Lactating cows and nulliparous heifers are in distinctive and unique physiological conditions when they are approaching the planned time of breeding, at approximately 60 days in milk and 13-15 months of age, respectively. This study aimed to profile the metabolic milieu in heifers (N = 14) and lactating cows (N = 15) in the weeks leading up to planned time of breeding. All cows were followed for a period of 15 weeks, from 3 weeks pre-calving to 12 weeks post-calving, while heifers were monitored for a period of 4 weeks leading up to the tentative week of breeding (pre-breeding period). For data analysis, we further divided cows into primiparous (N = 8) and multiparous (N = 7) cows owing to the significant difference in their milk yield. Assessment of reproductive performance showed that primiparous and multiparous cows tended to have lower pregnancy rates compared to heifers (P < 0.1). Plasma concentrations of β-hydroxybutyric acid were about 2-fold higher in multiparous cows than those of heifers in the week leading up to planned time of breeding (P < 0.05). Total bile acid levels during the pre-breeding period were higher in all lactating cows compared to heifers (P < 0.05) and glucose levels were lower in lactating cows (P < 0.05). Triglyceride concentrations were lowest in multiparous cows compared to both primiparous cows and nulliparous heifers (P < 0.05). In addition, lactating cows had higher concentrations of total-cholesterol and the high-density lipoprotein and low-density lipoprotein compared to heifers (P < 0.05). Conversely, concentrations of very low-density lipoprotein were lower in multiparous cows than primiparous cows and nulliparous heifers (P < 0.05). There were no differences in plasma glutathione levels, as measured by liquid chromatography-tandem mass spectrometry, between the groups, but the ferric reducing ability of plasma was higher in lactating cows compared to heifers (P < 0.05). These data establish the differences in the profile of metabolic and oxidative markers during the period approaching planned time of breeding in lactating cows compared to nulliparous heifers. As certain metabolites in the plasma have been shown to be represented in the ovarian follicular microenvironment, the unique profiles may influence reproductive performance in dairy cattle in different physiological stages.

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