Exhaled breath condensate hydrogen peroxide concentration, a novel biomarker for assessment of oxidative stress in sheep during heat stress

The present study aimed to measure hydrogen peroxide (H2O2) in exhaled breath condensate (EBC) as an index of oxidative stress induced by heat stress (HS) and to evaluate the antioxidant potential of supranutritional levels of selenium (Se) and vitamin E under HS conditions in sheep. Thirty-two Merino x Poll Dorset ewes (in two replicates) were acclimated to indoor individual pen feeding of a pelleted control diet (0.24 g Se and 10 IU of vitamin E/kg DM) for 1 week. Sheep were then randomly allocated to a 2 x 2 factorial design with the respective factors being dietary Se (0.24 and 1.20 mg/kg DM as Sel-Plex) and vitamin E (10 and 100 IU/kg DM) for 2 weeks. After 1 week acclimation to the diets the sheep were placed in metabolism cages in one of two climatic chambers set to either thermoneutral (18-21 degrees C and 40-50% relative humidity) or HS (28-40 degrees C and 30-40% relative humidity) conditions for 1 week. Average respiration rate and rectal temperature were increased (P < 0.001) during HS. Similarly, H2O2 concentration in EBC and plasma reactive oxygen metabolites were increased significantly (P = 0.041 and P = 0.002, respectively), in sheep exposed to HS. The EBC H2O2 concentration was reduced (P < 0.05) in sheep fed the diet supplemented with supranutritional levels of both Se and vitamin E. Sheep supplemented with antioxidants also had lower respiration rates (191 vs 232 breaths/min, P = 0.012) and rectal temperature (40.33 vs 40.58 degrees C, P = 0.039) under peak HS (1300 hours and 1700 hours) compared with those on the control diet indicating an improved capacity to handle HS. These data suggest that the H2O2 concentration in EBC can be utilised as a novel biomarker to demonstrate oxidative stress induced by HS in sheep.