Malaria vector mosquitoes belong to the genus Anopheles Meigen. The Amazon has been changing due to economic activities such as mining, logging, agriculture, and urbanization, leading to changes in malaria transmission patterns in this region. Therefore, this study aimed to survey Anopheles species in a peri-urban area of Macapá, state of Amapá, Eastern Amazon, Brazil. Human landing catches (HLC) and Shannon light traps were used to collect Anopheles adults at three points. We screened breeding sites in all accessible water reservoirs within a 2 km radius of each collection point to collect immatures and found two species: Anopheles (Nyssorhynchus) albitarsis s.l. Lynch-Arribalzága (35.7%) and Anopheles (Nyssorhynchus) braziliensis (Chagas) (64.3%). A total of 267 winged specimens of Anopheles were collected: A. (N.) braziliensis (62.9%), Anopheles (Anopheles) mattogrossensis Lutz & Neiva (11.6%), Anopheles (Nyssorhynchus) triannulatus s.l. (Neiva & Pinto) (11.6%), A. (N.) albitarsis s.l. Lynch-Arribálzaga (6.7%), Anopheles (Nyssorhynchus) darlingi Root (5.2%), Anopheles (Anopheles) peryassui Dyar & Knab (1.5%), and Anopheles (Nyssorhynchus) nuneztovari Galbadón (0.4%). Considering that Macapá has autochthonous malaria cases, our findings can contribute to developing public health measures in this municipality, therefore helping to protect its habitants.