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BBL Standard Methods Agar (APHA)


(Tryptone Glucose Yeast Agar)
L007506 Rev. 08 October 2006
QUALITY CONTROL PROCEDURES

INTRODUCTION
Standard Methods Agar is a standardized medium for the enumeration of microorganisms from materials of sanitary
importance.

II

PERFORMANCE TEST PROCEDURE


1. Liquefy the medium in the pour tube by heating in boiling water. Cool to 4550C and pour into sterile Petri dishes. Allow
to solidify for a minimum of 30 min.
2. Streak-inoculate the plates with a 0.01 mL calibrated loop using 10-1 dilutions of 18- to 24-h Trypticase Soy Broth cultures.
3. Incubate plates at 35 2C in an aerobic atmosphere.
4. Examine after 24, 48, and 72 h for growth.
5. Expected Results
Organisms
*Lactobacillus rhamnosus
*Staphylococcus aureus
Streptococcus pyogenes
Pseudomonas aeruginosa

ATCC
7469
25923
19615
27853

Recovery
Growth
Growth
Growth
Growth

*Recommended organism strain for User Quality Control.

III

ADDITIONAL QUALITY CONTROL


1. Examine tubes as described under "Product Deterioration."
2. Visually examine representative tubes to assure that any existing physical defects will not interfere with use.
3. Determine the pH potentiometrically at room temperature for adherence to the specification of 7.0 0.2.
4. Incubate uninoculated representative tubes aerobically at 2025C and 3035C and examine for 7 days for microbial
contamination.

IV

INTENDED USE
Standard Methods Agar (Plate Count Agar; Tryptone Glucose Yeast Agar) is used for obtaining microbial plate counts from milk
and dairy products, foods, water and other materials of sanitary importance.

SUMMARY AND EXPLANATION


Standard Methods Agar is made according to the American Public Health Association (APHA) formulation for Plate Count
Agar.1 Standard Methods Agar is recommended for obtaining plate counts for milk and other dairy products and may also be
used to determine the sanitary quality of foods, water and other materials.1-5
Each lot of BBL Standard Methods Agar base is subjected to the APHA quality control test and has met the APHA requirements.1,6
Appropriate references should be consulted for standard plate count procedures recommended by the APHA and other agencies.1-5

VI

PRINCIPLES OF THE PROCEDURE


Enzymatic digest of casein provides the amino acids and other complex nitrogenous substances necessary to support bacterial
growth. Yeast extract primarily supplies the B-complex vitamins, and dextrose is an energy source.

VII

REAGENTS

PRODUCT INFORMATION

Standard Methods Agar


Approximate Formula* Per Liter Purified Water
Pancreatic Digest of Casein ....................................................5.0
Yeast Extract ............................................................................2.5
Dextrose ..................................................................................1.0
Agar ......................................................................................15.0

g
g
g
g

*Adjusted and/or supplemented as required to meet performance criteria.

Warnings and Precautions: For Laboratory Use.


Tubes with tight caps should be opened carefully to avoid injury due to breakage of glass.
Observe aseptic techniques and established precautions against microbiological hazards throughout all procedures. After use,
prepared tubes, specimen containers and other contaminated materials must be sterilized by autoclaving before discarding.
Storage Instructions: On receipt, store tubes in the dark at 225C. Avoid freezing and overheating. Do not open until ready to
use. Minimize exposure to light. Tubed media stored as labeled until just prior to use may be inoculated up to the expiration date
and incubated for the recommended incubation times. Allow the medium to warm to room temperature before inoculation.
Product Deterioration: Do not use tubes if they show evidence of microbial contamination, discoloration, drying or other signs
of deterioration.
VIII SPECIMEN COLLECTION AND HANDLING
Consult appropriate texts for detailed information and recommended procedures.1-5

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IX

PROCEDURE
Material Provided: Standard Methods Agar
Materials Required But Not Provided: Ancillary culture media, reagents, quality control organisms and laboratory equipment as
required.
Test Procedure: Observe aseptic techniques.
Consult appropriate references for information regarding the processing and inoculation of food, water samples and other
materials.1-5
Usually 1-mL samples of appropriate dilutions of the test sample are pipetted into sterile Petri dishes and molten, cooled
Standard Methods Agar is added followed by gently mixing to distribute the sample dilution throughout the agar. Incubate
hardened plates for 48 3 h at 32 1C (dairy products) or 35 0.5C (foods) in an aerobic atmosphere.
User Quality Control: See "Quality Control Procedures."
A single electrode of sufficiently small size to fit into the tubes should be used to determine the pH potentiometrically of
tubed media. The tip of the electrode should be placed below the surface of broth media.

RESULTS
Follow recommended procedures for the counting of colonies and the reporting of results.1-5

XI

LIMITATIONS OF THE PROCEDURE


For identification, organisms must be in pure culture. Morphological, biochemical, and/or serological tests should be performed
for final identification. Consult appropriate texts for detailed information and recommended procedures.1-6

XII

AVAILABILITY
Cat. No.
221030
297943

Description
BBL Standard Methods Agar Deeps (Pour Tubes), 20 mL, Pkg. of 10 size A tubes
BBL Standard Methods Agar Deeps (Pour Tubes), 20 mL, Ctn. of 100 size A tubes

XIII REFERENCES
1. Wehr, H.M. and J.H. Frank (ed.). 2004. Standard methods for the examination of dairy products, 17th ed. American Public Health Association,
Washington, D.C.
2. Downes, F.P. and K. Ito (ed.). 2001. Compendium of methods for the microbiological examination of foods, 4th ed. American Public Health
Association, Washington, D.C.
3. Clesceri, L.S., A.E. Greenberg, and A.D. Eaton (ed.). 1998. Standard methods for the examination of water and wastewater, 20th ed. American
Public Health Association, Washington, D.C.
4. Horwitz, W. (ed.). 2000. Official methods of analysis of the AOAC International, 17th ed. AOAC International, Gaithersburg, Md.
5. U.S. Food and Drug Administration. 2002. Bacteriological analytical manual, (on line). AOAC International, Gaithersburg, Md.
http://www.cfsan.fda.gov/~ebam/bam-toc.html.
6. Marth, E.H. (ed.). 1978. Standard methods for the examination of dairy products, 14th ed. American Public Health Association, Washington, D.C.

Becton, Dickinson and Company


7 Loveton Circle
Sparks, Maryland 21152 USA
800-638-8663
ATCC is a trademark of the American Type Culture Collection.
BD, BD Logo, BBL, and Trypticase are trademarks of Becton, Dickinson and Company. 2006 BD.

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