Rats were immersed for 15 min in a forced swim test in cylinders filled with water to a depth of 20 or 35 cm. The following day, they were reimmersed for a 5-min test period at one of these two depths. Water immersion, regardless of water depth, resulted in increased serum lactate and corticosterone levels, reduced serum carbon dioxide and potassium levels, a metabolic acidosis, and an increase in the phosphorus/potassium ratio. Testing at the greater water depth resulted in lower immobility times, higher lactate and lower carbon dioxide levels, a greater metabolic acidosis, and a higher phosphorus/potassium ratio than testing at the shallower depth. Water depth did not significantly affect corticosterone or potassium responses. None of the interactions between day 1 and day 2 depth was significant. The correlation between immobility time and corticosterone was very low, whereas the correlation with lactate, carbon dioxide, anion gap, and the phosphorus/potassium ratio were high. The inference from these results is water depth at time of testing affects the rat's behavioral and physiological reactions in the forced swim test and prior exposure to water at any depth does not.