牦牛源链球菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2023.02.032

摘要/Abstract

摘要： 【目的】了解西藏拉萨地区牦牛链球菌的感染情况、致病性及其耐药性,为牦牛源链球菌的研究提供新的数据。【方法】对55份牦牛鼻拭子进行细菌分离培养、菌落形态观察、革兰氏染色镜检、生化鉴定、16S rRNA 序列PCR扩增以鉴定分离菌的种属特性。通过人工感染试验小鼠评价分离菌的致病性,采用药敏纸片扩散法检测分离菌的耐药性。【结果】分离菌在血平板上长出灰白色、表面光滑的α溶血圆形菌落,革兰氏染色镜检呈链状排列的阳性球菌。分离菌可发酵葡萄糖、乳糖、海藻糖和山梨醇,不发酵麦芽糖和甘油,硝酸盐还原试验、靛基质试验、V-P试验和MR试验均呈阴性。经16S rRNA基因序列比对分析,确定有25株链球菌(Tibet-1~Tibet-25),检出率为45.46%(25/55),其中17株多动物链球菌,8株马链球菌。分离菌16S rRNA基因序列与NCBI上已发布的链球菌相似性在97.04%~99.77%之间,其中Tibet-1、Tibet-2、Tibet-3、Tibet-5、Tibet-9、Tibet-10、Tibet-11、Tibet-14、Tibet-15、Tibet-16、Tibet-17共11株牦牛源多动物链球菌与KM981770.1的相似性均在99%以上。分离的牦牛源链球菌可引起小鼠腹泻,对发病小鼠脏器进行增菌培养,均可分离到链球菌。药敏试验结果显示,分离菌株对多西环素、红霉素、青霉素共11种抗菌药敏感,对复方新诺明耐药。【结论】本研究分离了牦牛源多动物链球菌和牦牛源马链球菌,并对其致病性和耐药性进行相关分析,为该病的防控和治疗提供了参考依据。

关键词: 牦牛; 多动物链球菌; 马链球菌; 分离鉴定; 耐药性

Abstract: 【Objective】 This study was aimed to understand the infection,pathogenicity and drug resistance of Streptococcus from yak in Lhasa,Tibet,and provide new data for the study of Streptococcus from yak.【Method】 Bacterial isolation and culture,colony morphology observation,microscopic examination with Gram staining,biochemical identification and 16S rRNA sequence PCR amplification were performed on 55 yak nose swabs to identify the species characteristics of the isolates.The pathogenicity of the isolated bacteria was evaluated by the artificial infection test in mice.Finally,the drug resistance of the isolated bacteria was detected by disk diffusion method.【Result】 The isolated bacteria grew gray-white,smooth α-hemolytic round colonies on the blood plate,and Gram staining microscopic examination showed chain-like positive cocci. The isolated bacteria could ferment glucose, lactose, trehalose and sorbitol, but could not ferment maltose and glycerol. The nitrate reduction test, indigo substrate test, V-P test and MR test were negative. 16S rRNA gene sequence alignment analysis showed that,25 strains of Streptococcus (Tibet-1 to Tibet-25) were identified,and the detection rate was 45.46% (25/55),including 17 strains of Streptococcus pluranimalium and 8 strains of Streptococcus equi.The similarity between the 16S rRNA gene sequence of isolated bacteria and the Streptococci published on NCBI was 97.04%-99.77%.Among them,11 strains of Streptococcus pluranimalium from yaks,including Tibet-1,Tibet-2,Tibet-3,Tibet-5,Tibet-9,Tibet-10,Tibet-11,Tibet-14,Tibet15,Tibet-16 and Tibet-17,had more than 99% similarity with KM981770.1.The isolated Streptococcus from yak could cause diarrhea in mice. Streptococcus could be isolated from the organs of infected mice by enrichment culture.The results of drug sensitivity test showed that the isolated strains were highly sensitive to 11 antibiotics,including doxycycline,erythromycin,penicillin,and so on.It was highly resistant to compound sulfamethoxazole.【Conclusion】 This study isolated Streptococcus pluranimalium and Streptococcus equi from yak,and analyzed their pathogenicity and drug resistance,which provided a reference basis for the prevention,control and treatment of the disease.

Key words: yak; Streptococcus pluranimalium; Streptococcus equis; isolation and identification; drug resistance

中图分类号:

S852.61

王冰艺, 马弘财, 邹明昊, 樊世杰, 元振杰, 拉巴次仁, 董海龙, 曾江勇. 牦牛源链球菌的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2023, 50(2): 745-753.

WANG Bingyi, MA Hongcai, ZOU Minghao, FAN Shijie, YUAN Zhenjie, Labaciren, DONG Hailong, ZENG Jiangyong. Isolation,Identification and Drug Resistance Analysis of Streptococcus from Yak[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 745-753.

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