As presented at the 104th General Meeting of the American Society for Microbiology, New Orleans, LA, 2004.

Growth Characteristics of Moulds on BBL™ CHROMagar™ Candida Medium
J. L. MORHAIME, T. SCOGNAMIGLIO, S. M. CHUNG, D. H. LARONE Weill Cornell Center, New York-Presbyterian Hospital, New York, NY

ABSTRACT
■ BACKGROUND: BBL CHROMagar Candida (CAC, Becton Dickinson, Sparks, MD) is a selective medium that is primarily used for the isolation of yeasts and the rapid identification of some Candida species based on differential pigment production. Although the manufacturer states that filamentous fungi can also be isolated, there have been no studies as to the efficiency of CAC in supporting their growth nor documentation of their resultant macroscopic and microscopic morphology. This study sought to determine whether clinically encountered moulds can indeed grow on CAC and, if so, to ascertain their macroscopic and microscopic appearance. METHODS: Stock cultures of 20 moulds recovered from clinical specimens were revitalized on Sabouraud Dextrose Agar (SDA) and subsequently inoculated onto CAC and incubated at 30ºC. The isolates were representative of the following fungal groups: thermally dimorphic, zygomycete, dematiaceous, dermatophyte, and hyaline hyphomycete. Rate of growth, colony texture, and color of colony surface and reverse were recorded. Upon maturity, microscopic morphology was examined in wet mount preparations made with lactophenol cotton blue and PVA. RESULTS: All study organisms grew satisfactorily on CAC with an initial green color seen on the reverse of the colonies. When mature, the mould colonies exhibited their classic texture. Surface pigmentation was sometimes less than that generally seen on SDA, and the typical reverse pigment was often replaced by a green hue. Wet preps of the colonies demonstrated typical microscopic morphology for each of the organisms studied. CONCLUSION: Clinically encountered moulds are able to grow on CAC and often produce colonies that are similar to those seen on more conventional agar, e.g., SDA. For those that are divergent from their classic pigmentation, their typical microscopic morphology can at least suggest an identification. A slant or plate and slide culture on routine mycology agar can be performed if required. When CAC is used as the sole medium for specimens from which only yeasts are usually anticipated, unsuspected moulds will also be recovered if present. INTRODUCTION • CHROMagar Candida (CAC, Becton Dickinson, Sparks, MD) - A selective medium primarily used for the isolation of yeasts - Rapid identification of some Candida species based on differential pigment production - Inhibition of bacterial growth • Manufacturer states that filamentous fungi can also be isolated - No studies have been done to show the efficiency of CAC in supporting their growth - No documentation of their resultant macroscopic and microscopic morphology • Questions to answer in this study: - Can clinically encountered molds grow on CAC? - What is the resultant macroscopic and microscopic appearance?

MATERIALS AND METHODS • Stock cultures of 20 moulds recovered from clinical specimens were revitalized on Sabouraud Dextrose Agar (SDA). Five fungal groups were represented: - Thermally Dimorphic Fungi (1): Histoplasma capsulatum - Zygomycetes (2): Rhizopus, Absidia - Dematiaceous Fungi (6): Cladosporium sp., Pseudallescheria boydii, Wangiella, Alternaria, Pithomyces, Chaetomium - Dermatophytes (3): Microsporum canis, Trichophyton rubrum, Trichophyton tonsurans - Hyaline Hyphomycetes (8): Emmonsia, Aspergillus spp (fumigatus, niger, flavus, terreus), Penicillium, Paecilomyces, Fusarium • Each isolate was subsequently inoculated onto CAC and incubated at 30ºC • Rate of growth, colony texture, color of colony surface and reverse was recorded • Upon maturity, microscopic morphology was examined in wet mount preparations made with lactophenol cotton blue and PVA

g. and Microscopic Morphology of Representative Organisms from Five Fungal Categories Grown on BBL CHROMagar Candida Agar Histoplasma capsulatum Microsporum canis Aspergillus fumigatus Rhizopus Alternaria Microscopic Morphology Reverse Surface RESULTS • All study organisms grew satisfactorily on CAC • Upon maturation the mould colonies exhibited their classic texture • Surface pigmentation was sometimes less than that generally seen on SDA • In many instances. SDA • For those that differ from their classic pigmentation. the reverse of the colonies showed a green color • Wet preps of the colonies demonstrated typical microscopic morphology for each of the organisms studied DISCUSSION • In our study. the mould isolates often produced colonies that are similar to those seen on more conventional agar. if present • Moulds grown on CAC may or may not have typical pigment and/or macroscopic morphology • Microscopic morphology is consistently typical of the organism on CAC LR810 . the clinically encountered moulds are also able to grow.Characteristic Surface. microscopic morphology can suggest an identification. e.. Reverse. A slant (or plate) and slide culture on routine mycology agar can be performed if required CONCLUSION • When CHROMagar Candida Agar is used for initial isolation of yeasts from specimens for fungal culture.