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Background. With climate change, adverse human health effects caused by heat exposure are of increasing public health concern. Forests provide beneficial ecosystem services for human health, including local cooling. Few studies have assessed the relationship between deforestation and heat-related health effects in tropical, rural populations. We sought to determine whether deforested compared to forested landscapes are associated with increased physiological heat strain in a rural, tropical environment. Methods. We analyzed data from 363 healthy adult participants from ten villages who participated in a two-by-two factorial, randomized study in East Kalimantan, Indonesia from 10/1/17 to 11/6/17. Using simple randomization, field staff allocated participants equally to different conditions to conduct a 90 min outdoor activity, representative of typical work. Core body temperature (CBT) was estimated at each minute during the activity using a validated algorithm from baseline oral temperatures and sequential heart rate data, measured using chest band monitors. We used linear regression models, clustered by village and with a sandwich variance estimator, to assess the association between deforested versus forested conditions and the number of minutes each participant spent above an estimated CBT threshold of 38.5 degrees C. Results. Compared to those in the forested condition (n = 172), participants in the deforested condition (n = 159) spent an average of 3.08 (95% confidence interval (CI) 0.57, 5.60) additional minutes with an estimated CBT exceeding 38.5 degrees C, after adjustment for age, sex, body mass index, and experiment start time, with a larger difference among those who began the experiment after 12 noon (5.17 [95% CI 2.20, 8.15]). Conclusions. In this experimental study in a tropical, rural setting, activity in a deforested versus a forested setting was associated with increased objectively measured heat strain. Longer durations of hyperthermia can increase the risk of serious health outcomes. Land use decisions should consider the implications of deforestation on local heat exposure and health as well as on forest services, including carbon storage functions that impact climate change mitigation.