痤疮丙酸杆菌的分离鉴定及其对瘤胃微生物发酵的影响
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黑龙江省科技厅攻关项目(GC05B504)


Isolation and identification of Propionibacterium Acnes and its effect on the fermentation of rumen microbes
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Supported by Heilongjiang province science and technology project(GC05B504)

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    摘要:【目的】奶牛围产期能量代谢的特点是能量负平衡,瘤胃发酵产生的丙酸是奶牛糖异生供能的主要底物,对预防奶牛能量负平衡具有重要的意义。本研究旨在从健康奶牛瘤胃液中分离、筛选出以产丙酸为主的痤疮丙酸杆菌,研究其瘤胃发酵特性。【方法】无菌采取装有瘤胃瘘奶牛的瘤胃液,按照厌氧菌分离步骤,通过丙酸生成菌株的特异性培养基SLB进行筛选,提取分离菌的基因组DNA,克隆其16S rRNA基因,进行序列测定,分离出一株痤疮丙酸杆菌。通过体内外发酵试验研究痤疮丙酸杆菌对瘤胃液pH、挥发性脂肪酸和乳酸的影响。【结果】通过形态学观察、生化反应和序列分析证实所分离的一株产丙酸的杆菌为痤疮丙酸杆菌。该菌株在体外发酵过程中,瘤胃液pH先下降,在12h时降至最低,随后上升;乙酸、丙酸、丁酸等挥发性脂肪酸先升高,于12h时升至最高,随后又降低;乳酸浓度和乙酸/丙酸总体上一直下降;在体内发酵过程中,pH总体上下降;乙酸、丙酸、丁酸等挥发性脂肪酸总体上升。【结论】在国内首次从健康牛瘤胃液中成功分离出一株痤疮丙酸杆菌,为今后研发预防奶牛能量负平衡的微生态制剂奠定基础。

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    Abstract: [Objectives] Characteristic of energy metabolism in ruminant is negative energy balance in perinatal period. Propionic acid from ruminal microbes fermentation is a vital glyconeogenesis substrate for prevention of negative energy balance. We intend to isolate and screen a strain of Propionibacterium acnes from health cow rumen fluid, and study its rumen fermentation characteristics. [Methods] A strain of Propionibacterium acnes from rumen fluid of health cow with permanent rumen fistula under sterile condition was isolated by segregation procedure of anaerobic bacterium and specific medium SLB, and identified by extraction of the genome DNA of isolation bacterium, clone of the 16S rRNA gene, and sequencing of the strain. We study effect of the stain on pH, volatile fatty acid and lactic acid in rumen fluid by vitro and vivo test. [Results] The results showed that a strain of bacterium isolated from health cow rumen fluid was identified as the Propionibacterium acnes by morphology, biochenmical characteristics, sequence homology. In vitro test of the strain, pH in rumen fluid first dropped, to the lowest at 12 h, then increased gradually; concentration of volatile fatty acid, including acetic acid, propionic acid and butyric acid, first increased, to the highest at 12 h, then decreased gradually; the lactic acid concentration and ratio of acetate to propionate ratio decreased overall. In vivo test of the strain, pH in rumen fluid decrease overall; concentration of the volatile fatty acid increased overall. [Conclusions] A strain of Propionibacterium acnes was first isolated successfully from health cow rumen fluid at home, it is an important basis for us to develop microecological preparation for prevention of cows’ negative energy balance in perinatal period in future.

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吴凌,赵明娟,夏成,倪宏波,张洪友. 痤疮丙酸杆菌的分离鉴定及其对瘤胃微生物发酵的影响. 微生物学报, 2009, 49(2): 168-173

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  • 收稿日期:2008-07-01
  • 最后修改日期:2008-10-28
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