To check the antimicrobial activity of Tinospora cardifolia (Giloy) growing on host plants Azadirachta indica (Neem), Saraca asoca (Ashoka), Citrus limon (lemon), on common endodontic pathogens like Escherichia coli, Salmonella, Staphylococcus aureus, Streptomyces, Enterococcus faecalis, and Listeria. The agar diffusion test was used to check the antimicrobial activity of the Methanolic extracts of the medicinal plants with different concentrations. The values of the Zone of Inhibition were tabulated according to the concentration. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values were also recorded. Extracts of all the Tinospora cordifolia growing on host plants showed considerable antimicrobial activity against selected pathogens. At a minimum 50mg/ml conc. all hosted and normal giloy shows zone of inhibition like- against E. coli.- Saraca asoca, against Enterococcus- Citrus limon (Azadirachta indica also), against Listeria- Azadirachta indica, against Streptomyces -Azadirachta indica, against Salmonella- Azadirachta indica (Saraca asoca), and S.aureus- Azadirachta indica (Saraca asoca and Citrus limon also, but less than (Azadirachta indica) shows maximum zone of inhibition. The Methanolic extract of Tinospora cardifolia (Giloy) grows on host plants Azadirachta indica (Neem), Saraca asoca (Ashoka), Citrus limon (lemon), and normal giloy has considerable antimicrobial activity against E. coli, Salmonella, Staphylococcus aureus, Streptomyces, Enterococcus faecalis, and Listeria.