The present study reports the pulsatile secretion of gonadotrophins and 17β-oestradiol (OE2) on the day of oestrus and at the mid-luteal phase of the oestrous cycle in mithun (Bos frontalis). The frequency of luteinising hormone (LH) and follicle-stimulating hormone (FSH) pulses was found to be greater (P < 0.05) on the day of oestrus than at the mid-luteal phase. In contrast, the amplitude of the LH and FSH pulses was greater (P < 0.01) at the mid-luteal phase than on the day of oestrus. A synchronised (P < 0.01) LH and FSH secretion pattern was found only at the mid-luteal phase. A pulsatile secretion pattern for OE2 in the peripheral circulation was evident for both phases of the oestrous cycle. The frequency of the OE2 pulses did not differ significantly in different phases of the oestrous cycle. In contrast, the amplitude of the OE2 pulses and the basal OE2 concentration were found to be greater (P < 0.01) at the mid-luteal phase than on the day of oestrus. A synchronised (P < 0.01) LH and OE2 secretion pattern was observed in both phases of the oestrous cycle. In contrast, a synchronised (P < 0.05) FSH and OE2 secretion pattern was found only on the day of oestrus. In conclusion, a different pattern of LH and FSH secretion was observed in both phases of the oestrous cycle, mainly on the day of oestrus, which indicates a differential regulatory mechanism of LH and FSH release. In addition, as in cattle, OE2 exerts a positive feedback on LH and FSH release on the day of oestrus and on LH release at the mid-luteal phase of the oestrous cycle in mithun. Also, as in cattle, the greater basal plasma OE2 concentration and increased amplitude of OE2 pulses exert a negative feedback on FSH release at the mid-luteal phase of the oestrous cycle.