RAPPAPORT-VASSILIADIS R10 BROTH

Cat. no. K167 Rappaport-Vassiliadis R10 Broth, 16x125mm Tube, 10ml 20 tubes/box

INTENDED USE

Hardy Diagnostics Rappaport-Vassiliadis R10 Broth is recommended for the selective enrichment of Salmonella spp. from food and sewage polluted water.

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) A high concentration of magnesium chloride was included to inhibit growth of Proteus and Escherichia coli . The addition of malachite green inhibits coliforms. In addition, the low pH inhibits microorganisms other than Salmonella . In 1976, Vassiliadis, et al., described a modification of Rappaport Medium called R10. This formula featured a reduced amount of malachite green. (2) Hardy Diagnostics Rappaport-Vassiliadis R10 Broth is based on this formula.

FORMULA

Ingredients per liter of deionized water:*

Magnesium Chloride 13.4gm
Sodium Chloride 7.2gm
Pancreatic Digest of Casein 4.54gm
Potassium Phosphate 1.45gm
Malachite Green 36.0mg

Final pH 5.1 +/- 0.2 at 25ºC.

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

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC. away from direct light. Media should not be used if there are any signs of contamination, deterioration, discoloration, or if the expiration date has passed.Product is light and temperature sensitive; protect from light, excessive heat, and freezing.

PRECAUTIONS

PROCEDURE

Consult listed references for information regarding collection of specimens suspected of containing Salmonella . (3,5,6,8-10)

Pre-Enrichment
Inoculate the sample to be tested into Buffered Peptone Water or Lactose Broth and incubate for 18 to 24 hours at 35 to 37ºC.

Selective Enrichment
Inoculate the Rappaport-Vassiliadis R10 Broth with 0.1ml of the pre-enrichment culture. Inoculate a tube of Tetrathionate Broth in the same manner. Incubate The Rappaport-Vassiliadis R10 Broth at 41.5ºC for 48 hours. The Tetrathionate Broth is incubated for 48 hours at 42ºC.

Subculture
Subculture each enrichment broth to Brilliant Green Agar and XLD Agar and incubate at 37ºC. for 48 hours. After incubation, examine plates for colonies typical of Salmonella . Suspect Salmonella colonies should be identified by biochemical and serological tests. See listed references. (3,5,6,8-10)

For more detailed procedural information, consult listed references. (3,5,6,8-10)

INTERPRETATION OF RESULTS

Consult listed references for information on interpretation of growth in Rappaport-Vassiliadis R10 Broth. (3,5,6,8-10)

LIMITATIONS

Rappaport-Vassiliadis R10 Broth is a selective enrichment for Salmonella . Other biochemical and/or serological tests must be performed for complete identification. See listed references. (3,5,6,8-10)

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, slides, staining supplies, other culture media, microscopes, incinerators, and incubators, etc., as well as other biochemical and serological reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Salmonella enterica
ATCC® 14028
A 18-24hr 35°C Aerobic Growth when subcultured to XLD Agar
Escherichia coli
ATCC® 25922
B 18-24hr 35°C Aerobic Inhibited when subcultured to XLD Agar
Proteus vulgaris
ATCC® 8427
B 18-24hr 35°C Aerobic Inhibited when subcultured to XLD Agar

User Quality Control

PHYSICAL APPEARANCE

Rappaport-Vassiliadis R10 Broth should appear clear, and blue-green in color.

REFERENCES

1. Rappaport, F., et al. 1956. J. Clin. Path. ; 9:261-266.

2. Vassiliadis, P., et al. 1978. J. Appl. Bact. ; 44:233-239.

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

4. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology , C.V. Mosby Company, St. Louis, MO.

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

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

7. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria , Vol. I. Williams & Wilkins, Baltimore, MD.

8. FDA. 1995. Bacteriological Analytical Manual , 8th ed. FDA.

9. Marshall, R.T., ed. 1992. Standard Methods for the Examination of Dairy Products , 16th ed. APHA, Washington, D.C.

10. Vanderzant, C. and D.F. Splittstoesser, (ed.). 1992. Compendium of Methods for the Microbiological Examination of Foods, 3rd ed. APHA, Washington, D.C.

11. Greenberg, A.E., et al., (ed.). 1992. Standard Methods for the Examination of Water and Wastewater, 18th ed. APHA, Washington, D.C.

12. U.S. Pharmacopeia , 22nd rev. 1990. U.S. Pharmacopeial Convention, Rockville, MD.

13. Quality Assurance for Commercially Prepared Microbiological Culture Media , M22-A2, Vol. 16, No. 16. 1996. Clinical Laboratory Standards Institute (CLSI - formerly NCCLS), Villanova, PA.


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

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