Differences in follicular development and repeatability of follicular growth pattern among Czech Fleckvieh (n = 20) and Holstein (n = 23) heifers were investigated. Follicular dynamics was evaluated by daily sonographic scanning during three interovulatory intervals (IOI). The mean duration of the IOI was 20.66 ± 0.32 days, no differences between breeds. The proportion of the non-alternating pattern (54%) was nearly the same as that of the alternating pattern (46%). Comparing IOI, 44.2% of the heifers exhibited 3 waves and 55.8% 2 waves of follicular growth. The ratio of 3- to 2-wave heifers was about the same in the Holstein breed (11/12); in Czech Fleckvieh 2-wave cycles slightly dominated (8/12). In Holstein heifers, the first follicular wave occurred 0.92 ± 0.15 days after ovulation, and the first wave in 2-wave Czech Fleckvieh heifers appeared later (P < 0.05), 1.83 ± 0.3 days. In conclusion, we found a similar pattern of ovarian follicular dynamics in Czech Fleckvieh and Holstein heifers.