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短讯:食用干燥酒糟和可溶物的奶牛瘤胃液中酵母 DNA 的检测。

Short communication: Detection of yeast DNA in omasal digesta of dairy cows consuming dried distillers grains and solubles.

机构信息

Department of Animal Science, University of Nebraska-Lincoln, Lincoln 68583-0908, USA.

出版信息

J Dairy Sci. 2010 Dec;93(12):5926-9. doi: 10.3168/jds.2010-3302.

Abstract

Purine analysis is widely used to estimate microbial crude protein (MCP) flow, and the method assumes that all purines contained in feed are degraded in the rumen and that purines detected are of microbial origin. The objectives of our experiment were (1) to determine if DNA from yeast (Saccharomyces cerevisiae) contained in dried distillers grains and solubles (DDGS) escapes degradation in the rumen and (2) to estimate the proportion of yeast DNA compared with total bacterial DNA in omasal samples. Two ruminally fistulated Holstein dairy cows averaging 649 kg (SD = 42.0) and 126 d in milk (SD = 28.9) were fed in a crossover design during 2 periods of 21 d each. Treatments were (1) control, a total mixed ration (TMR) not containing DDGS and (2) a DDGS-based diet, a TMR in which DDGS were included at 30% of diet dry matter (DM). On d 20 and 21 at 0400 and 1600 h, omasal digesta samples were collected via a ruminal cannula, and DNA was extracted from each sample in duplicate. The DNA samples were subjected to a real-time PCR assay to detect the presence of DNA from yeast. Forward and reverse primers and a probe were designed to target a DNA segment contained on the second chromosome of Saccharomyces cerevisiae. Real-time PCR amplification curves indicated the presence of yeast DNA in samples from both treatments. Specifically, the estimate of relative abundance of yeast DNA from digesta samples collected from animals consuming the diet containing DDGS was 9.46 ± 0.67/g of DM and was significantly higher than that from animals consuming no DDGS, which was observed to be 0.091 ± 0.67/g of DM. Omasal samples were also analyzed for total bacterial DNA. Primers and a probe were designed from DNA encoding part of the 16S rRNA. When the DDGS-based diet was fed, the relative abundance of total bacterial DNA tended to increase from 610 to 626±3.82/g of DM. Results suggest that yeast DNA is detected in the omasum and this is increased when cows consume DDGS but it does not represent a significant proportion of total microbial DNA in the omasal digesta samples.

摘要

嘌呤分析被广泛用于估计微生物粗蛋白(MCP)的流量,该方法假设饲料中所含的所有嘌呤都在瘤胃中降解,并且检测到的嘌呤都来自微生物。我们实验的目的是(1)确定酵母(酿酒酵母)的 DNA 是否从包含在干酒糟和可溶物(DDGS)中的 DNA 逃脱瘤胃降解,(2)估计在腔样本中酵母 DNA 与总细菌 DNA 的比例。在两个周期的 21 天中,每个周期平均为 649 公斤(SD=42.0)和 126 天的哺乳期(SD=28.9)的泌乳荷斯坦奶牛通过交叉设计进行了饲喂。处理方法为(1)对照,不含 DDGS 的全混合日粮(TMR)和(2)以 DDGS 为基础的日粮,DDGS 占日粮干物质(DM)的 30%。在 0400 和 1600 h 时,在第 20 天和第 21 天,通过瘤胃套管收集腔消化物样本,并从每个样本中提取 DNA 进行双重复制。将 DNA 样本进行实时 PCR 检测,以检测酵母 DNA 的存在。设计了正向和反向引物以及探针,以靶向酿酒酵母第二染色体上的 DNA 片段。实时 PCR 扩增曲线表明,来自两种处理方法的消化物样本中均存在酵母 DNA。具体来说,从食用含有 DDGS 的日粮的动物的消化物样本中检测到的酵母 DNA 的相对丰度为 9.46 ± 0.67/g DM,明显高于未食用 DDGS 的动物的 0.091 ± 0.67/g DM。还对腔样本中的总细菌 DNA 进行了分析。设计了用于部分 16S rRNA 编码的引物和探针。当喂食基于 DDGS 的饮食时,总细菌 DNA 的相对丰度从 610 增加到 626±3.82/g DM。结果表明,在腔中检测到酵母 DNA,当奶牛食用 DDGS 时,这种情况会增加,但它并不代表腔消化物样本中总微生物 DNA 的重要比例。

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