Objective To establish a stable and efficient asthma model in beagle dogs. Methods The antigen of ragweed short (Ambrosia elatia, RS) pollen or mixed antigens of both house dust mite (Dermatophagoides farinae, HDM) and RS pollen were prepared. Healthy beagle dogs were sensitized by injecting mixed antigens within 24 hours after birth and at 1, 4, and 8 weeks. From week 1 to 12, RS pollen antigen or mixed antigens were inhaled every day (daily nebulization group, DN) or twice a week (intermittent nebulization group, IN) for airway challenge. From week 13, RS pollen antigen was inhaled once a week until week 36. At 36 and 48 weeks, an airway responsiveness test, differential counting of leukocytes in alveolar lavage fluid, and pathological analysis were carried out to evaluate the outcomes of asthma modeling. Results At 36 weeks, neutrophilic and eosinophilic inflammation in airways and airway hyperresponsiveness were observed in DN and IN groups. At 48 weeks, airway eosinophilic inflammation in DN and IN groups, and airway hyperresponsiveness in the DN group persisted. Asthma-related airway remodeling was observed in both groups. Conclusions An asthma model in beagle dogs can be established by sensitizing and challenging newborn dogs with mixed antigens of RS pollen and HDM, and increasing the frequency of antigen inhalation challenge.