基于16S rDNA高通量测序分析水牛初乳与常乳中细菌多样性

Analysis of Bacterial Diversity in Colostrum and Normal Milk of Buffalo Based on 16S rDNA High-throughput Sequencing

目的:利用16S rDNA高通量测序研究广西水牛研究所种畜厂三品杂水牛初乳与常乳的细菌多样性。方法:提取当天采集的生水牛乳样本的细菌总DNA,PCR扩增其16S rDNA,利用纯化后的扩增片段构建其菌群的16S rDNA文库,采用Miseq PE300平台进行高通量测序以及BLAST比对。结果:初乳组样本共得到19个门,292个属和437个种。而常乳两组样本共得到7个门,203个细菌属和330个细菌种,初乳组的细菌多样性高于常乳组。初乳组中的优势菌属为金黄杆菌属(Chryseobacterium)、不动杆菌属(Acinetobacter)、假单胞菌属(Pseusomonas),而常乳组中的优势菌则为金黄杆菌属(Chryseobacterium)、乳球菌属(Lactococcus)、肠球菌属(Enterococcus)。样本层级聚类显示:初乳组与常乳组组间群落差异不显著。PCA与PLS-DA分析表明:两组样本组间菌群结构差异显著。结论:三品杂水牛初乳与常乳均呈现较丰富的多样性,常乳组中细菌多样性显著高于初乳组。

Objective:16 S rDNA high-throughput sequencing was used to study the bacterial diversity of colostrum and normal milk of the three-breed crossbreds.Methods:The total bacterial DNA was extracted from the raw milk samples collected on the same day,the 16 S rDNA of the bacteria was amplified by PCR,and the 16 S rDNA library of the bacteria group was constructed by the purified amplified fragment.Miseq PE300 platform was used for sequencing and blast comparison.Results:There were 19 phyla,292 genera and 437 species in colostrum group.7 phyla,203 bacterial genera and 330 bacterial species were obtained from the two groups of the normal milk group.The bacterial diversity of colostrum group was higher than that of the normal milk group.Chryseobacterium,Acinetobacter,and Pseudomonas were the dominant bacteria in the colostrum group,while Chryseobacterium,Lactococcus,and Enterococcus were the dominant bacteria in the normal group.Cluster analysis showed that there was no significant difference between the two groups.PCA and PLS-DA analysis showed that there were significant differences in bacterial flora structure between the two groups.Conclusion:The diversity of colostrum and normal milk of the three-breed crossbreds is rich,and the bacterial diversity in normal milk group is significantly higher than that in colostrum group.