Tracking <i>Plasmodium knowlesi</i> Through Fecal DNA for Monitoring Zoonotic Transmission in Wild Macaques Across Southeast and South Asia

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.