Columbia CNA (Colistin-nalidixic acid) agar

Intended use: Columbia CNA Agar with 5% Sheep Blood is a selective and
differential medium for the isolation and differentiation of gram positive
microorganisms from clinical and nonclinical specimens.
Chief ingredients: Per Liter Purified Water
Pancreatic Digest of Casein
Sodium Chloride
Peptic Digest of Animal Tissue
Agar
Yeast Extract
Colistin (Polymyxin B)
beef Extract
Nalidixic Acid
Corn Starch
Sheep Blood, defibrinated

12.0g
5.0g
5.0g
13.5g
3.0g
10.0 mg
3.0g
10.0 mg
1.0g
5%

Preparation:
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6.

Suspend 44.0g of the medium in 1000ml distilled water

Boil to dissolve
Sterilize by autoclaving at 121 C for 15 mins
Cool to 50 C
Add 5% v/v sterile defibrinated horse or sheep blood as required
Dispense into appropriate containers, allow to set

Principles: This agar has superior growth-supporting properties from the

combination of peptones prepared from pancreatic digest of casein and yeast
extract.
Yeast extract and corn starch are included in the formulation and serve as energy
sources. 5% Sheep blood allows detection of hemolytic reactions and supplies the X
factor (hemin) necessary for the growth of many bacterial species but lacks V factor
(nicotinamide adenine dinucleotide, NAD).
The addition of the antimicrobial agents, colistin and nalidixic acid, renders the
medium selective for gram-positive microorganisms. The colistin disrupts the cell
membranes of gram-negative organisms, whereas the nalidixic acid blocks DNA
replication in susceptible gram-negative bacteria.
It should be noted that this medium has a relatively high carbohydrate content and,
therefore, beta-hemolytic streptococci may produce a greenish hemolytic reaction
that may be mistaken for alpha hemolysis.

Reference: The manual of microbiological culture media, Life Technology,

http://www.lifetechnologies.com/nz/en/home/life-science/cell-culture/microbiologicalculture.html