Effects of Plant Essential Oils and Their Combinations on In Vitro Ruminal Fermentation Characteristics

This study aimed to evaluate the effects of eight plant essential oils on in vitro ruminal fermentation. The oils tested were verbena (VE), cinnamon (CE), rosemary (RE), Torreya grandis (TG), ginger (GE), fennel (SF), thyme (TE), and oregano (OE). Each was supplemented at five graded doses (0, 0.75, 1.5, 3, and 6% of the substrate DM) during 24 h incubations. The results demonstrated that adding 3% CE significantly increased the in vitro dry matter degradation rate (IVDMD) while enhancing microbial protein (MCP) synthesis efficiency and acetate concentration. The addition of 1.5% SF increased MCP and total volatile fatty acid (TVFA) concentrations without affecting ruminal ammonia nitrogen levels, and 0.75% TE enhanced nitrogen utilization efficiency while maintaining primary fermentation pathways and ammonia nitrogen levels. Although 0.75% OE did not alter overall fermentation intensity or nitrogen metabolism, it specifically regulated volatile fatty acid metabolic pathways, significantly promoting butyrate production. Through preliminary screening, four essential oils (CE, SF, TE, OE) that positively influenced rumen fermentation were selected and combined into five distinct blends: T1 (CE + SF + TE), T2 (CE + SF + OE), T3 (CE + TE + OE), T4 (SF + TE + OE), and T5 (CE + SF + TE + OE). In vitro 24 h fermentations with varying inclusion levels (0, 0.75, 1.5, 3% of the substrate DM), in comparison with the CON, showed that the T2 group significantly outperformed the other four formulations in nitrogen metabolism and IVDMD, while substantially reducing methane content. Simpson’s index of microbial diversity was markedly elevated, alongside an increased relative abundance of key bacterial genera—Selenomonas and Succinibacter. Considering comprehensive performance metrics and economic viability, the 1.5% T2 was selected as the optimal formulation.