气调包装的调理啤酒鲈鱼片在微冻贮藏过程中的微生物群落多样性分析
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摘要
为揭示调理啤酒鲈鱼片气调包装和微冻贮藏过程中的微生物变化规律及其腐败本质,为优化产品工艺和推广产品提供理论依据,采用Illumina MiSeq测序技术解析调理啤酒鲈鱼制品气调包装和-3℃贮藏过程中微生物群落多样性,并比较其与真空包装4℃贮藏鲈鱼片的微生物群落多样性的区别。结果表明:鲈鱼制品在贮藏过程中的优势微生物为厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes);真空包装的未调味鲈鱼片在4℃贮藏过程中微生物多样性比在-3℃贮藏的气调包装调理啤酒鲈鱼丰富,且在贮藏第4天时,开始出现了索丝菌属(Brochothrix)等常见的腐败微生物,到第10天时,索丝菌属、假单胞菌属(Pseudomonas)丰度迅速增大,出现了希瓦氏菌属(Shewanella),造成鲈鱼片彻底腐败变质;气调包装的调理啤酒鲈鱼片在第20天时才开始出现假单胞菌属和希瓦氏菌属等腐败菌,之后在CO_2的抑制作用下,腐败菌丰度逐渐减小,直到货架期终点时,假单胞菌才迅速增长,且出现了嗜冷杆菌属(Psychrobacte),造成了鲈鱼制品的彻底腐败变质。这说明气调包装联合啤酒调理的加工方式及-3℃贮藏可以有效延长产品的货架期。
This study aimed to reveal changes in the microbial communities in prepared sea bass ?llet with beer and the essence of spoilage during partially frozen storage under modi?ed atmosphere packaging for the purpose of providing a theoretical basis for optimizing processing conditions and promoting the product. Illumina MiSeq sequencing technology was used in this study to analyze the diversity of the microbial communities during storage at -3 ℃ under modified atmosphere packaging in comparison with storage at 4 ℃ under vacuum packaging. The results showed that the dominant microorganisms of prepared sea bass fillets during storage were Firmicutes, Proteobacteria and Bacteroidetes. Vacuumpackaged unseasoned sea bass ?llets stored at 4 ℃ showed more abundant microbial diversity than modi?ed-atmosphere packaged prepared sea bass fillets with beer stored at -3 ℃. On the 4~(th) day of storage, the common spoilage bacterium Brochothrix began to be detected in the unseasoned sea bass fillets, on the 10~(th) day, the abundance of Brochothrix and Pseudomonas rapidly increased, and even Shewanella began to appear, causing complete spoilage of sea bass ?llets. The spoilage bacteria Pseudomonas and Shewanella began to appear in modi?ed-atmosphere packaged sea bass ?llets with beer on the 20~(th) day. Afterwards, due to inhibition by CO_2, the abundance of spoilage bacteria gradually decreased, Pseudomonas grew rapidly towards the end of the shelf life, and Psychrobacte appeared at the end, causing complete spoilage of sea bass?llets. Therefore, the combined application of seasoning with beer, modi?ed atmosphere packaging and superchilled storage can effectively extend the shelf life of sea bass ?llets.
This study aimed to reveal changes in the microbial communities in prepared sea bass ?llet with beer and the essence of spoilage during partially frozen storage under modi?ed atmosphere packaging for the purpose of providing a theoretical basis for optimizing processing conditions and promoting the product. Illumina MiSeq sequencing technology was used in this study to analyze the diversity of the microbial communities during storage at -3 ℃ under modified atmosphere packaging in comparison with storage at 4 ℃ under vacuum packaging. The results showed that the dominant microorganisms of prepared sea bass fillets during storage were Firmicutes, Proteobacteria and Bacteroidetes. Vacuumpackaged unseasoned sea bass ?llets stored at 4 ℃ showed more abundant microbial diversity than modi?ed-atmosphere packaged prepared sea bass fillets with beer stored at -3 ℃. On the 4~(th) day of storage, the common spoilage bacterium Brochothrix began to be detected in the unseasoned sea bass fillets, on the 10~(th) day, the abundance of Brochothrix and Pseudomonas rapidly increased, and even Shewanella began to appear, causing complete spoilage of sea bass ?llets. The spoilage bacteria Pseudomonas and Shewanella began to appear in modi?ed-atmosphere packaged sea bass ?llets with beer on the 20~(th) day. Afterwards, due to inhibition by CO_2, the abundance of spoilage bacteria gradually decreased, Pseudomonas grew rapidly towards the end of the shelf life, and Psychrobacte appeared at the end, causing complete spoilage of sea bass?llets. Therefore, the combined application of seasoning with beer, modi?ed atmosphere packaging and superchilled storage can effectively extend the shelf life of sea bass ?llets.
引文
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[19]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.DOI:10.1038/nmeth.2604.
[20]熊振海,马晨晨.冷却牛肉货架期及微生物多样性分析[J].食品安全质量检测学报,2014,5(7):2109-2113.
[21]谢萍,徐明生,尹忠平,等.MiSeq测序研究散装酱卤鸭肉贮藏期间微生物群落多样性[J].现代食品科技,2015,31(11):120-126;106.DOI:10.13982/j.mfst.1673-9078.2015.11.020.
[22]梁荣蓉.生鲜鸡肉调理制品菌群结构分析和货架期预测模型的研究[D].泰安:山东农业大学,2010:40-56.
[23]范晓攀,王娉,陈颖,等.肉类调理食品中细菌多样性的分析[J].现代食品科技,2017,33(1):237-242.DOI:10.13982/j.mfst.1673-9078.2017.1.036.
[24]GILL C O,DUSSAULT F,HOLLEY R A,et al.Evaluation of the hygienic performances of the processes for cleaning,dressing and cooling pig carcasses at eight packing plants[J].International Journal of Food Microbiology,2000,58(1):65-72.DOI:10.1016/S0168-1605(00)00294-4.
[25]蔡秋杏,李来好,陈胜军,等.液熏罗非鱼片加工过程中微生物群落的PCR-DGGE分析[J].食品科学,2009,30(23):35-40.
[26]胡萍,周光宏,徐幸莲,等.应用PCR-DGGE指纹技术研究真空包装火腿切片贮藏过程中的微生物动态变化[J].南京农业大学学报,2009,32(2):137-140.
[27]HOVDA M B,LUNESTAD B T,SIVERTSVIK M,et a1.Characterisation of the bacterial flora of modified atmosphere packaged farmed Atlantic cod(Gadus morhua)by PCR-DGGE of conserved16S rRNA gene regions[J].Internal Journal of Food Microbiology,2007,117(1):68-75.DOI:10.1016/j.ijfoodmicro.2007.02.022.
[28]TORRIERI E,CAVELLA S,VILLANI F,et al.Influence of modified atmosphere packaging on the chilled shelf life of gutted farmed bass(Dicentrarchus labrax)[J].Journal of Food Engineering,2006,77(4):1078-1086.DOI.10.1016/j.jfoodeng.2005.08.038.
[29]李蕾蕾,王素英.南美白对虾在低温贮藏过程中的品质及菌相变化分析[J].食品科学,2014,35(18):246-251.DOI:10.7506/spkx1002-6630-201418047.
[30]蓝蔚青,张皖君,谢晶,等.气调包装对鲳鱼冷藏期间主要微生物多样性的影响[J].中国食品学报,2017,17(2):152-159.DOI:10.16429/j.1009-7848.2017.02.021.
[2]吴燕燕,朱小静,李来好,等.养殖日本真鲈和大口黑鲈原料特性比较[J].海洋渔业,2016,38(5):507-515.DOI:10.13233/j.cnki.mar.fish.2016.05.006.
[3]李颖畅,王亚丽,齐凤元,等.紫菜多糖提取物对冷藏鲈鱼品质的影响[J].食品工业科技,2014,35(22):336-339;348.DOI:10.13386/j.issn1002-0306.2014.22.065.
[4]魏涯,钱茜茜,吴燕燕,等.栅栏技术在淡腌半干鲈鱼加工工艺中的应用[J].南方水产科学,2017,13(2):109-120.DOI:10.3969/j.issn.2095-0780.2017.02.014.
[5]PARLAPANI F F,MALLOUCHOS A,HAROUTOUNIAN S A,et al.Microbiological spoilage and investigation of volatile profile during storage of sea bream fillets under various conditions[J].International Journal of Food Microbiology,2014,189:153-163.DOI:10.1016/j.ijfoodmicro.2014.08.006.
[6]HANDELSMAN J.Metagenomics:application of genomics to uncultured microorganisms[J].Microbiology and Molecular Biology Reviews,2004,68(4):669-685.DOI:10.1128/MMBR.68.4.669-685.2004.
[7]李建柱,侯杰,张鹏飞,等.鱼菜共生模式中不同鱼类肠道微生物群落结构的比较[J].南方水产科学,2016,12(6):42-50.DOI:10.3969/j.issn.2095-0780.2016.06.006.
[8]张岩,吴燕燕,李来好,等.16S rDNA序列分析鉴定一株合浦珠母贝共附生乳酸菌[J].南方水产科学,2012,8(6):9-15.DOI:10.3969/j.issn.2095-0780.2012.06.002.
[9]PO?KA J,REBECCHI A,PISACANE V,et al.Bacterial diversity in typical Italian salami at different ripening stages as revealed by high-throughput sequencing of 16S rRNA amplicons[J].Food Microbiology,2015,46:342-356.DOI:10.1016/j.fm.2014.08.023.
[10]WILLIAMS S T,FOSTER P G,LITTLEWOOD D T J.The complete mitochondrial genome of a turbinid vetigastropod from MiSeq Illumina sequencing of genomic DNA and steps towards a resolved gastropod phylogeny[J].Gene,2014,533(1):38-47.DOI:10.1016/j.gene.2013.10.005.
[11]YOUSSEF N,SHEIK C S,KRUMHOLZ L R,et al.Comparison of species richness estimates obtained using nearly complete fragments and simulated pyrosequencing-generated fragments in 16S rRNAgene-based environmental surveys[J].Applied and Environmental Microbiology,2009,75(16):5227-5236.DOI:10.1128/AEM.00592-09.
[12]吴燕燕,钱茜茜,李来好,等.基于Illumina MiSeq技术分析腌干鱼加工过程中微生物群落多样性[J].食品科学,2017,38(12):1-8.DOI:10.7506/spkx1002-6630-201712001.
[13]聂志强,韩玥,郑宇,等.宏基因组学技术分析传统食醋发酵过程微生物多样性[J].食品科学,2013,34(15):198-203.DOI:10.7506/spkx1002-6630-201315041.
[14]吴燕燕,朱小静,李来好,等.比较调理啤酒鲈鱼片在不同贮藏条件下的品质变化[J].食品科学,2018,39(11):214-220.DOI:10.7506/spkx1002-6630-201811034.
[15]HUSE S M,DETHLEFSEN L,HUBER J A,et al.Exploring microbial diversity and taxonomy using SSU rRNA hypervariable tag sequencing[J].PLoS Genetics,2008,4(11):1-10.DOI:10.1371/journal.pgen.1000255.
[16]MAGOC T,SALZBERG S L.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2963.DOI:10.1093/bioinformatics/btr507.
[17]BOKULICH N A,SUBRAMANIAN S,FAITH J J,et al.Qualityfiltering vastly improves diversity estimates from Illumina amplicon sequencing[J].Nature Methods,2012,10(1):57-59.DOI:10.1038/nmeth.2276.
[18]BULGARELLI D,GARRIDO-OTER R,MüNCH P C,et al.Structure and function of the bacterial root microbiota in wild and domesticated barley[J].Cell Host&Microbe,2015,17(3):392-403.DOI:10.1016/j.chom.2015.01.011.
[19]EDGAR R C.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.DOI:10.1038/nmeth.2604.
[20]熊振海,马晨晨.冷却牛肉货架期及微生物多样性分析[J].食品安全质量检测学报,2014,5(7):2109-2113.
[21]谢萍,徐明生,尹忠平,等.MiSeq测序研究散装酱卤鸭肉贮藏期间微生物群落多样性[J].现代食品科技,2015,31(11):120-126;106.DOI:10.13982/j.mfst.1673-9078.2015.11.020.
[22]梁荣蓉.生鲜鸡肉调理制品菌群结构分析和货架期预测模型的研究[D].泰安:山东农业大学,2010:40-56.
[23]范晓攀,王娉,陈颖,等.肉类调理食品中细菌多样性的分析[J].现代食品科技,2017,33(1):237-242.DOI:10.13982/j.mfst.1673-9078.2017.1.036.
[24]GILL C O,DUSSAULT F,HOLLEY R A,et al.Evaluation of the hygienic performances of the processes for cleaning,dressing and cooling pig carcasses at eight packing plants[J].International Journal of Food Microbiology,2000,58(1):65-72.DOI:10.1016/S0168-1605(00)00294-4.
[25]蔡秋杏,李来好,陈胜军,等.液熏罗非鱼片加工过程中微生物群落的PCR-DGGE分析[J].食品科学,2009,30(23):35-40.
[26]胡萍,周光宏,徐幸莲,等.应用PCR-DGGE指纹技术研究真空包装火腿切片贮藏过程中的微生物动态变化[J].南京农业大学学报,2009,32(2):137-140.
[27]HOVDA M B,LUNESTAD B T,SIVERTSVIK M,et a1.Characterisation of the bacterial flora of modified atmosphere packaged farmed Atlantic cod(Gadus morhua)by PCR-DGGE of conserved16S rRNA gene regions[J].Internal Journal of Food Microbiology,2007,117(1):68-75.DOI:10.1016/j.ijfoodmicro.2007.02.022.
[28]TORRIERI E,CAVELLA S,VILLANI F,et al.Influence of modified atmosphere packaging on the chilled shelf life of gutted farmed bass(Dicentrarchus labrax)[J].Journal of Food Engineering,2006,77(4):1078-1086.DOI.10.1016/j.jfoodeng.2005.08.038.
[29]李蕾蕾,王素英.南美白对虾在低温贮藏过程中的品质及菌相变化分析[J].食品科学,2014,35(18):246-251.DOI:10.7506/spkx1002-6630-201418047.
[30]蓝蔚青,张皖君,谢晶,等.气调包装对鲳鱼冷藏期间主要微生物多样性的影响[J].中国食品学报,2017,17(2):152-159.DOI:10.16429/j.1009-7848.2017.02.021.