For the selective isolation and enumeration of Staphylococci. MANNITOL SALT AGAR is used for the selective isolation and enumeration of Staphylococci species. It conforms to harmonized USP/EP/JP requirements.1,2,3 Chapman formulated Mannitol Salt Agar to isolate staphylococci by inhibiting growth of most other bacteria with a high salt concentration.4Mannitol is the fermentable carbohydrate which leads to acid production, detected by Phenol red indicator. Staphylococcus aureus ferment mannitol and produce yellow coloured colonies surrounded by yellow zones. The lipase activity can be visualized as yellow opaque zones around the colonies.5 Coagulase-negative strains of Staphylococcus aureus are usually mannitol non-fermenters and therefore produce pink to red colonies surrounded by red-purple zones. Presumptive coagulase-positive yellow colonies of Staphylococcus aureus should be confirmed by performing the coagulase test. Lipase activity of Staphylococcus aureus can be detected by supplementing the medium with egg yolk emulsion. Sodium chloride in high concentration serves as an inhibitory agent against bacteria other than Staphylococci. Beef extract gives essential growth factors and trace nutrients to the growing bacteria. Phenol Red is the pH indicator. Agar acts as the solidifying agent. Inoculate Mannitol Salt Agar (TMH 114) medium with a small number (not more than 100 cfu) of the appropriate microorganism and incubate at 35 - 37°C for 18 - 48 hours. Recovery rate is considered as 100% by sub culturing on Soyabean Casein Digest Agar (TMH 103) at 35±2°C for 18 - 48 hours.