牛瘤胃微生物的分离鉴定与生化特性分析

doi:10.16431/j.cnki.1671-7236.2022.05.017

摘要/Abstract

摘要： 【目的】通过对瘤胃液中分离所得的菌株进行研究，为益生菌制剂的制备提供基础数据。【方法】采集10头健康荷斯坦奶牛的瘤胃液，通过涂布、特性培养、纯化等步骤获得单一菌株，提取细菌DNA，进行16S rDNA的PCR扩增。通过16S rDNA基因测序鉴定后进行序列比对并构建系统发育树，确定菌株种类。对不同种类的细菌进行0~48 h的培养测定其生长特性，并进行耐酸碱和耐胆盐试验。【结果】通过涂布分离菌株得到20株分离菌，经PCR扩增后测序发现20株菌属于7种不同的菌，大致确定L7为解淀粉芽孢杆菌、K7为枯草芽孢杆菌、S7为嗜麦芽窄食单胞菌、C2为地衣芽孢杆菌、M7为马链球菌、F7为弗氏酸柠檬杆菌、T7为吉氏库特菌。通过生长曲线可以看出，7株菌分别在24~36 h达到最快生长期。M7和K7对酸有较强的耐受性，而S7、C2和F7对酸性较为敏感，C2对碱的耐受性最强，而T7对胆盐的耐受性最强，S7对胆盐最不耐受。【结论】本研究从瘤胃中分离得到的7个菌株对酸碱和胆盐具有不同程度的耐受性，以及最适生长周期。

关键词: 荷斯坦奶牛; 瘤胃微生物; 生化特性; 生长曲线

Abstract: 【Objective】 In this experiment,the strains isolated from the rumen juice were studied to provide basic data for the preparation of probiotic preparations.【Method】 The rumen juice of 10 healthy Holstein dairy cattle was collected,and a single strain was obtained by coating,characteristic culture and purification.The bacterial DNA was extracted and amplified by 16S rDNA PCR.After 16S rDNA gene sequencing and identification,sequence alignment was carried out and evolutionary tree was constructed to determine the species of strains.Different kinds of bacteria were cultured for 0-48 hours to determine their growth characteristics,and acid,alkali and bile salt tolerance experiments were carried out to determine their different tolerance.【Result】 20 strains were obtained by coating the isolated strains.After PCR amplification and sequencing,it was found that 20 strains belonged to 7 different bacteria.It was roughly determined that L7 was Bacillus amyloliquefaciens,K7 was Bacillus subtilis,S7 was Stenotrophomonas maltophilia,C2 was Bacillus licheniformis,M7 was Streptococcus equi,F7 was Citrobacter freundii,and T7 was Kurt gigasii.From the growth curve,it could be seen that the optimum growth time of the seven strains reached the fastest growth period in 24-36 h respectively.M7 and K7 had strong tolerance to acid,while S7,C2 and F7 were more sensitive to acid,C2 had the strongest tolerance to alkali,T7 had the strongest tolerance to bile salt,and S7 had the least tolerance to bile salt.【Conclusion】 The seven strains isolated from rumen in this study had different degrees of tolerance to acid-base and bile salt,and the optimal growth cycle.

Key words: Holstein dairy cattle; rumen microorganisms; biochemical characteristics; growth curve

中图分类号:

S811.6

李畅洋, 高炳南, 任雨龙, 张昊, 孙瑜, 魏笑, 王秋菊. 牛瘤胃微生物的分离鉴定与生化特性分析[J]. 中国畜牧兽医, 2022, 49(5): 1757-1764.

LI Changyang, GAO Bingnan, REN Yulong, ZHANG Hao, SUN Yu, WEI Xiao, WANG Qiuju. Isolation,Identification and Biochemical Characteristics Analysis of Bovine Rumen Functional Bacteria[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1757-1764.

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