Development of Artificial-Sebum-Containing Leeming and Notman Agar Medium to Enhance the Growth of Malassezia

Modified Leeming and Notman agar medium (mLNA) has been widely utilized to grow lipophilic fungi belonging to the genus Malassezia. We developed a new artificial-sebum-containing mLNA to obtain higher yields of Malassezia species. The olive oil in mLNA was replaced with an artificial sebum composed of triglyceride (triolein), diglyceride (glyceryl distearate), fatty acids (palmitic acid, myristic acid, pentadecanoic acid, and oleic acid), and squalene. Furthermore, the Tween 60 was replaced with self-emulsifying glyceryl stearate. Nine human-associated Malassezia species grew well on the artificial-sebum-containing mLNA, and the most predominant fungus on human skin, Malassezia restricta, exhibited double wet cell weight in artificial sebum-containing mLNA compared to wet cell weight in standard mLNA.