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Background  Environmental disturbance, deforestation and socioeconomic factors all affect malaria incidence in tropical and subtropical endemic areas. Deforestation is the major driver of habitat loss and fragmentation, which frequently leads to shifts in the composition, abundance and spatial distribution of vector species. The goals of the present study were to: (i) identify anophelines found naturally infected with  Plasmodium ; (ii) measure the effects of landscape on the number of  Nyssorhynchus darlingi , presence of  Plasmodium -infected Anophelinae, human biting rate (HBR) and malaria cases; and (iii) determine the frequency and peak biting time of  Plasmodium -infected mosquitoes and  Ny. darlingi.Methods  Anopheline mosquitoes were collected in peridomestic and forest edge habitats in seven municipalities in four Amazon Brazilian states. Females were identified to species and tested for  Plasmodium  by real-time PCR. Negative binomial regression was used to measure any association between deforestation and number of  Ny. darlingi , number of  Plasmodium -infected Anophelinae, HBR and malaria. Peak biting time of  Ny. darlingi  and  Plasmodium -infected Anophelinae were determined in the 12-h collections. Binomial logistic regression measured the association between presence of  Plasmodium -infected Anophelinae and landscape metrics and malaria cases.    Results  Ninety-one females of  Ny. darlingi ,  Ny. rangeli ,  Ny. benarrochi  B and  Ny. konderi  B were found to be infected with  Plasmodium.  Analysis showed that the number of malaria cases and the number of  Plasmodium -infected Anophelinae were more prevalent in sites with higher edge density and intermediate forest cover (30–70%). The distance of the drainage network to a dwelling was inversely correlated to malaria risk. The peak biting time of  Plasmodium -infected Anophelinae was 00:00–03:00 h. The presence of  Plasmodium -infected mosquitoes was higher in landscapes with > 13 malaria cases.    Conclusions   Nyssorhynchus darlingi ,  Ny. rangeli ,  Ny. benarrochi  B and  Ny. konderi  B can be involved in malaria transmission in rural settlements. The highest fraction of  Plasmodium -infected Anophelinae was caught from midnight to 03:00 h. In some Amazonian localities, the highest exposure to infectious bites occurs when residents are sleeping, but transmission can occur throughout the night. Forest fragmentation favors increases in both malaria and the occurrence of  Plasmodium -infected mosquitoes in peridomestic habitat. The use of insecticide-impregnated mosquito nets can decrease human exposure to infectious Anophelinae and malaria transmission.Supplementary Information  The online version contains supplementary material available at 10.1186/s13071-021-04725-2.

Vector role and human biting activity of Anophelinae mosquitoes in different landscapes in the Brazilian Amazon