Types of Media for Microbiology

Blood Agar A rich, non-selective, differential medium which supports the growth of a wide variety of microbes. It is made by adding 5% sheeps blood to a complex medium containing peptones just before the plates are poured. The red blood cells remain intact and the medium resembles fresh blood. Some bacteria grow on blood agar without disturbing the red cells; others hemolyze the cells to various degrees. Bacteria that lyse the red cells with a complete clearing of the medium around the colonies are referred to as beta-hemolytic. Some bacteria lyse the cells but do not break down the hemoglobin completely leaving greenish methemoglobin in the medium. This is called alphahemolysis. Non-hemolytic bacteria have no visible effect on the red blood cells and are said to be gamma-hemolytic. Eosin Methylene Blue Agar (EMB) This agar medium is both selective and differential. The combination of the two dyes eosin and methylene blue inhibits most Gram positive bacteria but allows many Gram negative organisms to grow. In addition to peptones, EMB contains lactose (it may also contain sucrose). Gram negative bacteria that ferment the lactose produce acid which turns the colonies dark purple as the acid acts upon the dyes. In addition, certain lactose-fermenting bacteria produce flat, dark colonies with a green metallic sheen. Other lactose fermenters produce larger, mucoid colonies, often purple only in their center. Lactose non-fermenters are either colorless or light lavender. MacConkey Agar (MAC) This is a differential plating medium, selective for Gram negative organisms, used primarily for detection and isolation of enteric bacteria. Grampositives are generally inhibited by crystal violet in the medium. Isolated colonies of lactose-fermenting bacteria are brick red in color and may be surrounded by a zone of precipitated bile. This reaction is due to the action of the acids, produced by fermentation of lactose, upon bile salts present in the medium, and subsequent absorption of neutral red. Non-lactose fermenters are colorless and transparent. Mannitol Salt Agar (MSA) This is a selective medium that favors the growth of staphylococci and for differentiation between pathogenic and non-pathogenic types. MSA contains 7.5% NaCl and staphylococci are characteristically able to tolerate this high salt concentration. MSA also contains the sugar-alcohol mannitol and the pH indicator phenol red. Generally pathogenic staphylococci are able to ferment the mannitol, lowering the pH, and thereby turning the indicator yellow The non-pathogenic staphylococci do not ferment mannitol and the medium remains pink in their vicinity. Milk Agar This is nutrient agar containing powdered mild. It is used to test for an organisms ability to secrete extracellular proteases that catalyze the hydrolysis of the milk protein casein. A clear zone around a colony indicates casein hydrolysis. Phenylethanol Agar (PEA) Phenylethanol inhibits many Gram negative bacteria and its inclusion in PEA renders the medium selective for Gram positives.

Sabouraud Dextrose Agar (SDA) A medium that promotes the growth of fungi (yeast and molds) but which is inhibitory for many bacteria Simmons Citrate Agar This medium, which may be used in the form of slants or plates, tests for the ability of an organism to grow on citrate as the sole carbon source. It contains various minerals, including ammonium phosphate for nitrogen, sodium citrate, agar, and the pH indicator brom thymol blue. The indicator is green when acid or neutral. As bacteria grow on the medium, they make the medium increasingly alkaline and the indicator turns blue. Spirit Blue Agar This medium is used for detecting lipase activity. It contains nutrients, emulsified lipids and the dye spirit blue. Clearing of the emulsion is a positive indicator of lipase. Some organisms will bring about precipitation of the blue dye. Trypticase Soy Agar (TSA) A general purpose nutrient medium that promotes the growth of bacteria