Enhancement of Sporulation of Madurella mycetomatis using Natural Substrates and Synthetic Media

Abstract

The fungus Madurella mycetomatis is the common causative agent of mycetoma in Sudan and worldwide. The disease represents a mycological health problem with poor mycological diagnosis. Study of sporulation of M. mycetomatis assists in the identification of the fungus, and helps in the study of its natural habitat and any possible survival structures. In this study, 25 fungal isolates from black grain eumycetoma patients were included. The isolates were specifically identified as M. mycetomatis using species-specific PCR. Media used to enhance the sporulation were: Corn Meal agar, Water agar, Soil agar, Czapek dox agar, and Ascospore agar. In all these media all clinical isolates grew well, while in Ascospore agar only 14 out of the 25 isolates were able to grow. All types of media (except Ascospore agar) supported asexual sporulation of M. mycetomatis, at least in half of the isolates. In Corn Meal agar 13 out of the 25 isolates sporulated, and in Czapek dox agar 12 isolates sporulated. In Water and Soil agar 10 isolates were able to sporulate. v The spores (conidia) were small obovoidal, rounded to pyriform. These conidia were formed on the tip of hyaline well differentiated conidiophores. More than one conidium can be produced from single conidiophore. The mode of conidiogenesis was basipetal non-catenated phialidic enteroblastic conidiogenesis. Some M. mycetomatis isolates were able to produce in vitro grains (sclerotia). Sclerotia were commonly formed on Corn Meal agar. In this medium, 14 out of the 25 isolates produced sclerotia. On Czapek dox agar 7 isolates produced sclerotia, and 3 isolates in Soil agar formed sclerotia, while only 2 isolates formed sclerotia in Water agar. In conclusion, the best medium for sporulation was found to be Corn Meal agar. Czapek dox agar can be used as a medium for growth and sporulation of M. mycetomatis, but Ascospore agar was not a suitable medium for M. mycetomatis growth and sporulation.