Shotgun metagenomic sequencing of ruminal microbial species in beef steers supplemented with increasing doses of thymol

Four beef steers were utilized in a 4x4 Latin square experiment that included four 14-d periods. Steers consumed a basal forage diet ad libitum and received one of four doses of thymol (0 [CON], 120 [120-T], 240 [240-T], and 480 [480-T] mg thymol/kg intake) administered on alfalfa cubes. Rumen contents were collected then separated into liquid and solid fractions on d 14 of each period. Microbial DNA was extracted from liquid and solid rumen contents for subsequent shotgun sequencing.