Cobalt (Co) doped zinc oxide (ZnO) microcrystals (MCs) are prepared by using the hydrothermal method from the precursor's mixture of zinc chloride (ZnCl2), cobalt-II chloride hexahydrate (CoCl2·6H2O), and potassium hydroxide (KOH). The smooth round cylindrical morphologies of the synthesized microcrystals of Co-doped ZnO show an increase in absorption with the cobalt doping. The antibacterial activity of the as-obtained Co-doped ZnO-MCs was tested against the bacterial strains of gram-negative (Escherichia coli, Klebsiella pneumonia) and gram-positive bacteria (Staphylococcus aureus, Streptococcus pyogenes) via the agar well diffusion method. The zones of inhibition (ZOI) for Co-doped ZnO-MCs against E. coli and K. pneumoniae were found to be 17 and 19 mm, and 15 and 16 mm against S. Aureus and S. pyogenes, respectively. The prepared Co-doped ZnO-MCs were thus established as a probable antibacterial agent against gram-negative bacterial strains.