Chemical Characterization and Rumen-Modulating Effects of Pinus sylvestris Essential Oil: In Vitro and In Vivo Study

Conifer-derived essential oils have gained attention as versatile natural additives with potential applications in animal production, including influencing microbial processes and supporting environmental sustainability. This study aimed to characterize the chemical composition of selected conifer essential oils (EOs), evaluate their antimicrobial activity against rumen microorganisms in vitro, and assess the effects of Pinus sylvestris essential oil on rumen fermentation and methane production under in vitro and in vivo conditions. EOs from Thuja occidentalis, Cupressus sempervirens, Juniperus communis, Picea mariana, Pinus sylvestris, and Pinus pinaster were analyzed by GC–MS, and their inhibitory activity against selected rumen bacteria was determined by MIC and IC50 assays. Based on these results, P. sylvestris oil was selected for fermentation experiments. Ninety-two volatile compounds were identified, with monoterpenes as the dominant constituents and α-pinene as the major compound in P. sylvestris oil. In vitro, P. sylvestris oil influenced fermentation in a dose-dependent manner without affecting ruminal pH. In vivo, ruminal pH, ammonia-related parameters, and total VFA concentration were not significantly affected by treatment, whereas several variables showed a significant effect of time. Temporal changes in VFA profiles suggested a transient adaptation of ruminal fermentation. Methane concentration was significantly (p < 0.01) reduced by Pinus sylvestris essential oil supplementation, with a decrease of approximately 28.7% after 14 days. These findings indicate that P. sylvestris EOs may serve as a promising natural modulator of rumen fermentation, although further studies are needed to optimize dosage and confirm long-term effects.