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Appendix G
Media Glossary
January 2006
Glossary of agars, broths and other reagents
Section H
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(PDF Version - 29 K)| Basal Medium | |
|---|---|
| Beef heart infusion from 500 g | 1.0 L |
| Tryptose | 10.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| pH 7.4 ± 0.2 | |
Complete Medium:
Broth: Suspend basal medium ingredients in 1.0 L of distilled water and dispense into tubes. Autoclave at 121oC for 15 min.
*For halophilic Vibrio species add an extra 15.0 g/L NaCl.
Agar: Suspend basal medium ingredients and 15.0 g of agar in 1.0 L of distilled water. Heat to boiling with constant stirring to dissolve all ingredients. Autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Lactose | 12.0 g |
| Peptic digest of animal tissue | 12.0 g |
| Sucrose | 12.0 g |
| Bile salts | 9.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Sodium thiosulphate (Na2S2O3) | 5.0 g |
| Yeast extract | 3.0 g |
| Salicin | 2.0 g |
| Ferric ammonium citrate | 1.5 g |
| Acid fuchsin | 0.1 g |
| Bromthymol blue | 0.064 g |
| Agar | 13.5 g |
| Distilled water | 1.0 L |
| pH 7.6 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling with constant stirring to dissolve all ingredients. Do not autoclave. Dispense into sterile Petri dishes.
Caution: Acid fuchsin is a potential carcinogen and care must be taken to avoid inhalation of the powdered dye and contact with skin. Weat suitable protective clothing.
| Basal Medium | |
|---|---|
| Columbia blood agar base | 39.0 g |
| Lithium Chloride (LiCl) | 7.5 g |
| Ferric ammonium citrate | 0.5 g |
| Colistin methane sulfonate (11.500 I.U./mg) | 10.0 mg |
| Agar | 5.0 g |
| Distilled water | 1.0 L |
| Horse blood | |
| pH 7.0 ± 0.2 | |
| Antibiotic supplement | |
| Ceftazidime acid pentahydrate | 400 mg |
| 0.1 M phosphate buffer (pH 7.0) | 100 mL |
Complete Medium
Note : This medium is a modification of MOX, with a non-selective horse blood overlay agar.
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling with constant stirring to dissolve all ingredients. Autoclave at 121oC for 15 min and cool to 45-50oC. Add 5.0 mL of the antibiotic solution to the cooled basal medium to give a final concentration of 20.0 mg/L. Plates are made with a 10.0 mL base layer (without horse blood), and soon after solidifying are overlayed with 5.0 mL of the same agar to which 4% horse blood (tempered to 20oC) has been added.
Antibiotic Solution: Dissolve the ceftazidime in the phosphate buffer and sterilize by filtration. Store in 5.0 mL portions at 4oC.
*Poysky, F.T., R.N. Paranjpye, L.C. Lashbrook., M.E. Peterson, G.A. Pelroy and M.W. Eklund. 1993. Selective and differential medium for isolation of Listeria monocytogenes from foods. J. Food Prot. 56:326-329, 332.
| Basal Medium | |
|---|---|
| Tryptone | 20.0 g |
| Bile salts no. 3 | 1.12 g |
| Sodium chloride (NaCl) | 5.0 g |
| Sorbitol | 20.0 g |
| 4-methylumbelliferyl-ß-D-glucoronide (MUG) | 0.1 g |
| Bromocresol purple | 0.015 g |
| Agar | 15.0 g |
| Distilled water | 1.0 L |
| pH 7.2 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling with constant stirring to dissolve all ingredients. Autoclave at 121oC for 15 min.
Note : A different medium designated as HC Agar Base is used for the isolation of molds from cosmetic products.
Note : Media can be made with or without the addition of MUG.
| Sodium hippurate solution | |
|---|---|
| Sodium hippurate | 1.25 g |
| Distilled water | 50.0 mL |
| Ninhydrin solution (prepare weekly) | |
| Ninhydrin | 1.5 g |
| Acetone | 25.0 mL |
| Ethanol (95%) | 25.0 mL |
Complete Medium:
Sodium hippurate solution: Dissolve and filter sterile through a membrane (pore size 0.2µm). Dispense in 0.5 mL aliquots in small tubes. Cork or screw cap and store et -20oC.
Ninhydrin solution: Store in a brown bottle at 4oC.
| Basal Medium | |
|---|---|
| Peptone | 2.0 g |
| Yeast extract | 0.5 g |
| Sodium chloride (NaCl) | 30.0 g |
| Glucose | 10.0 g |
| Bromocresol purple | 0.015 g |
| Agar | 3.0 g |
| Distilled water | 1.0 L |
| pH 7.4 ± 0.2 | |
Complete Medium:
Suspend basal medium ingredients in 1.0 L of distilled water. Heat to boiling with constant stirring to dissolve all ingredients. Dispense and autoclave at 121oC for 15 min .
