Biolife Italiana Srl Technical Sheet
N °401981-rev.1 BE –1 07/2009 page 1/ 1
RAPPAPORT VASSILIADIS SOY (RVS) BROTH
Powdered and ready to use selective liquid medium
for the enrichment of Salmonella.
TYPICAL FORMULA (g/l)
Soy Peptone 4.500
Sodium Chloride 7.200
Potassium Dihydrogen Phosphate 1.260
Dipotassium Hydrogen Phosphate 0.180
Magnesium Chloride 13.400
Malachite Green Oxalate 0.036
DIRECTIONS
Suspend 26.6g in 1000ml of cold distilled water. Heat to dissolve, then distribute 10 ml into screw-cap
º
bottles or tubes and sterilise by autoclaving at 115 C for 15 minutes.
Final pH 5.2 ± 0.2
DESCRIPTION
Rappaport Vassiliadis Soy (RVS) Broth is used as a selective enrichment medium for the isolation of
Salmonella from food, water and environmental specimens. RVS Broth has the same formulation as
Rappaport Vassiliadis Medium, except for the addition of dipotassium hydrogen phosphate, and
reduced concentration levels of magnesium chloride and potassium dihydrogen phosphate.
The modified buffer salts concentration allows a better maintenance of pH and an improved isolation of
salmonellae. RVS Broth is based on the revised formula described by van Schothorst et al.
Rappaport Vassiliadis Soy (RVS) Broth is recommended by ISO 6579:2002 the horizontal method for
detection Salmonella spp. (including S.typhi) in foodstuffs, together with Mulller Kauffman
Tetrathionate Novobiocin Broth.
TECHNIQUE
1. The procedure recommended by ISO 6579:2002, is as follows:
2. Add 25g sample portion to 225ml of Buffered Peptone Water. If the required test portion is other
than 25g, use a suitable quantity of Buffered Peptone Water to yield approximately 1/10 dilution
(m/v).
3. Incubate the initial suspension at 37°C for a minimum of 16 hours and not more than 20 hours.
4. Transfer 0.1ml of the pre-enriched culture to a tube containing 10ml of Rappaport Vassiliadis Soy
(RVS) Broth and 1ml to a flask containing 10ml of Mueller Kauffmann Novobiocin Broth (MKTTn)
5. Incubate the inoculated RVS Broth at 41.5°C ± 1°C for 24hrs ± 3hrs.
6. Incubate the inoculated MKTTn at 37°C ± 1 for 24hrs ± 3.
7. Using a culture obtained from the RVS Broth inoculate by means of a 3mm loop, a large-size Petri
dish or two 90mm Petri dishes containing XLD Medium (REF 402206), proceed in the same way
from the enrichment tube by inoculating a second plating medium (e.g. Chromogenic Salmonella
Agar (REF 405350), or another suitable selective Salmonella plating-out medium chosen by the
laboratory).
8. Using the cultures obtained in MKTTn after 24 hours of incubation, repeat the procedure with the
same two selective plating-out media.
9. Invert the dishes and incubate at 37°C for 24hrs. ± 3 hrs.
10. Examine for the presence of typical colonies. For confirmation take from each dish of each
selective medium at least one typical or suspected colony and a further 4 colonies if the first is
negative. Streak the selected colonies onto the surface of Nutrient Agar and incubate at 37°C for
24hrs. Use pure cultures for biochemical and serological confirmation. Biochemical confirmation
tests include: TSI Agar, Urea Agar, L-Lysine Decarboxylase Medium, detection of -galactosidase,
VP reaction, indole detection. Serological confirmation includes the detection of the presence of
Salmonella O-, Vi- and H antigens by slide agglutination test. Biochemical confirmation can be
substituted with the rapid test MUCAP (REF 191500). All the colonies MUCAP positive must be
serologically confirmed.
Biolife Italiana S.r.l. Viale Monza 272, 20128 Milan, Italy. Tel. n° ++39 (0)2 25 209 1, Fax n° ++39 (0)2 25 76 428
E-mail: [email protected] ; Web: www.biolifeitaliana.it
Biolife Italiana Srl Technical Sheet
N °401981-rev.1 BE –1 07/2009 page 2/ 2
USER QUALITY ASSURANCE (42°C-24hrs, subculture on TSA)
Productivity control
S.typhimurium ATCC 14028: growth
Selectivity control
E.coli ATCC 25922: partially or completely inhibited
STORAGE
Dehydrated medium: 10-30°C
User prepared tubes: 3 months at 2-8°C
REFERENCES
• ISO 6579 Microbiology of food and animal foodstuff - Horizontal method for the detection of
Salmonella spp. 2002.
• Van Schothorsts, M., Renaud, A., van Beek, C. (1987) Food Microbiology 4, 11-18
PACKAGING
4019812 Rappaport Vassiliadis Soy (RVS) Broth, 500g (18.7 L)
4019814 Rappaport Vassiliadis Soy (RVS) Broth, 5 kg (187 L)
551981 Rappaport Vassiliadis Soy (RVS) Broth, 20 x 10 ml ready to use tubes
5119812 Rappaport Vassiliadis Soy (RVS) Broth, 6 x 100 ml ready to use flasks
Biolife Italiana S.r.l. Viale Monza 272, 20128 Milan, Italy. Tel. n° ++39 (0)2 25 209 1, Fax n° ++39 (0)2 25 76 428
E-mail: [email protected] ; Web: www.biolifeitaliana.it
