基于高通量测序技术分析患乳腺炎水牛生乳中的细菌多样性  

Research of Microbial Diversity of Water Buffalo Milk During Mastitis by Using High-throughput Sequencing

作　　者：唐振华[1] 李开坤 谢芳[1] 李孟伟[1] 郭艳霞[1] 彭丽娟 彭开屏 杨承剑[1] 梁辛[1] TANG Zhenhua;LI Kaikun;XIE Fang;LI Mengwei;GUO Yanxia;PENG Lijuan;PENG Kaiping;YANG Chengjian;LIANG Xin(Guangxi Buffalo Research Institute,Chinese Academy of Agricultural Science,Guangxi Key Laboratory of Buffalo Genetics,Breeding and Reproduction,Nanning 530001,China;Guangxi Yulin Animal husbandry Station,Yulin 537000,China)

机构地区：[1]中国农业科学院广西水牛研究所农业部(广西)水牛遗传繁育重点实验室,南宁530001 [2]玉林市畜牧站,广西玉林537000

出　　处：《中国乳品工业》2020年第12期10-15,共6页China Dairy Industry

基　　金：国家重点研发计划项目(2018YFD0501600-02,2016YFD0500507);广西自然科学基金项目(2018GXNSFBA281006);广西水牛研究所基本科研业务费项目(180506,201905013)。

摘　　要：研究了尼里-拉菲水牛乳腺炎生牛乳中微生物多样性。试验选取尼里-拉菲健康奶水牛和乳腺炎奶水牛各5头作为试验动物,通过无菌操作获得奶样,测定电阻率后送回实验室测定体细胞数、提取总DNA。并对16S r RNA进行扩增,利用Illumina MiSeq测序平台进行高通量测序,进行Alpha多样性分析。结果表明:(1)乳腺炎组shannon指数均显著高于健康组,simpson指数显著低于健康组(P<0.05)。sobs、chao、ace指数差异不显著(P>0.05)。覆盖度(coverage%)所有样品均超过99%。(2)在门(Phylum)、目(Order)、属(Genus)和种(Species)水平上,占主导地位的变形菌门(proteobacteria)、β变形菌目(Betaproteobacteriales)、代尔夫特菌属(Delftia)和代尔夫特菌(Delftiatsuruhatensis)乳腺炎组(R)显著高于健康组(C)。(3)属(Genus)水平金黄杆菌(Chryseobacterium)、棒状杆菌属(Corynebacterium1)乳腺炎组显著高于健康组(P<0.05),且链球菌属(Streptococcus)、葡萄球菌属(Staphylococcus)仅在乳腺炎组(R)中检测到。因此认为乳腺炎会对奶水牛乳中细菌多样性造成改变,显著降低奶水牛乳中细菌主要门(Phylum)、目(Order)、属(Genus)和种(Species)丰度,且金黄杆菌(Chryseobacterium)、棒状杆菌属(Corynebacterium1)、链球菌属(Streptococcus)和葡萄球菌属(Staphylococcus)与奶水牛乳腺炎发生相关。The aim of this study was to investigate the microbiota of water buffalo milk during mastitis.10 sample were included in the experimental design,5 milk sample collected from buffalo with clinical mastitis and 5 samples from healthy.Somatic Cell Count(SCC),electrical resistance and DNA extraction were performed on all the samples,follow by high-throughput sequencing of the 16S rRNA gene.The results showed that:(1)The shannon index of Group mastitis was higher than Group healthy significantly(P<0.05).There is no difference on sobs,chao,ace,and the coverage of all samples is greater than 99%.(2)The relative abundance of proteobacteria、Betaproteobacteriales、Delftia and Delftiatsuruhatensis in Group mastitis were greater than Group healthy significantly(P<0.05).(3)The relative abundance of Chryseobacterium、Corynebacterium1 of Group masitis were greater than Group healthy(P<0.05),while Streptococcus、Staphylococcus were found in Group masitis only.Conclusion:The mastitis improved the microbial diversity of buffalo milk and no effects on The relative abundance of microbiota,and changed the microbial community structure on the level of order and genus.

关 键 词：乳腺炎 奶水牛 牛乳 微生物多样性 

分 类 号：S858.23[农业科学—临床兽医学] Q93-331[农业科学—兽医学]