山羊伪结核棒状杆菌LY20株分离鉴定及病理组织学观察  

Isolation and identification of Corynebacterium pseudotuberculosis from goats and histopathological observation of infected mice

作　　者：王锐鸿 林昶 池雪林[1] 陈仕龙[2] 曾显成[1] WANG Ruihong;LIN Chang;CHI Xuelin;CHEN Shilong;ZENG Xiancheng(College of Bee Science,College of Animal Sciences,Fujian Agricultural and Forestry University,Fuzhou 350002,China;Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agriculture Sciences,Fuzhou 350012,China)

机构地区：[1]福建农林大学,动物科学学院,蜂学学院,福建福州350002 [2]福建省农业科学院,畜牧兽医研究所,福建福州350012

出　　处：《中国兽医学报》2021年第12期2411-2420,共10页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2017YFD0500203);福建农林大学科技创新专项基金资助项目(CXZX2017042,CXZX2017043);福建省自然科学基金资助项目(2018J01595)。

摘　　要：旨在明确福建某羊场引起山羊皮肤脓肿的病原,无菌采集脓液,进行细菌分离培养、培养特性观察、形态学镜检、生化试验及16S rRNA进化树分析,结果分离到1株细菌,经鉴定确定为伪结核棒状杆菌,命名LY20株。毒力因子检测显示,分离菌携带PLD、NanH、FagA、FagB、FagC、FagD、OppA、OppB、OppC、OppD、OppF共11个毒力因子。PLD全基因进化树分析,结果伪结核棒状杆菌分成羊型和马型2个独立分支,分离菌与羊型聚在同一分支,亲缘性最近,与另一个分支马型亲缘性较远。核苷酸和氨基酸同源性比较显示,分离菌与羊型伪结核棒状杆菌同源性分别为99.9%和99.7%,与马型伪结核棒状杆菌的同源性分别为98.1%~98.2%和97.4%。用4.4×10^(8)CFU/mL菌液,0.2 mL/只,腹腔接种小鼠,48 h内死亡率达100.0%,表现出较强的致病性;皮下接种小鼠,接种部位出现脓肿、破溃后流出淡黄色脓液。病理切片观察,可见肝脏淤血,脾淋巴细胞坏死,肺脏出血及中性粒细胞浸润,肾小管上皮细胞坏死,十二指肠和直肠上皮细胞脱落、间质出血、淋巴细胞浸润;皮肤化脓坏死性肌炎,肉芽组织增生。药敏结果显示,分离菌对环丙沙星、恩诺沙星等28种药物敏感,对阿米卡星、卡那霉素等7种药物耐药。研究结果表明,本次山羊皮肤脓肿为伪结核棒状杆菌感染所致,且小鼠病理组织学变化主要表现为出血、坏死及化脓性炎症。In order to identify the pathogeny of subcutaneous abscess of goats in a goat farm in Fujian Province,the pus was collected aseptically for bacterial isolation,morphological microscopic examination,culture characteristics observation,biochemical test and 16 S rRNA sequencing analysis.A strain of bacteria was isolated and identified as Corynebacterium pseudotuberculosis,it was named LY20 strain.The virulence gene test showed that the isolate carries at least 11 virulence genes,such as PLD,NanH,FagA,FagB,FagC,FagD,OppA,OppB,OppC,OppD and OppF.Phylogenetic tree analysis of PLD gene showed that Corynebacterium pseudotuberculosis was divided into two independent clades:Ovis and Equine,and the isolate was clustered in the same branch of Ovis with the closest genetic relationship,but far from Equine.The nucleotide and amino acid homology of the isolate was 99.9%and 99.7%with Corynebacterium pseudotuberculosis of Ovis,98.1%-98.2%and 97.4%with Corynebacterium pseudotuberculosis of Equine.The mice were intraperitoneal inoculated with 0.2 mL of 4.4×10^(8)CFU/mL bacterial solution,and the inoculated mice had a mortality rate of 100.0%within 48 h,showing that the isolate had strong pathogenicity.Pathological sections showed liver congestion,splenic lymphocyte necrosis,pulmonary hemorrhage and neutrophil infiltration,renal tubular epithelial cell necrosis,duodenal and rectal epithelial cell abscission,interstitial hemorrhage and lymphocyte infiltration,skin septic necrotizing myositis and granulation tissue hyperplasia.Drug sensitivity test showed that the isolate was sensitive to 28 kinds of drugs such as ciprofloxacin and enrofloxacin,and resistant to 7 kinds of drugs such as amikacin and kanamycin.The results showed that the goat skin abscess was caused by Corynebacterium pseudotuberculosis infection,and the main histopathological changes of mice were hemorrhage,necrosis and suppurative inflammation.

关 键 词：山羊 伪结核棒状杆菌 分离鉴定 药敏特性 毒力因子 病理学观察 

分 类 号：S855.1[农业科学—临床兽医学] R535[农业科学—兽医学]