In the present study we established a method for the determination of faecal glucocorticoid metabolites in dogs and then used the assay to evaluate the adrenocortical activity in 12 dogs divided into two groups. In group A faecal samples were collected at their domestic setting. In group B, faecal samples were collected at home prior to transport to a boarding kennel, where faecal samples were then collected. In faecal samples most of the steroids were extracted with methanol and determined using a radioimmunoassay with an anti-cortisol antibody. Dogs in group A did not show any statistically significant inter-day variations in the basal levels of faecal corticoid metabolites. Faecal corticoid metabolites in dogs in group B were significantly higher on the first day at the kennel compared to animals kept at home. The peak concentration was found after 24 hours and followed by a slow decline. These results suggest that extraction with methanol and dosage with an anti-cortisol antibody by radioimmunoassay represents a valid approach technique for determination of faecal glucocorticoid metabolites and accurately reflects adrenocortical activity.