Fifty-seven cycling buffalo cows of the river type were treated with two doses of 0.5 mg cloprostenol intramuscularly given 11 days apart. Each animal was inseminated twice at 72 and 96 hours after the second injection of cloprostenol. The first service conception rate diagnosed by rectal palpation at 90 days was 38.6 per cent. At the time of insemination the cervix was easily penetrable on both days in only 39 (68.4 per cent) of the animals. They were inseminated at or beyond the internal cervical os, while the others were inseminated in the cervical canal. There was a marked difference in conception rate between those receiving deep inseminations (48.7 per cent) and the others (16.7 per cent). In relation to the interval from calving to insemination the conception rates for those which had calved 60 to 90, 90 to 120 and 120 to 150 days earlier were 16.6, 36.4 and 55.5 per cent respectively. The use of cloprostenol treatment and fixed-time insemination is a useful method of overcoming the problem of oestrus detection in buffaloes. Acceptable levels of fertility can be obtained in those animals which have a sufficiently relaxed cervix to permit semen deposition at the internal os, provided the interval from calving to insemination is more than 90 days.