<b>A Comparative in Vitro Antimicrobial Activity of Annona Squamosa on Gram Positive and Gram Negative Microorganism</b> Annona squamosa L is a multipurpose tree with edible fruits and is a source of the medicinal and industrial products. It is used as an antioxidant, antidiabetics, hepatoprotective, cytotoxic, genetoxic, anti tumor, anti lice agent etc.Annona squamosa L belongs to the family Annonaceae commonly known as custard apple. Antimicrobial activity of combined methanolic leaf and seed extract of A.squamosa were evaluated against four bacteria Bacillus subtilis, Styphaloccocus aureus gram positive and E.coli, Pseudomonas aerogenosa by using cup and plate method. Maximum inhibition was found with 20mg ml concentration of combined extract as compare to separate leaf and seed extract against all the tested organism under investigation. The study suggest that maximum antibacterial activity was observed against gram negative organism i.e., E.coli and P.aerogenosa. Annona squamosa L, antimicrobial activity, cup and plate method. 2492-2496 Issue-5 Volume-3 Ms. Chetana D. Patil | Ms. Nikita Pawar | Mrs. Pooja S. Bhandare