梅花鹿源牛病毒性腹泻病毒分离株油剂灭活苗的制备和免疫效果观察

为了探索有效防制鹿黏膜病的方法,对从吉林省长春市双阳区梅花鹿流产胎儿肝脏病料中分离出的牛病毒性腹泻病毒灭活后制备成油剂灭活苗,免疫接种试验动物后,检测其体液免疫和细胞免疫水平,结果表明,梅花鹿源牛病毒性腹泻病毒(BVDV)分离株油剂灭活苗既能产生特异性体液免疫,又可以产生细胞免疫应答。     

牛流行热弱毒疫苗的免疫试验

牛流行热弱毒疫苗经二年实验室和现地大面积区域试验结果表明:该疫苗是安全、有效的,在保存期和免疫期方面又明显优于灭活苗,因此是一种很有发展前途的疫苗。     

牛防疫程序一览

<正>1口蹄疫免疫程序一般采用春秋两季免疫。犊牛:出生后4~5个月首免,肌注牛羊A型口蹄疫灭活疫苗(单价苗)1毫升/头以及牛羊O型-亚洲I型口蹄疫双价灭活苗(多价苗)1毫升/头;首免后6个月二免(方法、剂量同首免)。青年牛、后备牛、成母牛:每年接种疫苗2次,每间隔6个月免疫一次,肌注牛羊A型口蹄疫灭活疫苗以及牛羊O型-亚洲I型口蹄疫双价灭活苗各2毫升/头。     

不同免疫程序对猪流行性腹泻疫苗免疫效果的影响

为了研究不同免疫程序对猪流行性腹泻疫苗免疫效果的影响,试验选取河南省8个猪场,每个猪场受试猪随机分为2组,采用2次猪腹泻二联灭活苗、2次猪腹泻三联活疫苗、1次猪腹泻二联灭活苗+1次猪腹泻三联活疫苗、2次猪腹泻三联活疫苗+1次猪腹泻二联灭活苗等4种免疫程序进行试验,母猪于产仔当天收集初乳样本942份,采用ELISA法检测猪流行性腹泻(PED) IgA抗体。结果显示,2次猪腹泻三联活疫苗+1次猪腹泻二联灭活苗的免疫效果最好,但成本相对较高,其次是2次猪腹泻三联活疫苗和1次猪腹泻二联灭活苗+1次猪腹泻三联活疫苗,最后是2次猪腹泻二联灭活苗。结果表明,猪腹泻三联活疫苗单独或与猪腹泻二联灭活苗组合使用效果优于猪腹泻二联灭活苗。     

猪流行性腹泻疫苗不同免疫程序的效果评估

为了探讨不同免疫程序对猪流行性腹泻疫苗免疫效果的影响,本试验选取猪流行性腹泻病毒(PEDV)抗原、抗体双阴的怀孕母猪,随机分为5个组,每组25头,分别给予2次猪腹泻二联灭活苗、2次猪腹泻二联活苗、1次猪腹泻二联活苗+1次猪腹泻二联灭活苗、2次猪腹泻二联活苗+1次猪腹泻二联灭活苗和疫苗稀释液(对照组)免疫。母猪产仔当天收集初乳样本,采用ELISA法检测PEDV的IgA抗体,筛出优秀免疫组,同时进行母源抗体持续时间检测。结果显示：2次猪腹泻二联活苗+1次猪腹泻二联灭活苗免疫后抗体整齐度最好,乳汁中IgA抗体阳性率为97.5%,IgA抗体持续时间达12d以上;2次猪腹泻二联活苗免疫后乳汁中IgA抗体阳性率为93.75%,效果优于1次猪腹泻二联活苗+1次猪腹泻二联灭活苗免疫;2次猪腹泻二联灭活苗免疫后乳汁中IgA抗体阳性率为38.13%,抗体水平最差。结果表明,猪腹泻二联活苗单独2次免疫或与猪腹泻二联灭活苗组合使用效果优于猪腹泻二联灭活苗单独使用。     

牛流行热病预防措施的研究

建立流行热监测制度．每年在流行季节前一个月对检测牛群按5％抽样检测流行热血清中和抗体。以流行热自然发病的抗体临界效价1：32^x为标准，将监测数据进行统计分析，根据分析结果．制订相应防治措施。 根据生长发育不同阶段．免疫用苗的特点，流行季节的长短和流行热自然发病的临界效价，建立合理的不同月龄段牛只的免疫程式。 选择切实安全有效的疫苗，是解决流行热免疫的基本条件，哈尔滨兽医研究所的灭活苗和澳大利亚Websters弱毒疫苗．都可以对牛提供有效的保护免疫。尤以先注射澳大利亚的弱毒苗后，再注射哈尔滨的灭活苗．效果最好．单独应用灭活苗次之，弱毒苗再次之，而先注灭活苗后再注弱毒苗效果最差。 流行热的发生和流行由相应的吸血昆虫传播．建立相应的防疫卫生制度，搞好环境卫生，定期进行药物杀虫或灯诱杀虫，在相应吸血昆虫活动的高峰期，改变收、放牧时间，减少牛只被传播昆虫叮咬。 上述技术的综合应用．我们在广州地区1991年流行热大流行时，成功地建立了一个无流行热病牛发生的奶牛场。     

牛流行热病毒灭活疫苗免疫后牛外周血淋巴细胞亚类分布的研究

对牛流行热病毒灭活疫苗免疫并攻每后，牛外周血淋巴细胞亚类的变化进行了研究。结果表明，灭活苗免疫后，牛的CD4^ 显著升高，可能与参与辅助B淋巴细胞合成抗体有关，攻毒后，γδT细胞均显著上升，免疫组在攻毒后3周仍保持在相当的高水平，IL－2Rα阳性细胞在攻毒后高热期也有显著升高，而CD8^ 细胞没有明显变化。     

山东省牛“猝死病”病原学研究

采用流行病学调查、毒物检测及微生物学检验等方法，对山东省牛“猝死病”发病原因进行了系统研究。结果证明，本病是由凝结芽胞杆菌和克雷伯氏杆菌肺炎亚种协同感染所致，并通过研制成的二联灭活菌苗，有效地控制了牛“猝死病”的发生。     

某地区牛、羊口蹄疫免疫抗体监测与分析

为确实掌握某地区牛、羊口蹄疫(FMD)春季防疫免疫效果,利用酶联免疫吸附试验(ELISA)对该地区某奶牛场的74头奶牛进行了A、O和亚洲Ⅰ型FMD免疫抗体的检测,对7家规模养殖场和7家散养户的80只山羊进行了O型和亚洲Ⅰ型FMD抗体的检测。结果显示,用O型-A型-亚洲Ⅰ型FMD三价灭活苗免疫奶牛后,可同时产生针对A、O和亚Ⅰ型3种亚型的高水平抗体;用O型-亚洲Ⅰ型FMD二价灭活苗免疫羊后,规模羊场的两种亚型抗体合格率均高于散养羊。所抽检牛、羊抗体合格率均超过农业部规定的70%的标准。结果表明,FMD三价苗免疫效果较好,可以起到一针防三型的作用,值得推广应用。     

牛溶血性曼氏杆菌及牛荚膜A型多杀性巴氏杆菌灭活疫苗对小鼠的保护性研究

牛溶血性曼氏杆菌及牛荚膜A型多杀性巴氏杆菌是导致牛呼吸道疾病（bovine respiratory disease,BRD）的重要细菌性病原,每年给养牛业带来巨大的经济损失,目前对其疫苗研究仍显不足。本研究选用牛溶血性曼氏杆菌（Mannheimia haemolytica,Mh） Mh422株和牛荚膜A型多杀性巴氏杆菌（Pasteurella multocida,Pm） PmCQ2株作为疫苗菌株,分别制备了2种菌体浓度的Mh和Pm单价灭活菌苗及3种菌量配比（1∶1、2∶1和3∶1）的Mh-Pm二联灭活苗,以小鼠为模型,皮下多点免疫（0.2 mL）,加强免疫2次,免疫剂量均为首免的一半。首免后第7天及其后每隔5 d,小鼠尾静脉采血分离血清,ELISA方法检测抗体效价,三免后第20天,分别以Mh422或PmCQ2进行腹腔攻毒测定免疫保护效果。结果显示,所有小鼠接种疫苗均无不良反应,二免后第10天抗体达较高水平,三免后抗体水平持续升高,第15天到达高峰,其后25 d维持高水平,后缓慢下降。Mh单菌苗的2种免疫剂量对Mh422株攻毒的免疫保护率均为0,而Pm单菌苗的2种免疫剂量对PmCQ2株攻毒的免疫保护率全为100%;Mh和Pm间无交叉免疫保护作用;3种菌量配比的Mh-Pm二联疫苗对Mh422株和PmCQ2株攻毒的各自免疫保护率分别为53%～71%和100%。该研究结果表明,所制备的Mh422单菌苗对同型攻毒无免疫保护作用,在诱导机体抗体产生方面,Mh和Pm间无相互抑制作用,PmCQ2株具有促进Mh422株灭活疫苗对Mh422的免疫保护作用,这为牛溶血性曼氏杆菌和牛多杀性巴氏杆菌二联疫苗的进一步研究提供了理论基础。     

猪流行性腹泻病毒全病毒灭活疫苗量效关系研究

本研究旨在探索猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)灭活疫苗不同抗原含量对仔猪感染的保护作用。测定不同浓缩倍数PEDV感染性病毒粒子数和病毒粒子总数,用不同浓缩倍数抗原分别制备灭活疫苗免疫小鼠,利用ELISA抗体检测方法、中和试验、ELISPOT方法测定体液免疫与细胞免疫产生情况,筛选合适抗原含量疫苗进行仔猪免疫并测定抗体,利用攻毒试验研究浓缩疫苗与保护之间的关系。结果显示,当抗原含量达8×10⁶ pfu·mL^-1以上时,灭活疫苗能有效刺激小鼠产生体液免疫及细胞免疫。以2 mL含8×10⁶ pfu·mL^-1抗原的灭活PEDV疫苗免疫仔猪可抵抗PEDV攻毒引起的腹泻及连续排毒,相较于总病毒粒子数,感染性病毒粒子数与抗体产生呈显著相关(r = 0.998 1),中和效价与仔猪保护呈显著相关性(r=0.974 7)。PEDV灭活疫苗可以提供良好的免疫保护,其中感染性病毒数量是提升抗体水平的关键因子,抗体滴度作为一项体液免疫的指标是判断免疫保护的重要参考。     

鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗对禽流感H9亚型流行株攻毒的保护作用

为了监测鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(LaSota株+M41株+SS/94株)对H9亚型禽流感病毒流行毒株的免疫保护效果,采用H9亚型禽流感病毒SS/94株及2009—2010年现地分离的3株H9亚型禽流感病毒对已免疫上述三联灭活苗的SPF鸡进行攻毒试验。结果显示,试验鸡以0.3 mL/只的剂量免疫三联灭活苗后21 d,其H9亚型禽流感病毒的HI抗体效价可达8～11log2,此抗体水平可抵抗2×106EID50的H9亚型禽流感病毒SS/94株、BLCN09株、WDZ09株、YT10株的攻击,攻毒保护率均达90%(9/10)以上。可见,以SS/94株作为禽流感疫苗抗原制备的三联灭活苗具有良好的免疫原性,能使免疫鸡抵抗2009—2010年期间现地分离的多株H9亚型禽流感病毒的攻击。     

Current perspectives on control of equine influenza

Influenza A viruses of the H3N8 subtype are a major cause of respiratory disease in horses. Subclinical infection with virus shedding can occur in vaccinated horses, particularly where there is a mismatch between the vaccine strains and the virus strains circulating in the field. Such infections contribute to the spread of the disease. Rapid diagnostic techniques are available for detection of virus antigen and can be used as an aid in control programmes. Improvements have been made to methods of standardising inactivated virus vaccines, and a direct relationship between vaccine potency measured by single radial diffusion and vaccine-induced antibody measured by single radial haemolysis has been demonstrated. Improved adjuvants and antigenic presentation systems extend the duration of immunity induced by inactivated virus vaccines, but high levels of antibody are required for protection against field infection. In addition to circulating antibody, infection with influenza virus stimulates mucosal and cellular immunity; unlike immunity to inactivated virus vaccines, infection-induced immunity is not dependent on the presence of circulating antibody to HA. Live attenuated or vectored equine influenza vaccines, which may better mimic the immunity generated by influenza infection than inactivated virus vaccines, are now available. Mathematical modelling based upon experimental and field data has been applied to examine issues relating to vaccine efficacy at the population level. A vaccine strain selection system has been implemented and a more global approach to the surveillance of equine influenza is being developed.     

猪繁殖与呼吸综合征免疫防制新途径的研究进展

猪繁殖与呼吸综合征(porcine reproductive and respiratory syndrome,PRRS)是一种主要表现为母猪繁殖障碍与仔猪呼吸道症状的传染病。近年来,猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)变异株不断出现,免疫逃避及持续性感染使得猪群发病率或复发率均相继增高,给养猪业带来了巨大的损失。目前所采用的胃肠道途径接种活疫苗或灭活疫苗的方法无法诱导对猪群的全面保护作用。为减少养猪业的经济损失,亟需研制新防制方法和新疫苗接种途径。作者主要从黏膜免疫的免疫部位、呼吸道保护性黏膜免疫反应诱导、黏膜免疫途径、佐剂的选择及病毒的免疫抑制反应等方面简要论述了有效防制PRRSV的黏膜免疫方法的研究进展,为进一步了解黏膜免疫抵御PRRSV突变株感染及黏膜疫苗研制等方面提供有用的信息。     

Mucosal vaccination with formalin-inactivated avian metapneumovirus subtype C does not protect turkeys following intranasal challenge

Studies were performed to determine if mucosal vaccination with inactivated avian metapneumovirus (aMPV) subtype C protected turkey poults from clinical disease and virus replication following mucosal challenge. Decreases in clinical disease were not observed in vaccinated groups, and the vaccine failed to inhibit virus replication in the tracheas of 96% of vaccinated birds. Histopathologically, enhancement of pulmonary lesions following virus challenge was associated with birds receiving the inactivated aMPV vaccine compared to unvaccinated birds. As determined by an enzyme-linked immunosorbent assay (ELISA), all virus-challenged groups increased serum immunoglobulin (Ig) G and IgA antibody production against the virus following challenge; however, the unvaccinated aMPV-challenged group displayed the highest increases in virus-neutralizing antibody. On the basis of these results it is concluded that intranasal vaccination with inactivated aMPV does not induce protective immunity, reduce virus shedding, or result in decreased histopathologic lesions.     

Protection against feline leukemia virus infection by use of an inactivated virus vaccine.

The protective immunity induced by 3 experimental FeLV vaccines were evaluated: Prototype inactivated FeLV vaccine developed from a molecularly cloned FeLV isolate (FeLV-FAIDS-61E-A); a mixture of immunodominant synthetic peptides corresponding to regions of the FeLV-Gardner-Arnstein-B (FeLV-GA-B) envelope proteins; and an adjuvant-disrupted but non-activated virus prepared from a non-cloned FeLV field isolate comprised of subgroup A and B viruses (FeLV-05821-AB). Included as controls were parallel groups of cats inoculated with adjuvants alone or with an established commercial FeLV vaccine. After each inoculation and after virulent virus challenge exposure, sera from all cats were assayed for ELISA-reactive antibody against purified FeLV, FeLV neutralizing (VN) antibody, and FeLV antigenemia/viremia--viral p27 antigen in serum and within circulating leukocytes. Immunity was challenged by oral/nasal exposure of vaccinated and control cats with FeLV-FAIDS-61E-A or FeLV-05821-AB, an infective, noncloned, tissue-origin, FeLV field isolate containing subgroup-A and -B viruses. Vaccine-induced immunity was assessed by comparing the postchallenge-exposure incidence of persistent viremia and the pre- and postchallenge exposure titers of VN and ELISA antibody in cats of the control and vaccine groups. The percentage of cats, that resisted development of persistent viremia after FeLV challenge exposure and the preventable fraction (PF) for the vaccine groups (which adjusts for the severity of the challenge and the degree of innate resistance in the controls) were as follows: adjuvant controls, 26%; FeLV-FAIDS-61E-A inactivated virus vaccine, 95% (PF = 93.2%); FeLV-GA-B peptide vaccine, 5% (-28.4%); FeLV-05821-AB noninactivated vaccine, 67% (55.4%); and commercial FeLV vaccine, 35% (12.2%). The prechallenge exposure mean VN antibody titer for each group was: less than 1:8 in the adjuvant controls; 1:43 in the FeLV-FAIDS-61E-A-vaccinated cats; less than 1:8 in the peptide-vaccinated cats; 1:38 in the noninactivated virus-vaccinated cats group; and 1:12 in the cats vaccinated with the commercial vaccine. Thus, induction of VN antibody in the vaccinated cats, although modest, appeared to be correlated with induction of protective immunity as defined by resistance to FeLV challenge exposure. Results of these studies indicate that inoculation of cats with an experimental inactivated virus vaccine prepared from a molecularly cloned FeLV isolate was most effective in stimulating protective immunity against heterologous and homologous FeLV challenge exposure.     

促益素雏鸡IBD疫苗免疫后局部体液的免疫变化

本研究分别将两种促益素“禽福”和“亿妙灵”与鸡传染性法氏囊病中等毒力活疫苗配合应用,通过间接ELISA法检测免疫雏鸡泪液、气管液,胆汁、肠液的IgG、IgM、IgA含量的动态变化。结果表明：应用促益素雏鸡IBD疫苗免疫后局部体液的上述三种免疫球蛋白含量较IBD疫苗单独免疫雏鸡有不同程度地升高：IBD强毒攻击后,疫苗单独免疫雏鸡局部体液的上述指标明显低于应用促益素的疫苗免疫雏鸡。表明使用促益素后,可提高疫苗增强机体局部免疫的功能,有效抵抗强毒攻击,使疫苗的免疫保护率显著提高：在两种促益素中,“禽福”的局部体液免疫增强效果强于“亿妙灵”。     

Evaluation of a 3A-truncated foot-and-mouth disease virus in pigs for its potential as a marker vaccine

Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of cloven-hoofed animals in the world. The disease can be effectively controlled by vaccination of susceptible animals with the conventional inactivated vaccine. However, one major concern of the inactivated FMD virus (FMDV) vaccine is that it does not allow serological discrimination between infected and vaccinated animals, and therefore interferes with serologic surveillance and the epidemiology of disease. A marker vaccine has proven to be of great value in disease eradication and control programs. In this study, we constructed a marker FMDV containing a deletion of residues 93 to 143 in the nonstructural protein 3A using a recently developed FMDV infectious cDNA clone. The marker virus, r-HN/3A_93–143, had similar growth kinetics as the wild type virus in culture cell and caused a symptomatic infection in pigs. Pigs immunized with chemically inactivated marker vaccine were fully protected from the wild type virus challenge, and the potency of this marker vaccine was 10 PD₅₀ (50% pig protective dose) per dose, indicating it could be an efficacious vaccine against FMDV. In addition, we developed a blocking ELISA targeted to the deleted epitope that could clearly differentiate animals infected with the marker virus from those infected with the wild type virus. These results indicate that a marker FMDV vaccine can be potentially developed by deleting an immunodominant epitope in NSP 3A.     

鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活 疫苗(La Sota株+M41株+SS/94株)免疫程序研究

设计不同的免疫程序,用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)免疫黄羽肉鸡,通过对ND、IB、H9抗体滴度监测,探讨ND、IB、H9抗体消长规律及鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)的免疫程序。试验结果表明,用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)免疫黄羽肉鸡后,其诱导产生的ND、IB、H9抗体的消长规律基本同步;仅于10日龄免疫一次三联灭活苗,其抗体水平较低,于20、40日龄二免、三免或10日龄先用鸡新城疫病毒(La Sota株)、禽流感病毒(H9亚型,SS/94株)二联灭活疫苗作基础免疫,20或40日龄再用三联灭活苗作加强免疫,则上述3种抗体均快速上升,且维持时间长。根据试验结果,建议按照正常免疫程序作基础免疫的健康肉鸡,饲养期较短的可于20日龄左右用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)作加强免疫,0.3 mL/只;饲养期较长的则于40日龄左右用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)作加强免疫,0.5 mL/只。     

猪繁殖与呼吸综合征灭活疫苗的研制

本研究选用国内分离的猪繁殖与呼吸综合征病毒ＣＨ－１ａ株作为疫苗用毒株。利用转瓶细胞培养技术,在Ｍａｒｃ－１４５细胞上大量增殖了较高滴度的猪繁殖与呼吸综合征病毒。该病毒经甲醛灭活后用等量的油佐剂乳化制成安全性好、保护率高的猪繁殖与呼吸综合征油佐剂灭海灭凤苗。经实验试验和区域性跟踪试验证明,该设苗具有免疫保护工,保护率高、安全稳定等优点。