Four groups of 20 Simmental cross heifers were actively immunized against human serum albumin (control, (C), androstenedione (A), a 30 amino acid synthetic fragment of porcine inhibin alpha (I) or A and I in combination (AI). At the time of the third and final antigen injection and again 2 weeks later, all animals were injected with synthetic prostaglandin F2α (PG) to synchronize oestrus. Concentrations of LH, FSH, insulin-like growth factor-1 (IGF-1) and inhibin (C and A heifers only) in serum samples collected twice daily during 3 days before the second PG injection and at 15-min intervals from 24 to 32 h after PG injection were determined. Two weeks later, all heifers were administered PG and ovariectomized 24 h later. Ovarian follicles were dissected from the stroma, measured and cultured in medium 199 for 2h at 37°C. Numbers of small (3·0 to 7·9 mm diameter) and large (>8·0 mm diameter) follicles were not significantly altered by any of the immunization treatments. Intrafollicular concentrations and secretion rates of oestradiol, testosterone, progesterone and inhibin, determined in vitro, were not affected by immunization. Intrafollicular IGF-1 concentrations were not affected; this hormone was not detectable in the culture medium. Intrafollicular concentrations of oestrogen and testosterone and secretion rates of all three steroids were greater (P < 0·05) in animals which had regressing corpora lutea at the time of ovariectomy. Of the animals that had dominant follicles, one of 17, one of 13, four of 19 and six of 15 in the C, A, I and AI groups, respectively, had two or more. Only the difference between AI and C cows in the incidence of dominant follicles was statistically significant (P < 0·05). Mean circulating concentrations of LH, and inhibin (C and A groups only) and mean LH pulse frequency and pulse amplitude were unaffected by treatment but FSH concentrations were higher in A (P < 0·001) and lower in AI and I heifers (P < 0·001) compared with C animals while IGF-1 concentrations were lower in A heifers (P < 0·001) and higher in AI heifers (P < 0·05) compared with C heifers. It is concluded that immunization against inhibin, either alone or in combination with immunization against androstenedione increased the incidence of multiple dominant follicles in heifers but did not increase their ovulation rate.