Abstract: Plants produce a diverse range of bioactive molecules, making them rich source of different types of medicines. In the present study, the in vitro antimicrobial activity of medicinal plant Parthenium hysterophorus, was evaluated by using chloroform, methanol, acetone, ethyl acetate, petroleum ether and distilled water. The extracts were tested against Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Bacillus subtilis, Saccharomyces cerevisiae and Staphylococcus aureus by using agar well diffusion method. Ciprofloxacin and Amphotericin were used as standard antibiotics. Some of the solvent extracts of the plant showed the highest activity against some pathogenic microorganisms than standard antibiotics used. The findings provide support for the use of this plant in producing new bioactivity compounds having antimicrobial activity.