The effects of single versus twice daily short term heat acclimation on heat strain and 3000m running performance in hot, humid conditions, Journal of Thermal Biology, http://dx.doi.org/10. 1016/j.jtherbio.2016.01.001 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting galley proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. .min -1 2peak , once daily for 4d (SDHA) and twice daily for 2d (TDHA), in 35°C, 60% relative humidity (RH), and once daily for 4d (CON) in 21°C, 40% RH. Participants completed a pre-and post-intervention 5 km treadmill run trial in 30°C, 60% RH, where the first 2 2peak and the final 3 km was selfpaced.No statistically significant interaction effects occurred within-or between-groups over the 2-4d intervention. While within-group differences were found in physiological and perceptual measures during the fixed intensity trial post-intervention, they did not statistically differ between-groups. Similarly, TDHA (-36±34s [+3.5%]) and SDHA (-26±28s [+2.8%]) groups improved 3 km performances (p=0.35), but did not differ from CON (-6±44s [+0.6%]). This is the first study to investigate the effects of HA twice daily and compare it with traditional single session per day STHA. These STHA protocols may have the ability to induce partial adaptive responses to heat stress and possibly enhance performance in environmentally challenging conditions, however, future development is warranted to optimise the administration to provide a potent stimuli for heat adaptation in athletic and military personnel within a rapid regime.