陕南白山羊伪结核棒状杆菌分离鉴定及灭活铝胶疫苗研制

发布时间：2018-12-07 10:06

【摘要】：羊伪结核病(ovine pseudotuberculosis)又称羊干酪样淋巴结炎(caseous lymphadenitis),是由伪结核棒状杆菌(Corynebacterium pseudotuberculosis)所引发的人畜共患慢性病,绵羊和山羊多发,可造成患病羊消瘦、生产性能低下和产死胎,对养羊业的危害较大。石泉县位于陕西省安康市以西,北依秦岭、南枕巴山,非常适合陕南白山羊养殖,一直以来是安康市陕南白山山羊养殖大县之一,仅2014年出栏陕南白山羊64119只。近年来石泉县养羊户反映所养的陕南白山羊群中有部分羊只在前肢上部出现一种“脓包”,多种方法治疗效果不佳,在羊群中持续存在,造成了较大的经济损失。本文对该病进行了发病情况调查、诊断和防治试验,获得了以下结果。(1)临床发病调查结果表明,随机抽取的饲养量在50只以上的圈养和散养大户各10个,圈养公羊、母羊和羔羊“脓包”平均发病率分别为46.4%、38.1%和1.5%,散养公羊、母羊和羔羊“脓包”平均发病率分别为69.6%、59.1%和2.9%。(2)从随机采取的患病羊只病料30份中分离培养到1株疑似伪结核棒状杆菌。应用细菌16S rRNA通用引物进行PCR扩增和序列分析表明,该分离菌与羊伪结核棒状杆菌16S rRNA序列的同源性为99.5%,确定分离菌株为羊伪结核棒状杆菌,确诊该病为羊伪结核病。(3)药敏试验结果表明,该菌对环丙沙星、头孢曲松、红霉素、卡那霉素、头孢噻肟、妥布霉素、强力霉素、庆大霉素、氯霉素等抗菌药物高度敏感;对四环素、利福平、链霉素等中度敏感。临床治疗试验结果表明,利福平和头孢噻肟钠经过皮下注射给药方式,对羊伪结核病治疗效果较好。(4)应用分离到的羊伪结核棒状杆菌研制了灭活铝胶疫苗,在安全性试验和无菌试验证明疫苗正常的情况下,进行小规模田间试验的结果表明,在一年的时间里,接种疫苗的30只羊中只有1只羊患上了羊伪结核病,发病率在3.3%,保护率为96.7%,而注射无菌生理盐水的对照组30只羊中有23只羊患上了羊伪结核病,发病率为76.7%。说明研制的疫苗对山羊伪结核病有很好的预防效果。
[Abstract]:Sheep pseudotuberculosis (ovine pseudotuberculosis), also called goat caseous lymphadenitis (caseous lymphadenitis), is a chronic zoonotic disease caused by Corynebacterium pseudotuberculosis (Corynebacterium pseudotuberculosis). Sheep and goats have multiple diseases, which can lead to emaciation, low productivity and stillbirth in sheep. The harm to sheep industry is great. Shiquan County is located in the west of Ankang City, Shaanxi Province, north according to the Qinling Mountains, Nanxingba Mountain, very suitable for the breeding of white goats in southern Shaanxi, has been one of the largest counties of Ankang City, Shannan White Mountain goat breeding, only in 2014, 64119 white goats in southern Shaanxi. In recent years, Sheep farmers in Shiquan County have reported that some of the sheep in the Southern Shaanxi White Goat herd have a kind of "abscess" in the upper part of the forelimb. The following results were obtained from the investigation, diagnosis and prevention and treatment of the disease. (1) the results of the clinical investigation showed that 10 large captive sheep and 10 free-range large households were randomly selected, and the male sheep were kept in captivity. The average incidence of abscess in ewe and lamb was 38.1% and 1.5%, respectively. The average incidence of sheep, ewe and lamb was 69.6%, respectively. 59.1% and 2.9% respectively. (2) A suspected pseudotuberculosis corynebacterium was isolated from 30 samples of diseased sheep. The results of PCR amplification and sequence analysis using 16s rRNA universal primers showed that the homology of 16s rRNA sequence between this isolate and Corynebacterium tuberculosis was 99.5, and the isolated strain was identified as Corynebacterium pseudotuberculosis. (3) the drug sensitivity test showed that the strain was resistant to ciprofloxacin, ceftriaxone, erythromycin, kanamycin, cefotaxime, tobramycin, doxycycline, gentamicin, Chloramphenicol and other antimicrobial agents are highly sensitive; Moderately sensitive to tetracycline, rifampicin, streptomycin, etc. The results of clinical treatment showed that rifampicin and cefotaxime sodium were injected subcutaneously to treat pseudo-tuberculosis in sheep. (4) inactivated aluminum gel vaccine was developed by using isolated Corynebacterium pseudotuberculosis in sheep. When the safety test and aseptic test proved that the vaccine was normal, the results of small-scale field trials showed that only one sheep out of 30 vaccinated sheep developed pseudo-tuberculosis in one year, and the incidence rate was 3.3%. The protective rate was 96. 7%, while 23 of the 30 sheep in the control group were infected with pseudotuberculosis, the incidence rate was 76. 7%. The results showed that the vaccine had a good preventive effect on goat pseudopulmonary tuberculosis.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S858.27

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