人工瘤胃模拟装置发酵内环境变化规律研究  

The Law of Environmental Variation in Fermentation of Artificial Rumen Simulator

作　　者：皇甫和平[1] 徐梓明 宋超 任晓丽 石冬梅[1,4] 董青 HUANGFU Heping;XU Ziming;SONG Chao;REN Xiaoli;SHI Dongmei;DONG Qing(College of Veterinary Medicine,Henan University of Animal Husbandry and Economy,Zhengzhou 450046;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306;Henan key Laboratory of Innovation and Utilization of Unconventional Feed Resources,Zhengzhiou 450046;Zhengzhou key Laboratory of Animal Nutrition Metabolic and Poisoning Diseases,Zhengzhou 450046)

机构地区：[1]河南牧业经济学院动物医药学院,河南郑州450046 [2]上海海洋大学水产与生命学院,上海201306 [3]河南省非常规饲料资源创新利用重点实验室,河南郑州450046 [4]郑州市动物营养代谢病与中毒病重点实验室,河南郑州450046

出　　处：《家畜生态学报》2023年第5期52-58,共7页Journal of Domestic Animal Ecology

基　　金：河南牧业经济学院校级重点学科建设项目(XJXK202202);河南省高等学校重点科研项目(20B230006);河南省科技攻关项目(212102110377、212102110372、232102111037)。

摘　　要：为了解人工瘤胃模拟装置发酵内环境变化规律,将新鲜采集的牛瘤胃液按比例按比例加入到人工瘤胃模拟装置中,分组发酵培养6 h、12 h、18 h和24 h后,分别测定pH及发酵产气量,并提取不同培养时长瘤胃液样本总DNA,使用Miseq2×300 bp平台对样本16S rRNA基因V3~V4、18S rRNA基因V4区扩增产物测序,进行微生物宏基因组学分析。结果表明:随着发酵时间延长,瘤胃pH不断下降,发酵总产气量整体呈上升趋势,且12 h内呈现显著性变化(P<0.05);针对瘤胃发酵液中原核型微生物和真核型微生物类群进行的宏基因组测序结果分析表明,发酵培养12 h的样本中微生物物种丰度及群落多样性指数处于较高水平,与0 h发酵样本差异显著(P<0.05),且随培养时间延长,群落多样性整体呈下降趋势,普雷沃菌属的物种丰度最高,均毛属为瘤胃纤毛虫优势种属。值得注意的是,瘤胃液中尚存在许多未被鉴别分类,且相对丰度较高的微生物,需要进一步研究。To study the variation pattern of the internal rumen environment using an artificial rumen simulator and METAGENOMIC technique,the fresh rumen fluid was proportionally added to the artificial rumen simulator;after 6 h,12 h,18 h and 24 h of fermentation culture in different groups,the pH value and fermentation gas production rate were measured.Total DNAs from rumen fluid samples cultured for different time lengths were extracted,and amplification products of 16S rRNA gene V3~V4 and 18S rRNA gene V4 regions of the samples were sequenced using Miseq2×300 bp platform.The results showed that the rumen pH value decreased with the prolongation of fermentation time,and the total gas production rate of fermentation showed an upward trend,and showed a significant change within 12 h(P<0.05).Analysis of metagenome sequencing results of prokaryotic microorganisms as well as eukaryotic microorganisms in rumen fermentation broth showed that the microbial species abundance and community diversity index in the samples cultured for 12 h were at a high level,which was significantly different from the samples fermented for 0 h(P<0.05),and the overall community diversity showed a downward trend with the extension of culture time.The species abundance of Prevotella was the highest,and the Isotricha was the dominant species of rumen flora.Notably,many unidentified microorganisms with high relative abundance were also found in rumen fluid that need further investigation.

关 键 词：人工瘤胃 宏基因组技术 瘤胃内环境 

分 类 号：S823.5[农业科学—畜牧学]