A study was conducted to test the hypotheses that exogenous melatonin treatment of 11-13 month-old red deer hinds: (1) advances the timing of first ovulation, (2) increases the proportion of individuals attaining puberty at ∼16 months of age, and (3) reduces the live-weight threshold for attainment of first pregnancy. A total of 3901 rising-2-year-old (R₂) hinds within two herds (A and B) across two years either received single melatonin implants on two occasions in summer (n=1399) or were untreated controls (n=2502). Hinds were joined with stags from mid January to mid May, and were subjected to real-time rectal ultrasonography in early June to assess pregnancy status (proxy for puberty attainment) and foetal age for conception date assignment. Live-weights were recorded for each hind in January (12 months of age) as a proxy for weight at puberty. Melatonin treatment of hinds was associated with a significant advancement in mean conception dates in both herds in both years (P<0.05), with a cohort difference in mean dates between treated and control hinds ranging from 9 to 17 days. Analysis of the temporal distribution of conception dates for each cohort revealed bi-modal or tri-modal patterns of conception indicative of conceptions to first or subsequent ovulations (oestrous cycles). Across all cohorts, melatonin treatment was associated with higher conception rates to first ovulation (P<0.05) resulting in greater overall synchrony of conceptions. Regression analysis demonstrated a significant negative slope for conception date against live-weight (P<0.001), but there was no evidence that this slope varied with treatment, herd or year (P>0.05); for every 10kg increase in live-weight conception date was advanced by an average of 1.3 days. In Herd A, melatonin treatment was associated with significantly higher pregnancy rates in both years (90.3% vs. 78.0% in Year 1 and 84.4% vs. 57.1% in Year 2; P<0.05). The principle effect of melatonin treatment was to increase the pregnancy rate of hinds of low body-mass. In Year 1, at 60kg live-weight a logit regression model indicated a pregnancy rate of 52% for untreated hinds and 83% for treated hinds. At 105kg the rate for both cohorts was 90%. In Herd B, melatonin treatment was associated with higher conception rates in both years but these differences were not significant following correction for slight differences in mean live-weight (P>0.05). The study has demonstrated that factors influencing puberty attainment in R₂ red deer hinds can vary between populations. In Herd A, in which body mass of hinds immediately prior to their first potential breeding season may have been the principle limiting factor, melatonin treatment appears to have instigated the pubertal process in hinds that would otherwise be of insufficient body mass.