动物营养学报 >

2019 , Vol. 31 >Issue 1: 314 - 323

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.01.038

饲料科学与技术 Feed science and technology

采用Illumina MiSeq测序技术分析葡萄籽原花青素对奶牛体外瘤胃发酵产甲烷菌区系的影响

展开
  • 1. 奶牛营养学北京市重点实验室, 北京农学院动物科技学院, 北京 102206;
    2. 中国农业科学院北京畜牧兽医研究所, 北京 100193
童津津(1985-),女,黑龙江哈尔滨人,博士后,研究方向为奶牛营养与免疫。E-mail:tongjinjin0451@163.com

收稿日期: 2018-06-13

  网络出版日期: 2019-01-16

基金资助

国家自然科学基金项目（31772629，31702302）；2015年大北农青年教师科研基金；2016年北京农学院青年教师科研基金；2017年北京市教委科技计划项目（SQKM201710020011）；"十三五"国家重大科技专项（2016YFDO700201）；北京市农业局北京市现代农业产业技术体系奶牛创新团队

收起

Effects of Grape Seed Procyanidine on Methanogens Flora of in Vitro Rumen Fermentation of Dairy Cows Using Illumina MiSeq Sequencing Technology

Expand
  • 1. Beijing Key Laboratory for Dairy Cow Nutrition, Beijing University of Agriculture, Beijing 102206, China;
    2. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received date: 2018-06-13

  Online published: 2019-01-16

Fold

摘要

本研究旨在基于Illumina MiSeq测序技术分析葡萄籽原花青素对奶牛体外瘤胃发酵产甲烷菌区系的影响。试验分为6组，以500 g精粗比为40：60的全混合日粮为发酵底物，分别添加0（对照）、0.1、0.2、0.3、0.4、0.5 g/kg的葡萄籽原花青素。体外发酵24 h后提取各发酵液总DNA，采用Illumina MiSeq PE300高通量测序平台进行测序。结果表明：1）与对照组相比，添加0.1、0.2 g/kg葡萄籽原花青素降低了产甲烷菌的分类操作单元（OTUs）数量，添加0.3、0.4、0.5 g/kg葡萄籽原花青素增加了产甲烷菌的OTUs数量，但差异并不显著（P>0.05）；葡萄籽原花青素添加量对Chao1指数和Shannon指数均无显著影响（P>0.05），即对产甲烷菌的多样性没有显著影响。2）在门水平上，与对照组相比，添加0.3、0.4、0.5 g/kg葡萄籽原花青素显著降低了广古菌门的丰度（P<0.05）。在科水平上，与对照组相比，甲烷杆菌科的丰度随着葡萄籽原花青素添加量的增加有降低的趋势，且在添加量为0.4 g/kg时丰度显著降低（P<0.05）；Methanomassiliicoccaceae的丰度随着葡萄籽原花青素添加量的增加而增加，且当添加量为0.3、0.4 g/kg时显著增加（P<0.05）。在属水平上，与对照组相比，甲烷短杆菌属的丰度随着葡萄籽原花青素添加量的增加而降低，且当添加量为0.4 g/kg时显著降低（P<0.05），当添加量为0.5 g/kg时有回升的趋势，但是仍低于对照组；甲烷杆菌属的丰度随着葡萄籽原花青素添加量的增加而降低，且当添加量为0.4、0.5 g/kg时显著降低了该属的丰度（P<0.05）；Candidatus_Methanoplasma的丰度随着葡萄籽原花青素添加量的增加有上升趋势，且当添加量为0.3、0.4 g/kg时显著增加了该属的丰度（P<0.05）；甲烷球形菌属的丰度随着葡萄籽原花青素添加量的增加而降低，且当添加量为0.3、0.4、0.5 g/kg时显著降低了该属的丰度（P<0.05）。由此可知，添加葡萄籽原花青素在门、科、属水平上均对奶牛体外瘤胃产甲烷菌区系有一定的影响。

关键词： 葡萄籽原花青素; 奶牛; 产甲烷菌; Illumina MiSeq测序技术

本文引用格式

童津津, 张华, 孙铭维, 杨德莲, 张婕, 熊本海, 蒋林树 . 采用Illumina MiSeq测序技术分析葡萄籽原花青素对奶牛体外瘤胃发酵产甲烷菌区系的影响[J]. 动物营养学报, 2019 , 31(1) : 314 -323 . DOI: 10.3969/j.issn.1006-267x.2019.01.038

Abstract

The present study was designed to explore effects of grape seed procyanidine on Methanogens flora of in vitro rumen fermentation of dairy cows using Illumina MiSeq sequencing technology. The trial was divided into 6 groups. Total mixed ration with the concentrate to forage ratio at 40:60 was used as the fermentation substrate and added grape seed procyanidin at levels of 0 (control), 0.1, 0.2, 0.3, 0.4 and 0.5 g/kg, respectively. Total DNA was extracted from fermentation liquor after 24 h fermentation, and sequenced by Illumina MiSeq PE300 high-throughput sequencing platform. The results showed as follows:1) compared with the control group, the supplementation of grape seed procyanidins at 0.1, 0.2 g/kg significantly decreased the operational taxonomic units (OTUs), while the supplementation of grape seed procyanidins at 0.3, 0.4 and 0.5 g/kg increased the OTUs, but did not have significant difference (P>0.05); the supplementation of grape seed procyanidins did not significantly affect Chao1 index and Shannon index (P>0.05), namely did not significantly affect the diversity of Methanogens. 2) At phyla level, compared with the control group, the supplementation of grape seed procyanidine at 0.3, 0.4 and 0.5 g/kg significantly decreased the abundance of Euryarchaeota (P<0.05). At family level, compared with the control group, the abundance of Methanobacteriaceae had a decreased trend with the supplementation of grape seed procyanidine increasing, and when the supplementation was at 0.4 g/kg, the abundance of it significantly decreased (P<0.05); the abundance of Methanomassiliicoccaceae increased with the supplementation of grape seed procyanidine increasing, and when the supplementation was 0.3 and 0.4 g/kg, the abundance of it significantly increased (P<0.05). At genus level, compared with the control group, the abundance of Methanobrevibacter decreased with the supplementation of grape seed procyanidine increasing, and when the supplementation was 0.4 g/kg, the abundance of it significantly decreased (P<0.05), when the supplementation was 0.5 g/kg, the abundance of it had an increasing trend, but was lower than that in control group; the abundance of Methanobacterium decreased with the supplementation of grape seed procyanidine increasing, and when the supplementation was 0.4 and 0.5 g/kg, the abundance of it significantly decreased (P<0.05); the abundance of Candidatus_Methanoplasma had an increasing trend with the supplementation of grape seed procyanidine increasing, and when supplementation was 0.3 and 0.4 g/kg, the abundance of it significantly increased (P<0.05); the abundance of Methanosphaera decreased with the supplementation of grape seed procyanidine increasing, and when the supplementation was 0.3, 0.4 and 0.5 g/kg, the abundance of it significantly decreased (P<0.05). Therefore, the supplementation of grape seed procyanidine has significant effects on Methanogens at phyla, family and genus levels.

Key words： grape seed procyanidins; dairy cows; Methanogens; Illumina MiSeq sequencing technology

参考文献

[1] GILL M,SMITH P,WILKINSON J M.Mitigating climate change:the role of domestic livestock[J].Animal:An International Journal of Animal Bioscience,2010,4(3):323-333.  

[2] 周怿,刁其玉.反刍动物瘤胃甲烷气体生成的调控[J].草食家畜,2008(4):21-24.

[3] MOSS A R,JOUANY J P,NEWBOLD J.Methane production by ruminants:its contribution to global warming[J].Animal de Zootechnie,2000,49(3):231-253.  

[4] 游玉波.肉牛甲烷排放测定与估算模型的研究[D].博士学位论文.北京:中国农业科学院,2008.

[5] 娜仁花,董红敏.营养因素对反刍动物甲烷排放影响的研究现状[J].安徽农业科学,2009,37(6):2534-2536,2735.

[6] CAO Y,TAKAHASHI T,HORIGUCHI K I,et al.Methane emissions from sheep fed fermented or non-fermented total mixed ration containing whole-crop rice and rice bran[J].Animal Feed Science and Technology,2010,157(1/2):72-78.

[7] 严淑红,蒋林树.茶皂素对反刍动物瘤胃发酵的影响研究进展[J].中国农学通报,2015,31(5):20-24.

[8] 胡伟莲.皂甙对瘤胃发酵与甲烷产量及动物生产性能影响的研究[D].博士学位论文.杭州:浙江大学,2005.

[9] ODONGO N E,BAGG R,VESSIE G,et al.Long-term effects of feeding monensin on methane production in lactating dairy cows[J].Journal of Dairy Science,2007,90(4):1781-1788.  

[10] WANG J K,YE J A,LIU J X.Effects of tea saponins on rumen microbiota,rumen fermentation,methane production and growth performance-a review[J].Tropical Animal Health and Production,2012,44(4):697-706.  

[11] GEMEDA B S,HASSEN A.Effect of tannin and species variation on in vitro digestibility,gas,and methane production of tropical browse plants[J].Asian-Australasian Journal of Animal Sciences,2015,28(2):188-199.

[12] BAYATLI F,AKKU? D,KILIC E,et al.The protective effects of grape seed extract on MDA,AOPP,apoptosis and eNOS expression in testicular torsion:an experimental study[J].World Journal of Urology,2013,31(3):615-622.  

[13] GESSNER D K,WINKLER A,KOCH C,et al.Analysis of hepatic transcript profile and plasma lipid profile in early lactating dairy cows fed grape seed and grape marc meal extract[J].BMC Genomics,2017,18(1):253.

[14] 高巍,卜登攀,李永刚,等.葡萄籽粕对反刍动物营养价值的综合评定研究[C]//中国畜牧兽医学会动物营养学分会第七届中国饲料营养学术研讨会论文集.郑州:中国畜牧兽医学会动物营养学分会,2014.

[15] 周敏,叶子弘,蒋林树.瘤胃氢化多不饱和脂肪酸的影响因素[J].中国农学通报,2010,26(8):38-44.

[16] 杨德莲,童津津,张婕,等.葡萄籽原花青素对奶牛瘤胃体外发酵参数及微生物区系的影响[J].动物营养学报,2018,30(2):717-725.

[17] BVRGMANN H,PESARO M,WIDMER F,et al.A strategy for optimizing quality and quantity of DNA extracted from soil[J].Journal of Microbiological Methods,2001,45(1):7-20.  

[18] CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QⅡME allows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.  

[19] DRAY S,DUFOUR A,LEEUW J D,et al.The ade4 package:implementing the duality diagram for ecologists[J].Journal of Statistical Software,2007,22(4):1-20.

[20] 邸凌峰,曹雪,秦炜赜,等.两种含单宁饲草对反刍动物应用的价值研究[J].饲料工业,2017,38(7):43-50.

[21] 李晓鹏.缩合单宁对体外发酵特性的影响[D].硕士学位论文.兰州:甘肃农业大学,2009.

[22] 王慧玲,王小平,尕藏桑智,等.单宁酸对绵羊瘤胃体外发酵和甲烷产量的影响[J].中国草食动物科学,2013,33(6):46-48.

[23] ANANTASOOK N.日粮添加含单宁和皂苷的雨树荚对奶牛瘤胃发酵和甲烷产量调控的影响[J].王聪译.中国畜牧兽医,2014,41(2):152.

[24] HOOK S E,STEELE M A,NORTHWOOD K S,et al.Impact of high-concentrate feeding and low ruminal pH on methanogens and protozoa in the rumen of dairy cows[J].Microbial Ecology,2011,62(1):94-105.  

[25] 李志鹏,刘晗璐,司华哲,等.不同粗饲料条件下梅花鹿瘤胃甲烷菌结构的比较分析[J].动物营养学报,2016,28(9):2911-2919.

[26] CERSOSIMO L M,LACHANCE H,ST-PIERRE B,et al.Examination of the rumen bacteria and methanogenic archaea of wild impalas (Aepyceros melampus melampus) from Pongola,South Africa[J].Microbial Ecology,2015,69(3):577-585.  

[27] TAJIMA K,NAGAMINE T,MATSUI H,et al.Phylogenetic analysis of archaeal 16S rRNA libraries from the rumen suggests the existence of a novel group of archaea not associated with known methanogens[J].FEMS Microbiology Letters,2001,200(1):67-72.  

[28] BORREL G,HARRIS H M B,TOTTEY W,et al.Genome sequence of "Candidatus Methanomethylophilus alvus" Mx1201,a methanogenic archaeon from the human gut belonging to a seventh order of methanogens[J].Journal of Bacteriology,2012,194(24):6944-6945.  

[29] POULSEN M,SCHWAB C,JENSEN B B,et al.Methylotrophic methanogenic thermoplasmata implicated in reduced methane emissions from bovine rumen[J].Nature Communications,2013,4:1428.

[30] 郭嫣秋,胡伟莲,刘建新.瘤胃甲烷菌及甲烷生成的调控[C]//全国反刍动物营养生理生化第三次学术会议论文摘要汇编.长春:中国畜牧兽医学会,2004:244-247.

[31] MUSTAPHA N A,SAKAI K,SHIRAI Y,et al.Impact of different antibiotics on methane production using waste-activated sludge:mechanisms and microbial community dynamics[J].Applied Microbiology and Biotechnology,2016,100(21):9355-9364.  

[32] GOOPY J P,DONALDSON A,HEGARTY R,et al.Low-methane yield sheep have smaller rumens and shorter rumen retention time[J].British Journal of Nutrition,2014,111(4):578-585.  

[33] KING E E,SMITH R P,ST-PIERRE B,et al.Differences in the rumen methanogen populations of lactating jersey and Holstein dairy cows under the same diet regimen[J].Applied and Environmental Microbiology,2011,77(16):5682-5687.  

[34] CHANDRASEKHARAIAH M,THULASI A,JOSE L.Effect of supplementation of different essential oils on in-vitro methanogenesis,fermentation and digestibility of finger millet straw based diet in rumen liquor of crossbred cattle[J].Research Journal of Pharmaceutical,Biological and Chemical Sciences,2015,6(2):480-486.

[35] DANIELSSON R,DICKSVED J,SUN L,et al.Methane production in dairy cows correlates with rumen methanogenic and bacterial community structure[J].Frontiers in Microbiology,2017,8:226.

[36] 陈亚迎,刘壮,吕朋安,等.溶菌酶对瘤胃体外发酵、甲烷生成及微生物菌群结构的影响[J].微生物学报,2017,57(5):758-768.

[37] ST-PIERRE B,WRIGHT A D.Diversity of gut methanogens in herbivorous animals[J].Animal,2013,7(Suppl.1):49-56.

[38] 林波,梁辛,韦升菊,等.烟酸对水牛瘤胃发酵指标及微生物区系的影响[J].动物营养学报,2015,27(8):2396-2404.

[39] FRANZOLIN R,ST-PIERRE B,NORTHWOOD K,et al.Analysis of rumen methanogen diversity in water buffaloes (Bubalus bubalis) under three different diets[J].Microbial Ecology,2012,64(1):131-139.  

[40] 吴建敏,承尧兴,徐俊,等.葡萄籽残渣饲喂奶牛的效果研究[J].中国畜牧杂志,2007,43(7):62-63.

[41] 孙占鹏,王曦,李会菊,等.日粮中添加不同剂量葡萄渣对成年羊增重效果的影响[J].中国草食动物科学,2010,30(1):46-48.

[42] 赵栋,郑琛,李发弟,等.葡萄渣单宁对绵羊养分消化代谢及瘤胃发酵的影响[J].草业学报,2014,23(4):285-292.

[43] 李会菊,孙占鹏.日粮中添加葡萄渣对小尾寒羊成年母羊体重的影响[J].饲料工业,2010,31(3):33-36.
Options
文章导航

/

〈 〉

    联系我们

  • 地址：北京市海淀区圆明园西路2号
    　　　中国农业大学动科动医大楼153室
  • 邮编：100193
  • 电话：010－62817823
  • 网址：http://www.chinajan.com
  • E-mail：cjan@chinajan.com
京公网安备 11010802041926号    京ICP备17050989号-2