Keratinous substance rich soil is most conducive for keratinophilic fungi growth and occurrence. Dermatophytes and other related fungi are potential pathogens causing human and animal dermatomycoses. Herein, this study went for screening the presence of dermatophytes and related keratinophilic fungi in various soil samples collected from different locations of Sharkia Governorate, Egypt. A total of 80 soil samples from roadsides, fields and stables were subjected for mycological analysis using modified hair-bait technique with hair of horse, cattle and goat, sheep wool, and chicken feathers as a keratin source for keratinophilic fungi growth. Keratinophilic fungi were identified according to their phenotypical characterization in combination with PCR amplification and sequencing for internal transcribed spacer (ITS) region of rDNA. Keratinophilic fungi were recovered from 73.75% of soil samples (59/80). Field soils yielded a higher positivity rate for keratinophilic fungal isolates (90%) than roadsides (66.67%) and stables (58.82%). The majority of keratinophilic fungi belonged to dermatophytes (57.47%). Microsporum gypseum (50.85%) were detected in the majority of sites followed by Trichophyton mentagrophytes (30.51%), Chrysosporium species (28.81%), C. keratinophilium (23.73%), C. tropicum, C. zonatum, Arthroderma multifidum, Arthroderma benhami, Arthroderma fulvum, Clonostachys species, Simplicillium obclavatum and Purpureocillium lilacinum (1.69%, each). It was found that horse and goat hair were more suitable for isolation of keratinophilic fungi with a percentage of 100% for each, followed by cattle hair (91.66%), sheep wool (87.5%) and chicken feathers (83.33%). This investigation demonstrated that the various soils of Sharkia Governorate might be critical suppliers of certain keratinophilic fungi that may constitute hazards to human and animal health. The genetic-based identification is strongly recommended for a high discriminatory identification of keratinophilic fungi.