Chocolate agar


Image:Yersinia_pestis_HHS.jpg|thumb|250px|Colonies of Yersinia pestis growing on chocolate agar
Image:Chocolate agar 1.jpg|thumb|250px|Chocolate agar showing Francisella tularensis colonies
Image:Neisseria gonorrhoeae 01.png|thumb|300px|Comparison of two culture media types used to grow Neisseria gonorrhoeae bacteria.

Known as overgrowth, the nonselective chocolate agar medium on the left, due to its composition, allowed for the growth of organismal colonies other than those of N. gonorrhoeae, while the selective Thayer–Martin medium on the right, containing antimicrobials that inhibit the growth of organisms other than N. gonorrhoeae, shows no overgrowth, but is positive for N. gonorrhoeae bacteria.
Chocolate agar or chocolate blood agar is a nonselective, enriched growth medium used for isolation of pathogenic bacteria. It is a variant of the blood agar plate, containing red blood cells that have been lysed by slowly heating to 80°C. Chocolate agar is used for growing fastidious respiratory bacteria, such as Haemophilus influenzae and Neisseria meningitidis. In addition, some of these bacteria, most notably H. influenzae, need growth factors such as nicotinamide adenine dinucleotide and hemin, which are inside red blood cells; thus, a prerequisite to growth for these bacteria is the presence of red blood cell lysates. The heat also inactivates enzymes which could otherwise degrade NAD. The agar is named for its color and contains no chocolate products.

Variants

Chocolate agar with the addition of bacitracin becomes selective for the genus Haemophilus. Another variant of chocolate agar called Thayer–Martin agar contains an assortment of antibiotics that select for Neisseria species.

Composition

The composition of chocolate agar includes the following components:
IngredientQuantity Purpose
Casein/Animal Tissue Digest15.0 gramsSource of amino acids and peptides for bacterial growth.
Cornstarch1.0 gramsProvides carbohydrates as an energy source for bacteria.
Sodium Chloride5.0 gramsMaintains osmotic balance in the medium.
Dipotassium Phosphate4.0 gramsActs as a pH buffer to maintain the desired pH for bacterial growth.
Monopotassium Phosphate1.0 gramsActs as a pH buffer to maintain the desired pH for bacterial growth.
Hemoglobin Solution 2%500 mlProvides iron and other nutrients necessary for bacterial growth.
Coenzyme Enrichment10 mlContains coenzymes needed for specific metabolic pathways of bacteria.
Agar10 gramsSolidifying agent that allows the medium to form a solid surface for bacterial growth.

The exact concentrations of these ingredients may vary slightly depending on the specific formulation used in different laboratories or by different manufacturers.

History

Addition of heated blood to media was first documented for use by Cohen and Fitzgerald in 1910 and then by Dr. Olga Povitzky at the New York City Department of Health Bureau of Laboratories. The term "chocolate" comes from the brown color generated from the higher concentration of heated blood in the mixture and was first used by Warren Crowe in 1915.