Diurnal variations of progesterone, testosterone, and androsta-1,4-diene-3,17-dione in the rumen and in vitro progesterone transformation by mixed rumen microorganisms of lactating dairy cows

Five Holstein lactating dairy cows fed 5 total mixed rations (TMR) with different forage combinations were used in a 5 × 5 Latin square design to investigate diurnal variations of progesterone (P4), testosterone, and androsta-1,4-diene-3,17-dione (ADD) concentrations in the rumen. Meanwhile, different P4 inclusion levels [0 (control), 2, 20, 40, 80, and 100 ng/mL in culture fluids] were incubated in vitro for 6, 12, 24, 36, 48, and 72 h together with rumen mixed microorganisms grown on a maize-rich feed mixture (maize meal:Chinese ryegrass hay = :1) with an aim to determine microbial P4 transformation into testosterone and ADD. Ruminal P4, testosterone, and ADD concentrations of lactating dairy cows were greater in the TMR with forage combination of corn silage plus alfalfa hay or Chinese wild ryegrass hay than the TMR with the corn stover-based forage combination. The diurnal fluctuation pattern showed that P4, testosterone, and ADD concentrations in the rumen were greater at nighttime than daytime and peaked 6 h after feeding in the morning or afternoon. The in vitro batch cultures showed that the P4 elimination rate was highest at the P4 addition of 20 ng/mL and declined with the further increased addition of P4. The treatments after dosing P4 exhibited a shorter time than the control group until half of the initial P4 inclusion was eliminated (i.e., half time), and the lowest half time (1.46 h) occurred at the P4 addition of 20 ng/mL. In summary, the ruminal steroids concentration was affected by forage type and quality, and the rumen microorganisms exhibited great ability to transform P4 into testosterone and ADD, depending on incubation time and initial P4 addition level, suggesting that the host might affect the metabolism of its rumen microorganisms via the endogenous steroids.