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Intestinal barrier function in pigs after weaning is almost exclusively determined in terminal experiments with Ussing chambers. Alternatively, the recovery in urine of orally administered lactulose can be used to assess intestinal permeability in living animals. This experiment was designed to study the barrier function of the small intestine of pigs over time after weaning. The aim was to relate paracellular barrier function (measured by lactulose recovery in the urine) with macromolecular transport [measured by horseradish peroxidase (HRP) using Ussing chambers] and bacterial translocation to assess whether lactulose recovery is related to possible causes of infection and disease. Forty gonadectomized male pigs (6.7 ± 0.6 kg) were weaned (d 0) at a mean age of 19 d, fitted with urine collection bags, and individually housed. Pigs were dosed by oral gavage with a marker solution containing lactulose (disaccharide) and the monosaccharides L-rhamnose, 3-O-methylglucose, and D-xylose at 2 h and at 4, 8, and 12 d after weaning. The recovery of sugars in the urine was determined over 18 h after each oral gavage. The day after each permeability test, the intestines of 10 pigs were dissected to determine bacterial translocation to the mesenteric lymph nodes and jejunal permeability for HRP in Ussing chambers. Recovery of L-rhamnose in urine was affected by feed intake and by the time after weaning (P≤ 0.05). Recovery of lactulose from the urine was greater (P≤ 0.05) at 4, 8, and 12 d after weaning compared with the first day after weaning and was negatively correlated with feed intake (r = −0.63, P≤ 0.001). The mean translocation of aerobic bacteria to the mesenteric lymph nodes was greater at 5 and 13 d after weaning compared with d 1 (P≤ 0.05). Lactulose recovery showed no correlation with permeability for HRP nor with bacterial translocation (P> 0.05). Although both lactulose recovery and bacterial translocation increased over time after weaning, lactulose recovery did not correlate with the permeability for HRP nor bacterial translocation within a pig (P> 0.05). Therefore, we conclude that lactulose recovery in the urine of pigs after weaning is not associated with risk factors for infections. However, it appears to be possible to measure paracellular barrier function with orally administered lactulose in pigs shortly after weaning. Further studies will reveal whether this variable is relevant for the long-term performance or health of pigs after weaning.