天然植物精油的抑菌活性及其作用机理研究
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摘要
本论文研究了植物单精油以及复配精油对四种食品微生物(大肠杆菌,金黄色葡萄球菌,枯草芽孢杆菌和酿酒酵母)的抑制活性。使用滤纸片法和试管梯度稀释法对十种植物精油的抑菌活性进行了初步筛选。结果显示,牛至精油对四种供试菌种均显示出了最强的抑制活性,罗勒精油和佛手柑精油对革兰氏阳性菌有很好的抑制效果,而紫苏精油则对酿酒酵母有很强的抑制专一性。采用棋盘稀释法,研究复配精油间可能存在的协同增效作用。以FIC指数作为参照依据,选定的复配精油组合及联合最小抑菌浓度如下：大肠杆菌(牛至精油-罗勒精油,MIC联合：0.313-0.313μL/mL)；金黄色葡萄球菌(罗勒精油-佛手柑精油,MIC联合：0.313-0.156μL/mL)；枯草芽孢杆菌(牛至精油-佛手柑精油,MIC联合：0.313-0.313μL/mL)；酿酒酵母(牛至精油-紫苏精油,MIC联合：0.313-0.156μL/mL)。
采用气质联用技术(GC/MS),对所选植物精油的化学组成进行了分析鉴定：牛至精油和罗勒精油的主要化学成分是酚类和萜烯类物质,佛手柑精油中主要含有萜烯类、醇类和酯类物质,而酮类物质在紫苏精油中的含量相对较高。
通过观察菌体的扫描电镜图片,测定菌体细胞培养液的电导率、细胞溶出物以及菌体细胞膜脂肪酸的变化,研究了复配精油的抑菌机理。结果表明,所有的复配精油均能够破坏菌体细胞膜的完整性,并使其通透性增加,最终达到抑制菌体生长的目的。
最后,为了改善精油的溶解度以提高抑菌活性,本研究比较了九种增溶剂对精油的增溶效果,并对增溶体系的抑菌活性和抑菌机理进行了研究。结果发现,HP-β-CD对精油的增溶效果最为明显。随着HP-β-CD的加入,可以显著降低复配精油的用量,提高了复配精油对菌体的作用强度。
The aim of this study was to evaluate the antimicrobial efficacy of selected essential oil (EO) and their combinations against four food-related microorganisms. Ten EOs were initially screened against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Saccharomyces cerevisiae using agar disc diffusion and broth dilution methods. The highest efficacy against all the tested strains was shown when testing the oregano EO. EOs of basil and bergamot were active against the Gram-positive bacteria, while perilla EO strongly inhibited the growth of S.cerevisiae. The checkerboard method was then used to investigate the antimicrobial efficacy of EO combinations by means of the fractional inhibitory concentration index (FICI). Based on an overall consideration of antimicrobial activity, organoleptic impact and cost, four EO combinations were selected and their MIC values were listed as follows:oregano-basil (0.313-0.313 [μL/mL) for E. coli, basil-bergamot (0.313-0.156μL/mL) for S. aureus, oregano-bergamot (0.313-0.313μL/mL) for B. subtilis and oregano-perilla (0.313-0.156μL/mL) for S. cerevisiae.
The chemical components of selected EOs were then analyzed by GC/MS. Phenols and terpenes were the major antimicrobial compounds in oregano and basil EOs. The dominant active components of bergamot EO were alcohols, esters and terpenes. For perilla EO, the major active constituents were mainly ketones.
Furthermore, the mechanisms of the antimicrobial action of EO combinations were studied by the measurement of conductivity and the release of cell constituents, the electronic microscopy observations of the cells and the changes in the percentage of principal fatty acids of cells. The results demonstrated that all EO combinations destroyed the integrity of cell membrane, resulting in the death of microorganisms.
Finally, we test the ability of nine solubilizers to increase EOs solubility and the effect of selected solubilizer on the antimicrobial activity of EO combinations by checkerboard method. The results indicated that the effect of HP-β-CD on increasing EOs solubility was most obvious. The addition of HP-β-CD could not only decrease the usage of EO combinations significantly, but also enhance the effects of EO combinations on microorganisms.
采用气质联用技术(GC/MS),对所选植物精油的化学组成进行了分析鉴定：牛至精油和罗勒精油的主要化学成分是酚类和萜烯类物质,佛手柑精油中主要含有萜烯类、醇类和酯类物质,而酮类物质在紫苏精油中的含量相对较高。
通过观察菌体的扫描电镜图片,测定菌体细胞培养液的电导率、细胞溶出物以及菌体细胞膜脂肪酸的变化,研究了复配精油的抑菌机理。结果表明,所有的复配精油均能够破坏菌体细胞膜的完整性,并使其通透性增加,最终达到抑制菌体生长的目的。
最后,为了改善精油的溶解度以提高抑菌活性,本研究比较了九种增溶剂对精油的增溶效果,并对增溶体系的抑菌活性和抑菌机理进行了研究。结果发现,HP-β-CD对精油的增溶效果最为明显。随着HP-β-CD的加入,可以显著降低复配精油的用量,提高了复配精油对菌体的作用强度。
The aim of this study was to evaluate the antimicrobial efficacy of selected essential oil (EO) and their combinations against four food-related microorganisms. Ten EOs were initially screened against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Saccharomyces cerevisiae using agar disc diffusion and broth dilution methods. The highest efficacy against all the tested strains was shown when testing the oregano EO. EOs of basil and bergamot were active against the Gram-positive bacteria, while perilla EO strongly inhibited the growth of S.cerevisiae. The checkerboard method was then used to investigate the antimicrobial efficacy of EO combinations by means of the fractional inhibitory concentration index (FICI). Based on an overall consideration of antimicrobial activity, organoleptic impact and cost, four EO combinations were selected and their MIC values were listed as follows:oregano-basil (0.313-0.313 [μL/mL) for E. coli, basil-bergamot (0.313-0.156μL/mL) for S. aureus, oregano-bergamot (0.313-0.313μL/mL) for B. subtilis and oregano-perilla (0.313-0.156μL/mL) for S. cerevisiae.
The chemical components of selected EOs were then analyzed by GC/MS. Phenols and terpenes were the major antimicrobial compounds in oregano and basil EOs. The dominant active components of bergamot EO were alcohols, esters and terpenes. For perilla EO, the major active constituents were mainly ketones.
Furthermore, the mechanisms of the antimicrobial action of EO combinations were studied by the measurement of conductivity and the release of cell constituents, the electronic microscopy observations of the cells and the changes in the percentage of principal fatty acids of cells. The results demonstrated that all EO combinations destroyed the integrity of cell membrane, resulting in the death of microorganisms.
Finally, we test the ability of nine solubilizers to increase EOs solubility and the effect of selected solubilizer on the antimicrobial activity of EO combinations by checkerboard method. The results indicated that the effect of HP-β-CD on increasing EOs solubility was most obvious. The addition of HP-β-CD could not only decrease the usage of EO combinations significantly, but also enhance the effects of EO combinations on microorganisms.
引文
[1]杜丽娜,张存莉,朱玮,等.植物次生代谢合成途径及生物学意义[J].西北林学院学报,2005,20(3)：150-155
[2]刘洪波,史东辉,张瑜.植物精油抗菌活性研究进展[J].畜牧与饲料科学,2009,30(1)：12-14
[3]高献礼,李超.植物性香料提取技术的研究进展[J].中国食品添加剂,2006,6:134-138
[4]焦启源.芳香植物极其利用(上册)[M].上海：上海科学技术出版社,1963
[5]金琦.香料生产工艺学[M].哈尔滨：东北林业大学出版,1996
[6]张坚,孙培龙,杨开,等.植物性天然香料的提取与应用[J].食品研究与开发,2006,27(4):181-184
[7]郭秀艳.臭柏、油松精油的提取与抑菌活性[D].内蒙古：内蒙古农业大学博士学位论文,2009
[8]孙鹏,刘文文.植物精油提取和应用的研究进展[J].甘肃科技,2007,23(5):139-141
[9]毕艳红,乔旭光,田云.分子蒸馏技术分离纯化大蒜精油的研究[J].分离与提取,2007,33(5)：153-157
[10]周峰.香辛料精油中抗菌成分分离鉴定、功能协同强化及量化研究[D].北京：中国农业大学博士学位论文,2007
[11]董丽,邢钧,吴采樱.香精香料的分析方法进展[J].分析科学学报,2003,19(2):188-192
[12]王颖,姜子涛,李荣,等.黑胡椒和白胡椒精油抗氧化性能及清除自由基能力的比较[J].中国调味品,2009,34(5):50-54
[13]冯雪,姜子涛,李荣,等.天然调味香料众香子精油的抗氧化性能及清除自由基能力研究[J].中国调味品,2010,35(4):52-55
[14]顾仁勇,张石峰,刘莹莹,等.五种香辛料精油抑菌及抗氧化性能研究[J].食品科学,2008,29(3)：106-108
[15]李荣,姜子涛,马丽.月桂精油抗氧化性能及清除自由基能力的研究[J].中国调味品,2009,34(11)：58-63
[16]丁慧.芳香疗法与中医药理论渊源初探[J].中华中医药杂志,2006,21(7):434
[17]黄汉超,陈宏硅,吴伟.中医药治疗不稳定型心绞痛的临床研究与治疗进展[J].中医药学刊,2006,24(1)：101-104
[18]林木振,王孟和,刘秉忠.牡丹皮散联合阿司匹林治疗腰源性腹痛56例[J].浙江中西医结合杂志,2009,19(1)：10-11
[19]高健,段大航,蔡秀荣.大蒜油对心血管作用研究现状[J].社区医学杂志,2007,5(15)：53-55
[20]刘红杰,金若敏.薄荷油研究进展[J].山东中医药大学学报,2006,30(6):502-505
[21]杨帆,王羽梅.山鸡椒(山苍子)精油研究现状[J].韶关学院学报(自然科学),2009,30(6):80-84
[22]赵丽琴.山胡椒属萜类及其生物活性研究进展[J].时珍国医国药,2006,17(2):171-174
[23]潘年松,盛勇,张学愈.莪术抗肿瘤作用与品种和加工工艺的关系[J].中国中医药信息杂志,2006,13(8)：35-36
[24]Keita S M, Vincent C, Schmit J, et al. Efficacy of essential oil of Ocimum basilicum L. and O. gratissimum L. applied as an insecticidal fumigant and powder to control Callosobruchus maculates [J]. Journal of Stored Products Research,2001,37:339-349
[25]Kim S I, Park C, Ohh M H, et al. Contact and fumigant activities of aromatic plant extracts and essential oils against Lasioderma serricorne [J]. Journal of Stored Products Research,2003,39:11-19
[26]Aslan I, Ozbek H, Calmasur O. Toxicity of essential oil vapours to two greenhouse pests, Tetranychus urticae Koch and Bemisia tabaci Genn [J]. Industrial Crops and Products, 2004,19:167-173
[27]艾应伟,范志金,李伟,等.植物源生理活性物质与害虫防治[J].资源科学,2000,22(3)：78-81
[28]钱骅,赵伯涛,夏劲,等.九种精油的抗菌活性及其防腐特性研究[J].中国调味品,2010,35(4):69-72
[29]Valero M, Salmeron M C. Antibacterial activity of 11 essential oils against Bacillus cereus in tyndallized carrot broth [J]. International Journal of Food Microbiology,2003,85: 73-81
[30]何玮,李光武,傅佳.薰衣草及橙花精油体外抑菌作用的研究[J].安徽医药,2010,14(5)：525-527
[31]吴慧清,吴清平,石立三,等.植物精油对微生物的抑菌效果评估研究[J].食品科学,2008,29(12)：83-86
[32]马英姿,谭琴,李恒熠.樟树叶及天竺桂叶的精油抑菌活性研究[J].中南林业科技大学学报,2009,29(1)：36-40
[33]Serge A, David M. Antimicrobial properties of allicin from garlic [J]. Microbes and Infection,1999,1:125-129
[34]钟瑞敏,肖仔君,张振明,等.小茴香籽精油成分及其抗菌活性研究[J].林产化学与工业,2007,27(6)：36-40
[35]陈丽艳,崔志恒.植物精油抗菌活性的研究进展[J].黑龙江医药,2006,19(3):197-198
[36]Hazzit M, Baaliouamer A, Faleiro M L, et al. Composition of the essential oils of Thymus and Origanum species from Algeria and their antioxidant and antimicrobial activities [J]. Journal of Agricultural and Food Chemistry,2006,54:6314-6321
[37]胡艳芬,江文斌,蒋琳琳,等.百里香酚和香芹酚体外抑菌作用的研究[J].兽药与饲料,2010,10:107
[38]楼兴隆,李晓明,张鞍灵,等.丁香油抑菌成分研究[J].西北农业学报,2006,15(3):91-93
[39]吕世明,陈杖榴,陈建新,等.丁香酚体外抑菌作用研究[J].食品科学,2008,29(9):122-124
[40]郑国华,刘焱文,陈树和,等.牛至各生长期抗菌成分的分析[J].中国中药杂志,1999,24(9):536-538
[41]张静,冯岗,袁旭超,等.百里香酚抑茵活性初探[J].中国农业学报,2009,25(21)：277-280
[42]王雪梅,谌徽,李雪姣,等.天然活性单萜—柠檬烯的抑菌性能研究[J].吉林农业大学学报,2010,32(1)：24-28
[43]李京晶,籍保平,周峰,等.丁香和肉桂挥发油的提取、主要成分测定及其抗菌活性研究[J].食品科学,2006,27(8):64-68
[44]钱昆,周涛.植物源天然防腐剂应用和机理研究的最新进展[J].中国食品添加剂,2006,5：100-104
[45]NCCLS. Methods for Dilution Antimicrobial Susceptibility Tests for Bacterict That Grow Aerobically: Approved Standard-Sixth Edition [M]. National Committee for Clinical Laboratory Standards,2002
[46]Reza S, Rahman A, Lee J, et al. Potential roles of essential oil and organic extracts of Zizyphus jujube in inhibiting food-borne pathogens [J]. Food Chemistry,2010,119: 981-986
[47]Davidson P M, Parish M E.. Methods for testing the efficacy of food antimicrobials [J]. Food Technology,1989,43:148-155
[48]李宏.精油的抗病毒和抗微生物作用[J].香料香精化妆品,2002,5:5-8
[49]王帆,杨静东,王春梅,等.复配植物源杀菌剂的开发研究[J].江西农业学报,2010,22(2):87-89
[50]顾仁勇,傅伟昌,银永忠.丁香和肉桂精油联合抗菌作用初步研究[J].食品科学,2008,29(10)：115-119
[51]吕飞.生物抗菌包装体系及其对黑鱼品质影响的研究[D].浙江：浙江大学博士学位论文,2009
[52]Yamazaki K, Yamamoto T, Kawai Y, et al. Enhancement of antilisterial activity of essential oil constituents by nisin and diglycerol fatty acid ester [J]. Food microbiology, 2004,21:283-289
[53]Turgis M, Han J, Caillet S, et al.. Antimicrobial activity of mustard essential oil against Escherichia coli O157:H7 and Salmonella typhi [J]. Food control,2009,20:1073-1079
[54]Oussalah M, Caillet S, Saucier L, et al. Antimicrobial effects of selected plant essential oils on the growth of a Pseudomonas putida strain isolated from meat [J]. Meat Science, 2006,73:236-244
[55]孙峋汪,汪靖超,李洪涛,等.迷迭香酸的抗菌机理研究[J].青岛大学学报(自然科学版),2005,18(4)：41-45
[56]Pasqua R D, Betts G, Hoskins N, et al. Membrane toxicity of antimicrobial compounds from essential oils [J]. Journal of Agricultural and Food Chemistry,2007,55:4863-4870
[57]郑罡,曹煜,魏羽佳.茶树油对白假丝酵母菌抑菌效果[J].中国消毒学杂志,2007,24(5):438440
[58]顾仁勇.五种香辛料精油的提取及用于冷却猪肉保鲜研究[D].湖南：湖南农业大学硕士学位论文,2007
[59]李鹏霞,张兴,刘亚敏,等.36种精油对采后番茄防腐保鲜活性的影响[J].西北林学院学报,2008,23(3)：156-159
[60]刘芳,张小青,吴三林,等.峨眉含笑精油对冬枣保鲜的研究[J].长春师范学院学报(自然科学版),2008,27(3)：65-69
[61]张春红,刘静,陈丽,等.三种植物精油对霉菌的抑制效果测定及其在大米储藏中应用研究[J].食品科技,2007,32(12):105-108
[62]Shelef L A, Jyothi E K, Bulgarelli M A. Growth of enteropathogenic and spoilage bacteria in sage-containing broth and foods [J]. Journal of Food Science,1984,49:737-740
[63]疏秀林,施庆珊,欧阳友生,等.植物精油的抗菌特性及在食品工业中应用研究新进展[J].生物技术,2006,16(6):89-92
[64]Stammati A, Bonsi P, Zucco F, et al. Toxicity of selected plant volatiles in microbial and mammalian short-term assays [J]. Food and chemical Toxiology,1999(37):813-823
[65]Celiktas O, Kocabas E, Bedir E, et al. Antimicrobial activities of methanol extracts and essential oils of Rosmarinus oficinalis, depending on location and seasonal variations [J]. Food Chemistry,2007,100:553-559
[66]Jacob C, Mathiasen L, Powell D. Designing effective messages for microbial food safety hazards [J]. Food Control,2010,21:1-6
[67]Fleming-Jones M, Smith R. Volatile organic compounds in foods: a five year study [J]. Journal of Agricultural and Food Chemistry,2003,51:8120-8127
[68]Goni P, Lopez P, Sanchez C, et al. Antimicrobial activity in the vapour phase of a combination of cinnamon and clove essential oils [J]. Food Chemistry,2009,116:982-989
[69]Hsieh P C, Mau J L, Huang S H. Antimicrobial effect of various combinations of plant extracts [J]. Food Microbiology,2001,18:35-43
[70]Nazer A I, Kobilinsky A, Tholozan J L, et al. Combinations of food antimicrobials at low levels to inhibit the growth of Salmonella sv. Typhimurium:A synergistic effect? [J]. Food Microbiology,2005,22:391-398
[71]Gutierrez J, Barry-Ryan C, Bourke P. Antimicrobial activity of plant essential oils using food model media: Efficacy, synergistic potential and interactions with food components [J]. Food Microbiology,2009,26:142-150
[72]Firuzi O, Asadollahi M, Gholami M, et al. Composition and biological activities of essential oils from four Heracleum species [J]. Food Chemistry,2010,122:117-122
[73]Rota M C, Carraminana J J, Burillo J, et al. In vitro antimicrobial activity of essential oils from aromatic plants against selected foodborne pathogens [J]. Journal of Food Protection, 2004,67:1252-1256
[74]Weerakkody N S, Caffin N, Turner M S, et al. In vitro antimicrobial activity of less-utilized spice and herb extracts against selected food-borne bacteria [J]. Food Control, 2010,21:1408-1414
[75]Hemaiswarya S, Kruthiventi A K, Doble M. Synergism between natural products and antibiotics against infectious diseases [J]. Phytomedicine,2008,15:639-652
[76]Rota M C, Herrera A, Martinez R M, et al. Antimicrobial activity and chemical composition of Thymus vulgaris, Thymus zygis and Thymus hyemalis essential oils [J]. Food Control,2008,19:681-687
[77]Oussalah M, Caillet S, Saucier L, et al. Antimicrobial effects of selected plant essential oils on the growth of a Pseudomonas putida strain isolated from meat [J]. Meat Science, 2006,73:236-244
[78]Vaara, M. Agents that increase the permeability of the outer membrane [J]. Microbiological Reviews,1992,56:395-411
[79]Elgayyar M, Draughon F A, Golden D A, et al. Antimicrobial activity of essential oils from plants against selected pathogenic and saprophytic microorganisms [J]. Journal of Food Protection,2001,64:1019-1024
[80]Burt S, Reinders R D. Antibacterial activity of selected plant essential oils against Escherichia coli O157.H7 [J]. Applied Microbiology,2003,36:162-167
[81]Gutierrez J, Rodriguez G, Barry-Ryan C, et al. Efficacy of plant essential oils against food-borne pathogens and spoilage bacteria associated with ready to eat vegetables: antimicrobial and sensory screening [J]. Journal of Food Protection,2008,71:1846-1854
[82]Hammer K A, Carson C F, Riley T V. Antimicrobial activity of essential oils and other plant extracts [J]. Journal of Applied Microbiology,1999,86:985-990
[83]Runyoro D, Ngassapa O, Vagionas K, et al. Chemical composition and antimicrobial activity of the essential oils of four Ocimum species growing in Tanzania [J]. Food Chemistry,2010,119:311-316
[84]Grayer R J, Kite G C, Goldstone F J, et al. Infraspecific taxonomy and essential oil chemotypes in sweet basil, Ocimum basilicum[J]. Phytochemistry,1996,43:1033-1039
[85]Friedman M, Henika P R, Levin C E, et al. Antibacterial activities of plant essential oils and their components against escherichia coli O157.H7 and salmonella enterica in Apple Juice [J]. Journal of Agricultural and Food Chemistry,2004,52:6042-6048
[86]Yamamoto H, Ogawa T. Antimicrobial activity of perilla seed polyphenols against oral pathogenic bacteria [J]. Bioscience, Biotechnology, and Biochemistry,2002,66:921-924
[87]Dorman H, Deans S G. Antimicrobial agents from plants: antibacterial activity of plant volatile oils [J]. Journal of Applied Microbiology,2000,88:308-316
[88]Gill A O, Delaquis P, Russo P, et al. Evaluation of antilisterial action of cilantro oil on vacuum packed ham [J]. International Journal of Food Microbiology,2002,73:83-92
[89]Burt S. Essential oils: Their antibacterial properties and potential applications in foods-A review [J]. International Journal of Food Microbiology,2004,94:223-253
[90]Ultee A, Kets E, Alberda M, et al. Adaptation of the food-borne pathogen Bacillus cereus to carvacrol [J]. Archives of Microbiology,2000,174:233-238
[91]Naigre R, Kalck P, Roques C, et al. Comparison of antimicrobial properties of monoterpenes and their carbonylated products [J]. Planta Medica,1996,62:275-277
[92]Uhart M, Maks N, Ravishankar S. Effect of spices on growth and survival of Salmonella typhimurium DT 104 in ground beef stored at 4 and 8℃[J]. Journal of Food Safety,2006, 26:115-125
[93]Devlieghere F, Vermeulen A, Debevere J. Chitosan: antimicrobial activity, interactions with food components and applicability as a coating on fruit and vegetables [J]. Food Microbiology,2004,21:703-714
[94]Adams R P. Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy [M]. Allured Publishing Corporation:USA,2001
[95]Panizzi L, Flamini G, Cioni P L, et al. Composition and antimicrobial properties of essential oils of four Mediterranean Lamiaceae [J]. Journal of Ethnopharmacology,1993, 39:167-170
[96]Ceylan E, Fung D Y C. Antimicrobial activity of spices [J]. Journal of Rapid Methods and Automation in Microbiology,2004,12:1-55
[97]Bozen B, Dukic N M, Simin N, et al. Characterization of the volatile composition of essential oils of some Lamiaceae spices and the antimicrobial and antioxidant activities of the entire oils [J]. Journal of Agricultural and Food Chemistry,2006,54:1822-1828
[98]Vina A, Murillo E. Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia [J]. Journal of the Brazilian Chemical Society,2003,14:744-749
[99]Fang T, Goto M, Sasaki M, et al. Combination of supercritical CO2 and vacuum distillation for the fractionation of bergamot oil [J]. Journal of Agricultural and Food Chemistry,2004,52:5162-5167
[100]Figoli A, Donato L, Carnevale R, et al. Bergamot essential oil extraction by pervaporation [J]. Desalination,2006,193:160-165
[101]Nitta M, Kobayashi H, Ohnishi-Kameyama M, et al. Essential oil variation of cultivated and wild perilla analyzed by GC/MS [J]. Biochemical Systematics and Ecology,2006,34: 25-37
[102]Calvey E M, White K D, Matusil J E, et al. Allium chemistry: identification of organosulfur compounds in ramp (Allium tricoccum) homogenates [J]. Phytochemistry, 1998,49:359-364
[103]吴玉荷,杨小华,蒋灵芝.薇甘菊水提物的抑菌试验研究[J].中国野生植物资源,2007,26(1)：51-51
[104]谢家仪,董光军,刘振英.扫描电镜的微生物样品制备方法[J].电子显微学报,2005,24(4):440
[105]于庆华,殷文政,艾启俊,等.鹿蹄草素对金黄色葡萄球菌和绿脓杆菌的抑制效果观察[J].食品科学,2008,29(4):90-93
[106]杜春明,焦振泉,郭云昌,等.唐菖蒲伯克霍尔德菌菌体脂肪酸成分测定与分析[J].卫生研究,2005,34(5):613-616
[107]王关林,蒋丹,方宏筠.天南星的抑菌作用及其机理研究[J].畜牧兽医学报,2004,35(30)：280-285
[108]Tajkarimi M M, Ibrahima S A, Cliver D O. Antimicrobial herb and spice compounds in food [J]. Food Control,2010,21:1199-1218
[109]Sikkema J, De Bont J A M, Poolman B. Mechanisms of membrane toxicity of hydrocarbons [J]. Microbiological Reviews,1995,59:201-222
[110]Lambert R J W, Skandamis P N, Coote P, et al. A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol [J]. Journal of Applied Microbiology,2001,91:453-462
[111]Carson C F, Mee B J, Riley T V. Mechanism of action of Melaleuca alternifolia (tea tree) oil on Staphylococcus aureus determined by time-kill, lysis, leakage and salt tolerance assays and electron microscopy [J]. Antimicrobial Agents and Chemotherapy,2002,46: 1914-1920
[112]Moreira M R, Ponce A G, Del Valle C E, et al. Inhibitory parameters of essential oils to reduce a foodborne pathogen [J]. LWT-Food Science and Technology,2005,38:565-570
[113]Santiesteban-Lopez A, Palou E, Lopez-Malo A. Susceptibility of food-borne bacteria to binary combinations of antimicrobials at selected aw and pH [J]. Journal of Applied Microbiology,2007,102:486-497
[2]刘洪波,史东辉,张瑜.植物精油抗菌活性研究进展[J].畜牧与饲料科学,2009,30(1)：12-14
[3]高献礼,李超.植物性香料提取技术的研究进展[J].中国食品添加剂,2006,6:134-138
[4]焦启源.芳香植物极其利用(上册)[M].上海：上海科学技术出版社,1963
[5]金琦.香料生产工艺学[M].哈尔滨：东北林业大学出版,1996
[6]张坚,孙培龙,杨开,等.植物性天然香料的提取与应用[J].食品研究与开发,2006,27(4):181-184
[7]郭秀艳.臭柏、油松精油的提取与抑菌活性[D].内蒙古：内蒙古农业大学博士学位论文,2009
[8]孙鹏,刘文文.植物精油提取和应用的研究进展[J].甘肃科技,2007,23(5):139-141
[9]毕艳红,乔旭光,田云.分子蒸馏技术分离纯化大蒜精油的研究[J].分离与提取,2007,33(5)：153-157
[10]周峰.香辛料精油中抗菌成分分离鉴定、功能协同强化及量化研究[D].北京：中国农业大学博士学位论文,2007
[11]董丽,邢钧,吴采樱.香精香料的分析方法进展[J].分析科学学报,2003,19(2):188-192
[12]王颖,姜子涛,李荣,等.黑胡椒和白胡椒精油抗氧化性能及清除自由基能力的比较[J].中国调味品,2009,34(5):50-54
[13]冯雪,姜子涛,李荣,等.天然调味香料众香子精油的抗氧化性能及清除自由基能力研究[J].中国调味品,2010,35(4):52-55
[14]顾仁勇,张石峰,刘莹莹,等.五种香辛料精油抑菌及抗氧化性能研究[J].食品科学,2008,29(3)：106-108
[15]李荣,姜子涛,马丽.月桂精油抗氧化性能及清除自由基能力的研究[J].中国调味品,2009,34(11)：58-63
[16]丁慧.芳香疗法与中医药理论渊源初探[J].中华中医药杂志,2006,21(7):434
[17]黄汉超,陈宏硅,吴伟.中医药治疗不稳定型心绞痛的临床研究与治疗进展[J].中医药学刊,2006,24(1)：101-104
[18]林木振,王孟和,刘秉忠.牡丹皮散联合阿司匹林治疗腰源性腹痛56例[J].浙江中西医结合杂志,2009,19(1)：10-11
[19]高健,段大航,蔡秀荣.大蒜油对心血管作用研究现状[J].社区医学杂志,2007,5(15)：53-55
[20]刘红杰,金若敏.薄荷油研究进展[J].山东中医药大学学报,2006,30(6):502-505
[21]杨帆,王羽梅.山鸡椒(山苍子)精油研究现状[J].韶关学院学报(自然科学),2009,30(6):80-84
[22]赵丽琴.山胡椒属萜类及其生物活性研究进展[J].时珍国医国药,2006,17(2):171-174
[23]潘年松,盛勇,张学愈.莪术抗肿瘤作用与品种和加工工艺的关系[J].中国中医药信息杂志,2006,13(8)：35-36
[24]Keita S M, Vincent C, Schmit J, et al. Efficacy of essential oil of Ocimum basilicum L. and O. gratissimum L. applied as an insecticidal fumigant and powder to control Callosobruchus maculates [J]. Journal of Stored Products Research,2001,37:339-349
[25]Kim S I, Park C, Ohh M H, et al. Contact and fumigant activities of aromatic plant extracts and essential oils against Lasioderma serricorne [J]. Journal of Stored Products Research,2003,39:11-19
[26]Aslan I, Ozbek H, Calmasur O. Toxicity of essential oil vapours to two greenhouse pests, Tetranychus urticae Koch and Bemisia tabaci Genn [J]. Industrial Crops and Products, 2004,19:167-173
[27]艾应伟,范志金,李伟,等.植物源生理活性物质与害虫防治[J].资源科学,2000,22(3)：78-81
[28]钱骅,赵伯涛,夏劲,等.九种精油的抗菌活性及其防腐特性研究[J].中国调味品,2010,35(4):69-72
[29]Valero M, Salmeron M C. Antibacterial activity of 11 essential oils against Bacillus cereus in tyndallized carrot broth [J]. International Journal of Food Microbiology,2003,85: 73-81
[30]何玮,李光武,傅佳.薰衣草及橙花精油体外抑菌作用的研究[J].安徽医药,2010,14(5)：525-527
[31]吴慧清,吴清平,石立三,等.植物精油对微生物的抑菌效果评估研究[J].食品科学,2008,29(12)：83-86
[32]马英姿,谭琴,李恒熠.樟树叶及天竺桂叶的精油抑菌活性研究[J].中南林业科技大学学报,2009,29(1)：36-40
[33]Serge A, David M. Antimicrobial properties of allicin from garlic [J]. Microbes and Infection,1999,1:125-129
[34]钟瑞敏,肖仔君,张振明,等.小茴香籽精油成分及其抗菌活性研究[J].林产化学与工业,2007,27(6)：36-40
[35]陈丽艳,崔志恒.植物精油抗菌活性的研究进展[J].黑龙江医药,2006,19(3):197-198
[36]Hazzit M, Baaliouamer A, Faleiro M L, et al. Composition of the essential oils of Thymus and Origanum species from Algeria and their antioxidant and antimicrobial activities [J]. Journal of Agricultural and Food Chemistry,2006,54:6314-6321
[37]胡艳芬,江文斌,蒋琳琳,等.百里香酚和香芹酚体外抑菌作用的研究[J].兽药与饲料,2010,10:107
[38]楼兴隆,李晓明,张鞍灵,等.丁香油抑菌成分研究[J].西北农业学报,2006,15(3):91-93
[39]吕世明,陈杖榴,陈建新,等.丁香酚体外抑菌作用研究[J].食品科学,2008,29(9):122-124
[40]郑国华,刘焱文,陈树和,等.牛至各生长期抗菌成分的分析[J].中国中药杂志,1999,24(9):536-538
[41]张静,冯岗,袁旭超,等.百里香酚抑茵活性初探[J].中国农业学报,2009,25(21)：277-280
[42]王雪梅,谌徽,李雪姣,等.天然活性单萜—柠檬烯的抑菌性能研究[J].吉林农业大学学报,2010,32(1)：24-28
[43]李京晶,籍保平,周峰,等.丁香和肉桂挥发油的提取、主要成分测定及其抗菌活性研究[J].食品科学,2006,27(8):64-68
[44]钱昆,周涛.植物源天然防腐剂应用和机理研究的最新进展[J].中国食品添加剂,2006,5：100-104
[45]NCCLS. Methods for Dilution Antimicrobial Susceptibility Tests for Bacterict That Grow Aerobically: Approved Standard-Sixth Edition [M]. National Committee for Clinical Laboratory Standards,2002
[46]Reza S, Rahman A, Lee J, et al. Potential roles of essential oil and organic extracts of Zizyphus jujube in inhibiting food-borne pathogens [J]. Food Chemistry,2010,119: 981-986
[47]Davidson P M, Parish M E.. Methods for testing the efficacy of food antimicrobials [J]. Food Technology,1989,43:148-155
[48]李宏.精油的抗病毒和抗微生物作用[J].香料香精化妆品,2002,5:5-8
[49]王帆,杨静东,王春梅,等.复配植物源杀菌剂的开发研究[J].江西农业学报,2010,22(2):87-89
[50]顾仁勇,傅伟昌,银永忠.丁香和肉桂精油联合抗菌作用初步研究[J].食品科学,2008,29(10)：115-119
[51]吕飞.生物抗菌包装体系及其对黑鱼品质影响的研究[D].浙江：浙江大学博士学位论文,2009
[52]Yamazaki K, Yamamoto T, Kawai Y, et al. Enhancement of antilisterial activity of essential oil constituents by nisin and diglycerol fatty acid ester [J]. Food microbiology, 2004,21:283-289
[53]Turgis M, Han J, Caillet S, et al.. Antimicrobial activity of mustard essential oil against Escherichia coli O157:H7 and Salmonella typhi [J]. Food control,2009,20:1073-1079
[54]Oussalah M, Caillet S, Saucier L, et al. Antimicrobial effects of selected plant essential oils on the growth of a Pseudomonas putida strain isolated from meat [J]. Meat Science, 2006,73:236-244
[55]孙峋汪,汪靖超,李洪涛,等.迷迭香酸的抗菌机理研究[J].青岛大学学报(自然科学版),2005,18(4)：41-45
[56]Pasqua R D, Betts G, Hoskins N, et al. Membrane toxicity of antimicrobial compounds from essential oils [J]. Journal of Agricultural and Food Chemistry,2007,55:4863-4870
[57]郑罡,曹煜,魏羽佳.茶树油对白假丝酵母菌抑菌效果[J].中国消毒学杂志,2007,24(5):438440
[58]顾仁勇.五种香辛料精油的提取及用于冷却猪肉保鲜研究[D].湖南：湖南农业大学硕士学位论文,2007
[59]李鹏霞,张兴,刘亚敏,等.36种精油对采后番茄防腐保鲜活性的影响[J].西北林学院学报,2008,23(3)：156-159
[60]刘芳,张小青,吴三林,等.峨眉含笑精油对冬枣保鲜的研究[J].长春师范学院学报(自然科学版),2008,27(3)：65-69
[61]张春红,刘静,陈丽,等.三种植物精油对霉菌的抑制效果测定及其在大米储藏中应用研究[J].食品科技,2007,32(12):105-108
[62]Shelef L A, Jyothi E K, Bulgarelli M A. Growth of enteropathogenic and spoilage bacteria in sage-containing broth and foods [J]. Journal of Food Science,1984,49:737-740
[63]疏秀林,施庆珊,欧阳友生,等.植物精油的抗菌特性及在食品工业中应用研究新进展[J].生物技术,2006,16(6):89-92
[64]Stammati A, Bonsi P, Zucco F, et al. Toxicity of selected plant volatiles in microbial and mammalian short-term assays [J]. Food and chemical Toxiology,1999(37):813-823
[65]Celiktas O, Kocabas E, Bedir E, et al. Antimicrobial activities of methanol extracts and essential oils of Rosmarinus oficinalis, depending on location and seasonal variations [J]. Food Chemistry,2007,100:553-559
[66]Jacob C, Mathiasen L, Powell D. Designing effective messages for microbial food safety hazards [J]. Food Control,2010,21:1-6
[67]Fleming-Jones M, Smith R. Volatile organic compounds in foods: a five year study [J]. Journal of Agricultural and Food Chemistry,2003,51:8120-8127
[68]Goni P, Lopez P, Sanchez C, et al. Antimicrobial activity in the vapour phase of a combination of cinnamon and clove essential oils [J]. Food Chemistry,2009,116:982-989
[69]Hsieh P C, Mau J L, Huang S H. Antimicrobial effect of various combinations of plant extracts [J]. Food Microbiology,2001,18:35-43
[70]Nazer A I, Kobilinsky A, Tholozan J L, et al. Combinations of food antimicrobials at low levels to inhibit the growth of Salmonella sv. Typhimurium:A synergistic effect? [J]. Food Microbiology,2005,22:391-398
[71]Gutierrez J, Barry-Ryan C, Bourke P. Antimicrobial activity of plant essential oils using food model media: Efficacy, synergistic potential and interactions with food components [J]. Food Microbiology,2009,26:142-150
[72]Firuzi O, Asadollahi M, Gholami M, et al. Composition and biological activities of essential oils from four Heracleum species [J]. Food Chemistry,2010,122:117-122
[73]Rota M C, Carraminana J J, Burillo J, et al. In vitro antimicrobial activity of essential oils from aromatic plants against selected foodborne pathogens [J]. Journal of Food Protection, 2004,67:1252-1256
[74]Weerakkody N S, Caffin N, Turner M S, et al. In vitro antimicrobial activity of less-utilized spice and herb extracts against selected food-borne bacteria [J]. Food Control, 2010,21:1408-1414
[75]Hemaiswarya S, Kruthiventi A K, Doble M. Synergism between natural products and antibiotics against infectious diseases [J]. Phytomedicine,2008,15:639-652
[76]Rota M C, Herrera A, Martinez R M, et al. Antimicrobial activity and chemical composition of Thymus vulgaris, Thymus zygis and Thymus hyemalis essential oils [J]. Food Control,2008,19:681-687
[77]Oussalah M, Caillet S, Saucier L, et al. Antimicrobial effects of selected plant essential oils on the growth of a Pseudomonas putida strain isolated from meat [J]. Meat Science, 2006,73:236-244
[78]Vaara, M. Agents that increase the permeability of the outer membrane [J]. Microbiological Reviews,1992,56:395-411
[79]Elgayyar M, Draughon F A, Golden D A, et al. Antimicrobial activity of essential oils from plants against selected pathogenic and saprophytic microorganisms [J]. Journal of Food Protection,2001,64:1019-1024
[80]Burt S, Reinders R D. Antibacterial activity of selected plant essential oils against Escherichia coli O157.H7 [J]. Applied Microbiology,2003,36:162-167
[81]Gutierrez J, Rodriguez G, Barry-Ryan C, et al. Efficacy of plant essential oils against food-borne pathogens and spoilage bacteria associated with ready to eat vegetables: antimicrobial and sensory screening [J]. Journal of Food Protection,2008,71:1846-1854
[82]Hammer K A, Carson C F, Riley T V. Antimicrobial activity of essential oils and other plant extracts [J]. Journal of Applied Microbiology,1999,86:985-990
[83]Runyoro D, Ngassapa O, Vagionas K, et al. Chemical composition and antimicrobial activity of the essential oils of four Ocimum species growing in Tanzania [J]. Food Chemistry,2010,119:311-316
[84]Grayer R J, Kite G C, Goldstone F J, et al. Infraspecific taxonomy and essential oil chemotypes in sweet basil, Ocimum basilicum[J]. Phytochemistry,1996,43:1033-1039
[85]Friedman M, Henika P R, Levin C E, et al. Antibacterial activities of plant essential oils and their components against escherichia coli O157.H7 and salmonella enterica in Apple Juice [J]. Journal of Agricultural and Food Chemistry,2004,52:6042-6048
[86]Yamamoto H, Ogawa T. Antimicrobial activity of perilla seed polyphenols against oral pathogenic bacteria [J]. Bioscience, Biotechnology, and Biochemistry,2002,66:921-924
[87]Dorman H, Deans S G. Antimicrobial agents from plants: antibacterial activity of plant volatile oils [J]. Journal of Applied Microbiology,2000,88:308-316
[88]Gill A O, Delaquis P, Russo P, et al. Evaluation of antilisterial action of cilantro oil on vacuum packed ham [J]. International Journal of Food Microbiology,2002,73:83-92
[89]Burt S. Essential oils: Their antibacterial properties and potential applications in foods-A review [J]. International Journal of Food Microbiology,2004,94:223-253
[90]Ultee A, Kets E, Alberda M, et al. Adaptation of the food-borne pathogen Bacillus cereus to carvacrol [J]. Archives of Microbiology,2000,174:233-238
[91]Naigre R, Kalck P, Roques C, et al. Comparison of antimicrobial properties of monoterpenes and their carbonylated products [J]. Planta Medica,1996,62:275-277
[92]Uhart M, Maks N, Ravishankar S. Effect of spices on growth and survival of Salmonella typhimurium DT 104 in ground beef stored at 4 and 8℃[J]. Journal of Food Safety,2006, 26:115-125
[93]Devlieghere F, Vermeulen A, Debevere J. Chitosan: antimicrobial activity, interactions with food components and applicability as a coating on fruit and vegetables [J]. Food Microbiology,2004,21:703-714
[94]Adams R P. Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy [M]. Allured Publishing Corporation:USA,2001
[95]Panizzi L, Flamini G, Cioni P L, et al. Composition and antimicrobial properties of essential oils of four Mediterranean Lamiaceae [J]. Journal of Ethnopharmacology,1993, 39:167-170
[96]Ceylan E, Fung D Y C. Antimicrobial activity of spices [J]. Journal of Rapid Methods and Automation in Microbiology,2004,12:1-55
[97]Bozen B, Dukic N M, Simin N, et al. Characterization of the volatile composition of essential oils of some Lamiaceae spices and the antimicrobial and antioxidant activities of the entire oils [J]. Journal of Agricultural and Food Chemistry,2006,54:1822-1828
[98]Vina A, Murillo E. Essential oil composition from twelve varieties of basil (Ocimum spp) grown in Colombia [J]. Journal of the Brazilian Chemical Society,2003,14:744-749
[99]Fang T, Goto M, Sasaki M, et al. Combination of supercritical CO2 and vacuum distillation for the fractionation of bergamot oil [J]. Journal of Agricultural and Food Chemistry,2004,52:5162-5167
[100]Figoli A, Donato L, Carnevale R, et al. Bergamot essential oil extraction by pervaporation [J]. Desalination,2006,193:160-165
[101]Nitta M, Kobayashi H, Ohnishi-Kameyama M, et al. Essential oil variation of cultivated and wild perilla analyzed by GC/MS [J]. Biochemical Systematics and Ecology,2006,34: 25-37
[102]Calvey E M, White K D, Matusil J E, et al. Allium chemistry: identification of organosulfur compounds in ramp (Allium tricoccum) homogenates [J]. Phytochemistry, 1998,49:359-364
[103]吴玉荷,杨小华,蒋灵芝.薇甘菊水提物的抑菌试验研究[J].中国野生植物资源,2007,26(1)：51-51
[104]谢家仪,董光军,刘振英.扫描电镜的微生物样品制备方法[J].电子显微学报,2005,24(4):440
[105]于庆华,殷文政,艾启俊,等.鹿蹄草素对金黄色葡萄球菌和绿脓杆菌的抑制效果观察[J].食品科学,2008,29(4):90-93
[106]杜春明,焦振泉,郭云昌,等.唐菖蒲伯克霍尔德菌菌体脂肪酸成分测定与分析[J].卫生研究,2005,34(5):613-616
[107]王关林,蒋丹,方宏筠.天南星的抑菌作用及其机理研究[J].畜牧兽医学报,2004,35(30)：280-285
[108]Tajkarimi M M, Ibrahima S A, Cliver D O. Antimicrobial herb and spice compounds in food [J]. Food Control,2010,21:1199-1218
[109]Sikkema J, De Bont J A M, Poolman B. Mechanisms of membrane toxicity of hydrocarbons [J]. Microbiological Reviews,1995,59:201-222
[110]Lambert R J W, Skandamis P N, Coote P, et al. A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol [J]. Journal of Applied Microbiology,2001,91:453-462
[111]Carson C F, Mee B J, Riley T V. Mechanism of action of Melaleuca alternifolia (tea tree) oil on Staphylococcus aureus determined by time-kill, lysis, leakage and salt tolerance assays and electron microscopy [J]. Antimicrobial Agents and Chemotherapy,2002,46: 1914-1920
[112]Moreira M R, Ponce A G, Del Valle C E, et al. Inhibitory parameters of essential oils to reduce a foodborne pathogen [J]. LWT-Food Science and Technology,2005,38:565-570
[113]Santiesteban-Lopez A, Palou E, Lopez-Malo A. Susceptibility of food-borne bacteria to binary combinations of antimicrobials at selected aw and pH [J]. Journal of Applied Microbiology,2007,102:486-497