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Simmons' citrate agar is used for differentiating gram-negative bacteria on the basis of citrate utilization, [1] especially for distinguishing Gammaproteobacteria of the family Enterobacteriaceae or even between species of the same genus. [2]
Use of citrate involves the enzyme citrate lyase, which breaks down citrate to oxaloacetate and acetate. Oxaloacetate is further broken down to pyruvate and carbon dioxide (CO 2 ). Production of sodium bicarbonate (NaHCO 3 ) as well as ammonia (NH 3 ) from the use of sodium citrate and ammonium salts results in alkaline pH.
The species C. amalonaticus, C. koseri, and C. freundii can use citrate as a sole carbon source. Citrobacter species are differentiated by their ability to convert tryptophan to indole (C. koseri is the only citrobacter to be commonly indole-positive), ferment lactose (C. koseri is a lactose fermentor), and use malonate.
The agar contains citrate and ammonium ions (nitrogen source) and bromothymol blue (BTB) as a pH indicator. [2] Bromothymol blue was added in order to reduce false positives. The citrate agar is green before inoculation, and turns blue, because of BTB as a positive test indicator, meaning citrate is utilized.
Klebsiella aerogenes, [2] previously known as Enterobacter aerogenes, is a Gram-negative, oxidase-negative, catalase-positive, citrate-positive, indole-negative, rod-shaped bacterium. [3] Capable of motility via peritrichous flagella, [ 4 ] it is approximately one to three microns in length.
Positive for the phenylalanine test and the Harnstoff urea test P. vulgaris can test positive or negative for citrate. All combine for a Biocode ID of 31406, (Biocode ID 31402, 31404, 31407 all resulting in P. vulgaris with asymptomatic results) for use in the Interpretation Guide/Computer Coding and Identification System.
TSI agar slant results: (from left) preinoculated (as control), P. aeruginosa, E. coli, Salmonella Typhimurium, Shigella flexneri The Triple Sugar Iron (TSI) test is a microbiological test roughly named for its ability to test a microorganism's ability to ferment sugars and to produce hydrogen sulfide. [1]
Enterobacteriaceae is a large family of Gram-negative bacteria.It includes over 30 genera and more than 100 species. Its classification above the level of family is still a subject of debate, but one classification places it in the order Enterobacterales of the class Gammaproteobacteria in the phylum Pseudomonadota.