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Skin barrier restoration is an important part of atopic dermatitis therapy. We investigated the effect of a spot-on containing plant-based essential fatty acids and essential oils on skin barrier parameters in a dog model of acute skin barrier disruption, using five healthy beagle dogs maintained in a laboratory setting. Four test sites on the dorsum and a control site on the abdomen were defined on each dog. Transepidermal water loss (TEWL) and skin surface hydration (SSH) were measured before and after tape stripping on the first day and then for three consecutive days, over four consecutive weeks. The spot-on was applied at the end of each of the first three weeks. The increase in TEWL after tape stripping was reduced after the spot-on application and reached control values in Weeks 3 and 4. SSH after tape stripping was reduced in Week 4 compared with the baseline. Thus, the ATOP 7^® spot-on significantly reduced acute skin barrier impairment in a dog model. The use of this product should be further evaluated as a potential treatment for skin barrier defects such as canine atopic dermatitis.