Understanding variations in the reproductive cycle of commercial and ecologically important bivalve species is essential to address fisheries management and climate oscillation issues. Spatial and temporal trends in the reproductive cycle of Callista chione were analysed using standard histological procedures and gonadosomatic index (GSI) for two consecutive years at two coastal sites in the Adriatic Sea. Reproductive output and fecundity were analysed quantitatively to gain better insight into the seasonal energy balance with respect to gametogenesis. A seawater temperature difference of ~4°C between the two sites likely influenced the onset and duration of spawning. At Cetina (milder temperatures), spawning was prolonged and occurred between spring and July, whereas at Pag there was only one main spawning peak in July. Both histology and GSI showed similar trends, indicating GSI to be a reliable method for analysing the main characteristics of the reproductive cycle. Conversely, the use of the body mass index (BMI) as an indicator of gonad development should be addressed carefully. Intraspecific variation in energy allocation between populations suggested life strategies modified to different habitat conditions. A higher reproductive investment and output in the Pag population directed gonadal resorption to ensure the generation of a new cycle, whereas the higher fecundity at the Cetina site indicated the production of gonads from newly available food. These results suggest successful physiological acclimatization to recent increasing temperatures.