FLORA OF SCALP AND DANDRUFF 11• Glucose ........................... 20.0 MgSO4' 7H•O ...................... 0. ,5 KH2PO4 .......................... 1.0 Novobiocin* ........................ 0.1 Griseofulvint ....................... 0. 025 To each liter was added 10 ml of a vitamin concentrate which con- tained in micrograms: Biotin ............................ 2.0 Thiamine hydrochloride ............ 400.0 Inositol ............................ 2000.0 The glucose salts and novobiocin were autoclaved in 980 ml of dis- tilled water. Griseofulvin (250 mg) was autoclaved in the dry form in a 100-ml volumetric flask. Sufficient water was added to make a volume of 100 and 10 ml was pipetted into the medium. The vitamin concen- trate was sterilized by filtration and was refrigerated until ready for use. Basal Medium II for Molds--Modification of Czapek Dox Broth (Difco). It contained in grams per liter of distilled water: Saccharose ........................ 30.0 K2HPO4 .......................... 1.0 MgSO4' 7H,,,O ...................... 0.5 KC1 ............................... 0.5 FeSO4 ............................ 0.01 Novobiocin ........................ 0.1 The ingredients were dissolved and the solution was autoclaved. Basal Medium III.for Bacteria--Modification of Czapek Dox Broth (Difco). The sugar and salts in this medium were the same as those in Medium II. The antibiotics used were 100 units of nystatin1: per ml and 0.025 g of griseofulvin per liter. The nystatin was added to the medium with a sterile syringe. * Novobio½in powder (Albamycin, sodium Lot. No. Sm-577) was supplied by the Upjohn Company, Kalamazoo, Mich. t Griseofulvin powder (Grifulvin, Lot No. 2214) was supplied by McNeil Laboratories, Fort Washington, Pa. • Nystatin (Mycostatin) was supplied by E. R. Squibb and Sons, New York, N.Y. It came in a sterile vial with 500,000 units of powder (List 5918).

120 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS Basal Medium IV for Actinomycetes--Modification of Noval and Nickerson (21). It contained in grams per liter of distilled water: Glucose ........................... 2.0 K2HPO4 .......................... 1.5 MgSO4' 7H20 ...................... 0.05 CaC12 ............................. 0.05 FeSO4.7H20 ....................... 0. 015 ZnSO4- 7H20 ....................... 0. 005 The ingredients were dissolved and the solution was autoclaved. This medium contained nystatin (100 units per ml) which was added with a sterile syringe. Basal Medium V for Yeast and Molds--This medium contained in grams per liter of distilled water: Novobiocin ........................ 0.1 K2HPO• .......................... 1.0 The ingredients were dissolved and the solution was autoclaved. The novobiocin was added to minimize bacterial contamination and the K2HPO• served as a buffer. The only nutrients available as carbon and nitrogen sources were the natural constituents of scurf. Basal Medium VI for Bacteria and Actinomycetes--This medium is similar to Medium V in that it has no added nutrients. Each liter of distilled water contained: K2HPO4 .......................... 1.0 g Nystatin ................. 100,000.0 units Techniques for Isolation of Resident Flora Four isolation techniques were carried out initially on six subjects to determine the best method. These methods included the use of Noble agar base cultures, silica gel base cultures, flask broth cultures, and per- colation apparatus cultures. Because the last isolation procedure pre- vented any variation in incubation temperature, an incubation tempera- ture of 37 øC was chosen for all four methods. All isolation cultures were incubated for one month or until over- grown or desiccated before discarding. The four procedures were as follows: Noble Agar Base Cultures*--A 1.5% concentration of Special-Agar * The isolates from all four methods were similar, so that the use of Noble Agar Base be- came the method of choice for the remaining 46 subjects.