This study was designed to determine the effects of dietary propylene glycol (PG; Propypact®, DIFAGRI, France) on blood metabolites, metabolic and reproductive hormones and follicular growth in 10 dairy heifers. Treatments consisted of (1) 1.1 kg of sugar beet pulp (Control), (2) 150 g PG (PG150) and (3) 300 g PG (PG300). Each heifer received the three treatments in different randomized orders. A standard hay/concentrate diet, formulated to allow a daily liveweight gain of 900 g/day, was given at 8:00 and the dietary treatments were given at 16:00 from Days 1 to 13 of the oestrous cycle following induced oestrus (Day 0). Oestrus induction treatment consisted of a subcutaneous 3 mg norgestomet implant inserted for 9 days combined with GnRH treatment (i.m.) at implant insertion. Two days before implant removal, 500 µg cloprostenol was administered i.m. Blood samples were collected by jugular venipuncture every two hours for 24 hours on Days 0 and 13 to measure plasma insulin, glucose, ?-hydroxybutyrate (BHB) and urea concentrations. Blood samples were also collected to measure insulin-like growth factor-1 (IGF-1), oestradiol, progesterone concentrations on Days 2, 6, 9 and 12 and AMH (Anti-Müllerian hormone) on Days 0, 2, 6, 9 and 12. On Days 2, 6, 9 and 12 ovarian follicular growth was evaluated; the total number of follicles and their diameters were recorded and classed (2–3 mm, 4–7 mm, and>8 mm). Results were analysed by repeated-measures ANOVA. There were no treatment, day and interaction effects on average urea concentrations while there were some differences between Days 0 and 13 for insulin, glucose and BHB. Insulin and glucose concentrations were higher on Day 13 compared to Day 0 and the opposite was observed for BHB. There were treatment, time and interaction effects on glucose and BHB concentrations measured over 24 h on Day 13; glucose concentrations were higher (P<0.05) at 4:00, 8:00, 12:00, 16:00 and 20:00 h, whereas BHB concentrations were lower (P<0.05) at 20:00 and 22:00 h in the PG300 group compared to the control and PG150 groups. There were treatment, day and interaction effects on IGF-1 and progesterone concentrations, and the number of small follicles. PG150 resulted in higher progesterone concentrations on Days 9 and 12, and more small follicles on Day 2 compared to Control. AMH concentrations were unaffected by day of oestrous cycle and dietary treatment. However a negative correlation was observed between pre-PG distribution insulin and AMH (r=?0.47, P<0.05). These results indicate that short-term dietary PG supplementation affects circulating concentrations of metabolites and metabolic hormones, increases progesterone concentrations and the number of small follicles. Propylene glycol supplementation may be effective in improving oocyte production when combined with hormonal treatments to stimulate follicular growth for superovulation or ovum-pick up.