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Comparison of cortisol, luteinizing hormone, and testosterone responses to a defined stressor in sexually inactive rams and sexually active female-oriented and male-oriented rams¹

USDA-ARS US Sheep Experiment Station, Dubois, ID 83423

² Corresponding author: jstellflug{at}pw.ars.usda.gov

The objective of this study was to determine whether the effect of restraint stress on cortisol, LH, and testosterone varied among sexually inactive and sexually active female- and male-oriented rams, to allow differentiation among ram classes. Restraint stress or no stress was imposed on sexually inactive (n = 7) and sexually active female- (n = 17) and male-oriented (n = 6) rams in a 2 x 3 factorial arrangement. Rams were assigned to restraint or control within each classification. Rams were habituated to wearing halters and being tethered in separate pens, permitting visual, vocal, and olfactory contact with adjacent rams for 7 d before treatment. After 1 d of habituation, rams were fitted with jugular catheters that were checked twice daily for patency. For restraint stress, rams were laid on their side with their legs tied for 1 h. For no stress, rams were tethered with halters and leads, but their legs were not tied. On the treatment day, blood was collected at 30-min intervals for 3 h followed by 15-min intervals for 1 h before restraint, during 1-h restraint, and for 1 h after liberation from restraint. Then blood was collected at 30-min intervals for an additional 2 h. Blood was collected from controls at similar intervals. Control rams were isolated from stressed rams. Cortisol, LH, and testosterone were measured using RIA. Mixed model analyses with repeated measures were used on transformed data. Average prestress data were used as a covariate. Cortisol increased (P < 0.01) within 15 min after restraint and remained increased until 1.5 h after liberation from 1-h of restraint stress. In contrast, in controls cortisol remained unchanged at 5 ng/ mL. Cortisol did not differ over time among ram classes, and the treatment x ram class x time interaction was not significant. For LH and testosterone, the ram class x time interactions appeared to compromise the ability to identify differences in these hormones, indicating that they were not good endocrine candidates for methods of classifying rams. In conclusion, restraint stress increased cortisol in sexually inactive and sexually active female- and male-oriented rams alike, thus not providing a method to differentiate among ram classes.

Key Words: cortisol • luteinizing hormone • ram • sexual performance • stress • testosterone