鸽接种鸡痘病毒体内抗体消长规律的研究

为检测免疫鸡痘病毒(FPV)的幼鸽体内抗体消长规律和毒性,将试验鸽分成4组,按一个使用剂量、增加免疫剂量、二次免疫和对照组对幼鸽进行刺种接种.试验组接种鸡痘病毒,对照组接种生理盐水.在试验期间鸽子精神状态良好,采食、饮水正常,未观察到病理变化.在免疫后1～8周采血,分离血清,进行抗FPV特异性抗体和中和抗体效价检测.免疫后1周中和抗体开始升高,3周达到高峰,4周开始下降,8周仍高于对照组;增加免疫剂量组和二次免疫组产生的抗体水平高,但消长规律与使用剂量组相同.结果表明鸡痘病毒能刺激鸽产生特异性抗体和中和抗体,且维持时间较长,无毒性,生物安全性好.     

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。     

鸡传染性法氏囊病病毒超强株VP2基因重组新城疫病毒胚胎免疫安全性及效力试验

为研制能够同时预防传染性法氏囊病(IBD)和新城疫(ND)的疫苗,本研究选用表达IBD病毒(IBDV)超强毒株(vvIBDV)VP2基因的重组ND病毒(NDV)LaSota疫苗株(rL-VP2),测定其半数鸡胚感染量、平均鸡胚致死时间、脑内接种指数和静脉内致病指数等指标,按不同剂量接种18胚龄SPF鸡胚,分别于出雏后第9d、第14d和第21d采血,用微量凝集法和ELISA方法测定血清中抗NDV抗体和抗IBDV抗体水平,并于出雏后28d用NDV强毒(F48E9株)和vvIBDVGx株攻毒,评估重组疫苗的胚胎免疫效果。结果显示:按104EID50/枚剂量进行免疫不会影响SPF鸡胚出雏率和出雏后21d存活率,并且该免疫组雏鸡能够对NDV强毒和vvIBDV攻毒提供安全的免疫保护。     

CpG ODN对传染性腔上囊病病毒活疫苗免疫效果的影响

将120只来航系SPF鸡随机分成对照组、注射CpG ODN组、IBDV活疫苗点眼免疫组及IBDV活疫苗 CpG ODN免疫组，每组30只，于7日龄时对试验鸡进行初次免疫，21日龄时进行加强免疫，28日龄时用IBDV野毒株攻毒。检测不同时间的IBDV特异性抗体效价、血液和脾淋巴细胞的Con A刺激指数(SI)。结果表明，CpG ODN处理后脾细胞的增殖转化能力强于血液淋巴细胞的增殖转化能力；CpG ODN的使用使血清中IBDV特异性抗体产生时间提前7d，抗体效价升高；与对照组相比，仅用CpG ODN处理就可使1／3的鸡攻毒后迅速产生高效价(约1：3000)的特异性抗体，降低了发病率和死亡率，证明CpG ODN能增强IBDV活疫苗的免疫效果。     

鸡传染性支气管炎病毒与新城疫病毒增殖作用及免疫干扰的研究

试验旨在观察鸡传染性支气管炎病毒（infectious bronchitis virus,IBV）和鸡新城疫病毒（newcastle disease virus,NDV）之间的增殖干扰现象,分析在免疫过程中两种疫苗存在的免疫干扰,为确定疫苗的免疫程序提供依据。采用完全随机试验设计,选取10日龄鸡胚尿囊腔接种不同浓度的IBV、NDV以及不同浓度混合的IBV和NDV,收集尿囊液测定其血凝效价（HA）;用不同浓度的IBV、NDV以及不同浓度混合的IBV和NDV分别免疫BALB/c小鼠及SPF雏鸡,收集血清,间接酶联免疫吸附法（ELISA）和血凝抑制（HI）试验测定IBV和NDV的抗体效价及抑制效价。结果表明,同胚培养时,无论先接种IBV后接种NDV还是先接种NDV后接种IBV或IBV和NDV同时接种,IBV对NDV均有干扰作用,而NDV对IBV没有干扰作用;接种IBV和NDV的小鼠,其产生IBV和NDV的抗体效价均低于单独免疫组,免疫的次序及免疫间隔时间对IBV和NDV的抗体效价均有影响,IBV对NDV的免疫效果具有较大的干扰作用。不同针混合免疫方式能提高IBV和NDV的抗体效价;接种IBV和NDV的雏鸡,免疫次序及免疫相隔时间对IBV和NDV的抗体效价均有影响,IBV对NDV的免疫效果具有较大的干扰作用;雏鸡和小鼠免疫血清HI试验数据表明,免疫次序及免疫间隔时间对IBV和NDV抗体的产生均有影响,但对NDV抗体的产生影响更明显,随着免疫间隔时间的增加,IBV和NDV的抗体水平呈增加趋势,雏鸡比小鼠更能敏感地反映出IBV对NDV的免疫干扰作用。混合注射时IBV和NDV的抗体水平均降低,IBV对NDV的免疫效果有干扰作用。因此,在同胚培养、小鼠及雏鸡的免疫中,IBV对NDV有干扰作用,而NDV对IBV无干扰作用。混合接种时IBV和NDV抗体效价均下降,IBV对NDV的免疫效果有干扰作用。免疫次序及免疫间隔时间对IBV和NDV抗体的产生均有影响,但随着免疫间隔的增加,IBV和NDV的抗体水平呈增加趋势。     

猪繁殖与呼吸综合征对塞内卡病毒灭活苗免疫效果的影响

为探究猪繁殖与呼吸综合征(PRRS)对塞内卡病毒(SVA)灭活疫苗免疫效果的影响,对PRRS阳性猪和PRRS阴性猪分别免疫相同批次、相同剂量的SVA灭活苗,检测首次免疫后7 d、14 d、21 d、28 d血清中和抗体效价,并在免疫后28 d对所有试验猪进行攻毒保护试验。结果显示,所有PRRS阳性猪的SVA血清中和抗体效价均小于1∶4,而PRRS阴性猪SVA血清中和抗体效价在二免后上升到较高水平。攻毒保护结果与中和抗体效价结果呈正相关,PRRS阳性组0/5保护,而PRRS阴性组试验猪达到5/5保护。这一研究表明PRRS会使SVA灭活疫苗免疫效果下降,为SVA及PRRS疫苗的研发及猪场疾病的防控提供了参考。     

兔自家大肠杆菌高免卵黄抗体的研制与应用

采用灭活菌苗进行基础免疫、加强免疫,灭活油乳剂疫苗进行超强化免疫和维持免疫程序。通过微量凝集试验方法,研究了鸡抗兔大肠杆菌高免卵黄抗体的消长规律及临床应用。结果表明,超强化免疫5周后,抗体效价达到高峰,9周后抗体效价逐渐下降。同一剂量卵黄抗体效价达26时,肌注2 mL/kg体重,连用3 d对兔大肠杆菌感染的治愈率达90%。成品卵黄抗体的效价应达到26、安全性检验、无菌检验阴性方可用于临床治疗。卵黄抗体在血清中维持高浓度可持续1周,然后逐渐下降。在冷冻环境下可保存1年,其抗体效价保持在有效水平。与头孢噻肟进行的对比田间治疗试验,卵黄抗体的治愈率显著高于头孢噻肟。     

甲硝唑对鸡传染性法氏囊病活疫苗免疫效果的影响

本研究观察了药物对传染性法氏囊病(IBD)活疫苗初次免疫和二次免疫的影响,结果显示高抗传染性法氏囊病病毒(IBDV)母源抗体雏鸡IBD活疫苗初次免疫前应用10～100毫克/千克甲硝唑和10毫克/千克左旋咪唑,在免疫鸡血清抗IBDV抗体水平均明显下降,甲硝唑用药鸡疫苗二次免疫后,血清抗IBDV抗体滴度明显高于未用药对照组.10毫克/千克左旋咪唑给药组疫苗二次免疫后鸡血清抗IBDV抗体滴度与未用药对照组无明显差别.     

四种雉鸡新城疫疫苗免疫抗体消长规律的研究

用新城疫活疫苗(La Sota株)和新城疫、传染性支气管炎二联灭活疫苗免疫动物园圈养的孔雀、七彩山鸡、原鸡和白冠长尾雉4种雉鸡,于免疫前及免疫后14、21、35、90、150、210、280d分别采血检测血清中新城疫血凝抑制(HI)抗体。结果表明,孔雀免疫接种新城疫疫苗后14d就能产生较高效价HI抗体,35d抗体效价达高峰;七彩山鸡、原鸡、白冠长尾雉免疫接种新城疫疫苗后14d也能产生较高效价抗体,21d抗体水平达高峰,至免疫接种后90d抗体效价逐渐降低。整个试验过程中,孔雀产生的HI抗体效价最高,并且整齐度好,维持时间最长,至免疫接种后280d抗体效价仍能维持在7.5log2;白冠长尾雉产生HI抗体效价次之,到免疫接种后280d仍维持在6.67log2;七彩山鸡和原鸡抗体下降较快,到免疫接种后280d分别下降到5.67log和4.71log2。     

蛋用型和肉用型雏鸡抗新城疫病毒母源抗体半衰期测定

分别于不同日龄采集海兰褐蛋用型雄性雏鸡和AA肉用型雏鸡血清,测定其对新城疫病毒(NDV)HI抗体水平,以此确定抗NDV母源抗体的衰减规律及半衰期。结果表明,雏鸡出壳后抗体水平持续升高,到4日龄左右达到最高峰,随后快速下降,三周后抗体几乎消失。蛋用型雏鸡血清中对NDV母源抗体的半衰期为2.19d左右。肉用型雏鸡血清中对NDV母源抗体的半衰期为3.10d左右。     

Effect of porcine circovirus type 2 (PCV2) vaccination on porcine reproductive and respiratory syndrome virus (PRRSV) and PCV2 coinfection

The objectives were to determine if PCV2 vaccination is effective in reducing disease and lesions associated with PRRSV and PCV2 coinfection and if there is a difference between intradermal (ID) and intramuscular (IM) route of PCV2 vaccination. Seventy-four, 21-day-old pigs were randomly allocated into one of six groups. On day 0, pigs were vaccinated with 2ml Suvaxyn((R)) PCV2 One Dose (Fort Dodge Animal Health, Inc.) by intramuscular (VAC-M-COINF) or intradermal (VAC-D-COINF) routes. On day 28, pigs were either singularly (PRRSV-only, PCV2-only) or coinfected (COINF) with PRRSV and PCV2. All pigs in all groups were necropsied on day 42. All vaccinated pigs seroconverted (IgM, IgG, and neutralizing antibodies) to PCV2 between 14 and 28 days post-vaccination. After challenge, all groups inoculated with PRRSV had reduced average daily gain compared to CONTROLS and PCV2-only (P<0.001). COINF pigs had significantly (P<0.05) reduced anti-PCV2-IgG antibody levels and neutralizing antibody levels compared to both vaccinated groups. COINF pigs had more severe lung lesions compared to VAC-M-COINF (P<0.05). COINF pigs had higher amounts of PCV2 DNA in serum samples and feces (P<0.05) and increased amounts of PCV2 in lymphoid tissues (P<0.05) compared to both vaccinated groups. In summary, PCV2 vaccination was effective at inducing a neutralizing antibody response and significantly reducing PCV2-associated lesions and PCV2 viremia in pigs coinfected with PCV2 and PRRSV. Differences between intradermal and intramuscular routes of vaccine administration were not observed.     

PRRSV ORF6及IL-18基因重组真核质粒试验免疫研究

将本实验室构建的重组质粒接种PRRSV的自然宿主断奶仔猪,分析其诱发细胞免疫和体液免疫应答的能力。结果表明：重组质粒pEGFP-ORF6和pEGFP-IL18-ORF6免疫组在首免后14d开始产生特异性抗体,且其产生抗体水平随时间呈上升趋势,但pEGFP-ORF6和pEGFP-IL18-ORF6两组之间产生抗体差异不显著（P〉0.05）。中和抗体检测结果显示只有pEGFP-ORF6在首免后42d有低水平的中和抗体产生。细胞免疫检测结果表明,pEGFP-IL18-ORF6诱导T淋巴细胞增殖和促进IFN-γ和IL-2的分泌的能力显著高于pEGFP-ORF6和pEGFP-IL18（P〈0.05）,表明IL-18有效地发挥了分子其佐剂的效用,能增强细胞免疫应答并介导较好的Th1类免疫反应。     

Induction of systemic immune responses in sheep by topical application of cholera toxin to skin

Cholera (and related) toxins (CT) when applied topically on unbroken skin induce systemic immune responses in mice, a procedure called transcutaneous immunization (TCI). The current study examined the capacity for TCI to induce systemic immune responses in sheep. Three groups (n=5 per group) were immunized at day 0 (priming) and day 28 (boosting) with 250 microg of CT in water by TCI, with 25 microg of CT in alum by intramuscular injection, or not immunized. Serum samples were taken at days 0, 28, 42, 56 and 70 after immunization for measurement of CT-specific IgG as well as CT-specific IgG1, IgG2, IgA and IgM antibodies by ELISA. After immunization, IgG, IgG1 and IgG2 antibody in immunized groups were significantly higher than in the control group, and boosting further increased these titres. IgG, IgG1 and IgG2 in the injection group were significantly higher than in the TCI group. There was a preponderance of IgG1 antibody, relative to IgG2, in both immunized groups. CT-specific IgA and IgM were detected in both immunized groups. Lymphocyte proliferation to CT was measured at day 90. A CT-specific lymphocyte proliferative response (stimulation index>2) was detected in all sheep from the injection group, in two sheep from the TCI group and in none of the controls. Results demonstrated that TCI induces primary and secondary antibody responses and specific proliferative responses to CT in sheep.     

不同纯化浓缩方法制备的新城疫疫苗对SPF鸡免疫效果的研究

为研究利用不同纯化浓缩方法制备的新城疫疫苗对SPF鸡免疫效果的影响,试验取经过纯化浓缩的新城疫抗原制备的疫苗、经离心未经浓缩的抗原制备成的疫苗和经浓缩未离心的抗原制备成的疫苗分别免疫30日龄SPF鸡,免疫后分别在第7d、14 d、21 d、28 d、35 d、45 d、70 d、90 d采集血清检测新城疫抗体水平。结果表明,经浓缩纯化后抗原制备的疫苗与其他两组抗原制备的疫苗相比,物理外观性状基本相同,浓缩纯化后抗原制备的疫苗其单位抗原含量更高、杂质更少,抗体水平更高,统计数据显示差异显著。因此研究得出,浓缩倍数(即抗原含量)越高,抗体水平越高,持续的时间也越长。     

鸡病毒性关节炎弱毒苗的研制 Ⅱ.弱毒株的免疫原性

应用具有良好安全性的鸡病毒性关节炎病毒（AVAV）弱毒JN-1株免疫1日龄肉鸡40只,分别用琼脂扩散试验、间接ELISA和中和试验（NT）检测免疫鸡血清中的AVAV抗体。中和抗体于免疫后7d开始出现,14d后全部阳转;ELISA抗体也于免疫后7d开始显示阳性,21d后全部阳转;琼脂扩散抗体于免疫后14d开始阳转,28d以后全部显示阳性。分别在免疫后7、14、21和 28d于足掌皮下用强毒攻击免疫鸡,临床保护率分别为2/10、9/10、10/ 10、10/10,病理保护率分别为0/10、7/10、9/10、10/10。体外抗病毒谱试验显示,弱毒株的免疫血清能中和AVAV J-1株、S_(1133)株、FDO株和H_(9307)株。上述实验结果表明,AVAV弱毒株具有良好的免疫原性。     

鸡新城疫-禽流感二联灭活疫苗免疫不同日龄商品肉鸡的效果比较

使用鸡新城疫-禽流感（H9N2 HP株）二联灭活疫苗分别免疫3、7、14日龄三组商品肉鸡各40羽,同时设一组空白对照组。各免疫组及对照组于3、7、14、21、28、35、42日龄采血检测新城疫、禽流感抗体,于21、28、35日龄进行禽流感病毒攻毒,对比不同日龄免疫组的抗体消涨情况及不同日龄禽流感攻毒结果。发现对照组随鸡日龄增加,新城疫与禽流感抗体逐渐下降,在42日龄时下降至0,而不同免疫组新城疫与禽流感抗体均先下降,21日龄左右开始上升,至35日龄新城疫与禽流感抗体升至6log2以上。3日龄免疫组的禽流感免疫保护效果最好,21、28、35日龄时禽流感强毒攻毒保护率均达100%;7日龄免疫组在21、35日龄时禽流感强毒攻毒保护率均达100%,28日龄时禽流感强毒攻毒保护率达70%;14日龄免疫组在28、35日龄时禽流感强毒攻毒保护率均达100%,21日龄时禽流感强毒攻毒保护率只达30%。试验表明,商品肉鸡选择3日龄免疫鸡新城疫-禽流感（H9N2 HP株）二联灭活疫苗时禽流感免疫保护效果最好,采用3日龄免疫程序可以提高新城疫与禽流感的免疫保护效果,减少养殖业的经济损失。     

怀山药多糖对鸡免疫功能的影响

目的研究怀山药多糖对鸡免疫功能的影响。方法将150羽14日龄蛋公鸡随机分成3组,每组50羽,14日龄用一羽份新支二联弱毒苗点眼滴鼻免疫;同时2个试验组分别肌肉注射怀山药多糖溶液高、低剂量各0.3ml/羽,对照组注射生理盐水0.3ml/羽,连用3d;分别于免疫用药后的第7、14、21、28、35、42天各组随机抽取8只翼下静脉采血,后分离血清,测定新城疫血凝抑制(HI)抗体效价,并于免疫后14、21、28、35、42天随机抽4只放血致死后称重,摘取胸腺、法氏囊、脾脏称重并计算指数。结果试验组特异性免疫抗体和免疫器官指数均高于对照组,在某些时间点差异显著。结论怀山药多糖能增强鸡的免疫功能,可作为免疫增强剂与疫苗同时使用。     

Immune response in skunks to a vaccinia virus recombinant expressing the rabies virus glycoprotein.

Striped skunks (Mephitis mephitis) were vaccinated with a vaccinia virus recombinant expressing the rabies virus glycoprotein. Virus neutralizing antibodies to rabies virus were present at 14 days postvaccination by the following routes: scarification (6/6), intramuscular (4/4) and intestinal (5/8). Six out of seven skunks that ate vaccine filled baits had virus neutralizing antibodies at 28 days. When challenged intramuscularly with street virus, the survival rates were 5/7 for the bait-fed group, 4/8 for the intestinal group, 3/4 for the intramuscular group, 5/6 for the animals that were scarified, and 0/8 for controls. This is the first report of a high rate of immunization of skunks with a rabies vaccine administered orally.     

猪乳中抗猪瘟IgA、IgG、IgM抗体动态变化规律研究

以猪瘟弱毒疫苗免疫空怀母猪,待母猪分娩后,分别收集10 d内的乳汁,以建立的间接ELISA方法检测猪瘟IgA、IgG、IgM水平,并观察各种抗体动态变化规律。试验结果表明,猪初乳中的IgA、IgG、IgM抗体效价均在分娩当天达到最高值,随后迅速下降,7 d后抗体水平与常乳基本一致。对照组母猪仅在分娩当天检测到较低的抗体水平。     

Immunization of pigs with a purified Shiga-like toxin II variant toxoid.

Passive transfer of neutralizing antibodies and active immunization with a toxoid of purified Shiga-like toxin II variant (SLT-IIv, edema disease toxin) were used to protect pigs against challenge with SLT-IIv. Six 6-week-old pigs were passively immunized by intraperitoneal administration of an immunoglobulin preparation from porcine antiserum against purified SLT-IIv. Six 6-week-old pigs and twelve 2-week-old pigs were actively immunized by two intramuscular injections of 25 micrograms of SLT-IIv toxoid given 2 weeks apart. The 24 immunized pigs and an equal number of age-matched unimmunized control pigs were all challenged by intravenous injection of purified SLT-IIv (6 ng/kg body weight). The six passively immunized pigs acquired neutralizing SLT-IIv antibody titers of 1280 or 2560 and the 18 actively immunized pigs mounted a serum immune response with toxin neutralizing titers ranging from 128 to 10,240. All 24 immunized pigs survived the challenge but nine pigs with the lowest titers showed mild clinical signs of edema disease. All 24 control pigs died within 30 hours of challenge.