An entomological study was conducted in a village of Sudano-Guinean savanna in Senegal, during the rainy season from July to November 2001, to investigate the biology and the involvement of each anopheline species in malaria transmission. Mosquitoes were captured when landing on human volunteers and by pyrethrum spray catches. Twelve anopheline species were captured. Four species amounted to 97% of human-bait sampling: Anopheles gambiae molecular form S, An. arabiensis, An. funestus, and An. nili s.s. All An. gambiae and An. nili females were fed on human, whereas the anthropophilic rate was 94.5% for An. funestus and 88.9% for An. arabiensis. Plasmodium falciparum was the only malaria parasite found, and infecting only An. gambiae, An. arabiensis, An. funestus, and An. nili. The circumsporozoite rate was 4.5% for An. gambiae, 1.6% for An. arabiensis, 3.9% for An. funestus, and 2.1% for An. nili. During the period of study, the entomological inoculation rate was estimated to 264 infected bites. An. gambiae, An. arabiensis, An. funestus, and An. nili were responsible respectively of 56, 3, 20, and 21% of malaria transmission. This study shows for the first time the implication of An. nili in malaria transmission in this area and the complexity of the malaria vectorial system that should be taken into account for any malaria control strategy.