Health Canada
www.hc-sc.gc.ca
Common menu bar links
Institutional links
Appendix G
Media Glossary
January 2006
Glossary of agars, broths and other reagents
Section C
Help on accessing alternative formats, such as Portable Document Format (PDF), MP3 and WAV files, can be obtained in the alternate format help section.
(PDF Version - 22 K)| Basal Medium | |
|---|---|
| Meat extract | 10.0 g |
| Peptone | 10.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Bacteriological charcoal | 4.0 g |
| Casein hydrolysate | 3.0 g |
| Sodium deoxycholate | 1.0 g |
| Ferrous sulphate (FeSO4) | 0.25 g |
| Sodium pyruvate | 0.25 g |
| Agar | 12.0 g |
| Distilled water | 1.0 L |
| pH 7.5 ± 0.2 | |
| Supplements | |
| Sodium cefoperazone | 0.032 g |
| Amphotericin B | 0.010 |
| Distilled water | 10.0 mL |
Complete Medium:
Dissolve all components, except supplement, in 1.0 L of distilled water and boil to dissolve completely. Autoclave at 121oC for 15 min. Cool to 50oC. Add components for the supplement to distilled water. Mix thoroughly and filter sterilize using a 0.2 µm filter and add aseptically to the medium. Mix well and pour into sterile Petri plates.
| Basal Medium | |
|---|---|
| Peptone | 10.0 g |
| Beef Extract | 1.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Bromocresol purple | 0.02 g |
| Agar | 15.0 g |
| Distilled water | 950 mL |
| pH 6.8 ± 0.2 | |
| Supplements (Rhamnose, Mannitol and Xylose) | |
| Carbohydrate | 20.0 g |
| Distilled Water | 100.0 mL |
Complete Medium:
The basal medium is commercially available as Purple Agar Base. Add 31.0 g of purple agar base to distilled water and mix thoroughly. Sterilize at 121oC for 15 minutes. Filter sterilize 100 mL each of 20% aqueous solutions of rhamnose, mannitol and xylose. Temper basal medium, add 50.0 mL of the sterile carbohydrate solution, then pour thick plates. Plates can be stored for at least 2-3 weeks at 4oC. Longer storage times must be validated by the individual lab.
| Basal Medium | |
|---|---|
| Peptone | 10.0 g |
| Beef extract | 1.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Bromocresol purple | 0.02 g |
| Distilled Water | 950 mL |
| pH 6.8 ± 0.2 | |
| Supplements (Rhamnose, Mannitol and Xylose) | |
| Carbohydrate | 5.0 g |
| Distilled water | 100 mL |
Complete Medium:
The basal medium is commercially available as Purple Broth Base. Add 16.0 g of purple broth base to distilled water and mix thoroughly. Dispense 9.0 mL portions in 16 x 125 mm tubes each containing a Durham tube. Autoclave at 121oC for 15 min. Prepare all carbohydrates, except esculin, as 5% solutions and filter sterilize. Add 1.0 mL carbohydrate solution to 9.0 mL broth base to yield a final concentration of 0.5% carbohydrate in broth.
Note: Add esculin directly into base broth to make a 0.5% solution and autoclave at 115oC for 15 min. A 5% solution of esculin at room temperature is a gel that cannot be pipetted.
| Basal Medium | |
|---|---|
| Sodium Chloride (NaCl) | 5.0 g |
| Sodium thioglycolate | 1.5 g |
| Sodium phosphate, dibasic (Na2HPO4) | 1.1 g |
| Agar | 5.0 g |
| Distilled water | 1.0 L |
| pH 8.4 ± 0.2 | |
| Supplements | |
| Calcium chloride (CaCl2) | 0.1 g |
| Distilled water | 10.0 mL |
Complete Medium:
Suspend ingredients, except CaCl 2, in water. Heat to boiling to dissolve completely and cool to 50oC. Add 9.0 mL of filter sterilized 1% CaCl 2 . Dispense in 6.0-8.0 mL capacity screw-cap tubes to within 5.0 mm of the top. Autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Casamino acids | 30.0 g |
| Yeast extract | 4.0 g |
| Potassium phosphate, dibasic ( K2HPO4) | 0.5 g |
| Distilled water | 1.0 L |
| pH 7.4 ± 0.2 | |
Complete Medium:
Dissolve ingredients in water. Adjust pH and dispense 10.0 mL into 50.0 mL flasks. Autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Pancreatic digest of gelatin | 20.0 g |
| Potassium sulfate (K2SO4) | 10.0 g |
| Magnesium chloride (MgCl 2) | 1.4 g |
| Cetrimide | 0.3 g |
| Glycerol | 10.0 mL |
| Agar | 13.6 g |
| Distilled water | 1.0 L |
| pH 7.2 ± 0.2 | |
Complete Medium:
Add components to distilled water. Gently heat and bring to a boil. Autoclave for 15 min at 118oC and 13 psi pressure. Dispense.
Coagulase Plasma
Certified rabbit plasma containing EDTA is commercially available. Reconstitute as directed by the manufacturers. The reconstituted plasma may be kept in the refrigerator for up to 14 days without loss of potency. It is not satisfactory for use if gross contamination occurs. After being kept in the refrigerator, the plasma solution is cold enough to delay clotting for 10-15 min. This delay can be prevented by warming the plasma solution to 35oC before use.
If dehydrated product is not available, use fresh rabbit plasma collected aseptically in containers with EDTA (1.0 mL of a 15% solution of the potassium salt per 100 mL blood). Dilute plasma 1:2 or 1:3 with sterile distilled water and test each batch with coagulase-positive and coagulase-negative strains of staphylococci before putting it into routine use.
| Basal Medium | |
|---|---|
| Beef heart | 454.0 g |
| Proteose peptone | 20.0 g |
| Dextrose | 2.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| pH 7.2 ± 0.2 | |
Complete Medium:
The medium may be prepared by distributing 2.5 g of a prepared and dehydrated medium into 20 X 150 mm screw-cap culture tubes; add 20.0 mL of cold distilled water; mix thoroughly, and let stand to ensure thorough wetting of all particles. Sterilize for 15 min at 121oC or the equivalent. Normally the prepared sterilized media can be maintained unaffected under refrigeration (0oC to 4oC) for up to 4 weeks. Prior to use, the refrigerated medium should be deaerated at 100oC for 10 min and cooled before use.
| Basal Medium | |
|---|---|
| Solution A | |
| Crystal violet (85-90% dye content) | 2.0 g |
| Ethyl alcohol (95%) | 20.0 mL |
| Solution B | |
| Ammonium oxalate | 0.8 g |
| Distilled water | 80.0 mL |
Complete Medium:
Dissolve the crystal violet in the alcohol and the ammonium oxalate in the distilled water. Mix the two solutions and store the mixture for 24 hours before use.
