山羊副结核病的诊断和分型鉴定  

Diagnosis and Genotyping Identification of Paratuberculosis in Goats

作　　者：孟维康 赵维宏 李梦园 马薇 丁玉林[1,2,3] 吉林台 赵丽 刘永宏 MENG Weikang;ZHAO Weihong;LI Mengyuan;MA Wei;DING Yulin;JIRINTAI S;ZHAO Li;LIU Yonghong(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010000,China;Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease,Ministry of Agriculture and Rural Affairs,Hohhot 010000,China;National Center of Technology Innovation for Dairy,Hohhot 010000,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010000 [2]农业农村部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010000 [3]国家乳业技术创新中心,内蒙古呼和浩特010000

出　　处：《中国兽医杂志》2024年第5期56-62,共7页Chinese Journal of Veterinary Medicine

基　　金：国家乳业技术创新中心项目(2023-JSGG-5);内蒙古自治区自然科学基金项目(2022MS03023);国家自然科学基金项目(32260887)。

摘　　要：为了对1起山羊感染副结核分枝杆菌(MAP)的病例进行诊断,并对其感染的MAP菌株进行分型鉴定,本试验通过病史调查、尸体剖检观察和组织病理学检查对其进行初步诊断,通过聚合酶链式反应(PCR)扩增MAP特异性IS900和IS1311基因,并在测序后进行菌株鉴定、序列分析和IS900基因系统进化树构建。结果显示,病羊极度消瘦,血液稀薄;空肠和回肠肠壁增厚、黏膜面形成皱襞呈脑回样;肠系膜淋巴结肿大,呈灰黄色,切面可见灰白色病灶。肠道组织苏木素-伊红染色可见空肠、回肠黏膜固有层和黏膜下层有大量的上皮样细胞和淋巴细胞增生;肠系膜淋巴结皮质区可见大量上皮样细胞增生,副皮质区有淋巴细胞增生;抗酸染色可见聚集在上皮样细胞内红染的小杆菌。PCR扩增IS900和IS1311基因产物均与目的基因片段预期大小一致;基因序列分析和IS900基因系统进化树显示,与本试验所得分离株同源性最高的参考序列均为Ⅱ型MAP,且同源性均高于98%。综上所述,该羊场发病羊确诊患副结核病,感染菌株为Ⅱ型MAP。本试验为副结核病的诊断和防治提供了参考。In order to diagnose a case of goats infected with Mycobacterium avium subsp.paratuberculosis(MAP)and identify the genotype of the MAP strain causing infection,so as to provide scientific basis for the diagnosis and control of sheep paratuberculosis,this study conducted a preliminary diagnosis through medical history investigation,post-mortem examination,and histopathological examination.MAP-specific IS 900 and IS 1311 genes were amplified by polymerase chain reaction(PCR),and strain identification,sequence analysis,and construction of IS 900 gene phylogenetic tree were performed after sequencing.The results showed that the diseased goats were extremely emaciated with thin blood.The walls of the jejunum and ileum were thickened,and the mucosal surface formed folds resembling cerebral convolutions.The mesenteric lymph nodes were enlarged and gray-yellow,with gray-white lesions visible on the cut surface.Histopathological examination of intestinal tissues stained with hematoxylin-eosin revealed proliferation of epithelioid cells and lymphocytes in the lamina propria and submucosa of the jejunum and ileum.The cortical area of the mesenteric lymph nodes showed proliferation of epithelioid cells,while the paracortical area showed lymphocyte proliferation.Acid-fast staining revealed acid-fast bacilli clustered within epithelioid cells.PCR amplification of IS 900 and IS 1311 genes yielded products consistent in size with the target gene fragments.Sequence analysis and construction of IS 900 gene phylogenetic tree revealed that the highest homology reference sequences obtained in this study were of type II MAP,with homology exceeding 98%.In conclusion,the diseased goats in the goat farm were diagnosed with paratuberculosis,and the infecting strain was identified as type II MAP.This study provides reference for the diagnosis and control of paratuberculosis.

关 键 词：副结核病 山羊 诊断 基因分型 

分 类 号：S858.27[农业科学—临床兽医学]