痤疮丙酸杆菌的分离鉴定及其对瘤胃微生物发酵的影响  

Isolation,identification and rumen fermentation characteristics of Propionibacterium acnes

作　　者：吴凌[1] 赵明娟[1] 夏成[1] 倪宏波[1] 张洪友[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319

出　　处：《微生物学报》2009年第2期168-173,共6页Acta Microbiologica Sinica

基　　金：黑龙江省科技厅攻关项目(GC05B504)~~

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。[Objectives]Characteristic of energy metabolism in ruminant is a negative energy balance in perinatal period. Propionic acid from ruminal microbe fermentation is a vital glyconeogenesis substrate for preventing negative energy balance. We isolated and screened a Propionibacterium acnes strain from health cow rumen fluid, and studied its rumen fermentation characteristics. [Methods] A Propionibacterium acnes strain from rumen fluid of health cow with permanent rumen fistula under sterile condition was isolated by segregation procedure of anaerobic bacterium and Sodium Lactate Broth （SLB）, and identified by extraction of the genome DNA, cloning of the 16S rRNA gene, and sequencing. We studied the effect of the strain on pH, volatile fatty acid and lactic acid in rumen fluid in vitro and in vivo. [Results] A bacterium isolated from health cow rumen fluid was identified as Propionibacterium acnes by morphology, biochemical characteristics and sequence homology. In vitro, pH in rumen fluid decreased to the lowest after rumen fermentation of the strain for 12 h, then increased gradually. However, concentration of volatile fatty acid, such as acetic acid, propionic acid and butyric acid, increased to the highest after rumen fermentation of the strain for 12 h, then decreased gradually in vitro. The concentration of lactic acid and ratio of acetate to propionate decreased overall in vitro. In vivo, pH in rumen fluid decreased overall, concentration of the volatile fatty acid increased overall. [Conclusions] A strain of Propionibacterium acnes was isolated successfully from health cow rumen fluid. It is an important basis to develop microecological preparation for preventing cows＇ negative energy balance in perinatal period in future.

关 键 词：痤疮丙酸杆菌 分离鉴定 瘤胃微生物发酵 

分 类 号：S858.23[农业科学—临床兽医学]