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Plasmodium knowlesi  is the main cause of malaria in Sarawak, where studies on vectors of  P. knowlesi  have been conducted in only two districts.  Anopheles balabacensis  and  An. donaldi  were incriminated as vectors in Lawas and  An. latens  in Kapit. We studied a third location in Sarawak, Betong, where of 2169 mosquitoes collected over 36 days using human-landing catches, 169 (7.8%) were  Anopheles  spp. PCR and phylogenetic analyses identified  P. knowlesi  and/or  P. cynomolgi, P. fieldi, P. inui ,  P. coatneyi  and possibly novel  Plasmodium spp.  in salivary glands of  An. latens  and  An. introlatus  from the Leucosphyrus Group and in  An. collessi  and  An. roperi  from the Umbrosus Group. Phylogenetic analyses of cytochrome oxidase subunit I sequences indicated three  P. knowlesi -positive  An. introlatus  had been misidentified morphologically as  An. latens,  while  An. collessi  and  An. roperi  could not be delineated using the region sequenced. Almost all vectors from the Leucosphyrus Group were biting after 1800 h but those belonging to the Umbrosus Group were also biting between 0700 and 1100 h. Our study incriminated new vectors of knowlesi malaria in Sarawak and underscores the importance of including entomological studies during the daytime to obtain a comprehensive understanding of the transmission dynamics of malaria.

New vectors that are early feeders for Plasmodium knowlesi and other simian malaria parasites in Sarawak, Malaysian Borneo