Novobiocin Selective Supplement (20 mg)

Collect, dilute and prepare samples and volumes as required according to specifications, directives, official standard regulations and/or expected results.

Reconstitute the vial with 6 mL of sterile diluent, pre-warmed to approx. 37 °C, and add to 500 mL of Tetrathionate Broth Base cooled to room temperature.

Note: for Rappaport-Vassiliadis Medium add 1 vial per litre of medium.

For Salmonella enrichment use Tetrathionate Broth

Immediately before inoculation, supplement tubes or flasks to get the following concentrations:

Incubate the tubes tightly closed aerobically at 42 ± 2 °C for 24 h.

(Incubation times, temperature and sample volumes may vary depending on the sample, on the specifications, etc.)

Read the turbidity (growth indicator) and inoculate any confirmatory, secondary medium by streaking methodology or by spiral method, like XLD, BPLS, etc., to confirm results. After proper incubation, enumerate all the colonies that have appeared onto the surface of the secondary agar.

Presumptive isolation or recovery of Salmonella must be confirmed by further microbiological and biochemical tests.

Each laboratory must evaluate the results according to their specifications.

Reconstitute the original freeze-dried vial by adding 6 mL of sterile distilled water.

Note: each vial is sufficient to supplement 1 L of Rappaport-Vassiliadis Medium.

Physical/chemical control

Microbiological control

Reconstitute 1 vial as indicated in Composition; shake and dissolve completely.

Add 1 vial to 500 mL of medium base. DO NOT HEAT once supplemented.

Distribute the complete medium, cooled to 50 °C, into 90 mm plates.

Incubate according to the instructions for the complete medium indicated in Composition.

Sterility control

Study 5 vials - reconstitute and dissolve each one in 100 mL of TSA + neutralizers - pour into 90 mm plates.

Incubation 24 hours at 30-35 °C and 72 hours at 20-25 °C: NO GROWTH.

• EUROPEAN PHARMACOPOEIA (2005) 5th ed. § 2.6.13. Microbiological Examination of Non-Sterile Products (Test for Specified Microorganisms). EDQM. Council of Europe. Strasbourg.
• De SMEDT, J.M., R. BOLDERDJIK, H. RAPPOLD & D. LAUTENSCHLAEGER (1986) Rapid Salmonella detection in foods by motility enrichment on a Modified Semisolid Rappaport Vassiliadis Medium. J. Food Protect. 49:510-514.
• De SMEDT, J.M. & R. BOLDERJIK (1987) Dynamics of Salmonella Isolation with Modified Semisolid Rappaport Vassiliadis Medium. J. Food Protect. 50:658-661.
• HOLBROOCK, R., J.M. ANDERSON, A.C. BAIRD-PARKER, L.M. DODDS, D. SAWHNEY, S.H. STRUCHBURY & D. SWAINE (1989) Rapid detection of Salmonella in food: A convenient two-day procedure. Lett. Appl. Microbiol. 8:139-142.
• ISO Standard 6579-1 (2017) Microbiology of food chain - Horizontal method for the detection, enumeration and serotyping of Salmonella - Part 1: Detection of Salmonella spp.