大熊猫肠道芽孢杆菌的分离鉴定及部分生物学特性  

Isolation, identification and partial biology research of Bacillus in giant panda intestines

作　　者：李进[1] 钟志军[1] 苏怀益 周紫峣[1] 李魏[1] 刘福芮 兰景超 周潇潇 张文平 彭广能[1] 

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]成都大熊猫繁育研究基地,四川成都610081 [3]成都市动物疫病预防控制中心,四川成都610000

出　　处：《微生物学通报》2016年第2期351-359,共9页Microbiology China

基　　金：国家自然科学基金面上项目(No.31272620);成都大熊猫繁育研究基金会项目(No.002Z3102)~~

摘　　要：【目的】分析大熊猫肠道中芽孢杆菌的种类、纤维素分解能力、抗微生物作用和常用抗生素药物敏感性。【方法】利用芽孢耐高温特性分离菌株,基于16S r RNA基因序列构建系统发育树,通过测量芽孢杆菌在刚果红纤维素培养基上的分解圈分析其纤维素分解能力,采用牛津杯法测定芽孢杆菌的抑菌能力,结合软件分析抑菌能力和进化树之间的关系,通过PCR调查芽孢杆菌的抗菌肽分布规律,最后通过药敏试验检测芽孢杆菌是否对常用抗生素敏感。【结果】共分离得到21株芽孢杆菌;进化树显示,这些芽孢杆菌分为6个类别(Category);羧甲基纤维素钠水解结果显示,所有菌株均能分解纤维素;大部分芽孢杆菌菌株对3种肠道病原菌有较强的抑制能力,聚类分析表明,菌株的抗菌能力与基于16S r RNA基因的分类有一定的关联性;66.67%(14/21)的菌株中可以检测到2个或3个抗菌肽基因;药敏试验结果显示,菌株整体药物耐受率低,仅为7.54%(19/264),但仍有少数菌株对抗生素耐受。【结论】分离菌株种类丰富,分布平均,且均具有纤维素分解能力。21株菌株都含有抗菌肽基因,代谢产物对3种肠道病原菌具有明显抑制作用。常用抗生素耐受性低,对规范临床用药具有指导性。[Objective] The purpose of this research was to analyze the Bacillus species, cellulose decomposition, antimicrobial activity and antibiotic resistance in giant panda intestines. [Methods] Bacillus was isolated based on the resistance of high temperature, then to construct the phylogenetic tree with 16 S r RNA gene sequences. Hydrolyzed circle on the cellulose-congo red medium was measured to detect the cellulose decomposition. The oxford cup method was adopted to examine the bacteriostatic ability and then we analysed the relationship between the bacteriostatic ability and phylogenetic tree by software. The distribution regularity of Bacillus antibacterial peptides was detected by PCR. At last, susceptibility testing was put into use to detect the antibiotic resistance of Bacillus. [Results] Twenty-one strains was isolated and divided into 6 categories. All of the strains are able to decompose the cellulose on different levels by measuring the hydrolysis circle diameter. Most strains can inhibit the enteric pathogens obviously. Cluster analysis shows that the antibacterial ability is relevant to gene classification based on 16 S r RNA gene in some cases. Susceptibility testing shows that most Bacillus strains are sensitive to common antibiotics, but a few strains are still resistant to the antibiotics. [Conclusion] The Bacillus strains isolated in giant panda intestines are abundant and able to decompose cellulose. The metabolic product of the antimicrobial peptide gene contained in all of 21 Bacillus strains can significantly inhibit three kinds of enteric pathogens. The Bacillus are sensitive to common antibiotics which is helpful to standardize clinical medicine.

关 键 词：大熊猫 芽孢杆菌 16S RRNA基因 耐药性 肠道菌群 纤维素分解 

分 类 号：S865.3[农业科学—野生动物驯养] S852.6[农业科学—畜牧兽医]