基于高通量测序分析不同保鲜冰处理对鲈鱼菌群组成与代谢功能的影响
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摘要
目的:分析不同保鲜冰处理下鲈鱼微生物菌群变化及其与品质指标的相关性。方法:分别采用流化冰(slurry ice,SI)、酸性电解水冰(acidic electrolyzed water ice,AEWI)和碎冰(crushed ice,CI)对鲈鱼进行处理,测定鲈鱼的总挥发性盐基氮(total volatile basic nitrogen,TVB-N)含量与K值,并利用高通量测序技术对样品菌群结构和代谢功能进行比较分析。结果:SI和AEWI对鲈鱼均有一定的保鲜作用,其中SI抑制鲈鱼蛋白质和核苷酸降解速度的效果最显著。在冰藏前期(0～3 d),AEWI与CI组样品中芽孢杆菌属(Bacillus)、乳球菌属(Lactococcus)、大洋芽孢杆菌属(Oceanobacillus)相对丰度明显高于SI组。冰藏中、后期(12～21 d),SI组样品中假单胞菌属(Pseudomonas)、希瓦氏菌属(Shewanella)、摩替亚氏菌属(Moritella)、埃希氏杆菌属(Escherichia)和别弧菌属(Aliivibrio)相对丰度明显增加,AEWI组样品假单胞菌属(Pseudomonas)和不动杆菌属(Acinetobacter)相对丰度所占比例逐渐增大,CI组中不动杆菌属在21 d时达27.77%,成为造成鲈鱼贮藏中、后期腐败的主要菌属。主成分分析表明,冰藏方式及贮藏时间对鲈鱼菌群结构组成有较大影响。鲈鱼菌群中与TVB-N含量、K值呈正相关的有假单胞菌属、不动杆菌属、嗜冷杆菌属(Psychrobacter)和希瓦氏菌属,它们可能是鲈鱼冰藏期间的优势菌。CI和AEWI组鲈鱼细菌中参与氨基酸、脂质和碳水化合物代谢的相关基因相对丰度高于同一时期的SI组,这在一定程度上从细菌代谢水平解释了SI能延缓鲈鱼腐败的原因,为后期进一步从微生物抑制机制上优化鲈鱼贮藏保鲜技术提供了理论参考。
Objective: To evaluate variations in the bacterial community composition of Lateolabrax japonicas during storage with different ice treatments and to determine its correlation with quality attributes. Methods: The fish were treated with slurry ice(SI), acidic electrolyzed water ice(AEWI) or crushed ice(CI). Total volatile basic nitrogen(TVB-N) content and K values(freshness index) were measured during ice storage. The structure and metabolic function of the bacterial community were also analyzed by using high-throughput sequencing. Results: SI and AEWI could maintain the quality of fish with the former being more effective in inhibiting the degradation rate of protein and nucleotide. The relative abundance of Bacillus,Lactococcus and Oceanobacillus in the CI and AEWI groups were much higher than in the SI group at the early stage of storage(0–3 d). During the middle to late periods of storage(12–21 d), the relative abundance of Pseudomonas, Shewanella,Moritella, Escherichia and Aliivibrio in the SI group increased rapidly; in the AEWI group, Pseudomonas and Acinetobacter gradually became dominant; the relative abundance of Acinetobacter as the dominant spoilage bacteria was up to 27.77%in the CI group on the 21 st day. The results of principal component analysis(PCA) showed that ice storage and storage time had a significant influence on the bacterial community composition of Lateolabrax japonicus. Pseudomonas, Acinetobacter,Psychrobacter and Shewanella were positively correlated with TVB-N contents and K values, which may be the specific spoilage organisms of Lateolabrax japonicas during ice storage. The relative abundance of genes related to amino acid, lipid and carbohydrate metabolism in the AEWI and CI group were greatly higher than in the SI group. This may partially explain why SI could delay spoilage at the level of bacterial metabolism. This study provides a theoretical basis for optimizing the preservation and storage of Lateolabrax japonicas by microbial inhibition.
Objective: To evaluate variations in the bacterial community composition of Lateolabrax japonicas during storage with different ice treatments and to determine its correlation with quality attributes. Methods: The fish were treated with slurry ice(SI), acidic electrolyzed water ice(AEWI) or crushed ice(CI). Total volatile basic nitrogen(TVB-N) content and K values(freshness index) were measured during ice storage. The structure and metabolic function of the bacterial community were also analyzed by using high-throughput sequencing. Results: SI and AEWI could maintain the quality of fish with the former being more effective in inhibiting the degradation rate of protein and nucleotide. The relative abundance of Bacillus,Lactococcus and Oceanobacillus in the CI and AEWI groups were much higher than in the SI group at the early stage of storage(0–3 d). During the middle to late periods of storage(12–21 d), the relative abundance of Pseudomonas, Shewanella,Moritella, Escherichia and Aliivibrio in the SI group increased rapidly; in the AEWI group, Pseudomonas and Acinetobacter gradually became dominant; the relative abundance of Acinetobacter as the dominant spoilage bacteria was up to 27.77%in the CI group on the 21 st day. The results of principal component analysis(PCA) showed that ice storage and storage time had a significant influence on the bacterial community composition of Lateolabrax japonicus. Pseudomonas, Acinetobacter,Psychrobacter and Shewanella were positively correlated with TVB-N contents and K values, which may be the specific spoilage organisms of Lateolabrax japonicas during ice storage. The relative abundance of genes related to amino acid, lipid and carbohydrate metabolism in the AEWI and CI group were greatly higher than in the SI group. This may partially explain why SI could delay spoilage at the level of bacterial metabolism. This study provides a theoretical basis for optimizing the preservation and storage of Lateolabrax japonicas by microbial inhibition.
引文
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[12]XIA X L,ZHANG Q W,ZHANG B,et al.Insights into the biogenic amine metabolic landscape during industrial semidry Chinese rice wine fermentation[J].Journal of Agricultural and Food Chemistry,2016,64(39):7385-7393.DOI:10.1021/acs.jafc.6b01523.
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[15]程三红,汤海青,欧昌荣,等.鲐鱼和大黄鱼冷藏期间体表细菌群落组成和代谢功能的比较分析[J].食品科学,2018,39(19):218-225.DOI:10.7506/spkx1002-6630-201819034.
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[17]于美娟,谭欢,马美湖,等.传统固态发酵鱼中细菌群落多样性与品质特征分析[J].食品科学,2017,38(8):86-95.DOI:10.7506/spkx1002-6630-201708015.
[18]农业部.水产品中挥发性盐基氮的测定:SC/T 3032-2007[S].北京:中国标准出版社,2007:3.
[19]杨文鸽,薛长湖,徐大伦,等.大黄鱼冰藏期间ATP关联物含量变化及其鲜度评价[J].农业工程学报,2007,23(6):217-222.
[20]温崇庆,何瑶瑶,薛明,等.高通量测序分析DNA提取引起的对虾肠道菌群结构偏差[J].微生物学报,2016,56(1):130-142.
[21]CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QIIMEallows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.DOI:10.1038/nmeth.f.303.
[22]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.
[23]CAI L Y,WU X S,LI X X,et al.Effects of different freezing treatments on physicochemical responses and microbial characteristics of Japanese sea bass(Lateolabrax japonicas)fillets during refrigerated storage[J].LWT-Food Science and Technology,2014,15(1):122-129.DOI:10.1016/j.lwt.2014.04.062.
[24]LI Q,ZHANG L T,LU H,et al.Comparison of postmortem changes in ATP-related compounds,protein degradation and endogenous enzyme activity of white muscle and dark muscle from common carp(Cyprinus carpio)stored at 4℃[J].LWT-Food Science and Technology,2017,78:317-324.DOI:10.1016/j.lwt.2016.12.035.
[25]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.DOI:10.7506/spkx1002-6630-201824024.
[26]KRAKOVáL,?OLTYS K,BUDI?J,et al.Investigation of bacterial and archaeal communities:novel protocols using modern sequencing by Illumina MiSeq and traditional DGGE-cloning[J].Extremophiles,2016,20(5):795-808.DOI:10.1007//s00792-016-0855-5.
[27]姚鑫,赵爱静,杜苏萍,等.酸性电解水冰对小黄鱼品质及肌肉组织中酶活力变化的影响[J].食品科学,2017,38(13):244-250.DOI:10.7506/spkx1002-6630-201713040.
[28]ERIKSON U,MISIMIL E,JORNE G.Superchilling of rested Atlantic salmon:different chilling strategies and effects on fish and fillet quality[J].Food Chemistry,2011,127(4):1427-1437.DOI:10.1016/j.foodchem.2011.01.036.
[29]杨俊花,赵志辉,郭文博,等.应用Illumina-MiSeq高通量测序技术分析脱氧雪腐镰刀菌烯醇对小鼠肠道菌群的影响[J].动物营养学报,2017,29(1):158-167.DOI:10.3969/j.issn.1006-267x.2017.01.018.
[30]YANG S P,XIE J,QIAN Y F.Determination of spoilage microbiota of pacific white shrimp during ambient and cold storage using nextgeneration sequencing and culture-dependent method[J].Journal of Food Science,2017,82(5):1178-1183.DOI:10.1111/1750-3841.13705.
[31]SALAZAR J K,WANG Y,YU S,et al.Polymerase chain reactionbased serotyping of pathogenic bacteria in food[J].Journal of Microbiological Methods,2015,110:18-26.DOI:10.1016/j.mimet.2015.01.009.
[32]高磊,谢晶,叶藻,等.冷鲜鸡腿肉中优势腐败菌的分离鉴定及腐败能力研究[J].食品与发酵工业,2015,41(8):48-53.DOI:10.13995/j.cnki.11-1802/ts.201508009.
[33]杨秀霞.南极细菌Moritella sp.低温脂肪酶基因的克隆与表达[D].青岛:中国海洋大学,2004:13.
[34]杨娟,郝志成,张亚平,等.基于MiSeq测序分析新疆泥火山土壤细菌群落多样性[J].微生物学通报,2016,43(12):2609-2618.DOI:10.13344/j.microbiol.china.151073.
[2]江艳华,王联珠,许东勤,等.基于高通量测序分析虾夷扇贝柱菌群结构及腐败优势菌[J].食品科学,2018,39(20):140-145.DOI:10.7506/spkx1002-6630-201820021.
[3]MOLINA B,SáEZ M I,MARTíNEZ T F,et al.Effect of ultraviolet light treatment on microbial contamination,some textural and organoleptic parameters of cultured sea bass fillets(Dicentrarchus labrax)[J].Innovative Food Science and Emerging Technologies,2014,26:205-213.DOI:10.1016/j.ifset.2014.07.002.
[4]LIN H M,DENG S G,HUANG S B,et al.Effects of precooling with slurry ice on the quality and microstructure of anglerfish(Lophius americanus)liver[J].Journal of Food Process Engineering,2015,39(1):3-10.DOI:10.1111/jfpe.12193.
[5]ZHANG B,DENG S G,GAO M,et al.Effect of slurry ice on the functional properties of proteins related to quality loss during skipjack tuna(Katsuwonus pelamis)chilled storage[J].Journal of Food Science,2015,80(4):695-702.DOI:10.1111/1750-3841.12812.
[6]XUAN X T,FAN Y F,LING J G,et al.Preservation of squid by slightly acidic electrolyzed water ice[J].Food Control,2017,73:1483-1489.DOI:10.1016/j.foodcont.2016.11.013.
[7]郭全友,董艺伟,李保国,等.淡腌青鱼中木糖葡萄球菌碳源代谢能力研究[J].农业机械学报,2016,47(10):258-266.DOI:10.6041/j,issn.1000-1298.2016.10.032.
[8]薛明,何瑶瑶,邱孟德,等.高通量测序分析凡纳滨对虾育苗期水体菌群结构特征[J].水产学报,2017,41(5):785-794.DOI:10.1196/jfc.20160210291.
[9]WU L Y,WEN C Q,QIN Y J,et al.Phasing amplicon sequencing on Illumina Miseq for robust environmental microbial community analysis[J].BMC Microbiology,2015,15:1-12.DOI:10.1186/s12866-015-0450-4.
[10]DI BELLA B J M,BAO Y G,GLOOR G B,et al.High throughput sequencing methods and analysis for microbiome research[J].Journal of Microbiological Methods,2013,95(3):401-414.DOI:10.1016/j.mimet.2013.08.011.
[11]YANG L Y,LIU S Y,DING J M,et al.Gut microbiota comicroevolution with selection for host humoral immunity[J].Frontiers in Microbiology,2017,8:1-11.DOI:10.3389/fmicb.2017.01243.
[12]XIA X L,ZHANG Q W,ZHANG B,et al.Insights into the biogenic amine metabolic landscape during industrial semidry Chinese rice wine fermentation[J].Journal of Agricultural and Food Chemistry,2016,64(39):7385-7393.DOI:10.1021/acs.jafc.6b01523.
[13]米其利,李雪梅,管莹,等.高通量测序在食品微生物生态学研究中的应用[J].食品科学,2016,37(23):302-308.DOI:10.7506/spkx1002-6630-201623049.
[14]张和平,郑艺.高通量测序技术在乳制品研究中的应用[J].中国食品学报,2015,15(3):1-7.DOI:10.16429/j.1009-7848.2015.03.001.
[15]程三红,汤海青,欧昌荣,等.鲐鱼和大黄鱼冷藏期间体表细菌群落组成和代谢功能的比较分析[J].食品科学,2018,39(19):218-225.DOI:10.7506/spkx1002-6630-201819034.
[16]邓晓影,张宾,汤贺,等.基于高通量测序的南美白对虾中微生物群落分析[J].食品科学,2018,39(24):149-155.DOI:10.7506/spkx1002-6630-201824023.
[17]于美娟,谭欢,马美湖,等.传统固态发酵鱼中细菌群落多样性与品质特征分析[J].食品科学,2017,38(8):86-95.DOI:10.7506/spkx1002-6630-201708015.
[18]农业部.水产品中挥发性盐基氮的测定:SC/T 3032-2007[S].北京:中国标准出版社,2007:3.
[19]杨文鸽,薛长湖,徐大伦,等.大黄鱼冰藏期间ATP关联物含量变化及其鲜度评价[J].农业工程学报,2007,23(6):217-222.
[20]温崇庆,何瑶瑶,薛明,等.高通量测序分析DNA提取引起的对虾肠道菌群结构偏差[J].微生物学报,2016,56(1):130-142.
[21]CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QIIMEallows analysis of high-throughput community sequencing data[J].Nature Methods,2010,7(5):335-336.DOI:10.1038/nmeth.f.303.
[22]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.
[23]CAI L Y,WU X S,LI X X,et al.Effects of different freezing treatments on physicochemical responses and microbial characteristics of Japanese sea bass(Lateolabrax japonicas)fillets during refrigerated storage[J].LWT-Food Science and Technology,2014,15(1):122-129.DOI:10.1016/j.lwt.2014.04.062.
[24]LI Q,ZHANG L T,LU H,et al.Comparison of postmortem changes in ATP-related compounds,protein degradation and endogenous enzyme activity of white muscle and dark muscle from common carp(Cyprinus carpio)stored at 4℃[J].LWT-Food Science and Technology,2017,78:317-324.DOI:10.1016/j.lwt.2016.12.035.
[25]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.DOI:10.7506/spkx1002-6630-201824024.
[26]KRAKOVáL,?OLTYS K,BUDI?J,et al.Investigation of bacterial and archaeal communities:novel protocols using modern sequencing by Illumina MiSeq and traditional DGGE-cloning[J].Extremophiles,2016,20(5):795-808.DOI:10.1007//s00792-016-0855-5.
[27]姚鑫,赵爱静,杜苏萍,等.酸性电解水冰对小黄鱼品质及肌肉组织中酶活力变化的影响[J].食品科学,2017,38(13):244-250.DOI:10.7506/spkx1002-6630-201713040.
[28]ERIKSON U,MISIMIL E,JORNE G.Superchilling of rested Atlantic salmon:different chilling strategies and effects on fish and fillet quality[J].Food Chemistry,2011,127(4):1427-1437.DOI:10.1016/j.foodchem.2011.01.036.
[29]杨俊花,赵志辉,郭文博,等.应用Illumina-MiSeq高通量测序技术分析脱氧雪腐镰刀菌烯醇对小鼠肠道菌群的影响[J].动物营养学报,2017,29(1):158-167.DOI:10.3969/j.issn.1006-267x.2017.01.018.
[30]YANG S P,XIE J,QIAN Y F.Determination of spoilage microbiota of pacific white shrimp during ambient and cold storage using nextgeneration sequencing and culture-dependent method[J].Journal of Food Science,2017,82(5):1178-1183.DOI:10.1111/1750-3841.13705.
[31]SALAZAR J K,WANG Y,YU S,et al.Polymerase chain reactionbased serotyping of pathogenic bacteria in food[J].Journal of Microbiological Methods,2015,110:18-26.DOI:10.1016/j.mimet.2015.01.009.
[32]高磊,谢晶,叶藻,等.冷鲜鸡腿肉中优势腐败菌的分离鉴定及腐败能力研究[J].食品与发酵工业,2015,41(8):48-53.DOI:10.13995/j.cnki.11-1802/ts.201508009.
[33]杨秀霞.南极细菌Moritella sp.低温脂肪酶基因的克隆与表达[D].青岛:中国海洋大学,2004:13.
[34]杨娟,郝志成,张亚平,等.基于MiSeq测序分析新疆泥火山土壤细菌群落多样性[J].微生物学通报,2016,43(12):2609-2618.DOI:10.13344/j.microbiol.china.151073.