Simple Summary High-producing dairy cows face an increased risk of subacute ruminal acidosis, which negatively affects the general health, feed intake, and the quantity and quality of milk produced. The aim of this study was to investigate the effect of calcareous marine algae (Lithotamium calcareum)-based rumen content buffer included in total mixed ration, fed to 34 high-producing, peak lactation Holstein dairy cows (group A, n = 17; group B, n = 17). It was hypothesized that through a rumen content buffering effect, buffer would improve feeding behavior, feed intake, rumen function, and performance. Differences between the experimental and control total mixed ration in most parameters under study (feed intake, rumen content acidity, feces composition, milk yield, and milk composition) were statistically insignificant. However, statistically significant differences were noted in the length of eating and chewing epizodes (feeding time; rumination time) between groups. Feed intake was in line with this, although the difference was only numerical and statistically insignificant. This may simply mean that the buffer effects takes some time to show but persists after the buffer withdrawal. One of the reasons for not achieving a significant improvement in other parameters might be that the cows were fed a typical, well-balanced ration that did not induce acidosis.Abstract The aim of the study was to investigate the effect of calcareous marine algae (Lithotamium calcareum)-based rumen content buffer (CMA) included in concentrated feed within total mixed ration (TMR), fed to 34 peak lactation (87-144 days in milk) Holstein dairy cows, randomized into two groups (group A, n = 17; group B, n = 17), wearing collars with accelerometers, and housed a in barn with automatic feed-weigh troughs. During the first phase P1, group A received TMR with CMA (TMR-E) and group B was fed TMR without the buffer (TMR-C). For P2, the treatments in the groups were exchanged. Feed intake, feeding time (FT), rumination time (RT), milk yield, milk composition, and rumen pH were measured by barn technologies, and rumen fluid and feces composition were analyzed in the laboratory. Differences between the TMR-E and TMR-C in most parameters under study were statistically insignificant, except overall FT and RT, which differed significantly between the groups. Group A, feeding at P1 by TMR-E, exhibited higher FT and RT than Group B (202 min/cow/day vs. 184 min/cow/day, and 486 min/cow/day vs. 428 min/cow/day, respectively). The RT significantly increased after switching from TMR-C to TMR-E. This implies that the buffer effect is delayed and persists after the withdrawal. In the group of cows that received control TMR without buffer in the first phase, RT and milk protein content increased significantly in the first week after the addition of buffer.