中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (9): 3014-3021.doi: 10.16431/j.cnki.1671-7236.2020.09.037

• 基础兽医 • 上一篇    下一篇

藏猪产色葡萄球菌分离鉴定及生物学特性研究

赵霞玲1,2, 肖琦2, 钱雯娴1,2, 温东旭1, 严明帅1, 吕立新2, 牛家强1, 何孔旺2   

  1. 1. 西藏农牧学院动物科学学院, 西藏高原动物疫病研究自治区高校重点实验室, 林芝 860000;
    2. 江苏省农业科学院兽医研究所, 农业农村部兽用生物制品工程技术重点实验室, 南京 210014
  • 收稿日期:2020-03-06 出版日期:2020-09-20 发布日期:2020-09-16
  • 通讯作者: 牛家强, 何孔旺 E-mail:463036297@qq.com;kwh2003@263.net
  • 作者简介:赵霞玲(1995-),女,四川广元人,硕士生,研究方向:高原动物传染病学,E-mail:1490241436@qq.com
  • 基金资助:
    国家公益性行业(农业)科研专项(201303041);西藏农牧学院研究生创新计划资助项目(YJS2019-21)

Isolation,Identification and Biological Characteristics of Staphylococcus chromogenes from Tibetan Pigs

ZHAO Xialing1,2, XIAO Qi2, QIAN Wenxian1,2, WEN Dongxu1, YAN Mingshuai1, LYU Lixin2, NIU Jiaqiang1, HE Kongwang2   

  1. 1. Provincial Key Laboratory of Tibet Plateau Animal Epidemic Disease Research, College of Animal Science, Tibet Agricultural and Animal Husbandry University, Linzhi 860000, China;
    2. Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
  • Received:2020-03-06 Online:2020-09-20 Published:2020-09-16

摘要: 为了解西藏林芝市健康藏猪的葡萄球菌感染情况,对从西藏林芝市工布江达县巴河镇屠宰场、林芝市巴宜区西藏农牧学院教学实习牧场、林芝市巴宜区屠宰场采集的232份样品进行了细菌分离培养与鉴定,并对1株分离菌株进行了较为系统的生物学特性研究。流行病学调查结果显示,在232份样品中,共检出73份葡萄球菌阳性,阳性率为31.47%,其中在西藏林芝工布江达县巴河镇屠宰场检出16份阳性,检出率为23.89%(16/67);林芝市巴宜区西藏农牧学院牧场检出43份阳性,检出率为34.40%(43/125);林芝市巴宜区屠宰场检出14份阳性,检出率为35.00%(14/40);分离菌经PCR鉴定、基因测序、生化鉴定,证明分离到5株产色葡萄球菌,对其中一株(Sa LZ1)进行了系统的研究。Sa LZ1菌株在琼脂培养基上形成圆形凸起、边缘整齐、表面光滑、不透明的瓷白色菌落,革兰氏染色为阳性球菌;PCR测序结果显示,分离株与产色葡萄球菌(GenBank登录号:MG576253.1)同源性最高;生化鉴定结果显示,其对血清菊糖、甘露醇、葡萄糖、蔗糖、乳糖等表现为阳性,对阿拉伯糖、马尿酸钠、棉子糖、木糖、尿素等表现为阴性,符合产色葡萄球菌的生化特性;Sa LZ1在接种后5~10 h处于对数生长期;小鼠致病性试验显示,当Sa LZ1菌株攻毒浓度达3.5×109 CFU/mL时小鼠死亡率达100%,且病理切片结果表明,Sa LZ1菌株能引起小鼠内脏组织发生病变。以上结果为西藏林芝市藏猪葡萄球菌的防控提供理论依据。

关键词: 藏猪; 检菌率; 产色葡萄球菌; 分离鉴定; 生物学特性

Abstract: In order to understand the infection of Staphylococcus disease in healthy Tibetan pigs in Linzhi city,Tibet.232 samples were collected from the slaughterhouse in Bahe town,Gongbujiangda county,Linzhi city,Tibet,the teaching practice ranch at the Tibet Agriculture and Animal Husbandry University in Bayi district,Linzhi city,and the slaughterhouse in Bayi district,Linzhi city.These samples were isolated,cultured and identified,and one isolated strain was systematically studied for biological characteristics.Epidemiological survey results showed that 73 of 232 samples were positive for Staphylococcus,the positive rate was 31.47%,16 positive samples were detected in the slaughterhouse of Bahe town,Gongbujiangda county,Linzhi,Tibet,with a detection rate of 23.89%(16/67),43 positive samples were detected in the pastures of the teaching practice ranch at the Tibet Agriculture and Animal Husbandry University in Bayi district,Linzhi city,with a detection rate of 34.40% (43/125),14 positive samples were detected in the slanghterhouse in Bayi district,Linzhi city,with a detection rate of 35.00% (14/40).Through PCR identification,gene sequencing and biochemical identification,5 strains of Staphylococcus chromogenes were isolated,and one of them (Sa LZ1) was studied systematically.The biological characteristics of the isolated strains showed that the Sa LZ1 strain formed round white convex colonies on the agar medium,with regular edges,smooth surfaces,and opaque white colonies.The isolates were Gram-positive cocci bacteria.PCR sequencing results showed that the isolate had the highest homology with Staphylococcus chromogenes (GenBank accession No.:MG576253.1),positive for serum inulin,manicol,glucose,saccharose,lactose,negative for arabinose,sodium hippurate,raffinose,xylose,urea,etc,consistent with the biochemical characteristics of Staphylococcus chromogenes.The logarithmic growth period of isolated bacteria was 5 to 10 h.The pathogenicity test of mice showed that the mortality rate of mice reached 100% when the concentration of Sa LZ1 strain reached 3.5×109 CFU/mL.And the results of pathological section showed that Sa LZ1 strain could cause the visceral tissue of mice to develop pathological changes.The above results provided a theoretical basis for the prevention and control of Staphylococcus suis in Linzhi,Tibet.

Key words: Tibetan pig; detection rate; Staphylococcus chromogenes; isolation and identification; biological characteristics

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