基于16S rRNA高通量测序技术分析生鲜水牛乳在加工前的细菌多样性  

Analy on Bacterial Diversity of Fresh Buffalo Milk before Processing Based on 16S rRNA High-throughput Sequencing Technology

作　　者：谢芳[1] 谢华德 李孟伟[1] 唐振华[1] 杨承剑[1] XIE Fang;XIE Hua-de;LI Meng-wei;TANG Zhen-hua;YANG Cheng-jian(Guangxi Buffalo Research Institute,Chinese Academy of Agricultural Sciences,Nanning 530001)

机构地区：[1]中国农业科学院广西水牛研究所/广西水牛遗传繁育重点实验室,南宁530001

出　　处：《中国奶牛》2021年第9期52-58,共7页China Dairy Cattle

基　　金：广西自然科学基金项目(2018GXNSFBA281006、2018GXNSFAA281162);国家重点研发计划项目子课题(2018YFD0501600-02、2016YFD0500507);广西水牛遗传繁育重点实验室自主研究课题(2018-A-03-02)。

摘　　要：本研究利用16S rRNA高通量测序分析生鲜水牛乳加工前的细菌多样性。分别提取刚挤出30min内,及冷藏5h、12h及24h的水牛乳样本细菌总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进行高通量测序及BLAST比对。结果显示,在属水平,刚挤出30min~冷藏5h组中的优势菌属为金黄杆菌属(39.28%)、巨型球菌属(16.47%)、乳球菌属(9.61%),而在冷藏12~24h组中则以不动杆菌属(34.95%)、芽孢杆菌属(11.2%)、库特氏菌属(9.01%)为优势菌属。葡萄球菌属在刚挤出组中未检出,但随着冷藏时间的延长,其丰度呈上升趋势。可以得出,生鲜水牛乳从刚挤出至冷藏24h内,其微生物群落结构和组成呈动态变化,且此次测序中条件致病菌检出率较高,冷藏12h后样本组中细菌多样性显著高于其他组,由此可知生鲜水牛乳从挤出至加工前,低温贮藏时间越短越好,最好控制在5h内。To analyze the bacterial diversity of fresh buffalo milk before processing by 16S rRNA high-throughput se-quencing.Total bacterial DNA of buffalo milk samples was extracted within 30min of fresh extrusion and refrigerated for 5h,12h and 24h respectively.The 16S rDNA was amplified by PCR.The purified amplified fragments were used to construct the 16S rDNA library of the bacterial community.Results showed that,at the genus level,the dominant bacteria were Chryseobacterium(39.28%),megacoccus(16.47%)and Lactococcus(9.61%)in the group of 30 min to 5h of cold storage,while Acinetobacter(34.95%),Bacillus(11.2%)and kurtella(9.01%)were the dominant bacteria in the cold storage group of 12~24h.Staphylococcus was not detected in the extrusion group,but its abundance increased with the extension of cold storage time.Fresh buffalo milk from just out to refrigerated within 24h,the microbial community structure and composition of dynamic change,and the sequencing,conditional pathogenic bacteria detection rate is higher,refrigerated after 12h bacterial diversity in the sample group was significantly higher than that of other groups,thus the raw buffalo milk from extrusion to processing,low temperature storage time as short as possible,the best control within 5h.

关 键 词：16S rRNA高通量测序 生鲜水牛乳 冷藏 细菌多样性 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程]