The Efficacy of Piper betle L. (betel vine) Essential Oil against Malassezia pachydermatis Isolated from Skin Lesions of Canine Dermatitis

Main Article Content

Jareerat Aiemsaard
Pittaya Papirom
Setthat Likitchatpakdee
Eakachai Thongkham

Abstract

Resistance to antifungal drugs is a very important problem in the treatment of yeast infections in dogs. Therefore, it is important to study new substances to promote the treatment of Malassezia dermatitis in dogs. The aim of this study was to evaluate the antifungal activity of betel vine essential oil against Malassezia pachydermatis isolated from skin lesions of canine dermatitis by broth microdilution method and time kill test. The essential oil was analyzed for its chemical properties by gas chromatography/mass spectrometry (GC/MS). The GC/MS analysis revealed that eugenol was the major component of the tested betel vine essential oil (32.82%) followed by eugenol acetate (26.51%), 4-allyl-1,2-diacetoxybenzene (15.85%) and p-chavicol (7.45%). The results of antifungal testing demonstrated that betel vine essential oil had high antifungal activity against all tested M. patchydermatis isolates, with minimum inhibition concentration (MIC) and minimum fungicidal concentration (MFC) values equal to 0.66 and 0.66-1.13 µg/ml, respectively. The time-kill assay showed that a concentration of 8 times the MIC had the highest activity followed by 4 times the MIC, which eradicated tested microorganisms within 30 minutes by more than 99.9% and 90%, respectively. These activities appear to be dependent on dose and time and might be affected by the chemical constituents. These results revealed that betel vine essential oil has the potential to be developed as a drug for use against M. pachydermatis. Further studies to determine the suitable concentration and formulation, and for in vivo antifungal efficacy are required.

Article Details

Section
Research Article

References

Adorjan, B., and G. Buchbauer. 2010. Biological properties of essential oils: an updated review. Flavour. Fragr. J. 25(6): 407-426.

Aiemsaard, J., and K. Punareewattana. 2017. Antifungal activities of essential oils of Syzygium aromaticum, Piper betle, and Ocimum sanctum against clinical isolates of canine dermatophytes. ScienceAsia. 43(4): 223-228.

Aiemsaard, J., S. Aiumlamai, S. Taweechaisupapong, C. Aromdee, and W. Khunkitti. 2010. Chemical composition, antioxidant activity and antibacterial action of eight essential oils against clinical isolates of mastitis pathogens. Int. J. Essent. Oil. Ther. 4: 37-43.

Ali, B., N. A. Al-Wabel, S. Shams, A. Ahamad, S. A. Khan, and F. Anwar. 2015. Essential oils used in aromatherapy: A systemic review. Asian. Pac. J. Trop. Biomed. 5(8): 601-611.

Ali, I., F. G. Khan, K. A. Suri, B. D. Gupta, N. K. Satti, P. Dutt, et al. 2010. In vitro antifungal activity of hydroxychavicol isolated from Piper betle L. Ann. Clin. Microbiol. Antimicrob. 9: 7.

Ali, I., N. K. Satti, P. Dutt, and R. Prasad. 2016. Hydroxychavicol: A phytochemical targeting cutaneous fungal infections. Sci. Rep. 6: 37867.

Amelia, B., E. Saepudin, A. H. Cahyana, D. U. Rahayu, A. S. Sulistyoningrum, and J. Haib. 2017. GC-MS analysis of clove (Syzygium aromaticum) bud essential oil from Java and Manado. AIP. Conf. Proc. 1862: 030082.

Asawapattanakul, T, W. Khunkitti, R. Leesing, and J. Aiemsaard. 2013. The efficacy of essential oils against Malassezia pachydermatis isolated from canine otitis externa. Isan. J. Pharm. Sci. 9: 40-51.

Cafarchia, C., S. Gallo, G. Capelli, and D. Otranto. 2005. Occurrence and population size of Malassezia spp. in the external ear canal of dogs and cats both healthy and with otitis. Mycopathologia. 160(2): 143-149.

Crespo, M. J., M. L. Abarca, and F. J. Cabanes. 2002. Occurrence of Malassezia spp. in the external ear canals of dogs and cats with and without otitis externa. Med. Mycol. 40(2): 115-121.

da Silva Nascente, P., A. R. M. Meinerz, R. O. de Faria, L. F. D. Schuch, M. C. A. Meireles, and J. R. B. de Mello. 2009. CLSI broth microdilution method for testing susceptibility of Malassezia pachydermatis to thiabendazole. Brazilian. J. Microbiol. 40: 222-226.

Dagli, N., R. Dagli, R. S. Mahmoud, and K. Baroudi. 2015. Essential oils, their therapeutic properties, and implication in dentistry: A review. J. Int. Soc. Prevent. Communit. Dent. 5(5): 335-340.

Department of medical sciences. 2014. Standard Method for Food Analysis Volume 2; DMSc F 2009 Analytical method of yeasts and molds in food by pour plate and spread plate. National Office of Buddhism publishing. Bangkok. 211 p.

Ernst, E. J., D. J. Keele, M. E. Klepser, C. R. Petzold, and E. E. Roling. 2002. In vitro activity of micafungin (FK-463) against Candida spp.: microdilution, time-Kill, and post antifungal-effect studies. Antimicrob. Agents. Chemother. 46(12): 3846-3853.

Fan, Y. M., W. M. Huang, S. F. Li, G. F. Wu, K. Lai, and R. Y. Chen. 2006. Granulomatous skin infection caused by Malassezia pachydermatis in a dog owner. Arch. Dermatol. 142(9): 1181-1184.

Kaypetch, R., and S. Thaweboon. 2018. Antifungal property of Piper betle leaf oil against oral Candida species. MATEC. Web. Conf. 242: 01021.

Kumar, A., K. Singh, and A. Sharma. 2002. Prevalence of Malassezia pachydermatis and other organisms in healthy and infected dogs ears. Isr. Vet. Med. Assoc. 57(4): 145–148.

Mamilla, R., C. Raja, V. Srinivasan, S. Selvaraj, and S. K. Mahapatra. 2015. Versatile and synergistic potential of eugenol: A review. Pharmaceutica. Analytica. Acta. 6(5): 1000367.

Markey, B. K., F. C. Leonard, M. Archambault, A. Cullinane, and D. Maguire, editor. 2013. Clinical veterinary microbiology. 2nd ed. Elsevier, Missouri.

Mohottalage, S., R. Tabacchi, and P. M. Guerin. 2007. Components from Sri Lankan Piper betle L. leaf oil and their analogues showing toxicity against the housefly, Musca domestica. Flavour Fragr. J. 22(2): 130-138.

Nakano, Y., M. Wada, H. Tani, K. Sasai, and E. Baba. 2005. Effects of beta-thujaplicin on anti-Malassezia pachydermatis remedy for canine otitis externa. J. Vet. Med. Sci. 67(12): 1243-1247.

Nguyen, T. N. T., T. N. N. Huynh, V. T. Tran, C. H. Dang, T. K. D. Hoang, and T. D. Nguyen. 2018. Physicochemical characterization and bioactivity evaluation of essential oils from Citrus microcarpa Bunge leaf and flower. J. Essent. Oil. Res. 30(4): 285-292.

Nijima, M., R. Kano, M. Nagata, A. Hasegawa, and H. Kamata. 2011. An azole-resistant isolate of Malassezia pachydermatis. Vet. Microbiol. 149(1-2): 288-290.

Park, M. J., K. S. Gwak, I. Yang, W. S. Choi, H. J. Jo, J. W. Chang, et al. 2007. Antifungal activities of the essential oils in Syzygium aromaticum (L.) Merr. Et Perry and Leptospermum petersonii Bailey and their constituents against various dermatophytes. J. Microbiol. 45: 460-465.

Ravindran, P. N., and G. S. Pillai. 2004. Under-utilized herbs and spices. In: K.V. Peter, editor, Handbook of Herbs and Spices Volume 2, Woodhead Publishing, Cambridge, pp 52-607. 68-102.

Row, L. M., and J. Ho. 2009. The antimicrobial activity, mosquito larvicidal activity, antioxidant property and tyrosinase inhibition of Piper betle. J. Chin. Chem. Soc. 56(3): 653-658.

Sugumaran, M., M. Suresh Gandhi, K. Sankarnarayanan, M. Yokesh, M. Poornima, and R. Sreerama. 2011. Chemical composition and antimicrobial activity of vellaikodi variety of Piper betle Linn leaf oil against dental pathogens. Int. J. PharmTech. Res. 3(4): 2135-2139.

Thosar, N., S. Basak, R. N. Bahadure, and M. Rajurkar. 2013. Antimicrobial efficacy of five essential oils against oral pathogens: An in vitro study. Eur. J. Dent. 7(1): S071-S077.

Usman, M. S., M. E. E. Zowalaty, K. Shameli, N. Zainuddin, M. Salama, and N. A. Ibrahim. 2013. Synthesis, characterization, and antimicrobial properties of copper nanoparticles. Int. J. Nanomed. 8: 4467-4479.

Wiebe, V. J., editor. 2015. Drug therapy for infectious diseases of the dog and cat. John Wiley & Sons, Iowa.

Yurayart, C., A. Chindemporn, S. Suradhat, P. Tummaruk, S. Kajiwara, and N. Prapasarakul. 2011. Comparative analysis of the frequency, distribution and population sizes of yeasts associated with canine seborrheic dermatitis and healthy skin. Vet. Microbiol. 148(2-4): 356-362.