This study examined the effects of dry glycerol supplementation on follicular growth, post-partum interval to first ovulation, concentration of serum metabolites and hormones related to fertility, body condition score (BCS) and body weight (BW) in primiparous Holstein dairy cows. Sixty primiparous Holstein dairy cows were randomly assigned to two groups (control: n = 30 and glycerol supplemented: n = 30). Dry glycerol (250 g/day/cow) was fed as a top dressing to the common lactating total mixed ration (TMR) from parturition to 21 days post-partum. Ovaries were examined four times using ultrasonography on days 13, 19, 25 and 36 post-partum to determine ovarian follicular growth. Concentration of serum metabolites and hormones was determined weekly. Body condition score was evaluated weekly from weeks 1 to 5 after parturition, and BWs were recorded three times on days 1, 11 and 21 during the experimental period. The cows fed dry glycerol had more large follicles (p < 0.0001) and corpora lutea (CL) (p = 0.02) compared with the control cows. Days to the first ovulation (p = 0.06), days to first oestrus (p = 0.05), services per conception (p = 0.06) and days open (p = 0.004) were positively affected by dry glycerol supplementation. Serum concentration of glucose and insulin was higher in dry glycerol-supplemented cows (p = 0.1; p = 0.06, respectively). Feeding glycerol had no effect on mean serum concentrations of β-hydroxybutyrate, non-esterified fatty acids and IGF-1 during the experimental period. However, significant differences were observed at concentration of BHBA and IGF-1 (p = 0.02 and p = 0.04, respectively) between two groups on day 21 after calving. The cows in the glycerol-fed group had higher serum progesterone concentrations on days 33 (p = 0.007) and 36 (p = 0.004) after calving. Supplemented cows had lower body condition loss during weeks 1–5 after calving compared with the control cows (0.34 vs 0.41 BCS). In week 13 post-partum, the proportion of cycling cows was 83.3 and 69.9% for those which received supplemented or non-supplemented diet, respectively. These results demonstrated that feeding dry glycerol as a glucogenic supply may be useful to improve negative energy balance and reproductive efficiency in young cows which calve with high requirement of energy.