3种免疫程序对不同规模猪场仔猪猪瘟抗体水平影响的观察

猪瘟是由猪瘟病毒引起的猪一种急性或慢性、热性和高度接触性传染病,是目前危害我国养猪业最严重的传染病,尽管我国成功研制了最好的猪瘟疫苗,并且在猪瘟的防治方面发挥了积极作用,但目前猪瘟疫苗免疫效果却往往不尽如人意,其中免疫程序不合理是主要原因.因此,我们应用3种免疫程序对不同规模猪场仔猪的免疫效果进行了监测,旨为不同规模猪场提供比较科学、合理的免疫程序,以提高猪瘟的防控水平. 1 试验材料 1.1 检测试剂盒 猪瘟病毒ELISA抗体检测试剂盒,购自武汉科前动物生物制品有限责任公司.     

吮吸初乳时间对新生仔猪猪瘟疫苗免疫的影响效果

出生后立即给仔猪注射2头份的猪瘟疫苗，间隔一定时间后再让其吮吸初乳，其中Ⅰ～Ⅳ组间隔时间为120,90，60，30min，V组为对照组不注射疫苗。在20日龄时用猪瘟Dot-ELISA诊断试剂盒对这些仔猪血清中的猪瘟抗体滴度进行测定。结果：在接种疫苗后60—120min之间吮吸初乳的免疫效果最好。     

某规模化猪场猪瘟抗体水平的监测

为了对某猪场猪瘟疫苗免疫效果做出评估并进一步优化免疫程序,应用猪瘟疫苗抗体阻断ELSIA检测试剂盒对该场不同日龄仔猪及育肥猪的猪瘟疫苗抗体水平进行检测,对抗体合格率、平均阻断率和变异系数统计分析。检测数据显示,该猪场猪瘟免疫空白期过长,疫苗免疫接种后抗体合格率较低,增加了感染猪瘟病毒的风险,应及时进行猪瘟免疫程序的优化。猪瘟疫苗抗体水平监测对猪瘟防控具有重要意义,可为规模化猪场开展猪瘟疫苗抗体检测,制定合理的免疫措施提供参考。     

O型口蹄疫疫苗与猪瘟疫苗同步免疫后免疫抗体群体合格率监测

猪口蹄疫和猪瘟均是严重危害养猪业生产的重大动物疫病,目前我国对猪口蹄疫和猪瘟最有效的预防和控制措施就是进行免疫接种。本研究采用O型口蹄疫液相阻断ELISA试验和猪瘟正向间接血凝试验方法对猪O型口蹄疫疫苗、猪瘟疫苗单独免疫及猪O型口蹄疫疫苗与猪瘟疫苗同步免疫后O型口蹄疫免疫抗体和猪瘟免疫抗体群体合格率分别进行监测,以期摸索出沈阳地区猪口蹄疫和猪瘟疫苗同步分点免疫后抗体群体保护率     

猪瘟疫苗在猪场的免疫评估

在生猪养殖过程中,猪瘟属于常见疾病,具有较强的传染性,所以疫苗免疫成为防控猪瘟的关键措施。制定科学合理的免疫程序对猪瘟防控尤为重要。但实际生产中,猪场免疫效果,并不是太乐观,需要不断优化免疫程序。利用先进的试剂盒技术来及时检测猪瘟疫苗的免疫效果,以便及时调整疫苗使用种类,不断优化免疫程序,减少经济损失。     

猪强制性免疫中不同免疫程序的免疫效果调查分析

为探讨口蹄疫O型灭活苗、猪瘟活疫苗、高致病性猪蓝耳病活疫苗不同免疫程序的免疫效果。我们选择了6个规模养猪场,对其采用的2种免疫程序的免疫效果进行调查分析。调查分析结果表明,高致病性猪蓝耳病活疫苗的免疫对猪瘟疫苗和口蹄疫疫苗的免疫具有一定免疫干扰或抑制作用;免疫程序中,高致病性猪蓝耳病活疫苗先于猪瘟疫苗和口蹄疫疫苗免疫,其猪群中猪瘟与口蹄疫和高致病性猪蓝耳病的疫苗免疫效果均好;猪瘟疫苗和口蹄疫疫苗同时分点肌内注射,猪群中猪瘟与口蹄疫免疫抗体平均合格率可达到70%以上。建议在实际免疫工作中对生猪进行免疫时,高致病性猪蓝耳病活疫苗要先于猪瘟疫苗和口蹄疫疫苗免疫,且猪瘟疫苗和口蹄疫疫苗可同时分点肌内注射。     

通过抗体检测建立合理的猪瘟、伪狂犬病免疫程序

随着我国养猪业的飞速发展 ,防病治病技术也有了很大提高 ,但是 ,仍有不少猪场存在免疫程序不合理现象。金华种猪场以抗体测定试验为依据 ,对原来的猪瘟、伪狂犬病的免疫程序作相应的调整。试验情况总结如下。1　材料和方法1 1　试验材料血清样为金华种猪场猪群中抽取的血样 ;猪瘟正向血凝诊断液 (中国农科院兰州兽医研究所生产 ) ;猪伪狂犬病乳胶凝集诊断试剂盒 (华中农业大学牧医学院生产 ) ;猪瘟疫苗 (江西民星集团公司江西生物药厂的猪瘟兔化弱毒苗 ) ;伪狂犬病基因缺损疫苗 (荷兰英特威公司生产 )1 2　试验方法自 1998年 6月开始 ,每隔…     

不同猪瘟活疫苗免疫效果比较分析

<正>猪瘟是由黄病毒科瘟病毒属的猪瘟病毒引起的一种急性、热性、高度接触性传染病。近年来,猪瘟已被消灭的一些欧洲国家又报道有猪瘟发生,我国猪瘟的发病率亦呈上升趋势,严重威胁着我国养猪业的发展[1]。当前,预防猪瘟最好的方法就是免疫猪瘟疫苗。为了探索最佳的猪瘟疫苗免疫程序和免疫质量,指导猪瘟疫苗免疫预防工作,更好地对重大动物疫病进行防控,有必要对国家采购的猪瘟疫苗进行免疫程序和免疫效果的评估。     

福建省猪瘟防制应强化的几个方面

通过对猪瘟的监测和流行病学调查表明,猪瘟仍是目前造成福建省猪死亡的主要原因。由单一发生向混合感染趋势发展,猪瘟在有限区域散发性流行,以非典型症状猪瘟为主,部分猪场由于猪免疫抑制病等因素造成种猪场以母猪带毒隐性感染引起母猪繁殖障碍和仔猪死亡,多数猪场采用大剂量疫苗注射。福建省在防制猪瘟方面应筛选猪瘟疫苗,制定和推广科学免疫程序;建立全省猪瘟监测网络和报警系统,加强对种猪场、规模猪场和散养猪的免疫、监测,尤其对种猪场开展猪瘟免疫抗体与野毒感染抗体,疫苗毒与野毒抗原的鉴别诊断,坚决淘汰免疫缺陷和野毒感染的种猪,在种猪场推行猪瘟净化技术;加强对生猪市场交易、调运的管理。     

猪瘟与伪狂犬疫苗联合使用免疫效果评估试验研究

由于目前我国猪疫病发生率较高,疫苗免疫程序相对复杂,为了简化免疫程序减少疫苗接种带来的过度应激,本试验主要针对商品猪猪瘟和伪狂犬疫苗在接种日龄上和疫苗生产工艺相近的特点,对猪瘟和伪狂犬疫苗在商品猪上联合使用,以此掌握猪瘟和伪狂犬疫苗联合使用后对猪群的保护力和是否对猪群造成不良的应激反应。试验结果表明猪瘟与伪狂犬疫苗联合免疫后,猪瘟二免后阻断率在70%以上,通过两次联合免疫后保护期能达到160日龄以上,伪狂犬g B均为阳性,伪狂犬g E均为阴性,通过两次联合免疫后保护期能达到160日龄以上。说明猪瘟和伪狂犬疫苗联合免疫可以给猪群提供良好的保护,与单独免疫比较基本无差别。本试验通过测定猪瘟疫苗和伪狂犬疫苗联合免疫后抗体消长规律和应激反应,为今后猪瘟和伪狂犬疫苗在临床广泛应用提供基础数据。     

囊虫病免疫诊断检测方法的研究进展

由于科学技术的发展,囊虫病免疫诊断检测方法有了很大的进展,主要有1.免疫学检测方法,包括酶联免疫吸附法(ELISA)、斑点酶联免疫吸附法(Dot-ELISA)、生物素-亲和素酶联免疫吸附法(BAS-ELISA)、单克隆抗体酶联免疫吸附试验(McAb-ELISA)、酶联免疫印渍技术(ELIB);2.基因检测技术,包括DIG标记DNA探针、细胞因子基因检测方法、基因重组技术;3.免疫试剂盒的应用,包括循环抗原酶联免疫试剂盒、猪囊虫短程抗体IgG4快速ELISA检测试剂盒等。     

Does cortisol bias cytokine production in cultured porcine splenocytes to a Th2 phenotype?

Glucocorticoids are reported to bias the production of cytokines from a type 1 to a type 2 phenotype. However, this dogma has been advanced largely from studies utilizing potent glucocorticoid analogs, particularly dexamethasone (DEX). Although studies utilizing DEX certainly have clinical and pharmacological relevance, DEX is probably not the best glucocorticoid for studies designed to evaluate the interaction and regulation of endogenous corticosteroids with immune cells in vivo in the domestic pig. Functional measures of immune suppression suggest that the pig is relatively resistant to DEX. Furthermore, type II corticosteroid receptors exclusively bind DEX with high affinity, whereas type I receptors, the so-called mineralocorticoid receptors, have a higher affinity for cortisol. In addition, DEX is not bound by serum binding proteins as are endogenous corticosteroids.These issues prompted us to revisit glucocorticoid regulation of type 1 and type 2 cytokines in cultured pig splenocytes and to test the broad hypothesis that cortisol biases cytokine production in favor of a Th2 response. We evaluated interferon gamma (IFNgamma) (also interleukin 2 (IL-2) in one experiment) and interleukin 10 (IL-10) as representative Th1 and Th2 cytokines, respectively. Furthermore, we evaluated macrophage migration inhibitory factor (MIF) because it is reported to be an essential factor in T cell activation; it is also upregulated by glucocorticoids and reported to be a product of Th2 lymphocytes. In general, both IFNgamma and IL-10 were sensitive to cortisol inhibition early in culture. However, IFNgamma ultimately escaped cortisol inhibition, whereas IL-10 continued to be substantially suppressed by high physiological concentrations of cortisol. Similarly, MIF mRNA could be suppressed by cortisol, but only when cortisol was added to cultures after ConA (concanavalin A) stimulation of splenocytes. So, taken together, our studies do not support the hypothesis that cortisol favors a Th2 cytokine profile in cultured pig splenocytes.     

Natural infection of pigs with bovine viral diarrhea virus and its differential diagnosis from hog cholera.

Natural infection of pigs with bovine viral diarrhea virus (BVDV) through contact with infected cattle has caused problems in diagnosing hog cholera (HC). Low cross-reacting serum antibody titers against HC caused by BVDV infection were found in clinically normal pigs as well as those suspected of having HC. Bovine viral diarrhea virus was isolated from specimen tissues and initially identified as HC virus (HCV), using the fluorescent antibody cell culture technique. Additional cell cultures, as well as pig and calf trials, were necessary to identify it as BVDV. The isolate caused clinical signs of illness in the calves, whereas the pigs remained healthy. Bovine viral diarrhea virus may be detected in tissue sections or isolated in cell cultures and confirmed as HCV, using the HC fluorescent antibody conjugate. Laboratories performing the neutralization test for HC should use discretion when interpreting HC titers unless BVD titers are determined on the same serums.     

Recent applications of the Dot-ELISA in immunoparasitology

The dot enzyme-linked immunosorbent assay (Dot-ELISA) is a highly versatile solid-phase immunoassay for antibody or antigen detection. The assay uses minute amounts of reagent dotted onto solid surfaces such as nitrocellulose and other paper membranes which avidly bind proteins. After incubation with antigen-specific antibody and enzyme-conjugated anti-antibody, the addition of a precipitable, chromogenic substrate causes the formation of a colored dot on the solid phase which is visually read. The Dot-ELISA has been used extensively in the detection of human and veterinary protozoan and metazoan parasitic diseases, including amebiasis, babesiosis, fascioliasis, cutaneous and visceral leishmaniasis, cysticercosis, echinococcosis, malaria, schistosomiasis, toxocariasis, toxoplasmosis, trichinosis, trypanosomiasis and even ixodid tick infestation. The technique is rapid, easy to perform and interpret, reagent conservative, cost effective and field portable. In addition, the Dot-ELISA may be configured to detect antibodies or parasite antigen in either microtiter plates for large-batch testing or with dipsticks for small numbers of determinations. A slight modification of the Dot-ELISA procedure allows the determination of infection rates of vectors such as ticks and sandflies with parasites.     

Detection of Campylobacter antibodies in sheep sera by a Dot-ELISA using acid extracts from c. fetus ssp. fetus and c. jejuni strains and comparison with a complement fixation test

In this study, a dot-enzyme-linked immunosorbent assay (Dot-ELISA) was evaluated in comparison with a complement fixation test (CFT) for the detection of Campylobacter antibodies in sheep sera. Acid glycine extracts (AGE) of both Campylobacter fetus ssp. fetus and Campylobacter jejuni strains that had been isolated from the gall-bladder of slaughtered sheep was used as antigen in both tests. A total of 153 sheep sera from aborted (74) and slaughtered (79) sheep were examined by both Dot-ELISA and CFT. Twenty-two sera showed anti-complementary activity were not suitable for CFT. Of the 22 sera showing anti-complementary activity, two sera were found to be positive in Dot-ELISA. Eighty-eight (67.2%) of the remaining 131 sera were negative by both Dot-ELISA and CFT using AGE of both Campylobacter strains whereas 43 sera (32.8%) gave different reaction patterns in Dot-ELISA and CFT with the extracts of both Campylobacter strains. Twelve sera were positive by both tests using AGE of C. fetus ssp. fetus but CFT failed to detect antibodies in nine of these sera when AGE of C. jejuni was used. Twelve sera were positive by both tests only when AGE of C. fetus ssp. fetus was used. Eleven sera were positive only by CFT. Seven of these reacted only with the AGE of C. fetus ssp. fetus and four sera were positive by using AGE of both Campylobacter strains. The remaining eight sera were found to be positive only by dot-immunobinding assay either with the AGE of both Campylobacter strains or with the AGE of one of the Campylobacter strains. It is concluded that Dot-ELISA using AGE from C. fetus ssp. fetus could be employed for the detection of Campylobacter antibodies in sheep sera and the additional use of AGE from C. jejuni as antigen appeared not to be profitable for this purpose.     

我国规模化猪场对猪伪狂犬病的控制

猪伪狂犬病是由猪伪狂犬病病毒引起的一种急性传染病。规模化猪场应根据野毒感染的严重程度采取切合实际的控制策略,疫苗免疫是重要的控制手段。常规灭活苗、常规弱毒苗、基因缺失疫苗都有不同的特点和使用范围,经过分析国内规模化猪场使用基因缺失疫苗是最好的选择。笔者在免疫程序及方式方面对有争议的问题如首免日龄、超前免疫、免疫间隔时间、滴鼻免疫等进行了分析;强调了免疫监测的重要性。提出了规模化猪场净化猪伪狂犬病的措施及可行性,总结了国内某些场成功的经验,为猪场控制该病提供实践参考。     

FMD、HC和PRRS疫苗不同组合接种免疫效果的此较

为了研究猪O型口蹄疫(FMD)、猪瘟(HC)、高致病性猪蓝耳病(PRRS)疫苗不同组合的免疫效果,在黔东南州3个养殖场各选择35日龄仔猪120头.每个场分成6个实验组,每组20头,在免疫后的21、28、35、42、49、56、63 d采血检测抗体,FMD、HC用正向间接血凝试验检测,PRRS用间接EIJSA检测.结果表...     

6个猪品种CD53基因中结构变异SV184分布频率的研究

基于香猪全基因组重测序数据,在白细胞表面抗原CD53基因中筛选到一个结构变异（structure variant,SV）184,为了探索不同猪品种间此结构变异是否存在多态性变化,本试验选择香猪、大白猪、糯谷猪、柯乐猪、江口萝卜猪和荣昌猪作为试验动物,采用PCR方法对猪群中SV184的分布频率进行比较研究,应用RT-PCR技术,分析CD53基因的原初转录本序列结构,研究SV184对CD53基因的转录是否有直接影响。结果显示,6个猪品种中SV184的分布频率存在明显差异,香猪以VN基因型为主,其他5个猪品种是NN基因型占优势;计算两种等位基因的频率,香猪以缺失的V等位基因为主,其他5个猪品种以正常的N等位基因为主。选择3种基因型个体的血液样本,逆转录后分析CD53基因的原初转录本的序列,从中检测到缺失型和不缺失的两种转录本形式,提示SV184对CD53基因的转录无直接影响。SV184可作为分子标记用于区分香猪和其他猪品种。     

不同抗原对猪瘟病毒抗体ELISA检测效果的影响

为研究不同抗原对猪瘟病毒抗体ELISA检测效果的影响,通过原核表达获取猪瘟病毒重组E2蛋白、E0蛋白、C蛋白和NS5B蛋白,大小分别约为35 kD、42 kD、16 kD和80 kD。经Ni-NTA亲和层析柱纯化并利用BandScan软件进行计算,纯度均在90%以上,满足ELISA检测包被用原料纯度的要求。将上述4种蛋白作为包被抗原进行ELISA,以10份企业阳性质控品血清和10份企业阴性质控品血清为检验指标比较不同抗原对猪瘟病毒抗体ELISA检测效果的影响。结果以E2蛋白为原料的包被板检测质控血清时,灵敏度和特异性均达到80%以上,能够满足猪瘟病毒抗体ELISA检测的要求,故选择E2蛋白作为包被抗原并进行相关检测试剂的研制。用研制的试剂与国际知名度高、产品质量好的美国IDEXX同类试剂对432份临床猪血清进行符合性检测,结果与美国IDEXX试剂的阳性符合率为95.53%,阴性符合率为86.56%,总符合率为91.67%,两种试剂检测结果具有较高的一致性。试验表明,用E2蛋白为包被抗原对猪瘟病毒抗体进行ELISA检测的效果最好,制备的检测试剂可用于临床猪瘟病毒血清抗体的检测,为今后猪瘟病毒抗体ELISA检测试剂的进一步研制奠定了基础。