Cetrimide_Agar_Base_TD

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CETRIMIDE AGAR is used for the selective isolation of Pseudomonas aeruginosa from clinical samples. The medium is a modification of the medium proposed by Brown and Lowbury (1965) for the isolation and differentiation of Pseudomonas aeruginosa from various materials.

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Page 1 Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. www.tmmedia.in PRODUCT DATA SHEET INTENDED USE Cetrimide Agar Base For selective isolation of Pseudomonas aeruginosa from clinical samples. COMPOSITION Ingredients Gms/Ltr Pancreatic digest of gelatin 20.000 Agar 15.000 Potassium sulphate 10.000 Magnesium chloride 1.400 Cetrimide 0.300 PRODUCT SUMMARY AND EXPLAINATION CETRIMIDE AGAR is used for the selective isolation of Pseudomonas aeruginosa from clinical samples. The medium is a modification of the medium proposed by Brown and Lowbury 1965 for the isolation and differentiation of Pseudomonas aeruginosa from various materials. The medium contains quaternary ammonium cationic detergent compound like cetrimide which has a property to inhibit all the bacteria except P. aeruginosa thus making the medium selective for Pseudomonas aeruginosa. Cetrimide causes release of nitrogen and phosphorus from bacterial cells other than P. aeruginosa. Cetrimide Agar is recommended in the United States Pharmacopoeia XXVI and European Pharmacopoeia IV for use in Microbial Limit Tests. The formulation is also in the A.O.A.C. guidelines for isolation of Pseudomonas aeruginosa from cosmetics and in the A.O.A.C. method for testing disinfectants on hard surfaces. PRINCIPLE Pancreatic digest of gelatin is a source of carbon and nitrogen for the growth of microorganisms. Agar is a solidifying agent. Sodium chloride maintains osmotic equilibrium in the medium. Magnesium chloride and Potassium sulphate stimulates the bacterium to produce and secrete a pigment called pyocyanin due to which the colony colour turns light green and also they forms a grape like odour. Cetrimide is a quaternary ammonium salt which acts as a cationic detergent that reduces surface tension in the point of contact and has precipitant complexing and denaturing effects on bacterial membrane proteins. It exhibits inhibitory actions on a wide variety of microorganisms including Pseudomonas species other than Pseudomonas aeruginosa. Goto and Enomoto 1970 recommend the addition of 15 µg nalidixic acid/ml to improve the inhibition of the accompanying microbial flora. INSTRUCTIONS FOR USE CETRIMIDE AGAR BASE TM 060

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Page 2 Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. www.tmmedia.in PRODUCT DATA SHEET 1. Dissolve 46.7 grams in 1000 ml distilled water containing 10 ml glycerol. 2. Gently heat to boiling with gentle swirling and dissolve the medium completely. 3. Sterilize by autoclaving at 15 lbs pressure 121°C for 15 minutes. 4. Cool to 45-50°C. 5. If desired rehydrated contents of 1 vial of Nalidixic Selective Supplement TS 020 may be added aseptically to 1000 ml medium. 6. Mix well and pour into sterile Petri plates. CONTROL SPECIFICATIONS Appearance Dehydrated powder: Cream to yellow colour homogenous free flowing powder Appearance of the prepared medium: Light amber colour opalescent gel with slight precipitate. pH at 25°C : 7.2 ± 0.2 INTERPRETATION Cultural characteristics observed after incubation at 35 ± 2°C for 24 hours. Organism ATCC Inoculum CFU/ml Growth Pseudomonas aeruginosa 27853 50-100 Luxuriant Pseudomonas aeruginosa 9027 50-100 Luxuriant Escherichia coli 25922 ≥ 1000 Inhibited Staphylococcus aureus 25923 ≥ 1000 Inhibited 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. Forbes B. A. Sahm A. S. and Weissfeld D. F. Bailey Scotts Diagnostic Microbiology 10th Ed. 1998 Mosby Inc. St. Louis Mo.

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Page 3 Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. www.tmmedia.in PRODUCT DATA SHEET 2. Williams Ed. 2005 Official Methods of Analysis of the Association of Official Analytical Chemists 19th Ed. AOAC Washington D.C. 3. Goto S. a. Enomoto S.: Nalidixic acid cetrimide agar. A new selective plating medium for the selective isolation of Pseudomonas aeruginosa. - Japan J. Microbiol. 14 65-72 1970. 4. Brown V.I. and Lowbury E.J.L. 1965: Use of improved cetrimide agar medium and other culture methods for Pseudomonas aeruginosa. J. Clin. Pathol. 18: 752-756. 5. United States Pharmacopeia 2002. Microbiological Limit Tests United States Pharmacopeia 26th Ed. United States Pharmacopial Convention Rockville M.D. 6. European Pharmacopeia 2002. Microbial Examination of Non-Sterile Products Test for Specified Microorganisms. European Pharmacopoeia 4th Ed. 7. Official Analytical Chemists. 1995. Bacteriological Analytical Manual 8th Ed. AOAC International Gaithersburg M.D. NOTE: Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

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