In Vitro Antibacterial Activity Of The Coelomic Fluid And Deoxyribonuclease Of The Eisenia Foetida Earthworm

ObjectiveTo detect the antibacterial activity in the coelomic fluid of the earthworm and determine its minimal inhibitory concentration and minimum bactericidal concentration. To study the antibacterial mechanisms of the coelomic fluid and deoxyribonuclease of the Eisenia foetida earthworm.MethodsThe extract of the coelomic fluid of the earthworm was prepared with a series of steps, such as treatment in ammonium carbonate, acetone precipitation. the coelomic fluid of the earthworm was determined minimal inhibitory concentration and minimum bactericidal concentration by double dilution method.The effect of coelomic fluid on the integrity of outer membrane of E. coli was examined by NPN uptake factor. The coelomic fluid of the earthworm was further tested against the peptidoglycan from Gram-positive bacteria. The effect of coelomic fluid on the cytoplasmic of E. coli was examined by ONP OD450nm value. CATB was used to extract the bacterial genomic DNA with and alkaline lysis method was used to extract the bacterial plasmid DNA. The degradation of bacterial DNA by the coelomic fluid was observed with agarose gel electrophoresis.Biofilm formation assay and quantification was used for the evaluation of DNase antibiofilm activity. Combined action of the coelomic fluid of the earthworm and deoxyribonuclease on bacteria was tested in order to know the antibacterial mechanisms of deoxyribonuclease of the Eisenia foetida earthworm.ResultsThe coelomic fluid of the earthworm has antibacterial activity. The antibacterial activity of bacteria-induced coelomic fluid was higher than the antibacterial activity of non bacteria-induced coelomic fluid. The coelomic fluid of the earthworm has permeabilization against the cytoplasmic and outer membrane and can degrade peptidoglycan, plasmid DNA and bacterial genomic DNA. Earthworm DNase inhibit the biofilm formation of bacteria.ConclusionThe coelomic fluid of the earthworm can kill bacteria by penetrating the cytoplasmic and outer membrane, degrading peptidoglycan, plasmid DNA and bacterial genomic DNA. The coelomic fluid of the earthworm has selectivity to bacteria.Earthworm DNase participate in innate immunity.