[Objectives] The demand for ponies has been increasing in recent years, but the stock of Debao ponies is far from enough to supply market demand. In this context, this paper aims to achieve the purpose of conservation, purification and rejuvenation. [Methods] Under the conditions of same age, semen collection frequency, season, feeding environment, feed nutrition level, etc., the semen of Debao pony and American pearl pony were collected, respectively, and the quality of the semen collected was analyzed. [Results] There was no significant difference in semen volume between the two breeds of ponies; the sperm vitality of the firstly-collected semen of ponies of which the semen has not been collected for a long time was low. The semen quality of American pearl pony was slightly better than that of Debao pony. [Conclusions] When the semen quality of American pearl pony is stable, it can be used for cross-breeding with the Debao pony mare, highlighting the hybrid vigor of the two breeds, thereby achieving the purpose of reducing the size of ponies.