运用454焦磷酸测序技术对断奶前后仔猪肠道菌群的分析  

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

作　　者：王一冰[1] 张小平[1] 黄怡[2] 李卫芬[1] 

机构地区：[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

出　　处：《动物营养学报》2013年第10期2440-2446,共7页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：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)

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。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 postweaning 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.

关 键 词：断奶仔猪 肠道菌群 454焦磷酸测序 

分 类 号：S828[农业科学—畜牧学]