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Although it could be used in other formats (e.g., Petri plates), Simmons' citrate agar is often used in slants or slopes in test tubes. One advantage of using slants over Petri dishes is access to oxygen, which is required by citrate metabolism. Simmons’ agar can be bought from suppliers as ready-made powders or slants.
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. This results in a ...
These tubes are commonly sealed with a rubber stopper and often have a specific additive placed in the tube with the stopper color indicating the additive. For example, a blue-top tube is a 5 ml test tube containing sodium citrate as an anticoagulant, used to collect blood for coagulation and glucose-6-phosphate dehydrogenase testing. [5]
This test uses Simmon's citrate agar to determine the ability of a microorganism to use citrate as its sole carbon and energy source. 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.
Stab cultures are similar to agar plates, but are formed by solid agar in a test tube. Bacteria is introduced via an inoculation needle or a pipette tip being stabbed into the center of the agar. Bacteria grow in the punctured area. [ 11 ]
The plates are incubated for 12 hours up to several days, depending on the test that is performed. Commonly used types of agar plates include: Red blood cells on an agar plate are used to diagnose infection. On the left is a positive Staphylococcus infection, on the right a positive Streptococcus culture.
Stab Cultures after inoculation by inoculation needle with E. coli from left to right: TSI, Soft nutrient, Simmons Citrate agar. Inoculation of the microorganisms will be done directly after the transfer from culture to inoculation needle. The inoculum is commonly inoculated to broth cultures, slant cultures, plate cultures, and stab cultures.
This type of test is considered a diffusion test. [18] In agar and broth dilution methods, bacteria are placed in multiple small tubes with different concentrations of antibiotics. [14] Whether a bacterium is sensitive or not is determined by visual inspection or automatic optical methods, after a period of incubation. [5]