Dermatophytosis is a critical sort of skin infection caused by dermatophytes. The long-term treatment of such skin infections may be improved through the application of nanotechnology. This study aimed to prepare griseofulvin zinc Nanohybrid emulsion (GF-Zn–NHE) to improve griseofulvin activity against dermatophytes and some opportunistic pathogenic yeasts and bacteria. The GF-Zn-NHE is prepared by ultra-homogenization ultra-sonication strategies and validated by UV–visible spectroscopy analysis that confirms presences of griseofulvin and Zn-NPs peaks at 265 and 360 nm, respectively. The GF-Zn-NHE has mean distribution size 50 nm and zeta potential in the range from -40 to -36 mV with no significant changes in size distribution and particle size within 120 day ageing. Fourier transform infrared spectroscopy spectrum confirmed the presence of griseofulvin and Zn-NPs stretching vibration peaks. Gamma ray has a negative influence on GF-Zn-NE production and stability. GF-Zn-NHE drug release 95% up to 24 h and 98% up to 72 h of GF was observed and Zinc 90% up to 24 h and 95% up to 72 h, respectively. High antimicrobial activity was observed with GF-Zn–NHE against dermatophytic pathogens in compare with GF, GF-NE, zinc nitrate and ketoconazole with inhibition zone ranged from 14 to 36 mm. The results have shown that the MIC value for Cryptococcus neoformans, Prophyromonas gingivalis and Pseudomonas aeruginosa is 0.125 mg ml ⁻¹ and for Trichophyton rubrum, L. bulgaricus and Escherichia coli value is 0.25 mg ml ⁻¹ and for Candida albicans, Malassezia furfur and Enterococcus faecalis is 0.5 mg ml ⁻¹ and finally 1 mg ml ⁻¹ for Streptococcus mutans. TEM of treated Cryptococcus neoformans cells with GF-Zn-NHE displayed essentially modified morphology, degradation, damage of organelles, vacuoles and other structures.