Simmons_Citrate_Agar_TD

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Simmons Citrate agar is used to test an organism’s ability to utilize citrate as a source of energy. Ammonium dihydrogen phosphate and sodium citrate serve as the sole nitrogen and carbon source respectively.

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 1 SIMMONS CITRATE AGAR ISO 10273:2003 TM 348 www.tmmedia.in PRODUCT DATA SHEET INTENDED USE Simmons Citrate Agar For differentiation of Enterobacteriaceae on the basis of citrate utilization. COMPOSITION Ingredients Gm\Ltr. Agar 15.000 Sodium chloride 5.000 Sodium citrate 2.000 Dipotassium phosphate 1.000 Ammonium dihydrogen phosphate 1.000 Magnesium sulphate 0.200 Bromothymol blue 0.080 PRODUCT SUMMARY AND EXPLANATION SIMMONS CITRATE AGAR is used for the differentiation between Enterobacteriaceae and the members of aerogenes group on the basis of citrate utilization as sole carbon source. It is recommended for the differentiation of coliforms isolated from water and clinical samples. The medium is virtually a solidified form of Koser citrate medium which in its original form suffered from the disadvantage that false appearance of growth occurred when large inocula were employed. The addition of bromothymol blue indicator to the medium was a distinct improvement. Simmons Citrate Agar complies with the recommendations of the APHA. ISO 10273 recommends this medium for the confirmation of Yersinia enterocolitica. Inoculate and incubate at 30˚C during 24 hours. The medium remains green since Yersinia enterocolitica does not use citrate as the sole source of carbon. Simmons Citrate Agar is recommended for differentiation of enteric Gram-negative bacilli from laboratory specimens water samples and food samples. PRINCIPLE Simmons Citrate agar is used to test an organism’s ability to utilize citrate as a source of energy. Ammonium dihydrogen phosphate and sodium citrate serve as the sole nitrogen and carbon source respectively. Microorganisms also use inorganic ammonium salts as their sole nitrogen source. Metabolism of these salts causes the medium to become alkaline indicated by a change in colour of the pH indicator from green to blue. Bromothymol blue is the pH indicator. Dipotassium Phosphate act as a buffer. Sodium chloride maintains the osmotic balance of the medium. Magnesium Sulfate is a cofactor for a variety of metabolic reactions.

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 2 www.tmmedia.in PRODUCT DATA SHEET INSTRUCTION FOR USE 1. Dissolve 24.28 grams in 1000 ml distilled water. 2. Gently heat with constant stirring to dissolve the medium completely. 3. Mix well and distribute in tubes or flasks. 4. Sterilize by autoclaving at 15 psi pressure 121°C for 15 minutes. 5. After autoclaving allow medium to solidify in a slanted position. QUALITY CONTROL SPECIFICATIONS Appearance of Powder: Cream to yellow colour homogeneous free flowing powder. Appearance of prepared medium: Forest green colour slightly opalescent gel. pH at 25°C: 6.8 ± 0.2. INTERPRETATION: Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours Microorganisms ATCC Inoculum CFU/ml Growth Citrate utilization Enterobacter aerogenes 13048 50 - 100 Good-Luxuriant Positive reaction Blue colour Salmonella enteritidis 13076 50 - 100 Good-Luxuriant Positive reaction Blue colour Salmonella typhimurium 14028 50 - 100 Good- Luxuriant Positive reaction Blue colour Salmonella typhi 6539 50 - 100 Fair - Good Negative reaction Green colour Shigella dysenteriae 13313 ≥ 10 3 Inhibited ------ Escherichia coli 25922 ≥ 10 3 Inhibited ------ Yersinia enterocolitica 27729 ≥ 10 3 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.

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 3 www.tmmedia.in PRODUCT DATA SHEET REFERENCES 1. Koser S. A. 1923. Utilization of the salts of organic acids by the colon-aerogenes group. J. Bacteriol. 8:493. 2. Simmons J. S. 1926. A culture medium for differentiating organisms of typhoid-colon aerogenes groups and for isolation of certain fungi. J. Infect. Dis. 39:209. 3. MacFaddin J. 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria Vol. 1 Williams and Wilkins Baltimore. 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. Revision : 03 / 2017 5. American Public Health Association Standard Methods for the Examination of Dairy Products 1978 14th Ed. Washington D.C. 6. ISO 10273. Microbiology of food and animal feeding stuffs — Horizontal method for the detection of presumptive pathogenic Yersinia enterocolitica NOTE: Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

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