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持续或短期高谷物喂养后,奶牛瘤胃上皮的短链脂肪酸吸收、基因表达和细菌群落的适应性反应。

Adaptive responses in short-chain fatty acid absorption, gene expression, and bacterial community of the bovine rumen epithelium recovered from a continuous or transient high-grain feeding.

机构信息

Institute of Animal Nutrition and Functional Plant Compounds, Veterinaerplatz 1, 1210 Vienna, Austria; Animal Gut Health Research Cluster, Veterinaerplatz 1, 1210 Vienna, Austria.

Institute of Animal Nutrition and Functional Plant Compounds, Veterinaerplatz 1, 1210 Vienna, Austria; Animal Gut Health Research Cluster, Veterinaerplatz 1, 1210 Vienna, Austria; Institute for Milk Hygiene, Milk Technology and Food Science, University of Veterinary Medicine Vienna, Veterinaerplatz 1, 1210 Vienna, Austria.

出版信息

J Dairy Sci. 2019 Jun;102(6):5361-5378. doi: 10.3168/jds.2018-15691. Epub 2019 Apr 17.

Abstract

The feeding of high-grain diets to dairy cows commonly results in lowered pH and ruminal dysbiosis, characterized by changes in absorption dynamics of short-chain fatty acids (SCFA) across the reticuloruminal wall, epithelial function, and the epithelial bacteria community structure. Therefore, the present study evaluated the effect of high-grain feeding persistence on the absorption kinetics of reticuloruminal SCFA, gene expression in the rumen epithelium, and the associated shifts in the epithelial bacteria in cows recovering from either a long-term continuous high-grain feeding model or a long-term transient high-grain feeding model. In a crossover study design, 8 nonlactating Holstein cows were fed 60% concentrate either continuously for 4 wk (continuous) or with a 1-wk break in the second week of the high-grain feeding (transient). After the high-grain feeding, all animals were fed a diet of 100% forage (recovery) for an additional 8 wk. Rumen papilla biopsies and SCFA absorption measurements were taken at the start of the trial (baseline), after the 4-wk high-grain feeding (49 d), after 2-wk recovery forage feeding (63 d), and after 8-wk recovery forage (105 d). Absorption of SCFA was determined in vivo using the washed and isolated reticulorumen technique. Rumen papillae biopsies were used for adherent bacterial DNA and host RNA extraction. The epithelial bacteria were determined using Illumina MiSeq (Microsynth AG, Balgach, Switzerland) sequencing of the 16S rRNA gene. No significant effects of the high-grain feeding model were seen for bacterial diversity. However, bacterial diversity increased with time spent in the recovery forage feeding period regardless of feeding model. The relative abundance of Acidobacteria phyla and Acetivibrio spp. increased when animals were fed a transient high-grain feeding model. A trend toward increased CLDN4 expression was observed in the continuous model. Furthermore, there were interactions between feeding model and sampling day for gene targets CD14, DRA, NHE2, NHE3, and MCT2. When comparing length of recovery, in the continuous model increased relative absorption of SCFA was sustained at 63 d but dropped to baseline measurements at 105 d. A similar pattern was found with the transient model but it did not reach significance. The only gene target that was found to significantly correlate to relative absorption of SCFA was DRA (correlation coefficient ≤ -0.41). Whereas, genera Alkalibaculum, Anaerorhabdus, Coprococcus, and Dethiobacter all showed positive correlations to gene targets for pH regulation (NHE2 and NHE3) and SCFA uptake (MCT1) but negative correlations to SCFA absorption. We conclude that while the rumen absorption and epithelial bacteria were able to recover to baseline levels after 8 wk of forage feeding, the time needed for re-establishment of homeostasis in host gene expression is longer, especially when high-grain feeding is interrupted.

摘要

奶牛饲喂高谷物日粮通常会导致 pH 值降低和瘤胃微生物失调,其特征是短链脂肪酸 (SCFA) 在网胃壁的吸收动力学、上皮功能和上皮细菌群落结构发生变化。因此,本研究评估了高谷物持续喂养对恢复高谷物喂养的奶牛的网胃 SCFA 吸收动力学、瘤胃上皮基因表达以及相关上皮细菌变化的影响,这些奶牛来自长期持续高谷物喂养模型或长期短暂高谷物喂养模型。在交叉研究设计中,8 头非泌乳荷斯坦奶牛连续 4 周饲喂 60%浓缩饲料(连续)或在第二周的高谷物喂养中休息 1 周(短暂)。高谷物喂养后,所有动物均额外饲喂 100%饲草(恢复)饲料 8 周。在试验开始时(基线)、4 周高谷物喂养后(49 天)、2 周恢复饲草喂养后(63 天)和 8 周恢复饲草后(105 天)进行瘤胃乳头活检和 SCFA 吸收测量。使用冲洗和分离的网胃技术在体内确定 SCFA 的吸收。使用 Illumina MiSeq(Microsynth AG,Balgach,瑞士)对 16S rRNA 基因进行测序,从瘤胃乳头活检中提取粘附细菌 DNA 和宿主 RNA。上皮细菌用 16S rRNA 基因进行测定。高谷物喂养模型对细菌多样性没有显著影响。然而,无论喂养模型如何,在恢复饲草喂养期间,细菌多样性随着时间的推移而增加。当动物饲喂短暂的高谷物喂养模型时,厚壁菌门和乙酸菌属的相对丰度增加。在连续模型中观察到 CLDN4 表达呈上升趋势。此外,在基因靶标 CD14、DRA、NHE2、NHE3 和 MCT2 方面,还存在喂养模型和采样日之间的相互作用。当比较恢复时间时,在连续模型中,SCFA 的相对吸收增加在 63 天持续,但在 105 天降至基线测量值。在短暂模型中也发现了类似的模式,但没有达到显著水平。唯一发现与 SCFA 相对吸收显著相关的基因靶标是 DRA(相关系数≤-0.41)。然而,Alkalibaculum、Anaerorhabdus、Coprococcus 和 Dethiobacter 等属均与 pH 调节(NHE2 和 NHE3)和 SCFA 摄取(MCT1)的基因靶标呈正相关,但与 SCFA 吸收呈负相关。我们得出结论,尽管在 8 周的饲草喂养后,瘤胃吸收和上皮细菌能够恢复到基线水平,但宿主基因表达恢复内稳态所需的时间更长,尤其是当高谷物喂养中断时。

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