Malaria is a disease transmitted through the bites of female Anopheles infected by Plasmodium sp. Lengkiti, Ogan Komering Ulu (OKU) is a malaria endemic area with a number API of 13,02‰ in 2012. Research to observe the ecological breeding habitat of Anopheles spp. is required as the information in controlling malaria vectors, so that the control can reach right on the target. The aim of this research is to determine the abiotic and biotic factors of Anopheles spp. breeding habitats which can role as vector of malaria in Simpang Empat Village, Lengkiti, OKU, South Sumatra. Larvae collection located in area with stagnant water presumed as potential breeding habitat. The larvae were reared into adult mosquitoes and identified. The found breeding habitats were marshes, ditches, ponds, rubber-soaking ponds, bathing pool, puddle of tire ruts and puddles on road, with characteristics of temperature ranged from 26-38°C, pH ranged from 6-7, with most plants were algae and aquatic Cyperaceae groups; predator of larva were tadpole, Gambusia, Notonecta, Nepa and Hydrometra. Characteristics of abiotic and biotic environment in Simpang Empat Village, Lengkiti, OKU, South Sumatra were suitable as the breeding habitat for the larvae of Anopheles spp.