Agriculture and Agri-Food Canada, Dairy and Swine Research and Development Centre, Sherbrooke, Quebec, Canada.
J Dairy Sci. 2013 Aug;96(8):5237-48. doi: 10.3168/jds.2012-6481. Epub 2013 Jun 13.
Methane produced by the methanogenic Archaea that inhabit the rumen is a potent greenhouse gas and represents an energy loss for the animal. Although several strategies have been proposed to mitigate enteric CH4 production, little is known about the effects of dietary changes on the microbial consortia involved in ruminal methanogenesis. Thus, the current study aimed to examine how the metabolically active microbes are affected when dairy cows were fed diets with increasing proportions of corn silage (CS). For this purpose, 9 ruminally cannulated lactating dairy cows were used in a replicated 3 × 3 Latin square design and fed a total mixed ration (60:40 forage:concentrate ratio on a dry matter basis) with the forage portion being either alfalfa silage (0% CS), corn silage (100% CS), or a 50:50 mixture (50% CS). Enteric CH4 production was determined using respiration chambers and total rumen content was sampled for the determination of fermentation characteristics and molecular biology analyses (cDNA-based length heterogeneity PCR, quantitative PCR). The cDNA-based length heterogeneity PCR targeting active microbes revealed similar bacterial communities in cows fed 0% CS and 50% CS diets, whereas important differences were observed between 0% CS and 100% CS diets, including a reduction in the bacterial richness and diversity in cows fed 100% CS diet. As revealed by quantitative PCR, feeding the 100% CS diet increased the number of total bacteria, Prevotella spp., Archaea, and methanogenic activity, though it reduced protozoal number. Meanwhile, increasing the CS proportion in the diet increased propionate concentration but decreased ruminal pH, CH4 production (L/kg of dry matter intake), and concentrations of acetate and butyrate. Based on these microbial and fermentation changes, and because CH4 production was reduced by feeding 100% CS diet, this study shows that the use of cDNA-based quantitative PCR to estimate archaeal growth and activity is not reliable enough to reflect changes in ruminal methanogenesis. A more robust technique to characterize changes in archaeal community structures will help to better understand the microbial process involved in ruminal methanogenesis and, hence, enabling the development of more effective dietary CH4 mitigation strategies.
瘤胃中栖息的产甲烷古菌产生的甲烷是一种强效温室气体,代表着动物的能量损失。尽管已经提出了几种策略来减轻肠道 CH4 的产生,但对于饮食变化对瘤胃甲烷生成中涉及的微生物群落的影响知之甚少。因此,本研究旨在研究当奶牛饲喂含有不同比例玉米青贮(CS)的日粮时,代谢活跃的微生物会受到怎样的影响。为此,本研究使用 9 头瘤胃内置有套管的泌乳奶牛,采用 3×3 拉丁方设计进行了重复性试验,并以总混合日粮(基于干物质的 60:40 粗饲料:精料比)进行饲养,其中粗饲料部分分别为紫花苜蓿青贮(0%CS)、玉米青贮(100%CS)或 50:50 混合物(50%CS)。使用呼吸室来确定肠道 CH4 的产生,并对总瘤胃液进行采样以确定发酵特性和分子生物学分析(基于 cDNA 的长度异质性 PCR、定量 PCR)。基于 cDNA 的长度异质性 PCR 靶向活性微生物的方法表明,饲喂 0%CS 和 50%CS 日粮的奶牛具有相似的细菌群落,而 0%CS 和 100%CS 日粮之间存在重要差异,包括饲喂 100%CS 日粮的奶牛的细菌丰富度和多样性降低。定量 PCR 显示,饲喂 100%CS 日粮会增加总细菌、普雷沃氏菌属、古菌和产甲烷活性的数量,尽管它会降低原生动物的数量。同时,日粮中 CS 比例的增加会增加丙酸浓度,但会降低瘤胃 pH 值、CH4 产生量(L/干物质采食量)以及乙酸和丁酸的浓度。基于这些微生物和发酵变化,并且由于饲喂 100%CS 日粮会减少 CH4 的产生,本研究表明,基于 cDNA 的定量 PCR 估计古菌生长和活性的方法不够可靠,无法反映瘤胃甲烷生成的变化。一种更可靠的技术来描述古菌群落结构的变化将有助于更好地理解瘤胃甲烷生成过程中的微生物过程,从而能够开发更有效的日粮 CH4 减排策略。