【作者】 邢福珊；

【导师】 张彦明；

【作者基本信息】 西北农林科技大学 ， 预防兽医学， 2003， 硕士

【摘要】 绵羊和山羊干酪性淋巴结炎（caseous lymphadenitis in sheep and goat ，简称 CLA）也称绵羊和山羊假（伪）结核病（pseudotuberculosis in sheep and goat），是由假（伪）结核棒状杆菌（Corynebacterium pseudotuberculosis）引起的一种人畜共患的慢性传染病。伪结核棒状杆菌能使绵羊、山羊、骆驼、马、牛等多种动物致病，发病特征是受害动物的淋巴结肿大，呈脓性干酪性坏死；有的在肺、肝、脾和子宫角发生大小不等的结节，内含淡黄色干酪样物质。病羊消瘦，生产性能下降，患病孕羊产出死胎，严重者死亡。由于该病发病缓慢，致死率低，所以常被人们忽视，发病后用抗生素治疗效果也不佳，所以现在各个国家都对该病的预防、治疗、检测都重视起来。 分别从杨凌 2 个羊场的 2 只羊的肩前淋巴结脓肿内分离了 2 株菌，经分离鉴定为假（伪）结核棒状杆菌，记为菌株 A、菌株 B。购买 16 只健康山羊，随机分为 4 组，菌株 A 复制病例 1 组，菌株 B 复制病例 1 组，标准菌株接种 1 组，空白对照 1 组。从杨凌养羊场和养羊农户采集了 100 份羊的血清，作为待检血清。用标准菌株制成灭活苗，免疫 1 只山羊，采集其血液，分离血清，即为阳性血清。将临床健康和菌体凝集抑制试验阴性羊的剖腹产胎儿的血清作为阴性血清。 将标准菌株（ATCC19410 株）接种灭菌好的含 0.2%吐温-80 的马丁肉汤中，37℃培养 7d 后制成凝集抑制试验抗原，采集 4 组试验组羊的血清，做菌体凝集抑制试验，只有空白对照组对该菌原有的凝集不抑制，其他 3 组均能抑制凝集。用待检血清做菌体凝集抑制试验，同时设置阴性对照和阳性对照，若假（伪）结核棒状杆菌原有的凝集被抑制，则试验阳性，否则为阴性。 将标准菌株（ATCC19410 株）接种于灭菌好的含 0.2%吐温-80 的马丁肉汤中，振荡培养 3d，离心，上清液用滤器滤过，制成外毒素抗原，通过 ELISA 酶标抗体和阳性血清最适浓度的选择、外毒素抗原和阳性血清最适浓度的选择试验，选用适当的抗原、抗体、酶标抗体浓度，设立空白孔、阳性孔、阴性孔做待检血清的 ELISA 试验，得出阳性判定值，若待检血清 OD 值大于阴性血清的临界值，则判为阳性，否则判为阴性。 用建立的菌体凝集抑制试验和 ELISA 方法检测 100 份待检羊血清，得到了相同的结果,检测到阳性羊 4 只，阳性检出率为 4％。更多还原

【Abstract】 Caseous lymphadenitis in sheep and goat (CLA) caused by CorynebateriumPseudotuberlosis which could infect sheep, goat, camelpa, horse, bovine and so on, is akind of zoonotic and chronic infectious. Infected sheep appeared lymphondusswell,suppurative and caseous necrosis,emerging tubercles in lungs,spleen,uterus horn.Reduce of production and occur of dead fetus in infecred pregnant sheep.The occur of thedisease was slow and lethal rate low.After infection,effect of antibictic treatment to it wasnot good.So every country was increasing attaching importance to prevention theraphyexamina- tion of the disease. 2 bacteria were respectively separated from 2 sheep of Yangling’s 2 sheep-farms inpreshoulder lymphonolus abscess,and identified as Corynebate- riumPseudotuberlosis,marked as A and B.16 healthy goats were respectively divided into 4groups.3 groups were respectively infected bacA bacBand standard CorynebateriumPseudotuberlosis,and 4th group as control group.100 sheep sera were collected fromYangling as samples.Before one month vaccining one goat by sterilized vaccine made bystandard Corynebaterium Pseudotuberlo- s is,we collected serum as negative serum. Weuse clinical health goat seras or somatic inhibited agglutination test negative goat cesareansection fetus seras as negative seras. Corynebaterium Pseudotuberlosis ATCC19410 was seeded into sterilized Martin-meatextract including 0.2% tween-80,cultured in 37℃.7 days later, harvested and made intoantigen of agglutination-inhibition test. Sera were collected from 4 groups,except controlgroup sera,other 3 groups sera all inhibited agglutination of the bacterium.Collectingexaminee’s sera ,we did agglutination-inhibition test meanwhile setting negative andpositive contrast.If Corynebaterium Pseudotuberlosis’agg; utination were inhibited, itseemed as positive,other than as negative. Corynebaterium Pseudotuberlosis ATCC19410 was seeded into sterilized Martin-meatextract including 0.2% tween-80,cultured vibrately for 3 days, centifugaled andfiltered,made into exotoxin antigen.Selecting appropriate consistency of antigen,antibodyand enzyme-labelled,we did ELISA for examine sera and obtained positive determiningvalure,setting blank, positive and negative well.If OD valure of samples were more than<WP=6>positive determining valure,it was positive,other than it was negative. We use inhibited agglutination and ELISA examine 100 seras , results in 4 positiveseras , positive percent is 4％.更多还原