Six Essential oils (EO) (tea tree oil-TeTr, oregano oil-Ore, clove bud oil-Clo, thyme oil-Thy, rosemary oil-Ros, and sage oil-Sag) in Experiment 1; and different combinations of selected oils in Experiment 2, were evaluate at four doses in an in vitro microbial fermentation system using ruminal fluid from beef cattle fed a 10:90 straw: Concentrate diet. In Experiment 1, TeTr, Ore, Clo and Thy improved rumen fermentation profile in a direction consistent with better feed utilization. In Experiment 2, TeTr mixed with Thy, Ore, Thy + Ore or Clo at 200 and 400 mg/L increased the molar proportion of propionate and decreased that of acetate, and the acetate to propionate ratio. However, the size of the effect was similar to that obtained with TeTr alone, suggesting that effects were not additive. When Thy, Ore or Thy + Ore where mixed with Clo, most effects on rumen fermentation profile disappeared, suggesting an antagonistic interaction of Clo with Thy and Ore. Results do not support the hypothesis of additivity among the EO tested, and antagonistic effects of Clo mixed with Thy or Ore were demonstrated at least in a low pH, beef-type fermentation conditions.