Abstract The objective was to characterize the effects of ruminal acidosis on rumen fermentation of steers adapted to a high concentrate diet. Cattle entering the feedlot undergo a transition period where the amount of concentrates in the diet is gradually increased to mitigate the risks of ruminal acidosis. Despite a dietary adaptation, rumen acidosis still affects feedlot cattle later in the finishing phase. Ruminally cannulated steers [n = 11; body weight (BW) = 795 kg ± 54) were blocked into 2 groups based on initial BW. Before the start of the study, cattle had been consuming a finishing diet (60% dry rolled corn, 15% modified distiller’s grains, 15% corn silage, and 10% ground corn based supplement) for more than 150 d. Steers were randomly assigned to 1 of 2 treatments: control (CON) or acidosis (ACD). For the experiment, steers were fasted for 24 h and at the conclusion received the same finishing diet. Steers on ACD steers were also ruminally dosed 0.05% BW of wheat starch on h 0 and h 12 to induce acidosis. Rumen fluid was collected every 4 h for 48 h to analyze ruminal pH, ammonia, volatile fatty acids (VFA), and lactate. Fecal grab samples were collected every 8 h for 48 h to determine fecal pH. Data were analyzed with the MIXED procedure of SAS 9.4 with the effects of treatment, time, and their interaction. A treatment × hour (P < 0.01) effect was observed for ruminal pH with the ACD steers having a lower pH than CON from h 12 to 32. Fecal pH was not affected by treatment, time, or their interaction (P ≥ 0.12). Steers fed ACD had greater total VFA concentration than CON (P < 0.01). A treatment × hour interaction (P < 0.01) was observed for acetate molar proportion with CON having greater percentage acetate at h 24 through 32. Additionally, there was a treatment × hour interaction (P < 0.01) for propionate molar proportion with ACD steers having greater propionate at h 28 and 32, and then at h 40 and 44. Butyrate was not affected (P ≥ 0.58) by treatment × hour or treatment. Acetate to propionate ratio (A:P) was affected by treatment × hour (P < 0.01) with CON steers having greater A:P after h 28. Rumen ammonia and lactate concentrations did not differ (P ≥ 0.25) by treatment or the interaction with h. In steers adapted to a high concentrate diet, acidosis only altered ruminal fermentation in regards to ruminal pH and VFA concentrations.