In order to solve problems that number of viable cells in piglet paratyphoid live vaccine declined during storage and transportation, the experiment applied different protective matrix to constitute lyoprotectant, mixed with piglet paratyphoid Salmonella, to prepare piglet paratyphoid heat-resistant protective agent live vaccine after lyophilization. The lyophilization protection rate and heat-resistant protection rate of the live vaccine are determined, to evaluate quality of vaccine. The results show that: T1 and T2 are preferred piglet paratyphoid heat-resistant protection agents, in three batches of piglet paratyphoid heat-resistant protective agent live vaccine trially produced by using T2 protective agent, after storing at 37°C for 10 days, the heat-resistant protective agent of vaccine is more than 80%, and greater than 3 × 109 cfu/parts. The live vaccine has good thermal stability, is safe and effective in field trials.