The content of Ca in milk exceeds the typical saturation level of Ca salts, which is necessary for neonate growth. This calcium is distributed between the casein micelles in the colloidal and aqueous phases. Information on the properties of calcium activity in the aqueous phase is limited compared with that on the properties of bound or sequestrated calcium. The objectives of this study were to evaluate the changes in calcium activity in fresh milk using an ion-selective electrode and to assess the relationship between calcium activity and milk production in hot season. Milk samples collected from 10 cows at the National Agricultural Research Center for Kyushu Okinawa Region in June to October (Min-Max: 7.2-$35.2^{circ}C$, 24.3-100% RH) were analyzed on total calcium concentrations and calcium activity. We observed that the rectal temperature of the cows increased according to elevation of ambient temperature but that the pH of the collected milk ($6.61{pm}0.01$ (Mean${pm}$SEM)) was not significantly influenced by rectal and ambient temperature. Total calcium concentrations and calcium activity in fresh milk decreased in July (Min-Max: 21.1-$33.5^{circ}C$, 48.9-100.0% RH) compared with the values after August (Min-Max: 18.1-$35.0^{circ}C$, 26.5-96.2% RH) (p<0.05); however, there was no significant correlation between the two parameters. The ratio of calcium activity to total calcium concentration decreased after August compared with the values in June and July (p<0.05). The calcium activity in fresh milk was positively correlated with milk yield (r = 0.45, p<0.01) and negatively correlated with milk lactose content (r = -0.53, p<0.01). These results suggest that the calcium activity in milk could be affected by ambient temperature and might be associated with milking production in hot season.