Botulinum toxin type A (BoNT/A) has been proven to be a safe and effective treatment for wrinkles in human. As well as the anti-wrinkle effects, the anti-pruritic effect of BoNT/A has been revealed from several researches for new therapy. The aim of this study was to investigate the anti-pruritic effect of BoNT/A against histamine-induced pruritus on canine skin. Five clinically healthy beagles were included in the study. All dogs were received 0.05 ml (5 Unit) of BoNT/A on the right dorsal thoracic region as an experiment and the same volume of saline solution was injected on the left dorsal thoracic region as a control, respectively. Intradermal histamine injections were performed four times (before treatment and days 1, 3 and 7 after BoNT/A injection). The severity of pruritus, the diameter and thickness of wheal, the erythema index and cutaneous temperature were assessed. The severity of pruritus was reduced on BoNT/A treated sides, compared with saline treated sides (p < 0.05). BoNT/A decreased wheal size, in both diameter and thickness (p < 0.05). Although, erythema index of both sides were increased after first histamine injection, BoNT/A treated sides showed the low-value as compared with saline treated sides. Cutaneous temperature was decreased significantly on BoNT/A treated sides. These results indicates that BoNT/A reduce histamine-induced pruritus on canine skin and suggested a possibility of application of BoNT/A for local intractable dermatologic problem in dogs.