5 PREPARATION

• Dissolve 23.0 g of dehydrated media (BK009) in 1010 mL of distilled or ^ Reconstitution:

demineralized water. 23.0 g/1010mL
• Slowly bring to boiling, stirring until cx>mplete dissolution.
• Continué boiling for 2 minutes. / Sterilization:
• Do not autoclave. Bring to a boil for 2
• Cool rapidly to room temperatura. minutes
• Dispense into sterile tubes at 10 mL or in sterile viais at 100 mL.
r
Notes:
If overheated, a brick red selenium precipítate may form, denaturing the médium, which should be discarded.
Other volumes can be used depending on the protocol being followed.

6 INSTRUCTIONS FOR U S E

Selectiva enrichment for Salmonella in milk and other products (ISO 6579-1/A1) '^Inoculation:
10 mL
. Transfer 10 mL of pre-enrichment broth into 100 mL of Selenite Cystine broth.
Incúbate between 34 and 38°C fort 2 successive periods of 18 to 24 hours. Incubation:
. ísolate onto solid media; Brilliant Green agar and a second media of cholee, with 2 periods of 18 to 24 h
a loop of selectiva broth. :,j ,. . ; • ^ . ^ . i , between 34 and 38°C

Primaryenricliment for Sa/mone//ainsludge (FD/CEN/TR 15215-2)

'^Inoculation:
Transfer 10 mL of the sample suspensión into 90 mL of Selenite Cystine broth. 10 mL, 1 mL and 0.1 mL
. Transfer 1 mL and 0.1 mL of the sample suspensión into tubes of 10 mL of
Selenite Cystine broth. ^ Incubation:
. Incúbate at 36 ± 1 °C for 22 + 2 hours. 22 h at 36 °C
. Proceed with secondary enrichment by transferring 0,1 mL of the culture into
tubes of Rappaport Vassiliadis broth. ^ .
r
Notes:
For other uses, refer to the analytical protocols in vigor.
Beyond 24 hours of incubation, there is a decreased inhibitor effect in the media. A risk exists that a powerful
competitor like Proteus can grow at this time, which will destroy any Salmonella still present. On the other hand, a 48
hours incubation is required for the selective enrichment of Salmonella Pullorum.

7 QuALiTY CONTROL

Deliydrated media: cream-white powder, free-flowing and homogeneous.

Prepared media: light amber to pinkish solution, may contain a very slight precipítate.

Typicat culture response after 24 hours of incubation at 37 "C, followed by subcultures:

Microorganisms Growth
Salmonella Typhimurium WDCM 00031
> 10 characteristic colonies
+ Escheríchia coli WDCM 00013
+ Pseudomonas aeruginosa WDCM 00025

Salmonella Enteritidis WDCM 00030

> 10 characteristic colonies
+ Escheríchia coli WDCM 00013
+ Pseudomonas aeruginosa WDCM 00025
< 10 colonies
Enterococcus faecalis WDCM 00087
< 102 colonies
Escheríchia coli WDCM 00013

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 8 S T O R A G E / SHELF LIFE

Dehydrated base media: 2-20 °C.

The expiration date is indicated on the labe!.
Prepared media in tubes or vials ( * ) : 8 days at 2-8 °C, shielded from light and after verification of the absence of a
red precipítate.
(*) Benchmark valué determined under standard preparation conditions, following manufacturer's instructions.

9 PACKAGING

Dehydrated media:
500 a bottie BK009HA

10 BiBLIOGRAPHr

Leifson, E.. 1936. New selenite enrichment media for the isolation of typhoid and paratyphoid (Salmonella) bacilli.
American Journal of Hygiene, 24 : 423-432.
FD/CEN/TR 15215-2. Avril 2006. Caracterisation des boues. Detection et denombrement de Salmonella spp. dans les
boues, les sois, les amendements du sol, les supports de culture et biodéchets. Partie 2 : Méthode par
^nrichissement en milieu liquide sélénite-cystine puis en milieu de Rapport-Vassiliadis pour la détermination semi-
^antitative par la méthode du Nombre le Plus Probable (NPP).
NF U47-101. Novembre 2007. Methodes d'anaiyse en santé animale. Isolement et identification de tout sérovar ou de
sérovar(s) spécifié(s) de salmonelles chez les oiseaux.
NF U47-102. Janvier 2008. Methodes d'anaiyse en santé animale. isolement et identification de tout sérovar ou de
sérovar(s) spécifié(s) de salmonelles chez les mammiféres.
NF EN ISO 19250. Juin 2013. Quaiité de l'eau. Recherche de Salmonella spp.
NF EN ISO 6579-1. Avril 2017. Microbioiogie de la chaíne alimentaire - Méthode horizontale pour la recherche, le
denombrement et le sérotypage des Salmonella - Partie 1 : recherche des Salmonella spp.
NF EN ISO 6579-1/Al. March 2020. Miaobioiogy of the food chain - Horizontal method for the detection, enumeration
and serotyping of Salmonella - Parí 1 : detection of Salmonella spp. - Amendment 1 Broader range of incubation
temperaturas, amendment to the status of Annex D, and correction of the composition of MSRV and S C

11 ADDITIONAL INFORMATION

The Information provided on the iabeis take precedence over the formulations or instructions desaibed in this
/ ^ c u m e n t and are susceptible to modification at any time, without warning.

Document coda SELENITE CYSTINE BR0TH_ENv11

Creation date 04-2003
Updated 06-2020
Origin of revisión Update according to the standard NF EN ISO 6579-1/Al

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 ar
diaanostics
CERTIFICATE OF ANALYSIS

Product: S E L E N I T E CYSTINE BROTH 500g

Reference: BK0 0 9HA Expirydate: 2024.04.07
BatchN": 0037 655

Analysís Speciilcations Results

Characters

Appearance o f dehydrated F r e e - f l o w i n g and Conform

médium homogeneous powder
C^^r o f d e h y d r a t e d médium W h i t e creara Conform
aarance o f prepared S o l u t i o n may c o n t a i n a Conform
médium l i g h t precipítate
C o l o r o f p r e p a r e d médium Palé amber w i t h a p i n k r i n g Conform
on t h e s u r f a c e

Tests

pH o f p r e p a r e d médium 6, 8 t o 7,2 7,2

Microbiology

C u l t u r a l r e s p o n s e a f t e r 24 h o u r s o f i n c u b a t i o n a t 37°C, f o l l o w e d b y
subculture onto XLD Agar (NF E N I S O 1 1 1 3 3 , F D T 9 0 - 4 6 1 )

Growth Salmonella > 10 c h a r a c t e r i s t i c Conform

T y p h i m u r i u m ATCC 14028 colonies
(WDCM 0 0 0 3 1 ) + Escheríchia
c o l i ATCC 2 5 922 (WDCM
^A9v013) +
iudomonas a e r u g i n o s a ATCC
i; /853 (WDCM 0 0 0 2 5 )
Growth Salmonella > 10 c h a r a c t e r i s t i c Conform
E n t e r i t i d i s C I P 82.97 (WDCM colonies
0 0 0 3 0 ) + Escheríchia c o l i
ATCC 25922 (WDCM 0 0 0 1 3 ) +
Pseudomonas a e r u g i n o s a ATCC
2 7 8 5 3 (WDCM 0 0 0 2 5 )
Growth E n t e r o c o c c u s < 100 colonies Conform
f a e c a l i s ATCC 29212 (WDCM
00087)
G r o w t h Escheríchia c o l i < 100 c o l o n i e s Conform
ATCC25922 (WDCM 0 0 0 1 3 )

Beauvais July 1 8 , 2019

CONFORM Quality Control : Paséale GADONNA-WIDEHEM

T h i s certifícate i s e l e c t r o n i c a l l y generated ar.d r e q u l r e s no s l g n a t u r e
Biokar Diagnosücs - Rué des Quarante Mines - Z A C de Ther - Allonne - BPin245 - F6n002 B E A U V A I S C E D E X - F R A N C E
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