بررسی ترکیبات اسانس و اثر ضدباکتریایی عصاره‌های آبی و متانولی گل میمونی اسکروفولاریا خراسانیکا بر برخی از پاتوژن‌های مهم غذایی در شرایط آزمایشگاهی

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی ‌ارشد، گروه علوم و صنایع غذایی، واحد قوچان، دانشگاه آزاد اسلامی، قوچان، ایران

2 باشگاه پژوهشگران جوان و نخبگان، واحد قوچان،‌ دانشگاه آزاد اسلامی، قوچان، ایران

3 استادیار گروه علوم و صنایع غذایی، دانشکده کشاورزی دانشگاه فردوسی مشهد، مشهد، ایران

چکیده

گیاه اسکروفولاریا خراسانیکا (khorassanicaScrophularia)از تیره گل میمون و گونه بومی ایران و خراسان است که اطلاعات کافی در مورد آنالیز اسانس و اثر ضد­­­­­­باکتریایی آن موجود نیست. با توجه به استفاده‌های درمانی محلی این گیاه و سایر گونه‌های آن، در این مطالعه ترکیبات اسانس و اثر عصاره‌های آبی و متانولی گیاه مذکور بر باکتری‌های اشریشیاکولایو استافیلوکوکوس اورئوس مورد بررسی قرار گرفت. عصاره‌ها به روش ماسراسیون و اسانس توسط تقطیر با آب تهیه گردیدند. ترکیب اسانس توسط GC/MS و حداقل غلظت مهاری (MIC) و حداقل غلظت کشندگی(MBC) با روش میکرودایلوشن براث اندازه‌گیری شد. در آنالیز اسانس 28 ترکیب شناسایی شد که در آن اجزای اصلی شامل 3،1- بیس(تری متیل سیلیل) بنزن (93/34%)، پاراسیمن (55/12%)، پالمتیک اسید (79/6%)، تیمول (22/6%)، لینالول (59/4%)، سوربیتول (87/1%)، کارواکرول (74/1%)، گاماترپینن (91/1%)، بتا-المن (03/1 %) و آلفاپینن (3/0 %) بود. نتایج نشان داد عصاره آبی گیاه دارای اثر ضدباکتری قوی‌تری نسبت به عصاره متانولی بود. همچنین هر دو عصاره آبی و متانولی اسکروفولاریا خراسانیکا دارای اثر بازدارندگی و کشندگی در غلظت‌های مورد بررسی بودند. حداقل غلظت بازدارندگی عصاره آبی علیه اشریشیا کولای و استافیلوکوکوس اورئوس به‌ترتیب 25/6 و mg/ml 5/12 و برای عصاره متانولی به ترتیب 25 و mg/ml  5/12 تعیین شد. لذا می‌توان از این عصاره به‌عنوان عوامل ضدمیکروبی طبیعی استفاده نمود.

کلیدواژه‌ها


عنوان مقاله [English]

In vitro study on the composition and antibacterial effects of aqueous and methanol extracts of on some of food-borne pathogens

نویسندگان [English]

  • A. Barati 1
  • A. Mohamadi Sani 2
  • M. Yavarmanesh 3
2 Young Researchers and Elite Club, Quchan Branch, Islamic Azad University, Quchan, Iran
3 Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
چکیده [English]

Scrophularia khorassanica belongs to the Scrophulariaceae family that geographically grows wild only in Iran-Khorasan. There is insufficient information about its composition and antibacterial activity. Due to the application of S. khorassanica and related species in traditional medicine, the aim of this study was to determine antimicrobial effect of S. khorassanica aqueous and methanolic extracts on the growth of Escherichia coli and Staphylococcus aureus and also to determine the composition of the essential oil in-vitro. Extracts and the essential oil were prepared respectively by maceration and hydro-distillation methods. The composition of the essential oil examined by GC/MS and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) determined by broth micro-dilution assay. Composition analysis identified a total of 28 various components in which the main components were 1,3-Bis (trimethylsilyl), benzene (34.93%), р-Cymene (12.55%), Palmitic acid (6.79), Thymol (6.22%), Linalool (4.59%), 2-undecanone(4.35%), Sorbitol(1.87%), Carvacrol (1.74%), γ-Terpinene (1.06%), β -Elemene (1.03%) and α-Pinene (0.3%). The MIC for methanolic extract against E. coli and S. aureus was 12.5 and 25 mg/ml, respectively. In the case of aqueous extract, it was estimated at 6.25 and 12.5 mg/ml, respectively. Based on these results, the aqueous extract had the strongest antibacterial effect. It was concluded that Scrophularia khorassanica extract can be used as an natural preservative.

کلیدواژه‌ها [English]

  • extract
  • MIC
  • MBC
  • Essential Oil
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● امیری، حمزه؛ لاری یزدی، حسین؛ اسماعیلی، اکبر؛ صمصام‌نیا، فرانک؛ اقبالی، داریوش؛ ویسکرمی، غلام‌حسن؛ دوستی، بهروز و نورمحمدی، احسان (1390). شناسایی ترکیب‌های تشکیل دهنده اسانس و بررسی ساختارهای ترشحی گیاه Scrophularia striata Boiss. مجله تحقیقات گیاهان دارویی و معطر ایران، سال 27، شماره 2، صفحات: 278-271.

● بهمنی، محمود؛ بهمنی، احسان؛ ممتاز، حسن و رفیعیان، محمود (1389).  مقایسه اثر گیاه گل میمونی بیابانی (Scrophularia deserti) با آمفوتریسین B بر روی کاندیدا آلبیکنس در شرایط آزمایشگاهی. مجله علمی پژوهشی دانشگاه علوم پزشکی اراک، سال 13، شماره 4، صفحات: 21-15.

● شرافتی چالشتری، فرهاد؛ شرافتی چالشتری، رضا و مومنی مریم (1387). اثر ضد میکروبی عصاره آبی و اتانولی گیاه گل میمونی (Scrophularia striata) بر اشرشیا کلی در شرایط آزمایشگاهی. مجله دانشگاه علوم پزشکی شهرکرد، ویژه‌نامه طب تکمیلی، سال 10، شماره 4، 37-32.

 ● Abbasi, N., Azizi Jaliliane, F., ABDI, M. and Saifmanesh, M. (2007). A comparative study of the antimicrobial effect of scrophularia striata boiss. Extract and selective antibiotics against Staphylococcus aureus and Pesudomonas aeroginosa, Journal of Medicinal Plants, 6(21): 10-18. [in Persian]

● Adams, R.P. (2001). Identification of Essential Oil Component by Gas Chromatography/Mass Spectroscopy. Allure Publishing crop, Illinois, P. 804.

● Akhondzade, A., Razavi, V., Misaghi, A., Abbasifar, R., Radmehr, B. and Khalighi, F. (2003). Effect of thyme essential oils on Salmonella typhimurium in brain and heart broth. Journal of Medicinal Plants. 8: 84-91.

● Amiri, H., Lari Yazdi, H., Esmaeili, A., Samsamnia, F., Eghbali, D., Viskarami, Gh., Dosti, B. and Noormohamadi, E. (Essential oil composition and anatomical study of Scrophularia striata Boiss, Iranian Journal of Medicinal and Aromatic Plants, 27(2): 278-271. [in Persian]

● Armestrong, G.L., Hollingworth, J., Morris, J.G. (1996). Emerging food borne pathogens. Escherichia coli O157:H7 as a model of entry of a new pathogen into the food supply of developed world. Epidemiology Reviews, 18: 29-51.

● Attar, F., Joharchi, M. R.,  Nowrouzi,  M. and  Hatami, A. (2006). New species of Scrophularia L. (Scrophulariaceae) from Iran. Iranian Journal of Botany, 12 (2): 193-202. Tehran.

● Bahmani, M., Bahmani, E Ghorbani, M., Momtaz, H. and Rafieian, M. (2011). The comparison of the in-vitro effects of Scrophularia deserti plant and amphotricin B on Candida albicans, Arak  Medical University Journal, 13(53): 15-21. [in Persian]

● Bahrami,  A. and Valadi, A. (2010). Effects of Scrophularia striata Ethanolic Leaves Extracts on staphylococcus aureus. International Journal of Pharmacology 6(4): 431-434.

● Bennett, M., Huang, N., Matheson, T. W., Smith,  A. K., Ittel, S., Nickerson, W. (1982). "(Hexamethylbenzene) Ruthenium Complexes".Inorganic Syntheses, 21: 74–78.

● De Martino, L., Bruno, M., Formisano, C., De Feo, V., Napolitano, F., Rosselli, S.and Senatore, F. (2009). Chemical Composition and Antimicrobial Activity of the Essential Oils from Two Species of Thymus Growing Wild in Southern Italy. Molecules 2009, 14: 4614-4624.

● Dorman, H. J. D., Deans, S. G. (2000). Antimicrobial agents from plants: antibacterial activity of plant volatile oils. Journal of Applied Microbiology, 88: 308–316.

● Etebari, M., Sajjadi, S. E., Jafarian-Dehkordi, A., Panahi, M. (2013). Antigenotoxic Effects of Methanolic and Aqueous Extracts of Kelussia odoratissima Mozaffarian against damage induced by methyl methane sulfonate. Journal of Isfahan Medical School, 30: 215.

● Fernandez, M.A., Garcia, M.D., (1996).  Antibacterial activity of the phenolic acids fractions of Scrophularia frutescens and Scrophularia sambucifolia. Journal of Ethnopharmacology, 53: 11-14.

● Frazier, W., Westhoff, D. (1987). Food Microbiology. Translated by: Mortazavi, S.A.,Kashani Neja, M., Ziaolhagh, H.(2006). 4th edition, Mashhad University Publication, pp: 531-560.

● Ghasemi Pirbalouti,  A., Bahmani, M., Avijgan,  M. (2009). Anti-Candida activity of some of the Iranian Medicinal Plants. Electronic Journal of Biology, 5(4): 85-88.

● Header, M., Henderson, M.R.H. (1994). The physicians of myddfai: the Welsh herbal tradition. Botanical Journal of Scotland, 46: 623–627.

● Helander, I.M., Alakomi, H.L., Latva Kala, K., Pol, L., von Wright, A. (1998). Characterization of the action of selected essential oil components on Gram negative bacteria. Journal of Agriculture and Food Chemistry. 46, 3590–3595.

● Javidnia,  K.,Tabatabai, M. and Shafiee, A. (1999). Volatile constituents and antimicrobial activity of Zataria multiflora, population Iran. Iranian Journal of Chemistry and Chemical Engineering, 18(1): 1-5.

● Lajimi, A.A., Barzegar, M., Tavirani, MR., Ahmadi, S., Entezari, M., Mahdavi, M. (2013). Study of antimicribial the Scrophularia striata extract. Medical Science Journal of Islamic Azad University, 23(3): 190.

● Lluis, P., Usall, J., Smilanick, J.L., Aguilar, M.J. and Vinas, I. (2002). Evaluation of food additives and low-toxicity compounds as alternative chemicals for the control of Penicillium digitatum and Penicillium italicumon citrus fruit. Pest Management Science, pp. 459-466.

● Mahboubi,  M., Kazempour, N., Boland Nazar, A.R. (2013). Total phenolic, total flavonoids, antioxidant and antimicrobial activities of Scrophularia striata Boiss Extracts. Jundishapur Journal of Natural Pharmaceutical Products. Jundishapur Journal of Natural Pharmaceutical Products, 8(1).

● Miyazawa, M., Okuno, y., Nakamura, S., Kameoka, H. (1998). Inducing activity of chemical mutagens by cinnamic acid derivatives from Scrophulia ningpoensis in the Salmonella typhimuriun TA 19535/ PSK 1002 umu test, Journal of Agricultural and Food Chemistry. 46: 904-10.

● Moreire, M.R., Ponce, A.G. and Roura, S.I. (2005). Inhibitory parameters of essential oils to reduce a food born pathogen. LWT. 38: 565- 570.

● Naeini, A., Nasseri, M., Kamali-Nejad, M., Khoshzaban, F., Rajabiyan, T., Isma'ilzadih Nami, E.x. et al. (2011). Effects of 50 essential oils and extracts medicinal plants of Iran on standard strains of Candida albicans in vitro. Journal of Medicinal Plants. 38:163-172.

● Nikolic, M., Glamoclija, J., Sokovic, M. (2014). Chemical composition, antimicrobial, antioxidant and antitumoractivity of Thymus serpyllum L., Thymus algeriensis Boiss. and Reut and Thymus vulgaris L. essential oils. Industrial Crops and Products 52,183-190.

● Oussalah, M., Caillet, S., Salmieri, S., Saucier, L., Lacroix, M. (2007). Antimicrobial effects of alginate-based films containing essential oils on Listeria monocytogenes and Salmonella typhimurium present in bologna and ham. Journal of Food Protection, 70(4):901-8.34.

● Pasdaran, A., Delazar, A., Nazemiyeh, H., Nahar, L.(2012). Chemical composition, and antibacterial (Against Staphylococcus aureus) and free-radical-scavenging activities of the essential oil of Scrophularia amplexicaulis Benth. Records of Natural Products Journal. 6:4,350 -355.

● Pasdaran, A., Nahar, L., Asnaashari, S., Sarker, S., Delazar, A. (2013). GC-MS analysis, free-radical-scavenging and insecticidal activities of essential oil of Scrophularia oxysepala Boiss. Pharmaceutical Sciences, 19(1), 1-5.

● Safavi, F., Meighani, H., Ebrahimi, P., Hafez Ghoran, S. (2012). Antioxidant and antibacterial activity of Scrophularia striata. Research in Pharmaceutical Sciences, 7(5): 852-57.

● Shareyat, S.H. (1997). Qualative and Quantitive Evaluation of the active Constituents and Control Methods for Medicinal Plants. )1997). 1st edition, Mani Publications. Esfahan, pp: 10-27.

● Sharafati-Chaleshtori, F., Shrafati-Chaleshtori, R. and Momeni, M. (2009). Comparison of the antimicrobial effects of the ethanolic and aqueous extracts of Scrophularia striata on Escherichia coli O157:H7 in vitro. Journal of Shahrekord University of Medical Sciences, 10(4): 32-37. [in Persian]

● Sikkema, J., de Bont, J.A., Poolman, B. (1994). Interactions of cyclic hydrocarbons with biological membranes. Journal of Biological Chemistry, 269(11): 8022-8028.

● Singh, N., Harvati, K., Klingenberg, P. (2012). Morphological evolution through integration: A quantitative study of cranial integration in Homo, Pan, Gorilla and Pongo. Journal of Human Evolution, 62,155-164.

● Singh, N., Singh, R.K. (2002). Efficant of inoculation and washing methods on the efficacy of different sanitizers against E.coli O157: H7 on lettuce. Food Microbiology.19: 183-193.

● Smith, S., Bannister, P., Beckmann, C., Brady, M., Clare, S., Flitney, D., Hansen, P., Jenkinson, M., Leibovici, D. and Zhang, Y. (2001). New tools for functional and structural brain image analysis. In Seventh International Conference on Functional Mapping of the Human Brain.

● Ultee, A., Kets, E.P., Alberda, M., Hoekstra, F.A., Smid, E.J. (2000). Adaptation of the food-borne pathogen Bacillus cereus to carvacrol. Archive of Microbiology, 174(4): 233-238.

● Xu, J., Zhou, F., Ji, B.P., Pei, R.S., Xu, N. (2008). The antibacterial mechanism of carvacrol and thymol against Escherichia coli. Letters in Applied Microbiology, 47(3):174-179.

● Zamanian-Azodi, M., Ardeshirylajimi, A., Ahmadi, N., Rezaee, M.B., Azizi-Jalilian, F., Khodarahmi, R. (2013). Antibacterial effects of Scrophularia striata seed aqueous extract on Staphylococcus aureus. Journal of Paramedical Sciences, 4(1): 58-63.

● Zhu, T., Xu, Y., Dong, B., Zhang, J., Wei, Z., Xu, Y., Yao, Y.(2011). Β-elemene inhibits proliferation of human glioblastoma cells through the activation of glia maturation factor β and induces sensitization to cisplatin. Oncology Reports, 26 (2): 405–13.

 

Abbasi, N., Azizi Jaliliane, F., ABDI, M. and Saifmanesh, M. (2007). A comparative study of the antimicrobial effect of scrophularia striata boiss. Extract and selective antibiotics against Staphylococcus aureus and Pesudomonas aeroginosa, Journal of Medicinal Plants, 6(21): 10-18. [in Persian]

● Adams, R.P. (2001). Identification of Essential OilComponent by Gas Chromatography/Mass Spectroscopy. Allure Publishing crop, Illinois, P. 804.

Akhondzade, A., Razavi, V., Misaghi, A., Abbasifar, R., Radmehr, B. and Khalighi, F. (2003). Effect of thyme essential oils on Salmonella typhimurium in brain and heart broth. Journal of Medicinal Plants. 8: 84-91.

Amiri, H., Lari Yazdi, H., Esmaeili, A., Samsamnia, F., Eghbali, D., Viskarami, Gh., Dosti, B. and Noormohamadi, E. (Essential oil composition and anatomical study of Scrophularia striata Boiss, Iranian Journal of Medicinal and Aromatic Plants, 27(2): 278-271. [in Persian]

● Armestrong, G.L., Hollingworth, J., Morris, J.G. (1996). Emerging food borne pathogens. Escherichia coli O157:H7 as a model of entry of a new pathogen into the food supply of developed world. Epidemiology Reviews, 18: 29-51.

● Attar, F., Joharchi, M. R.,  Nowrouzi,  M. and  Hatami, A. (2006). New species of Scrophularia L. (Scrophulariaceae) from Iran. Iranian Journal of Botany, 12 (2): 193-202. Tehran.

Bahmani, M., Bahmani, E Ghorbani, M., Momtaz, H. and Rafieian, M. (2011). The comparison of the in-vitro effects of Scrophularia deserti plant and amphotricin B on Candida albicans, Arak  Medical University Journal, 13(53): 15-21. [in Persian]

● Bahrami,  A. and Valadi, A. (2010). Effects of Scrophularia striata Ethanolic Leaves Extracts on staphylococcus aureus. International Journal of Pharmacology 6(4): 431-434.

Bennett, M., Huang, N., Matheson, T. W., Smith,  A. K., Ittel, S., Nickerson, W. (1982). "(Hexamethylbenzene) Ruthenium Complexes".Inorganic Syntheses, 21: 74–78.

De Martino, L., Bruno, M., Formisano, C., De Feo, V., Napolitano, F., Rosselli, S.and Senatore, F. (2009). Chemical Composition and Antimicrobial Activity of the Essential Oils from Two Species of Thymus Growing Wild in Southern Italy. Molecules 2009, 14: 4614-4624.

Dorman, H. J. D., Deans, S. G. (2000). Antimicrobial agents from plants: antibacterial activity of plant volatile oils. Journal of Applied Microbiology, 88: 308–316.

● Etebari, M., Sajjadi, S. E., Jafarian-Dehkordi, A., Panahi, M. (2013). Antigenotoxic Effects of Methanolic and Aqueous Extracts of Kelussia odoratissima Mozaffarian against damage induced by methyl methane sulfonate. Journal of Isfahan Medical School, 30: 215.

● Fernandez, M.A., Garcia, M.D., (1996).  Antibacterial activity of the phenolic acids fractions of Scrophularia frutescens and Scrophularia sambucifolia. Journal of Ethnopharmacology, 53: 11-14.

Frazier, W., Westhoff, D. (1987). Food Microbiology. Translated by: Mortazavi, S.A.,Kashani Neja, M., Ziaolhagh, H.(2006). 4th edition, Mashhad University Publication, pp: 531-560.

Ghasemi Pirbalouti,  A., Bahmani, M., Avijgan,  M. (2009). Anti-Candida activity of some of the Iranian Medicinal Plants. Electronic Journal of Biology, 5(4): 85-88.

Header, M., Henderson, M.R.H. (1994). The physicians of myddfai: the Welsh herbal tradition. Botanical Journal of Scotland, 46: 623–627.

● Helander, I.M., Alakomi, H.L., Latva Kala, K., Pol, L., von Wright, A. (1998). Characterization of the action of selected essential oil components on Gram negative bacteria. Journal of Agriculture and Food Chemistry. 46, 3590–3595.

● Javidnia,  K.,Tabatabai, M. and Shafiee, A. (1999). Volatile constituents and antimicrobial activity of Zataria multiflora, population Iran. Iranian Journal of Chemistry and ChemicalEngineering, 18(1): 1-5.

Lajimi, A.A., Barzegar, M., Tavirani, MR., Ahmadi, S., Entezari, M., Mahdavi, M. (2013). Study of antimicribial the Scrophularia striata extract. Medical Science Journal of Islamic Azad University, 23(3): 190.

● Lluis, P., Usall, J., Smilanick, J.L., Aguilar, M.J. and Vinas, I. (2002). Evaluation of food additives and low-toxicity compounds as alternative chemicals for the control of Penicillium digitatum and Penicillium italicumon citrus fruit. Pest Management Science, pp. 459-466.

Mahboubi,  M., Kazempour, N., Boland Nazar, A.R. (2013). Total phenolic, total flavonoids, antioxidant and antimicrobial activities of Scrophularia striata Boiss Extracts. Jundishapur Journal of Natural Pharmaceutical Products. Jundishapur Journal of Natural Pharmaceutical Products, 8(1).

● Miyazawa, M., Okuno, y., Nakamura, S., Kameoka, H. (1998). Inducing activity of chemical mutagens by cinnamic acid derivatives from Scrophulia ningpoensis in the Salmonella typhimuriun TA 19535/ PSK 1002 umu test, Journal of Agricultural and Food Chemistry. 46: 904-10.

● Moreire, M.R., Ponce, A.G. and Roura, S.I. (2005). Inhibitory parameters of essential oils to reduce a food born pathogen. LWT. 38: 565- 570.

● Naeini, A., Nasseri, M., Kamali-Nejad, M., Khoshzaban, F., Rajabiyan, T., Isma'ilzadih Nami, E.x. et al. (2011). Effects of 50 essential oils and extracts medicinal plants of Iran on standard strains of Candida albicans in vitro. Journal of Medicinal Plants. 38:163-172.

● Nikolic, M., Glamoclija, J., Sokovic, M. (2014). Chemical composition, antimicrobial, antioxidant and antitumoractivity of Thymus serpyllum L., Thymus algeriensis Boiss. and Reut and Thymus vulgaris L. essential oils. Industrial Crops and Products 52,183-190.

Oussalah, M., Caillet, S., Salmieri, S., Saucier, L., Lacroix, M. (2007). Antimicrobial effects of alginate-based films containing essential oils on Listeria monocytogenes and Salmonella typhimurium present in bologna and ham.Journal of Food Protection, 70(4):901-8.34.

Pasdaran, A., Delazar, A., Nazemiyeh, H., Nahar, L.(2012). Chemical composition, and antibacterial (Against Staphylococcus aureus) and free-radical-scavenging activities of the essential oil of Scrophularia amplexicaulis Benth. Records of Natural Products Journal. 6:4,350 -355.

Pasdaran, A., Nahar, L., Asnaashari, S., Sarker, S., Delazar, A. (2013). GC-MS analysis, free-radical-scavenging and insecticidal activities of essential oil of Scrophularia oxysepala Boiss. Pharmaceutical Sciences, 19(1), 1-5.

Safavi, F., Meighani, H., Ebrahimi, P., Hafez Ghoran, S. (2012). Antioxidant and antibacterial activity of Scrophularia striata. Research in Pharmaceutical Sciences, 7(5): 852-57.

Shareyat, S.H. (1997). Qualative and Quantitive Evaluation of the active Constituents and Control Methods for Medicinal Plants. )1997). 1st edition, Mani Publications. Esfahan,pp: 10-27.

Sharafati-Chaleshtori, F., Shrafati-Chaleshtori, R. and Momeni, M. (2009). Comparison of the antimicrobial effects of the ethanolic and aqueous extracts of Scrophularia striata on Escherichia coli O157:H7 in vitro. Journal of Shahrekord University of Medical Sciences, 10(4): 32-37. [in Persian]

Sikkema, J., de Bont, J.A., Poolman, B. (1994). Interactions of cyclic hydrocarbons with biological membranes.Journal of Biological Chemistry, 269(11): 8022-8028.

● Singh, N., Harvati, K., Klingenberg, P. (2012). Morphological evolution through integration: A quantitative study of cranial integration in Homo, Pan, Gorilla and Pongo. Journal of Human Evolution, 62,155-164.

Singh, N., Singh, R.K. (2002). Efficant of inoculation and washing methods on theefficacy of different sanitizers againstE.coli O157: H7 on lettuce. FoodMicrobiology.19: 183-193.

● Smith, S., Bannister, P., Beckmann, C., Brady, M., Clare, S., Flitney, D., Hansen, P., Jenkinson, M., Leibovici, D. and Zhang, Y. (2001). New tools for functional and structural brain image analysis. In Seventh International Conference on Functional Mapping of the Human Brain.

Ultee, A., Kets, E.P., Alberda, M., Hoekstra, F.A., Smid, E.J. (2000). Adaptation of the food-borne pathogen Bacillus cereus to carvacrol.Archive of Microbiology, 174(4): 233-238.

Xu, J., Zhou, F., Ji, B.P., Pei, R.S., Xu, N. (2008). The antibacterial mechanism of carvacrol and thymol against Escherichia coli. Letters in Applied Microbiology, 47(3):174-179.

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