1株牦牛瘤胃源无色杆菌的鉴定及生物学特性评价

doi:10.16431/j.cnki.1671-7236.2022.12.038

摘要/Abstract

摘要： 【目的】对分离得到的1株牦牛瘤胃源菌株进行鉴定和生物学特性分析，开发新型动物源益生菌。【方法】通过显微镜和菌落观察试验菌株的形态学特征，采用VITEK2微生物生化自动鉴定仪鉴定其生化特性，并对其16S rDNA进行PCR扩增、相似性比对和遗传进化树构建。进一步对其生长性能、产酸能力、自凝聚、疏水性、溶血性和耐药性、耐酸耐胆盐及胃肠环境适应性等生物学特性进行研究。【结果】试验菌株经过形态学、分子鉴定和16S rDNA扩增鉴定为无色杆菌（Achromobacter），命名为无色杆菌YKS2。该菌株2 h进入对数生长期，持续增长至20 h后开始出现平台期并缓慢下降。发酵48 h后pH未降低，说明该菌株不具产酸性能；自凝聚力为28.7%，表面疏水性在氯仿中达到77.1%，其他溶剂均>60.0%，平均达到60%以上，表明细菌有一定的黏附能力；在血琼脂平板培养48 h后未出现溶血环；通过K-B法测试了对14种抗生素的敏感性，发现其对常见的12种抗生素敏感；在pH 2.0的培养基中不能存活，在pH 4.0的培养基中存活率为61.4%；对0.1%和0.3%胆盐培养基有较高的耐受；在人工胃液重悬后培养3 h的细菌存活率为78.18%，在人工肠液中处理4、8 h后，与对照组相比，菌落数极显著增长（P<0.05），说明YKS2菌株能够耐受一定程度的人工肠液。【结论】牦牛瘤胃源YKS2菌株为肠杆菌科无色杆菌亚种，无溶血、产酸及耐强酸特性，非耐药菌株，增殖性能较好，具有耐胆盐、耐胃肠液、自凝聚及表面疏水特性，可作为一种候选益生菌菌株进行进一步的评价和研究。

关键词: 牦牛; 瘤胃; 无色杆菌; 生物学特性

Abstract: 【Objective】 In order to develop a new type of animal-derived probiotics, a yak rumen strain was identified and its biological characteristics were analyzed.【Method】 The tested strains were identified by microscope and colony observation, and its biochemical characteristics was identified by VITEK2 microbiological automatic identification instrument.The 16S rDNA was amplified by PCR, and similarity comparison and genetic evolution tree construction were carried out.The growth performance, acid producing ability, self-coagulation, hydrophobicity, hemolysis and drug resistance, acid and bile salt resistance and gastrointestinal environmental adaptability were studied.【Result】 After morphological, molecular identification and 16S rDNA amplification, the test strain was identified as Achromobacter and named as Achromobacter YKS2.The strain entered logarithmic phase at 2 h, and continued to grow to 20 h, and began to plateau and slowly decline.The pH did not decrease after 48 h fermentation, indicating that the strain did not have acid production ability.The self-cohesion was 28.7%, the surface hydrophobicity was 77.1% in chloroform, and the other solvents were more than 60.0%, with an average of more than 60%, indicating that the bacteria had certain adhesion ability.There was no hemolytic ring after 48 h incubation on blood agar plate.The sensitivity to 14 antibiotics was tested by K-B method, and it was found that it was sensitive to 12 common antibiotics.The strain could not survive in the medium with pH 2.0, and the survival rate was 61.4% in the medium with pH 4.0.It had high tolerance to bile salt medium containing 0.1% and 0.3%.The survival rate of bacteria cultured in artificial gastric juice for 3 h was 78.18%.After treatment in artificial intestinal juice for 4 and 8 h, the number of colonies increased significantly compared with the control group (P<0.05), indicating that YKS2 strain could tolerate artificial intestinal juice to a certain extent.【Conclusion】 YKS2 strain derived from yak rumen was a subspecies of Achromobacillus of Enterobacteriaceae with no hemolysis, acid production and strong acid resistance.It was a non-drug resistant strain with good proliferation performance, and had the properties of bile salt resistance, gastric fluid resistance, self-condensation and surface hydrophobicity.It could be used as a candidate probiotic strain for further evaluation and study.

Key words: yak; rumen; Achromobacter; biological characteristics

中图分类号:

S852.69

李大鹏, 曹春华, 付鹏程, 张景艳, 张康, 王磊, 张凯, 刘琴, 包永占, 李建喜. 1株牦牛瘤胃源无色杆菌的鉴定及生物学特性评价[J]. 中国畜牧兽医, 2022, 49(12): 4909-4919.

LI Dapeng, CAO Chunhua, FU Pengcheng, ZHANG Jingyan, ZHANG Kang, WANG Lei, ZHANG Kai, LIU Qin, BAO Yongzhan, LI Jianxi. Identification and Biological Characteristics Evaluation of an Achromobacter Strain Isolated from Rumen of Yak[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4909-4919.

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