Dehydrated culture media
Columbia Broth
For the cultivation of a wide variety of fastidious microorganisms or as a general use broth.
- Cat. No.
- LC-1229
- Category
- Dehydrated culture media
- Format
- 500 g
- Storage
- 2 to 25 °C
- Minimum guaranteed shelf life
- 15 months
Columbia Broth, being highly nutritive, is used for the cultivation of microorganisms in general, as even the most fastidious grow in it. This medium is prepared according to the formulation described by Morello and Ellner in their study of Columbia Broth.
Key features
Applications
Growth (general use).
Industry
General cultivation.
Preparation
35 g in one liter of distilled water; boil for one minute, sterilize in autoclave at 121 °C for 15 minutes.
Principles and uses
A medium developed for blood cultures, it is superior to commonly used general media since it allows a faster growth of Staphylococcus aureus, E. coli and streptococci (Viridans and Enterococcus). Columbia Broth, in the presence of CO₂ and supplemented with the anticoagulant SPS (Sodium polyanetholsulfonate), is an excellent blood culture medium.
Peptone mixture and tryptic digest of beef heart provide nitrogen, vitamins, minerals and amino acids essential for growth. Dextrose is the fermentable carbohydrate providing carbon and energy. L-Cysteine HCl is the reducing agent. The medium is buffered with TRIS aminomethane and TRIS aminomethane HCl. Ferrous sulfate is added to facilitate organism growth. Magnesium sulfate is a magnesium ion required in a large variation of enzymatic reactions. Sodium chloride supplies essential electrolytes for transport and osmotic balance. Sodium carbonate is a neutralizer that will absorb any toxic metabolites.
Technical specifications
| Solubility | Without residues |
|---|---|
| Appearance | Fine powder |
| Color of the dehydrated medium | Beige |
| Color of the prepared medium | Clear amber, slightly opalescent |
| Final pH (25 °C) | 7.5 ± 0.2 |
| Amount per litre | 35 g |
| Storage | 2 to 25 °C |
Formula
| Ingredient | g/L |
|---|---|
| Dextrose | 2.5 |
| Ferrous sulfate | 0.02 |
| L-Cysteine hydrochloride | 0.1 |
| Peptone mixture | 20 |
| Sodium carbonate | 0.6 |
| Sodium chloride | 5 |
| Magnesium sulfate anhydrous | 0.1 |
| Tryptic digest of beef heart | 3 |
| TRIS hydroxymethyl aminomethane HCl | 2.86 |
| TRIS aminomethane | 0.83 |
Preparation and use
Preparation
Suspend 35 grams of the medium in one liter of distilled water. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Dispense into appropriate containers and sterilize in autoclave at 121 °C for 15 minutes.
Instructions for use
Inoculate and incubate at 35 ± 2 °C for 18-48 hours.
Quality control
Microbiological test
Incubation conditions: 35 ± 2 °C / 18-48 h.
| Microorganism | Specification |
|---|---|
| Neisseria meningitidisATCC 13090 | Good growth |
| Streptococcus pyogenesATCC 19615 | Good growth |
| Staphylococcus aureusATCC 25923 | Good growth |
| Streptococcus pneumoniaeATCC 6305 | Good growth |
Bibliography
- J.A. Morello & P.D. Ellner (1969). New medium for blood cultures. (Appl. Microb. 17: 68-70)
- H.D. Isenberg (ed) (1992). Clinical microbiology procedures handbook, vol. 1 (American Soc. for Microbiol., Washington, D.C.)
- P.R. Murray, E.J. Baron, M.A. Pfaller, et al. Manual of clinical microbiology, 6th ed. (American Soc. for Microbiol., Washington, D.C.).
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