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白藜芦醇对不同类型底物甲烷产生、养分降解及微生物区系的影响
引用本文:吴万成,马涛,刘娜,杨磊,陈国顺,刁其玉.白藜芦醇对不同类型底物甲烷产生、养分降解及微生物区系的影响[J].畜牧兽医学报,2020,51(6):1281-1294.
作者姓名:吴万成  马涛  刘娜  杨磊  陈国顺  刁其玉
作者单位:1. 中国农业科学院饲料研究所, 农业农村部饲料生物技术重点实验室, 北京 100081;2. 甘肃农业大学动物科学技术学院, 兰州 730070
基金项目:国家自然科学基金(41705129);国家重点研发计划“反刍动物甲烷排放测算模型及基于宏基因组学的减排技术”(2016YFE0109000)
摘    要:本试验研究了白藜芦醇(RES)对两种类型底物瘤胃发酵的甲烷(CH 4)排放、养分降解以及微生物区系影响,旨在探索RES调控瘤胃发酵、降低CH 4排放的机理。本试验为双因素设计,选取3只健康、体重相近((50±2.3)kg)且安装有永久性瘤胃瘘管的杜寒杂交肉羊做为瘤胃液供体。试验一,在精粗比为68∶32(高精料,HC)和28∶72(高粗料,HF)的两种底物中分别添加0%(对照组)、7.7%、14.3%和25.0%的RES进行产气试验,并选取对照组和14.3%组进行16S扩增子高通量测序;试验二,在上述两种底物中分别添加0%、14.3%的RES进行降解率试验。以上试验处理均设置5个重复。试验结果如下:1)两种底物产气量(GP)和CH 4产量随RES水平的增加均线性降低(P<0.05);在相同RES处理下,相较HC,HF中GP和CH 4产量均显著降低(P<0.05);且两因素对两指标变化存在交互效应(P<0.05)。2)添加RES后两种底物干物质(DM)、有机物(OM)、粗蛋白质(CP)、中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)降解率均显著降低(P<0.05);在相同RES处理下,相较HC,HF中各养分降解率均显著降低(P<0.05),且两因素对发酵24 h中性洗涤纤维降解率(NDFD)和酸性洗涤纤维降解率(ADFD)存在交互作用(P<0.05)。3)从门水平来看,两种底物中添加RES均引起变形菌门(Proteobacteria)丰度显著升高(P<0.05);相较HC,HF的变形菌门丰度显著升高(P<0.05)。另外,在发酵第24小时,HF中添加RES添加导致互养菌门显著降低(P<0.05);发酵第12小时,同一RES处理水平下,相较HC,HF中放线菌门丰度显著降低。从属水平来看,添加RES和改变底物类型均导致Prevotella_1(普雷沃氏菌_1)丰度大幅下降,且在HC(发酵12 h)、HF(发酵24 h)具有统计学意义(P<0.05);添加RES变形菌门的优势菌属Ruminobacter和Succinivibrio均显著增加(P<0.05)。另外,发酵第12小时,在HF添加RES,Rikenellaceae_RC9_gut_group、Prevotellaceae_NK3B31_group和Christensenellaceae_R-7_group丰度均显著上升(P<0.05);相较HC,HF中Prevotellaceae_UCG-001、Prevotellaceae_NK3B31_group和Christensenellaceae_R-7_group丰度显著降低(P<0.05)。在发酵第24小时,在HF中添加RES导致Veillonellaceae_UCG-001和Lachnospiraceae_XPB1014_group丰度显著降低(P<0.05),Prevotellaceae_UCG-001、Phocaeicola、Prevotellaceae_NK3B31_group、Succiniclasticum、Christensenellaceae_R-7_group、Ruminococcaceae_NK4A214_group和Ruminobacter丰度显著升高(P<0.05);且两因素对普雷沃氏菌_1、Succiniclasticum、Veillonellacea e_UCG-001和Lachnospiraceae_XPB1014_group的丰度变化存在互作效应(P<0.05)。此结果说明,添加高水平的RES可以有效降低瘤胃环境CH 4的排放量,但是抑制了各养分在瘤胃中的降解,这可能与瘤胃菌群中普雷沃氏菌_1的大幅度下降有关。

关 键 词:白藜芦醇  甲烷  降解率  群落丰度
收稿时间:2019-10-21

Effect of Resveratrol on Methane Production,Nutrient Degradation and Microbial Community under Different Substrates
WU Wancheng,MA Tao,LIU Na,YANG Lei,CHEN Guoshun,DIAO Qiyu.Effect of Resveratrol on Methane Production,Nutrient Degradation and Microbial Community under Different Substrates[J].Acta Veterinaria et Zootechnica Sinica,2020,51(6):1281-1294.
Authors:WU Wancheng  MA Tao  LIU Na  YANG Lei  CHEN Guoshun  DIAO Qiyu
Institution:1. Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Feed Research Institute of Chinese Academy of Agricultural Sciences, Beijing 100081, China;2. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
Abstract:This experiment investigated the effects of resveratrol (RES) on methane (CH4) emissions, nutrient degradation and microbial community in two types of substrate, aiming to explore the mechanisms of RES regulating rumen fermentation and reducing CH4 emissions. This experiment was a two-factor design. Three rams with similar body weight((50±2.3) kg) and health status installed with permanent rumen fistula were selected as rumen fluid donors. In experiment one, 0%, 7.7%, 14.3% and 25.0% RES were added to the substrates with a concentrate:forage ratio of 68:32 (high concentrate, HC) and 28:72 (high forage, HF), respectively, for testing gas production. High-throughput sequencing of 16S amplicons was performed in the control group and 14.3% group. In experiment two, 0% and 14.3% RES were added to the above two substrates for degradation rate test. Five replicates were set for the above experimental treatments. The results showed that:1) The gas production (GP) and CH4 production of the two substrates decreased linearly with the increase in RES level (P<0.05); At the same RES level, compared with HC, GP and CH4 production in HF were significantly reduced (P<0.05), and the two factors had an interactive effect on the changes of the two indicators (P<0.05). 2) The degradation rates of dry matter(DM), organic matter(OM), crude protein(CP), neutral detergent fiber(NDF), and acid detergent fiber(ADF) of the two substrates were significantly decreased after supplemented with RES (P<0.05). Compared with HC, the degradation rate of each nutrient in HF was significantly reduced at the same RES level, and there was an interaction between the two factors on degradation rate of neutral detergent fiber (NDFD) and degradation rate of acid detergent fiber (ADFD) at 24 h of fermentation (P<0.05). 3) At phylum level, the addition of RES in both substrates resulted in a significant increase in the Proteobacteria (P<0.05); compared with HC, the abundance of the Proteobacteria in HF increased significantly (P<0.05). In addition, at 24 h of fermentation, the addition of RES in HF caused a significant decrease in the Synergistetes (P<0.05); at 12 h of fermentation, Compared with HC, the abundance of Actinomycetes in HF significantly decreased at the same RES level. At genus level, the addition of RES and the change of substrate types all resulted in a significant decrease in the abundance of Prevotella_1, and it was statistically significant at HC (12 h fermentation) and HF (24 h fermentation) (P<0.05); Ruminobacter and Succinivibrio, the dominant strains of Proteobacteria, increased significantly with RES supplemetation (P<0.05). In addition, at 12 h of fermentation, RES was added to HF, the abundance of Rikenellaceae_RC9_gut_group, Prevotellaceae_NK3B31_group and Christensenellaceae_R-7_group increased significantly (P<0.05). Compared with HC, Prevotellaceae_UCG-001, Prevotellaceae_NK3B31_group and Christensen ellaceae_R-7-_group decreased significantly in HF (P<0.05). At 24 h of fermentation, the addition of RES in HF resulted in a significant decrease in the abundance of Veillonellaceae_UCG-001 and Lachnospiraceae_XPB1014_group (P<0.05), Prevotellaceae_UCG-001, Phocaeicola, Prevotellaceae_NK3B31_group, Succiniclasticum, Christensenellaceae_R-7_group, Ruminococcaceae_NK4A214_group and Ruminobacter were significantly increased (P<0.05); and the two factors had an interaction effect on the abundance changes of Prevotella_1, Succiniclasticum, Veillonellaceae_UCG-001 and Lachnospiraceae_XPB1014_group (P<0.05). The results showed that,high level of RES suppplementation effectively reduced the CH4 emission, but inhibited the degradation of nutrient in the rumen, which might be related to the significant decline of Prevotella_1 in the rumen flora.
Keywords:resveratrol  methane  degradation rate  community abundance  
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