OGYE Agar Base

Directions for Preparation from Dehydrated Product

1. Suspend 9.4 g of the powder in 1 L of purified water. Mix thoroughly. 2. Heat with frequent agitation and boil for 1 minute to completely dissolve the powder. 3. Autoclave at 121C for 15 minutes. Add 1% carbohydrate before or after autoclaving depending on heat lability. 4. Test samples of the finished product for performance using stable, typical control cultures.

Limitations of the Procedure

1. The acid reaction produced by oxidative organisms is apparent at the surface and gradually spreads throughout the medium. If the oxidation is weak or slow, however, an initial alkaline reaction at the surface of the open tube may perist for several days and eventually convert to an acid reaction. 2. If an organism is unable to grow on OF Basal Medium, Cowan5 recommends adding either 2% serum or 0.1% yeast extract to each carbohydrate tube.

Procedure
Inoculate a pair of OF tubes of each carbohydrate used with each organism being tested. The tubes should be stabbed to approximately 1/4 inch from the bottom using an inoculating needle and a light inoculum. Overlay one tube of each pair with sterile mineral oil. Incubate tubes at 35 2C in an aerobic atmosphere for 48 hours. Do not discard as negative until after 4 days of incubation.

References
1. Hugh and Leifson. 1953. J. Bacteriol. 66:24. 2. MacFaddin. 2000. Biochemical tests for identification of medical bacteria, 3rd ed. Lippincott Williams & Wilkins, Baltimore, Md. 3. MacFaddin. 1985. Media for isolation-cultivation-identification-maintenance of medical bacteria, vol. 1. Williams & Wilkins, Baltimore, Md. 4. Shigei. 1992. In Isenberg (ed.), Clinical microbiology procedures handbook, vol. 1. American Society for Microbiology, Washington, D.C. 5. Cowan. 1974. Cowan and Steeles manual for the identification of medical bacteria, 2nd ed. Cambridge University Press, Cambridge, Mass.

O P

Availability
Difco OF Basal Medium
BAM USDA

Expected Results
Record results as acid (A) or alkaline/no change (). Also record whether or not the organism is motile as evidenced by the appearance of growth away from the line of inoculation. Typical reaction patterns are as follows.2-4
Enteric OF Carbohydrate Utilization Patterns
REACTION TUBE WITH REACTION OPEN TUBE COVERED TUBE

Cat. No. Cat. No. Cat. No.

268820 221326 297783 221328 221329 297784 297366 297785 221330 221332 221334 297786 296374 297368 297365 297367 221336 221338 266631

Dehydrated 500 g Prepared Tubes, 5 mL (K Tubes) Pkg. of 10* Prepared Tubes with Arabinose Pkg. of 10* Prepared Tubes with Dextrose Pkg. of 10* Prepared Tubes with Dextrose Ctn. of 100* Prepared Tubes with Dulcitol Pkg. of 10* Prepared Tubes with Fructose (Levulose) Pkg. of 10* Prepared Tubes with Galactose Pkg. of 10* Prepared Tubes with Lactose Pkg. of 10* Prepared Tubes with Maltose Pkg. of 10* Prepared Tubes with Mannitol Pkg. of 10* Prepared Tubes with Mannose Pkg. of 10* Prepared Tubes with Raffinose Pkg. of 10* Prepared Tubes with Rhamnose Pkg. of 10* Prepared Tubes with Salicin Pkg. of 10* Prepared Tubes with Sorbitol Pkg. of 10* Prepared Tubes with Sucrose Pkg. of 10* Prepared Tubes with Xylose Pkg. of 10* Bottle 60 mL

BBL OF Basal Medium BBL OF Basal Medium with Carbohydrates

Oxidation (O) Fermentation (F) Anaerogenic Aerogenic Neither Oxidation nor Fermentation () Both Oxidation and Fermentation (O/F)

Open Covered Covered

Yellow (A) Yellow (A) Yellow (AG)

Green () Yellow (A) Yellow (AG) Green ()

Neither* Blue or Green () Both

Yellow (A or AG) Yellow (A or AG)

A = acid production G = gas production = no change or alkaline * = Uninoculated carbohydrate control reading; no change in color.

Pasco Mineral Oil

Cat. No.
*Store at 2-8C.

OFPBL Agar
(See PC Agar)

OGYE Agar Base Antimicrobic Vial Oxytetracycline

Intended Use
OGYE Agar Base is for use with Antimicrobic Vial Oxytetracycline in isolating and enumerating yeasts and molds in foods.

Summary and Explanation

Acidified agar may be used for enumerating yeasts and molds in foods and dairy products. However, in some cases, antimicrobics better suppress bacterial growth and improve recovery of yeasts and molds.1,2 411

Section III OP OGYE Agar Base, cont.

Mossel et al.3,4 described Oxytetracycline-Glucose Yeast Extract (OGYE or OGY) Agar for selectively isolating and enumerating yeasts and molds in foods. Mossel et al. demonstrated improved recovery compared to acidified agar media. OGYE Agar is specified as a standard methods medium for use with dairy products.1

User Quality Control

Identity Specifications
Difco OGYE Agar Base
Dehydrated Appearance: Solution: Tan, free-flowing, homogeneous. 3.7% solution, soluble in purified water upon boiling. Solution is medium amber, very slightly opalescent. Medium amber, slightly opalescent. pH 7.0 0.2 Yellow cake or powder

Principles of the Procedure

OGYE Agar Base contains yeast extract to supply B-complex vitamins which stimulate bacterial growth. Dextrose is the carbon energy source. Agar is the solidifying agent. Antimicrobic Vial Oxytetracycline inhibits the growth of bacteria.
Prepared Appearance: Reaction of 3.7% Solution at 25C:

Difco Antimicrobic Vial Oxytetracycline

Lyophilized Appearance:

Formulae
Difco OGYE Agar Base
Approximate Formula* Per Liter Yeast Extract .............................................................. 5.0 Dextrose ................................................................... 20.0 Agar ......................................................................... 12.0 g g g

Cultural Response
Difco OGYE Agar Base
Prepare the medium per label directions (with the addition of Antimicrobic Vial Oxytetracycline). Inoculate using the pour plate technique and incubate at 22 3C for 48-72 hours (up to 5 days if necessary).
ORGANISM ATCC INOCULUM CFU RECOVERY

Difco Antimicrobic Vial Oxytetracycline

Formula Per 10 mL Vial Oxytetracycline ....................................................... 100
*Adjusted and/or supplemented as required to meet performance criteria.

mg

Directions for Preparation from Dehydrated Product

Difco OGYE Agar Base

Aspergillus niger Escherichia coli Saccharomyces cerevisiae Saccharomyces cerevisiae

16404 25922 9763 9080

102-103 103-2103 102-103 102-103

Good Inhibition Good Good

1. Suspend 37 g of the powder in 1 L of purified water. Mix thoroughly. 2. Heat with frequent agitation and boil for 1 minute to completely dissolve the powder. 3. Autoclave at 121C for 15 minutes. 4. Aseptically add 10 mL of rehydrated Antimicrobic Vial Oxytetracycline to the medium at 50C. Mix well. 5. Test samples of the finished product for performance using stable, typical control cultures.
Difco Antimicrobic Vial Oxytetracycline

Expected Results
Refer to appropriate references and procedures for results.

References
1. Frank, Christen and Bullerman. 1993. In Marshall (ed.), Standard methods for the microbiological examination of dairy products, 16th ed. American Public Health Association, Washington, D.C. 2. Beuchat and Cousin. 2001. In Downes and Ito (ed.), Compendium of methods for the microbiological examination of foods, 4th ed. American Public Health Association, Washington, D.C. 3. Mossel, Visser and Mengerink. 1962. Lab. Pract. 11:109. 4. Mossel, Kleynen-Semmeling, Vincentie, Beerens and Catsaras. 1970. J. Appl. Bacteriol. 33:454.

Availability
Difco OGYE Agar Base
COMPF ISO SMD

1. Aseptically add 10 mL sterile purified water to the vial. 2. Shake to dissolve the contents.

Cat. No.

218111

Dehydrated 500 g

Difco Antimicrobic Vial Oxytetracycline

COMPF ISO SMD

Procedure
See appropriate references for specific procedures.

Cat. No.

232671

Vial 10 mL*

*Store at 2-8C.

Oatmeal Agar
Intended Use
Oatmeal Agar is used for cultivating fungi, particularly for macrospore formation. spores are called macroconidia, macroaleuriospores or macrospores and are produced by aerial sporulation.2 The detection of fungi is a great concern in the pharmaceutical, food and cosmetic industry.

Summary and Explanation

Fungi are extremely successful organisms, as evidenced by their ubiquity in nature. Of the estimated 250,000 species, fewer than 150 are known primary pathogens of humans.1 Identification and classification of fungi is primarily based on the morphologic differences in their reproductive structures.2 Fungi reproduce by producing spores.2 Large, multi-celled 412

Principles of the Procedure

Oatmeal is a source of nitrogen, carbon, protein and nutrients. Agar is the solidifying agent.