A new series of piperazine-1, 3, 4-thiadiazole has been synthesized, purified and characterized with the help of their analytical and spectral data. Structures of the synthesized compounds were confirmed by IR, 1H NMR and mass spectroscopy. The potential antimicrobial effects of the synthesized compounds were investigated using Vibrio cholera and Bacillus subtilis. The newly synthesized compounds exhibited capable activities against Vibrio cholera and Bacillus subtilis and it showed minimum inhibitory concentration. In this study, few compounds showed appreciable antibacterial activity. The compound PT6 shows highly significant antibacterial and good inhibition as compared to the standard drug.