电子束辐照对李斯特菌生理特性的影响
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摘要
以英诺克李斯特菌为研究对象,探讨了不同电子束辐照剂量(0.00、0.75、1.50、2.25、3.00、3.75、5.00 kGy)对李斯特菌的灭活作用和生理状态(包括细胞膜通透性、胞内离子泄漏量、Na~+K~+-ATP酶和Ca~(2+)Mg~(2+)-ATP酶活性、膜电位、菌悬液pH值)的影响。结果显示:经5.00 kGy电子束辐照后,李斯特菌菌落总数(N)的对数lg N由对照组的8.9降至5.00 kGy时的1.4,D_(10)=0.69 kGy,杀菌效果显著。随着剂量的增加,细胞膜通透性显著增大、胞内离子(K~+、Ca~(2+)和Mg~(2+))泄漏量显著增加、Na~+K~+-ATP酶和Ca~(2+)Mg~(2+)-ATP酶活性显著降低、膜电位及菌悬液pH值显著下降。以上结果表明,电子束辐照造成李斯特菌生理代谢紊乱,从而加速菌体的死亡。
Listeria innocua was used to investigate the effects of different electron beam irradiation dosages(0.00, 0.75, 1.50, 2.25, 3.00, 3.75 and 5.00 kGy) on the inactivation and physiological status of Listeria innocua(including cell membrane permeability, leakage of intracellular ions, activity of Na~+K~+-ATPase and Ca~(2+)Mg~(2+)-ATPase, membrane potential, pH of bacterial suspension). The results show that the logarithm of total number of Listeria innocua decreases from 8.9 to 1.4 after 5.00 kGy electron beam irradiation with D_(10)=0.69 kGy, and the bactericidal effect is significant. With the increase of dosage, the cell membrane permeability and the leakage of intracellular ions(K~+, Ca~(2+ )and Mg~(2+)) increase significantly. The activities of Na~+K~+-ATPase and Ca~(2+)Mg~(2+)-ATPase decrease significantly, and membrane potential and the pH of bacterial suspension also decrease significantly. According to the above results, electron beam irradiation causes the disorders of physiological metabolism of Listeria innocua, which accelerates the death of the bacteria.
Listeria innocua was used to investigate the effects of different electron beam irradiation dosages(0.00, 0.75, 1.50, 2.25, 3.00, 3.75 and 5.00 kGy) on the inactivation and physiological status of Listeria innocua(including cell membrane permeability, leakage of intracellular ions, activity of Na~+K~+-ATPase and Ca~(2+)Mg~(2+)-ATPase, membrane potential, pH of bacterial suspension). The results show that the logarithm of total number of Listeria innocua decreases from 8.9 to 1.4 after 5.00 kGy electron beam irradiation with D_(10)=0.69 kGy, and the bactericidal effect is significant. With the increase of dosage, the cell membrane permeability and the leakage of intracellular ions(K~+, Ca~(2+ )and Mg~(2+)) increase significantly. The activities of Na~+K~+-ATPase and Ca~(2+)Mg~(2+)-ATPase decrease significantly, and membrane potential and the pH of bacterial suspension also decrease significantly. According to the above results, electron beam irradiation causes the disorders of physiological metabolism of Listeria innocua, which accelerates the death of the bacteria.
引文
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[12]贾倩.电子束和γ射线辐照在素鸡保鲜中的比较研究[D].北京:中国农业科学院,2012.
[13]JEONG S G,KANG D H.Inactivation of escherichia coli O157∶H7,salmonella typhimurium,and Listeria monocytogenes in ready-to-bake cookie dough by gamma and electron beam irradiation[J].Food Microbiology,2017,64:172-178.
[14]NAM K C,KO K Y,MIN B R,et al.Influence of rosemary-tocopherol/packaging combination on meat quality and the survival of pathogens in restructured irradiated pork loins[J].Meat Science,2006,74(2):380-387.
[15]陆海霞,黄小鸣,朱军莉.超高压对单增李斯特菌细胞膜的损伤和致死机理[J].微生物学报,2014,54(7):746-753.LU Haixia,HUANG Xiaoming,ZHU Junli.Effect of ultra high pressure on cell membrane destruction and death of Listeria monocytogenes[J].Acta Microbiologica Sinica,2014,54(7):746-753(in Chinese).
[16]GARCIR-GONZALEZ L,GEERAER A H,MAST J,et al.Membrane permeabilization and cellular death of Escherichia coli,Listeria monocytogenes,and Saccharomyces cerevisiae,as induced by high pressure carbon dioxide treatment[J].Food Microbiology,2010,27(4):541-549.
[17]孔晓雪,付勇,姬赛赛,等.超高压对大肠杆菌O157∶H7细胞膜的损伤效应[J].食品科学,2017,38(8):1-5.KONG Xiaoxue,FU Yong,JI Saisai,et al.Cell membrane injury of Escherichia coli O157∶H7caused by hydrostatic pressure[J].Food Science,2017,38(8):1-5(in Chinese).
[18]FIESTER S E,HELFINSTINE S L,REDFEAM JC,et al.Electron beam irradiation dose dependently damages the bacillus spore coat and spore membrane[J].International Journal of Microbiology,2012,2012(4):579-593.
[19]MILILLO S R,MARTIN E,MUTHAIYAN A,et al.Immediate reduction of Salmonella enterica serotype typhimurium viability via membrane destabilization following exposure to multiplehurdle treatments with heated,acidified organic acid salt solutions[J].Applied and Environmental Microbiology,2011,77(11):3 765-3 772.
[20]吴平,马海乐.脉冲磁场处理对格氏李斯特菌钙离子跨膜行为的影响[J].中国食品学报,2017,17(4):182-188.WU Ping,MA Haile.Effect of pulsed magnetic fields on Ca2+transmembrane behaviors of L.grayi[J].Journal of Chinese Institute of Food Science and Technology,2017,17(4):182-188(in Chinese).
[21]CHEN S,YIN H,TANG S,et al.Metabolic biotransformation of copper-benzo[a]pyrene combined pollutant on the cellular interface of Stenotrophomonas maltophilia[J].Bioresource Technology,2016,204:26-31.
[22]付勇.超高压致大肠杆菌O157∶H7损伤机制的研究[D].南京:南京师范大学,2015.
[23]MZEOVL,VLKOVICˇOVJ,KALOCˇ-AYOVB,et al.Effects ofγ-irradiation on Na,K-ATPase in cardiac sarcolemma[J].Molecular and Cellular Biochemistry,2014,388(1-2):241-247.
[24]MATSUDA H,GINBIN F J,REDDY V N.The effect of X-irradiation on Na-K ATPase and cation distribution in rabbit lens[J].Investigative Ophthalmology and Visual Science,1982,22(2):180-185.
[25]张赟彬,刘笑宇,姜萍萍,等.肉桂醛对大肠杆菌和金黄色葡萄球菌的抑菌作用及抑菌机理研究[J].现代食品科技,2015(5):31-35.ZHANG Yunbin,LIU Xiaoyu,JIANG Pingping,et al.Mechanism and antibacterial activity of cinnamaldehyde against Escherichia coli and Staphylococcus aureus[J].Modern Food Science and Technology,2015(5):31-35(in Chinese).
[26]NOVO D,PERMUTTER N G,HUNT R H,et al.Accurate flow cytometric membrane potential measurement in bacteria using diethyloxacarbocyanine and a ratiometric technique[J].Cytometry,1999,35(1):55-63.
[27]耿飞,王伟,周涛.乌梅提取液对李斯特菌的抑菌机理[J].食品科学,2011,32(15):88-93.GENG Fei,WANG Wei,ZHOU Tao.Antibacterial mechanisms of fructus mume extract against Listeria innocua[J].Food Science,2011,32(15):88-93(in Chinese).
[28]乔支红,程永强,鲁战会,等.乳酸对三种食源性致病菌的抑菌及杀菌作用[J].食品科技,2008(10):187-191.QIAO Zhihong,CHENG Yongqiang,LU Zhanhui,et al.Antibacterial and bactericide activity of lactic acid against three strains of food-borne pathogenic bacteria[J].Food Science and Technology,2008(10):187-191(in Chinese).
[29]刘书成,张良,吉宏武,等.高密度CO2对虾优势腐败菌的杀菌效果及机理[J].农业工程学报,2013,29(14):284-292.LIU Shucheng,ZHANG Liang,JI Hongwu,et al.Sterilizing effect of dense phase carbon dioxide on dominant spoilage bacteria from shrimp and its mechanism[J].Transactions of the Chinese Society of Agricultural Engineering,2013,29(14):284-292(in Chinese).
[30]顾春晖,郑正,赵永富,等.γ-辐照净化水溶液中亚硝酸盐的研究[J].农业环境科学学报,2005,24(6):1 226-1 228.GU Chunhui,ZHENG Zheng,ZHAO Yongfu,et al.Elimination of nitrite in water solution byγ-ray irradiation[J].Journal of Agro-Environment Science,2005,24(6):1 226-1 228(in Chinese).
[31]BORRELY S I,CRUZ A C,MASTRO N L D,et al.Radiation processing of sewage and sludge:A review[J].Progress in Nuclear Energy,1998,33(1-2):3-21.
[32]SHEHATA M M K,GOMAA F A M,HELALZ H.Effects of gamma and electron beam irradiation on viability and DNA elimination of Staphylococcus aureus[J].Archives of Clinical Microbiology,2015,2(6):1-9.
[2]王耀,曹际娟,闫平平,等.食品中3种致病李氏菌MPCR-DHPLC检测方法的建立[J].食品科学,2010,31(8):158-162.WANG Yao,CAO Jijuan,YAN Pingping,et al.MPCR-DHPLC detection of three species of pathogenic Listeria in food[J].Food Science,2010,31(8):158-162(in Chinese).
[3]CHEN J,ZHANG X,MEI L,et al.Prevalence of Listeriain Chinese food products from 13provinces between 2000and 2007and virulence characterization of Listeria monocytogenes isolates[J].Foodborne Pathogens and Disease,2009,6(1):7-14.
[4]齐颖颖,吴萌,王怡雯,等.单增李斯特菌单克隆抗体的研制及特性分析[J].食品与生物技术学报,2016,35(2):151-155.QI Yingying,WU Meng,WANG Yiwen,et al.Preparation and characterization of monoclonal antibodies against Listeria monocytogene[J].Journal of Food Science and Biotechnology,2016,35(2):151-155(in Chinese).
[5]靳晓燕,韩军,于宏伟,等.食品中单核增生性李斯特菌(Listeria monocytogenes)污染状况研究[J].中国食品学报,2009,9(1):226-231.JIN Xiaoyan,HAN Jun,YU Hongwei,et al.The survey on the contamination situation of Listeria monocytogenes in foods[J].Journal of Chinese Institute of Food Science and Technology,2009,9(1):226-231(in Chinese).
[6]汪勋清,哈益明,高美须.食品辐照加工技术[M].北京:化学工业出版社,2005.
[7]施培新.食品辐照加工原理与技术[M].北京:中国农业科学技术出版社,2004.
[8]HARDER M N C,ARTHUR V,ARTHUR PB.Irradiation of foods:Processing technology and effects on nutrients:Effect of ionizing radiation on food components[J].Encyclopedia of Food and Health,2016.doi:1016/B978-0-12-384947-2.00405-0.
[9]LACROIX M,FOLLETT P.Combination irradiation treatments for food safety and phytosanitary uses[J].Stewart Postharvest Review,2015,11(3):1-10.
[10]UNLUTURK S.Impact of irradiation on the microbial ecology of foods[M]∥Quantitative Microbiology in Food Processing.[S.l.]:[s.n.],2016.
[11]SONG H P,KIM B,JUNG S,et al.Effect of gamma and electron beam irradiation on the survival of pathogens inoculated into salted,seasoned,and fermented oyster[J].LWT-Food Science and Technology,2009,42(8):1 320-1 324.
[12]贾倩.电子束和γ射线辐照在素鸡保鲜中的比较研究[D].北京:中国农业科学院,2012.
[13]JEONG S G,KANG D H.Inactivation of escherichia coli O157∶H7,salmonella typhimurium,and Listeria monocytogenes in ready-to-bake cookie dough by gamma and electron beam irradiation[J].Food Microbiology,2017,64:172-178.
[14]NAM K C,KO K Y,MIN B R,et al.Influence of rosemary-tocopherol/packaging combination on meat quality and the survival of pathogens in restructured irradiated pork loins[J].Meat Science,2006,74(2):380-387.
[15]陆海霞,黄小鸣,朱军莉.超高压对单增李斯特菌细胞膜的损伤和致死机理[J].微生物学报,2014,54(7):746-753.LU Haixia,HUANG Xiaoming,ZHU Junli.Effect of ultra high pressure on cell membrane destruction and death of Listeria monocytogenes[J].Acta Microbiologica Sinica,2014,54(7):746-753(in Chinese).
[16]GARCIR-GONZALEZ L,GEERAER A H,MAST J,et al.Membrane permeabilization and cellular death of Escherichia coli,Listeria monocytogenes,and Saccharomyces cerevisiae,as induced by high pressure carbon dioxide treatment[J].Food Microbiology,2010,27(4):541-549.
[17]孔晓雪,付勇,姬赛赛,等.超高压对大肠杆菌O157∶H7细胞膜的损伤效应[J].食品科学,2017,38(8):1-5.KONG Xiaoxue,FU Yong,JI Saisai,et al.Cell membrane injury of Escherichia coli O157∶H7caused by hydrostatic pressure[J].Food Science,2017,38(8):1-5(in Chinese).
[18]FIESTER S E,HELFINSTINE S L,REDFEAM JC,et al.Electron beam irradiation dose dependently damages the bacillus spore coat and spore membrane[J].International Journal of Microbiology,2012,2012(4):579-593.
[19]MILILLO S R,MARTIN E,MUTHAIYAN A,et al.Immediate reduction of Salmonella enterica serotype typhimurium viability via membrane destabilization following exposure to multiplehurdle treatments with heated,acidified organic acid salt solutions[J].Applied and Environmental Microbiology,2011,77(11):3 765-3 772.
[20]吴平,马海乐.脉冲磁场处理对格氏李斯特菌钙离子跨膜行为的影响[J].中国食品学报,2017,17(4):182-188.WU Ping,MA Haile.Effect of pulsed magnetic fields on Ca2+transmembrane behaviors of L.grayi[J].Journal of Chinese Institute of Food Science and Technology,2017,17(4):182-188(in Chinese).
[21]CHEN S,YIN H,TANG S,et al.Metabolic biotransformation of copper-benzo[a]pyrene combined pollutant on the cellular interface of Stenotrophomonas maltophilia[J].Bioresource Technology,2016,204:26-31.
[22]付勇.超高压致大肠杆菌O157∶H7损伤机制的研究[D].南京:南京师范大学,2015.
[23]MZEOVL,VLKOVICˇOVJ,KALOCˇ-AYOVB,et al.Effects ofγ-irradiation on Na,K-ATPase in cardiac sarcolemma[J].Molecular and Cellular Biochemistry,2014,388(1-2):241-247.
[24]MATSUDA H,GINBIN F J,REDDY V N.The effect of X-irradiation on Na-K ATPase and cation distribution in rabbit lens[J].Investigative Ophthalmology and Visual Science,1982,22(2):180-185.
[25]张赟彬,刘笑宇,姜萍萍,等.肉桂醛对大肠杆菌和金黄色葡萄球菌的抑菌作用及抑菌机理研究[J].现代食品科技,2015(5):31-35.ZHANG Yunbin,LIU Xiaoyu,JIANG Pingping,et al.Mechanism and antibacterial activity of cinnamaldehyde against Escherichia coli and Staphylococcus aureus[J].Modern Food Science and Technology,2015(5):31-35(in Chinese).
[26]NOVO D,PERMUTTER N G,HUNT R H,et al.Accurate flow cytometric membrane potential measurement in bacteria using diethyloxacarbocyanine and a ratiometric technique[J].Cytometry,1999,35(1):55-63.
[27]耿飞,王伟,周涛.乌梅提取液对李斯特菌的抑菌机理[J].食品科学,2011,32(15):88-93.GENG Fei,WANG Wei,ZHOU Tao.Antibacterial mechanisms of fructus mume extract against Listeria innocua[J].Food Science,2011,32(15):88-93(in Chinese).
[28]乔支红,程永强,鲁战会,等.乳酸对三种食源性致病菌的抑菌及杀菌作用[J].食品科技,2008(10):187-191.QIAO Zhihong,CHENG Yongqiang,LU Zhanhui,et al.Antibacterial and bactericide activity of lactic acid against three strains of food-borne pathogenic bacteria[J].Food Science and Technology,2008(10):187-191(in Chinese).
[29]刘书成,张良,吉宏武,等.高密度CO2对虾优势腐败菌的杀菌效果及机理[J].农业工程学报,2013,29(14):284-292.LIU Shucheng,ZHANG Liang,JI Hongwu,et al.Sterilizing effect of dense phase carbon dioxide on dominant spoilage bacteria from shrimp and its mechanism[J].Transactions of the Chinese Society of Agricultural Engineering,2013,29(14):284-292(in Chinese).
[30]顾春晖,郑正,赵永富,等.γ-辐照净化水溶液中亚硝酸盐的研究[J].农业环境科学学报,2005,24(6):1 226-1 228.GU Chunhui,ZHENG Zheng,ZHAO Yongfu,et al.Elimination of nitrite in water solution byγ-ray irradiation[J].Journal of Agro-Environment Science,2005,24(6):1 226-1 228(in Chinese).
[31]BORRELY S I,CRUZ A C,MASTRO N L D,et al.Radiation processing of sewage and sludge:A review[J].Progress in Nuclear Energy,1998,33(1-2):3-21.
[32]SHEHATA M M K,GOMAA F A M,HELALZ H.Effects of gamma and electron beam irradiation on viability and DNA elimination of Staphylococcus aureus[J].Archives of Clinical Microbiology,2015,2(6):1-9.