We conducted the noninvasive surveillance of Plasmodium knowlesi in wild macaques using 4752 fecal samples collected across 9 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 feces shed by macaques and present under natural conditions can be detected. These findings validate fecal 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. This study validates noninvasive fecal sampling for tracking Plasmodium knowlesi in wild macaques, providing a scalable tool for early pathogen detection and environmental malaria surveillance.

Foreign