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目的参考联合国粮农组织/世界卫生组织(FAO/WHO)食品添加剂联合专家委员会推荐的方法,在对包括纸片规格、培养基、标准菌株浓度、阳性对照物浓度等影响酶制剂抗菌活性检测主要因素优化的基础上,建立微生物源酶制剂抗菌活性检测方法。方法以浓度为5.0和10.0μg/片的环丙沙星纸片为阳性对照,在相同的培养温度、培养时间等试验条件下,比较9种不同规格纸片、3种品牌培养基、3个菌液稀释度对6种标准菌株抗菌活性的影响。结果 HZT 2号滤纸、2张叠加的whatman903纸片与商业化抗菌活性测定用纸片Whatman 2017-013效果相当;国产1号和2号胰蛋白胨大豆琼脂(TSA)与进口TSA效果相当;确定5种受试菌培养24 h后菌液的10倍稀释液为工作浓度(化脓链球菌的工作浓度为菌液的20倍稀释液);环丙沙星纸片的工作浓度为5.0μg/片。结论HZT 2号滤纸、2张叠加的Whatman903滤纸或Whatman 2017-013滤纸、国产或进口TSA培养基、受试微生物培养24 h的10倍(化脓链球菌20倍)菌液稀释液、环丙沙星浓度5.0μg/片为微生物源酶制剂抗菌活性测定中优化出的最佳试验条件。
OBJECTIVE To determine the main factors affecting the antibacterial activity of enzyme preparations in reference to the methods recommended by the Joint FAO / WHO Expert Committee on Food Additives (JECFA), including the specifications of paper, medium, concentration of standard strains, concentration of positive control, Optimization, based on the establishment of microbial source enzyme preparation antibacterial activity detection method. Methods Ciprofloxacin (5.0 and 10.0 μg / tablet) was used as a positive control. Under the same culture temperature and culture time, 9 different types of media, 3 media Effect of Bacterial Dilution on the Antibacterial Activities of 6 Standard Strains. Results HZT filter paper No. 2 and two sheets of whatman903 paper were equivalent to those of Whatman 2017-013, a commercially available antibacterial activity test paper. The results showed that domestic TSA No. 1 and No. 2 were equivalent to imported TSA. The tested bacteria cultured for 24 h after the 10-fold dilution of the working fluid concentration (Streptococcus pyogenes working concentration of 20 times the dilution of the bacterial liquid); ciprofloxacin paper working concentration 5.0μg / piece. Conclusion HZT filter paper No. 2, two Whatman903 filter paper or Whatman 2017-013 filter paper, domestic or imported TSA medium, 10 times (20 times of Streptococcus pyogenes) bacterial culture of the test microorganism cultured for 24 hours, Star concentration of 5.0μg / tablet for the microbial source of antibacterial activity of the enzyme activity optimization of the optimal test conditions.