Four females were studied during a 10-day heat acclimation regimen consisting of 2 h per day of stationary cycle riding (means = 46% VO2 max) in a hot environment (40 degrees C, 50% RH) to determine the effects of heat acclimation on changes in plasma volume and vascular protein both at rest and during exercise. Changes in plasma volume were calculated from hematocrit and hemoglobin measurements. Plasma volume decreased during each 2-h session on baseline days through heat day 6 (-4.9% to -1.2%). On heat days 8 and 10, there was an expansion of plasma volume during the session (1.4% and 0.8%). This hemodilution was significantly different (P less than 0.05) from the hemoconcentration on baseline through heat day 4. The gain of protein during the 2-h session was not significantly different on any of the heat days from that on baseline days. With acclimation of females to work in the heat, there is an increased ability to maintain the fluid of the vascular compartment during a 2-h exercise session in the heat.