The objective of the study was to evaluate the effects of three nutritional treatments, which differed in energy level and source, on preovulatory follicles, number of follicles and oestrous activity in dairy cows. Twenty two Holstein multiparous cows from the Elsenburg herd were used in this study. After parturition, cows were kept on kikuyu-ryegrass pastures on an ad libitum basis, and allocated to various levels and types of concentrate supplements, which differed in starch and fat contents. The control group received 7 kg/day of a control concentrate, and the treatment groups each received 12.6 kg/day of concentrate. The concentrates contained high starch-low fat (HSLF) and high starch-low fat/low starch-high fat (HSLF-LSHF) levels. The supplement in treatment HSLF was a glucogenic concentrate using maize as the energy source. The supplements in treatment HSLF-LSHF were a combination of a glucogenic concentrate, which was offered for the first 60 days in milk (DIM), similar to treatment HSLF, followed from 61 DIM by a lipogenic concentrate using wheat bran and calcium (Ca) salts of long-chain fatty acids as the energy sources. At 80 ± 10 DIM, cows were synchronized with an Ovsynch protocol without being inseminated before the ultrasonography observation. While they were detained in a shaded neck clamp, cows were assessed individually with an ultrasound scanner every three days for ovarian measurements and follicular activity until the subsequent oestrus. Results showed that ovarian and follicular measurements and the numbers of follicles in various follicle size classes were similar between nutritional treatments. However, the total ovarian follicular counts were significantly higher in cows that received the HSLF and HSLF-LSHF treatments, compared with their counterparts in the control group (i.e. 7.23 ± 0.22, 7.21 ± 0.14 and 6.53 ± 0.19, respectively), through possible improvement in nutritional status. Further research is required to investigate various energy levels and sources that enhance the viability and the quality of the oocyte ovulating from the dominant follicle and improve the intensity and length of the oestrous expression in dairy cows.