研究简报 Short Communications

运用454焦磷酸测序技术对断奶前后仔猪肠道菌群的分析

  • 王一冰 ,
  • 张小平 ,
  • 黄怡 ,
  • 李卫芬
展开
  • 1. 浙江大学动物科学学院, 饲料研究所, 杭州 310058;
    2. 广西大学动物科学技术学院, 南宁 530004

收稿日期: 2013-04-11

  网络出版日期: 2013-09-16

基金资助

国家863计划项目资助(2013AA102800);教育部博士点基金(20110101110101);浙江省科技厅重大专项(2006C12086)

Intestinal Microflora Analysis of Pre-and Post-Weaning Piglets Using by 454 Pyrosequencing Technology

  • WANG Yibing ,
  • ZHANG Xiaoping ,
  • HUANG Yi ,
  • LI Weifen
Expand
  • 1. Institute of Feed Science, College of Animal Science, Zhejiang University, Hangzhou 310058, China;
    2. College of Animal Science and Technology, Guangxi University, Nanning 530004, China

Received date: 2013-04-11

  Online published: 2013-09-16

摘要

本研究旨在运用454焦磷酸测序技术分析断奶前后仔猪肠道菌群的变化。随机选取胎次和出生时间相近的、体重差异不显著的健康"杜×长×大"外三元新生仔猪12头用于试验。整个试验期间由母猪按常规哺乳直到断奶(25日龄),母猪饲粮不含有抗生素,仔猪于12日龄开始饲喂教槽料。于25日龄(断奶前)和32日龄(断奶后)分别随机挑选3头仔猪进行心脏放血屠宰,无菌采集盲肠内容物,运用454焦磷酸测序技术分析断奶前后仔猪盲肠内容物中菌群结构的变化。结果表明:与断奶前仔猪相比,断奶后仔猪腹泻率显著增加(P<0.05);肠道中菌群多样性增加;肠道中拟杆菌门(Bacteroidetes)含量变化不显著(P>0.05),厚壁菌门(Firmicutes)含量显著增加了63.95%(P<0.05),梭杆菌门(Fusobacteria)和变形菌门(Proteobacteria)含量分别显著减少了100.00%和70.54%(P<0.05)。对厚壁菌门含量进行深入分析发现,在纲的水平,与断奶前仔猪相比,断奶后仔猪肠道中梭菌纲(Clostridia)和芽孢杆菌纲(Bacilli)含量有减少的趋势(P>0.05),而Negativicutes纲、Erysipelotrichia纲和未知厚壁菌纲含量有增加的趋势(P>0.05)。从门到属的水平,断奶前后仔猪肠道菌群相对丰度不同,有些菌是断奶前仔猪肠道特有的,如嗜胆菌属(Bilophila)、具核梭杆菌属(Fusobacterium nucleatum)、黄杆菌属(Flavobacteriaceae)等,有些菌是断奶后仔猪肠道特有的,如费克蓝姆菌属(Facklamia)、八叠球菌属(Sarcina)等。由此可见,断奶后仔猪肠道菌群多样性和菌群结构发生了改变。

本文引用格式

王一冰 , 张小平 , 黄怡 , 李卫芬 . 运用454焦磷酸测序技术对断奶前后仔猪肠道菌群的分析[J]. 动物营养学报, 2013 , 25(10) : 2440 -2446 . DOI: 10.3969/j.issn.1006-267x.2013.10.028

Abstract

This present study was designed to analyze the intestinal microflora change of pre-and post-weaning piglets using by 454 pyrosequencing technology. Twelve newborn piglets (Duroc×Landrace×Large White) with the similar parity, birth time, and body weight were used in this experiment. All the experimental piglets were conventionally suckled by sows until weaning (25 days of age). The diet of sows did not contain antibiotics, and all piglets had free access to pre-starter diets from 12 days of age. Three piglets were randomly selected and killed at 25 (pre-weaning) and 32 (post-weaning) days of age, and cecal contents were collected. Microflora structure of cecal contents of pre-and post-weaning piglets were evaluated using by 454 pyrosequencing technology. The results showed that compared with the pre-weaning piglets, the diarrhea rate of post-weaning piglets was significantly increased (P<0.05); the diversity of intestinal microflora was increased; no difference was observed in the content of Bacteroides of post-weaning piglets (P>0.05); the content of Firmicutes was significantly increased by 63.95% (P<0.05), while the contents of Fusobacteria and Proteobacteria were significantly decreased by 100.00% and 70.54% (P<0.05). For Firmicutes, a further analysis was made. At class level, the contents of Bacilli and Negativicutes of post-weaning piglets were decreased (P>0.05), while the contents of Negativicutes, Erysipelotrichia and unclassified Firmicutes were increased (P>0.05). The relative abundance of each species from phylum to genus was different between pre-and post-weaning piglets. Some genera (such as Bilophila, Fusobacteriu and Flavobacteriaceae, etc.) only appeared in the pre-weaning piglets, while some other genera(such as Facklami and Sarcina, etc.)existed only in the post-weaning piglets. In conclusion, the diversity and the composition of intestinal microflora are altered in post-weaning piglets.

参考文献

[1] BROOM L J,MILLER H M,KERR K G,et al.Effects of zinc oxide and Enterococcus faecium SF68 dietary supplementation on the performance,intestinal microbiota and immune status of weaned piglets[J].Research in Veterinary Science,2006,80(1):45-54.  
[2] MALLO J J,RIOPEREZ J,HONRUBIAA P.The addition of Enterococcus faecium to diet improves piglet's intestinal microbiota and performance[J].Livestock Science,2010,133(1/2/3):176-178.
[3] EGLI K,LANGER C,SIEGRIST H R,et al.Community analysis of ammonia and nitrite oxidizers during start-up of nitritation reactors[J].Applied and Environmental Microbiology,2003,69(6):3213-3222.  
[4] FIFUEROLA E L M,ERIJMAN L.Bacterial taxa abundance pattern in an industrial wastewater treatment system determined by the full rRNA cycle approach[J].Environmental Microbiology,2007,9(7):1780-1789.  
[5] STAMPER D M,WALCH M,JACOBS R N.Bacterial population changes in a membrane bioreactor for graywater treatment monitored by denaturing gradient gel electrophoretic analysis of 16S rRNA gene fragments[J].Applied and Environmental Microbiology,2003,69(2):852-860.  
[6] WANG X H,WEN X H,YAN H J,et al.Bacterial community dynamics in a functionally stable pilot-scale wastewater treatment plant[J].Bioresource Technology,2011,102(3):2352-2357.  
[7] ROESCH L F,FULTHORPE R R,RIVA A,et al.Pyrosequencing enumerates and contrasts soil microbial diversity[J].The ISME Journal,2007,1(4):283-290.
[8] ZHANG X P,FU L Q,DENG B,et al.Bacillus subtilis SC02 supplementation causes alterations of the microbial diversity in grass carp water[J].World Journal of Microbiology Biotechnology,2013,29(9):1645-1653.  
[9] SCHLOSS P D,WESTCOTT S L,RYABIN T,et al.Introducing mothur:open-source,platform-independent,community-supported software for describing and comparing microbial communities[J].Applied and Environmental Microbiology,2009,75:7537-7541.
[10] PRUESSE E,QUAST C,KNITTEL K,et al.SILVA:a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB[J].Nucleic Acids Research,2007,35(21):7188-7196.  
[11] WANG Q,GARRITY G M,TIEDJE J M,et al.Nave Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J].Applied and Environmental Microbiology,2007,73(16):5261-5267.  
[12] HUSON D H,MITRA S,RUSCHEWEYH H J,et al.Integrative analysis of environmental sequences using MEGAN4[J].Genome Research,2011,21(9):1552-1560.  
[13] JOHNSON I R,BALL R O,BARACOS V E,et al.Glutamine supplementation influences immune development in the newly weaned piglet[J].Developmental and Comparative Immunology,2006,30(12):1191-1202.  
[14] CASTILLO M,MARTÍN-ORE S M,NOFRARÍAS M,et al.Changes in caecal microbiota and mucosal morphology of weaned pigs[J].Veterinary Microbiology,2007,124(3/4):239-247.
[15] SU Y,YAO W,PEREZ-GUTIERREZ O N,et al.16S ribosomal RNA-based methods to monitor changes in the hindgut bacterial community of piglets after oral administration of Lactobacillus sobrius S1[J].Anaerobe,2008,14(2):78-86.  
[16] 瞿继跃,周志洪,储国华,等.仔猪断奶腹泻及断奶前期限饲对增重的影响[J].杭州农业科技,2000(3):72-74.
[17] BUCHANAN R E,GIBBONS N E.伯杰细菌鉴定手册[M].8版.中国科学院微生物研究所,译.北京:科学出版社,1984:729-735.
[18] 左之才,刘兵,李莉,等.早期断乳应激性腹泻对仔猪肠道形态结构与肠道菌群的影响[J].中国兽医科学,2012,42(1):64-68
[19] 何明清,廖德惠,谢镜怀,等.猪不同日龄及不同肠段正常肠菌群的研究[J].畜牧兽医学报,1985,16(1):67-42.
[20] SEARS C L.A dynamic partnership:celebrating our gut flora[J].Anaerobe,2005,11(5):247-251.  
[21] USHIJIMA T,TAKAHASHI M,TATEWAKI K,et al.A selective medium for isolation and presumptive identification of the Bacteriodes fragilis group[J].Microbiology and Immunology,1983,27(12):985-993.
[22] HOOPER L V,WONG M H,THELIN A,et al.Molecular analysis of commensal host-microbial relationships in the intestine[J].Science,2001,291(5505):881-884.  
文章导航

/