Abstract: Clostridium perfringens（Cp）is the leading pathogen of necrotizing enteritis——an acute zoonotic disease. Reuterin is a product of few probiotics in glycerol substrate，which exhibited broad spectrum antibacterial activity. The objective of this study was to observe the growth inhibition feature of Reuterin in vitro on different types of C. perfringens and to explore the stability of Reuterin under different storage temperatures. Lactobacillus reuteri was induced to synthesize Reuterin from glycerol，its concentration was determined by Acrolein ?Tryptophan method. The minimum inhibitory concentration（MIC）of C. perfringens types A，B，C，D and E were determined by 96?well plate OD600 measurement and plate titration. The antibacterial properties of Reuterin against C.perfringens was compared after 24 hours storage at - 20 ℃ ，room temperature（25 ℃）and 37 ℃ . The results exhibited that the Reuterin production concentration was 26.13 μmol/mL. Type A and D C. perfringens were the most sensitive to the Reuterin， with MIC values of 0.78 μmol/mL and 0.52 μmol/mL，respectively. Type B，C and E strains were also sensitive，MIC values were 1.57 μmol/mL，1.57 μmol/mL and 1.83 μmol/mL. In the plate titration method，all five serotype bacteria were inhibited under 0.26 μmol/mL Reuterin treatment. The bacteriostatic effect of Reuterin were measured after different storage temperatures. For C.perfringens type A and C，the growth inhibition capacity did not change，and the MICs were 0.78 μmol/mL and 1.57 μmol/mL. This study proved that Reuterin has the grow inhibitory ability on C. perfringens in vitro as one of the probiotic metabolits，and its antibacterial ability is not affected by storage temperature.

Key words: Reuterin； Clostridium perfringens； Minimum inhibitory concentration（MIC）； Temperature stability

摘要： 产气荚膜梭菌（Clostridium perfringens，Cp）是一种急性人畜共患病——坏死性肠炎的 主要病原菌。罗伊氏菌素（Reuterin）是少数益生菌以甘油为发酵底物的产物，具有广谱抗菌活性。 本研究旨在观察罗伊氏菌素对产气荚膜梭菌不同细菌型的体外抑制作用，以及不同储存温度对 罗伊氏菌素抑菌性能的影响。利用甘油诱导罗伊氏乳杆菌合成罗伊氏菌素，通过丙烯醛?色氨酸 法测定其浓度。以96孔板OD检测法和平板滴定法测定罗伊氏菌素对A型、B型、C型、D型和E型 5种产气荚膜梭菌标准菌株的最小抑菌浓度（minimum inhibitory concentration，MIC）。比较-20 ℃、 室温RT（25 ℃）及37 ℃储存24 h后的罗伊氏菌素对产气荚膜梭菌A型和C型的抑菌性能。结果表 明：罗伊氏菌素样品浓度为26.13 μmol/mL。不同浓度罗伊氏菌素处理后，96孔板法抑菌性能检 测中A型和D型最为敏感，MIC值分别为0.78 μmol/mL和0.52 μmol/mL，B型、C型和E型菌株也 较敏感，MIC值分别为1.57 μmol/mL，1.57 μmol/mL和1.83 μmol/mL；平板滴定法中0.26 μmol/mL 罗伊氏菌素处理后5种标准菌已全部被抑制。罗伊氏菌素经不同储存温度处理后测定抑菌性能， 以产气荚膜梭菌 A 型和 C 型为例，结果抑菌性能未发生变化，其 MIC 分别为 0.78 μmol/mL 和 1.57 μmol/mL。本研究证明，罗伊氏菌素对产气荚膜梭菌具有较好的体外抑制作用，且其抑菌性 能不受储存温度影响。

关键词: 罗伊氏菌素； 产气荚膜梭菌； 最小抑菌浓度； 温度敏感性