When two injections of 500 microgram cloprostenol are given 11 days apart to unselected cycling heifers the onset of oestrus behaviour is significantly earlier, and more compact, after the second injection than after the first. The time from treatment to the onset of oestrus following this second injection was not significantly different from that obtaining in animals given a single injection on day 8 of the oestrous cycle. It is suggested that the more precise onset of oestrus after the second of two cloprostenol injections is due to the majority of animals being at a comparable stage of the oestrous cycle when the treatment is given. The earlier oestrus after two cloprostenol injections may be due to the greater number of heifers at days 7, 8 and 9 of the oestrous cycle at the time the second injection is given.