A 3-minute step test provides a feasible, low cost procedure with minimal risk to the participant for assessing physical fitness compared to maximal exercise testing. Test assessing the autonomic function (sympathetic wing) evaluates the cardiovascular reflex like increased blood pressure triggered by performing specific stimulating maneuvers such as Isometric Hand Grip Test (IHGT). We assessed & categorized the fitness level using a 3 min step test and its association with sympathetic activation during IHGT in university level students with sound health. There is increasing evidence of an association between Physical Fitness and academic performance. Two hundred consenting healthy students studying at Pokhara University (largest in western region of Nepal) aged 17-25 years underwent sub-maximal exercise as per the protocol of 3 min-step test given by Young Man's Christian Associations (YMCA) to categorize as good & poor being based on recovery heart rate. Each groups undertook IHGT & changes in Diastolic Blood Pressure (DBP) were noted. About two third of the study participants were female and 70% of total students were of poor fitness level. Fitness level and sex was significantly associated (P=0.041, OR=1.92, CI=1.024-3.606). More of the male participants had good fitness in comparison to that of the female participants (38.9% vs 24.8%). Differences in Diastolic Blood Pressure during IHGT and at rest (assess sympathetic activation during stress) was significantly associated with fitness level (P<0. 001, OR=1.32,. Majority of university level students had poor fitness with high proportion in female. Students with good fitness level had good sympathetic activation to physical stress as compared to those with poor fitness level. Physical fitness (Cardiorespiratory fitness) is proven to induce angiogenesis in the motor cortex and increases blood flow, improving brain vascularization which could affect cognitive performance. So measuring physical fitness of university students at the time of admission or during the study period could help to predict their academic achievements through their reasoning function and ability of coping various stresses that they might encounter during academic journey. Those with poor physical fitness would be advised to improve the fitness by promoting physical activity.