Microplastics (MPs) have emerged as a significant environmental threat, infiltrating livestock systems. This study presents the first in vitro investigation of the effects of low-density polyethylene (LDPE) MP contamination on rumen fermentation dynamics and feed utilization in a simulated ruminal digestive system. Concentrate feed was incubated in buffered rumen fluid collected from lambs, supplemented with LDPE MPs at concentrations of 3.3 g/L and 6.6 g/L and compared to the concentrate incubated in the buffered rumen fluid without MP contamination. The results demonstrate that both levels of LDPE MPs significantly altered rumen fermentation dynamics by reducing asymptotic gas production by 11% and 15% and increasing the constant rate of gas production by 16% and 19% at low and high addition levels, respectively, compared to the control. However, the early-stage fermentation dynamics remained unaffected. Furthermore, both levels of LDPE MPs reduced rumen protozoal populations (20% and 23%) and ammonia-nitrogen levels by 11% at both of addition levels. Despite these disruptions, rumen pH remained unaffected. Increasing the addition level of LDPE from 3.3 to 6.6 g/L did not exacerbate the disruptions. The results of this study highlight the potential risks posed by LDPE MPs in ruminal nutrition. Further in vivo investigations are essential to validate these findings and assess their impact on animal performance.