Physiological biomarkers of canine anxiety have not been extensively investigated to date. To identify new biomarkers in dogs, we compared behaviorally normal dogs (Control group, N = 13) to dogs diagnosed with separation problems (Case group, N = 13) as they were introduced into a novel environment in the presence of two strangers and subjected to a short episode of separation and reunion with the owner. During the separation phase, dogs in the Case group explored significantly less than controls and were significantly more persistent in expressing passive stress-coping strategies aimed at seeking proximity to their owners. When the owners returned, dogs with separation distress spent significantly more time jumping up on the strangers than control dogs did. Salivary oxytocin and vasopressin concentrations did not differ between samples taken before and after the separation. However, vasopressin concentrations immediately after separation were significantly higher in the Case than in the Control group and remained higher, although not significantly so, 10 min later. These results indicated that dogs with separation distress became more anxious than typical dogs when separated from their owner in an unfamiliar environment and provided preliminary support for the use of salivary vasopressin as a possible biomarker for anxiety-related responses in dogs.
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