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从浙江省10多个规模化猪场采集仔猪断奶腹泻病料,分离致病性大肠杆菌。血清学和PCR方法鉴定菌毛型主要为F4和F18。挑取1株致病性和菌毛表达较强的F4阳性菌株,接种改良M inca培养基培养,热抽提法分离菌毛并纯化菌毛蛋白。将50只蛋鸡随机分5组,分别肌注1 mL免疫原/只:A组菌毛蛋白(250μg/只),B、C和D组菌毛蛋白(250,50,10μg/只) 弗氏佐剂,E组为空白对照。21 d加强免疫。定期采血分离血清,水稀释法和饱和硫酸铵盐析法分离卵黄抗体。ELISA法检测血清和卵黄抗体效价。结果表明,F4菌毛蛋白具有良好的免疫原性:在弗氏佐剂的辅助下,二免后21 d抗体效价达最高峰,血清和卵黄抗体效价分别为4.7 lg和4.4 lg,二免后56 d血清和卵黄抗体效价仍维持在4.0 lg;另外二免后21 d,10μg/只免疫剂量与50μg/只及250μg/只相比,诱导产生的卵黄抗体效价之间差异不显著(P>0.05)。 相似文献
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为制备针对一些大型养猪场产肠毒素性大肠杆菌(enterotrxigenic Escherichia coli,ETEC)分离株的特异性卵黄抗体(egg yolk immunoglobulin,IgY),试验对从这些养殖场分离的ETEC分离株菌毛基因类型进行了PCR鉴定,纯化该分离株的菌毛蛋白免疫蛋鸡制备IgY,对该IgY的效价、特异性和体外抑菌效果进行了检测。结果表明,该分离株具有K88和987p 两种菌毛基因,纯化后的分离株菌毛具有较强的免疫原性,经3次免疫后产生的IgY对K88和987p全菌和菌毛的效价可达到1:64 000,分离株菌毛IgY能特异地与K88和987p反应,与K99、F41无交叉反应,5 mg/mL分离株菌毛IgY在体外具有很好的抑菌效果。 相似文献
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细菌CpG—DNA对犬免疫增强效果的研究 总被引：10，自引：1，他引：9
以犬瘟热病毒为模式病毒，将CpG-DNA与铝胶、蜂胶2种对照佐剂分别与犬瘟热病毒灭活疫苗配合，给犬接种疫苗前后用微量中和试验抽查血清中和抗体效价，根据抗体效价高低判断佐剂的免疫增强效果。结果表明，3种佐剂疫苗均能诱导产生特异性抗体，但组间抗体效价有显著差异（P＜0．01），CpG-DNA的免疫增强作用明显高于其它2种佐剂，CpG-DNA所诱导的抗体效价较蜂胶佐剂高5倍，较铝胶佐剂高7倍。 相似文献
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为筛选卵黄抗体的优良佐剂,以3株猪源致病性大肠杆菌(ETEC)菌毛蛋白为模式抗原,分别与6种佐剂制备多价菌毛蛋白疫苗,免疫海兰褐壳蛋鸡。结果显示,3次免疫后,所有试验组蛋鸡产蛋率均有不同程度下降,但SPA、ISCOMs、弗氏佐剂、蜂胶佐剂组的产蛋率下降程度较为轻微;对鸡体血清抗体和制备获得的卵黄抗体的效价检测结果均显示:SPA和ISCOMs组虽较SPB、SPC、弗氏佐剂组抗体水平略低,但持久度较好,且通过对蛋鸡外观形态和注射部位组织变化情况的观察显示,两者不会刺激鸡体热反应或炎症反应,无任何表观副作用。因此,更适合作为蛋鸡制备猪源ETEC卵黄抗体的优良佐剂。 相似文献
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不同佐剂对鸡新城疫疫苗免疫调节作用的影响 总被引：1，自引：0，他引：1
试验以300只AA雏鸡为试验动物,将其随机分为4组,A、B、C组分别接种相同剂量的甲壳素、蜂胶、铝胶佐剂苗,D组为对照组。结果表明,三种佐剂的疫苗在对肉鸡组免疫的第3、4、5、6、7、9、11、13、15、20、25天后,甲壳素疫苗组H I抗体效价显著高于其它组,蜂胶疫苗组H I抗体效价其次,铝胶疫苗组抗体H I效价最低;用三种不同佐剂的疫苗对肉鸡免疫后第26天称其重量,甲壳素佐剂疫苗组免疫后的肉鸡增重效果最好,蜂胶疫苗组居中,铝胶疫苗组最差。 相似文献
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牦牛ETEC灭活疫苗的研制及免疫效力试验 总被引：4，自引：0，他引：4
制备了耗牛ETEC铝胶灭活苗、油佐剂苗和蜂胶灭活苗。分别使用3种佐剂灭活苗进行对比试验。经抗体监测表明在免疫后7天，蜂胶苗产生的抗体效价最高（2^7），随着时间的推移，油苗抗体儿价最高（2^9-10)，于免疫后29日，使用ETEC进行攻毒，免疫兔全部保护，对照兔全部死亡。证明牦牛ETEC3种佐剂灭活疫苗均能有效保护家兔免受ETEC攻击。 相似文献
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本试验将300只AA雏鸡随机分为4组,A、B、C组分别接种相同剂量的甲壳素、蜂胶、铝胶佐剂苗,D组为对照组,从血清免疫学上探讨甲壳素对鸡新城疫疫苗的免疫调节作用,并与蜂胶和氢氧化铝胶佐剂加以比较,以观察不同免疫佐剂对鸡新城疫的免疫调节作用。结果表明,3种佐剂的疫苗在对肉鸡组免疫的第3、4、5、6、7、9、11、13、15、20、25d后,甲壳素疫苗组HI抗体效价显著高于其它组,蜂胶疫苗组HI抗体效价其次,铝胶疫苗组抗体HI效价最低。 相似文献
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IgG antibody response in chickens immunized with F4 fimbriae extracted from local enterotoxigenic Escherichia coli (ETEC) strain was studied during a 98-day immunization period for comparing the efficacy of four adjuvants: Freund' adjuvant, Quil A (QA), propolis and extract from Cochinchina momordica seed (ECMS). For this purpose, chickens were immunized with F4 fimbriae alone or combined with one of the above adjuvants on days 1 and 21. IgG antibody levels in serum and egg yolk (by ELISA) were measured on days 0, 7, 14, 21, 28, 35, 42, 49, 56, 70, 84 and 98. The egg production of each group was also determined during days 1-7 and the following four weeks. The results showed that QA could enhance antibody titre, as good or almost as good as Freund's adjuvant, whereas the titres of ECMS and propolis groups were relatively lower, with the overall order: Freund's adjuvant>QA>ECMS>propolis both in serum and egg yolk. However, the significant decrease of egg production was merely observed in the Freund's adjuvant group. It is concluded that the four adjuvants tested can stimulate immune response to F4 fimbriae in chickens, with Freund's adjuvant giving the best results, followed by QA. 相似文献
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D Ciosek M Truszczy&#x;ski M Jagodzi&#x;ski 《Comparative immunology, microbiology and infectious diseases》1983,6(4):313-319
Different vaccines against Escherichia coli diarrhea of piglets were applied parenterally in pregnant sows at an industrial fattening farm. The following vaccines were used: vaccine No. 1 with non-complete Freund's adjuvant. Tween 80 and Arlacel A, comprising O149:K91,K88; O139:K82; O8:K87,K88; O141:K85,K88; and O64:K? E. coli serotypes; vaccine No. 2 with paraffin oil instead of Freund's adjuvant, comprising the same E. coli serotypes as the vaccine No. 1; stable specific vaccine with 10% aluminium hydroxide, based on E. coli serotypes most frequently isolated from piglets which died at the farm (O149:K91,K88; O8:K87,K88; O20:K17; O64:K?); Gletvax K88 (Wellcome) and NOBI-VAC LT-K88 (Intervet International). The number of piglets which died up to the moment of weaning in comparison to the number of born ones was considered as an indicator of acquired protection. It was found that the most effective in conferring protection against E. coli diarrhea were: vaccine No. 1 and NOBI-VAC. The differences in the mortality rate between piglets originating from sows vaccinated with these vaccines and those from unvaccinated ones were statistically significant (P < 0.05). No significant differences were noted between controls and animals vaccinated with the remaining vaccines. 相似文献
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B Nagy S H?glund B Morein 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1990,37(10):728-738
Iscom (immunostimulating complex) vaccines were prepared to contain K88ab, K88ac, K99 and 987P pili (fimbriae) of enterotoxigenic E. coli bacteria as monovalent or quadrivalent preparations. The iscoms injected into rabbits and into pigs elicited similar or higher immune response in both animal species than the oil adjuvanted vaccine containing about 5 times more of the same pilus protein. It is concluded that inclusion of pili into iscoms results in immunogenic preparations likely worth pursuing for vaccine production against enterotoxic colibacillosis of newborn pigs. The iscoms did not induce local reaction at the injection sites in contrast to the oil adjuvanted vaccines. 相似文献
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Immune responses in chickens against Salmonella typhimurium monitored with egg antibodies 总被引：1，自引：0，他引：1
Three mature hens were immunized with an Aro- mutant of Salmonella typhimurium beginning with a subcutaneous dose in adjuvant followed by two oral boosters. Isotype-specific antibodies were measured in the white and yolk eggs collected weekly over a period of 230 days. Two hens showed a memory response to the first oral booster, with large increases in egg yolk IgG and smaller increases in IgA and IgM antibodies in egg whites. Smaller amounts of IgA and IgM antibodies were found in egg yolks, and a slight increase in IgG occurred in the whites. One hen showed an increase in serum titers of all isotypes against S. typhimurium. The second hen had high serum titers before immunization was started which did not change. The third hen had a high level of IgM in the white of eggs before immunization was started. This hen showed erratic responses in egg white antibodies following immunization, no increase in IgA or IgM in yolks and only a slight increase in IgG, no increase in serum IgG, and was the only hen with a high level of IgM antibody against S. typhimurium in the bile, conditions reflecting a state of oral tolerance. With the exception of this hen, the results showed that IgA and IgM antibodies were aroused in hens by immunization with an avirulent mutant of S. typhimurium, and that these antibodies were present in the white of eggs from immunized hens. 相似文献
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选用4株带有K88、K99、987P、F41粘附素抗原的产肠毒素性大肠杆菌,分别接种于BBL、Minca、Slanetz和Minca培养基进行培养,将培养物加热提取粘附素抗原后,加入油佐剂制成四价亚单位疫苗。经成品检验合格后分别免疫小鼠和怀孕母猪,同时监测怀孕母猪抗体。最后1次免疫后15 d,分别用同源ETEC确定的攻毒剂量攻击。结果显示,经2次免疫后,K88、K99、987P、F41对小鼠的免疫保护率与1次免疫没有显著差异(P>0.05);对仔猪,1次免疫跟2次免疫均可显著地降低腹泻指数(P<0.05),2次免疫与1次免疫没有显著差异(P>0.05)。怀孕母猪免疫1周后(产前23 d)抗体开始上升,第2周(产前16 d)达到高峰。2次免疫后抗体迅速回升,第4周(产前2 d)达到最高峰,产后2 d抗体大幅度下跌,几近免疫前水平。二免母猪所产仔猪发病率明显低于一免母猪(P<0.05)。攻毒保护试验和抗体消长规律的结果表明,制备的仔猪大肠杆菌病K88-K99-987P-F41四价亚单位疫苗具有良好的免疫原性,能有效预防仔猪大肠杆菌病的发生。 相似文献
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本试验旨在研究疫苗、免疫剂量和注射方式对卵黄抗体效价规律的影响,探讨制备抗猪乙型脑炎病毒卵黄抗体蛋鸡的最佳免疫程序。选用无免疫褐羽蛋鸡180只,随机分成18组,每组10只。1、2组均为对照组,注射无菌生理盐水;3~10组采用皮下和肌肉两种注射方式,并依次注射灭活苗0.2、0.5、1.0和1.5 mL;11~18组同样采取两种注射方式,依次免疫相同剂量弱毒苗。各试验组分别于免疫前1 d、免疫后第3、7、10、14、18、21和28天采集当日鸡蛋6枚,提取卵黄抗体,测定效价。试验结果显示,1~6、11、12组卵黄抗体效价均为0,未产生明显免疫应答;7~10组注射灭活苗后7 d,平均卵黄抗体效价达到峰值,抗体持续时间为14 d;13~18组注射弱毒苗后14 d达到最高值,抗体持续时间为21 d;注射剂量相同但注射方式不同的两组之间比较,卵黄抗体效价差异不显著（P>0.05）;相同注射方式,相同疫苗的各试验组间,随着免疫剂量的增加卵黄抗体效价逐渐加强,且差异显著（P<0.05）。以上结果表明,肌肉或皮下两种注射方式,蛋鸡产生明显免疫应答至少需要免疫灭活苗1.0 mL或弱毒苗0.5 mL。比较弱毒苗与灭活苗,灭活苗刺激机体产生的抗体速度较快,但维持时间较短;较少剂量弱毒苗就可刺激机体产生抗体,但速度慢、维持时间长。 相似文献
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The chicken egg as a supply of polyclonal antibodies 总被引：2，自引：0，他引：2
Polyclonal antibodies can be isolated not only from the blood of immunized mammals but also from the egg yolk of immunized chickens. The advantages of this alternative method are: 1) Birds produce antibodies against highly conserved mammalian proteins. 2) The quantity of antigen needed for an efficient immune response is very low (20-30 micrograms). 3) The use of complete Freund's adjuvant leads to long lasting titers of yolk antibodies yielding a total amount of 65 mg specific antibodies per month. 4) The purification of antibodies is simple, inexpensive and quick. Polyethylene glycol precipitation is sufficient to obtain a purity of more than 90%. 5) Chicken antibodies are acid- and heat-resistant and might therefore be orally applied to prevent or to cure infectious intestinal diseases of young animals or humans. 6) Immunization with complete Freund's adjuvant is well tolerated and produces no inflammatory reactions and 7) collecting eggs is, in contrast to bleeding animals, non-invasive. In this review we present both, the method how to produce and to isolate yolk antibodies as well as their possible application in science, diagnosis, prophylaxis and therapy. 相似文献