rappaport-vassiliadis soja broth (rvs)

RAPPAPORT-VASSILIADIS SOJA BROTH (RVS)
INTENDED USE
Rappaport-Vassiliadis Soja Broth is used for the selective enrichment of Salmonella in milk, dairy
products, other food products, water and in the context of animal health analyses in mammals.
HISTORY
The composition of the medium was developed by Rappaport following the observation that
Salmonella was more resistant to hypertonic media than most other enterobacteria. In his
experiments, Rappaport showed that magnesium chloride was the most effective of all the salts
tested. Selectivity of the medium was increased still further by the addition of malachite green.
Vassiliadis subsequently showed that a larger number of Salmonella could be recovered by reducing
the malachite green content and incubating at 43°C instead of 37°C. Subsequent studies by Peterz et
al. showed the effects of incubation temperature and magnesium chloride concentration on the
recovery performance of the medium. Finally, van Schothorst and Renaud modified RappaportVassiliadis Broth by replacing the casein peptone with a soy peptone and by incorporating a
potassium hydrogenophosphate buffer into the formula, resulting in a greater stability of the medium
over time.
PRINCIPLES
- The high concentration of magnesium chloride and the presence of malachite green reduce the
growth of bacteria other than salmonellae.
- Growth of Salmonella Typhi and Shigella is inhibited by malachite green.
- The use of Selenite broth in parallel to Rappaport-Vassiliadis Soja Broth (RVS) is recommended
for samples contaminated by Salmonella Typhi, Salmonella Cholerasuis and other salmonella
serotypes.
PREPARATION
-
Dissolve 26.6 g of dehydrated medium (BK148) in 1 liter of distilled or deionized water.
Stir slowly until complete dissolution.
Dispense in tubes or flasks.
Sterilize in an autoclave at 115°C for 15 minutes.
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INSTRUCTIONS FOR USE
- Cool the medium to 25°C.
- Transfer 0.1 mL of inoculum into a tube of 10 mL of medium prepared as described or into a tube
of ready-to-use media BM074 (respect the 1:100 ratio). Inoculum generally constitutes a
preenrichment medium : Lactose Broth (BK082) or Buffered Peptone Water (BK018, BK131,
BM010 or BM057).
- Incubate for 18-24 hours at 41.5°C.
- Carry out an isolation in several selective media for Salmonella, with a platinum loop.
TYPICAL COMPOSITION
(can be adjusted to obtain optimal performance)
For 1 liter of medium :
- Papaic digest of soybean meal ......................................................4.50 g
- Sodium chloride .............................................................................7.20 g
- Potassium dihydrogen phosphate..................................................1.26 g
- Dipotassium phosphate .................................................................0.18 g
- Magnesium chloride anhydrous .....................................................13.4 g
- Malachite green oxalate ..............................................................36.0 mg
pH of the ready-to-use medium at 25°C : 5.2 ± 0.2.
QUALITY CONTROL
- Dehydrated medium : bluish powder, free-flowing and homogeneous.
- Prepared medium : blue solution, limpid.
- Typical culture response after 24 hours of incubation at 41.5°C, followed by subculture in XLD
Agar (ISO 6579) :
Subculture on XLD Agar
(ISO 6579)
Microorganisms
Salmonella Typhimurium
+ Escherichia coli
+ Pseudomonas aeruginosa
ATCC® 14028
ATCC 25922
ATCC 27853
≥ 10 characteristic colonies
(red with black center)
Salmonella Enteritidis
+ Escherichia coli
+ Pseudomonas aeruginosa
CIP 82.97
ATCC 8739
ATCC 27853
≥ 10 characteristic colonies
(red with black center)
Enterococcus faecalis
Escherichia coli
ATCC 29212
ATCC 25922
≤ 10 colonies
≤ 100 colonies
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Biokar Diagnostics – Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 – F60002 Beauvais Cedex – France
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 – www.biokar-diagnostics.com
STORAGE / SHELF LIFE
Dehydrated medium : 2-30°C, shielded from light.
- The expiration date is indicated on the label.
Prepared medium (benchmark value*) :
- Media in tubes : 6 months at 2-8°C, shielded from light.
Ready-to-use media in tubes :
- Store between 2-8°C, shielded from light.
- The expiration date is indicated on the label.
Code
PACKAGING
Ready-to-use media in tubes :
- 50 x 10 mL
BM07408
Dehydrated medium :
- 500 g bottle
BK148HA
BIBLIOGRAPHY
RAPPAPORT, F., KONFORTI, N., and NAVON, B.. 1956. A new enrichment medium for certain salmonellae. Journal of
Clinical Pathology, 9 : 261-266.
VASSILIADIS, P., KALAPOTHAKI, V., TRICHOPOULOS, D., PAPADAKIS, J.A., and SERIE, C.H.. 1979. Recent experience
on the use of modified Rappaport’s medium (R 10/43°C) for the isolation of Salmonella. Quality assurance and quality control
of microbiological culture media. Dr Janet E.L. Corry. London, 141-145.
van SCHOTHORST, M., and RENAND, A.M.. 1983. Dynamics of Salmonella isolation with modified Rappaport medium (R
10). Journal of Applied Bacteriology, 54 , 209-215.
van SCHOTHORST, M., RENAUD, A., and van BEEK, C.. 1987. Salmonella isolation using RVS broth and MLCB agar.
Food Microbiology, 4 , 11-18.
PETERZ, M., WIBERG, C., and NORBERG, P. 1989. The effect of incubation temperature and magnesium chloride
concentration on growth of salmonella in home-made and in commercially available dehydrated Rappaport-Vassiliadis
broths. Journal of Applied Bacteriology, 66 : 523-528.
ISO 6340. Décembre 1995. Qualité de l’eau. Recherche de Salmonella.
ISO 6785 / IDF 93. Mai 2001. Lait et produits laitiers. Recherche de Salmonella spp.
XP CEN ISO/TS 11133-2 (V 08-104-2). Janvier 2004. Microbiologie des aliments. Guide pour la préparation et la production
des milieux de culture. Partie 2 : Guide général pour les essais de performance des milieux de culture.
NF EN ISO 6579 (V 08-013). Mars 2007. Microbiologie des aliments. Méthode horizontale pour la recherche des Salmonella
spp.
NF U 47-102. Janvier 2008. Méthodes d’analyse en santé animale. Isolement et identification de tout sérovar ou de
sérovar(s) spécifié(s) de salmonelles chez les mammifères.
Pr NF EN ISO 19250 (T 90-435). Mars 2008. Qualité de l’eau. Dosage d’espèces de Salmonella.
NF EN ISO 6785 (V 04-031). Avril 2008. Lait et produits laitiers. Recherche de Salmonella spp.
*Benchmark value refers to the expected shelf life when prepared under standard laboratory conditions following manufacturer’s instructions. It is provided as a
guide only and no warranty implied or otherwise is associated with this information.
The information provided on the package take precedence over the formulations or instructions described in this document.
The information and specifications contained in this technical data sheet date from 2009-02-17.
They are susceptible to modification at any time, without warning.
Code document : BK148/A/2003-01 : 5.
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Biokar Diagnostics – Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 – F60002 Beauvais Cedex – France
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 – www.biokar-diagnostics.com