抗猪瘟免疫球蛋白在猪初乳中变化规律研究

以猪瘟弱毒疫苗免疫空怀母猪,待母猪分娩后,分别收集10d内的乳汁,以建立的间接ELISA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,7d后抗体水平与常乳基本一致(>0.05)。对照组母猪仅在分娩当天检测到较低的抗体水平。     

猪乳中猪瘟IgG、IgM抗体与猪瘟母源抗体变化规律

选取母猪20头,分为2组,10头／组。免疫组母猪分别于配种前25d、10d进行2次猪瘟疫苗的免疫接种,4头份／次,肌肉注射。对照组接种疫苗稀释液,1mL／头。待母猪分娩后3h采集乳样,并分别于产后2d、3d、4d、5d、6d、7d、8d、9d、10d、15d、20d、25d、30d采集乳样。用间接ELISA方法,测定猪瘟IgG、IgM抗体OD值,并绘制动态变化曲线,观察抗体动态变化规律。待上述免疫组母猪产仔后,各组随机选取10头仔猪,仔猪分别于产后0（未吃初乳）、5、10、15、25、35、40、50、60、70、80、90日龄前腔静脉采血,分离血清,用间接ELISA法检测其母源猪瘟抗体动态变化规律。结果表明：免疫组母猪乳清中IgG、IgM抗体效价的最高值均出现在分娩后1d,随后抗体效价迅速降低,一般泌乳7d以后,免疫组母猪乳中各种抗体效价与常乳基本一致。初生仔猪的母源抗体在5～10日龄达到高峰,10日龄后开始下降,至25日龄保护率已达不到100％,抗体水平接近保护线。     

猪瘟免疫和抗体消长规律的跟踪试验

猪瘟是我国养猪业的主要传染病之一 ,具有传染性强、流行广、发病率和死亡率高等特点 ,是我国目前进行强制免疫的主要传染病之一。我国生产的猪瘟弱毒疫苗享誉国内外 ,为世界上许多国家消灭和控制猪瘟起了重要作用。近年来 ,我国某些地区出现了以温和型为主的猪瘟 ,本市在抗体监测过程中同样发现猪瘟抗体水平低下现象。本文通过抗体跟踪监测和不同剂量比较试验等 ,系统地反映猪瘟免疫注射后抗体的消长规律和免疫效果。1　材料和方法1 .1　材料猪瘟阳性血清 (中国农科院兰州兽医研究所 ,批号 :2 0 0 10 80 9、2 0 0 2 0 4 30 )。猪瘟阴性血清…     

猪瘟疫苗的研究进展

猪瘟疫病对我国养猪业的危害很大,防控猪瘟主要依靠疫苗免疫,目前市场猪瘟疫苗很多,一般分为弱毒活疫苗和新型猪瘟疫苗,研究者们主要关注的为新型猪瘟疫苗。文章对现市场上的猪瘟疫苗及正在研究的猪瘟疫苗进行了综述,为猪瘟疫苗的使用和进一步研究提供借鉴与参考。     

卡介苗不同用药途径对猪瘟弱毒疫苗调节作用的研究

将30头20日龄法国大白仔猪，随机分为山。A-2、A-3、B-1、B-2、C六个组．每组5头．以研究活或灭话卡介苗（BC）对猪瘟弱难疫苗的调节作用，结果表明：皮内、皮下注射活或灭活BCG均可显著地提高猪瘟弱毒酶联单克隆抗体水平．     

猪母乳抗E2抗体水平的调查研究

本实验对采至广西某猪场的270份母乳奶样,应用间接ELISA和间接血凝试验,分别对其进行了抗猪瘟E2抗体水平和猪瘟全抗体水平的检测.检测结果显示,在270份样品中,间接ELISA检测呈阳性的(即含有抗E2抗体的)占总样品的94.45%,而间接血凝试验检测出这些样品的效价在1:16以上(具有保护力的)高达97.78%.从实验结果可以看出,该猪场母猪在哺育阶段的母乳含有抗E2抗体,仔猪能从母乳获得抗体,从而获得对猪瘟的免疫力.从两个试验的检测结果对照可知,两个数据差异不显著,说明用间接ELISA检测抗E2抗体、用间接血凝试验检测猪瘟抗体来监测猪瘟抗体免疫水平是可行的.本实验进一步论证了E2抗体就是猪瘟的保护性抗体,同时为监测猪瘟开创了一种新的可行方法,这对监测猪瘟真正的免疫水平,帮助猪场制定有效的免疫程序具有很现实的重要的意义.     

皮下注射鸭瘟弱毒苗诱导鸭局部黏膜和系统免疫中IgA、IgM和IgG的发生规律

为探讨鸭瘟弱毒苗诱导鸭局部黏膜和系统免疫中抗体发生的规律,将鸭瘟病毒Cha株弱毒苗皮下注射免疫20日龄樱桃谷鸭后60 d内定时随机剖杀5只鸭,采集血清、胆汁、气管和消化道(食道、十二指肠、空肠、回肠、盲肠和直肠)分泌液,应用间接ELISA检测抗DPV的IgA、IgM和IgG滴度(以Log10表示).结果表明:①血清:抗体滴度由高到低为IgG、IgM和IgA,相应的检测到的时间段分别为免疫后6～60,3～15,12～36 d.②胆汁:抗体滴度由高到低为IgA、IgG和IgM,相应的检测到的时间段分别为免疫后9～21,15～27,3～12 d.③分泌液:气管和消化道分泌液中抗体滴度由高到低均为IgA、IgM和IgG,其中IgA抗体滴度由高到低为十二指肠、食道、气管、空肠、盲肠、回肠和直肠,相应的检测到IgA的时间段分别为免疫后3～60,9～60,3～60,9～60,12～60,12～27,6～36d;IgM由高到低为气管、食道、十二指肠、空肠、盲肠、直肠和回肠,相应的检测到IgM的时间段分别为免疫后3～12,6～15,3～12,6～12,9～12,6～9,6～9 d;IgG由高到低为食道、十二指肠、气管、空肠、盲肠、直肠和回肠,相应的检测到IgG的时间段分别为免疫后9～36,12～27,6～36,9～36,12～36,9～21,15～21 d.综上,鸭瘟弱毒疫苗皮下免疫鸭后,IgM和IgG分别是系统免疫中体液免疫的先锋抗体和主要抗体;IgA是气管、消化道和胆汁中的主要抗体.     

Protection of gruntlings against classical swine fever virus-infection after oral vaccination of sows with C-strain vaccine

The objective of this study was to investigate the maternal protection of gruntlings derived from wild sows vaccinated orally against classical swine fever (CSF) using C-strain vaccine. Three vaccinated sows and one unvaccinated control sow were included. Challenge infection of the progeny was carried out either intranasally or by contact at the beginning of the third month of life (61-65 days post-natum). Whereas, two of three litters had maternal antibodies, the progeny of one vaccinated sow was seronegative at challenge. The progeny of the control sow, which was challenged by contact infection, developed moderate clinical signs except for one animal which became ill and died. Two gruntlings derived from the vaccinated sows also died of CSF, although one of them had a relatively high maternal antibody titre (128 ND(50)). The transient infection and partial virus shedding observed in a small number of gruntlings with maternal antibodies and the fact that one animal with maternal antibodies became ill and died confirm the incomplete maternal protection at this age. The reason for this incomplete protection is discussed. As none of the surviving gruntlings could be shown to carry CSFV or viral RNA at the end of the experiment (36 or 70 d.p.i.), it may be concluded that these animals do not represent a potential CSFV reservoir.     

IgG, IgM and IgA in the serum of cattle naturally infected with Mycobacterium paratuberculosis

Serum IgG, IgM and IgA antibody response in 20 cattle naturally infected with Mycobacterium paratuberculosis and in 15 non-infected cattle were measured by enzyme-linked immunosorbent assay. A strong IgG response was detected in 16 (80%) of the infected animals. Diagnostic levels of IgM were detectable in all of the infected animals as well as in 8 (53%) of the non-infected animals. Animals with paratuberculosis had a very weak specific serum IgA response and this appears to be of little value in detection of infection in these animals.     

猪瘟兔化弱毒疫苗株基因组的遗传变异分析

参照已发表的猪瘟病毒基因组序列设计了9对引物，用RT-PCR从猪瘟兔化弱毒疫苗株细胞培养物中扩增得到了覆盖猪瘟病毒基因组全长的9个cDNA片段，将所得cDNA片段分别克隆至pMD18-T载体中，经测序和拼接后，获得了猪瘟兔化弱毒疫苗株基因组全序列。序列分析表明，猪瘟兔化弱毒疫苗株基因组全长12310个碱基，其5’非编码区（5＇-NCR）和3＇-NCR分别由373和239个碱基组成，在3’末端有富含T的碱基插入，其间为1个大的开放阅读框架，编码3898个氨基酸残基的多聚蛋白，与国内外已发表的另外7个猪瘟兔化弱毒疫苗株基因组全序列相比，核苷酸同源性为98．7％～99．9％，氨基酸同源性为98．6％～99．9％。基因组全序列比较显示，猪瘟兔化弱毒疫苗株基因组在遗传上相当稳定。     

Serum concentrations of IgG, IgA, and IgM in retired racing Greyhound dogs

Background: Greyhound dogs have significant physiologic, hematologic, and biochemical differences when compared with other breeds, including significantly lower serum globulin concentration owing to decreases in the α‐ and β‐globulin fractions. The specific proteins that account for differences in globulin concentrations are not known, but IgA and IgM, both β‐globulins, are potential candidates. Objectives: The aims of this study were to measure serum IgG, IgA, and IgM in clinically healthy retired racing Greyhounds and compare the results with those of age‐ and sex‐matched non‐Greyhound dogs. Methods: Study animals included 25 Greyhound and 20 non‐Greyhound dogs. Total protein, albumin, and total globulin concentrations were determined. IgG, IgA, and IgM concentrations were measured using a commercially available radial immunodiffusion kit. The Student t‐test assuming equal variances was used to compare concentrations of immunoglobulins between groups. Results: Serum concentrations of IgA and IgM in Greyhounds (IgA=49±20 mg/dL; IgM=132±47 mg/dL) were significantly lower than concentrations in non‐Greyound dogs (IgA=70±39 mg/dL; Ig M=212±78 mg/dL). Concentrations of IgG did not differ between groups. Conclusions: Mean serum IgA and IgM concentrations in Greyhounds were lower than those in non‐Greyhound dogs. This may contribute to low serum concentrations of β‐globulins in Greyhounds. Specific reference intervals are recommended for Greyhounds to avoid possible misdiagnosis of IgA or IgM deficiency.     

口服鸭瘟弱毒苗诱导鸭局部黏膜和系统免疫中IgA、IgM和IgG发生规律研究

为探讨鸭瘟弱毒苗诱导鸭局部黏膜和系统免疫中抗体发生的规律,本研究将鸭瘟病毒Cha株弱毒苗经口免疫20日龄樱桃谷鸭,60 d内定时随机剖杀5只鸭,采集血清、胆汁、气管和消化道(食道、十二指肠、空肠、回肠、盲肠和直肠)分泌液,应用间接ELISA检测抗DPV的IgAI、gM和IgG滴度(以Log10表示)。结果表明:①血清中检测到IgA、IgM和IgG的时间段分别为9-36、3-15和9-60天,滴度由高到低为IgG、IgM和IgA;②胆汁中检测到IgAI、gM和IgG的时间段分别为6-15、9-12和12-36天,滴度由高到低为IgA、IgG和IgM;③气管中检测到IgA、IgM和IgG的时间段分别为6-60、3-12和9-27天,滴度由高到低为IgAI、gM和IgG;④食道中检测到IgAI、gM和IgG的时间段分别为3-60、3-27和9-60天,滴度由高到低为IgA、IgM和IgG;⑤各肠段抗体滴度由高到低及相应检测到的时间段:十二指肠中为IgA(3-60天)I、gM(3-15天)和IgG(9-27天);空肠中为IgA(6-60天)I、gM(6-12天)和IgG(9-36天);回肠中为IgA(9-60天)、IgM(6-9天)和IgG(15-21天);盲肠中为IgA(6-60天)I、gG(12-36天)和IgM(6天);直肠中为IgA(3-60天)I、gG(12-21天)和IgM(6天)。结果显示,鸭瘟弱毒疫苗经口免疫鸭后,IgM和IgG分别是系统免疫中体液免疫的先锋抗体和主要抗体;IgA、IgG和IgM在胆汁中存留时间短;IgA是气管和消化道中的主要抗体。     

富锗种蛋对其初生雏鸡血清IgG,IgM水平的影响

通过给父母代种鸡饲喂四个不同梯度剂量的Ge-132,以获取富锗种蛋,分组孵化后,各取初生雏鸡100只定为试验Ⅰ、Ⅱ、Ⅲ、Ⅳ组;另取普通种蛋孵出的初生雏鸡100只,设为第Ⅴ组,以作对照。各组再分成两个小组,每小组50只,其中Ⅰ-1、Ⅱ-1至Ⅴ-1小组按常规进行免疫处理;而Ⅰ-2、Ⅱ-2至Ⅴ-2小组则不进行免疫。于1、7、14、21、28日龄时分别采血、检测血清IgG、IgM含量,结果显示:1日龄雏鸡血清IgG、IgM含量Ⅰ～Ⅳ显高于第Ⅴ组,差异极显著(P<0.05);1日龄后,Ⅰ～Ⅳ组抗体水平逐渐下降,至7日龄时,降至低谷并接近对照组;其后,至28日龄间各组雏鸡血清IgG,IgM含量又呈上升趋势,而Ⅰ～Ⅳ组始终高于第Ⅴ组、但差异并不明显(P>0.05);各免疫处理小组与非免疫小组比较,二者间未见明显的、规律性差异。实验结果首次证实:富锗种蛋孵出的雏鸡,其血清IgG、IgM水平明显高于普通雏鸡,尤于1周龄阶段内更为显著。表明Ge-132能明显地提高初生雏鸡的抗体水平。     

猪瘟活疫苗效力检验替代方法的研究

目前,我国猪瘟活疫苗效力检验最常用的方法是测定猪瘟疫苗中病毒的兔体感染量(RID),但是用该方法易受到检验兔品种、个体和饲养环境的影响。为了探索一种不依赖动物的疫苗效力检验新方法,本研究将待检猪瘟活疫苗系列稀释后分别接种易感细胞,使用抗原捕获ELISA、RT-nest PCR和RT-PCR三种方法检测不同稀释度疫苗中活病毒在感染细胞后增殖的子代病毒,计算疫苗病毒的最高细胞感染剂量,建立了猪瘟疫苗病毒的细胞感染量(CID)的效力检验方法。结果显示:疫苗中活病毒粒子越多,CID就越高;对不同类型猪瘟疫苗的检测结果表明,该方法适用于传代细胞源和细胞源猪瘟活疫苗的效力效检。使用CID和RID两种检测方法同时对12批猪睾丸传代细胞源(传代细胞源)和3批牛睾丸原代细胞源(细胞源)猪瘟活疫苗进行了比对效检,结果表明,用CID测定方法检验,12批猪瘟活疫苗(传代细胞源)每头份均含105CID,3批猪瘟活疫苗(细胞源)每头份含量均为104CID;用兔子感染体温测定法,12批猪瘟活疫苗(传代细胞源)每头份含量在2.6×104~3.0×104RID之间,3批猪瘟活疫苗(细胞源)每头份含7.0×103~8.0×103RID。试验证明:本实验室建立的CID检测结果与现有质量标准RID的结果存在良好的相关性,能够较好反映疫苗中活病毒的含量,有望成为RID的替代方法。