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Dehydrated Culture Media

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BRILLIANCE SALMONELLA AGAR BASE

Code: CM1092

Brilliance™ Salmonella Agar (formerly OSCM II) is used to produce a selective medium for the presumptive identification and differentiation of Salmonella species.

Typical Formula*

gm/ litre

Inhibigen™ mix

14

Chromogenic Mix

25

Agar

15

pH 7.3 ± 0.1 @ 25ºC  
* Adjusted as required to meet performance standards
† pH of supplemented medium

SALMONELLA SELECTIVE SUPPLEMENT

Code: SR0194

Vial Contents (each vial is sufficient for 500ml of medium) per vial per litre
Novobiocin

2.5mg

5mg

Cefsulodin

6mg

12mg

Brilliance Salmonella Agar is the isolation medium used in the Oxoid Salmonella Precis rapid culture method. For information simply download the Brilliance Salmonella Agar data sheet (1MB) or the Salmonella Precis data sheet (1396KB) in PDF format. Please confirm prepared media codes with your local supplier.

Directions
Suspend 27g of Brilliance Salmonella Agar Base in 500ml of distilled water. Add the contents of 1 vial of Salmonella Selective Supplement, resuspended as directed. Mix well and sterilise by bringing to the boil with frequent agitation. Cool to around 50ºC, mix well and pour into sterile Petri dishes.
NOTE: It is critical that the selective supplement is added prior to heating. When preparing volumes greater than 1 litre contact Oxoid Technical Support for directions.

Description
An Inhibigen™ compound is comprised of two components, combined together by a bond that can only be cleaved by a specific enzyme. When bound together, the inhibitor compound is not toxic and therefore can exist in a medium without harming micro-organisms. Once inside the cell, the bond will be cleaved if the target enzyme is present. When the bond is cleaved, the inhibitor molecule is released and disrupts cell wall synthesis, causing death of the organism. As cells die and lyse, free inhibitor is released but cannot be taken up by other cells, resulting in targeted inhibition. The Inhibigen in Brilliance Salmonella Agar targets Escherichia coli. Novobiocin and cefsulodin, presented as a freeze-dried supplement (SR0194), are added to the medium to inhibit the growth of other competing flora such as Proteus spp. and Pseudomonas spp.

Differentiation of Salmonella from the other organisms that grow on Brilliance Salmonella Agar is achieved through the inclusion of two chromogens that also target specific enzymes: caprylate esterase and ß-glucosidase. Caprylate esterase is an enzyme present in all samonellae as well as some species of Klebsiella, Enterobacter and Proteus1. Organisms possessing caprylate esterase cleave the chromogen to release an insoluble purple chromophore. As the cells grow, the chromophore builds up and produces a purple-coloured colony. Some Enterobacteriaceae, including Klebsiella and Enterobacter but not Salmonella, possess ß-glucosidase2. If these organisms grow, they will form blue or dark blue colonies, even if they are esterase positive,which make them easy to differentiate from purple Salmonella colonies.

Technique

Testing food samples
Brilliance Salmonella Agar can be used for testing clinical (faecal) samples, either directly or following a suitable enrichment protocol. It can also be used for food and environmental samples following a variety of enrichment procedures e.g. ISO, NMKL, FDA, HPA, etc. Depending on the method, Brilliance Salmonella Agar can be streaked with sample or post-incubation enrichment broth. The plates are then incubated at 37ºC for 24 ± 3h. Salmonella produce purple colonies.

The following method is a summary of ISO 6579:2002 +A1:20073 illustrating how Brilliance Salmonella Agar can be used alongside XLD Medium, when following this method:

  1. Add 25g of food sample to 225ml of Buffered Peptone Water and stomach for a minimum of 30 seconds to mix the sample
  2. Incubate the broth at 37ºC for 18 ± 2h
  3. Gently agitate the bag then, using a pipette, inoculate 1ml into 10ml of MKTTn Broth and 0.1ml into 10ml of RVS Broth. Incubate MKTTn broth at 37ºC for 24 ± 3h and RVS broth at 41.5ºC for 24h ± 3h
  4. Using a microbiological loop, inoculate each broth onto one XLD Agar plate and one Brilliance Salmonella Agar plate to give a total of 4 plates.
  5. Incubate the plates at 37ºC for 24h ± 3h
  6. Confirm presumptive red colonies with black centres from XLD and purple colonies from Brilliance Salmonella Agar as Salmonella spp by appropriate biochemical and serological methods. Refer to ISO 6579:2002 +A1:20073.

The following Oxoid products may be useful to confirm a presumptive Salmonella identification: Nutrient Agar (CM0003), TSI Agar (CM0277), Urea Agar (CM0053 & SR0020K), Lysine Decarboxylase Medium (CM0308), Methyl-Red Voges-Proskauer (MRVP) Medium (CM0043), Tryptone Soya Broth (CM0129), Agglutinating Anti-sera (see Oxoid product list for codes); Oxoid Salmonella Latex Test Kit (DR1108), Microbact (see Oxoid product list for codes); O.B.I.S. Salmonella (ID0570). Contact your local Oxoid supplier for prepared media codes.

Testing clinical samples

Inoculate the plate directly with pea sized bead or loopful of specimen. Incubate plates at 37°C or for 24h±3h. Alternatively, pre-enrich in suitable selective broth prior to inoculation onto a Brilliance Salmonella plate. Use an incubation protocol appropriate to the broth chosen. Purple colonies should be confirmed as Salmonella spp by appropriate biochemical and serological methods .

For further instructions on the use and interpretation of Brilliance Salmonella Agar with clinical samples, simply download the data sheet (885KB) in PDF format. Please confirm prepared media codes with your local supplier.

Storage Conditions and Shelf Life
Brilliance Salmonella Agar should be stored in the tightly capped original container at 10-30ºC. Salmonella Selective Supplement should be stored in the dark at 2-8ºC. When stored as directed, the un-opened products will remain stable until the expiry date printed on the packaging.

Locally prepared Brilliance Salmonella Agar plates can be stored for up to 2 weeks when made from CM1092 and SR0194 according to the manufacturer’s instructions and stored at 2-8ºC, out of direct sunlight. A longer shelf life may be attainable, but should be validated under the relevant, local manufacturing and storage conditions.

Appearance
Dehydrated medium: White/cream coloured, free-flowing powder
Selective supplement: White, freeze-dried pellet
Prepared medium: White/cream coloured opaque gel

Quality control

Positive controls: Expected results
Salmonella Typhimurium ATCC®14028* Good growth; purple colonies
Klebsiella pneumoniae ATCC®13883* Good growth; blue colonies
Negative controls:  
Enterococcus faecalis ATCC®29212* Inhibited
Escherichia coli ATCC®25922* Inhibited
*This organism is available as a Culti-Loop®

Precautions
Brilliance Salmonella Agar is for in vitro diagnostic use only, by experienced microbiologists. It must not be used beyond the stated expiry date, or if the product shows any sign of deterioration.
Sterilise specimens, equipment and media properly after use.

Limitations
It should be noted that, as with all Salmonella media, organisms with atypical enzyme patterns may give anomalous reactions on Brilliance Salmonella Agar. A small number of atypical strains that may give a weak reaction or fail to grow, especially when low numbers are present in the sample. Brilliance Salmonella Agar is not recommended for the isolation of Salmonella typhi and Salmonella paratyphi. Please use Oxoid Salmonella Culture Medium (OSCM) CM1007 as your chromogenic medium for this purpose.

Refs
1. Cooke. V.M. et al, A Novel Chromogenic Ester Agar Medium for Detection of Salmonellae. Appl. Environ. Microbiol. Feb 1999 p. 807-812, Vol. 65, No. 2.
2. Garrity, G., Brenner, D.J., Krieg, N.R., Staley, J.R. (Eds.), Bergey’s Manual of Systematic Bacteriology. Vol 2, Part B. 2nd ed., 2005. US, Springer.
3. ISO 6579:2002 +A1:2007: Microbiology of food and animal feeding stuffs – Horizontal method for the detection of Salmonella spp.

ATCC® is a registered trademark of American Type Culture Collection

Brilliance Salmonella PDF (885KB)

 
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