Rogosa SL Agar

Formula Procedure
BBL™ Rice Extract Agar 1. For use in the detection of chlamydospore formation.
Approximate Formula* Per Liter Inoculate the culture by cutting slits into the agar with an
White Rice, Extract from (solids) ................................. 5.0 g
Agar ......................................................................... 20.0 g
inoculating needle. Cover the inoculated slits with sterile
*Adjusted and/or supplemented as required to meet performance criteria. coverslips. Seal the plates to avoid moisture loss and
incubate at room temperature for 24-48 hours and up to
Directions for Preparation from 14 days before discarding as negative.
Dehydrated Product 2. For use in the promotion of chromogenesis in T. rubrum.
1. Suspend 25 g of the powder in 1 L purified water. To Streak-inoculate tubed medium slants. Tighten caps after
promote chlamydospore formation, suspend 13 g of the inoculation and then loosen slightly. After incubation for
powder in 1 L of purified water. 2-3 days, caps should be retightened to prevent further
2. Add 10 mL polysorbate 80. Mix until a uniform suspension evaporation of water. Incubate tubes at room temperature
is obtained. for up to 14 days.
3. Heat with frequent agitation and boil for 1 minute to
completely dissolve the powder. Expected Results
4. Dispense and autoclave at 121°C for 15 minutes.
5. Test samples of the finished product for performance using
After 24-48 hours, most strains of C. albicans and
C. stellatoidea will have formed typical chlamydospores.3 Invert
Q
stable, typical control cultures. the plate and examine microscopically (100× magnification) for R
chlamydospore formation along the line of inoculation.
User Quality Control Growth of T. rubrum is pink to red on medium containing
dextrose and, therefore, it is distinguishable from
Identity Specifications
T. mentagrophytes.
BBL™ Rice Extract Agar
Dehydrated Appearance: Fine, homogeneous, free of extrane-
ous material. Limitation of the Procedure
Solution: 1.3% solution, soluble in purified Polysorbate 80 enhances chlamydospore production by
water upon boiling. Solution is pale, C. albicans and C. stellatoidea; however, it also enhances chlamy-
yellow to tan, moderately hazy to hazy. dospore formation in other Candida species. Therefore, it is
Prepared Appearance: Pale, yellow to tan, moderately hazy necessary to use additional media for species identification.4
to hazy.
Reaction of 1.3%
Solution at 25°C: pH 6.6 ± 0.2
References
1. Taschdjian. 1953. Mycologia 45:474.
2. Taschdjian. 1957. Mycologia 49:332.
Cultural Response 3. Cooper and Silva-Hutner. 1985. In Lennette, Balows, Hausler and Shadomy (ed.), Manual of
clinical microbiology, 4th ed. American Society for Microbiology, Washington, D.C.
BBL™ Rice Extract Agar 4. MacFaddin. 1985. Media for isolation-cultivation-identification-maintenance of medical bacteria,
vol. 1. Williams & Wilkins, Baltimore, Md.
Prepare the medium per label directions and test for chlamydospore pro-
duction. Using fresh cultures, streak two parallel lines approximately 1.5
cm long and 1.0 cm apart. Make an S-shaped streak lightly back and Availability
forth across the two parallel streak lines. Place a coverslip over the streak BBL™ Rice Extract Agar
marks. Incubate at 20-25°C for 3-5 days and examine microscopically. Cat. No. 211567 Dehydrated – 100 g
ORGANISM ATCC™ RECOVERY CHLAMYDOSPORES Europe
Candida albicans 10231 Good + Cat. No. 254420 Prepared Plates – Pkg. of 20*
Candida albicans 60193 Good _ *Store at 2-8°C.

Rogosa SL Agar • Rogosa SL Broth

Intended Use
Rogosa SL Agar and Rogosa SL Broth are used for cultivating
oral, vaginal and fecal lactobacilli.
Summary and Explanation
Rogosa SL Agar and Broth, also known as RMW Agar/Broth,
are a modification of media described by Rogosa, Mitchell
and Wiseman.1,2 These media are used for isolation, enumeration
and identification of lactobacilli in oral bacteriology, feces,
vaginal specimens and foodstuffs.3,4 The low pH and high ac-
etate concentrations effectively suppress other bacterial flora
allowing lactobacilli to flourish.
Principles of the Procedure
Peptone provides carbon and nitrogen. Yeast extract is a source
of trace elements, vitamins and amino acids. Dextrose,
arabinose and saccharose are carbohydrate sources that
provide carbon. Sodium acetate and ammonium citrate
inhibit streptococci, molds and other oral microbial flora and
restrict swarming. Monopotassium phosphate provides

483
 Section III
QR Rogosa SL Agar, cont.

User Quality Control Directions for Preparation from

Identity Specifications
Difco™ Rogosa SL Agar
Dehydrated Appearance:
Solution:
Prepared Appearance:
Reaction of 7.5% Solution
(with glacial acetic acid
and reboiling) at 25°C:
Difco Rogosa SL Broth
™
Beige, homogeneous with soft clumps.
7.5% solution, soluble in purified
water upon boiling. Solution is light
amber, slightly opalescent and may
have a slight precipitate.
Light amber, slightly opalescent.
pH 5.4 ± 0.2
Dehydrated Product
1. Suspend the powder in 1 L of purified water:
Difco™ Rogosa SL Agar – 75 g;
Difco™ Rogosa SL Broth – 59.7 g.
Mix thoroughly.
2. Heat with frequent agitation and boil for 1 minute to
completely dissolve the powder.
3. Add 1.32 mL of glacial acetic acid and mix thoroughly.
4. Boil for 2-3 minutes. DO NOT AUTOCLAVE.
5. Test samples of the finished product for performance using
stable, typical control cultures.

Dehydrated Appearance: Beige, appears moist, with soft

clumps. Procedure
Solution: 6.0% solution, soluble in purified See appropriate references for specific procedures.
water upon boiling. Solution is light
amber, clear to slightly opalescent. Expected Results
Prepared Appearance: Light amber, clear to slightly opalescent. Refer to appropriate references and procedures for results.
Reaction of 6.0% Solution
(with glacial acetic acid
and reboiling) at 25°C: pH 5.4 ± 0.2
Limitation of the Procedure
The salt in the formulation makes the media unsuitable for
Cultural Response isolation of dairy lactobacilli; e.g., L. lactis, L. bulgaricus and
Difco™ Rogosa SL Agar or Rogosa SL Broth L. helveticus.4
Prepare the medium per label directions. Inoculate the agar medium
using the pour plate technique and incubate at 35 ± 2°C for 40-48 References
hours. Inoculate the broth medium and incubate at 35 ± 2°C for 18-48 1. Rogosa, Mitchell and Wiseman. 1951. J. Bacteriol. 62:132.
hours. 2. Rogosa, Mitchell and Wiseman. 1951. J. Dental Res. 30:682.
3. Hall, Ledenbach and Flowers. 2001. In Downes and Ito (ed.), Compendium of methods for the
microbiological examination of foods, 4th ed. American Public Health Association, Washington,
ORGANISM ATCC™ INOCULUM CFU RECOVERY
D.C.
Lactobacillus rhamnosus 9595 102-103 Good 4. MacFaddin. 1985. Media for isolation-cultivation-identification-maintenance of medical bacteria,
vol. 1. Williams & Wilkins, Baltimore, Md.
Lactobacillus delbrueckii
subsp. lactis 4797 102-103 Good
Staphylococcus aureus 25923 103-2×103 Marked to
Availability
complete inhibition Difco™ Rogosa SL Agar
Cat. No. 248020 Dehydrated – 500 g*

buffering capability. Magnesium sulfate, manganese sulfate and Japan

Cat. No. 251542 Prepared Plates – Ctn. of 100*
ferrous sulfate are sources of inorganic ions. Polysorbate 80
Difco Rogosa SL Broth
™
acts as a surfactant. Agar is the solidifying agent.
COMPF
Cat. No. 247810 Dehydrated – 500 g*
Formulae *Store at 2-8°C.
Difco™ Rogosa SL Agar
Approximate Formula* Per Liter
Tryptone ................................................................... 10.0 g
Yeast Extract .............................................................. 5.0 g
Dextrose ................................................................... 10.0 g
Arabinose ................................................................... 5.0 g
Saccharose ................................................................. 5.0 g
Sodium Acetate ....................................................... 15.0 g
Ammonium Citrate .................................................... 2.0 g
Monopotassium Phosphate ........................................ 6.0 g
Magnesium Sulfate .................................................... 0.57 g
Manganese Sulfate .................................................... 0.12 g
Ferrous Sulfate ........................................................... 0.03 g
Polysorbate 80 ........................................................... 1.0 g
Agar ......................................................................... 15.0 g
Difco™ Rogosa SL Broth
Consists of the same ingredients without the agar.
*Adjusted and/or supplemented as required to meet performance criteria.

484