XLD AGAR (ISO 6579)

INTENDED USE XLD (Xylose Lysine Desoxycholate) Agar is used to isolate Salmonella in food products, according to the standard NF EN ISO 6579, as well as Shigella, following detection protocol described in NF EN ISO 21567. XLD Agar is also used as a selective isolation medium for the detection of Salmonella in waters according to protocol described in the NF ISO 19250 standard.

HISTORY The medium was formulated by Taylor in order to increase the efficiency of recovery of pathogenic enterobacteria, particularly Shigella and other fastidious species which do not develop in other formulations containing highly toxic inhibitors.

PRINCIPLES - Sodium desoxycholate inhibits contaminating Gram-positive flora. - Xylose is fermented by practically all enteropathogenic bacteria, except for Shigella which are thus differentiated from the other species. After exhausting xylose, Salmonella decarboxylate lysine (via lysine decarboxylase) to cadaverine, causing the pH to rise. Colonies of salmonellae resemble those of shigellae in the medium having become basic. - The colonies formed are red in the presence of the indicator, phenol red. - The addition of lactose and sucrose to the medium enable coliform bacteria to decarboxylate lysine and thereby produce excess acidity, making the indicator turn yellow, favoring their differentiation. - In basic medium, pathogenic H2S-producers reduce ferric ammonium citrate and cause a blackening due to the production of iron sulfide at the center of the colonies. Non-pathogenic bacteria which do not decarboxylate lysine acidify the medium, a result of sugar fermentation. The pH decrease prevents the colonies from blackening.

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PREPARATION Suspend 52.9 g of dehydrated medium (BK168) in 1 liter of distilled or deionized water. Heat to about 90C with frequent agitation. Stop heating when the medium is completely dissolved. Do not autoclave.

NOTE : Excessive heating or prolonged holding at 44-47C may cause precipitation, so the colonies may furnish less clear-cut reactions. The medium should be clear and orange-red.

INSTRUCTIONS FOR USE Cool and maintain the medium at 44-47C. Pour into sterile Petri dishes. Let solidify on a cold surface. Dry in an incubator with the covers partially removed. Inoculate by streaking on the surface of the medium prepared as described (or use ready-to-use plates BM087), using enrichment media used for the detection of Salmonella. - In parallel, transfer the inoculum to another selective isolation medium such as nz agar (BK032 or BM083) or COMPASS Salmonella agar (BM066). Incubation for the detection of Salmonella in food samples: - Incubate at (37 2)C for 24 and 48 hours. Incubation for the detection of Salmonella in water samples: - Incubate at (36 2)C for (24 3) hours.

RESULTS Shigella, Providencia, Edwardsiella and Salmonella Paratyphi ferment xylose slowly or not at all, in contrast to other bacteria. Salmonella is distinguished from other genera by its ability to decarboxylate lysine. Hydrogen sulfide production in a basic medium leads to the formation of colonies with a black center, while the black color is inhibited in an acid medium. The colonies have the following appearance :

Characteristics Yellow colonies LDC (-), sometimes H2S (+) Red colonies LDC (+), H2S (-) Red colonies with black center LDC (+), H2S (+)

Microorganisms Escherichia coli, Citrobacter, Enterobacter, Proteus, Serratia, Klebsiella Shigella, Providencia, Salmonella Salmonella, Arizona, Edwardsiella

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TYPICAL COMPOSITION (can be adjusted to obtain optimal performance) For 1 liter of medium : - Yeast extract ....................................................................................3.0 g - L-Lysine ...........................................................................................5.0 g - Lactose ............................................................................................7.5 g - Sucrose ............................................................................................7.5 g - Xylose ............................................................................................3.75 g - Sodium desoxycholate .....................................................................1.0 g - Sodium chloride ...............................................................................5.0 g - Sodium thiosulfate ...........................................................................6.8 g - Ferric ammonium citrate ..................................................................0.8 g - Phenol red...................................................................................80.0 mg - Bacteriological agar .......................................................................12.5 g pH of the ready-to-use medium at 25C : 7.4 0.2.

QUALITY CONTROL - Dehydrated medium : pinkish powder, free-flowing and homogeneous. - Prepared medium : orange red agar. - Typical culture response after 48 hours of incubation at 37C : Microorganisms Salmonella Typhimurium Salmonella Enteritidis Escherichia coli Enterococcus faecalis Staphylococcus aureus ATCC 14028 CIP 82.97 ATCC 25922 ATCC 29212 ATCC 25923 Growth good, score 2 good, score 2 weak, score 0-1 inhibited, score 0 inhibited, score 0 Characteristics red colonies with black center red colonies with black center yellow colonies

STORAGE / SHELF LIFE Dehydrated medium : 2-30C. - The expiration date is indicated on the label. Prepared medium (benchmark value*) : - Media in plates : 8 days at 2-8C. Pre-poured media in Petri plates : - Store between 2-8C, shielded from light. - The expiration date is indicated on the label.

PACKAGING Pre-poured media in Petri plates ( 90 mm) : - 20 plates Dehydrated medium : - 500 g bottle

Code

BM08708 BK168HA

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PHOTO SUPPORT: Product reference: BK168HA, BM08708 Media used for: Isolation of pathogenic Salmonella in food and water samples, and isolation of Shigella according to corresponding standards.

Salmonella Characteristic colony Red colour with a black precipitate in the center

Xylose Lysine Desoxycholate (XLD) Agar

Ref : BM08708 Incubation 24 hours at 37C (surface) Characteristics: Red colonies with a black precipitate in the center (lysine decarboxylation and production of hydrogen sulfide)

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BIBLIOGRAPHY
TAYLOR, W.I. 1965. Isolation of Shigellae. I. Xylose lysine agars; new media for isolation of enteric pathogens. American Journal of Clin. Path., 44(4) : 471-475. XP CEN ISO/TS 11133-2 (V 08-104-2). Janvier 2004. Microbiologie des aliments. Guide pour la prparation et la production des milieux de culture. Partie 2 : Guide gnral pour les essais de performance des milieux de culture. NF EN ISO 21567 (V 08-411). Mars 2005. Microbiologie des aliments. Mthode horizontale pour la recherche de Shigella spp. FD/CEN/TR 15215-2 (X 33-038-2). Avril 2006. Caractrisation des boues. Dtection et dnombrement de Salmonella spp. dans les boues, les sols, les amendements du sol, les supports de culture et biodchets. Partie 2 : Mthode par enrichissement en milieu liquide slnite-cystine puis en milieu de Rapport-Vassiliadis pour la dtermination semiquantitative par la mthode du Nombre le Plus Probable (NPP). FD/CEN/TR 15215-3 (X 33-038-3). Avril 2006. Caractrisation des boues. Dtection et dnombrement de Salmonella spp. dans les boues, les sols, les amendements du sol, les supports de culture et les biodchets. Partie 3 : Prsence/absence par enrichissement en milieu liquide peptone-novobiocine puis sur milieu Rapport-Vassiliadis. NF U 47-100. Juillet 2007. Mthodes danalyse en sant animale. Recherche par lisolement et identification de tout srovar ou de srovar(s) spcifi(s) de salmonelles dans lenvironnement des productions animales. NF EN ISO 6579 (V 08-013). Mars 2007. Microbiologie des aliments. Mthode horizontale pour la recherche des Salmonella spp. NF U 47-101. Novembre 2007. Mthodes danalyse en sant animale. Isolement et identification de tout srovar ou de srovar(s) spcifi(s) de salmonelles chez les oiseaux. NF U 47-102. Janvier 2008. Mthodes danalyse en sant animale. Isolement et identification de tout srovar ou de srovar(s) spcifi(s) de salmonelles chez les mammifres. NF EN ISO 6579/A1 (V 08-013/A1). Janvier 2008. Microbiologie des aliments. Mthode horizontale pour la recherche des Salmonella spp.. Amendement 1 Annexe D : recherche de Salmonella spp. dans les matires fcales des animaux et dans des chantillons au stade de la production primaire. NF ISO 19250 (T 90-435). Octobre 2010. Qualit de leau. Recherche de Salmonella spp.

*Benchmark value refers to the expected shelf life when prepared under standard laboratory conditions following manufacturers 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 2010-10-12. They are susceptible to modification at any time, without warning. Code document : BK168/A/2003-07 : 7.

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