研究论文 RESEARCH PAPER

猪粪便微生物移植小鼠肠道菌群结构演替分析

  • 宋园园 ,
  • 施伟领 ,
  • 章啸君 ,
  • 杨华 ,
  • 吴酬飞 ,
  • 肖英平
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  • 1. 湖州师范学院生命科学学院, 湖州 313000;
    2. 浙江农业科学院省部共建农产品质量安全危害因子与风险防控国家重点实验室, 农产品质量安全与营养研究所, 杭州 310021;
    3. 浙江大飞龙动物保健品有限公司, 金华 321017;
    4. 金华市农业科学研究院畜牧兽医研究所, 金华 321017
宋园园(1997-),女,河南焦作人,硕士研究生,研究方向为畜禽肠道微生物。E-mail:songyuanyuan0215@126.com

收稿日期: 2022-02-26

  网络出版日期: 2022-10-17

基金资助

国家自然科学基金(31972999)

Succession Analysis of Intestinal Microbiota Structure of Mice with Pig Fecal Microbiota Transplantation

  • SONG Yuanyuan ,
  • SHI Weiling ,
  • ZHANG Xiaojun ,
  • YANG Hua ,
  • WU Choufei ,
  • XIAO Yingping
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  • 1. College of Life Science, Huzhou University, Huzhou 313000, China;
    2. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Agro-Product Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
    3. Zhejiang Dovro Animal Health Business Company, Jinhua 321017, China;
    4. Institute of Animal Husbandry and Veterinary Medicine, Jinhua Academy of Agricultural Sciences, Jinhua 321017, China

Received date: 2022-02-26

  Online published: 2022-10-17

摘要

本试验旨在探究猪粪便微生物移植对受体小鼠肠道菌群结构的动态演替变化。选择金华猪和长白猪作为粪便移植供体,通过灌胃将猪粪便微生物移植于经广谱抗生素处理过的小鼠体内,连续灌胃7 d,在灌胃第1、2、4和7天以及在停止灌胃后第7、14、21和28天收集小鼠粪便,分别采用16S rRNA基因测序、实时荧光定量PCR(RT-PCR)和气相色谱法测定小鼠肠道菌群结构和数量的动态变化以及短链脂肪酸(SCFAs)含量变化。结果表明:在猪到小鼠的粪便移植过程中,受体小鼠粪便样品中微生物数量和alpha多样性随移植时间延长呈上升趋势。在门水平上,随着移植时间的延长,受体小鼠肠道中厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)相对丰度逐渐降低,而拟杆菌门(Bacteroidetes)和疣微菌门(Verrucomicrobia)相对丰度逐渐升高;在属水平上,拟杆菌属(Bacteroides)、阿克曼氏菌属(Akkermansia)和副拟杆菌属(Parabacteroides)相对丰度逐渐升高,而乳酸杆菌属(Lactobacillus)、肠球菌属(Enterococcus)、大肠杆菌属(Escherichia)的相对丰度逐渐降低,直至停止移植后保持稳定。移植过程中猪粪便微生物受体小鼠粪便样品中SCFAs的含量随移植时间延长而逐渐升高,停止移植后SCFAs含量变化较小,并在停止移植后第28天基本保持稳定。结果提示,猪粪便微生物跨物种移植于抗生素处理小鼠,可以使受体小鼠肠道菌群出现动态演替性变化,且不同品种猪粪便移植小鼠肠道菌群结构受猪供体微生物影响而有所不同,菌群代谢产生SCFAs的含量也表现出与菌群数量相一致的变化规律。

本文引用格式

宋园园 , 施伟领 , 章啸君 , 杨华 , 吴酬飞 , 肖英平 . 猪粪便微生物移植小鼠肠道菌群结构演替分析[J]. 动物营养学报, 2022 , 34(10) : 6758 -6767 . DOI: 10.3969/j.issn.1006-267x.2022.10.068

Abstract

This experiment was conducted to explore the dynamic succession of the intestinal microbiota structure of mice with pig fecal microbiota transplantation. Jinhua pigs and Landrace pigs were used as fecal transplant donors, and fecal microorganisms from pigs were administered into germ-free mice for 7 days after the treatment of broad spectrum antibiotics. The feces of mice were collected at days 1, 2, 4 and 7 during the administration and days 7, 14, 21 and 28 after the administration. The dynamic changes in the structure and quantity of mice intestinal microbiota and the changes in short-chain fatty acids (SCFAs) contents in the feces were determined by 16S rRNA gene sequencing, RT-PCR, and gas chromatography, respectively. The results showed that the number and alpha diversity of microorganisms in the fecal samples of recipient mice increased with transplantation time during pig fecal microbiota transplantation. At the phylum level, the relative abundance of Firmicutes and Proteobacteria in the intestine of recipient mice gradually decreased over the time of transplantation, while the relative abundance of Bacteroidetes and Verrucomicrobia increased; at the genus level, the relative abundance of Bacteroides, Akkermansia and Parabacteroides increased, while the relative abundance of Lactobacillus, Enterococcus and Escherichia decreased until reaching a stable platform after the transplantation. The content of SCFAs in the fecal samples of mice was gradually increased over the time of transplantation and the changes in the content of SCFAs became less after the transplantation and remained stable at day 21 after the transplantation. The results suggest that cross-species transplantation of pig fecal microbiota into antibiotic-treated mice can lead to dynamic succession in the intestinal intestinal microbiota of the recipient mice, the intestinal microbiota structure of mice with fecal microbiota transplantation from different pig breeds are different due to the influence of pig donor microorganisms and the contents of SCFAs produced by the metabolism of the intestinal microbiota also show a similar pattern of change in the number of intestinal microorganisms.

参考文献

[1] BÄCKHED F,MANCHESTER J K,SEMENKOVICH C F,et al.Mechanisms underlying the resistance to diet-induced obesity in germ-free mice[J].Proceedings of the National Academy of Sciences of the United States of America,2007,104(3):979-984.  
[2] 岳晓敬.粪菌移植对仔猪生产性能,肠道菌群和肠粘膜发育的影响[D].硕士学位论文.杭州:浙江大学,2017. YUE X J.Effects of fecal microbiota transplantation on piglet performance,intestinal flora and intestinal mucosal development[D].Master's Thesis.Hangzhou:Zhejiang University,2017.(in Chinese)
[3] CHUNG H,PAMP S J,HILL J A,et al.Gut immune maturation depends on colonization with a host-specific microbiota[J].Cell,2012,149(7):1578-1593.  
[4] VALERIANO V D V,BALOLONG M P,KANG D K.Probiotic roles of Lactobacillus sp.in swine:insights from gut microbiota[J].Journal of Applied Microbiology,2017,122(3):554-567.  
[5] BÄCKHED F,DING H,WANG T,et al.The gut microbiota as an environmental factor that regulates fat storage[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(44):15718-15723.  
[6] CAMMAROTA G,IANIRO G,TILG H,et al.European consensus conference on faecal microbiota transplantation in clinical practice[J].Gut,2017,66(4):569-580.  
[7] DREW L.Microbiota:reseeding the gut[J].Nature,2016,540(7634):S109-S112.
[8] 陈佳琦,张晓迪,顾招兵,等.粪便微生物移植对仔猪肠道屏障功能的影响及作用机制进展[J].畜牧兽医学报,2021,52(5):1195-1207. CHEN J Q,ZHANG X D,GU Z B,et al.Effects and mechanism of faecal microbiota transplantation on intestinal barrier function in piglets[J].Acta Veterinaria et Zootechnica Sinica,2021,52(5):1195-1207.(in Chinese)
[9] YANG B,TIAN H L,YE C,et al.The efficacy and safety of fecal microbiota transplantation combined with biofeedback for mixed constipation:a retrospective cohort study[J].Frontiers in Medicine,2021,8:746990.
[10] SERRANO-VILLAR S,TALAVERA-RODRÍGUEZ A,GOSALBES M J,et al.Fecal microbiota transplantation in HIV:a pilot placebo-controlled study[J].Nature Communications,2021,12(1):1139.
[11] 夏戴阳,杨琳,朱勇文,等.鸭粪便微生物移植技术[J].动物营养学报,2021,33(1):95-100. XIA D Y,YANG L,ZHU Y W,et al.Fecal microbial transplantation technology for ducks[J].Chinese Journal of Animal Nutrition,2021,33(1):95-100.(in Chinese)
[12] COLLINS S M,KASSAM Z,BERCIK P.The adoptive transfer of behavioral phenotype via the intestinal microbiota:experimental evidence and clinical implications[J].Current Opinion in Microbiology,2013,16(3):240-245.  
[13] WU C,LYU W,HONG Q,et al.Gut microbiota influence lipid metabolism of skeletal muscle in pigs[J].Fronties in Nutrition,2021,8:675445.
[14] YAN H L,DIAO H,XIAO Y,et al.Gut microbiota can transfer fiber characteristics and lipid metabolic profiles of skeletal muscle from pigs to germ-free mice[J].Scientific Reports,2016,6:31786.
[15] DIAO H,YAN H L,XIAO Y,et al.Intestinal microbiota could transfer host gut characteristics from pigs to mice[J].BMC Microbiology,2016,16(1):238.
[16] LI N,ZUO B,HUANG S M,et al.Spatial heterogeneity of bacterial colonization across different gut segments following inter-species microbiota transplantation[J].Microbiome,2020,8(1):161.
[17] AGGARWALA V,MOGNO I,LI Z H,et al.Precise quantification of bacterial strains after fecal microbiota transplantation delineates long-term engraftment and explains outcomes[J].Nature Microbiology,2021,6(10):1309-1318.  
[18] YANG H,XIANG Y,ROBINSON K,et al.Gut microbiota is a major contributor to adiposity in pigs[J].Frontiers in Microbiology,2018,9:3045.
[19] XU J,VERBRUGGHE A,LOURENCO M,et al.The response of canine faecal microbiota to increased dietary protein is influenced by body condition[J].BMC Veterinary Research,2017,13(1):374.
[20] HÅKANSSON Å,TORMO-BADIA N,BARIDI A,et al.Immunological alteration and changes of gut microbiota after dextran sulfate sodium (DSS) administration in mice[J].Clinical and Experimental Medicine,2015,15(1):107-120.  
[21] 郭慧玲,邵玉宇,孟和毕力格,等.肠道菌群与疾病关系的研究进展[J].微生物学通报,2015,42(2):400-410. GUO H L,SHAO Y Y,MENG H B L G,et al.Research on the relation between gastrointestinal microbiota and disease[J].Microbiology,2015,42(2):400-410.(in Chinese)
[22] 张卓.粪菌移植和结肠菌移植对新生仔猪生长、代谢及肠道健康的差异影响[D].硕士学位论文.重庆:西南大学,2021. ZHANG Z.Differential effects of fecal microbiota transplantation and colonic microbiota transplantation on growth,metabolism and intestinal health of neonatal piglets[D].Master's Thesis.Chongqing:Southwest University,2021.(in Chinese)
[23] GENG S J,CHENG S S,LI Y,et al.Faecal microbiota transplantation reduces susceptibility to epithelial injury and modulates tryptophan metabolism of the microbial community in a piglet model[J].Journal of Crohn's&Colitis,2018,12(11):1359-1374.
[24] 陈雪,任二都,苏勇.早期灌喂母源粪菌对新生仔猪肠道菌群发育的影响[J].微生物学报,2018,58(7):1224-1232. CHEN X,REN E D,SU Y.Effect of oral feeding maternal fecal microbiota on intestinal microbiota development of newborn piglets[J].Acta Microbiologica Sinica,2018,58(7):1224-1232.(in Chinese)
[25] GREHAN M J,BORODY T J,LEIS S M,et al.Durable alteration of the colonic microbiota by the administration of donor fecal flora[J].Journal of Clinical Gastroenterology,2010,44(8):551-561.  
[26] LI Y Z,CAO W M,GAO N L,et al.Consistent alterations of human fecal microbes after transplantation into germ-free mice[J/OL].Genomics Proteomics&Bioinformatics,2021.(2021-06-09)[2021-12-20].https://www.sciencedirect.com/science/article/pii/S167202292100098X.DOI:10.1016/j.gpb.2020.06.024.
[27] 杨华.金华猪肠道微生物结构解析及其与脂肪沉积的关联分析[D].博士学位论文.北京:中国农业大学,2018. YANG H.Gut microbial community and its correlation with fat deposition in Jinhua pigs[D]. Ph.D.Thesis.Beijing:China Agricultural University,2018.(in Chinese)
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