蚯蚓体腔液及粗组分体外抗菌特性

In Vitro Antibacterial Effect of Earthworm Ceolomic Fluid and Crude Extracts

利用钢圈法测定蚯蚓体腔液对大肠杆菌(Escherichiacoli)、绿脓杆菌(pseu-domonasaeruginosa)和枯草芽孢杆菌(Bacillussubtilis)的抑制作用,并利用微量平板稀释法测定蚯蚓体腔液对绿浓杆菌的最低抑菌浓度。结果如下:蚯蚓体腔液对枯草芽孢杆菌和大肠杆菌没有抑制作用、对绿脓杆菌有抑制作用,浓度为0.1g/ml时绿脓杆菌的抑菌圈直径为1.53cm;蚯蚓体腔液对绿脓杆菌的最低抑菌浓度(MIC)为6.25mg/mL。体腔液用85%的硫酸铵沉淀、截留分子量为10kDa和1kDa的超滤膜进行超滤,得到分子量分别为大于10kDa、1kDa～10kDa、小于1kDa的三种组分ECP1、ECP2、ECP3。用钢圈法测定了ECP1、ECP2、ECP3对绿脓杆菌的抑制作用,其中ECP3的抑菌作用最强,ECP1、ECP2和ECP3的抑菌圈直径分别为7.68mm、11.44mm和16.8mm。ECP3对绿脓杆菌的最低抑菌浓度为0.018mg/mL。

The antibacterial effect of coelomic fluid to Escherichia coli, pseudomonas aeruginosa, Bacillus subtilis, and the MIC of coelomic fluid to pseudomonas aeruginosa was studied with steel ring method, and standard micro-dilution method respectively. The results showed that the coelomic fluid had a great inhibiting effect on pseudomonas aeruginosa, with a 1.53 cm diameter inhibiting zone and had no effect on Escherichia coli, and Bacillus subtilis. The MIC of coelomic fluid to pseudomonas aeruginosa was 6.25 mg/mL. The coelomic fluid was precipitated with ammonium sulfate at a final concentration of 85% (w/v), then was ultrafiltrated by 1 kDa and 10 kDa molecular weight cut-off (Shanghai research institute) ultrafitration membranes, at last three components named ECP1, ECP2, and ECP3, with a molecular mass of 10 kDa, 1 kDa^10 kDa, <1 kDa respectively were abtained. The antibacterial effect of ECP1, ECP2, ECP3 to pseudomonas aeruginosa and the MIC of ECP3 to pseudomonas aeruginosa was studied with steel ring method and standard micro-dilution method respectively. The diameter of the inhibiting zone of ECP3 was 16.8 mm, ECP1, ECP2 were 7.68 mm, 11.44 mm respectively. The MIC of ECP3 to pseudomonas aeruginosa was 0.018 mg/mL.