Universität Hohenheim, Institut für Tierernährung, Emil-Wolff-Strasse 10, Stuttgart, Germany.
Anaerobe. 2010 Aug;16(4):412-9. doi: 10.1016/j.anaerobe.2010.05.002. Epub 2010 May 21.
This study examined whether different corn silage to grass silage ratios in ruminant rations and different grinding levels of the feed affect the composition of the ruminal Bacteroides-Prevotella community in vitro. Three diets, composed of 10% soybean meal as well as of different corn silage and grass silage proportions, were ground through 1mm or 4mm screened sieves and incubated in a semi-continuous rumen simulation system. On day 14 of the incubation microbes were harvested by centrifugation from the liquid effluent of fermenter vessels. Microbial DNA was extracted for single strand conformation polymorphism (SSCP) analysis of 16S rRNA genes followed by sequencing of single SSCP bands. Fluorescence in situ hybridization (FISH) and real-time quantitative (q) PCR were used to quantify differences in the relative abundance of Bacteroides-Prevotella and Prevotella bryantii. SSCP profiles revealed a significant influence of the forage source as well as of the feed particle size on the community structure of the Bacteroides-Prevotella group. Different, phylogenetically distinct, so far uncultured Prevotella species were detected by sequence analysis of several treatment-dependent occurring SSCP bands indicating different nutritional requirements of these organisms for growth. No quantitative differences in the occurrence of Bacteroides-Prevotella-related species were detected between diets by FISH with probe BAC303. However, real-time qPCR data revealed a higher abundance of P. bryantii with increasing grass silage to corn silage ratio, thus again indicating changes within the community composition of the Bacteroides-Prevotella group. As P. bryantii possesses high proteolytic activity its higher abundance may have been caused by the higher contents of crude protein in the grass silage containing diets. To conclude, results of this study show an influence of the forage source on the ruminal community of Bacteroides-Prevotella. Furthermore, they suggest an effect of the feed particle size on this bacterial group.
本研究旨在探讨反刍动物日粮中不同的玉米青贮与牧草青贮比例以及不同的饲料粉碎水平对体外瘤胃拟杆菌-普雷沃氏菌群落组成的影响。三种日粮由 10%的豆粕以及不同比例的玉米青贮和牧草青贮组成,分别通过 1mm 或 4mm 筛网粉碎,并在半连续瘤胃模拟系统中进行培养。在培养的第 14 天,通过离心从发酵罐液体流出物中收获微生物。提取微生物 DNA 进行 16S rRNA 基因的单链构象多态性(SSCP)分析,然后对单 SSCP 带进行测序。荧光原位杂交(FISH)和实时定量(q)PCR 用于定量拟杆菌-普雷沃氏菌和普雷沃氏菌属Bryantii 的相对丰度差异。SSCP 图谱显示,饲草料来源以及饲料颗粒大小对拟杆菌-普雷沃氏菌群的群落结构有显著影响。通过对几种处理相关出现的 SSCP 带的序列分析,检测到不同的、系统发育上不同的、迄今为止尚未培养的普雷沃氏菌属,表明这些生物体对生长有不同的营养需求。通过探针 BAC303 的 FISH 检测,未发现不同日粮之间 Bacteroides-Prevotella 相关物种的出现存在定量差异。然而,实时 qPCR 数据显示,随着牧草青贮与玉米青贮比例的增加,普雷沃氏菌属Bryantii 的丰度增加,这再次表明拟杆菌-普雷沃氏菌群的群落组成发生了变化。由于普雷沃氏菌属Bryantii 具有较高的蛋白水解活性,其丰度的增加可能是由于含牧草青贮日粮中粗蛋白含量较高所致。综上所述,本研究结果表明饲草料来源对瘤胃拟杆菌-普雷沃氏菌群落有影响。此外,它们还表明饲料颗粒大小对该细菌群有影响。
FEMS Microbiol Lett. 2010-1-27