Summary Chronic stress affects brain areas involved in learning and emotional responses through modulation of neurotropic factors or neurotransmitters. Therefore, we investigated the role of exercise and thiamine supplementation on spatial memory and on brain-derived neurotrophic factor (BDNF) and acetylcholine (Ach) content in the hippocampus of the stressed animals. Male Wistar rats were randomly assigned to 4 groups (8 rats/group): control group; stress group; swimming and stress group; and thiamine and stress group. All animals were assessed by a T maze for spatial memory or open fi eld test for locomotion and anxiety. BDNF and Ach were estimated in the hippocampus. Chronic immobilization stress resulted in a signifi cant decrease in BDNF and Ach levels in the hippocampus and impairment in spatial memory functions and decreased basal activity. However, either swimming training or thiamine intake for 30 d was proved to induce a signifi cant increase both in BDNF and Ach in conjunction with improved performance in the T maze, marked anxiolytic effect and enhanced ambulation in the open fi eld test, as compared to the stress group. Interestingly, swimming-exercised rats showed signifi cantly higher levels of BDNF versus thiaminereceiving rats, while thiamine-receiving rats showed higher locomotor activity and less freezing behavior in the open fi eld test compared to the swimming group. It was concluded that decreased BDNF and Ach after stress exposure could be a mechanism for the deleterious actions of stress on memory function; swimming exercise or vitamin B 1 supplementation for 30 d was a protective tool to improve coping with chronic stress by modulating BDNF and Ach content along with enhancement of memory functions and motor activities.