Envisaging the role of Co in theranautics and biomedicine it is immensely important to evaluate its antimicrobial activity. Hence in this study CoO thin nanosheets (CoO-TNs) were synthesized using wet chemical solution method at a very low refluxing temperature (90 oC) and short time (60 min). Scanning electron microscopy of the grown structure revealed microflowers (2~3 μm) composed of thin sheets petals (60~80 nm). The thickness of each individual grown sheet varies from 10~20 nm. Antimicrobial activities of CoO-TNs against two Gram positive bacteria (Micrococcus luteus, and Staphylococcus aureus), and two Gram negative bacteria (Escherichia coli and Pseudomonas aeruginosa) were determined. A 98% and 65% growth inhibition of M. luteus and S. aureus respectively, was observed with 500 μg/ml of CoO-TNs compared to 39 and 34% growth inhibition of E. coli and P. aeruginosa, respectively with the same concentration of CoO-TNs. Hence, synthesized CoO-TNs exhibited antimicrobial activity against Gram negative bacteria and an invariably higher activity against tested Gram positive bacteria. Therefore, synthesized CoO-TNs are less prone to microbial infections.