The research aimed to isolate and characterize local entomopathogenic fungi from different agricultural ecosystems and assess their pathogenicity against cowpea aphid. Forty soil samples were collected from different ecosystems. In total, 38 fungal isolates were collected from the cadavers' exteriors of Galleria mellonella larvae. Microscopic observation of these fungal isolates revealed that they belonged to nine fungal genera among them Metarhizium sp. (3 isolates) and Beauveria sp. (5 isolates). Primary experiments showed different bioactivities of the isolates of the two selected genera against the target insect. Only Metarhizium isolates (M3 & M5) and Beauveria isolates (B1 & B5) were selected for further investigations. Identification of fungal cultures by morphological characters, molecular characterization, ITS sequencing, and NCBI Blast showed that two isolates were confirmed as M. anisopliae and two isolates as B. bassiana. The bioassay evaluation revealed that M. anisopliae (M3 and M5) and B. bassiana (B1) exhibited significant virulence against A. craccivora, as evidenced by their high LT50 and LC50 values and substantial sporulation on the cadavers. Therefore, they could be used for the formulation of pesticides targeting this pest on cowpea and other vegetable crops.