基于16S rDNA高通量测序技术的酱卤熟肉制品细菌多样性研究
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摘要
目的探讨不同包装类型、不同品种肉类来源酱卤熟肉制品中的菌群组成及丰度。方法提取散装及预包装酱卤熟肉制品中的细菌基因组DNA,应用高通量测序技术对样品中细菌16S rDNA V4变异区域进行测序,分析样品中微生物群体分布和丰度、以及不同样品中的菌群差异。结果不同酱卤熟肉样品菌群结构各不相同且差异较大,其中弧菌属丰度均较高,其次为不动杆菌属、希瓦菌属及嗜冷杆菌属,部分样品尚存在葡萄球菌属、沙门菌属及李斯特菌属等食源性致病菌成分。Alpha多样性结果显示,所有样品菌群丰富度及均匀度均较高,散装样品菌群丰富度明显高于预包装样品;Beta多样性分析发现散装与预包装样品组间菌群差异有统计学意义,散装样品中菌群OTU数明显多于预包装样品。结论高通量深度测序技术可快速精确地掌握熟肉制品中微生物菌群多样性,弧菌菌属可能与酱卤类熟肉制品的变质密切相关,散装类酱卤熟肉制品微生物污染状况比预包装样品普遍要高,存在引发食源性疾病的风险。
Objective To analyze the bacterial community diversity of braised meats from different sources or in different packages. Methods The bacterial DNA in the bulk and pre-packaged braised meat products was extracted, and the 16 S rDNA V4 variant region of the bacteria was sequenced by high-throughput sequencing technology. The distribution and abundance of microbial community as well as the species difference of bacteria were obtained. Results Bacterial community diversity varied in different braised meat samples. The abundance of Vibrio in all samples was higher, followed by Acinetobacter, Shewanella, and Psychrotrophs. Some samples still contained food-borne pathogenic bacteria such as genus Staphylococcus, Salmonella, and Listeria. Alpha diversity results showed that richness and evenness of the microbiota in all samples were high. Bacterial richness of bulk samples was significantly higher than that of prepackaged samples. Beta diversity analysis revealed a statistical significance on the difference between the bulk and prepackaged sample groups, and the number of OTUs in the bulk sample was significantly greater than the prepackaged sample. Conclusion High-throughput sequencing technology can be used to obtain the diversity of microbial flora in braised meat products quickly and accurately. Vibrio spp may be closely related to the deterioration of braised meat products. The microbiological contamination of bulk braised meat products is generally higher than that of prepackaged samples, and it may lead to a risk of foodborne disease.
Objective To analyze the bacterial community diversity of braised meats from different sources or in different packages. Methods The bacterial DNA in the bulk and pre-packaged braised meat products was extracted, and the 16 S rDNA V4 variant region of the bacteria was sequenced by high-throughput sequencing technology. The distribution and abundance of microbial community as well as the species difference of bacteria were obtained. Results Bacterial community diversity varied in different braised meat samples. The abundance of Vibrio in all samples was higher, followed by Acinetobacter, Shewanella, and Psychrotrophs. Some samples still contained food-borne pathogenic bacteria such as genus Staphylococcus, Salmonella, and Listeria. Alpha diversity results showed that richness and evenness of the microbiota in all samples were high. Bacterial richness of bulk samples was significantly higher than that of prepackaged samples. Beta diversity analysis revealed a statistical significance on the difference between the bulk and prepackaged sample groups, and the number of OTUs in the bulk sample was significantly greater than the prepackaged sample. Conclusion High-throughput sequencing technology can be used to obtain the diversity of microbial flora in braised meat products quickly and accurately. Vibrio spp may be closely related to the deterioration of braised meat products. The microbiological contamination of bulk braised meat products is generally higher than that of prepackaged samples, and it may lead to a risk of foodborne disease.
引文
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[2] 杨君娜.传统酱卤熟肉制品色泽固化技术研究[D].天津:天津商业大学,2009.
[3] 吴平芳,贺连华,石晓路,等.深圳市熟肉制品致病菌监测结果分析[J].中国卫生检验杂志,2012,22(8):1916-1917.
[4] 姚笛,夏秀芳,王颖,等.低温肉制品中微生物的危害及控制[J].肉类研究,2009(10):44-47.
[5] 刘新武,于艳妮,滕海风,等.16SrDNA序列在细菌鉴定和检测分析中的应用价值[J].医学检验与临床,2017,28(7):30-32.
[6] 刘文强,贾玉萍,赵宏坤,等.16S rRNA在细菌分类鉴定研究中的应用[J].动物医学进展,2006,27(11):15-18.
[7] Christensen JE,Stencil JA,Reed KD.Rapid identification of bacteria from positive blood cultures by terminal restriction fragment length polymorphism profile analysis of the 16S rRNA gene[J].J Clin Microbiol,2003,41(8):3790-3800.
[8] Li R,Zhu H,Ruan J,et al.De novo assembly of human genomes with massively parallel short read sequencing[J].Genome Res,2010,20(2):265-272.
[9] 秦楠,栗东芳,杨瑞馥.高通量测序技术及其在微生物学研究中的应用[J].微生物学报,2011,51(4):445-457.
[10] Bokulich NA,Subramanian S,Faith JJ,et al.Quality-filtering vastly improves diversity estimates from illumina amplicon sequencing[J].Nat Methods,2013,10(1):57-59.
[11] Edgar RC.Search and clustering orders of magnitude faster than BLAST[J].Bioinformatics,2010,26(19):2460-2461.
[12] Kuczynski J,Stombaugh J,Walters WA,et al.Using QIIME to analyze 16S rRNA gene sequences from microbial communities[J].Curr Protoc Microbiol,2012,doi:10.1002/9780471729259.mc01e05s27.
[13] Gray AN,Koo BM,Shiver AL,et al.High-throughput bacterial functional genomics in the sequencing era[J].Curr Opin Microbiol,2015,27:86-95.
[14] Valeria D'Argenio.The high-throughput analyses era:Are we ready for the data struggle?[J].High Throughput,2018,7(1):E8.
[15] Sogin ML,Morrison HG,Huber JA,et al.Microbial diversity in the deep sea and the underexplored “Rare Biosphere” [J].Proc Natl Acad Sci USA,2006,103(32):12115-12120.