IX. IDENTIFICATION OF MYCOPLASMAS
433
The suspension is heated to 37°C for 20 min, thereafter heated to
90-100°C for 5 min. After cooling, centrifuge at 100 x g for
,30 min. The supernatant is dispensed in appropriate volumcs
and autoclaved at 115 "C for 2-5 min. Stored at - 20°C.
B. Media for biochemical tests
1. Glucose fermentation
(a) Basal medium: medium bi
Heart infusion broth (Difco)
Sterilise by autoclaving
PPLO serum fraction (Difco)
Deoxyribonucleic acid (Sigma, 0.20/6 (w/v) solution)
Phenol red (0.06% (w/v) solution)
Thallium acetate (1 % (w/v) solution)
Penicillin (20 000 I U ml-1)
Adjust pH to 7.8
(b) Test medium: medium bg
Medium b i
Glucose (50% (w/v) solution)
Adjust p H to 7.8
enzyme-treated see page 398
2. Arginine hydrolysis
(a) Basal medium: medium bz
As bl, except that pH is adjusted to 7.3
(b) Test medium: medium ba
Medium bz
L-arginine (30% (w/v) solution)
Adjust pH to 7.3
3. Urea hydrolysis
Test medium: medium SU (as A.3) with pH adjusted to 6.8
4. Phosphatase activity
Test medium: medium Bph
Heart infusion agar (Difco)
(Sterilise by autoclaving)
Horse serum (heated at 60°C for 60 min)
Yeast extract (25%) (Taylor-Robinson et d., 1963)
Sodium phenolphthalein diphosphate (1 yo (w/v) solution)
Penicillin (20 000 I U ml-l)
Thallium acetate (1 76 (w/v) solution)
Adjust pH to 7.8
15
120.0 ml
1.0 ml
1.2 ml
5.0 ml
1 .O ml
0-25 ml
128.0 ml
1.6 ml
128.0 ml
4.25 ml
74.0 ml
20.0 ml
5.0 ml
1 .O ml
0.2 ml
1 .O ml