Xylose Lysine Deoxycholate Agar

$190.00

100 g, 500g, 2 kg
Number of sizes: 3
Type: Powder

Dehydrated Powder

Sub-products

Product Name Description SKU Price Action
Xylose Lysine Deoxycholate Agar 5 plates/pack, 10 plates/pack Number of sizes: 2 Type: Agar Gel

Prepared Media - Plates

XLDA-P $18.00 View Options
Xylose Lysine Deoxycholate Agar 250 mL, 500 mL Number of sizes: 2 Type: Agar Gel

Prepared Media - Ready to Dissolve

XLDA-ML $25.00 View Options
Xylose Lysine Deoxycholate Agar 100 g, 500g, 2 kg Number of sizes: 3 Type: Powder

Dehydrated Powder

XLDA-G $22.00 View Options

Description

Intended Use and Principle of Procedure
Xylose Lysine Deoxycholate Agar conforms with specifications of The United States Pharmacopeia (USP). XL (Xylose Lysine) Agar Base is used for the isolation and differentiation of enteric pathogens and, when supplemented with appropriate additives, as a base for selective enteric media. Xylose Lysine Deoxycholate Agar is the complete Xylose Lysine Desoxycholate Agar, a moderately selective medium recommended for isolation and differentiation of enteric pathogens, especially Shigella species.

Xylose is incorporated into the medium since it is fermented by practically all enterics except for the shigellae, and this property enables the differentiation of Shigella species. Lysine is included to enable the Salmonella group to be differentiated from the nonpathogens since, without lysine, salmonellae rapidly would ferment the xylose and be indistinguishable from nonpathogenic species. After the salmonellae exhaust the supply of xylose, the lysine is attacked via the enzyme, lysine decarboxylase, with reversion to an alkaline pH which mimics the Shigella reaction. To prevent similar reversion by lysinepositive coliforms, lactose and sucrose (saccharose) were added to produce acid in excess.1 To add to the differentiating ability of the formulation, an H2S indicator system, consisting of sodium thiosulfate and ferric ammonium citrate, is included for the visualization of the hydrogen sulfide produced, resulting in the formation of colonies with black centers. The nonpathogenic H2S producers do not decarboxylate lysine; therefore, the acid reaction produced by them prevents the blackening of the colonies.1 Xylose Lysine Deoxycholate Agar is both a selective and differential medium. It utilizes sodium desoxycholate as the selective agent and, therefore, it is inhibitory to gram-positive microorganisms.

 

Summary and Explanation
A wide variety of media have been developed to aid in the selective isolation and differentiation of enteric pathogens. Due to the large numbers of different microbial species and strains with varying nutritional requirements and chemical resistance patterns, investigators have developed various formulae to meet general as well as specific needs relative to isolation and identification of the microorganisms. XL Agar Base was developed by Taylor1 for the nonselective isolation and differentiation of gram-negative enteric bacilli.

It is particularly recommended for obtaining counts of enteric organisms. This medium can be rendered moderately selective for enteric pathogens, particularly Shigella, by the addition of sodium desoxycholate (2.5 g/L) to make Xylose Lysine Deoxycholate Agar.1 XL Agar Base can be made selective for Salmonella by adding 1.25 mL/L of 1% aqueous brilliant green to the base prior to autoclaving. Its use is recommended for Salmonella isolation after selenite or tetrathionate enrichment in food analysis; both coliforms and Shigella are inhibited.1 XLD Agar was developed by Taylor in order to increase the efficiency of the isolation and identification of enteric pathogens, particularly Shigella. 1 The pathogens are differentiated not only from the nonpathogenic lactose fermenters but also from many nonpathogens which do not ferment lactose or sucrose. Additionally, the medium was formulated to increase the frequency of growth of the more fastidious pathogens,1 which in other formulations have often failed to grow due to the inclusion of excessively toxic inhibitors. The results obtained in a number of clinical evaluations have supported the claim for the relatively high efficiency of XLD Agar in the primary isolation of Shigella and Salmonella. 2-6 Xylose Lysine Deoxycholate Agar is included in the USP microbial limit test for screening specimens for the presence or absence of Salmonella7 and is recommended for the testing of foods, dairy products and water.

 

Formulae
 

Xylose 3.75g
L-Lysine 5.0g
Lactose 7.5g
Saccharose 7.5g
Sodium Chloride 5.0g
Yeast Extract 3.0g
Phenol Red 0.08g
Sodium Desoxycholate 2.5g
Sodium Thiosulfate 6.8g
Ferric Ammonium Citrate 0.8g
Agar 15.0g

 

Procedure
Use standard procedures to obtain isolated colonies from specimens. A nonselective medium should also be streaked to increase the chance of recovery when the population of gramnegative organisms is low and to provide an indication of other organisms present in the specimen. Incubate plates, protected from light, at 35 ± 2°C for 18-24 hours. Colonies on XLD agar may require 48 hours incubation for full color development.

 

Directions for Preparation from Dehydrated Product
1. Suspend the powder in 1 L of purified water: Difco XLD Agar – 57 g; Mix thoroughly.

2. Heat with agitation just until the medium boils. DO NOT OVERHEAT. DO NOT AUTOCLAVE.

3. Cool to 45-50°C in a water bath and use immediately. Overheating causes precipitation.

4. Test samples of the finished product for performance using stable, typical control cultures.

 

Identification Specification and Cultural Response
 

Dehydrated Appearance: Fine, homogeneous, free of extraneous material
 

 

Prepared Appearance:

 

 

Solution:

 

Medium to dark, orange-red to rose-red, clear to slightly hazy

 

5.5% solution, soluble in purified water upon boiling. Solution is medium to dark, orange-red to rose-red, clear to slightly hazy

 

pH:

 

7.4 ± 0.2
Cultural Response

 

Prepare the medium per label directions. Inoculate and incubate at 35 ± 2°C for 18-24 hours
Reaction 5.5%
Solution 25°C

 

 

Microorganism ATCC CFU Recovery
Escherichia coli 25922 102-104 Good
Salmonella choleraesuis subsp. choleraesuis serotype Typhimurium 14028 102-104 Good
Shigella flexneri 12022 102-104 Good

 

Expected Results
Degradation of xylose, lactose and sucrose generates acid products, causing a color change in the medium from red to yellow. Hydrogen sulfide production under alkaline conditions causes colonies to develop black centers. This reaction is inhibited by the acid conditions that accompany carbohydrate fermentation. Lysine decarboxylation in the absence of lactose and sucrose fermentation causes reversion to an alkaline condition and the color of the medium changes back to red. Typical colonial morphology and reactions on XLD Agar are as follows:

 

E.coli …………………………….. Large, flat, yellow; some strains may be inhibited

Enterobacter/ Klebsiella ……. Mucoid, yellow

Proteus………………………….. Red to yellow; most strains have black centers

Salmonella …………………….. Red-yellow with black centers

Shigella, Salmonella H2 S-negative ………………….. Red

Pseudomonas…………………. Red

Gram-positive bacteria …….. No growth to slight growth

 

Limitations of the Procedure
1. Red, false-positive colonies may occur with some Proteus and Pseudomonas species.

2. Incubation in excess of 48 hours may lead to false-positive results.

3. S. paratyphi A, S. choleraesuis, S. pullorum and S. gallinarum may form red colonies without black centers, thus resembling Shigella species.

4. Some Proteus strains will give black-centered colonies on XLD Agar.

 

References
1. Taylor. 1965. Am. J. Clin. Pathol. 44:471.

2. Taylor and Harris. 1965. Am. J. Clin. Pathol. 44:476.

3. Taylor and Harris. 1967. Am. J. Clin. Pathol. 48:350.

4. Taylor and Schelhart. 1967. Am. J. Clin. Pathol. 48:356.

5. Taylor and Schelhart. 1968. Appl. Microbiol. 16:1387.

6. Pollock and Dahlgren. 1974. Appl. Microbiol. 27:197.

7. United States Pharmacopeial Convention, Inc. 2001. The United States pharmacopeia 25/The national formulary 20 – 2002. United States Pharmacopeial Convention, Inc., Rockville, Md.

8. U.S. Food and Drug Adminsitration. 1995. Bacteriological analytical manual, 8th ed. AOAC International, Gaithersburg, Md.

9. Horwitz (ed.). 2000. Official methods of analysis of AOAC International, 17th ed. AOAC International, Gaithersburg, Md.

10. Downes and Ito (ed.). 2001. Compendium of methods for the microbiological examination of foods, 4th ed. American Public Health Association, Washington, D.C.

11. Marshall (ed.). 1993. Standard methods for the examination of dairy products, 16th ed. American Public Health Association, Washington, D.C.

12. Clesceri, Greenberg and Eaton (ed.). 1998. Standard methods for the examination of water and wastewater, 20th ed. American Public Health Association, Washington, D.C.

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