why is citrate test done on agar slants + course hero

by Prof. Stanford Champlin 3 min read

What is the purpose of citrate agar?

Citrate Test. Simmons citrate agar tests the ability of organisms to utilize citrate as a carbon source. Simmons citrate agar contains sodium citrate as the sole source of carbon, ammonium dihydrogen phosphate as the sole source of nitrogen, other nutrients, and the pH indicator bromthymol blue.

What is the principle of Simmons citrate agar?

Sep 17, 2015 · Citrate agar is used to test an organism’s ability to utilize citrate as a source of energy. The medium contains citrate as the sole carbon source and inorganic ammonium salts (NH4H2PO4) as the sole source of nitrogen. Bacteria that can grow on this medium produce an enzyme, citrate-permease, capable of converting citrate to pyruvate.

What is the purpose of the Simmons citrate test?

Jan 10, 2022 · Citrate agar is used to test the ability of an organism to utilize citrate as a source of energy. The agar medium contains citrate as the sole carbon source and inorganic ammonium salts as the sole source of nitrogen.

Do citrate negative organisms change color on Slant agar?

Dec 13, 2019 · Procedure of citrate utilization test: Inoculate Simmons citrate agar lightly on the slant by touching the tip of a needle to a colony that is 18 to 24 hours old. Incubate at 35°C to 37°C for 18 to 24 hours. Some organisms may require up to 7 days of incubation due to their limited rate of growth on citrate medium.

What does the citrate test test for?

Citrate testing is used to determine the ability of the bacteria to use sodium citrate as the only source of carbon and inorganic ammonium hydrogen phosphate (NH4H2PO4) as a source of nitrogen.

Is the citrate agar slant selective or differential?

Simmons citrate agar is a selective and differential medium that tests for an organism's ability to use citrate as a sole carbon source and ammonium ions as the sole nitrogen source.Jan 5, 2022

What does a citrate test tell you about a bacterial sample?

Citrate agar is used to test an organism's ability to utilize citrate as a source of energy. The medium contains citrate as the sole carbon source and inorganic ammonium salts (NH4H2PO4) as the sole source of nitrogen.Jun 14, 2019

Why does a positive Simmons citrate agar result in the slant turning blue?

Growth is indicative of utilization of citrate, an intermediate metabolite in the Krebs cycle. When the bacteria metabolize citrate, the ammonium salts are broken down to ammonia, which increases alkalinity. The shift in pH turns the bromthymol blue indicator in the medium from green to blue above pH 7.6.Aug 15, 2019

Why is the citrate test selective?

It is useful for selecting for organisms that use citrate as its main carbon and energy source. It is a defined, selective and differential medium that tests for an organism's ability to use citrate as a sole carbon source and ammonium ions as the sole nitrogen source.

Why is citrate agar selective?

The citrate utilization test is selective because only certain bacteria can utilize citrate in place of a fermentable carbohydrate.

What is the purpose of citrate utilization test what are the products of this reaction and how are these products detected?

Citrate utilization test is used to determine the ability of bacteria to utilize sodium citrate as its only carbon source and inorganic ammonium dihydrogen phosphate (NH4H2PO4) as the sole nitrogen source.

What bacteria test positive for citrate?

Klebsiella pneumoniae and Proteus mirabilis are examples of citrate positive organisms. Escherichia coli and Shigella dysenteriae are citrate negative.

Why is citrate called a defined medium?

it shows that the organism can produce citrate enzyme. The simmons citrate agar is used to differentiate between gram negative bacteria on the basis of citrate utilization and if that bacteria can use citrate. Defined medium means you know everything in the medium.

Why might an organism with citrate lyase not appear positive on a citrate slant?

Citrate lyase, which catalyzes the conversion of citrate into oxaloacetate and acetate, is associated with how citrate is used, not how it gets into the cell. So, an organism could be citrate-negative (it doesn't make citrate permease), but still be citrate lyase-positive.

Is Simmons citrate agar defined?

Simmons Citrate Agar is a defined medium that tests for an organism's ability to use citrate as its carbon source and ammonia as its nitrogen source. Only bacteria that can transport citrate into the cell from the medium grow well in Citrate Agar.Mar 19, 2021

Principle of Citrate Utilization Test

Citrate agar is used to test an organism’s ability to utilize citrate as a source of energy. The medium contains citrate as the sole carbon source...

Media Used in Citrate Utilization Test

CompositionDeionized water = 1,000 mlFinal pH 6.9 +/- 0.2 at 25 degrees C.Preparation 1. Dissolve above salts in deionized water. 2. Adjust pH to 6...

Procedure of Citrate Utilization Test

1. Streak the slant back and forth with a light inoculum picked from the center of a well-isolated colony. 2. Incubate aerobically at 35 to 37C fo...

Quality Control of Citrate Utilization Test

Citrate Positive: Klebsiella pneumoniae ATCC 13883 (growth; blue color)Citrate Negative: Escherichia coli ATCC 25922 (no growth or trace of growth)

What is citrate agar used for?

Citrate agar is used to test an organism’s ability to utilize citrate as a source of energy. The medium contains citrate as the sole carbon source and inorganic ammonium salts (NH4H2PO4) as the sole source of nitrogen.

Can organisms grow on citrate?

Some organisms are capable of growth on citrate and do not produce a color change. Growth is considered a positive citrate utilization test, even in the absence of a color change.

What is Citrate Utilization Test?

The citrate utilization test is a part of the IMViC test ( Indole, Methyl Red, Vogues-Proskauer, and Citrate Test) that differentiates organisms on the basis of their ability to use citrate as a sole source of energy.

Principle of Citrate Utilization Test

Citrate agar is used to test the ability of an organism to utilize citrate as a source of energy.

Media, Reagents, and Supplies Used

Simmon’s Citrate agar is used as the medium to test the ability of an organism to utilize citrate as a sole source of energy.

Procedure of Citrate Utilization Test

In a beaker, 24.28 grams of the dehydrated powder or lab-prepared media is added to 1000 milliliters of pure distilled or deionized water.

Result Interpretation of Citrate Utilization Test

Figure: Citrate Utilization Test. Image Source: Bailey and Scott’s Diagnostic Microbiology. Elsevier.

Uses of Citrate Utilization Test

Citrate test is used to determine the ability of an organism to utilize citrate as a sole source of energy.

Limitations of Citrate Utilization Test

Luxuriant growth on the slant without an accompanying color change may indicate a positive test. However, if the agar does not turn blue on further incubation, the test should be repeated with a smaller inoculum.

What is Simmons Citrate Agar?

Simmons Citrate Agar is an agar medium used for the differentiation of Enterobacteriaceae based on the utilization of citrate as the sole source of carbon. In the early 1920s, Koser developed a liquid medium formulation for the differentiation of fecal coliforms from the coliform group. Simmons later modified this formulation to produce ...

What is Simmons agar used for?

Simmons Citrate Agar may be used to differentiate citrate-positive Salmonella enteritidis and members of Salmonella subgenus II, III and IV from the citrate-negative Salmonella typhi, Salmonella paratyphi A, Salmonella pullorum and Salmonella gallinarum. Simmons Citrate Agar is primarily used to aid in the identification of Enterobacteriaceae.

Can organisms grow on citrate?

Some organisms are capable of growth on citrate and do not produce a color change. Growth is considered a positive citrate utilization test, even in the absence of a color change. Tests with equivocal results should be repeated. The reactions of this medium alone are not sufficient for identification to the species level.

Can bacteria grow on citrate?

Organisms capable of utilizing ammonium dihydrogen phosphate and citrate will grow unrestricted on this medium. If citrate can be used, the microbe will accumulate alkaline/basic byproducts. Bacteria that can grow on this medium produce an enzyme, citrate-permease, capable of converting citrate to pyruvate.

What is a slant tube?

A slant tube is a test tube prepared by pouring in a melted agar medium while the tube is tilted. This creates a slanted surface on which to inoculate bacteria. Creating a large plug of agar medium and capping the tube prevents desiccation and allows for the long-term storage of living bacterial cultures. It is essential that the tubes and medium ...

What do bacteria need to survive?

Bacteria require several different substances to live including water, a source of energy, carbon, sulfur, nitrogen, phosphorus, vitamins and minerals. They can grow in liquid or solid mediums, but the requirements of some species are quite specific.

What is Simmons citrate agar?

Simmons citrate agar tests the ability of organisms to utilize citrate as a carbon source. Simmons citrate agar contains sodium citrate as the sole source of carbon, ammonium dihydrogen phosphate as the sole source of nitrogen, other nutrients, and the pH indicator bromthymol blue. This test is part of the IMViC tests and is helpful in differentiating the Enterobacteriaceae .

What pH is bromthymol blue?

At pH 7.5 or above , bromthymol blue turns royal blue. At a neutral pH, bromthymol blue is green, as evidenced by the uninoculated media.

How to sterilize wire loops?

Wire loops and needles are made from inert metals such as platinum. They are extremely durable and can be sterilized by incineration. To do this, place the loop or needle into the flame of a Bunsen burner (the hottest portion is in the blue flame). Allow the loop or needle to get red hot, and then remove. Make sure the transfer instrument is cool before inserting into a culture for inoculation. You will do this prior to any inoculation and after any inoculation.

What is an aseptic technique?

Aseptic technique refers to protocols and procedures you use to insure that no microbes contaminate your experiment and that the microbes you are working with do not contaminate you, your colleagues, or working environment, or your loved ones at home! Because microorganisms are ubiquitous, you need to assume that every environment is replete with them. This laboratory exercise will introduce you to the procedures required to ensure a safe and aseptic laboratory. Before beginning today’s laboratory exercise, become acquainted with the terminology and techniques:

Do you need to remove the cap of a transfer tube before inserting a cooled transfer instrument?

Prior to inserting a cooled transfer instrument into a culture tube, you need to remove the cap and flame the lip of the tube. This will help prevent contamination with transient and airborne microbes. You will do this before and after any contact with a transfer instrument.

What is Simmons citrate agar used for?

Simmons’ citrate agar is a selective and differential medium that tests for an organism’s ability to use citrate as a sole carbon source and ammonium ions as the sole nitrogen source. It is used for differentiating gram-negative bacteria on the basis of citrate utilization. It is useful for selecting organisms that use citrate as its main carbon ...

What is citrate used for?

It is useful for selecting organisms that use citrate as its main carbon and energy source. Initially the citrate medium was developed by Koser containing ammonium salt as the only nitrogen source and citrate as the only carbon source for differentiating Escherichia coli and Enterobacter aerogenes by IMViC tests.

What is the sole nitrogen source?

Ammonium dihydrogen phosphate and sodium citrate serve as the sole nitrogen and carbon source respectively. Dipotassium Phosphate acts as a buffer. Sodium Chloride maintains the osmotic balance of the medium. Magnesium Sulfate is a cofactor for a variety of metabolic reactions. Bromothymol blue is the pH indicator.

What is the enzyme that produces pyruvate?

Bacteria that can grow on this medium produce an enzyme, citrate-permease, capable of converting citrate to pyruvate. Pyruvate can then enter the organism’s metabolic cycle for the production of energy. Growth is indicative of utilization of citrate, an intermediate metabolite in the Krebs cycle.

Is magnesium sulfate a cofactor?

Magnesium Sulfate is a cofactor for a variety of metabolic reactions. Bromothymol blue is the pH indicator. Bacteriological agar is the solidifying agent. Organisms capable of utilizing ammonium dihydrogen phosphate and citrate will grow unrestricted on this medium.