Antimicrobial activity of a Ruthenium (III) complex with N-phenyl-5-chlorosalicylideneimine ligands have been studied against gram-positive and gram-negative pathogenic bacterial strains by using disc diffusion method. The compound showed significant in vitro antimicrobial activities against community acquired methicillin-resistant Staphylococcus aureus, hospital acquired methicillin-resistant Staphylococcus aureus, methicillin-sensitive Staphylococcus aureus, Bacillus cereus ATCC 11778, Bacillus subtilis ATCC 6633 and Enterococcus faecalis ATCC 29212. The minimal inhibitory concentration values of the compound were found to be in the range of 11.72–23.44 μg/mL. Additionally, minimal bactericidal concentration values and considerable sensitivity pathogenic gram–positive microorganisms on Ru(III) compound against led to conclude the complex have prospective antibiotic properties. The compound has no activity in vitro against gram-negative bacteria Klebsiella pneumonia, Enterobacter species, Pseudomonas aeruginosa, Escherichia coli ATCC 25922 and Salmonella typhimurium ATCC 14028.