A study was carried out to understand the impact of cultivation of Bt cotton on nutrient availability and soil physico-chemical properties by collecting geo-referenced soil samples from fields of farmers of Warangal district. Sampling was done from fields under Bt cotton for more than 8 years, from fields under Bt cotton for 2-5 years and from non Bt cotton fields covering 11 mandals of district. From the survey it was evident that farmers of this region were using very high doses of fertilizers viz., 333 kg N, 97 kg P2O5, 142 kg K2O and 17 kg S per ha for cultivation of Bt cotton. Results clearly indicated that cultivation of Bt cotton either for prolonged period (> 8 years) or short term (2-5 years) did not bring out any significant change in soil pH, EC, CEC and organic carbon content when compared to non Bt cotton cultivation. Except available phosphorus, the availability of nitrogen, potassium, sulphur and micronutrients did not show significant changes. Available phosphorus status was found to be significantly high in soils under >8 years of Bt cotton and multiple years of cultivation of Bt cotton did not affect the functional bacterial population, microbial biomass carbon (MBC) and enzyme activity.