Wannigama D.L.Amarasiri M.Sano D.Hamamoto H.Mahidol University2026-03-312026-03-312026-01-01Methods in Microbiology (2026)05809517https://repository.li.mahidol.ac.th/handle/123456789/115913Wastewater-based epidemiology (WBE) has emerged as a powerful, population-level surveillance approach for monitoring infectious disease dynamics independent of individual testing behavior. The global re-emergence of Mpox highlighted the limitations of clinic-based surveillance systems, particularly in settings affected by stigma, limited healthcare access, or rapidly changing transmission networks. Multiple studies have demonstrated that Mpox virus DNA can be detected in municipal wastewater, often preceding or paralleling reported clinical cases. This chapter reviews advances in wastewater and environmental surveillance for Mpox, with a focus on analytical methodologies, sampling strategies, laboratory workflows, quality assurance, and data interpretation. We discuss biological considerations relevant to environmental shedding, compare influent and solids-based sampling approaches, and evaluate molecular detection technologies including quantitative PCR, digital PCR, and sequencing-based confirmation. We further examine challenges related to signal normalization, inter-laboratory comparability, ethical governance, and translation of wastewater signals into public health action. Finally, we propose a practical framework for implementing sustainable Mpox wastewater surveillance programs that can be integrated into broader One Health and emerging pathogen monitoring systems.MedicineImmunology and MicrobiologyArticleSCOPUS10.1016/bs.mim.2026.02.0012-s2.0-105033279485