The present study investigated the prevalence of uropathogens and antimicrobial potential of three ayurvedic herbs viz., anantmul (Hemidesmusindicus), gulkhair (Malvasylvestris) and manjishtha (Rubiacordifolia) against the urinary tract infection pathogens.E. coliwas the most prevalent pathogen (42%) while Alcaligeneswas least prevalent (3%). The methanolic extracts were found to more effective as compared to aqueous extracts. The methanolic extract of anantmul exhibited maximum antibacterial property against E. coli(18.3±0.47mm) while aqueous extract of exhibited maximum effect upon Alcaligenes(13.7±0.47 mm). The methanolic extract of gulkhair showed highest potential against E. coli (23.7±0.94 mm) while aqueous extract of gulkhair exhibited maximum antimicrobial effect against E. coli (18.7±0.25 mm). The methanolic extract of manjishtha exhibited maximum antibacterial property against E. coli(29.3±0.47mm)while aqueous extract was most effective against E. coli (18.0±0.47 mm).The ayurvedic herbs exhibited significant antimicrobial potential.