山羊源伪结核棒状杆菌的分离鉴定及其毒力评价  

Isolation,identification and virulence evaluation of goat-derived Corynebacterium pseudotuberculosis

作　　者：崔宁 党光辉 鹿萍 郭若男 姜艳艳 陈凡若 刘思国[2] CUI Ning;DANG Guanghui;LU Ping;GUO Ruonan;JIANG Yanyan;CHEN Fanruo;LIU Siguo(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China;State Key Laboratory of Animal Disease Control and Prevention,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052 [2]中国农业科学院哈尔滨兽医研究所动物疫病防控全国重点实验室,哈尔滨150069

出　　处：《黑龙江畜牧兽医》2024年第10期65-70,114,115,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(32002256,32273005)。

摘　　要：为了确定齐齐哈尔市某山羊养殖场疑似患有伪结核病病羊的病原及其毒力,试验从不同病羊的皮下脓包中采集脓汁病料3份,通过细菌的分离与纯化、生化鉴定、16S rDNA基因的PCR扩增与序列分析确定病原,并通过致病性试验对分离株的毒力进行评价。结果表明:共得到3株分离株,其菌落形态和生化特性菌与伪结核棒状杆菌相符,分别命名为CP045、CP073、CP969;分离株CP045、CP073、CP969的16S rDNA基因序列与伪结核棒状杆菌参考株的相似性分别高达98.8%、100%、99.8%,进一步确定3株分离株均为伪结核棒状杆菌。注射分离株CP073的3只小鼠中有2只于注射后36~48 h陆续死亡,1只精神极度沉郁、食欲不振;注射分离株CP045、CP969的3只小鼠于注射后5 d内均未见死亡,但状态不佳。注射分离株CP045、CP073、CP969的小鼠肝脏均不同程度出血、肿大,脾脏轻微出血;注射分离株CP073的小鼠皮肤表面有黄色脓汁渗出,皮下有黄白色玉米粒大小的结核样脓肿;注射分离株CP045和CP969的小鼠皮下未见明显脓肿。说明该山羊养殖场疑似患有伪结核病的病羊感染了伪结核棒状杆菌,分离到的3株伪结核棒状杆菌毒力不同,其中分离株CP073可引起小鼠死亡,毒力较分离株CP045、CP969强。In order to determine the pathogen and its virulence in goats suspected of having pseudotuberculosis in a goat farm in Qiqihar City,in the experiment,three pus samples were collected from subcutaneous pustules of different diseased sheep;the pathogens were determined by isolation and purification of bacteria,biochemical identification,PCR amplification of 16S rDNA gene sequences and sequence analysis;and the virulence of the isolates was evaluated by pathogenicity test.The results showed that a total of three isolates were obtained,and their colony morphology and biochemical characteristics were consistent with those of Corynebacterium pseudotuberculosis,which were named CP045,CP073 and CP969,respectively;the 16S rDNA gene sequences of the isolates CP045,CP073 and CP969 showed a high degree of similarity with the reference strain of Corynebacterium pseudotuberculosis as high as 98.8%,100%,and 99.8%,respectively,which further determined that all three isolates were Corynebacterium pseudotuberculosis.Two of the three mice injected with isolate CP073 died within 36-48 h,and one was extremely depressed and had a poor appetite,but did not die.All three mice injected with isolates CP045 and CP969 did not die within 5 d,but were in poor condition.The liver of mice injected with isolates CP045,CP073 and CP969 was hemorrhagic and enlarged to various degrees,and the spleen was slightly hemorrhagic.The mice injected with isolate CP073 had yellow pus oozing from the skin surface and yellowish-white corn grain-size tuberculosis-like abscesses under the skin;the mice injected with isolates CP045 and CP969 did not have obvious abscesses under the skin.The results suggested that sick goats suspected of having pseudotuberculosis in this goat farm were infected with Corynebacterium pseudotuberculosis;the three isolated strains of Corynebacterium pseudotuberculosis differed in virulence,among which the isolate CP073 could cause the death of mice,and its virulence was stronger than those of the isolates CP045 and CP969.

关 键 词：伪结核棒状杆菌 伪结核病 分离 鉴定 16S rDNA 致病性试验 生化鉴定 

分 类 号：S855.1[农业科学—临床兽医学]