The aim of this study was to evaluate the seasonal behavior and milk yield of dairy cows of Czech Fleckvieh cattle. The subject of the monitoring was one section (housed in one quarter of barn) with 103 free cubicle beds with an average of 95 lactating dairy cows of Czech Fleckvieh cattle. In the seasons (spring, summer, autumn, winter) temperature (°C), relative humidity (%) and temperaturehumidity index (THI) were monitored. Furthermore, behavioral signs were also observed (a total of 4,940 observations): dairy cows were either lying down (3,432 observations) or standing up (1,508 observations). In the conditions that dairy cows were standing up in the cubicle, up to 585 observations were proved. If dairy cows were standing up outside of the cubicle (923 observations), they were either lying on the left side (1,924 observations) or right side (1,508). Significant seasonal influence was found out (p < 0.05) on the number of dairy cows standing up (a maximum of 410 observations in the spring, a minimum of 342 observations in the summer) and then the number of cows lying both on the left (a maximum of 519 observations in the autumn and a minimum of 444 observations in the spring) and on the right side (a maximum of 415 observations in the winter, a minimum of 320 observations in the autumn). The seasonal influence was no significant (p > 0.05) in the remaining behavioral signs. With regard to milk yield, a significant seasonal influence was proved. The highest milk yield was reached with dairy cows in spring (29.27 kg of milk) and the lowest in the autumn (24.58 kg of milk). No significant differences of milk yield were detected between behavioral signs (p > 0.05). The maximum difference of milk yield was found out up to 1.39 kg between dairy cows lying down on the left side (28.35 kg) and the dairy cows standing up in a cubicle (26.96 kg) in the winter but even this difference was not statistically significant (p > 0.05).