We examined the effects of three maternal diets (very high fat (VHF), low fat (LF), and control (Purina 5015)) on serum steroids, free fatty acids (FFA), and vaginal pH in National Institutes of Health Swiss mice. Females were fed (VHF, nZ33; LF, nZ33; 5015, nZ48) from 4 to 16 weeks of age. Following breeding, female serum was collected at 0 . 5 (pre-implantation, early diestrus) or 8 . 5 (post-implantation, mid-diestrus) days post-coitus (dpc). The serum concentrations of 17b-estradiol, testosterone, progesterone, and FFA were analyzed at both collection points, and vaginal pH at 0 . 5 dpc. Striking differences in steroids and FFA were observed at 0 . 5 dpc among the groups. Estradiol was higher in the VHF (14 . 1G3 . 0 pg/ml), compared with LF mice (5 . 2G2 . 3 pg/ml; P%0 . 05). In contrast, 0 . 5 dpc testosterone was lower in the VHF (10 . 5G3 . 0 pg/ml) versus the LF group (32 . 7G8 . 4 pg/ml; P%0 . 05). At 8 . 5 dpc, progesterone was higher in the VHF (89 . 6G6 . 7 ng/ml) versus the 5015 group (60 . 1G4 . 9 ng/ml; P%0 . 05). VHF mice had higher FFA concentrations at 0 . 5 dpc (1 . 0G0 . 2 mmol/l) than LF and control mice (0 . 5G0 . 1 and 0 . 6G0 . 1 mmol/l respectively; P%0 . 05). At 8 . 5 dpc, VHF females had higher serum FFA (0 . 8G0 . 1 mmol/l) than LF and control females (0 . 4G0 . 1 and 0 . 6G0 . 1 mmol/l; P%0 . 05). Mean vaginal pH of VHF females (6 . 41G0 . 09) was lower than 5015 females (6 . 76G 0 . 10; P%0 . 05). These diet-induced alterations in serum steroid and FFA concentrations might affect several reproductive processes, including preferential fertilization by one class of sperm over the other and sex bias in pre-and post-implantational embryonic development.