RAPPAPORT-VASSILIADIS BROTH, USP

Cat. no. K246 Rappaport-Vassiliadis Broth, USP, 16x125mm Tube, 10ml 20 tubes/box

INTENDED USE

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

This product is not intended to be used for the diagnosis of human disease.

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

Ingredients per liter of deionized water:*

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 as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8ºC. away from direct light. Media should not be used if there are any signs of deterioration, discoloration, contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

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.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, applicator sticks, other culture media, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Salmonella enterica
ATCC ® 14028**
J*** 18-24hr 30-35°C Aerobic Growth
Staphylococcus aureus
ATCC ® 6538**
B 24hr 30-35°C Aerobic Inhibited

** Recommended QC strains for User Quality Control according to the CLSI document M22 and/or USP/EP, when applicable.

***Tested in accordance with USP <62>

USER QUALITY CONTROL

Physical Appearance

Rappaport-Vassiliadis Broth, USP 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. USP-NF . 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. 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. 2012. Official Methods of Analysis , 19th ed. AOAC, Washington, D.C.

17. The Official Compendia of Standards. USP General Chapter <62> Microbiological Examination of Nonsterile Products: Tests for Specified Microorganisms. USP-NF . United States Pharmacopeial Convention Inc., Rockville, MD.


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