肉鸭养殖过程中发酵床垫料菌群结构变化的研究  

Succession of bacterial community in duck bio-bed over time

作　　者：章小婷[1] 郑卫江[1] 林勇[1] 袁婧[1] 姚文[1] 

机构地区：[1]南京农业大学消化道微生物研究室,江苏南京210095

出　　处：《微生物学通报》2015年第7期1263-1270,共8页Microbiology China

基　　金：国家自然科学基金项目(No.31372321);国家973计划项目(No.2013CB127201)

摘　　要：【目的】探究发酵床使用时间和肉鸭粪便微生物对发酵床垫料菌群结构、总菌和大肠杆菌数量的影响。【方法】采集江苏某肉鸭发酵床养殖场内刚制作完成的发酵床垫料样品,及其饲养4批次、8批次后的垫料样品,同时采集各批次34日龄肉鸭粪便样品,采用变性梯度凝胶电泳技术(Denaturing gradient gel electrophoresis,DGGE)、16S r RNA基因序列分析和实时荧光定量PCR(Quantitative real-time PCR)技术对发酵床使用过程中垫料菌群结构进行定性和定量研究。【结果】0批次(D0)与4批次(D4)、8批次(D8)垫料菌群相似性分别为68.81%、70.82%,而4批次与8批次间垫料菌群的相似性则达81.93%,显著高于D4、D8与D0间相似性(P<0.05)。条带6、8(最相似菌分别为Leqionella tunisiensis、Pedobacte bauzanensis)在3个时间点垫料菌群中均表现优势,且含量较为稳定;条带10(最相似菌为Rummeliibacillus suwonesis)仅在2个重复使用垫料菌群中表现优势;条带12、13(最相似菌分别是Psychrobacter sp.PRwf-1、Iamia majanohamensis)共同存在于垫料样和粪便样。肉鸭粪便中大肠杆菌的数量显著高于4批次、8批次垫料中的数量(P<0.05),与0批次垫料间差异不显著(P>0.05)。【结论】使用时间和肉鸭粪便微生物共同影响了发酵床垫料菌群结构和数量,菌群结构随使用时间的延长而趋于稳定。[Objective] This study aimed to character the succession of bacterial community in the litter during the fermentation of duck bio-bed. [Methods] The experiment was conducted in one meat duck bio-bed farm of Jiangsu province. The new litter （DO）, used litter from 4th （D4）, 8th （D8） meat duck flock and feces from duck at 34 d age were sampled. Denaturing gradient gel electrophoresis （DGGE）, 16S rRNA gene sequencing and quantitative real-time PCR were used to monitor the composition changes of microbiota in the bio-bed. [Results] DGGE profiles showed that, the similarity between D4 and D8 was 81.93%, which was significantly higher than the similarity between D4, D8 and DO （68.81% and 70.82%, respectively） （P〈0.05）. Bands 6 and 8 （the closest relatives： Leqionella tunisiensis and Pedobacter bauzanensis, respectively） were dominant in all litter samples. Band 10 （the closest relative： Rummeliibacillus suwonesis） was only dominant in used litters. Bands 12 and 13 （the closest relatives： Psychrobacter sp. PRwf-1, Iamia majanohamensis, respectively） commonly presented in all litters and feces. Real-time PCR results showed that the number ofEscherichia coli in duck feces was significantly higher than that in D4 and D8 litters （P〈0.05）, while no significant difference was observed between duck feces and DO litter （P〉0.05）. [Conclusion] Both the application time ofbio-bed and duck feces origin microorganisms can affect the bacterial community and the number of E. coli in the bio-bed, and the bacterial community structure tends to be stable in line with the application time of the bio-bed.

关 键 词：发酵床 肉鸭 细菌种群 变性梯度凝胶电泳技术 实时荧光定量PCR 

分 类 号：S834[农业科学—畜牧学]