Negative energy balance (NEB) occurs in early postpartum dairy cows as a normal process when the energy demand at the onset of lactation exceeds feed energy intake. A poor response to NEB leads to greater blood concentrations of nonesterified fatty acids (NEFA) and BHBA, and also lesser blood glucose concentration. The objective was to determine whether blood BHBA and glucose concentration early postpartum affected pregnancy establishment after 3 inseminations in dairy cows. Blood samples were collected from lactating Holstein dairy cows (n = 204) twice weekly for the first 4 wk postpartum (n = 8 samples per cow). Blood BHBA and glucose were measured cowside by using a hand-held meter (Precision Xtra; Abbott Diabettes Care, Alameda, CA). Body weight (BW) and body condition score (BCS) were measured at calving, 2 wk, and 4 wk. Data were analyzed using the GLIMMIX, GLM, and MIXED procedures of SAS. The mean (SD) d postpartum of sample collection was 2.1 (1.3), 5.6 (1.5), 9.2 (1.4), 12.7 (1.6), 16.3 (1.6), 19.7 (1.6), 23.4 (1.6), and 26.8 (1.7) for samples 1 to 8. There were 171 cows with pregnancy data and 127/171 (74%) were pregnant after 3 inseminations. The blood BHBA concentration for sample 1 to 3 affected whether the cow became pregnant postpartum (lesser BHBA favorable toward pregnancy). The BHBA for non-pregnant and pregnant (respectively) was 0.91 ± 0.07 and 0.73 ± 0.04 (sample 1; P < 0.029), 0.92 ± 0.07 and 0.76 ± 0.04 (sample 2; P < 0.043) and 0.76 ± 0.04 and 0.68 ± 0.02 (sample 3; P < 0.08). The BHBA concentration for samples 4 to 8 did not affect pregnancy. Greater blood glucose concentration for sample 2 tended to be associated with pregnancy (51.3 ± 2.2 and 55.3 ± 1.3 for nonpregnant and pregnant; P < 0.11). Glucose in sample 1 and 3 to 8 were not associated with pregnancy later postpartum. BW loss and BCS loss postpartum did not affect pregnancy. In conclusion, BHBA concentrations within the first 10 d postpartum affected pregnancy outcome later postpartum.