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抗牛多杀性巴氏杆菌高免卵黄抗体IgY的制备及间接血凝检测方法的建立 总被引：1，自引：0，他引：1
本研究通过自制牛源荚膜血清A型多杀性巴氏杆菌灭活菌苗免疫产蛋鸡,采用卡拉胶结合硫酸铵沉淀法提取卵黄抗体IgY,并采用间接血凝方法检测抗体效价。结果表明,抗原致敏红细胞的最佳浓度是800μg/mL,免疫后第7周抗体效价达到高峰,效价为1：1024,高效价持续5周开始下降,测定提取的IgY浓度为8.258mg/mL,无菌检测及动物安全性实验表明制备的卵黄抗体安全可靠。本研究制备了抗牛多杀性巴氏杆菌卵黄抗体,为防治由荚膜血清A型多杀性巴氏杆菌所致的犊牛肺炎提供了新的手段。 相似文献
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《中国兽医学报》2015,(9):1495-1500
为探讨不同中药多糖对猪蓝耳病(PRRS)卵黄抗体效价的影响,本试验采用水提法制备卵黄水提液,2次盐析法纯化卵黄抗体(IgY),SDS-PAGE法检测精制卵黄抗体的纯度。采用棋盘法确定了抗原最佳包被浓度和酶标二抗最佳工作浓度,建立了PRRS特异性卵黄抗体ELISA检测方法。并用PRRS疫苗4种不同免疫剂量免疫蛋鸡,确定最佳免疫方法。选择桑叶多糖、杜仲多糖、桑叶杜仲多糖、地黄多糖和藿蜂酮注射液饮水给药与PRRS疫苗联合免疫蛋鸡,通过卵黄抗体效价变化考察不同中药多糖的免疫增强作用。结果显示:选择抗原的最佳包被浓度为10-9,酶标二抗的最佳工作浓度为1:5 000,卵黄WFS的最佳稀释度为2-6;鸡肌注0.2mL/羽PRRS疫苗,在免疫后2~6周产生的抗体效价最高;鸡在免疫的同时以4mg/羽的剂量饮水给以桑叶杜仲复方多糖,连续3d,在免疫后的第2~9周卵黄抗体效价均最高,优于其他各组。以上结果表明,桑叶杜仲复方多糖饮水给药配合疫苗一起使用能显著提高PRRS卵黄抗体效价,可作为免疫增强剂使用。 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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张皓淳 《国外畜牧学(猪与禽)》2022,42(1):32-36
为了制备非洲猪瘟病毒(African swine fever virus,ASFV)p30蛋白卵黄抗体,研究卵黄抗体的生物活性。试验构建了p ET-32a-p30原核表达载体,制定了科学的免疫程序和样本采集方法。提取卵黄液,进行二次盐析,得到对应的卵黄抗体。用酶联免疫吸附试验(enzyme-linked immunosorbentassay,ELISA)测定卵黄抗体效价,用免疫印迹（Westernblotting）技术对其生物活性进行鉴定。ELISA试验表明：首次免疫p30蛋白7d后,能够在蛋鸡的血清中检测出抗体（Ig Y）,14d后在卵黄中能够提取到Ig Y,35d后Ig Y效价达到最高值。IgY具备和哺乳动物产生的IgY不同的生物活性,并可具有较高的特异性和稳定性,在免疫学研究中具有重要的价值。利用Ig Y进行实验检测,具有突出优势。通过抗原即ASFV p30蛋白免疫蛋鸡,分离提纯免疫蛋白,检测其生物活性,可用于鉴定非洲猪瘟。 相似文献
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《经济动物学报》2021,25(3):132-139
为建立牛呼吸系统疾病主要病原-牛支原体的快速富集方法,试验采用PCR方法对牛支原体P48基因扩增,构建重组质粒pET32a(+)-P48。将重组质粒进行原核表达并进行纯化。将纯化的牛支原体P48蛋白作为免疫原与弗氏佐剂混合乳化免疫家兔,获得抗牛支原体P48蛋白的高免血清,同时对高免血清进行纯化。将纯化后多克隆抗体与NHS磁性微球进行耦联,制备免疫磁珠。对耦联缓冲液、抗体浓度进行优化。从富集牛支原体的时间、敏感性和特异性三方面对免疫磁珠进行评价。结果表明:成功地扩增出大小约1 407 bp的P48基因并表达出大小约62 ku的可溶性重组蛋白。制备的多克隆抗体的效价高达1∶64 000,且具有很高的特异性;抗体与磁珠耦联的最佳介质为2-吗啉乙磺酸;抗体质量浓度为200μg/mL与磁珠的耦联率最高;免疫磁珠在15 min之内就能够富集到牛支原体,且富集到浓度为1×10-3ccu/mL,具有良好的特异性。说明免疫磁珠技术能够快速富集牛支原体,为分离鉴定牛支原体提供一种新的可行性的方法。 相似文献
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本研究将自制牛支原体和牛源荚膜血清A型多杀性巴氏杆菌灭活菌苗分别免疫产蛋鸡,并检测效价,采用卡拉胶结合硫酸铵沉淀法提取Ig Y,观察抗牛支原体及多杀性巴氏杆菌二联高免卵黄抗体制剂在犊牛中应用的效果,并进行临床应用研究。结果显示,牛支原体在首免后第60d抗体效价达到高峰,经ELISA检测效价为1:128000;牛多杀性巴氏杆菌首免后第50d抗体效价达到高峰,经IHA检测效价为1:1024。临床初步应用表明,二联卵黄抗体对犊牛肺炎的预防和治疗均有一定作用。 相似文献
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制备猪繁殖与呼吸综合征病毒高免卵黄抗体,研究其治疗效果。以猪繁殖与呼吸综合征灭活苗免疫产蛋鸡,采用ELISA方法检测卵黄抗体效价,收集高效价卵黄,采用氯仿抽提和硫酸铵盐析法纯化卵黄抗体。ELISA方法测定收集抗体效价为1∶8 000,测定纯化的抗体浓度为7.85mg/mL,微生物学检验及安全性试验表明制备的卵黄抗体安全可靠,人工感染治愈率和临床应用治愈率分别为100%和89.1%,表明制备的卵黄抗体对猪繁殖与呼吸综合征具有显著的治疗效果。 相似文献
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为分析蛋鸡免疫后卵黄抗体与血清抗体的相关性,试验采集经H5、H9亚型禽流感和新城疫疫苗免疫的黔东南小香鸡、乌蒙乌骨鸡、威宁鸡血清样本和卵黄样本,采用血凝抑制试验进行相关免疫抗体检测。结果:3个地方品种鸡的血清样本和卵黄样本中均可检测出H5、H9亚型禽流感和新城疫免疫抗体,且几何平均效价均大于24以上,达到免疫合格标准,不同地方品种鸡群间差异不显著(P>0.05);免疫后30d,卵黄样本免疫抗体几何平均效价低于血清样本,但差异不显著(P>0.05);免疫后60d,卵黄样本免疫抗体几何平均效价低于血清样本,差异显著(P<0.05)。结果表明:卵黄抗体与血清抗体呈正相关性,卵黄抗体几何平均效价均低于血清抗体。在临床上可用卵黄抗体替代血清抗体进行H5、H9亚型禽流感和新城疫疫苗免疫效果的评价,但应选择在免疫后30d以内。 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。 相似文献
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本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。 相似文献
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以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。 相似文献
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REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air. 相似文献
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo. 相似文献