Background: Studying the ecology of biocontrol-agents is a prerequisite to effectively control medfly (C. capitata) with entomopathogenic fungi. In this context, factors affecting the occurrence and distribution of medfly-associated entomopathogenic-fungi were studied. Soil samples (22) were collected from natural and cultivated areas of Souss-region Morocco. Results: A total of 260 fungal isolates belonging to 22 species and 10 genera were obtained by using medfly pupae as bait. Medfly-associated fungi were detected in all studied soils and pupae infection percentages ranged from 3.33% to 48%. Two genera, Fusarium and Beauveria were the most frequent with 83 isolates (32%) and 50 isolates (19.23%) respectively. Pathogenicity test of isolated species against medfly pupae showed high mortality rates up to 91% for some strains. Principal component analysis (PCA) demonstrated strong influence of origin, physical and chemical properties of soil on the abundance of these fungi. In general, medfly-associated fungi were more abundant in soils with moderate pH (7.5 to 8) having high sand and organic content. High relative humidity negatively influenced the abundance of these fungi. Both factors directly affected the fungal infection percentages in pupae. The response of fungi to these parameters varied among species. According to principal component analysis (PCA) the soils of argan fields and forests were more suitable for the development of medfly-associated fungi than citrus orchards. Conclusions: These results provide guidance on identifying suitable soils for effective application of entomopathogenic fungi as biological control agents. In summary, isolated indigenous strains seem to be a promising option to control C. capitata.