Intestinal Permeability Assessment in Pigs

Intestinal permeability was assessed in vivo on d 28 and 29 after weaning. Eight pigs per treatment (representative of the median BW of each treatment) were fasted for 2 h and subsequently sedated with a mixture of xylazine (1.5 mg/kg BW) and ketamine (11 mg/kg BW) administered intramuscularly (i.m.) in order to minimize handling stress. After 10 min, animals were intragastrically dosed (gastroduodenal feeding tube, Levin type; VEC Medical) with a marker solution containing 0.15 g mannitol/kg BW (Sigma-Aldrich, Madrid, Spain) and 0.1 g Co-EDTA/kg BW [27 (link)] dissolved in 15 mL deionized water and immediately after were challenged with LPS as previously described. Blood samples were collected by jugular venipuncture 1 h after oral infusion of permeability markers into 5 mL evacuated fluoride/K-oxalate glucose blood collection tubes (BD vacutainer, Madrid, Spain). Plasma mannitol was determined by ultra-high performance liquid-chromatography mass-spectrometry (Xevo G2 TOF, Waters) as previously described [28 (link)]. The plasma concentration of cobalt was determined by atomic absorption spectroscopy [27 (link)].