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). The samples were also screened for Plasmodium infection using the enzyme-linked immunosorbent assay. One hundred and four Anopheles specimens were collected during the study of which 97% was An. gambiae complex and 3% was An. rufipes (Diptera: Culicidae). Only 77% of the An. gambiae s.l. was identify to species level.The result shows that 41.6% was An. gambiae s.s. and 34.6% was An. coluzzii. No sporozoite of Plasmodium was detected in the Anopheles species. The study also found a hybrid form of An. gambiae s.s. and An. coluzzii.These findings suggest the first documented evidence of hybrid forms of An. gambiae s.s./An. coluzzii in South Eastern Nigeria although its epidemiological implication is still not clear.