MacConkey agar

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With an active bacterial culture Proteus McConkey.jpg
With an active bacterial culture
Lactose Lactose v2.svg
Lactose
The organism on the left is a lactose fermenter, as evidenced by the pink color. The organism on the right produced no color, so it does not appear to be a lactose fermenter. MacConkey agar with LF and LF colonies.jpg
The organism on the left is a lactose fermenter, as evidenced by the pink color. The organism on the right produced no color, so it does not appear to be a lactose fermenter.

MacConkey agar is a selective and differential culture medium for bacteria. It is designed to selectively isolate Gram-negative and enteric (normally found in the intestinal tract) bacteria and differentiate them based on lactose fermentation. [1] Lactose fermenters turn red or pink on MacConkey agar, and nonfermenters do not change color. The media inhibits growth of Gram-positive organisms with crystal violet and bile salts, allowing for the selection and isolation of gram-negative bacteria. The media detects lactose fermentation by enteric bacteria with the pH indicator neutral red. [2]

Contents

Contents

It contains bile salts (to inhibit most Gram-positive bacteria), crystal violet dye (which also inhibits certain Gram-positive bacteria), and neutral red dye (which turns pink if the microbes are fermenting lactose).

Composition: [3]

There are many variations of MacConkey agar depending on the need. If the spreading or swarming of Proteus species is not required, sodium chloride is omitted. Crystal violet at a concentration of 0.0001% (0.001 g per litre) is included when needing to check if Gram-positive bacteria are inhibited. MacConkey with sorbitol is used to isolate E. coli O157, an enteric pathogen. [4]

History

The medium was developed by Alfred Theodore MacConkey while working as a bacteriologist for the Royal Commission on Sewage Disposal. [5]

Uses

Using neutral red pH indicator, the agar distinguishes those Gram-negative bacteria that can ferment the sugar lactose (Lac+) from those that cannot (Lac-).

This medium is also known as an "indicator medium" and a "low selective medium". Presence of bile salts inhibits swarming by Proteus species.

Lac positive

By utilizing the lactose available in the medium, Lac+ bacteria such as Escherichia coli , Enterobacter and Klebsiella will produce acid, which lowers the pH of the agar below 6.8 and results in the appearance of pink colonies. The bile salts precipitate in the immediate neighborhood of the colony, causing the medium surrounding the colony to become hazy. [6] [7]

Lac negative

Organisms unable to ferment lactose will form normal-colored (i.e., un-dyed) colonies. The medium may also turn yellow. Examples of non-lactose fermenting bacteria include Salmonella , Proteus, and Shigella spp. . [4]

Slow

Some organisms ferment lactose slowly or weakly, and are sometimes put in their own category. These include Serratia [8] and Citrobacter . [9]

Mucoid colonies

Some organisms, especially Klebsiella and Enterobacter, produce mucoid colonies which appear very moist and sticky and slimy. This phenomenon happens because the organism is producing a capsule, which is predominantly made from the lactose sugar in the agar.

Variant

A variant, sorbitol-MacConkey agar, (with the addition of additional selective agents) can assist in the isolation and differentiation of enterohemorrhagic E. coli serotype E. coli O157:H7, by the presence of colorless circular colonies that are non-sorbitol fermenting. [4]

See also

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Sorbitol-MacConkey agar is a variant of traditional MacConkey agar used in the detection of E. coli O157:H7. Traditionally, MacConkey agar has been used to distinguish those bacteria that ferment lactose from those that do not. This is important because gut bacteria, such as Escherichia coli, can typically ferment lactose, while important gut pathogens, such as Salmonella enterica and most shigellas are unable to ferment lactose. Shigella sonnei can ferment lactose, but only after prolonged incubation, so it is referred to as a late-lactose fermenter.

<span class="mw-page-title-main">Cystine–lactose–electrolyte-deficient agar</span>

CLED agar is a valuable non-inhibitory growth medium used in the isolation and differentiation of urinary microbes. It contains cystine and lactose and is electrolyte-deficient; the latter trait prevents the swarming of Proteus species. Cystine promotes the formation of cystine-dependent dwarf colonies. Bromothymol blue is the indicator used in the agar, it changes to yellow in case of acid production during fermentation of lactose or changes to deep blue in case of alkalinization. Lactose-positive bacteria build yellow colonies. Bacteria which decarboxylate L-cystine cause an alkaline reaction and build deep blue colonies.

<span class="mw-page-title-main">XLD agar</span> Selective culture medium

Xylose Lysine Deoxycholate agar is a selective growth medium used in the isolation of Salmonella and Shigella species from clinical samples and from food. The agar was developed by Welton Taylor in 1965. It has a pH of approximately 7.4, leaving it with a bright pink or red appearance due to the indicator phenol red. Sugar fermentation lowers the pH and the phenol red indicator registers this by changing to yellow. Most gut bacteria, including Salmonella, can ferment the sugar xylose to produce acid; Shigella colonies cannot do this and therefore remain red. After exhausting the xylose supply Salmonella colonies will decarboxylate lysine, increasing the pH once again to alkaline and mimicking the red Shigella colonies. Salmonellae metabolise thiosulfate to produce hydrogen sulfide, which leads to the formation of colonies with black centers and allows them to be differentiated from the similarly coloured Shigella colonies.

<span class="mw-page-title-main">Mannitol salt agar</span> Culture medium used in microbiology

Mannitol salt agar or MSA is a commonly used selective and differential growth medium in microbiology. It encourages the growth of a group of certain bacteria while inhibiting the growth of others. It contains a high concentration of salt (NaCl) which is inhibitory to most bacteria - making MSA selective against most Gram-negative and selective for some Gram-positive bacteria that tolerate high salt concentrations. It is also a differential medium for mannitol-fermenting staphylococci, containing the sugar alcohol mannitol and the indicator phenol red, a pH indicator for detecting acid produced by mannitol-fermenting staphylococci. Staphylococcus aureus produces yellow colonies with yellow zones, whereas other coagulase-negative staphylococci produce small pink or red colonies with no colour change to the medium. If an organism can ferment mannitol, an acidic byproduct is formed that causes the phenol red in the agar to turn yellow. It is used for the selective isolation of presumptive pathogenic (pp) Staphylococcus species.

<span class="mw-page-title-main">MRS agar</span> Selective culture medium for Lactobacilli

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<span class="mw-page-title-main">Thiosulfate–citrate–bile salts–sucrose agar</span>

Thiosulfate–citrate–bile salts–sucrose agar, or TCBS agar, is a type of selective agar culture plate that is used in microbiology laboratories to isolate Vibrio species. TCBS agar is highly selective for the isolation of V. cholerae and V. parahaemolyticus as well as other Vibrio species. Apart from TCBS agar, other rapid testing dipsticks like immunochromatographic dipstick is also used in endemic areas such as Asia, Africa and Latin America. Though, TCBS agar study is required for confirmation. This becomes immensely important in cases of gastroenteritis caused by campylobacter species, whose symptoms mimic that of cholera. Since no yellow bacterial growth is observed in case of campylobacter species on TCBS agar, chances of incorrect diagnosis can be rectified. TCBS agar contains high concentrations of sodium thiosulfate and sodium citrate to inhibit the growth of Enterobacteriaceae. Inhibition of gram-positive bacteria is achieved by the incorporation of ox gall, which is a naturally occurring substance containing a mixture of bile salts and sodium cholate, a pure bile salt. Sodium thiosulfate also serves as a sulfur source and its presence, in combination with ferric citrate, allows for the easy detection of hydrogen sulfide production. Saccharose (sucrose) is included as a fermentable carbohydrate for metabolism by Vibrio species. The alkaline pH of the medium enhances the recovery of V. cholerae and inhibits the growth of others. Thymol blue and bromothymol blue are included as indicators of pH changes.

<span class="mw-page-title-main">New York City agar</span>

The NYC medium or GC medium agar is used for isolating Gonococci.

References

  1. "MacConkey Agar". Texas Medical Center. Archived from the original on 2008-11-04.
  2. Anderson, Cindy (2013). Great Adventures in the Microbiology Laboratory (7th ed.). Pearson. pp. 175–176. ISBN   978-1-269-39068-2.
  3. "MacConkey Agar Plates Protocols". Archived from the original on 2010-12-03. Retrieved 2011-03-20.
  4. 1 2 3 Doern, Christopher D. (2018). Pocket Guide to Clinical Microbiology (4th ed.). ASM Press. p. 149. ISBN   9781683670063.
  5. Smith, Kenneth (2019-10-14). "The Origin of MacConkey Agar". American Society for Microbiology . Retrieved 2023-12-01.
  6. MacConkey AT (1905). "Lactose-Fermenting Bacteria in Faeces". J Hyg (Lond). 5 (3): 333–79. doi:10.1017/s002217240000259x. PMC   2236133 . PMID   20474229.
  7. MacConkey AT (1908). "Bile Salt Media and their advantages in some Bacteriological Examinations". J Hyg (Lond). 8 (3): 322–34. doi:10.1017/s0022172400003375. PMC   2167122 . PMID   20474363.
  8. Luis M. De LA Maza; Pezzlo, Marie T.; Janet T. Shigei; Peterson, Ellena M. (2004). Color Atlas of Medical Bacteriology. Washington, D.C: ASM Press. p. 103. ISBN   1-55581-206-6.
  9. "Medmicro Chapter 26". Archived from the original on 2008-07-06. Retrieved 2008-12-11.