摘要
对安徽省凤阳县某生猪养殖场送检的病死猪肝脏样品进行病原的分离与鉴定。将病死猪肝脏组织接种至血琼脂培养基上进行病原菌的分离纯化、染色镜检、生化试验,以及16S rRNA基因的PCR扩增和测序分析。结果显示,分离菌在血琼脂培养基上呈灰白色、圆形小菌落,菌落周围有草绿色的不完全溶血现象;染色镜检发现分离菌为革兰氏阳性的二联或四联球菌;生化试验表明分离菌对葡萄糖、乳糖和蔗糖均能分解产酸不产气,甲基红试验为阳性,二乙酰和吲哚试验为阴性;PCR测序分析表明,分离菌与猪源绿色气球菌719菌株的16S rRNA基因序列(GenBank登录号为:MN513224)的相似性高达99.86%,与GenBank中的10株绿色气球菌参考菌株的同源性为99.2%~99.3%,与绿色气球菌的多个菌株位于同一大分支上,遗传亲缘关系较近。结果表明,从养殖场送检的病死猪肝脏组织中分离鉴定出1株绿色气球菌AHFY菌株,为该病的临床诊断及防控提供依据。
The study aimed to isolate and identify a strain of Aerococcus viridans from the liver tissues of dead pigs in one farm of Fengyang country in Anhui province by using bacterial isolation and culture,Gram staining,bacterial morphology observation,biochemical assay,PCR and sequencing based on 16S rRNA gene.The morphology observation showed that a Gram-positive diplococcus or detragenococcus strain was isolated,and the isolated strain was grayish-white,round and small colonies with a grass-green incomplete hemolysis around on the blood agar plates.Biochemical assay showed that the reactions of glucose,lactose,sucrose and methyl red were positive,and the reactions of diacetyl and indole were negative.16S rRNA gene sequencing analysis showed that,the similarity of the isolated strain and strain derived Aerococcus viridans published in NCBI(GenBank ID:MN513224)is as high as 99.86%.In addition,the homology of 16S rRNA gene sequence of the isolated strain is ranged from 99.2%to 99.3%compared to that of ten reference strains of Aerococus viridans deposited in NCBI.Phylogenetic analysis showed that the isolated strain belongs to the same genus and grouped in the same branch as some reference strains of Aerococus viridans.In conclusion,an Aerococcus viridans strain(AHFY)was isolated and identified from the liver tissue of dead pigs.
作者
郭伟娜
路振香
陈雪龙
刘畅
张留君
赵磊
GUO Wei-na;LU Zhen-xiang;CHEN Xue-long;LIU Chang;ZHANG Liu-jun;ZHAO Lei(College of Animal Science,Anhui Science and Technology University,Fengyang,Anhui,233100,China)
出处
《动物医学进展》
北大核心
2023年第2期141-144,共4页
Progress In Veterinary Medicine
基金
安徽省教育厅自然科学研究重点项目(KJ2020A0087)
安徽科技学院稳定人才项目(DKWD201801)
安徽省教育厅自然科学研究重点项目(KJ2016A822)
安徽省兽医学重点学科建设项目(AKZDXK2015A04)。
关键词
猪
绿色气球菌
分离
鉴定
16S
rRNA
swine
Aerococcus viridans
isolation
identification
16S rRNA
