RESEARCH ARTICLE New Medium for Rapid Diagnosis and Determination of Antifungal Testing Against Malassezia spp.: A Potential Candidate for Industries Anupam Dikshit • Amit Kumar Tiwari • Rohit Kumar Mishra Received: 24 May 2012 / Revised: 5 September 2012 / Accepted: 17 October 2012 / Published online: 13 February 2013 Ó The National Academy of Sciences, India 2013 Abstract During in vitro investigation of herbal antimi- crobials against Malassezia sp., the causal organism of human skin diseases, such as pityriasis versicolor, seborrhoeic dermatitis, dandruff etc., scientists have faced several problems due to improper media. To overcome these problems, the present invention provided a selective broth medium, for excellent growth of all known Malas- sezia sp. in very short duration of 2–3 days, having least chances of contamination, easy method of preparation. Besides, the new medium was also found useful for cul- turing human dermatophytes and other unicellular yeast. In addition, the use of the modified medium will provide rapid diagnosis as well as determination of antifungal evaluation in vitro through CLSI method. As such, the medium pos- sess enormous potential for industrial production. The invention has been filed for patent on February 27, 2012 (application no. DEL/546/2012) through Patent Facilitating Cell, Technology Information, Forecasting and Assessment Council (TIFAC) of the Department of Science and Technology (DST) at Indian Patent Office, New Delhi. Keywords Media applied for patent  Antimicrobial testing  Dandruff  Malassezia  Pityriasis versicolor Introduction Recent advances in novel and multidisplinary research along with budding technologies have contributed to a great extent in explaining that various species of the genus Malassezia played noteworthy role in skin diseases. Currently 14 species of Malassezia are known, multidisciplinary research is likely to show the etiopathological relationships between new species and pityriasis versicolor, seborrhoeic dermatitis, dandruff etc. [1, 2]. Discovery of new methods is still needed for the prevention, treatment and management of infections caused by Malassezia spp. Advancement of new antifungal compounds has increased the demand for in vitro testing method [3]. Clinical and Laboratory Standards Institute (CLSI) have recommended broth microdilution methods to determine antimicrobial susceptibility testing of yeast using 96 well plate in year 2002, 2003 and their methods are globally acceptable [4, 5]. CLSI has recommended RPMI 1640 medium which is not suitable for entire range of Malassezia sp.. There is still no single medium that is suitable for growth and maintenance of all the species of Malassezia described by Ashbee [6]. As such, forms the basis for further modification in the patented medium titled ‘‘Modified Culture Medium and process for the preparation thereof’’ applied for patent in 2002 and granted in 2008, (Dutta and Dikshit [7], patent no. 217897) through Patent Facilitating Cell, Tech- nology Information, Forecasting and Assessment Council (TIFAC) of the Department of Science and Technology (DST) at Indian Patent Office, New Delhi. Although this medium was found good for sub-culturing of few Malassezia sp. and traditional antifungal susceptibility testing like disc diffusion, but the patented medium was found unfit for broth micro dilution method. To solve the aforementioned problem, present novel invention is carried out and succeeded in developing new medium for enhanced growth for number of A. Dikshit (&)  A. K. Tiwari  R. K. Mishra Biological Product Laboratory, Department of Botany, University of Allahabad, Allahabad 211002, India e-mail: anupambplau@rediffmail.com A. K. Tiwari e-mail: amitstar82@gmail.com R. K. Mishra e-mail: rohit_ernet@yahoo.co.in 123 Natl. Acad. Sci. Lett. (January–February 2013) 36(1):61–66 DOI 10.1007/s40009-012-0081-3