IX. IDENTIFICATION OF NIYCOPLASMAS
3. Mediuni S U : Shepard meditun (for Ureaplnsnia spp.)
(a) Liquid medium
Trypticase soy broth powder (Baltimore Biological Laboratories)
(3% (w/v) solution)
Horse serum (unheated)
Yeast extract (25%) (Taylor-Robinson et al., 1963)
Urea (40% (w/v) solution)
Phenol red (0.06% (w/v) solution)
Penicillin (20 000 I U ml-1)
Adjust p H to 6.0.
(b) Solid medium
This is prepared by adding 1.4 g Purified Agar (Code L 28 Oxoid).
4. Medium BAC Y (for M . faucium and M . lipophilum)
(a) Liquid medium
Heart infusion broth (Difco)
(Sterilise by autoclaving)
Horse serum (unheated)
Yeast extract (25%) (Taylor-Robinson et al., 1963)
Deoxyribonucleic acid (Sigma, 0.2% (w/v) solution)
Cystein hydrochloride (10% (w/v) solution)
L-arginine (30% (w/v) solution)
Phenol red (0.06% (w/v) solution)
Thallium acetate (1 .O% (w/v) solution)
Penicillin (20 000 I U ml-1)
Adjust pH to 7.3
(b) Solid medium
Replace heart infusion broth with heart infusion agar (Difco).
5 . Medium F: Frey medium (for M . synoviae)
(a) Liquid medium
Bacto PPLO broth w/o CV (Difco)
Distilled water
Eagle’s essential vitamins ( x loo)?
Glucose (50% (w/v) solution)
Swine serum (inactivated 56°C 30 min-I)
B-NADHf (1 % (w/v) solution)
Cystein hydrochloride (1 % (w/v) solution)
Phenol red (0.06% (w/v) solution)
Penicillin (20 000 I U ml-1)
The solutions of @-NADH and cystein hydrochloride are mixed,
and after 10 min added to the other ingredients.
Adjust p H to 7.8.
43 1
100.0 ml
22.0 ml
10.0 ml
1.6 ml
5.0 ml
1.0 ml
90.0 ml
20.0 ml
10.0 ml
1.3 ml
1 a24 ml
4.25 ml
5.0 ml
1.0 ml
0.25 ml
2.25 g
90.0 ml
0.025 ml
2.0 ml
12.0 ml
1.0 ml
1 .O ml
5.0 ml
0.25 ml
t Therkelsen (1961).
3 8-Nicotinamide dinucleotide, reduced form, Sigma, Grade 111.