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Material Safety Data Sheet

Optional Products

Organisms

Organisms in the Clinical sector this product works with:

Dehydrated Culture Media

Sector: Clinical

MYCOPLASMA BROTH BASE

Code: CM0403

A basic medium which, after enrichment with a supplement can be used in the isolation and cultivation of mycoplasmas from clinical specimens.

Typical Formula*

gm/litre

Bacteriological peptone

10.0

Lab-Lemco’ powder

10.0

Sodium chloride

5.0

Mineral supplement

0.5

pH 7.8 ± 0.2 @ 25°C

 

* Adjusted as required to meet performance standards 

Directions
Dissolve 25.5g in 1 litre of distilled water. Mix well and distribute in 80ml volumes. Sterilise by autoclaving at 121°C for 15 minutes. Cool to approximately 50°C and add one vial of Mycoplasma Supplement SR0059 reconstituted as directed.

MYCOPLASMA SUPPLEMENT - G SELECTIVE SUPPLEMENT

Code: SR0059

POISON - CONTAINS THALLIUM SALT

Vial contents (each vial is sufficient for 80 ml of medium)

per vial
per litre

Horse serum

20.0ml

200.0ml

Yeast extract (25% w/v)

10.0ml

100.0ml

Thallous acetate

25.0mg

250.0mg

Penicillin

20,000IU

200,000IU

Description
Oxoid Mycoplasma Broth Base complements Oxoid Mycoplasma Agar Base CM0401.

It requires supplementation with yeast extract, serum and antibiotics, which are available as Mycoplasma Supplement-G SR0059.

Carbohydrate fermentation by mycoplasmas can be tested by incorporating 1% w/v carbohydrate and 0.005% w/v phenol red into the broth medium.

A Ureaplasma broth1 can be prepared by adding to 95 ml broth medium (pH adjusted to 6.0):

Vial contents

Horse Serum

4.0ml

Urea

0.05g

Phenol red

0.001g

Penicillin

100,000 units

A similar medium was described by Taylor-Robinson et al.2 where reference is made to the use of HEPES buffer to induce large colony-forming Ureaplasma strains and for the isolation and titration of viable mycoplasma by the metabolism of arginine or glucose and measuring the pH change in the medium.

A selective medium for Mycoplasma pneumoniae was described by Kraybill & Crawford3.

Most strains of mycoplasmas are encouraged by growth in a biphasic medium in which a layer of Mycoplasma Broth Base covers a basal layer of Mycoplasma Agar Base CM0401. Both broth and agar layers should be supplemented with Mycoplasma Supplement-G SR0059. Inclusion of the supplement provides the necessary factors for growth and prevents overgrowth of slow growing contaminating organisms.    

Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the prepared medium at 2-8°C.

Appearance
Dehydrated medium: Straw coloured, free-flowing powder
Prepared medium: Straw coloured solution

Quality control

Positive control:

Expected result at 35°C

Mycoplasma pneumoniae ATCC® 15531

Slightly turbid

Negative control:

 

Escherichia coli ATCC® 25922 *

Inhibited
* This organism is available as a Culti-Loop®

Precautions
Thallous acetate is toxic, observe the precautions stated under Hazards section..
Sub-culture the broth to agar as soon as any indicator added begins to change colour before the pH change destroys the organism.

References
1. Shepard M. C. and Lanceford C. D. (1970) Appl. Microbiol. 20. 539-543.
2. Taylor-Robinson D., Martin-Bourgon C., Watanable T. and Addey J. P. (1971) J. Gen. Microbiol. 68. 97-107.
3. Kraybill W. H. and Crawford Y. E. (1965) Proc. Soc. Exp. Biol. Med. 118. 965-967.

 
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