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김주연,Thierry Olivry,손원근 한국임상수의학회 2012 한국임상수의학회지 Vol.29 No.2
This work describes the characteristics of Malassezia pachydermatis isolated from dog ear canals and the effect of essential oils on the growth of this organism. Sterile cotton swabs were used to collect specimens from the external ear canal and culture tests were performed to detect the population size of Malassezia yeast. Using three different isolation media, included Sabouraud dextrose agar (SDA) to isolate common M. pachydermatis, and SDA supplemented with olive oil (SDAO) and Leeming's medium (LM) to detect lipophilic yeast, Malassezia spp were isolated from 14 of 18 dogs (77.8%); isolation rates were 33.3% in SDA, 72.2% in SDAO and 66.7% in LM media. All Malassezia spp isolates were identified as M. pachydermatis according to results of PCR amplification, but gross colony morphology and SDA growth rates suggested four different subtypes. Large (LC) and medium colony (MC)types respectively describe large colony (diameter > 3 mm) and medium colony (around 2 mm) after 72 hour incubation,and small (SC) type refers to smaller colony (< 1 mm) even after 5 days incubation; lipid dependent colonies did not grow onto SDA. Large Colony type strains were isolated from 4, 11, and 11 samples, MC type strains from 2,3 and 1 and SC type strains from 1, 2 and 1 in SDA, SDAO and LM, respectively. Lipid-dependent M. pachydermatis (Lipo) were isolated from 3 samples each in SDAO and LM. Anti-M. pachydermatis activity testing was done using disc-diffusion assays and well diffusion tests. Most essential oils inhibited the growth of M. pachydermatis in a range from 0.5% to 1.0% of essential oils. MIC90 and MIC50 were variable depending upon the nature of essential oils. Thyme oil was found to be highly effective in inhibiting the growth of M. pachydermatis in a range from 0.125%to 0.0625% while marjoram and then tea tree oil exhibited lower inhibitory capacity.