How To Use XLD Agar For Isolation

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PRODUCT DATA SHEET www.titanmedia.in Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Corp office Address: 903-9099 th Floor Bigjos Tower Netaji Subhash Place Delhi-110034. Page 1 INTENDED USE XLD Agar For selective isolation and enumeration of Salmonella typhi and other Salmonella species. COMPOSITION Ingredients Gm\Ltr. Agar 15.000 Sucrose 7.500 Lactose 7.500 Sodium thiosulphate 6.800 L-Lysine 5.000 Sodium chloride 5.000 Xylose 3.500 Yeast extract 3.000 Sodium deoxycholate 2.500 Ferric ammonium citrate 0.800 Phenol red 0.080 PRODUCT SUMMARY AND EXPLANATION XLD Agar was formulated by Taylor for the isolation and differentiation of enteric pathogens including Salmonella typhi from other Salmonella species. XLD Agar has been recommended for the identification of Enterobacteriaceae and for the microbiological testing of foods water and dairy products. The media formulation does not allow the over growth of other organisms over Salmonella and Shigella. XLD Agar is one of the media used in the Microbial Limit Tests in the USP EP PRINCIPLE The medium contains Yeast extracts as source of vitamins and minerals. Addition of Sodium deoxycholate acts as a selective agent which is inhibitory to Gram-positive bacteria. It suppresses the growth of other enteric pathogens and enhances the growth of only few enteric bacilli. The medium contains Xylose as a fermentable carbohydrate which is utilized by Salmonella species. The medium pH is changed due to fermentation of Xylose which is detected by indicator Phenol red thus the colony colour turns red. The medium also contains Lactose and Sucrose as the source of fermentable sugar. L-lysine is an essential amino acid source. Lysine is added to differentiate Salmonella Sp. XLD AGAR TM 492

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PRODUCT DATA SHEET www.titanmedia.in Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Corp office Address: 903-9099 th Floor Bigjos Tower Netaji Subhash Place Delhi-110034. Page 2 Sodium chloride helps maintaining the osmotic balance of the cells. This medium also allows differentiation of bacilli based on their ability to produce H 2S it contains Sodium thiosulphate and Ferric ammonium citrate that helps visualizing the black centered colonies on production of hydrogen sulphide in the medium. Agar is added as the solidifying agent. INSTRUCTION FOR USE 1. Dissolve 56.68g in 1000ml distilled water. 2. Gently heat to boil with gentle swirling and dissolve the medium completely. DO NOT AUTOCLAVE. DO NOT RE-HEAT. 3. Cool to 45 - 50˚C prior to dispense into sterile petri plates QUALITY CONTROL SPECIFICATIONS Appearance of Powder: Light yellow to pink colour homogeneous mixture free flowing powder Appearance of prepared medium: Red colour clear to slightly opalescent gel pH at 25°C: 7.4 ± 0.2 INTERPRETATION Cultural response was observed after inoculating 10 3 CFU/ml after incubation at 35-37°C for 18-24 hours. Microorganisms ATCC Inoculum CFU/ml Growth Recovery Appearance of colony Proteus vulgaris 13315 10 3 Good ≥ 50 Grey with black centres Salmonella typhimurium 14028 10 3 Good ≥ 50 Red with black centres Salmonella paratyphi B 8759 10 3 Good ≥ 50 Red with black centres Salmonella enteritidis 13076 10 3 Good ≥ 50 Red with black centres Salmonella typhi 6539 10 3 Good ≥ 50 Red with black centres Shigella dysenteriae 13313 10 3 Good ≥ 50 Red Salmonella paratyphi A 9150 10 3 Good ≥ 50 Red Shigella sonnei 25931 10 3 Fair to Good 30 -40 Red Staphylococcus aureus 6538 10 3 Inhibited 0 - Enterococcus faecalis 29212 10 3 Inhibited 0 -

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PRODUCT DATA SHEET www.titanmedia.in Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Corp office Address: 903-9099 th Floor Bigjos Tower Netaji Subhash Place Delhi-110034. Page 3 Enterobacter aerogenes 13048 10 3 Fair 20 -30 Yellow Enterobacter cloacae 13047 10 3 Fair 20 -30 Yellow STORAGE STABILITY Dehydrated powder hygroscopic in nature store in a dry place in tightly-sealed containers below 25°C and protect from direct Sunlight. Under optimal conditions the medium has a shelf life of 4 years. When the container is opened for the first time note the time and date on the label space provided on the container. After the desired amount of medium has been taken out replace the cap tightly to protect from hydration. REFERENCES 1. Taylor W.I. 1965. Isolation of shigellae. I. Xylose lysine agars new media for isolation of enteric pathogens. Am. J. Clin. Pathol. 44:471-475. 2. Taylor W.I. and B. Harris. 1965. Isolation of shigellae. II. Comparison of plating media and enrichment broths. Am. J. Clin. Pathol. 44:476-479. 3. Taylor W.I. and B. Harris. 1967. Isolation of shigellae III. Comparison of new and traditional media with stool specimens. Am. J. Clin. Pathol. 48:350-355. 4. Taylor W.I. and D. Schelhart. 1967. Isolation of shigellae. IV. Comparison of plating media with stools. Am. J. Clin. Pathol. 48:356-362. 5. Taylor W.I. and D. Schelhart. 1968. Isolation of shigellae. VI. Performance of media with stool specimens. Appl. Microbiol. 16:1387-1393 2. Chadwick P. Delisle G. H and Byer M. 1974 Can. J. Microbiol. 20 1653-1664. 3. Downes F. P. and Ito K. Ed. 2001 Compendium of Methods for the Microbiological examination of Foods 4th Ed. APHA Inc. Washington D.C. 4. Eaton A. D. Clesceri L. S. Rice E. W. and Greenberg A. W. Eds. 2005 Standard Methods for the Examination of Water and Wastewater 21st Ed. APHA Washington D.C. 5. Wehr H. M. and Frank J. H. 2004 Standard Methods for the Microbiological Examination of Dairy Products 17th Ed. APHA Inc. Washington D.C. good-luxuriant good-luxuriant 50 -100 50 -100 50 -100 10³ 10³ 10³ 50 -100 50 -100 50 -100 50 -100 50 -100 6. Williams H. Ed. 2005 Official Methods of Analysis of the Association of Official Analytical Chemists 19th Ed. AOAC Washington D.C. 7. FDA Bacteriological Analytical Manual 2005 18th Ed. AOAC Washington D.C.

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PRODUCT DATA SHEET www.titanmedia.in Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Corp office Address: 903-9099 th Floor Bigjos Tower Netaji Subhash Place Delhi-110034. Page 4 8. Pollock H.M. and B.J. Dahlgren. 1974. Clinical evaluation of enteric media in the primary isolation of Salmonella and Shigella. Appl. Microbiol. 27:197-201. 9 United States Pharmacopeial Convention Inc. The United States pharmacopeia 25/ The national formulary 20 – 2002. United States Pharmacopeial Convention Inc. Rockville Md. 2001. NOTE: Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

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