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Appendix G
Media Glossary
January 2006
Glossary of agars, broths and other reagents
Section B
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(PDF Version - 26 K)| Basal Medium | |
|---|---|
| Tryptone | 10.0 g |
| Beef extract | 5.0 g |
| Yeast extract | 1.0 g |
| Glycine | 12.0 g |
| Lithium chloride | 5.0 g |
| Sodium pyruvate | 10.0 g |
| Agar | 20.0 g |
| Distilled water | 1.0 L |
| pH 7.0 ± 0.2 | |
| Supplements | |
| Filter-sterilized 1% potassium tellurite solution | |
| Egg yolk emulsion (50%) | |
Complete Medium:
Add components to water. Gently heat and bring to a boil. Autoclave at 121oC for 15 min. Cool to 45-50oC and dispense.
Egg yolk tellurite emulsion is commercially available and usually contains about 50% egg yolk. Use as per manufacturer's instructions.*
* Example: Aseptically add 50.0 mL of pre-warmed (45-50oC) commercial egg yolk/tellurite emulsion to the basal medium. Mix thoroughly and dispense.
Note: The surface of the agar should be dried before inoculation. It has been observed that freshly prepared BP medium may be toxic to injured cells. It is therefore advisable to store the plates at refrigeration temperature overnight before inoculation. The medium should be opaque; do not use non-opaque plates. BP media can have a shelf-life of one to several weeks under specified storage conditions. However, the selectivity and diagnostic characteristics of the components in the complete medium may deteriorate with time, especially if the agar plates are not prepared from good quality, commercially-available products. It is the responsibility of each laboratory to validate the storage time appropriate for their BP agar plates, based on the ability of the product to meet specifications for selectivity, recoverability, and expected performance.
| Basal Medium | |
|---|---|
| Peptone | 10.0 g |
| Beef extract | 5.0 g |
| Dextrose | 5.0 g |
| Disodium phosphate (Na 2HPO 4) | 4.0 g |
| Ferrous sulfate (FeSO 4) | 0.3 g |
| Bismuth sulfite indicator | 8.0 g |
| Brilliant green | 0.025 g |
| Agar | 20.0 g |
| Distilled water | 1.0 L |
| pH 7.6 ± 0.2 | |
Complete Medium:
Suspend ingredients in one liter of distilled water and mix thoroughly. Uniformly heat the medium to incipient boiling using a hotplate rather than an open flame. Do not autoclave and do not overheat. Frequently swirl medium during heating. Cool to 45-50oC and swirl medium to evenly disperse the heavy precipitate just prior to pouring the medium into plates.
Note: Before use, store prepared plates overnight in a darkened refrigerator. Plates can be stored for up to 4 days. If uninoculated plates begin to turn a darker green, they are to be discarded. This change in color is indicative of oxidation. The probability and degree of oxidation increases with increased storage time and is accelerated by light exposure. Such plates may be inhibitory to some salmonellae.
| Basal Medium | |
|---|---|
| Trypticase | 10.0 g |
| Beef extract | 3.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Agar | 15.0 g |
| Distilled water | 1.0 L |
| pH 7.2 ± 0.1 | |
| Supplement | |
| Sterile defibrinated blood | 50.0 mL |
Complete Medium:
Suspend ingredients in 1.0 L distilled water. Heat to boiling to dissolve ingredients. Sterilize at 121oC for 15 min. Cool to 45-50oC and aseptically add sterile defibrinated blood. Mix thoroughly but avoid incorporation of air bubbles. Dispense. The solidified, complete medium cannot be re-liquified.
Blood Agar- Listeria
Prepare blood agar plates as soon as possible after receiving fresh blood, using blood agar base or preferably Trypticase Soy agar with 7% defibrinated horse blood. Rehydrate and sterilize as recommended by the manufacturer. Agar and blood should both be at 45-50oC before combining and pouring plates. Plates can be stored at 4oC for 1 month. Sheep blood plates for the CAMP test are prepared and stored in a similar way
| Basal Medium | |
|---|---|
| Calf brain, infusion from | 200.0 g |
| Beef heart, infusion from | 250.0 g |
| Proteose peptone | 10.0 g |
| Dextrose | 2.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Sodium phosphate (Na 2HPO 4) | 2.5 g |
| Distilled water | 1.0 L |
| pH 7.4 ± 0.2 | |
Complete Medium:
Dissolve ingredients in 1.0 L of distilled water. Distribute as required (tubes or flasks) and sterilize at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Calf brain, infusion from | 200.0 g |
| Beef heart, infusion from | 250.0 g |
| Proteose peptone | 10.0 g |
| Dextrose | 2.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Sodium phosphate (Na 2HPO 4) | 2.5 g |
| Yeast extract | 6.0 g |
| Distilled water | 1.0 L |
| pH 7.4 ± 0.2 | |
Complete Medium:
Dissolve ingredients in 1.0 L of distilled water. Distribute as required (tubes or flasks) and sterilize at 121oC for 15 min. Add yeast extract to BHI before autoclaving.
| Basal Medium | |
|---|---|
| Peptone | 10.0 g |
| Lactose | 10.0 g |
| Ox-gall | 20.0 g |
| Brilliant green | 0.0133 g |
| pH 7.2 ± 0.2 | |
Complete Medium:
Dissolve the peptone and lactose in 500 mL of water and add the ox-gall dissolved in 200 mL of water. Bring the volume to approximately 975 mL with water and adjust pH. Add 13.3 mL of a 1% aqueous solution of brilliant green, bring the total volume to 1.0 L, stir and if necessary filter through cotton to clarify. Dispense in 10.0 mL volumes into tubes containing gas vials and sterilize at 121oC for 12 min.
| Basal Medium | |
|---|---|
| Yeast extract | 3.0 g |
| Peptone | 10.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Lactose | 10.0 g |
| Sucrose | 10.0 g |
| Phenol red | 0.08 g |
| Brilliant green | 0.0125 g |
| Sodium sulfapyridine | 1.0 g |
| Agar | 20.0 g |
| Distilled water | 1.0 L |
| pH 6.9 ± 0.2 | |
Complete Medium:
Suspend the ingredients in 1 L of distilled water. Heat to boiling with frequent swirling. Autoclave at 121oC for 15 min and cool to 45-50oC. Dispense medium into plates and allow to dry with lids partially removed.
| Basal Medium | |
|---|---|
| Brilliant green dye | 1.0 g |
| Sterile distilled water | 100 mL |
Complete Medium:
Dissolve 1.0 g brilliant green dye in 100 mL sterile water (1% w/v). Add 2.0 mL of the 1% brilliant green solution to 1.0 L of sterile water (0.002% w/v).
| Basal Medium | |
|---|---|
| Tryptone | 10.0 g |
| Peptic digest of animal tissues | 10.0 g |
| Dextrose | 1.0 g |
| Yeast extract | 2.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Sodium bisulfite (NaHSO 3) | 0.1 g |
| Distilled water | 1.0 L |
| pH 7.0 ± 0.2 | |
Complete Medium:
Suspend ingredients in 1.0 L of distilled water and boil to dissolve completely. Autoclave at 121oC for 15 min and dispense.
| Basal Medium | |
|---|---|
| Tryptone | 10.0 g |
| Peptic digest of animal tissues | 10.0 g |
| Dextrose | 1.0 g |
| Yeast extract | 2.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Sodium bisulfite (NaHSO 3) | 0.1 g |
| Neutral Red | 0.02 g |
| Agar | 1.6 g |
| Distilled water | 1.0 L |
| pH 7.0 ± 0.2 | |
Complete Medium:
Mix the above ingredients in distilled water and heat with frequent agitation. Boil for about 1 min. Dispense 5.0-7.0 mL into screw-cap tubes (16 x 125 mm) and autoclave at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Peptone | 5.0 g |
| Glucose | 5.0 g |
| Dipotassium phosphate (K 2HPO 4) | 5.0 g |
| Distilled water | 1.0 L |
| pH 7.5 ± 0.2 | |
Complete Medium:
Add ingredients to 1.0 L of distilled water, heat gently with stirring to obtain complete solution. Dispense into test tubes and sterilize at 121oC for 15 min.
| Basal Medium | |
|---|---|
| Peptone | 5.0 g |
| Glucose | 5.0 g |
| Dipotassium phosphate (K 2HPO 4) | 5.0 g |
| Sodium chloride (NaCl) | 2.0 - 3.0 g |
| Distilled water | 1.0 L |
| pH 7.5 ± 0.2 | |
Complete Medium:
Add ingredients to 1.0 L of distilled water, heat gently with stirring to obtain complete solution. Dispense into test tubes and sterilize at 121oC for 15 min. (For halophilic Vibrio species)
| Basal Medium | |
|---|---|
| Peptone | 10.0 g |
| Sodium chloride (NaCl) | 5.0 g |
| Disodium phosphate | 3.5 g |
| Potassium dihydrogen phosphate | 1.5 g |
| Distilled water | 1.0 L |
| Final pH 7.2 ± 0.2 | |
Complete Medium:
Dissolve the ingredients in 1.0 L of distilled water. Dispense into appropriate glassware and autoclave at 121oC for 15 min.
