CRITERION™ RAPPAPORT-VASSILIADIS BROTH, USP

Cat. no. C8780 CRITERION™ Rappaport-Vassiliadis Broth, USP 54.8gm
Cat. no. C8781 CRITERION™ Rappaport-Vassiliadis Broth, USP 500gm
Cat. no. C8782 CRITERION™ Rappaport-Vassiliadis Broth, USP 2kg
Cat. no. C8783 CRITERION™ Rappaport-Vassiliadis Broth, USP 10kg
Cat. no. C8784 CRITERION™ Rappaport-Vassiliadis Broth, USP 50kg

INTENDED USE

Hardy Diagnostics CRITERION™ Rappaport-Vassiliadis Broth, USP is recommended for the selective enrichment of Salmonella spp. from food samples and conforms to the Harmonized USP/EP/JP requirements.

This dehydrated culture medium is a raw material intended to be used in the making of prepared media products, which will require further processing, additional ingredients, or supplements.

SUMMARY

Rappaport Medium was initially developed by Rappaport et al. in 1956 as an alternative to Tetrathionate Broth for the enrichment of Salmonella.(1) This formulation features magnesium chloride to inhibit Proteus spp. and Escherichia coli; malachite green to inhibit coliforms, and a high osmotic pressure and/or low pH to inhibit accompanying microbial flora other than Salmonella. In 1976, Vassiliadis et al. described a modification of Rappaport Medium called R10.(2) This formula features a reduced concentration of malachite green and an increased incubation temperature. It was later shown in 1989 by Peterz et al. that incubation at 41.5 +/-0.5ºC. for 24 hours significantly improved the recovery of Salmonella spp.(3)

Rappaport-Vassiliadis Broth, USP is a modification of Rappaport-Vassiliadis R10 Broth and uses soy peptone as the nitrogen and vitamin source. Studies show that soy peptone enhances the growth of Salmonella spp. and counteracts the risk of potential Bovine Spongiform Encephalopathy (BSE) exposure associated with bovine derived products. Rappaport-Vassiliadis Broth, USP conforms to the Harmonized United States Pharmacopoeia (USP), European Pharmacopoeia (EU), and Japanese Pharmacopoeia (JP).(4-7) The medium selectively enriches for Salmonella spp., although malachite green may inhibit the growth of more sensitive strains of Salmonella, such as S. typhi and S. choleraesuis.

FORMULA*

Gram weight per liter: 27.4gm/L
Magnesium Chloride, Anhydrous 13.4gm
Sodium Chloride 8.0gm
Soy Peptone 4.5gm
Dipotassium Phosphate 1.26gm
Monopotassium Phosphate 0.18gm
Malachite Green 0.036gm

Final pH 5.2 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented to meet performance criteria.

NOTE: Since magnesium chloride hexahydrate contains too much water to be effectively used in the manufacture of dehydrated culture media, magnesium chloride anhydrous is used as an acceptable alternative. The actual overall concentration of magnesium chloride is equivalent with this substitution; yet, the adjustment requires additional modifications to the USP formulation to correct for molecular weight. None of these modifications affect the overall performance or intended use of this item as indicated on the Certificate of Analysis, which shows harmonized growth promotion criteria are met per USP.(4-7)

STORAGE AND SHELF LIFE

Store the sealed bottle(s) containing dehydrated culture medium at 2-30ºC. Dehydrated culture medium is very hygroscopic. Keep lid tightly sealed. Protect dehydrated culture media from moisture and light. The dehydrated culture media should be discarded if it is not free-flowing or if the color has changed from its original pale green.

Store the prepared culture media at 2-8ºC.

PRECAUTIONS

METHOD OF PREPARATION FOR DEHYDRATED CULTURE MEDIA

1. Suspend 27.4gm of the dehydrated culture media in one liter of distilled or deionized water. Stir to mix thoroughly.

2. Heat as necessary to dissolve completely.

3. Dispense 10ml into glass tubes and cap loosely.

4. Sterilize in the autoclave at 118ºC. for 12 minutes.

PROCEDURE AND INTERPRETATION OF RESULTS

For information on specific procedures and the interpretation of results, consult appropriate reference guidelines or refer to the prepared media Instructions for Use (IFU) for Cat. No. K246.

LIMITATIONS

Certain strains of Salmonella, such as S. typhi and S. choleraesuis, may be inhibited on this medium. Therefore, isolation techniques should include a variety of enrichment broths and selective media for best recovery.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as autoclaves, incinerators, and incubators, etc., are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Salmonella enterica
ATCC ® 14028**
J 18-24hr 35°C Aerobic Positive upon subculture to XLD
Staphylococcus aureus
ATCC ® 6538**
B 48hr 35°C Aerobic Inhibited upon subculture to XLD

** Recommended QC strains for User Quality Control according to the CLSI document M22 when applicable.

User Quality Control

Physical Appearance

CRITERION™ Rappaport-Vassiliadis Broth, USP powder should appear homogeneous, free-flowing, and pale green in color. The prepared media should appear clear and blue in color.

REFERENCES

1. Rappaport, F., N. Konforti, and B. Navon. 1956. A New Enrichment Medium for Certain Salmonellae. J. Clin. Pathol.; 9:261-266.

2. Vassiliadis, P., D. Trichoppoulos, A. Kalandidi, and E. Xirouchaki. 1978. Isolation of Salmonellae from Sewage with a New Procedure of Enrichment. J. Appl. Bacteriol.; 44:233-239.

3. Peterz, M. C. Wiberg, and P. Norberg. 1989. The Effect of Incubation Temperature and Magnesium Chloride Concentration on Growth of Salmonella in Homemade and Commercially Available Dehydrated Rappaport-Vassiliadis Broths. J. Appl. Bacteriol.; 66:523-528.

4. United States Pharmacopeial Convention. 2007. The United States Pharmacopeia, Amended Chapters 61, 62, 111. The United States Pharmacopeial Convention, Rockville, MD.

5. The Official Compendia of Standards. 2008. USP27-NF22. United States Pharmacopeial Convention, Rockville, MD.

6. Directorate for the Quality of Medicines of the Council of Europe (EDQM). 2007. The European Pharmacopoeia, Amended Chapters 2.6.12, 2.6.13, 5.1.4, Council of Europe, 67075 Strasbourg Cedex, France.

7. Japanese Pharmacopoeia. 2007. Society of Japanese Pharmacopoeia. Amended Chapters 35.1, 35.2, 7. The Minister of Health, Labor, and Welfare.

8. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

9. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

10. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.

11. American Public Health Association. Standard Methods for the Examination of Dairy Products, APHA, Washington, D.C.

12. APHA Technical Committee on Microbiological Methods for Foods. Compendium of Methods for the Microbiological Examination of Foods, APHA, Washington, D.C.

13. American Public Health Association. Standard Methods for the Examination of Water and Wastewater, APHA, Washington, D.C.

14. U.S. Food and Drug Administration. Bacteriological Analytical Manual. AOAC, Arlington, VA.
http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm2006949.htm.

15. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

16. Association of Official Analytical Chemists. Official Methods of Analysissm, AOAC, Washington, D.C.


ATCC is a registered trademark of the American Type Culture Collection.

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