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Tracking Plasmodium knowlesi through faecal DNA for monitoring zoonotic transmission in wild macaques across Southeast and South Asia

Wannigama, Dhammika Leshan ORCID: https://orcid.org/0000-0001-6316-1221; Amarasiri, Mohan ORCID: https://orcid.org/0000-0002-3509-3433; Phattharapornjaroen, Phatthranit ORCID: https://orcid.org/0000-0002-4391-2929; Hurst, Cameron; Modchang, Charin ORCID: https://orcid.org/0000-0002-0739-006X; Cynthia, Bernadina; Miyanaga, Kazuhiko; Cui, Longzhu ORCID: https://orcid.org/0000-0002-8909-3885; Fernandez, Stefan; Melhem, Nada M. ORCID: https://orcid.org/0000-0002-5423-4908; Luk-in, Sirirat ORCID: https://orcid.org/0000-0003-0869-7562; Singer, Andrew C. ORCID: https://orcid.org/0000-0003-4705-6063; Ragupathi, Naveen Kumar Devanga ORCID: https://orcid.org/0000-0001-8667-7132; Htun, Thin Sandi; Sei, Kazunari ORCID: https://orcid.org/0000-0002-0684-6002; Ngamwongsatit, Natharin ORCID: https://orcid.org/0000-0001-9569-396X; Hui, Ng Sin ORCID: https://orcid.org/0000-0003-4183-4071; Shimotai, Yoshitaka; Ounjai, Puey ORCID: https://orcid.org/0000-0002-3335-6199; Kanthawee, Phitsanuruk; Tacharoenmuang, Ratana; Kauba, Alice; Eang, Chorney; Romone, Lavinia; Dharne, Mahesh; de Araujo, Juliana Calabria; Ndatuwong, Lazarus G.; Werawatte, W.K.C.P.; Chanthasiri, Traychit; Zhao, Jinxin ORCID: https://orcid.org/0000-0002-0604-3541; Mori, Hirotake ORCID: https://orcid.org/0000-0002-7988-5758; Besa, John Jefferson V. ORCID: https://orcid.org/0000-0002-5505-1855; Kurt, Özgür; Kanjanabuch, Talerngsak ORCID: https://orcid.org/0000-0002-2996-8934; Zahraei-Ramazani, Ali Reza ORCID: https://orcid.org/0000-0001-7820-3586; Higgins, Paul G. ORCID: https://orcid.org/0000-0001-8677-9454; Aoyagi, Tetsuji; Kicic, Anthony; Trowsdale, Sam ORCID: https://orcid.org/0000-0001-8907-6885; Hongsing, Parichart; Yang, Xiaonan; Wang, Yangzhong; Khatib, Aisha; Sano, Daisuke ORCID: https://orcid.org/0000-0001-8075-6972; Shibuya, Kenji; Abe, Shuichi ORCID: https://orcid.org/0000-0001-8138-5077; Hamamoto, Hiroshi ORCID: https://orcid.org/0000-0001-9315-7442. 2026 Tracking Plasmodium knowlesi through faecal DNA for monitoring zoonotic transmission in wild macaques across Southeast and South Asia. The Journal of Infectious Diseases, article jiag128. 10.1093/infdis/jiag128

Abstract
We conducted the non-invasive surveillance of Plasmodium knowlesi in wild macaques using 4,752 faecal samples collected across nine endemic countries. Parasite DNA was detected in 390 samples (8.2%), with positivity rates ranging from 1.4% to 18.4%. This provides the first field-based evidence that P. knowlesi DNA in faeces shed by macaques and present under natural conditions can be detected. These findings validate faecal sampling as a practical and scalable tool for tracking zoonotic-malaria. The results support integration into forest-runoff and rural wastewater surveillance systems, offering new opportunities for early detection of pathogens and environmental monitoring at the human–wildlife interface.
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