Actinomyces Broth

Intended Use

Actinomyces Broth is used as a liquid medium or, with the addition of 7 or 20 g/L of agar, as a semisolid or solid medium, respectively, for the maintenance or cultivation of Actinomyces species.

Summary and Explanation

Actinomyces Broth is a basic medium modified from the Actinomyces Maintenance Medium of Pine and Watson.¹ It is recommended for use in the growth and maintenance of members of the genus Actinomyces.²

Principles of the Procedure

Actinomyces Broth contains meat infusion, peptone, yeast extract, soluble starch, L-cysteine and dextrose, which provide carbon, nitrogen, sulfur, vitamins and other growth factors required for the metabolism of Actinomyces spp. The salts provide essential minerals and electrolytes.

User Quality Control

Identity Specifications
Actinomyces Broth
Dehydrated Appearance: Fine, homogeneous, free of extraneous material.
Solution:                       5.7% solution, soluble in purified water.
                                    Solution is light to medium, yellow to tan,
                                    trace hazy to moderately hazy.
Prepared Appearance:     Light to medium, yellow to tan, trace hazy
to moderately hazy.
Reaction of 5.7%
Solution at 25°C:            pH 6.9 ± 0.2
Cultural Response
Actinomyces Broth
Prepare the medium per label directions. Inoculate and incubate anaerobically at 35 ± 2°C for 7 days.

Formula

Actinomyces Broth
Approximate Formula* Per Liter
Heart Muscle, Infusion from (solids).............................. 2.0 g
Pancreatic Digest of Casein......................................... 17.0 g
Yeast Extract............................................................. 10.0 g
Sodium Chloride.......................................................... 5.0 g
Dipotassium Phosphate.............................................. 13.0 g
Monopotassium Phosphate........................................... 2.0 g
Dextrose.................................................................... 5.0 g
Ammonium Sulfate...................................................... 1.0 g
L-Cysteine HCl............................................................ 1.0 g
Soluble Starch............................................................. 1.0 g
Magnesium Sulfate...................................................... 0.2 g
Calcium Chloride........................................................ 0.01 g
*Adjusted and/or supplemented as required to meet performance criteria.

Directions for Preparation from Dehydrated Product

1. Dissolve 57 g of the powder in 1 L of purified water. Add agar, 7 or 20 g/L, if a semisolid or solid medium is desired.
2. If agar is added, heat with frequent agitation just until solution occurs.
3. Dispense and autoclave at 121°C for 10 minutes.
4. Test samples of the finished product for performance using stable, typical control cultures.

Procedure

Inoculate Actinomyces cultures into tubes containing broth, semisolid or solid media. The semisolid medium should be stabinoculated and the slanted medium should be inoculated over its entire surface. Incubate cultures at 35 ± 2°C in an anaerobic atmosphere (BD GasPak™ EZ anaerobic system, or alternative system for the cultivation of anaerobic microorganisms).

Expected Results

After growth is obtained, tubes containing broth may be frozen for long-term storage. Cultures grown in the semisolid medium can be refrigerated after growth has been obtained. Agar slant cultures are for use in a relatively short period of time.

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