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

羊干酪性淋巴结炎又称羊伪结核病,是由伪结核棒状杆菌感染引起的一种人畜共患慢性接触性传染病,对山羊危害最为严重,目前尚无有效治疗措施。陕西省石泉县陕南白山羊饲养量较大,羊群也存在伪结核棒状杆菌的感染,造成较大的经济损失。为筛选对当地伪结核棒状杆菌敏感的药物和研制灭活疫苗,本研究从疑似伪结核病患羊采取多份新鲜的脓液,进行细菌分离鉴定;对获得的1株分离菌进行药物敏感性试验并用其研制灭活疫苗。结果显示,分离菌在固体培养基上形成白色、干燥、扁平、不透明、边缘整齐的中等大小菌落,革兰染色阳性,经16SrRNA检测和序列分析,证实分离到的菌株为羊伪结核棒状杆菌。药物敏感性试验结果显示,该分离菌对环丙沙星、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、氯霉素高度敏感;对四环素、利福平、链霉素中度敏感。以该分离菌为种子菌制备的灭活铝胶疫苗无菌检验合格,试验接种山羊无不良反应,目前已经在采样羊场进行临床应用,进一步评价免疫效果。

Isolation and Identification of Corynebacterium pseudotuberculosis from White Goat in South Shaanxi and Development of an Inactivated Vaccine with Aluminum Hydroxide Adjuvant

Caseous lymphadenitis also known as ovine pseudotuberculosis,is a chronic zoonotic infectious disease caused by Corynebacterium pseudotuberculosis .This disease is harmful to goat especially,and there is not yet an effective treatment to prevent this disease.White goat is a main breed in Shiquan county of south Shaanxi,Corynebacterium pseudotuberculosis is an important pathogen in goat population and causes large economic losses in recent years.In order to screen drugs that are sensitive to local Corynebacterium pseudotuberculosis and develop an inactivated vaccine,the fresh pus were collected from the suspected pseudotuberculosis goats,the bacteria were isolated and identified from those samples.And then,drug sen-sitivity test was carried out with 1 isolate,and then an inactivated vaccine with aluminum hydroxide adju-vant was developed based this isolate.The results showed that the isolated bacteria formed a medium sized white,dry,flat and opaque,neat edge colony at solid agar,showing Gram stain positive.The isolate was confirmed as Corynebacterium pseudotuberculosis by analysis of 1 6 S rRNA sequences.Susceptibility test results showed this isolate was highly sensitive to ciprofloxacin,ceftriaxone,erythromycin,kanamycin,ce-fotaxime,moderately sensitive to tobramycin,doxycycline,gentamicin,chloramphenicol;tetracycline,rif-ampicin,streptomycin.Subsequently,an activated vaccine was developed with aluminum hydroxide adjuvant based on this isolate,no live bacterium was detected in this vaccine.The inactivated vaccine was injected to experimental goats in those herds that the samples were collected,and no abnormal reactions were ob-served in the goats.Clinical application also was carried out in goat herd and the immune effects will be e-valuated later.This study provided an experimental basis for effective drugs and a choice of inactivated vac-cine for preventing and controlling caseous lymphadenitis.