1株猪源芳香黄杆菌的分离鉴定及生物学特性研究  

Isolation,identification and biological characteristics of a Myroides sp.strain

作　　者：徐松平 梁海英[1] 吴学祥[1] 曾智勇[1] 汤德元[1] 王彬[1] 徐玉 祝羊 黄二素 张婧旭 万娟 XU Song-ping;LIANG Hai-ying;WU Xue-xiang;ZENG Zhi-yong;TANG De-yuan;WANG Bin;XU Yu;ZHU Yang;HUANG Er-su;ZHANG Jing-xu;WAN Juan(College of Animal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025

出　　处：《中国预防兽医学报》2021年第6期597-601,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：“水芋香猪绿色养殖关键技术研究及示范”黔科合支撑[2020]1Y033号;“猪呼吸道疾病综合征多重PCR的建立及防治研究”黔科合支撑[2019]2276号项目。

摘　　要：为确定贵州省仁怀某养殖场出现咳嗽、发烧、间歇性腹泻猪的发病原因,本研究从无菌采集的患病猪心脏、肺脏等组织样品中分离到1株病原菌,对其纯化培养后进行形态学检测,发现该菌落在TSA培养基、LB培养基均长出肉眼可见的淡黄色、表面光滑、有芳香味的圆形菌落;在血平板上长出无溶血现象的乳白色、表面光滑的圆形菌落;继续培养12 h后发现,LB培养基上菌落呈棕色,TSA培养基和血平板上的菌落则无变化。革兰氏染色镜检可见红色的长杆菌。进一步生化鉴定结果显示该分离菌的生化特性与芳香黄杆菌符合。对细菌16S rRNA基因PCR扩增分析并构建进化树,结果显示,分离菌株与芳香黄杆菌OM424株(MK034244)基因序列同源性为99%以上,且与芳香黄杆菌OM424株(MK034244)和芳香黄杆菌TH-19株(NR133793)属于同一个遗传进化分支,遗传关系较近。以上结果可确定该分离菌为芳香黄杆菌,将其命名为RH564。药敏试验结果显示,RH564株仅对红霉素、麦迪霉素、万古霉素敏感,表明芳香黄杆菌对多种抗生素耐药性较强。以4×10^(8)cfu/只剂量反复涂抹实验鼠伤口,进行小鼠致病性试验,结果显示,感染组小鼠均出现临床症状,甚至死亡,并且可从死亡小鼠的肺脏、肝脏中检测到该分离菌株,表明芳香黄杆菌RH564株具有一定的致病性。本研究首次在中国分离出猪源芳香黄杆菌,为临床上芳香黄杆菌病的预防和治疗奠定了基础。To determine the causes of cough,fever,and intermittent diarrhea in a pig herd in Renhuai,Guizhou province,this study isolated a strain of pathogenic bacteria from the aseptically collected heart and lung tissues of diseased pigs.After purification,morphological examination showed that the colony on TSA medium and LB medium both grew pale yellow,smooth and aromatic round colonies visible to the naked eye,and a milky white,smooth surface without hemolysis on the blood plate.After 12 hours of continuous culture,it was found that the colonies on the LB medium were brown,while there was no change on the TSA medium and blood plate.Gram staining and microscopic examination showed red long bacilli.Further biochemical identification results showed that the biochemical characteristics of the isolates were consistent with Myroides sp..The results of bacterial 16 S r RNA gene PCR amplification and homology analysis and construction of a genetic tree showed that the gene sequence homology between the isolate and Myroides sp.OM424(MK034244)was more than 99%,and the isolate had a close genetic relationship with Myroides sp.OM424(MK034244)and Myroides sp.TH-19(NR133793),belonging to the same genetic evolutionary branch.From the above results,it can be determined that the isolate is Myroides sp.,and named as RH564.The results of the drug susceptibility test showed that RH564 was only sensitive to erythromycin,midenomycin,and vancomycin,indicating that it had strong resistance to multiple antibiotics.The results of pathogenicity test in mice showed that the infected mice all showed clinical symptoms and even died;the isolate could be detected by PCR in the lungs and livers of dead mice.It shows that the RH564 has certain pathogenicity.This study has isolated swine origin Myroides sp.for the first time in China,which lays a foundation for the prevention and treatment of Myroides sp.

关 键 词：芳香黄杆菌 分离鉴定 药敏试验 人畜共患 

分 类 号：S852.61[农业科学—基础兽医学]