Farklı Origanum spp. (Lamiaceae) uçucu yağlarının kimyasal kompozisyonu ve dört depolanmış ürün zararlısına karşı insektisidal potansiyeli

Öz

Bu çalışmanın amacı dört Origanum türünden elde edilen uçucu yağların kontakt ve fumigant etkinliklerini Rhyzopertha dominica (F., 1792) (Coleoptera: Bostrichidae), Tribolium confusum Jacquelin Du Val, 1863 (Coleoptera: Tenebrionidae), Sitophilus granarius (L., 1875) ve Sitophilus oryzae (L., 1763) (Coleoptera: Curculionidae) erginlerine karşı belirlemektir. Uçucu yağlarının kimyasal kompozisyonu GC-MS cihazı kullanılarak belirlenmiştir. Denemeler laboratuvar koşullarında 2019 yılında Zirai Mücadele Merkez Araştırma Enstitüsü’nde yürütülmüştür. Rhyzopertha dominica ve T. confusum erginlerinde Origanum onites L. ve Origanum vulgare L. var. hirtum uçucu yağları %100 ölüme neden olmuştur. Yirmi dört saat sonunda O. vulgare var. verticium bitki uçucu yağının R. dominica için LD₅₀ değeri 0.046 µl/böcek olarak hesaplanmıştır. Tek konsantrasyon fumigant etki denemeleri sonucunda O. onites ve O. vulgare hirtum uçucu yağları 24 saat sonunda R. dominica’ya karşı yüksek aktivite (sırasıyla %91 ve %70) göstermiştir. Fumigant konsantrasyon etki denemeleri sonucunda bitki uçucu yağlarından O. vulgare uçucu yağı R. dominica için en yüksek etkinliği göstermiş ve 24 saat sonunda LC₅₀ ve LC₉₀ değerleri sırasıyla 0.0052 µl/ml ve 0.0144 µl/ml olarak hesaplanmıştır. Origanum onites’in ana bileşenleri, thymol %22.9; γ-terpinene %13.0; p-cymene %12.9; carvacrol %7.2, O. vulgare var. hitsitum ana bileşenleri carvacrol %32.5; thymol %16.1; p-cymene %12.2; γ-terpinene %7.9, O. vulgare var. verticium ana bileşenleri carvacrol %35.0; p-cymene %11.6; γ-terpinene %10.3; thymol %9.1, O. vulgare x O. onites’in ana bileşenleri carvacrol %15.2; cis-sabinene hydrate %14.6; terpinen-4-ol %14.6 ve γ-terpinene %8.7 olarak belirlenmiştir.

Anahtar Kelimeler

• Kontakt etkinlik,uçucu yağ,fumigant etkinlik,GC-MS,Lamiaceae

Chemical composition and insecticidal potential of different Origanum spp. (Lamiaceae) essential oils against four stored product pests

Abstract

This study was conducted to determine the contact and fumigant toxicity of plant essential oils extracted from four Origanum spp. against four stored product pests, Rhyzopertha dominica (F., 1792) (Coleoptera: Bostrichidae), Tribolium confusum Jacquelin Du Val, 1863 (Coleoptera: Tenebrionidae), Sitophilus granarius (L., 1875) and Sitophilus oryzae (L., 1763) (Coleoptera: Curculionidae). Chemical composition of the essential oils was determined using GC-MS. The trials were conducted under laboratory conditions in 2019 at Plant Protection Central Research Institute. Essential oils extracted from Origanum onites L. and Origanum vulgare L. var. hirtum caused 100% mortality of R. dominica and T. confusum. The computed LD50 value for O. vulgare var. verticium against R. dominica 24 h after application was 0.046 µl/insect. Single concentration fumigant study indicated that O. onites and O. vulgare var. hirtum essential oils cause high mortality (91 and 70%, respectively) of R. dominica within 24 h. Essential oils of O. vulgare showed the highest activity against R. dominica with LC50 and LC90 values of 0.0052 and 0.0144 µl/ml, respectively. The main components of O. onites essential oil were thymol (22.9%), γ-terpinene (13.0%), p-cymene (12.9%) and carvacrol (7.2%). Similarly, the essential oils of O. vulgare var. hirtum were composed of carvacrol (32.5%), thymol (16.1%), p-cymene (12.2%) and γ-terpinene (7.9%). Likewise, the essential oil of O. vulgare var. verticium had carvacrol (35.0%), p-cymene (11.6%), γ-terpinene (10.3%) and thymol (9.1%). Nonetheless, O. vulgare x O. onites essential oil had carvacrol (15.2%), cis-sabinene hydrate (14.6%), terpinen-4-ol (14.6%) and γ-terpinene (8.7%).

Keywords

• Contact activity,essential oils,fumigant activity,GC-MS,Lamiaceae

References

1. Abdelgaleil, S. A., M. I. Mohamed, M. S. Shawir & H. K. Abou-Taleb, 2016. Chemical composition: insecticidal and biochemical effects of essential oils of different plant species from Northern Egypt on the rice weevil. Sitophilus oryzae L. Journal of Pest Science, 89 (1): 219-229.
2. Abou-Taleb, H. K., M. I. Mohamed, M. S. Shawir & S. A. Abdelgaleil, 2016. Insecticidal properties of essential oils against Tribolium castaneum (Herbst) and their inhibitory effects on acetylcholinesterase and adenosine triphosphatases. Natural Product Research, 30 (6): 710-714.
3. Aligiannis, N., E. Kalpoutzakis, S. Mitaku & I. B. Chinou, 2001. Composition and antimicrobial activity of the essential oils of two Origanum species. Journal of Agricultural and Food Chemistry, 49 (9): 4168-4170.
4. Bardakci, H., B. Demirci, E. Yesilada, H. Kirmizibekmez & K. H. C. Başer, 2012. Chemical composition of the essential oil of the subterranean parts of Valeriana alliariifolia. Records of Natural Products, 6 (1): 89-92.
5. Başer, K. H. C., 2009. Uçucu yağlar ve aromaterapi. Fitomed, 7: 8-25.
6. Başer, K. H. C., B. Demirci, F. Demirci, S. Koçak, Ç. Akıncı, H. Malyer & G. Güleryüz, 2002a. Composition and antimicrobial activity of the essential oil of Achillea multifida. Planta Medica, 68 (10): 941-943.
7. Başer, K. H. C., B. Demirci, N. E. Kirimer, F. Satil & G. Tümen, 2002b. The essential oils of Thymus migricus and T. fedtschenkoi var. handelii from Turkey. Flavour and Fragrance Journal, 17 (1): 41-45.
8. Başer, K. H. C., B. Demirci, M. Kurkcuoglu, F. Satin & G. Tumen, 2009. Comparative morphological and phytochemical charactertization of Salvia cadmica and S. smyrnaea. Pakistan Journal of Botany, 41 (4): 1545-1555.

9. Başer, K. H. C., B. Demirci, N. Tabanca, T. Özek & N. Gören, 2001. Composition of the essential oils of Tanacetum armenum (DC.) Schultz Bip., T. balsamita L., T. chiliophyllum (Fisch. & Mey.) Schultz Bip. var. chiliophyllum and T. haradjani (Rech. fil.) Grierson and the enantiomeric distribution of camphor and carvone. Flavour and Fragrance Journal, 16 (3): 195-200.
10. Başer, K. H. C., M. Kürkcüoglu & Z. Aytac, 1998. Composition of the essential oil of Salvia euphratica Montbret ex Bentham var. euphratica from Turkey. Flavour and Fragrance Journal, 13 (1): 63-64.
11. Başer, K. H. C., T. Özek, B. Demirci & H. Duman, 2000. Composition of the essential oil of Glaucosciadium cordifolium (Boiss.) Burtt et Davis from Turkey. Flavour and Fragrance Journal, 15 (1): 45-46.
12. Benelli, G., R. Pavela, R. Petrelli, L. Cappellacci, F. Bartolucci, A. Canale & F. Maggi, 2019. Origanum syriacum subsp. syriacum: From an ingredient of Lebanese ‘manoushe’ to a source of effective and eco-friendly botanical insecticides. Industrial Crops and Products, 134: 26-32.
13. Bond, E. J., 1984. Manual of Fumigation for Insect Control. FAO Plant Production and Protection Paper No: 54, 432 pp.
14. Boukaew, S., P. Prasertsan & S. Sattayasamitsathit, 2017. Evaluation of antifungal activity of essential oils against aflatoxigenic Aspergillus flavus and their allelopathic activity from fumigation to protect maize seeds during storage. Industrial Crops and Products, 97: 558-566.
15. Cardiet, G., B. Fuzeau, C. Barreau & F. Fleurat-Lessard, 2012. Contact and fumigant toxicity of some essential oil constituents against a grain insect pest Sitophilus oryzae and two fungi. Aspergillus westerdijkiae and Fusarium graminearum. Journal of Pest Science, 85 (3): 351-358.
16. Chu, S. S., S. L. Liu, Q. Z. Liu, Z. L. Liu & S. S. Du, 2011. Composition and toxicity of Chinese Dracocephalum moldavica (Labiatae) essential oil against two grain storage insects. Journal of Medicinal Plants Research, 5 (18): 4621-4626.
17. Coccimiglio, J., M. Alipour, Z. H. Jiang, C. Gottardo & Z. Suntres, 2016. Antioxidant. antibacterial. and cytotoxic activities of the ethanolic Origanum vulgare extract and its major constituents. Oxidative Medicine and Cellular Longevity, 2016: 1-8.
18. Conti, B., A. Canale, P. L. Cioni, G. Flamini & A. Rifici, 2011. Hyptis suaveolens and Hyptis spicigera (Lamiaceae) essential oils: qualitative analysis. contact toxicity and repellent activity against Sitophilus granarius (L.) (Coleoptera: Dryophthoridae). Journal of Pest Science, 84 (2): 219-228.
19. Couto, C. S., N. R. Raposo, S. Rozental, L. P. Borba-Santos, L. M. Bezerra, P. A. de Almeida & M. A. Brandão, 2015. Chemical composition and antifungal properties of essential oil of Origanum vulgare Linnaeus (Lamiaceae) against Sporothrix schenckii and Sporothrix brasiliensis. Tropical Journal of Pharmaceutical Research, 14 (7): 1207-1212.
20. da Cunha, J. A., C. de Ávila Scheeren, V. P. Fausto, L. D. W. de Melo, B. Henneman, C. P. Frizzo, R. de Almeida Vaucher, A. C. de Vargas & B. Baldisserotto, 2018. The antibacterial and physiological effects of pure and nanoencapsulated Origanum majorana essential oil on fish infected with Aeromonas hydrophila. Microbial Pathogenesis, 124: 116-121.
21. Demirci, B., K. H. C. Başer & M. Y. Dadandi, 2006. Composition of the essential oils of Phlomis rigida Labill. and P. samia L. Journal of Essential Oil Research, 18 (3): 328-331.
22. Demirci, B., K. H. C. Başer, B. Yıldız & Z. Bahçecioǧlu, 2003. Composition of the essential oils of six endemic Salvia spp. from Turkey. Flavour and Fragrance Journal, 18 (2): 116-121.
23. Demirci, F., D. H. Paper, G. Franz & K. H. C. Başer, 2004. Investigation of the Origanum onites L. essential oil using the Chorioallantoic Membrane (CAM) assay. Journal of Agricultural Food Chemistry, 52 (2): 251-254.
24. Dutra, T. V., J. C. Castro, J. L. Menezes, T. R. Ramos, I. N. do Prado, M. M. Junior, J. M. G. Mikcha & B. A. de Abreu Filho, 2019. Bioactivity of oregano (Origanum vulgare) essential oil against Alicyclobacillus spp. Industrial Crops and Products, 129: 345-349.
25. Erler, F., 2005. Fumigant activity of six monoterpenoids from aromatic plants in Turkey against the two stored-product pests confused flour beetle, Tribolium confusum and Mediterranean flour moth, Ephestia kuehniella. Journal of Plant Diseases and Protection, 112 (6): 602-611.
26. Ertürk, S., A. Yılmaz, T. Akdeniz Fırat & M. Alkan, 2017. Fumigant effect of trans-anethole and carbon dioxide mixture against to Rhyzopertha dominica, Tribolium castaneum and Sitophilus oryzae. Plant Protection Bulletin, 57 (3): 391-400.
27. Giatropoulos, A., A. Kimbaris, A. Michaelakis, D. P. Papachristos, M. G. Polissiou & N. Emmanouel, 2018. Chemical composition and assessment of larvicidal and repellent capacity of 14 Lamiaceae essential oils against Aedes albopictus. Parasitology Research, 117 (6): 1953-1964.
28. Govindarajan, M., S. Kadaikunnan, N. S. Alharbi & G. Benelli, 2016. Acute toxicity and repellent activity of the Origanum scabrum Boiss. & Heldr. (Lamiaceae) essential oil against four mosquito vectors of public health importance and its biosafety on non-target aquatic organisms. Environmental Science and Pollution Research, 23 (22): 23228-23238.
29. Grul'ová, D., M. Pl'uchtová, J. Fejér, L. De Martino, L. Caputo, V. Sedlák & V. De Feo, 2019. Influence of six essential oils on invasive Solidago canadensis L. seed germination. Natural Product Research, 33: 1-3.
30. Guo, S. S., W. J. Zhang, C. X. You, J. Y. Liang, K. Yang, Z. F. Geng & C. F. Wang, 2017. Chemical composition of essential oil extracted from Laggera pterodonta and its bioactivities against two stored product insects. Journal of Food Processing and Preservatio, 41 (2): 1-9.
31. Güncan, A. & E. Durmuşoğlu, 2004. Bitkisel kökenli doğal insektisitler üzerine bir değerlendirme. Hasad Dergisi, 233: 26-32.
32. Hajlaoui, H., H. Mighri, M. Aouni, N. Gharsallah & A. Kadri, 2016. Chemical composition and in vitro evaluation of antioxidant, antimicrobial, cytotoxicity and anti-acetylcholinesterase properties of Tunisian Origanum majorana L. essential oil. Microbial Pathogenesis, 95: 86-94.
33. Ibáñez, M. & M. Blázquez, 2018. Phytotoxicity of essential oils on selected weeds: Potential hazard on food crops. Plants, 7 (4): 1-15.
34. Isman, M. B., 2000. Plant essential oils for pest and disease management. Crop Protection, 19 (8-10): 603-608.
35. Jiang, L. & K. Kubota, 2004. Differences in the volatile components and their odor characteristics of green and ripe fruits and dried pericarp of Japanese pepper (Xanthoxylum piperitum DC.). Journal of Agricultural and Food Chemistry, 52 (13): 4197-4203.
36. Kim, J. & I. K. Park, 2008. Fumigant toxicity of Korean medicinal plant essential oils and components from Asiasarum sieboldi root against Sitophilus oryzae L. Flavour and Fragrance Journal, 23 (2): 79-83.
37. Kim, S. W., H. R. Lee, M. J. Jang, C. S. Jung & I. K. Park, 2016. Fumigant toxicity of Lamiaceae plant essential oils and blends of their constituents against adult rice weevil Sitophilus oryzae. Molecules, 21 (3): 361.
38. Kim, S. I., J. Y. Roh, D. H. Kim, H. S. Lee & Y. J. Ahn, 2003. Insecticidal activities of aromatic plant extracts and essential oils against Sitophilus oryzae and Callosobruchus chinensis. Journal of Stored Products Research, 39 (3): 293-303.
39. Kim, S. I., J. S. Yoon, J. W. Jung, K. B. Hong, Y. J. Ahn & H. W. Kwon, 2010. Toxicity and repellency of origanum essential oil and its components against Tribolium castaneum (Coleoptera: Tenebrionidae) adults. Journal of Asia-Pacific Entomology, 13 (4): 369-373.
40. Kirimer, N., N. Tabanca, T. Özek, G. Tümen & K. H. C. Başer, 2000. Essential oils of annual Sideritis species growing in Turkey. Pharmaceutical Biology, 38 (2): 106-111.
41. Koul, O., S. Walia & G. S. Dhaliwal, 2008. Essential oils as green pesticides: potential and constraints. Biopesticides International, 4 (1): 63-84.
42. Kürkçüoğlu, M., K. H. C. Başer, G. Işcan, H. Malyer & G. Kaynak, 2006. Composition and anticandidal activity of the essential oil of Chaerophyllum byzantinum Boiss. Flavour and Fragrance Journal, 21 (1): 115-117.
43. La Pergola, A., C. Restuccia, E. Napoli, S. Bella, S. Brighina, A. Russo & P. Suma, 2017. Commercial and wild Sicilian Origanum vulgare essential oils: chemical composition, antimicrobial activity and repellent effects. Journal of Essential Oil Research, 29 (6): 451-460.
44. Lee, B. H., P. C. Annis & W. S. Choi, 2004. Fumigant toxicity of essential oils from the Myrtaceae family and 1,8-cineole against 3 major stored-grain insects. Journal of Stored Products Research, 40 (5): 553-564.
45. Lee, B. H., W. S. Choi, S. E. Lee & B. S. Park, 2001. Fumigant toxicity of essential oils and their constituent compounds towards the rice weevil. Sitophilus oryzae (L.). Crop Protection, 20 (4): 317-320.
46. Lesjak, M., N. Simin, D. Orcic, M. Franciskovic, P. Knezevic, I. Beara, V. Aleksic, E. Svircev, K. Buzas & N. Mimica-Dukic, 2016. Binary and tertiary mixtures of Satureja hortensis and Origanum vulgare essential oils as potent antimicrobial agents against Helicobacter pylori. Phytotherapy Research, 30 (3): 476-484.
47. Liang, J. Y., W. T. Wang, Y. F. Zheng, D. Zhang, J. L. Wang, S. S. Guo & J. Zhang, 2017. Bioactivities and chemical constituents of essential oil extracted from Artemisia anethoides against two stored product insects. Journal of Oleo Science, 66 (1): 71-76.
48. López, V., M. Cascella, G. Benelli, F. Maggi & C. Gómez-Rincón, 2018. Green drugs in the fight against Anisakis simplex-larvicidal activity and acetylcholinesterase inhibition of Origanum compactum essential oil. Parasitology Research, 117 (3): 861-867.
49. López, M., M. Jordán & M. Pascual-Villalobos, 2008. Toxic compounds in essential oils of coriander, caraway and basil active against stored rice pest. Journal of Stored Products Research, 44: 273-278.
50. Lourens, A. C. U., D. Reddy, K. H. C. Başer, A. M. Viljoen & S. F. Van Vuuren, 2004. In vitro biological activity and essential oil composition of four indigenous South African Helichrysum species. Journal of Ethnopharmacology, 95 (2): 253-258.
51. Maggio, A., S. Rosselli, M. Bruno, V. Spadaro, F. M. Raimondo & F. Senatore, 2012. Chemical composition of essential oil from Italian populations of Artemisia alba Turra (Asteraceae). Molecules, 17 (9): 10232-10241.
52. Martucci, J. F., L. B. Gende, L. M. Neira & R. A. Ruseckaite, 2015. Oregano and lavender essential oils as antioxidant and antimicrobial additives of biogenic gelatin films. Industrial Crops and Products, 71: 205-213.
53. Mechergui, K., W. Jaouadi, J. P. Coelh & M. L. Khouja, 2016. Effect of harvest year on production. chemical composition and antioxidant activities of essential oil of oregano (Origanum vulgare subsp glandulosum (Desf.) Ietswaart) growing in North Africa. Industrial Crops and Products, 90: 32-37.
54. Mutungi, C., H. Affognon, A. Njoroge, D. Baributsa & L. Murdock, 2014. Storage of mung bean (Vigna radiata [L.] Wilczek) and pigeonpea grains (Cajanus cajan [L.] Millsp) in hermetic triple-layer bags stops losses caused by Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). Journal of Stored Product Research, 58: 39-47.
55. Negehban, M., S. Moharramipour & F. Sefidkon, 2007. Fumigant toxicity of essential oil from Artemisia sieberi Besser against three stored-product insects. Journal of Stored Products Research, 43 (2): 123-128.
56. Özkan, A. M. G., B. Demirci, F. Demirci & K. H. C. Başer, 2008. Composition and antimicrobial activity of essential oil of Ferulago longistylis Boiss. fruits. Journal of Essential Oil Research, 20 (6): 569-573.
57. Polatoğlu, K., B. Demirci, F. Demirci, N. Gören & K. H. C. Başer, 2012c. Biological activity and essential oil composition of two new Tanacetum chiliophyllum (Fisch. & Mey.) Schultz Bip. var. chiliophyllum chemotypes from Turkey. Industrial Crops and Products, 39: 97-105.
58. Polatoğlu, K., F. Demirci, B. Demirci, N. Gören & K. H. C. Başer, 2012a. Essential Oil Composition and Antimicrobial Activities of Tanacetum chiliophyllum Fisch Mey Schultz Bip var monocephalum Grierson from Turkey. Record of Natural Products, 6 (4):184-188.
59. Polatoğlu, K., Ö. C. Karakoç & N. Gören, 2013. Phytotoxic. DPPH scavenging. insecticidal activities and essential oil composition of Achillea vermicularis. A. teretifolia and proposed chemotypes of A. biebersteinii (Asteraceae). Industrial Crops and Products, 51: 35-45.
60. Polatoğlu, K., A. Sen, A. Kandemir & N. Gören, 2012b. Essential oil composition and DPPH scavenging activity of endemic Tanacetum mucroniferum Hub.-Mor. & Grierson from Turkey. Journal of Essential Oil Bearing Plants, 15 (1): 66-74.
61. Polatoğlu, K., H. Servi, Ö. Özçınar, A. Nalbantsoy & S. Gücel, 2017. Essential Oil Composition of Endemic Arabis purpurea Sm. & Arabis cypria Holmboe (Brassicaceae) from Cyprus. Journal of Oleo Science, 66 (1): 65-70.
62. Qari, S. H. & N. A. Abdel-Fattah, 2017. Genotoxic studies of selected plant oil extracts on Rhyzopertha dominica (Coleoptera: Bostrichidae). Journal of Taibah University for Science, 11 (3): 478-486.
63. Reyes-Jurado, F., T. Cervantes-Rincón, H. Bach, A. López-Malo & E. Palou, 2019. Antimicrobial activity of Mexican oregano (Lippia berlandieri). thyme (Thymus vulgaris) and mustard (Brassica nigra) essential oils in gaseous phase. Industrial Crops and Products, 131: 90-95.
64. Shaaya, E., U. Ravid, N. Paster, B. Juven, U. Zisman & V. Pissarev, 1991. Fumigant toxicity of essential oils against four major stored-product insects. Journal of Chemical Ecology, 17 (3): 499-504. Shahriari, M., A. Zibaee, N. Sahebzadeh & L. Shamakhi, 2018. Effects of α-pinene. trans-anethole. and thymol as the essential oil constituents on antioxidant system and acetylcholine esterase of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae). Pesticide Biochemistry and Physiology, 150: 40-47.
65. Shanker, C. & K. R. Solanki, 2000. Botanical insecticides: A historical perspective. India Asian Agrihistory, 4 (2): 21-30.
66. Szczepanik, M., M. Walczak, B. Zawitowska, M. Michalska-Sionkowska, A. Szumny, C. Wawrzeńczyk & M. S. Brzezinska, 2018. Chemical composition, antimicromicrobial activity and insecticidal activity against the lesser mealworm Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) of Origanum vulgare L. ssp. hirtum (Link) and Artemisia dracunculus L. essential oils. Journal of the Science of Food and Agriculture, 98 (2): 767-774.
67. Taban, A., M. J. Saharkhiz & M. Hooshmandi, 2017. Insecticidal and repellent activity of three Satureja species against adult red flour beetles. Tribolium castaneum (Coleoptera: Tenebrionidae). Acta Ecologica Sinica, 37 (3): 201-206.
68. Tabanca, N., B. Demirci, T. Ozek, N. Kirimer, K. H. C. Başer, E. Bedir, I. A. Khan & D.E. Wedge, 2006. Gas chromatographic-mass spectrometric analysis of essential oils from Pimpinella species gathered from Central and Northern Turkey. Journal of Chromatography, 1117 (2): 194-205.
69. Taylor, R. W. D., 1994. Methyl bromide- Is there any future for this noteworthy fumigant? Journal of Stored Products Research, 30: 253-260.
70. Tripathi, A., S. Upadhyay, M. Bhuiyan & P. Bhattacharya, 2009. A review on prospects of essential oils as biopesticide in insect-pest management. Journal of Pharmacogn and Phytotherapy, 1: 52-63.
71. Vinciguerra, V., F. Rojas, V. Tedesco, G. Giusiano & L. Angiolella, 2018. Chemical characterization and antifungal activity of Origanum vulgare, Thymus vulgaris essential oils and carvacrol against Malassezia furfur. Natural Product Research, 4: 1-5.
72. Wijesundara, N. M. & H. V. Rupasinghe, 2018. Essential oils from Origanum vulgare and Salvia officinalis exhibit antibacterial and anti-biofilm activities against Streptococcus pyogenes. Microbial Pathogenesis, 117: 118-127.