The study was carried out to determine relative abundance, species diversity, of Anopheles species (Diptera: Culicidae) in selected forested areas in Cross River State, Nigeria and the prevalence of malaria infection in the specimens. Mosquitoes were collected using pyrethrum spray catch and Centre for Disease Control light traps modified with yeast and sugar to generate carbon dioxide (CO2) and identified using morphological identification keys. We used a multiplex polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) to simultaneously distinguish sibling species of the An. gambiae s.l, including separation of An. gambiae s.s. and An. coluzzii (Diptera: Culicidae).