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This medium can be used as a screening method for the differentiation of enterobacteriaceae based on the ability of some species to utilise citrate as a sole source of carbon. It is often used as a screening test for Klebsiella pneumoniae (positive reaction) while Escherichia coli is negative. Species that metabolize citrate as their sole source of carbon and ammonium as the sole source of nitrogen cause an increase in alkalinity of the medium resulting in a colour change from green to blue due to the presence of the pH indicator bromthymole blue. -
This medium was originally described by Slanetz and Bartley(1) for the enumeration of enterococci from water samples using membrane filtration. The medium has also become useful as a direct plating medium. Tryptose and yeast extract provide the required carbon, nitrogen and vitamins. Glucose is a fermentable carbohydrate. The di-potassium hydrogen phosphate acts as a buffer. Selectivity is achieved through the addition of sodium azide which is used to suppress the growth of Gram-negative organisms. The medium contains tetrazolium chloride, which is reduced by enterococci to the insoluble red dye formazan resulting in dark red colonies of enterococci on the agar. It should be noted that this reaction is not exclusive to enterococci and colonies should be confirmed by additional testing e.g. aesculin hydrolysis.
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Sorbitol MacConkey agar is a differential medium for the isolation of Escherichia coli 0157:H7 based on the formulation by Rappaport and Henig. It differs from other MacConkey mediums in that lactose has been replaced by sorbitol. As Escherichia coli 0157:H7 does not ferment sorbitol it produces pale translucent colonies whereas most other strains of Escherichia coli are sorbitol positive and produce pink colonies. Although it should be noted that colonies that are sorbitol positive can revert and possibly be mistaken as sorbitol negative. Tryptone and meat peptone provide the required carbon, nitrogen and vitamins. Sorbitol is a fermentable carbohydrate and neutral red is a pH indicator. Bile Salts no.3 and crystal violet are selective agents and together inhibit Gram-positive cocci. Sodium chloride maintains the osmotic balance. If required, the selectivity of the medium may be increased by the addition of cefixime (0.05mg/L) and potassium tellurite (2.5mg/L). Related Supplements : LS0013 Escherichia coli 0157 Selective Supplement
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Soy peptone is manufactured from the enzymatic hydrolysis of soybean. This product provides a good source of nitrogen, carbohydrates, and vitamins. It is recommended for use in microbiological media for the detection and isolation of a wide variety of bacteria and fungi.
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Trichomonas medium is based on the formula described by Kupferburg, Johnson and Sprince for the selective isolation of Trichomonas spp. (1) The tryptone and liver extract act as carbon, nitrogen and vitamin sources in this medium. Maltose is a fermentable carbohydrate. The agar and cysteine HCl reduce the oxygen tension in the medium which aids the growth of trichmonads. Methylene blue is a redox indictor and allows for the visualisation of any significant oxygen diffusion in the medium. The addition of selective agents such as chloramphenicol is recommended to inhibit bacterial species that may be present in specimens. References 1) Kupferberg, A.B. Johnson, G., and Sprince, H. 1948. Proc. Soc. Exper. Biol. Med., 67:304-308
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Triple sugar iron agar is used to differentiate between some of the enterobacteriacae on the basis of four reactions: fermentation of lactose, glucose and sucrose and the production of hydrogen sulphide. Beef extract, yeast extract and peptone provide the required nitrogen, carbon and vitamins. Lactose, sucrose and glucose are carbohydrates. Acid production from their fermentation is detected by the phenol red pH indictor. Sodium thiosulphate is reduced to hydrogen sulphide which is detected by the ferric citrate indicator. Sodium chloride maintains osmotic balance.
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Tryptone is obtained by pancreatic digestion of casein. Casein is the main protein of milk and is a rich source of amino acid and nitrogen. This product can be used in preparing microbiological culture media providing nitrogen, vitamins, minerals and amino acids. Due to the high tryptophan content in tryptone it can be used in detecting indole production.
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Recent developments in culture media have given rise to the use of chromogenic substrates as a means of differentiating bacteria particularly among the coliform group of organisms. This is one such medium and has been developed as a selective medium for the isolation and enumeration of Escherichia coli without the need for membranes or pre-incubation. Based on the formulation of Tryptone Bile Agar it incorporates a chromogenic substrate, X-Glucuronide, to detect the ß-glucuronidase enzyme which is specific for the majority of E. coli strains. Approximately, 3-4% of E. coli are glucuronidase negative including E. coli O157.(1) The advantage of the chromogenic substrate is that the reaction is concentrated within the colony resulting in distinctive blue/green colonies of E. coli while the other coliforms produce cream colonies. The tryptone provides the required carbon, nitrogen and vitamins. Bile salts No.3 is a selective agent against Gram-positive bacteria. X-glucuronide is a chromogenic substrate. Agar is solidifying agent.
