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Background  A small number of human cases of the zoonotic malaria  Plasmodium knowlesi  have been reported in Palawan Island, the Philippines. Identification of potential vector species and their bionomics is crucial for understanding human exposure risk in this setting. Here, we combined longitudinal surveillance with a trap-evaluation study to address knowledge gaps about the ecology and potential for zoonotic spillover of this macaque malaria in Palawan Island.    Methods  The abundance, diversity and biting behavior of human-biting  Anopheles  mosquitoes were assessed through monthly outdoor human landing catches (HLC) in three ecotypes representing different land use (forest edge, forest and agricultural area) across 8 months. Additionally, the host preference and biting activity of potential  Anopheles  vectors were assessed through comparison of their abundance and capture time in traps baited with humans (HLC, human-baited electrocuting net—HEN) or macaques (monkey-baited trap—MBT, monkey-baited electrocuting net—MEN). All female  Anopheles  mosquitoes were tested for the presence of  Plasmodium  parasites by PCR.    Results  Previously incriminated vectors  Anopheles balabacensis  and  An. flavirostris  accounted for > 95% of anophelines caught in longitudinal surveillance. However, human biting densities were relatively low ( An. balabacensis : 0.34–1.20 per night,  An. flavirostris : 0–2 bites per night). Biting densities of  An. balabacensis  were highest in the forest edge, while  An. flavirostris  was most abundant in the agricultural area .  The abundance of  An. balabacensis  and  An. flavirostris  was significantly higher in HLC than in MBT. None of the 357 female  Anopheles  mosquitoes tested for  Plasmodium  infection were positive.    Conclusions  The relatively low density and lack of malaria infection in  Anopheles  mosquitoes sampled here indicates that exposure to  P. knowlesi  in this setting is considerably lower than in neighboring countries (i.e. Malaysia), where it is now the primary cause of malaria in humans. Although anophelines had lower abundance in MBTs than in HLCs,  An. balabacensis  and  An. flavirostris  were caught by both methods, suggesting they could act as bridge vectors between humans and macaques. These species bite primarily outdoors during the early evening, confirming that insecticide-treated nets are unlikely to provide protection against  P. knowlesi  vectors.    Graphical abstractSupplementary Information  The online version contains supplementary material available at 10.1186/s13071-021-04853-9.

The seasonal dynamics and biting behavior of potential Anopheles vectors of Plasmodium knowlesi in Palawan, Philippines