The utility model discloses an anaerobic cultivation device of probiotics for feeding, which comprises a first cultivation chamber and a second cultivation chamber, wherein the first cultivation chamber and the second cultivation chamber are communicated with each other, anaerobe meeting one requirement is cultivated in the first cultivation chamber and enters the second cultivation chamber through a second conduit. Anaerobe meeting two requirements is cultivated in the second cultivation chamber. The anaerobic cultivation device of probiotics for the feeding has simple structure and low manufacturing cost, a liquid transferring gun is not required between the two cultivation chambers, and liquid transfer is simple to operate, thereby having low requirement on workers. Simultaneously, connectors between the second conduit and the first cultivation chamber and the second cultivation chamber are all arranged below the upper liquid surface, thereby providing oxygenfree or less oxygen environment for the anaerobe through water seals, effectively reducing the possibility of pollution of the anaerobe and improving quality of the anaerobe.