Confirmation of Anopheles balabacensis as natural vector of malaria caused by Plasmodium knowlesi inhabits forested areas in Kecamatan Balik Bukit, Western Lampung Regency

This study was conducted to confirm the Anopheles spp as malaria vector inhabits forested areas in Western Lampung Regency, Sumatra Island, Indonesia. Anopheles spp. were sampled using the human landing catch method at a remote forest habitat. Each caput and thorax of Anopheles balabacensis specimen was placed separately inside a sterilized mortar and the tissue homogenized using a sterile pestle. The total DNA was extracted from the tissues according to QIAamp DNA Mini Kit protocol. The presence of malaria parasites in the mosquitoes was detected using nested PCR by targeting the small subunit ribosomal RNA (SSU rRNA) gene of Plasmodium. A PCR primer pair, rPLU1, and rPLU5, was used in the first PCR reaction, while another pair (Pkr and Pkf) was used in the second PCR reaction for detecting Plasmodium knowlesi. There was only 1 Anopheles balabacensis mosquito caught in a remote forest habitat during this study. The results of the molecular analysis showed a positive result for Plasmodium knowlesi, which showed a similarity level of 99 with Plasmodium knowlesi isolates from JOHOR02.018S, and Plasmodium knowlesi clone H033H82 isolates from Serawak, and Plasmodium knowlesi genome isolates SWK113.2.18S. It is a confirmed natural vector of Plasmodium knowlesi namely Anopheles balabacensis in a remote forest habitat in Kecamatan Balik Bukit. © 2020 The Authors, published by EDP Sciences.