We assessed the effect of the entomopathogenic fungus Metarhizium anisopliae against Aedes aegypti. Conidia of M. anisopliae strains CG 489, CG 153, and IBCB 481 were grown in Adamek medium under different conditions to improve blastospore production. Mosquito larvae were exposed to blastospores or conidia of the three fungal strains at 1 × 10⁷ propagules mL⁻¹. M. anisopliae IBCB 481 and CG 153 reduced larval survival by 100%, whereas CG 489 decreased survival by about 50%. Blastospores of M. anisopliae IBCB 481 had better results in lowering larval survival. M. anisopliae CG 489 and CG 153 reduced larval survival similarly. For histopathology (HP) and scanning electron microscopy (SEM), larvae were exposed to M. anisopliae CG 153 for 24 h or 48 h. SEM confirmed the presence of fungi in the digestive tract, while HP confirmed that propagules reached the hemocoel via the midgut, damaged the peritrophic matrix, caused rupture and atrophy of the intestinal mucosa, caused cytoplasmic disorganization of the enterocytes, and degraded the brush border. Furthermore, we report for the first time the potential of M. anisopliae IBCB 481 to kill Ae. aegypti larvae and methods to improve the production of blastospores.