猪营养与饲料 SWINE NUTRITION AND FEED

哺乳仔猪断奶前后肠道微生物培养组学研究

  • 董博 ,
  • 王志林 ,
  • 魏文康 ,
  • 井晓欢 ,
  • 刘石 ,
  • 鄢秋龙 ,
  • 蒋宗勇 ,
  • 刘文华 ,
  • 胡译尹 ,
  • 陈庄
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  • 1. 广东省农业科学院农业生物基因研究中心, 广东省农作物种质资源保存与利用重点实验室, 广州 510640;
    2. 广东省畜禽疫病防治研究重点实验室, 农业农村部兽用药物与诊断技术广东科学观测实验站, 广州 510640;
    3. 岭南现代农业科学与技术广东省实验室茂名分中心, 茂名 525000;
    4. 深圳国家基因库, 深圳 510083;
    5. 大连医科大学微生物教研室, 大连 116044;
    6. 广东省农业科学院, 广州 510640
董博(1982-),男,湖北黄冈人,博士研究生,从事畜禽肠道微生物和营养研究。E-mail:dongbo@gdaas.cn

收稿日期: 2020-06-13

  网络出版日期: 2021-01-18

基金资助

广东省农业科学院院长基金项目(201814B,201723);广东省畜禽疫病防治研究重点实验室开放基金项目(YDWS1908);广东省科技计划(2014B020201002)

Study on Intestinal Microbial Culturomics of Piglets Before and After Weaning

  • DONG Bo ,
  • WANG Zhilin ,
  • WEI Wenkang ,
  • JING Xiaohuan ,
  • LIU Shi ,
  • YAN Qiulong ,
  • JIANG Zongyong ,
  • LIU Wenhua ,
  • HU Yiyin ,
  • CHEN Zhuang
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  • 1. Guangdong Key Laboratory of Crop Germplasm Resources Conservation and Utilization, Agro-Genes Research Center, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
    2. Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture and Rural Affairs, Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou 510640, China;
    3. Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology-Maoming Sub-Center, Maoming 525000, China;
    4. China National GeneBank, Shenzhen 510083, China;
    5. Department of Microbiology, Dalian Medical University, Dalian 116044, China;
    6. Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Received date: 2020-06-13

  Online published: 2021-01-18

摘要

近年来国内外研究人员通过下一代测序和分离培养技术,结合宏基因组学相关的生物信息学手段证实了猪肠道微生物的组成及其代谢物活性对宿主的健康具有重要作用。受限于传统培养方法,猪肠道中大多数细菌仍无法培养,因此,为了突破研究人员研究宿主-菌株相互关系及益生菌株开发应用的限制,本研究对仔猪断奶前后回肠和结肠内容物微生物进行高通量培养组学研究。结合需氧和厌氧条件、不同培养时间以及25种不同培养基,共筛选获得1 385株厌氧、好氧和兼性厌氧菌株,并全部进行了16S rRNA全长测序鉴定和菌株保存,共计获得5个门、29个属和86个种菌,其中包含梭杆菌、拟杆菌等在以前的研究中较难分离获得的菌株,以及一些如乳酸菌等具有潜在应用价值的益生菌,更为重要的是其中包含116株疑似新菌种。本研究获得的菌株信息可以进一步完善猪肠道微生物物种数据库,为后续宿主-菌株相互关系研究垫定了基础,对猪饲用益生菌相关产品开发研究具有重要意义。

本文引用格式

董博 , 王志林 , 魏文康 , 井晓欢 , 刘石 , 鄢秋龙 , 蒋宗勇 , 刘文华 , 胡译尹 , 陈庄 . 哺乳仔猪断奶前后肠道微生物培养组学研究[J]. 动物营养学报, 2021 , 33(1) : 175 -189 . DOI: 10.3969/j.issn.1006-267x.2021.01.019

Abstract

In recent years, researchers at home and abroad have proved that the composition of porcine intestinal microorganisms and their metabolite activity play an important role in host health through next-generation sequencing and isolation culture techniques combined with metagenomics related bioinformatics methods. Due to the limitations of traditional culture methods, most of the bacteria in the intestinal tract of pigs still cannot be cultured. Therefore, in order to break through the limitations of studying host-strain interaction and the development and application of probiotics, this study applied high-throughput culturomics to study the microorganisms in ileum and colon contents of piglets before and after weaning combined with aerobic and anaerobic conditions, different culture time and 25 different medium, then a total of 1 385 anaerobic, aerobic and facultative strains were screened, and for all these full-length 16S rRNA sequencing identification and strain preservation were carried out. A total of 5 phyla, 29 genera and 86 species were identified, including some strains which were difficult to cultivate using traditional culture techniques, such as Fusobacteria, Bacteroides, etc., and some probiotic bacteria with potential application value such as Lactobacillus was isolated; more importantly, they contained 116 suspected new strains. The strain information obtained in this study can further improve the database of intestinal microbial species of pigs, lay a foundation for subsequent studies on host-strain interaction, and is of great significance for the development and research of probiotics related products for pig feeding.

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