A safety method for rhinogobio ventralis long-distance transportation comprises the following steps of 1 controlling the temperature of a water body; 2 adding penicillin; 3 adding anesthetic to perform first-step anesthesia; 4 rapidly transferring rhinogobio ventralis to a transportation water body of a transportation device; 5 adding the anesthetic to perform second-step anesthesia; 6 adding oxygen into the transportation water body, namely ensuring 10-20hour long-distance transportation safety of the rhinogobio ventralis. A safety transportation device for rhinogobio ventralis long-distance transportation comprises a box body and a box cover which are connected through hinges, the box body consists of an inner vat, an ice storage vat and a heat preservation layer which are sequentially arrange from inside to outside, and an ice storage layer is formed between the inner vat and the ice storage vat. The box cover is provided with a groove which is in a shape of a Chinese character 'hui' and matched with the inner vat and the ice storage vat. The safety method and the transportation device for the rhinogobio ventralis long-distance transportation greatly improve the survival rate of the rhinogobio ventralis in the long-distance transportation and achieve the 10-20hour long-distance transportation safety.