藏羊源链球菌的分离鉴定及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2023.09.038

摘要/Abstract

摘要： 【目的】2022年12月西藏拉萨市某县藏羊陆续出现不明原因死亡病例,试验旨在确定引起西藏拉萨藏羊死亡的病原,研究其生物学特性,为今后对该病的防控提供科学依据。【方法】从病死藏羊病料中分离培养、纯化病原菌,通过形态特征、生化特性、16S rRNA PCR扩增及遗传进化分析对分离菌株进行鉴定;通过动物致病性试验、药物敏感性试验对分离菌株的致病性、耐药性进行研究。【结果】从病死藏羊病料组织中分离到8株革兰阳性球菌,分别命名为:Tibet-1、Tibet-2、Tibet-3、Tibet-4、Tibet-5、Tibet-6、Tibet-7、Tibet-8。生化鉴定结果显示,分离菌株均能发酵麦芽糖、乳糖、葡萄糖、山梨醇、七叶苷及水杨苷,能分解甘露醇,不能分解尿素酶、淀粉、木糖,V-P试验为阴性,结果符合链球菌的生化特性。16S rRNA基因测序结果显示,8株分离菌株与链球菌新亚种羊属链球菌亚种(Streptococcus ovis)标准株CCUG 39485T(Y17358)的相似性为95.8%~97.4%。系统进化树显示,分离菌株与羊属链球菌亚种处于同一支,而与链球菌其他种属处于不同分支。动物致病性试验表明,分离菌株对小鼠具有不同强度的致病性,死亡小鼠肝脏、肺脏等内脏器官有明显病变。药物敏感性试验结果显示,分离菌株对氯霉素、氧氟沙星、环丙沙星敏感,对头孢拉定、万古霉素、克林霉素等13种抗菌药存在不同程度的耐药性,其中7株分离菌株存在多重耐药性。【结论】本研究成功分离到8株藏羊源致病性羊属链球菌,不同分离菌株耐药性存在一定差异,可为临床用药和防控提供参考依据。

关键词: 藏羊; 链球菌; 分离鉴定; 生物学特性

Abstract: 【Objective】 In December 2022,unexplained death cases of Tibetan sheep were successively observed in a county of Lhasa,Tibet.This study was aimed to identify the pathogen causing the death of Tibetan sheep in Lhasa,Tibet,and study its biological characteristics,so as to provide scientific basis for the prevention and control of the disease in the future.【Method】 The pathogenic bacteria were isolated and purified from the diseased material in dead Tibetan sheep,and the isolates were identified by morphological characteristics,biochemical properties,16S rRNA PCR amplification and genetic evolutionary analysis.The pathogenicity and drug resistance of the isolates were studied by animal pathogenicity and drug susceptibility tests.【Result】 Eight Gram-positive cocci,named Tibet-1,Tibet-2,Tibet-3,Tibet-4,Tibet-5,Tibet-6,Tibet-7 and Tibet-8,were isolated from the diseased tissues in dead Tibetan sheep.Biochemical identification results showed that all the isolates could ferment maltose,lactose,glucose,sorbitol,aescin,salicylin and decomposed mannitol,and couldn’t decompose urease,starch and xylose.V-P test was negative,and the results conformed to the biochemical characteristics of Streptococcus.The results of 16S rRNA gene sequencing showed that the eight isolates were 95.8%-97.4% similarity to the standard strain of Streptococcus ovis subspecies CCUG 39485T (Y17358).The evolutionary tree showed that the isolates were in the same family as Streptococcus ovis subspecies and in a different branch from other Streptococcus species.Animal pathogenicity tests showed that the isolates were pathogenic to mice of different intensities,with significant lesions in liver,lung and other internal organs of dead mice.Drug susceptibility tests showed that the isolates were sensitive to chloramphenicol,ofloxacin and ciprofloxacin,and were resistant to 13 antimicrobials including cefradin,vancomycin and clindamycin to different degrees,and 7 of the isolates were multi-drug resistant.【Conclusion】 In this study,8 strains of Streptococcus ovis of Tibetan sheep origin were successfully isolated,and the drug resistance of different isolates differed to some extent,providing a reference for clinical medication and prevention and control.

Key words: Tibetan sheep; Streptococcus; isolation and identification; biological characteristics

中图分类号:

S852.61⁺1

王冬经, 苏中华, 元振杰, 马弘财, 曾江勇. 藏羊源链球菌的分离鉴定及生物学特性研究[J]. 中国畜牧兽医, 2023, 50(9): 3823-3832.

WANG Dongjing, SU Zhonghua, YUAN Zhenjie, MA Hongcai, ZENG Jiangyong. Isolation,Identification and Biological Characteristics of Streptococcus Isolated from Tibetan Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3823-3832.

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