Антимикробная активность эфирных масел некоторых растений Казахстана
Ключевые слова:
эфирные масла, антимикробная активность, Staphylococcus aureus, Escherichia coli, Candida albicans,Аннотация
Определена антимикробная активность 15 образцов эфирных масел, выделенных из 9 видов растений Казахстана: Artemisia kotuchovii Kupr., Artemisia scoparia Waldst. et Kit., Artemisia absinthium L., Ferula iliensis Krasn. ex Korovin, Ferula ovina (Boiss.) Boiss., Heracleum dissectum Ldb., Angelica decurrens (Ledeb.) B. Fedtsch., Thymus marschallianus Willd. и Juniperus sabina L. Эфирные масла были получены из отдельных частей растений на разных фазах вегетации. Наибольший ингибирующий эффект в отношении всех исследованных штаммов микроорганизмов имело эфирное масло Thymus marschallianus, которое в концентрации 0,31 мкг/мл полностью ингибировало рост Staphylococcus aureus, Escherichia coli и Candida albicans. Высокой антимикробной активностью также обладали эфирные масла из корней эндемичного вида Ferula iliensis: в концентрации 0,31 мкг/мл эти эфирные масла полностью подавляли рост Candida albicans, а в концентрации 2,5 мкг/мл ингибировали рост Staphylococcus aureus и Escherichia coli. Эфирное масло, выделенное из цветков и листьев полыни Котухова, в концентрации 1,25 мкг/мл подавляло рост всех исследуемых нами штаммов микроорганизмов. Это первое исследование, демонстрирующее антимикробную активность эндемичных видов Казахстана – Ferula iliensis и Artemisia kotuchovii.
Библиографические ссылки
Литература
1. Mancini E., Senatore F., Del Monte D., De Martino L., Grulova D., Scognamiglio M., Snoussi M., De Feo V. Studies on chemical composition, antimicrobial and antioxidant activities of five Thymus vulgaris L. essential oils // Molecules. – 2015. – Vol. 20, No. 7. – P. 12016-12028. DOI: 10.3390/molecules200712016.
2. Melo J.O., Fachin A.L., Rizo W.F., Jesus H.C.R., Arrigoni-Blank M.F., Alves P.B., Marins M.A., França S.C., Blank A.F. Cytotoxic effects of essential oils from three Lippia gracilis Schauer genotypes on HeLa, B16, and MCF-7 cells and normal human fibroblasts // Genetics and Molecular Research. – 2014. – Vol. 13, No. 2. – P. 2691-2697. DOI: 10.4238/2014.April.8.12.
3. Seema F., Vijaya P.P, Manivannan V. Immunomodulatory activity of geranial, geranial acetate, gingerol, and eugenol essential oils: evidence for humoral and cell-mediated responses // Avicenna J Phytomed. – 2013. – Vol. 3, No. 3. – P. 224-230.
4. Chao S., Young G., Oberg C., Nakaoka K. Inhibition of methicillin-resistant Staphylococcus aureus (MRSA) by essential oils // Flavour Fragr. J. – 2008. – Vol. 23, No. 6. – P. 444-449. DOI: 10.1002/ffj.1904.
5. Tohidpour A., Sattari M., Omidbaigi R., Yadegar A., Nazemi J. Antibacterial effect of essential oils from two medicinal plants against methicillin-resistant Staphylococcus aureus (MRSA) // Phytomedicine. – 2010. – Vol. 17. – P. 142–145. DOI: 10.1016/j.phymed.2009.05.007.
6. Juliani H.R., Biurrun F., Koroch A.R., Oliva M.M., Demo M.S., Trippi V.S., Zygadlo J.A. Chemical constituents and antimicrobial activity of the essential oil of Lantana xenica mold // Planta Med. – 2002. – Vol. 68. – P. 756-762. DOI: 10.1055/s-2002-33803.
7. Торина А.К., Бисенова Г.Н., Шегебаева А.А., Ибраева А., Рязанцев О., Атажанова Г.А. Антимикробная активность основных компонентов эфирных масел и их некоторых производных // Вестник науки Казахского Агротехнического университета им. С. Сейфуллина. – 2014. – №3. – С.54-62.
8. Паршина Г. Н., Айткельдиева С.А., Мукиянова У.С., Бейсетбаева Г.М. Содержание эфирных масел и их антимикробная активность в лекарственных растениях семейства Lamiaceae Lindl., культивируемых в Акмолинской области // Известия Национальной академии наук Республики Казахстан. Серия биологическая и медицинская. – 2014. – № 2. – С. 19-24.
9. Bassole I.H.N., Lamien-Meda A., Bayala B., Tirogo S., Franz C., Novak J., Nebie R.C., Dicko M.H. Composition and antimicrobial activities of Lippia multiflora Moldenke, Mentha x piperita L. and Ocimum basilicum L. essential oils and their major monoterpene alcohols alone and in combination // Molecules. – 2010. – Vol. 10. – P. 7825-7839. DOI: 10.3390/molecules15117825.
10. Bassole I.H.N., Juliani H. R. Essential oils in combination and their antimicrobial properties // Molecules. – 2012. – Vol. 17. – P. 3989-4006. DOI: 10.3390/molecules17043989.
11. Burt S. Essential oils: their antimicrobial properties and potential applications in foods – a review // Int. J. Food Microbiol. – 2004. – Vol. 94. – P. 223-253. DOI: org/10.1016/j.ijfoodmicro.2004.03.022.
12. Koroch A., Juliani H.R., Zygadlo J.A. Bioactivity of essential oils and their components // Flavours and Fragrances Chemistry, Bioprocessing and sustainability, Berger, R.G., Ed.; Springer Verlag: Berlin, Germany. – 2007. – P. 87-115. ISBN 978-3-540-49338-9.
13. Delaquis P.J., Stanish K., Girard B, Mazza G. Antimicrobial activity of individual and mixed fractions of dill, cilantro, coriander and eucalyptus essential oils // Int. J. Food Microbiol. – 2002. – Vol. 74. – P. 101-109. DOI: 10.1016/S0168-1605(01)00734-6.
14. Котухов Ю.А., Ануфриева О.А. Artemisia kotuchovii Kupr. (Asteracea) – редкий исчезающий вид флоры Казахстана // Ботанические исследования Сибири и Казахстана. – 2016. – № 22. – С. 60-70.
15. Красная книга Казахстана. – Изд. 2-е, переработанное и дополненное. Т. 2: Растения. – Астана, ТОО «АртPrintXXI». – 2014. – С. 222. ISBN 978-601-80334-7-6.
16. Murray P.R., Baron E.J., Pfaller M.A., Tenover F.C., Yolke R.H. Manual of clinical microbiology. 7th ed. – Washington: ASM, 1995. – P. 1773. ISBN 000-1555811264.
17. Kavoosi G., Rowshan V. Chemical composition and antimicrobial activities of essential oil obtained from Ferula assa-foetida oleo-gum-resin: Effect of collection time // Food Chemistry. – 2013. – Vol. 138. – P. 2180-2187. DOI: 10.1016/j.foodchem.2012.11.131.
18. Schepetkin I.A., Kushnarenko S.V., Özek G., Kirpotina L.N., Utegenova G.A., Kotukhov Y.A., Danilova A.N., Özek T., Başer K.H.C., Quinn M.T. Inhibition of human neutrophil responses by essential oil of Artemisia kotuchovii and its constituents – J. Agric. Food Chem. – 2015. – Vol. 63. – P. 4999-5007. DOI: 10.1021/acs.jafc.5b01307.
References
1. Mancini E, Senatore F, Del Monte D, De Martino L, Grulova D, Scognamiglio M, Snoussi M, De Feo V (2015) Studies on chemical composition, antimicrobial and antioxidant activities of five Thymus vulgaris L. essential oils. Molecules, 20:12016-12028. DOI: 10.3390/molecules200712016.
2. Melo JO, Fachin AL, Rizo WF, Jesus HCR, Arrigoni-Blank MF, Alves PB, Marins MA, França SC, Blank AF (2014) Cytotoxic effects of essential oils from three Lippia gracilis Schauer genotypes on HeLa, B16, and MCF-7 cells and normal human fibroblasts. Genetics and Molecular Research, 13:2691-2697. DOI: 10.4238/2014.April.8.12.
3. Seema F, Vijaya PP, Manivannan V (2013) Immunomodulatory activity of geranial, geranial acetate, gingerol, and eugenol essential oils: evidence for humoral and cell-mediated responses. Avicenna J Phytomed, 3:224-230.
4. Chao S, Young G, Oberg C, Nakaoka K (2008) Inhibition of methicillin-resistant Staphylococcus aureus (MRSA) by essential oils. Flavour Fragr. J, 23:444-449. DOI: 10.1002/ffj.1904.
5. Tohidpour A, Sattari M, Omidbaigi R, Yadegar A, Nazemi J (2010) Antibacterial effect of essential oils from two medicinal plants against methicillin-resistant Staphylococcus aureus (MRSA). Phytomedicine, 17:142–145. DOI: 10.1016/j.phymed.2009.05.007.
6. Juliani HR, Biurrun F, Koroch AR, Oliva MM, Demo MS, Trippi VS, Zygadlo JA (2002) Chemical constituents and antimicrobial activity of the essential oil of Lantana xenica mold. Planta Med, 68:756-762. DOI: 10.1055/s-2002-33803.
7. Torina AK, Bisenova GN, Shegebaeva AA, Ibraeva A, Rjazancev O, Atazhanova GA (2014) Antimicrobial activity of main constituents of essential oils and some their derivatives. Bulletin of Science of S. Seifullin Kazakh Agrotechnical University [Antimikrobnaja aktivnost' osnovnyh komponentov jefirnyh masel i ih nekotoryh proizvodnyh. Vestnik nauki Kazahskogo Agrotehnicheskogo universiteta im. S. Sejfullina] 3:54-62. (In Russian).
8. Parshina GN, Ajtkel'dieva SA, Mukijanova US, Bejsetbaeva GM (2014) Content of essential oils and their antimicrobial activity in medicinal plants from Lamiaceae Lindl. Family cultivated in Akmola region. Bulletin of National Academy of Sciences of Republic of Kazakhstan. Biological and Medical Series [Soderzhanie jefirnyh masel i ih antimikrobnaja aktivnost' v lekarstvennyh rastenijah semejstva Lamiaceae Lindl., kul'tiviruemyh v Akmolinskoj oblasti. Izvestija Nacional'noj akademii nauk Respubliki Kazahstan. Serija biologicheskaja i medicinskaja]. 2:19-24. (In Russian).
9. Bassole IHN, Lamien-Meda A, Bayala B, Tirogo S, Franz C, Novak J, Nebie RC, Dicko MH (2010) Composition and antimicrobial activities of Lippia multiflora Moldenke, Mentha x piperita L. and Ocimum basilicum L. essential oils and their major monoterpene alcohols alone and in combination. Molecules,10:7825-7839. DOI: 10.3390/molecules15117825.
10. Bassole IHN, Juliani HR (2012) Essential oils in combination and their antimicrobial properties. Molecules, 17:3989-4006. DOI: 10.3390/molecules17043989.
11. Burt S (2004) Essential oils: their antimicrobial properties and potential applications in foods – a review. Int. J. Food Microbiol, 94:223-253. DOI: org/10.1016/j.ijfoodmicro.2004.03.022.
12. Koroch A, Juliani HR, Zygadlo JA (2007) Bioactivity of essential oils and their components. Flavours and Fragrances Chemistry, Bioprocessing and sustainability, Berger, R.G., Ed.; Springer Verlag: Berlin, Germany, 87-115. ISBN 978-3-540-49338-9.
13. Delaquis PJ, Stanish K, Girard B, Mazza G (2002) Antimicrobial activity of individual and mixed fractions of dill, cilantro, coriander and eucalyptus essential oils. Int. J. Food Microbiol, 74:101-109. DOI: 10.1016/S0168-1605(01)00734-6.
14. Kotuhov JuA, Anufrieva OA (2016) Artemisia kotuchovii Kupr. (Asteracea) – rare endangered species of Kazakhstan flora. Botanical Research in Siberia and Kazakhstan [Artemisia kotuchovii Kupr. (Asteracea) – redkij ischezajushhij vid flory Kazahstana. Botanicheskie issledovanija Sibiri i Kazahstana] 22:60-70 (In Russian).
15. Red Data Book of Kazakhstan (2014) The 2nd edition revised and supplemented. Vol. 2: Plants. Аstana, LTD «АpтPrintXXI», 222. ISBN 978-601-80334-7-6.
16. Murray P.R., Baron E.J., Pfaller M.A., Tenover F.C., Yolke R.H. Manual of clinical microbiology. 7th ed. – Washington: ASM, 1995. – P. 1773. ISBN 000-1555811264.
17. Kavoosi G, Rowshan V (2013) Chemical composition and antimicrobial activities of essential oil obtained from Ferula assa-foetida oleo-gum-resin: Effect of collection time. Food Chemistry, 138:2180-2187. DOI: 10.1016/j.foodchem.2012.11.131.
18. Schepetkin IA, Kushnarenko SV, Özek G, Kirpotina LN, Utegenova GA, Kotukhov YA, Danilova AN, Özek T, Başer KHC, Quinn MT (2015) Inhibition of human neutrophil responses by essential oil of Artemisia kotuchovii and its constituents. J. Agric. Food Chem, 63:4999-5007. DOI: 10.1021/acs.jafc.5b01307.
1. Mancini E., Senatore F., Del Monte D., De Martino L., Grulova D., Scognamiglio M., Snoussi M., De Feo V. Studies on chemical composition, antimicrobial and antioxidant activities of five Thymus vulgaris L. essential oils // Molecules. – 2015. – Vol. 20, No. 7. – P. 12016-12028. DOI: 10.3390/molecules200712016.
2. Melo J.O., Fachin A.L., Rizo W.F., Jesus H.C.R., Arrigoni-Blank M.F., Alves P.B., Marins M.A., França S.C., Blank A.F. Cytotoxic effects of essential oils from three Lippia gracilis Schauer genotypes on HeLa, B16, and MCF-7 cells and normal human fibroblasts // Genetics and Molecular Research. – 2014. – Vol. 13, No. 2. – P. 2691-2697. DOI: 10.4238/2014.April.8.12.
3. Seema F., Vijaya P.P, Manivannan V. Immunomodulatory activity of geranial, geranial acetate, gingerol, and eugenol essential oils: evidence for humoral and cell-mediated responses // Avicenna J Phytomed. – 2013. – Vol. 3, No. 3. – P. 224-230.
4. Chao S., Young G., Oberg C., Nakaoka K. Inhibition of methicillin-resistant Staphylococcus aureus (MRSA) by essential oils // Flavour Fragr. J. – 2008. – Vol. 23, No. 6. – P. 444-449. DOI: 10.1002/ffj.1904.
5. Tohidpour A., Sattari M., Omidbaigi R., Yadegar A., Nazemi J. Antibacterial effect of essential oils from two medicinal plants against methicillin-resistant Staphylococcus aureus (MRSA) // Phytomedicine. – 2010. – Vol. 17. – P. 142–145. DOI: 10.1016/j.phymed.2009.05.007.
6. Juliani H.R., Biurrun F., Koroch A.R., Oliva M.M., Demo M.S., Trippi V.S., Zygadlo J.A. Chemical constituents and antimicrobial activity of the essential oil of Lantana xenica mold // Planta Med. – 2002. – Vol. 68. – P. 756-762. DOI: 10.1055/s-2002-33803.
7. Торина А.К., Бисенова Г.Н., Шегебаева А.А., Ибраева А., Рязанцев О., Атажанова Г.А. Антимикробная активность основных компонентов эфирных масел и их некоторых производных // Вестник науки Казахского Агротехнического университета им. С. Сейфуллина. – 2014. – №3. – С.54-62.
8. Паршина Г. Н., Айткельдиева С.А., Мукиянова У.С., Бейсетбаева Г.М. Содержание эфирных масел и их антимикробная активность в лекарственных растениях семейства Lamiaceae Lindl., культивируемых в Акмолинской области // Известия Национальной академии наук Республики Казахстан. Серия биологическая и медицинская. – 2014. – № 2. – С. 19-24.
9. Bassole I.H.N., Lamien-Meda A., Bayala B., Tirogo S., Franz C., Novak J., Nebie R.C., Dicko M.H. Composition and antimicrobial activities of Lippia multiflora Moldenke, Mentha x piperita L. and Ocimum basilicum L. essential oils and their major monoterpene alcohols alone and in combination // Molecules. – 2010. – Vol. 10. – P. 7825-7839. DOI: 10.3390/molecules15117825.
10. Bassole I.H.N., Juliani H. R. Essential oils in combination and their antimicrobial properties // Molecules. – 2012. – Vol. 17. – P. 3989-4006. DOI: 10.3390/molecules17043989.
11. Burt S. Essential oils: their antimicrobial properties and potential applications in foods – a review // Int. J. Food Microbiol. – 2004. – Vol. 94. – P. 223-253. DOI: org/10.1016/j.ijfoodmicro.2004.03.022.
12. Koroch A., Juliani H.R., Zygadlo J.A. Bioactivity of essential oils and their components // Flavours and Fragrances Chemistry, Bioprocessing and sustainability, Berger, R.G., Ed.; Springer Verlag: Berlin, Germany. – 2007. – P. 87-115. ISBN 978-3-540-49338-9.
13. Delaquis P.J., Stanish K., Girard B, Mazza G. Antimicrobial activity of individual and mixed fractions of dill, cilantro, coriander and eucalyptus essential oils // Int. J. Food Microbiol. – 2002. – Vol. 74. – P. 101-109. DOI: 10.1016/S0168-1605(01)00734-6.
14. Котухов Ю.А., Ануфриева О.А. Artemisia kotuchovii Kupr. (Asteracea) – редкий исчезающий вид флоры Казахстана // Ботанические исследования Сибири и Казахстана. – 2016. – № 22. – С. 60-70.
15. Красная книга Казахстана. – Изд. 2-е, переработанное и дополненное. Т. 2: Растения. – Астана, ТОО «АртPrintXXI». – 2014. – С. 222. ISBN 978-601-80334-7-6.
16. Murray P.R., Baron E.J., Pfaller M.A., Tenover F.C., Yolke R.H. Manual of clinical microbiology. 7th ed. – Washington: ASM, 1995. – P. 1773. ISBN 000-1555811264.
17. Kavoosi G., Rowshan V. Chemical composition and antimicrobial activities of essential oil obtained from Ferula assa-foetida oleo-gum-resin: Effect of collection time // Food Chemistry. – 2013. – Vol. 138. – P. 2180-2187. DOI: 10.1016/j.foodchem.2012.11.131.
18. Schepetkin I.A., Kushnarenko S.V., Özek G., Kirpotina L.N., Utegenova G.A., Kotukhov Y.A., Danilova A.N., Özek T., Başer K.H.C., Quinn M.T. Inhibition of human neutrophil responses by essential oil of Artemisia kotuchovii and its constituents – J. Agric. Food Chem. – 2015. – Vol. 63. – P. 4999-5007. DOI: 10.1021/acs.jafc.5b01307.
References
1. Mancini E, Senatore F, Del Monte D, De Martino L, Grulova D, Scognamiglio M, Snoussi M, De Feo V (2015) Studies on chemical composition, antimicrobial and antioxidant activities of five Thymus vulgaris L. essential oils. Molecules, 20:12016-12028. DOI: 10.3390/molecules200712016.
2. Melo JO, Fachin AL, Rizo WF, Jesus HCR, Arrigoni-Blank MF, Alves PB, Marins MA, França SC, Blank AF (2014) Cytotoxic effects of essential oils from three Lippia gracilis Schauer genotypes on HeLa, B16, and MCF-7 cells and normal human fibroblasts. Genetics and Molecular Research, 13:2691-2697. DOI: 10.4238/2014.April.8.12.
3. Seema F, Vijaya PP, Manivannan V (2013) Immunomodulatory activity of geranial, geranial acetate, gingerol, and eugenol essential oils: evidence for humoral and cell-mediated responses. Avicenna J Phytomed, 3:224-230.
4. Chao S, Young G, Oberg C, Nakaoka K (2008) Inhibition of methicillin-resistant Staphylococcus aureus (MRSA) by essential oils. Flavour Fragr. J, 23:444-449. DOI: 10.1002/ffj.1904.
5. Tohidpour A, Sattari M, Omidbaigi R, Yadegar A, Nazemi J (2010) Antibacterial effect of essential oils from two medicinal plants against methicillin-resistant Staphylococcus aureus (MRSA). Phytomedicine, 17:142–145. DOI: 10.1016/j.phymed.2009.05.007.
6. Juliani HR, Biurrun F, Koroch AR, Oliva MM, Demo MS, Trippi VS, Zygadlo JA (2002) Chemical constituents and antimicrobial activity of the essential oil of Lantana xenica mold. Planta Med, 68:756-762. DOI: 10.1055/s-2002-33803.
7. Torina AK, Bisenova GN, Shegebaeva AA, Ibraeva A, Rjazancev O, Atazhanova GA (2014) Antimicrobial activity of main constituents of essential oils and some their derivatives. Bulletin of Science of S. Seifullin Kazakh Agrotechnical University [Antimikrobnaja aktivnost' osnovnyh komponentov jefirnyh masel i ih nekotoryh proizvodnyh. Vestnik nauki Kazahskogo Agrotehnicheskogo universiteta im. S. Sejfullina] 3:54-62. (In Russian).
8. Parshina GN, Ajtkel'dieva SA, Mukijanova US, Bejsetbaeva GM (2014) Content of essential oils and their antimicrobial activity in medicinal plants from Lamiaceae Lindl. Family cultivated in Akmola region. Bulletin of National Academy of Sciences of Republic of Kazakhstan. Biological and Medical Series [Soderzhanie jefirnyh masel i ih antimikrobnaja aktivnost' v lekarstvennyh rastenijah semejstva Lamiaceae Lindl., kul'tiviruemyh v Akmolinskoj oblasti. Izvestija Nacional'noj akademii nauk Respubliki Kazahstan. Serija biologicheskaja i medicinskaja]. 2:19-24. (In Russian).
9. Bassole IHN, Lamien-Meda A, Bayala B, Tirogo S, Franz C, Novak J, Nebie RC, Dicko MH (2010) Composition and antimicrobial activities of Lippia multiflora Moldenke, Mentha x piperita L. and Ocimum basilicum L. essential oils and their major monoterpene alcohols alone and in combination. Molecules,10:7825-7839. DOI: 10.3390/molecules15117825.
10. Bassole IHN, Juliani HR (2012) Essential oils in combination and their antimicrobial properties. Molecules, 17:3989-4006. DOI: 10.3390/molecules17043989.
11. Burt S (2004) Essential oils: their antimicrobial properties and potential applications in foods – a review. Int. J. Food Microbiol, 94:223-253. DOI: org/10.1016/j.ijfoodmicro.2004.03.022.
12. Koroch A, Juliani HR, Zygadlo JA (2007) Bioactivity of essential oils and their components. Flavours and Fragrances Chemistry, Bioprocessing and sustainability, Berger, R.G., Ed.; Springer Verlag: Berlin, Germany, 87-115. ISBN 978-3-540-49338-9.
13. Delaquis PJ, Stanish K, Girard B, Mazza G (2002) Antimicrobial activity of individual and mixed fractions of dill, cilantro, coriander and eucalyptus essential oils. Int. J. Food Microbiol, 74:101-109. DOI: 10.1016/S0168-1605(01)00734-6.
14. Kotuhov JuA, Anufrieva OA (2016) Artemisia kotuchovii Kupr. (Asteracea) – rare endangered species of Kazakhstan flora. Botanical Research in Siberia and Kazakhstan [Artemisia kotuchovii Kupr. (Asteracea) – redkij ischezajushhij vid flory Kazahstana. Botanicheskie issledovanija Sibiri i Kazahstana] 22:60-70 (In Russian).
15. Red Data Book of Kazakhstan (2014) The 2nd edition revised and supplemented. Vol. 2: Plants. Аstana, LTD «АpтPrintXXI», 222. ISBN 978-601-80334-7-6.
16. Murray P.R., Baron E.J., Pfaller M.A., Tenover F.C., Yolke R.H. Manual of clinical microbiology. 7th ed. – Washington: ASM, 1995. – P. 1773. ISBN 000-1555811264.
17. Kavoosi G, Rowshan V (2013) Chemical composition and antimicrobial activities of essential oil obtained from Ferula assa-foetida oleo-gum-resin: Effect of collection time. Food Chemistry, 138:2180-2187. DOI: 10.1016/j.foodchem.2012.11.131.
18. Schepetkin IA, Kushnarenko SV, Özek G, Kirpotina LN, Utegenova GA, Kotukhov YA, Danilova AN, Özek T, Başer KHC, Quinn MT (2015) Inhibition of human neutrophil responses by essential oil of Artemisia kotuchovii and its constituents. J. Agric. Food Chem, 63:4999-5007. DOI: 10.1021/acs.jafc.5b01307.
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