高通量测序方法分析冰鲜鸡肉保鲜期间微生物菌相变化研究
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摘要
目的分析冰鲜鸡保鲜期间微生物菌相(细菌和真菌)的变化。方法采用高通量(16srDNA high-throughputsequencing)技术对冰鲜鸡保鲜期间微生物菌相多样性及丰富度进行分析。结果研究结果表明:冰鲜鸡在保鲜期间微生物菌相多样性及丰富度总体上呈先上升后下降的趋势。在保鲜初期,细菌(Cellulosimicrobium、 Vagococcus、 Serratia)的相对丰度水平均呈下降趋势;在保鲜的中后期,假单胞菌(Pseudomonas)的相对丰度急剧增加,成为优势菌。真菌主要为担子菌门(Basidiomycetes)与子囊菌门(Ascomycetes),其中子囊菌门(Ascomycetes)最丰富;保鲜期间曲霉(Aspergillus)、白蚁(Termitomyces)和镰刀菌(Fusarium)为优势菌属。结论冰鲜鸡保鲜期间Pseudomonas急剧增加,为优势腐败菌。
Objective To analyze the changes of microbial flora(bacteria and fungi) in frozen chicken during its preservation. Methods The microbial diversity and richness during the preservation of frozen chicken were analyzed through the 16 s rDNA high-throughput technology. Results The results showed that the diversity and richness of microflora of chilled chicken increased first and then decreased. At the initial stage of the preservation,the relative abundance of bacteria(Cellulosimicrobium, Vagococcus, and Serratia) was decreased. In the middle and late stage of preservation, the relative abundance of Pseudomonas increased sharply and became the dominant bacterium. The main fungi were Basidiomycetes and Ascomycetes, Ascomycetes were the most abundant; and Aspergillus, Termitomyces and Fusarium were the dominant bacteria during the preservation period. Conclusion During the preservation of chilled chicken, Pseudomonas increased sharply and became the dominant spoilage bacterium.
Objective To analyze the changes of microbial flora(bacteria and fungi) in frozen chicken during its preservation. Methods The microbial diversity and richness during the preservation of frozen chicken were analyzed through the 16 s rDNA high-throughput technology. Results The results showed that the diversity and richness of microflora of chilled chicken increased first and then decreased. At the initial stage of the preservation,the relative abundance of bacteria(Cellulosimicrobium, Vagococcus, and Serratia) was decreased. In the middle and late stage of preservation, the relative abundance of Pseudomonas increased sharply and became the dominant bacterium. The main fungi were Basidiomycetes and Ascomycetes, Ascomycetes were the most abundant; and Aspergillus, Termitomyces and Fusarium were the dominant bacteria during the preservation period. Conclusion During the preservation of chilled chicken, Pseudomonas increased sharply and became the dominant spoilage bacterium.
引文
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[8]Yang XY,Zhu LX,Zhang YM,et al.Microbial community dynamics analysis by high-throughput sequencing in chilled beef longissimus steaks packaged under modified atmospheres[J].Meat Sci,2018,141:94-102.
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[10]Soto DR,Maria DD,Dalmasso A,et al.Characterization of bacterial communities of donkey milk by high-throughput sequencing[J].Int JFood Microbiol,2017,251:67-72.
[11]Doulgeraki AI,Ercolini D,Villani F,et al.Spoilage microbiota associated to the storage of raw meat in different conditions[J].Int J Food Microbiol,2012,157(2):130-141.
[12]Higgins H,Pal C,Sulaiman IM,et al.Application of high-throughput pyrosequencing in the analysis of microbiota of food commodities procured from small and large retail outlets in a U.S.metropolitan area-Apilot study[J].Food Res Int,2017,105:29-40.
[13]Wang XH,Zhang YL,Ren HY,et al.Comparison of bacterial diversity profiles and microbial safety assessment of salami,Chinese dry-cured sausage and Chinese smoked-cured sausage by high-throughput sequencing[J].LWT,2018,90:108-115.
[14]Labadie L.Consequences of packaging on bacterial growth.Meat is an ecological niche[J].Meat Sci,1999,52(3):299-305.
[15]Zang JH,Xu YS,Xia WS,et al.Dynamics and diversity of microbial community succession during fermentation of Suan yu,a Chinese traditional fermented fish,determined by high throughput sequencing[J].Food Res Int,2018,111:565-573.
[16]江芸,高峰,徐幸莲,等.冷却猪肉不同前处理对细菌DNA提取及PCR-DGGE的影响[J].食品科学,2010,31(6):258-262.Jiang Y,Gao F,Xu XL,et al.Effects of different pretreatment of chilled pork on bacterial DNA extraction and PCR-DGGE[J].Food Sci,2010,31(6):258-262.
[17]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.Xie P,Xu MS,Yin ZP,et al.MiSeq sequencing study on the microbial community diversity during the storage of bulk sauce brine duck meat[J].Mod Food Sci Technol,2015,31(11):120-126,106.
[18]Hess M,Sczyrba X,Egan R,et al.Metagenomic discovery of biomass-degrading genes and genomes from cow rumen[J].Science,2011,331(6016):463-467.
[19]Haas BJ,Gevers D,Earl AM,et al.Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons[J].Genome Res,2011,21(3):494-504.
[20]Desantis TZ,Hugenholtz P,Larsen N,et al.Greengenes,a chimera-checked 16S rRNA gene database and workbench compatible with ARB[J].Appl Environ Microbiol,2006,72(7):5069-5072.
[21]Altschul SF,Gish W,Miller EW,et al.Basic local alignment search tool[J].J Mol Biol,1990,215(3):403-410.
[22]Urmas K,Nilsson RH,Abarenkov K,et al.Towards a unified paradigm for sequence-based identification of fungi[J].Mol Ecol,2013,22(21):5271-5277.
[23]Dorn-In S,H?lzel CS,Janke T,et al.PCR-SSCP-based reconstruction of the original fungal flora of heat-processed meat products[J].Int J Food Microbiol,2013,162(1):71-81.
[24]王玉荣,廖华,赵慧君,等.基于PCR-DGGE与高通量测序技术的恩施地区腊鱼细菌多样性评价[J].现代食品科技,2018,34(11):208-213,175.Wang YR,Liao H,Zhao HJ,et al.Evaluation of bacterial diversity in enshi region based on pcr-dgge and high-throughput sequencing technology[J].Mod Food Sci Technol,2018,34(11):208-213,175.
[25]Rong C,Ling Z,Huihui S,et al.Effect of carbonized apple branches on bacterial and fungal diversities in apple root-zone soil[J].Sci Agric Sin,2016,49(17):3414-3424.
[26]Roesch LFW,Fulthorpe RR,Riva A,et al.Pyrosequencing enumerates and contrasts soil microbial diversity[J].ISME J,2007,1(4):327-327.
[27]Zang JH,Xu YS,Xia WS,et al.Dynamics and diversity of microbial community succession during fermentation of Suan yu,a Chinese traditional fermented fish,determined by high throughput sequencing[J].Food Res Int,2018,111:565-573.
[28]戴宝玲,杨华,王佩佩,等.家禽定点屠宰场不同屠宰环节鸡肉表面真菌结构分析[J].浙江农业科学,2018,59(8):1409-1412.Dai BL,Yang H,Wang PP,et al.Analysis on the structure of chicken surface fungi at different slaughter stages in poultry slaughterhouses[J].Zhejiang Agric Sci,2018,59(8):1409-1412.
[29]Gje N,Skandamis PN,Tassou CC,et al.Meat spoilage during distribution[J].Meat Sci,2008,78(1):77-89.
[30]Gill CO,Ahvenainen R.Active packaging in practice:Meat[M].Novel Food Packaging Techniques,2003.
[31]Pettersen MK,Nissen H,Eie T,et al.Effect of packaging materials and storage conditions on bacterial growth,off-odour,pH and colour in chicken breast fillets[J].Packag Technol Sci,2010,17(3):165-174.
[32]Zhang YT,Yao YG,Gao LF,et al.Characterization of a microbial community developing during refrigerated storage of vacuum packed Yao meat,a Chinese traditional food[J].LWT,2018,(90):562-569.
[33]Lee,Weon CM,Hong HY,et al.Cellulomonas aerilata sp.nov.isolated from an air sample[J].Int J System Evolut Microbiol,2008,58(12):2925-2929.
[34]Hatayama K,Esaki K,Ide T.Cellulomonas soli sp.nov.and Cellulomonas oligotrophica sp.nov.isolated from soil[J].Int J System Evolut Microbiol,2012,63(1):60-65.
[35]Wallbanks S,Martinez-Murcia AJ,Fryer JL,et al.16S rRNA sequence determination for members of the genus Carnobacterium and related lactic acid bacteria and description of Vagococcus salmoninarum sp.nov[J].Int J System Bacteriol,1990,40(3):224-30.
[36]Carrizosa E,Benito M J,Ruiz-Moyano S,et al.Bacterial communities of fresh goat meat packaged in modified atmosphere[J].Food Microbiol,2017,65:57-63.
[37]汤飞.冷鲜鸡肉生产过程中微生物污染分析及微冻保鲜技术研究[D].合肥:合肥工业大学,2017.Tang F.Analysis of microbiological contamination and microfrozen preservation technology in the production of chilled chicken[D].Hefei:Hefei University of Technology,2017.
[2]田晓萌,李艺杰,季冬青,等.我国带鱼保鲜技术研究进展[J].保鲜与加工,2017,(4):127-132.Tian XM,Li YJ,Ji DQ,et al.Research progress on the preservation technology of hairtail fish in China[J].Preserv Process,2017,(4):127-132.
[3]许立兴,关文强,荆红彭,等.猪肉超冰温保鲜效果研究[C].第七届中国冷冻冷藏新技术新设备研讨会论文集,2015.Xu LX,Guan WQ,Jing HP,et al.Research on the effect of super cold temperature preservation of pork[C].Proceedings of the 7th China Symposium on New Technology and New Equipment for Refrigeration,2015.
[4]温冬玲,成淑君,刘悦,等.高通量测序分析不同增菌温度下冷鲜鸡肉细菌的群落多样性[J].食品科学,2018,39(24):156-161.Wen DL,Cheng SJ,Liu Y,et al.High-throughput sequencing analysis of bacterial community diversity in chilled chicken at different bacteria enrichment temperatures[J].Food Sci,2018,39(24):156-161.
[5]李忠辉,姚开,贾冬英,等.冷鲜鸡胸肉主要腐败菌的分离及低温贮藏对货架期的影响[J].食品与发酵工业,2011,37(1):167-170.Li ZH,Yao K,Jia DY,et al.Isolation of predominant spoilage bacteria in chilled chicken breast and evaluation of the shelf life at low storage temperatures[J].Food Ferment Ind,2011,37(1):167-170.
[6]刘朏.不同贮藏温度对冷鲜鸡微生物和肉品品质的影响研究[D].西安:西北农林科技大学,2015.Liu F.Effect of different storage temperatures on the microorganism and meat quality of chilled chickens[D].Xian:Northwest university of agriculture and forestry science and technology,2015.
[7]Wang TJ,Zhao L,Sun YN,et al.Changes in the microbiota of lamb packaged in a vacuum and in modified atmospheres during chilled storage analysed by high-throughput sequencing[J].Meat Sci,2016,121:253-260.
[8]Yang XY,Zhu LX,Zhang YM,et al.Microbial community dynamics analysis by high-throughput sequencing in chilled beef longissimus steaks packaged under modified atmospheres[J].Meat Sci,2018,141:94-102.
[9]孙彦雨,周光宏,徐幸莲.冰鲜鸡肉贮藏过程中微生物菌相变化分析[J].食品科学,2011.32(11):146-151.Sun YY,Zhou GH,Xu XL.Analysis of microorganism and bacterial phase changes in chilled chicken during storage[J].Food Sci,2011,32(11):146-151.
[10]Soto DR,Maria DD,Dalmasso A,et al.Characterization of bacterial communities of donkey milk by high-throughput sequencing[J].Int JFood Microbiol,2017,251:67-72.
[11]Doulgeraki AI,Ercolini D,Villani F,et al.Spoilage microbiota associated to the storage of raw meat in different conditions[J].Int J Food Microbiol,2012,157(2):130-141.
[12]Higgins H,Pal C,Sulaiman IM,et al.Application of high-throughput pyrosequencing in the analysis of microbiota of food commodities procured from small and large retail outlets in a U.S.metropolitan area-Apilot study[J].Food Res Int,2017,105:29-40.
[13]Wang XH,Zhang YL,Ren HY,et al.Comparison of bacterial diversity profiles and microbial safety assessment of salami,Chinese dry-cured sausage and Chinese smoked-cured sausage by high-throughput sequencing[J].LWT,2018,90:108-115.
[14]Labadie L.Consequences of packaging on bacterial growth.Meat is an ecological niche[J].Meat Sci,1999,52(3):299-305.
[15]Zang JH,Xu YS,Xia WS,et al.Dynamics and diversity of microbial community succession during fermentation of Suan yu,a Chinese traditional fermented fish,determined by high throughput sequencing[J].Food Res Int,2018,111:565-573.
[16]江芸,高峰,徐幸莲,等.冷却猪肉不同前处理对细菌DNA提取及PCR-DGGE的影响[J].食品科学,2010,31(6):258-262.Jiang Y,Gao F,Xu XL,et al.Effects of different pretreatment of chilled pork on bacterial DNA extraction and PCR-DGGE[J].Food Sci,2010,31(6):258-262.
[17]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.Xie P,Xu MS,Yin ZP,et al.MiSeq sequencing study on the microbial community diversity during the storage of bulk sauce brine duck meat[J].Mod Food Sci Technol,2015,31(11):120-126,106.
[18]Hess M,Sczyrba X,Egan R,et al.Metagenomic discovery of biomass-degrading genes and genomes from cow rumen[J].Science,2011,331(6016):463-467.
[19]Haas BJ,Gevers D,Earl AM,et al.Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons[J].Genome Res,2011,21(3):494-504.
[20]Desantis TZ,Hugenholtz P,Larsen N,et al.Greengenes,a chimera-checked 16S rRNA gene database and workbench compatible with ARB[J].Appl Environ Microbiol,2006,72(7):5069-5072.
[21]Altschul SF,Gish W,Miller EW,et al.Basic local alignment search tool[J].J Mol Biol,1990,215(3):403-410.
[22]Urmas K,Nilsson RH,Abarenkov K,et al.Towards a unified paradigm for sequence-based identification of fungi[J].Mol Ecol,2013,22(21):5271-5277.
[23]Dorn-In S,H?lzel CS,Janke T,et al.PCR-SSCP-based reconstruction of the original fungal flora of heat-processed meat products[J].Int J Food Microbiol,2013,162(1):71-81.
[24]王玉荣,廖华,赵慧君,等.基于PCR-DGGE与高通量测序技术的恩施地区腊鱼细菌多样性评价[J].现代食品科技,2018,34(11):208-213,175.Wang YR,Liao H,Zhao HJ,et al.Evaluation of bacterial diversity in enshi region based on pcr-dgge and high-throughput sequencing technology[J].Mod Food Sci Technol,2018,34(11):208-213,175.
[25]Rong C,Ling Z,Huihui S,et al.Effect of carbonized apple branches on bacterial and fungal diversities in apple root-zone soil[J].Sci Agric Sin,2016,49(17):3414-3424.
[26]Roesch LFW,Fulthorpe RR,Riva A,et al.Pyrosequencing enumerates and contrasts soil microbial diversity[J].ISME J,2007,1(4):327-327.
[27]Zang JH,Xu YS,Xia WS,et al.Dynamics and diversity of microbial community succession during fermentation of Suan yu,a Chinese traditional fermented fish,determined by high throughput sequencing[J].Food Res Int,2018,111:565-573.
[28]戴宝玲,杨华,王佩佩,等.家禽定点屠宰场不同屠宰环节鸡肉表面真菌结构分析[J].浙江农业科学,2018,59(8):1409-1412.Dai BL,Yang H,Wang PP,et al.Analysis on the structure of chicken surface fungi at different slaughter stages in poultry slaughterhouses[J].Zhejiang Agric Sci,2018,59(8):1409-1412.
[29]Gje N,Skandamis PN,Tassou CC,et al.Meat spoilage during distribution[J].Meat Sci,2008,78(1):77-89.
[30]Gill CO,Ahvenainen R.Active packaging in practice:Meat[M].Novel Food Packaging Techniques,2003.
[31]Pettersen MK,Nissen H,Eie T,et al.Effect of packaging materials and storage conditions on bacterial growth,off-odour,pH and colour in chicken breast fillets[J].Packag Technol Sci,2010,17(3):165-174.
[32]Zhang YT,Yao YG,Gao LF,et al.Characterization of a microbial community developing during refrigerated storage of vacuum packed Yao meat,a Chinese traditional food[J].LWT,2018,(90):562-569.
[33]Lee,Weon CM,Hong HY,et al.Cellulomonas aerilata sp.nov.isolated from an air sample[J].Int J System Evolut Microbiol,2008,58(12):2925-2929.
[34]Hatayama K,Esaki K,Ide T.Cellulomonas soli sp.nov.and Cellulomonas oligotrophica sp.nov.isolated from soil[J].Int J System Evolut Microbiol,2012,63(1):60-65.
[35]Wallbanks S,Martinez-Murcia AJ,Fryer JL,et al.16S rRNA sequence determination for members of the genus Carnobacterium and related lactic acid bacteria and description of Vagococcus salmoninarum sp.nov[J].Int J System Bacteriol,1990,40(3):224-30.
[36]Carrizosa E,Benito M J,Ruiz-Moyano S,et al.Bacterial communities of fresh goat meat packaged in modified atmosphere[J].Food Microbiol,2017,65:57-63.
[37]汤飞.冷鲜鸡肉生产过程中微生物污染分析及微冻保鲜技术研究[D].合肥:合肥工业大学,2017.Tang F.Analysis of microbiological contamination and microfrozen preservation technology in the production of chilled chicken[D].Hefei:Hefei University of Technology,2017.