Traditionally, Cynodon dactylon L. juice is used for freshness and several common diseases. Preliminary phytochemical analysis proved that phenols, quinines and tannin were in ethanolic extract of C. dactylon. Then the ethanolic extract was analyzed by Gas Chromatography and Mass Spectrophometry (GC-MS) which showed 10 phytochemical components, some of them are not mentioned in the previous studies among these Tricosane (22.05 %), 1, 2-Propanediol (20.30%), 3-benzyloxy-1, 2-diacetyl (12.62%) were present at maximum level. Aqueous and ethanolic extract of C. dactylon (500 µg/ml) were investigated for their antibacterial activity against gram positive bacteria and gram negative bacteria using disc diffusion, well in agar and microdilution method. E. coli, B. subtilis, S. aureus and A. hydrophila were more susceptible in the ethanolic extract and no result was found in aqueous extract. Minimum inhibitory concentrations (MIC) value of the ethanolic extract was response between in the range of 125 - 62.5 µg/ml. Due to the presence of phyto components may control the bacterial growth which supports the usage of plant extract either in high concentration/long duration of traditional treatment by the traditional healers.