鸡新城疫消化道免疫的研究——Ⅰ.幼雏消化道免疫形态学及免疫功能研究

本实验采用消化道直接免疫法,阐述了其免疫机制及免疫效果的初步研究。用1日龄具有母源抗体的雏鸡,做Ⅱ系活苗嗉囊注射和饮水首免,20日龄Ⅰ系活苗嗉囊注射和饮水二免。通过对血清 HI 抗体效价连续检测及攻毒试验,结果证明幼雏早期消化道免疫后,HI 抗体效价维持在一定水平,对强毒攻击具有较高保护力。实验中,对雏鸡消化系统及免疫器官做了系统的免疫形态学观察。免疫后,肠粘膜增厚,固有膜淋巴样细胞、浆细胞及淋巴滤泡聚集、增生.提示采用消化道免疫途径可以诱导消化道局部免疫,并刺激全身淋巴组织产生免疫反应。     

NDV、IBDV二联灭活苗对蛋雏鸡免疫程序研究

为探讨鸡新城疫病毒(NDV)、传染性法氏囊病毒(IBDV)二联灭活疫苗在蛋雏鸡养殖中的实际应用效果进行本试验。用鸡新城疫、传染性法氏囊二联灭活疫苗及传染性法氏囊活疫苗以不同免疫程序免疫7~14日龄蛋雏鸡,定期监测NDV、IBDV抗体水平,并在雏鸡60日龄时进行攻毒保护试验。结果表明,各二联苗免疫组NDV抗体水平在免疫后14 d即达到NDV血清学最低有效保护价标准,并维持至雏鸡60日龄。免疫后28 d,灭活苗一次性免疫各组IBDV中和抗体水平显著高于活苗两次免疫组,琼扩抗体水平与活苗两次免疫组无明显差异。雏鸡60日龄时攻毒,各免疫组均100%保护,对照组无保护。以上结果说明该二联灭活疫苗可替代IBDV活毒疫苗用于7~14日龄商品雏鸡免疫,免疫期至少为60d。     

鸡新城疫首免采用饮水法的可行性探讨

监测雏鸡母源抗体水平以确定最佳免疫日龄,通过饮水给予不同剂量的新城疫疫苗,对雏鸡进行免疫,免疫后监测雏鸡的抗体水平,并用强毒流行株进行攻毒以检测保护率,探讨饮水免疫在雏鸡首免中应用的可行性。结果发现,雏鸡母源抗体在9日龄达峰值,14日龄时低于4(log2),饮水法效果较滴鼻点眼法差。因此,鸡新城疫首次免疫不提倡采用饮水法。     

鸡新城疫-传染性支气管炎-传染性法氏囊病三联灭活疫苗对不同日龄雏鸡的免疫效力研究

为评估鸡新城疫(ND)-传染性支气管炎(IB)-传染性法氏囊病(IBD)三联灭活疫苗对不同日龄和不同水平母源抗体雏鸡的免疫效力和持续期,本试验用该疫苗免疫7、14、21日龄SPF雏鸡和有母源抗体的普通雏鸡,免疫后采血测定ND血凝抑制抗体(HI Ab)、IB血凝抑制抗体(HI Ab)及IBD中和抗体(NA),并用传染性法氏囊病病毒(IBDV)强毒攻击。结果显示,7日龄SPF雏鸡免疫后21 d ND HI抗体、IB HI抗体及IBD中和抗体效价分别为7.9log2、6.9log2和14.1log2,SPF鸡日龄越大,抗体水平越高;28日龄SPF鸡免疫后3个月,0.3 mL免疫剂量组试验鸡ND HI、IB HI及IBD中和抗体效价分别达6.5log2、6.1log2和13.6log2,IBDV攻毒保护率均为100%(10/10);不同日龄普通雏鸡免疫效果与SPF鸡试验一致,抗体水平随鸡日龄增大而升高,IBD攻毒保护率也都达到100%(10/10)。试验结果证实,鸡新城疫-传染性支气管炎-传染性法氏囊病三联灭活疫苗可使7、14及21日龄SPF雏鸡和普通雏鸡产生良好的免疫力,对雏鸡的免疫期至少为3个月。     

对污染鸡场IBD免疫程序的确定

据雏鸡传染性法氏囊病(IBD)母源抗体消长情况,制订了污染鸡场IBD免疫程序。种鸡在雏鸡时用弱毒苗免疫两次,19和40周龄时用灭活苗再各免疫一次,其后代母源抗体在17日龄(伊莎褐)、18日龄(京白)阳性率降到50％,21～22日龄后基本消失,据此,制订了17～18日龄首次饮水免疫,26～27日龄再次免疫的免疫程序。通过对10万余只雏鸡试用,保护率达97.1％,较该场常规免疫程序(7～11日龄首免,17～21日龄再免)提高8.8％.     

堆型艾美耳球虫早熟株免疫雏鸡后血清抗体水平动态变化的研究

用堆型艾美耳球虫早熟株对雏鸡进行一次免疫和二次免疫试验 ,研究雏鸡血清抗体的动态变化。试验结果显示 ,在一次免疫试验中 ,各剂量组雏鸡均在免疫后 2天血清抗体出现较高的 OD值 ,之后呈下降趋势 ,11天后逐渐上升 ,到 12～ 13天时出现第 2个峰值 ,各组间的血清抗体 OD值比较 ,3 0 0 0个卵囊 /只剂量组极显著 (P<0 .0 1)高于 10 0 0个 /只、2 0 0 0个 /只和 5 0 0 0个 /只剂量组。在二次免疫试验中 ,8日龄首免、14日龄二免组雏鸡的抗体水平动态变化整体呈上升趋势 ,峰值 (0 .3 75 )出现在二免后第 10天 ,其后稳定地维持在较高水平 (平均 OD值为 0 .3 5 ) ,并且其抗体水平显著地高于 6日龄首免、12日龄二免组 (P<0 .0 5 )。抗体水平动态变化表明 ,堆型艾美耳球虫早熟株有较强的免疫保护力 ,其免疫方式可选用一次免疫 ,剂量为 3 0 0 0个卵囊 /只 ,也可采用二次免疫 ,即 8日龄首免 (15 0 0个 /只 ) ,14日龄二免 (15 0 0个 /只 ) ,从抗体动态变化的稳定性看 ,二次免疫更佳     

鸡新城疫－传染性支气管炎－传染性法氏囊病三联灭活疫苗对不同日龄雏鸡的免疫效力研究

为评估鸡新城疫（ND）-传染性支气管炎（IB）-传染性法氏囊病（IBD）三联灭活疫苗对不同日龄和不同水平母源抗体雏鸡的免疫效力和持续期,本试验用该疫苗免疫7、14、21日龄SPF雏鸡和有母源抗体的普通雏鸡,免疫后采血测定ND血凝抑制抗体（HI Ab）、IB血凝抑制抗体（HI Ab）及IBD中和抗体（NA）,并用传染性法氏囊病病毒（IBDV）强毒攻击。结果显示,7日龄SPF雏鸡免疫后21 d ND HI抗体、IB HI抗体及IBD中和抗体效价分别为7.9log2、6.9log2和14.1log2,SPF鸡日龄越大,抗体水平越高;28日龄SPF鸡免疫后3个月,0.3 mL免疫剂量组试验鸡ND HI、IB HI及IBD中和抗体效价分别达6.5log2、6.1log2和13.6log2,IBDV攻毒保护率均为100%（10/10）;不同日龄普通雏鸡免疫效果与SPF鸡试验一致,抗体水平随鸡日龄增大而升高,IBD攻毒保护率也都达到100%（10/10）。试验结果证实,鸡新城疫-传染性支气管炎-传染性法氏囊病三联灭活疫苗可使7、14及21日龄SPF雏鸡和普通雏鸡产生良好的免疫力,对雏鸡的免疫期至少为3个月。     

鸡新城疫-传染性法氏囊病-禽流感(H9亚型)三联灭活疫苗对雏鸡的免疫效力研究

为了评估鸡新城疫(ND)-传染性法氏囊病(IBD)-禽流感(AI)(H9亚型)三联灭活疫苗对雏鸡的免疫效力,采用该疫苗分别免疫1、7、10、14、21日龄SPF雏鸡和带有母源抗体的普通雏鸡,免疫后21d~28d采血测定ND血凝抑制抗体(HIAb)、IBD中和抗体(NA)及AI(H9)HIAb抗体,并用传染性法氏囊病病毒(IBDV)强毒攻击。结果显示,免疫后各不同日龄免疫组SPF雏鸡的ND HIAb、IBD NA及AI(H9)HIAb滴度分别为7.9log2~8.3log2、13.9log2~15.8log2及8.0log2~8.9log2,IBDV攻毒均100%保护;而普通雏鸡免疫后的ND HIAb、IBD NA及AI(H9)HIAb滴度,1日龄免疫组分别为5.9log2、12.1log2和5.6log2,7、10、14、21日龄免疫组分别为6.1log2~7.0log2、12.6log2~14.5log2和6.0log2~7.5log2;IBDV攻毒,1日龄免疫组普通雏鸡保护率为60%,7、14及21日龄免疫组保护率均为100%;结果证实,SPF雏鸡所产生的免疫效力要高于带有母源抗体的普通雏鸡,1日龄普通雏鸡免疫所产生的免疫效力明显低于其他日龄。根据试验结果,推荐该疫苗对雏鸡的使用日龄为7日龄~14日龄。     

鸡新城疫免疫程序的试验

多年来,我省对出壳1、2日龄雏鸡用Ⅱ系苗点鼻,秋防用Ⅰ系苗注射或刺种的新城疫免疫程序。近年来,自提出鸡新城疫母源抗体能干扰主动免疫的问题之后,已改为孵出后7～10日龄用Ⅱ系苗作首次免疫,25～30日龄时用Ⅱ系苗作第二次免疫,两个月龄     

通过免疫对比试验解决一起防疫纠纷的体会

某养殖户饲养蛋鸡500羽．雏鸡、饲料及疫苗均有雏鸡供应方提供。该批鸡的免疫程序为10日龄时．用鸡新城疫IV系苗及IBD苗．1倍量饮水；25日龄时，用鸡新城疫IV系苗，1倍量饮水；42日龄时，养殖户使用兽医站提供的鸡新城疫I系苗．1倍剂量，腿部肌肉免疫注射。免疫后的第2天．鸡群的采食量明显下降．精神萎靡不振．发生少量死亡。     

鸡葡萄球菌病发病机理的研究

本实验采用多种途径感染,系统地研究了鸡葡萄球菌病的发病机制、菌血症出现的时间、细菌在体内的分布,并对病变模式作了详细观察。结果表明,静脉、肌肉和关节途径最敏感,皮下、呼吸道、腹腔和眼睛途径次之,而消化道不敏感。细菌在体内主要分布于肝脏、脾脏、其次在肾、肺和心脏。静脉接种鸡只持续出现菌血症,皮下、腹腔、气管、眼睛和消化道途径均在感染12小时后出现菌血症,而肌肉和关节途径分别在6和24小时后出现。除消化道途径外,其它七种途径均可形成不同程度的典型病理变化,静脉、肌肉和关节途径感染可导致骨髓炎和关节炎的发生。组织学观察表明,鸡葡萄球菌病对免疫器官的发育和功能有一定的抑制和破坏作用。     

网状内皮增殖病病毒(REV)感染SPF雏鸡后局部免疫组织抗体生成细胞的变化

对 SPF雏鸡人工感染 REV后局部免疫组织盲肠扁桃体和哈德氏腺的 Ig A、Ig G和 Ig M抗体生成细胞数量动态变化进行了检测。结果发现 ,REV感染 SPF雏鸡后 7～ 4 9天消化道局部免疫组织盲肠扁桃体和呼吸道相关局部免疫组织哈德氏腺的 Ig A、Ig G和 Ig M抗体生成细胞数量明显低于未感染对照雏鸡。表明雏鸡感染 REV后消化道局部组织盲肠扁桃体和呼吸道相关免疫组织哈德氏腺体液免疫机能均显著下降。     

一定纯度中药复方多糖对鸡细胞免疫和体液免疫的影响

为筛选对鼠免疫增强效果显著的一定纯度中药复方多糖(cCHMPS)对雏鸡的最佳免疫增强剂量,本试验将300只1日龄雄性良凤青脚麻鸡随机分为6组,即空白对照组、盐酸左旋咪唑组、黄芪多糖组及cCHMPS高、中、低剂量组,每组各50只.分别于8日龄皮下注射生理盐水、盐酸左旋咪唑、黄芪多糖及不同剂量的cCHMPS,连续7 d,在免疫后第8、14、21、35、42天采血,测定外周血中T、B淋巴细胞增殖活性及血清中新城疫病毒抗体含量.试验结果表明,黄芪多糖和不同剂量cCHMPS均能显著或极显著促进外周血T、B淋巴细胞增殖(P<0.05;P<0.01),提高新城疫病毒抗体浓度,增强机体细胞免疫和体液免疫水平,且中剂量的cCHMPS的免疫增强效果最好.     

抗鸡新城疫和传染性法氏囊病病毒联合高免卵黄抗体的保护率测定

应用抗鸡新城疫病毒(NDV)和传染性法氏囊病病毒(IBDV)联合高免卵黄抗体(NDV-HI效价1:640以上,IBDV琼扩效价1:80以上)肌注免疫,可抵抗ND或IBD强毒对1～2月龄雏鸡的攻击;饮水免疫效果不确实.体内抗体完全消失的鸡发生ND后,用卵黄抗体难以治愈,体内抗体未完全消失的鸡发生ND后,卵黄抗体的治愈率很高.临床应用总有效率在90%以上.     

鸡新城疫重组鸡痘病毒活疫苗的免疫持续期和加强免疫研究

本研究旨在评价表达新城疫病毒（NDV）血凝素-神经氨酸酶（HN）基因的重组鸡痘病毒（rFPV-12LSHN）活疫苗的免疫持续期和加强免疫对疫苗免疫效力的影响。用rFPV-12LSHN活疫苗免疫14日龄SPF鸡,103PFU/羽,7d即可检测到NDV HI抗体应答,对NDV强毒F48E8株攻毒保护率达100%。一次免疫18周后,对NDV强毒攻击依然提供完全保护。鸡痘病毒（FPV）疫苗免疫4周,再接种rFPV-12LSHN活疫苗,攻毒保护率降低至50%。相反,rFPV-12LSHN免疫4周,随后二次免疫可显著提高对NDV的体液免疫应答水平（P〈0.01）,对NDV强毒攻击的保护率仍然为100%。结果表明,表达NDV HN基因的重组鸡痘病毒（rFPV-12LSHN）活疫苗,能够快速建立坚强免疫力,免疫持续期至少可达18周,rFPV-12LSHN的二次免疫可以提高疫苗的免疫力。     

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.     

怀牛膝多糖对雏鸡疫苗免疫效果的影响

150羽14日龄蛋公鸡随机均分成3组,用新支二联弱毒苗点眼滴鼻免疫的同时,2个试验组分别肌肉注射高、低剂量的怀牛膝多糖溶液,对照组注射生理盐水,每天注射1次,连续注射3次。分别于免疫后第7、14、21、28、35和42天各组随机抽取8只鸡翼下静脉采血,分离血清,用微量法测定新城疫血凝抑制（HI）抗体效价,于免疫后第10、20、30、40和50天每组随机抽取5只鸡心脏采血,分离淋巴细胞,用MTT法测定淋巴细胞增殖的动态变化,流式细胞仪双染色法检测CD^4＋、CD^8＋T淋巴细胞含量,并放血致死称取体重和胸腺、脾脏、法氏囊的质量,计算器官指数。结果表明,怀牛膝多糖高剂量组的HI抗体效价、CD^4＋/CD^8＋比值和免疫器官指数都明显高于对照组,并且高剂量怀牛膝多糖可以显著促进T淋巴细胞增殖。提示怀牛膝多糖对鸡新支二联疫苗的免疫效果有明显的增强作用。     

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.     

不同感染量REV对鸡免疫反应和细胞毒性作用的影响

用不同剂量网状内皮增生症病毒（REV）感染肉鸡和SPF鸡后，检测血液中T淋巴细胞对ConA的反应和NK细胞、细胞毒T细胞（CTL）的细胞毒性作用以及NDV抗体生成变化等，观察REV感染对机体非特异性、特异性细胞免疫反应和体液免疫反应的影响。结果表明无论高剂量还是低剂量REV感染均造成体液免疫和非特异性细胞免疫抑制，而且高剂量比低剂量对免疫功能的抑制作用更强，但对NK细胞和细胞毒T细胞的细胞杀伤活性却有升高趋势，在抗肿瘤方面发挥一定的作用。这种结果说明REV感染对机体的免疫抑制是有选择性的，且抑制程度与感染病毒量有关。     

ADMINISTRATION OF A VACCINE PREPARED FROM THE AUSTRALIAN V4 STRAIN OF NEWCASTLE DISEASE VIRUS BY AEROSOL AND DRINKING WATER

SUMMARY An experimental vaccine containing the avirulent Australian V4 strain of Newcastle disease virus was used to vaccinate 3- or 6-week-old chickens by aerosol and drinking water application. The chickens lacked maternally derived antibody to Newcastle disease virus. When the vaccine virus was diluted in tap water more than 90% of the infectivity was destroyed immediately. The addition of 0.25% skim milk prevented this loss and there was no loss in distilled water. Rates of inactivation at 37°C were similar in tap water and distilled water and were unaffected by the addition of skim milk. Both methods of vaccination resulted in the production of haemagglutination-inhibition antibodies which persisted for at least 8 to 12 weeks. The antibody response to aerosol vaccination was significantly better than that following drinking water vaccination. No clinical disease was induced by exposure to vaccine virus. Serum neutralisation antibodies paralleled those detected by haemagglutination-inhibition in chicks vaccinated once by drinking water. After revaccination through the drinking water, haemagglutination-inhibition antibodies were boosted temporarily while neutralising antibodies were maintained at an enhanced level. From chickens vaccinated by aerosol, Newcastle disease virus was recovered for 10 days from lungs and for 7 days from tracheas and caecal tonsils. Peak viraemia was detected 2 and 3 days after vaccination while both neutralising and haemagglutination-inhibition antibodies became detectable 5 days after vaccination.