The objective of this study was to evaluate whether different severities of short-term feed restriction (FR) affect the absorptive function of the reticulo-rumen and total tract barrier function in beef cattle. Eighteen ruminally cannulated and ovariectomized Angus × Hereford heifers were blocked by BW into 3 blocks, with blocks conducted sequentially. Treatments were imposed during the 5-d FR period by restricting heifers to 75 (FR75), 50 (FR50) or 25% (FR25) of the ad libitum feed intake measured during a 5-d baseline period (BASE) occurring immediately before FR. Throughout the study, heifers were housed in individual pens (9 m(2)) and were fed the same diet (60% forage:40% concentrate) with free access to water. Dry matter intake was measured daily and ruminal pH was measured every 2 min throughout the study. Ruminal fluid and blood samples were collected on d 3 of the BASE and FR periods, and the temporarily isolated and washed reticulo-rumen technique was used to evaluate short-chain fatty acid (SCFA) absorption on d 5 of the BASE and FR periods. Total tract barrier function was evaluated starting on d 2 of the BASE and FR periods using a pulse dose of Cr-EDTA followed by 48 h of total urine collection. Data were analyzed using the Proc Mixed procedure of SAS with the fixed effects of block, treatment, period, and the treatment × period interaction, the random effect of cow nested in block with period included as a repeated measure. Dry matter intake did not differ among treatments during BASE but, as imposed by the experimental model, DMI during FR relative to BASE equated to 70, 49, and 25%, which was close to the targeted values of 75, 50, and 25% (treatment × period, P < 0.001). A treatment × period interaction (P < 0.001) was also detected for ruminal SCFA concentration with the concentration decreasing as the severity of FR increased, whereas there were no differences during BASE. Mean ruminal pH increased during FR with increasing severity of FR, but was not different during BASE (treatment × period, P < 0.001). Absorption of SCFA across the reticulo-rumen tended to decrease with increasing severity of FR (P = 0.08). For individual SCFA, acetate absorption (mmol/h) tended (P = 0.057) to be less for FR25 and FR50 when compared with FR75 and decreased (P = 0.05) by almost 70 mmol/h at FR25 and FR50 relative to BASE (322mmol/h). Heifers restricted to 25% (FR25) feed had greater urinary Cr recovery during FR than BASE, whereas no changes were detected for FR75 and FR50. This study indicates that moderate severities of short-term FR decrease the absorptive function of the reticulo-rumen, but more severe FR is required to compromise total tract barrier function in beef cattle.