1株麂子源麦芽香肉杆菌的分离鉴定及生物学特性分析

doi:10.16431/j.cnki.1671-7236.2023.01.040

摘要/Abstract

摘要： 【目的】探究昆明轿子雪山自然保护区死亡麂子体内潜在微生物的生物学特性，以揭示其与该麂子死亡是否存在相关性，同时为昆明轿子雪山自然保护区的微生物多样性提供数据支持。【方法】无菌采集死亡麂子的肩部肌肉组织，对其进行细菌分离培养，采用革兰氏染色镜检、生化试验及16S rRNA基因扩增鉴定后进行序列比对并构建系统发育树，确定分离菌株种类并对分离菌株进行药敏试验和动物回归试验。【结果】分离菌株在胰酪大豆胨琼脂培养基（TSA）和血琼脂培养基上菌落形态为圆形、边缘整齐、表面光滑，呈乳白色；在麦康凯培养基（MAC）上不生长，革兰氏染色镜检见紫色杆菌。生化试验结果显示，分离菌株不能发酵蔗糖、棉子糖、山梨醇等；苯丙氨酸、鸟氨酸、赖氨酸、V-P试验、葡萄糖产气阳性。16S rRNA基因序列和系统发育树分析发现，该菌株与GenBank数据库中登录的麦芽香肉杆菌菌株MMF-23（GQ304931.1）相似性达100%。结合形态学、生化特征等，鉴定分离菌株为麦芽香肉杆菌，命名为BJT86，并将16S rRNA序列提交GenBank，获得登录号为ON966116.1。药敏试验结果显示，分离菌株对青霉素、阿莫西林、哌拉西林等6种抗菌药表现为耐药，对头孢唑啉、头孢噻肟、头孢舒巴坦等18种抗菌药敏感。动物回归试验表明分离菌株无致病性。【结论】本试验从昆明轿子雪山自然保护区死亡麂子肩部肌肉组织首次成功分离得到1株麦芽香肉杆菌，可为食品安全提供新的参考方向，同时为益生菌制剂的制备提供基础数据。

关键词: 麦芽香肉杆菌; 分离鉴定; 药敏试验; 回归试验

Abstract: 【Objective】 The purpose of the experiment was to explore the biological characteristics of potential microorganisms in the dead barking deer in the Jiaozi Snow Mountain Nature Reserve, Kunming, reveal whether there was a correlation between them and the death of the barking deer, and to provide data support for microbial diversity in Kunming Jiaozi Snow Mountain Nature Reserve.【Method】 Part of the muscle tissue of the dead barking deer was collected aseptically in the experiment.The bacteria were isolated and cultured.After identification by Gram staining microscopy, biochemical test and 16S rRNA gene amplification, sequence comparison was conducted and phylogenetic tree was constructed to determine the type of the isolated strain, and drug sensitivity test and animal regression test were conducted on the isolated strain.【Result】 The colony morphology of the isolated strain on TSA and blood agar medium was round, with neat edges, smooth surface and milky white color, it did not grow on MAC medium, Gram staining microscope showed purple bacilli.The biochemical test results showed that, the isolated strain could not ferment sucrose, raffinose, sorbitol, etc., phenylalanine, ornithine, lysine, V-P test, and glucose gas production were positive.The 16S rRNA gene sequence and phylogenetic tree analysis showed that the strain was 100% similar to the Carnobacterium maltaromaticum (C.maltobotulinum) strain MMF-23 (GQ304931.1) in the GenBank.Combined with morphological, biochemical features, the isolate was identified as C.maltaromaticum, named BJT86, and the 16S rRNA sequence was submitted to GenBank to obtain the accession number ON966116.1.The results of drug sensitivity test showed that the isolated strain was resistant to 6 kinds of antibiotics such as penicillin, amoxicillin and piperacillin, and sensitive to 18 kinds of antibiotics such as cefazolin, cefotaxime and cefsulbactam.Animal regression test showed that the isolated strain was not pathogenic.【Conclusion】 In this experiment, a strain of C.maltaromaticum was successfully isolated for the first time from the muscle tissue of the dead barking deer in the Jiaozi Snow Mountain Nature Reserve in Kunming, which could provide a new reference direction for food safety and basic data for the preparation of probiotics.

Key words: Carnobacterium maltaromaticum; isolation and identification; drug susceptibility test; regression test

中图分类号:

Q939.11

杨福梅, 杨林富, 曾成, 杨志, 王康, 段纲, 代飞燕. 1株麂子源麦芽香肉杆菌的分离鉴定及生物学特性分析[J]. 中国畜牧兽医, 2023, 50(1): 398-407.

YANG Fumei, YANG Linfu, ZENG Cheng, YANG Zhi, WANG Kang, DUAN Gang, DAI Feiyan. Isolation,Identification and Biological Characterization Analysis of a Strain of Carnobacterium maltaromaticum from Barking Deer[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 398-407.

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