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.

Separation of viable histamine-producing bacteria from yellowtail meat components by density gradient centrifugation.

Quantitative separation of live cells from food samples is essential for non-culture methods to be validated. In this viewpoint, the feasibility of density gradient centrifugation (DGC) was demonstrated for the first time using samples of yellowtail meat to which Morganella morganii, a histamine producing bacterium had been added. Using a Ficoll density gradient from 50 to 10 w/v % with 10 w/v % steps, meat-free fractions of M. morganii cells were collected in 20-50 w/v % layers. The total cell collection rate ranged from 73-86 % irrespective of the cell density in the range 10(2)-10(6) cells/200 microl.