即可提高到７２－８１．１７％。ＨＶＴ单价苗的免     

Study on Synergistic Effect of VA5 Immunopotentiator on Pigeon NDV Inactivated Vaccine

This study was designed to evaluate the synergistic effect of VA5 immunopotentiator on pigeon ND4416 inactivated vaccine.160 healthy pigeons were randomly divided into four groups,including ND4416 strain inactivated vaccine group (NDV group),ND4416 strain inactivated vaccine and immunopotentiator (VA5) mixed group (NDV+VA5 group),La Sota inactivated vaccine group (La Sota group) and normal saline as the control group (C group),to assess their vaccine efficacy against virulent pigeon NDV by serological analysis and animal testing.Pigeons sera were collected at different time points after immunization and measured the HI antibody titer of each group.The results showed that VA5 immunopotentiator significantly improved the serum antibody level of pigeons immunized with pigeon Newcastle disease inactivated vaccine (P<0.05).In addition,comparative test of spleen lymphocyte transformation was conducted at various time points after immunization.The results indicated that VA5 effectively stimulated the lymphocyte transformation of immune pigeon.Pigeons in each groups were challenged with ND4416 strain at the 30th,90th and 180th d after immunization.The results presented that the NDV+VA5 group had 100% protection rate and higher than La Sota group.The duration of immunization test showed that the antibody titer of NDV+VA5 group reached peak 11.20log₂ at the 21st d,remained 7.50log₂ at 180th d,and the protection rate remained 100% at 180th d.It indicated that VA5 immunopotentiator sustained the immune duration of pigeon NDV vaccine up to 180 d.Moreover,the in vitro detoxification test results suggested that VA5 immunopotentiator reduced the in vitro detoxification cycle of pigeons after challenge.Overall,this study suggested VA5 immunopotentiator could significantly improve the immune efficacy of pigeon Newcastle disease inactivated vaccine,which provided a basis for further enhancing the efficacy of Newcastle disease vaccine,as well increased experimental data for the application of immunopotentiators.     

VA5免疫增强剂对鸽新城疫病毒灭活疫苗的增效作用研究

试验旨在探究VA5免疫增强剂对鸽新城疫病毒（NDV）灭活疫苗（ND4416株）的免疫增效作用。选取160只健康鸽随机分为4组：ND4416株灭活疫苗组（NDV组）、ND4416株灭活疫苗与免疫增强剂（VA5）混合组（NDV+VA5组）、La Sota灭活疫苗组（La Sota组）及生理盐水空白对照组（C组）,进行免疫效力及免疫持续期试验。采用血凝抑制试验检测各组鸽免疫后不同时间点的血清抗体效价,结果显示,VA5免疫增强剂能够显著提高鸽NDV灭活疫苗血清抗体水平（P<0.05）。脾脏淋巴细胞转化试验结果显示,VA5能够有效刺激免疫鸽的淋巴细胞转化。在免疫后第30、90、180天,各组鸽子采用ND4416株进行攻毒试验,结果显示,NDV+VA5组在3个时间点的攻毒保护率均为100%,均高于La Sota组。免疫持续期试验结果显示,NDV+VA5组血清抗体水平在免疫后第21天达峰值11.20log₂,第180天仍能达到7.50log₂,攻毒保护率达100%,表明VA5免疫增强剂可延长鸽新城疫疫苗的免疫效力持续期至180 d。攻毒后体外排毒检测结果表明,VA5免疫增强剂能够缩短攻毒后鸽的体外排毒周期,减少体外排毒。综上,VA5免疫增强剂能够显著提高鸽NDV灭活疫苗的免疫效果,为研制效果更好的鸽NDV疫苗提供了基础依据,同时也为免疫增强剂的应用增加了试验数据。     

Effect of the Immunity Efficacy of IBD Immune Complex Vaccine on ND La Sota Vaccine

1-day-old SPF chickens and commercial Jingfen chickens were vaccinated with IBD immune complex(IC) vaccine, NDV La Sota vaccine were inoculated simultaneously every one week and every two weeks.NDV La Sota immunization alone was as the control group.At the 2nd, 3rd, 4th and 5th week post inoculation, blood samples were taken and the ND HI antibody were tested.Experimental chickens were challenged with high virulent NDV at the 5th week post inoculation, the protective rate of each group was calculated.The results showed the ND HI antibody were not significant different in the combined immunization of IBD IC vaccine priming and NDV La Sota vaccine boost and NDV La Sota vaccine alone immunization (P>0.05).The results indicated that IBD IC vaccine has no immunosuppression on NDV La Sota vaccine in SPF chickens and commercial Jingfen chickens.     

基于新城疫病毒V蛋白鉴别鸡群感染与免疫的间接ELISA方法的建立

本研究以新城疫病毒(NDV)V蛋白羧基端结构域(Vc)的重组蛋白为包被抗原,建立了用于检测NDV V蛋白抗体的间接ELISA方法,并采用该方法检测了鸡群免疫或接毒后血清中的V蛋白抗体水平。结果显示:两组不同NDV灭活疫苗组在免疫后的3周内检测结果均为阴性;两组灭活疫苗免疫3周后再人工感染NDV强毒的鸡群,攻毒后第7、14和21 d,NDV阳性率分别为60%、80%、70%和50%、80%、70%;两组不同的NDV弱毒疫苗免疫组鸡群,仅在免疫后第21 d阳性率分别为20%和10%。以上结果表明,NDV疫苗免疫组与强毒感染组的V蛋白抗体阳性率存在明显差异,本方法可在群体水平上区分新城疫疫苗免疫与强毒感染鸡群,为NDV血清学诊断和流行病学调查提供了一种新的检测手段。     

Evaluation of efficacy of Vero cell-adapted,thermostable Newcastle disease vaccine in broilers

The main theme of this project was to develop a Vero cell-adapted, thermostable NDV I-2 vaccine and evaluate its efficacy against challenge infection. For this purpose, serial passages of virus were done in Vero cell line up to 13 times and after each passage samples were subjected to heat treatment at 56°C for 40 min. After 13 passages, the virus was completely adapted on Vero cell line and cytopathic effects were observed, including syncytial formation, rounding, degeneration, and detachment of cells. Hemagglutination and infectivity titers showed that the virus was thermostable after each passages in Vero cell line. One-day-old broiler chicks (Group 1) were vaccinated orally with thermostable NDV I-2 vaccine. A commercially available thermolabile NDV LaSota was used in Group 2 used as positive control. NDV I-2 vaccine produced maximum % inhibition migration at d 6 (50%) as compared with LaSota ND vaccine (i.e., 32%). On encounter with virulent NDV I-2, 100% safety was accomplished in group 1 and 60% in case of group 2. All the birds in the control negative group had died. This study led to the conclusion that thermostable Vero cell adapted I-2 strain vaccine resulted in better immunization in broiler birds than obtained by the use of commercially available thermolabile vaccines.     

Immunosuppression and histopathological changes in the bursa of fabricius associated with infectious bursal disease vaccination in chicken

The effect of the infectious bursal disease (IBD) live virus vaccine on the immune response of chicken was evaluated by the assessment of antibody response following vaccination as well as resistance to challenge with virulent virus. Birds were vaccinated at various ages and later challenged with a heterologous vaccine (NDV) or wild-type IBD virus. The BF was examined for histological changes at regular intervals. Antibody levels to NDV were monitored.

Significantly higher mortality rates were observed in birds vaccinated with IBD vaccine than unvaccinated birds (P < 0.01) following challenge, BF from vaccinated birds showed marked lymphocyte depletion and cellular infiltration with mononuclear cells.

Intraocular NDV (NDV-i/o) vaccine given at day old largely prevented the immunodepressive effect of IBD vaccination on NDV vaccine. Groups that received IBD vaccine on day 14 but no NDV i/o suffered higher mortality (41.2%) and showed lower antibody response than those vaccinated on day 1 (0%) or controls which did not receive IBDV (11.8%).     

禽Ⅰ型副粘病毒各种禽源分离株的免疫原性

分别制备了源于鸡、鹅、鸽、鹌鹑、孔雀、画眉鸟、珍珠鸡的禽型副粘病毒(APMV-1)7个强毒分离毒株和商品弱毒疫苗株克隆30(C30)的灭活油乳剂苗,并用这8种灭活油乳剂苗和C30株的活疫苗分别在鸡、鹌鹑、鹅和鸽进行了免疫及交叉攻毒保护试验。免疫后(PI)分别测定试验禽血清的新城疫病毒(NDV)血凝抑制(HI)抗体的滴度,并于PI 5周用强毒株进行攻毒。结果表明,鸽的HI抗体几何平均滴度(MAT)为9.00~10.0 log2,鸡的为7.13~7.63 log2,鹌鹑的为5.00~5.13 log2,鹅对灭活油乳剂苗的为5.63~6.38 log2、而对C30株活疫苗的仅为3.38log2;除了C30株活疫苗免疫鹅提供的保护率比较低(20%)外,8种油乳剂苗都能对同源或者异源强毒株的攻毒提供比较高的保护率(66.75%~100%)。研究结果表明,经典疫苗株C30与近年来从各种不同禽类分离的致病性APMV-1野毒株之间、不同禽源分离株之间的抗原性,以及各种不同禽源分离株的之间的免疫原性差异均不大。     

鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活苗(La Sota株+M41株+Re-9株)应用效果研究

鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活苗(La Sota株+M41株+Re-9株)(三联苗(Re-9株))为国内首个采用基因重组H9亚型禽流感疫苗株研制的新支流三联灭活疫苗。为研究疫苗上市后的实际应用效果,使用7日龄AA肉鸡和260日龄产蛋期蛋鸡评价三联苗(Re-9株)有效性。结果显示:肉鸡免疫后14 d即可激发抗体,免疫后21 d,新城疫病毒(NDV)、传染性支气管炎病毒(IBV)、H9亚型禽流感病毒(H9 AIV)的血凝抑制(HI)抗体分别达8.6log2、6.2log2、8.3log2,出栏时仍维持在较高水平;于免疫后14 d抽样进行NDV、H9 AIV攻毒,对照组均100%发病或排毒,免疫鸡均可获得100%保护;在蛋鸡上,免疫三联苗(Re-9株)后28 d,NDV、IBV、H9 AIV的HI抗体效价分别达12.0log2、7.8log2、12.2log2,免疫后6个月仍可以维持在较高水平。结果表明:三联苗(Re-9株)在实际应用中免疫效果较好,本研究将为鸡新城疫、传染性支气管炎、H9亚型禽流感的防控提供有力支撑。     

麻鸡副黏病毒ZH-1株F基因核酸疫苗的构建及其免疫效果

应用RT-PCR方法从麻鸡副黏病毒ZH-1株中扩增F基因并克隆入pGEM○R-T载体,经序列测定、分析,结果显示ZH-1株F基因与NDV国家标准强毒F48E9株的核苷酸、氨基酸序列同源性均达到99%。随后将F基因克隆入真核表达载体pCI-neo,构建真核表达质粒pCI-F,pCI-F体外转染Vero细胞,能在Vero细胞中表达,利用pCI-F质粒进行动物试验,结果表明雏鸡免疫14 d后在体内可检测到特异性抗体,pCI-F质粒二次免疫雏鸡后,对NDV强毒的攻毒保护率为73%,这一结果提示利用F基因构建的核酸疫苗具有重要的开发前景。     

The resistance of meat chickens vaccinated by aerosol with a live V4 Newcastle disease virus vaccine in the field to challenge with a velogenic Newcastle disease virus

Meat chickens housed on a commercial broiler farm in Australia were vaccinated once at 10 to 11 days-of-age by aerosol with live V4 Newcastle disease virus (NDV) vaccine. Groups of vaccinated and unvaccinated birds were flown to Malaysia, where they were challenged with a virulent strain of NDV. Survival rates in vaccinated chickens challenged 7, 14, 21 or 31 d after vaccination were 0.47, 0.77, 0.97 and 0.92, respectively. All unvaccinated chickens died due to Newcastle disease (ND) following challenge. Chickens in Australia and Malaysia were bled and the serums tested for haemagglutination-inhibiting (HI) antibody to NDV. Many vaccinated birds with no detectable antibody, and all birds with a log2 titre of 2 or greater, survived challenge. The results showed that this V4 vaccine induced protective immunity in a significant proportion of chickens within 7 d of mass aerosol vaccination. This early immunity occurred in the absence of detectable circulating HI antibody. Non-HI antibody mediated immunity continued to provide protection up to 31 d after vaccination. Almost all vaccinated birds were protected within 3 w of vaccination. It is concluded that the V4 vaccine is efficacious and could be useful during an outbreak of virulent ND in Australia.