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Both Gossypium barbadense (GB) and Gossypium hirsitum (GH) are members of the Malvaceae family and the cotton genus. In order to reduce silver nitrate, aqueous extracts of both plants were used to synthesize biogenic silver nanoparticles (AgNPs) (AgNO3). The techniques employed to describe them included X-ray diffraction, ultraviolet-visible spectroscopy, scanning electron microscopy, energy dispersive X-ray (EDX), transmission electron microscopy (TEM), and the fourier transformed infrared (FTIR) spectrophotometer. The AgNPs were found to be crystalline according to the XRD spectra, but TEM images showed that they were evenly distributed, free of aggregation, and in irregular shapes with an average size of 21 nm. Only silver, oxygen, and carbon were present in the nanoparticles, according to the SEM and EDX data. Both AgNPs have excellent antimalarial efficacy when tested in vitro with Plasmodium falciparum, with IC50 values of 1.2 and 0.96 g/mL, weak antitrypanosomal potentials, and a good track record of negligible cytotoxicity against the HeLa cell line. Both AgNPs exhibit potent antiplasmodial and antibacterial characteristics, making them intriguing candidates for use in nanomedicines and other contexts where related applications are needed. The aim of this study is to biogenically synthesize silver nanoparticles (AgNPs) using the aqueous leaf extracts of GB and GH, analyze their ability to target the HeLa cell line, and determine whether they have antimalarial and antitrypanosomal properties. [ABSTRACT FROM AUTHOR]