The study evaluates the resistance and susceptibility of adult female Anopheles gambiae s. l., Aedes aegypti and Culex quinquefasciatus mosquitoes sourced within Osogbo metropolis, Osun State, Nigeria to four groups of insecticides [Permethrin, Deltamethrin, Pirimiphos-methyl and DDT (Dichlorodiphenyltrichloroethan)] and the distribution of their larval habitat within the metropolis. Mosquito larvae of the three genera were collected during the wet season and reared to adult stage in the laboratory. Emerged adult female mosquitoes were exposed to insecticide impregnated papers of the four insecticide groups for 60mins using WHO kits to determine the knock down rate (kdr). Thereafter, they were to holding tubes and left for 24hrs to assess their resistance and susceptibility according to the WHO protocol. Four types of larval habitats were identified (tires, ground pools, gutters and plastic containers). Anopheles gambiae s. l. showed the greatest resistance to Permethrin (49%) (p=0.04, p<0.05) while the highest susceptibility was recorded with Pirimiphos-methyl (69%) with the lowest against Permethrin (16%) (P=0.002; p<0.05). The greatest resistance of A. aegypti was against OC-Control (45%) (p=0.031; p<0.05). Permethrin had the highest susceptibility (60%) against A. aegypti while OC-control had the lowest (11%) (p= 0.005; p< 0.05). Culex quinquefasciatus had a lesser resistance to OC-control (38%) as compared with Aedes aegypti (45%). However, it was most least susceptible to Pirimiphos-methyl (52%) and DDT (17%) respectively (p=0.013; p<0.05). The susceptibility of A. gambiae s. l. and C. quinquefasciatus to Pirimiphos-methyl and A. aegypti to Permethrin is an indication of the possibility of success if employed for vector control of A. gambiae s. l., C. quinquefasciatus and A. aegypti respectively. This could be through their inclusion as active ingredients in insecticide treated nets (ITNs) and indoor residual spray (IRS) with a view to abating malaria and other life-threatening mosquito-borne diseases constituting global public health scourge.