Soil enzyme activities provide unique biochemical assessment of soil function as a good indicator of soil fertility which can be altered due to the profusion of fluoride in soil and seasonal change. Seven sites were chosen in the fluoride affected area of Nasipur, Birbhum district, West Bengal, India, to make a comparative assessment of the seasonal changes of enzyme (urease, amylase, cellulase and invertase), fluoride content, physicochemical characteristics along with the availability of microbes in the soil with control.