Hektoen enteric agar

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Hektoen enteric agar
Gram negative Shigella sonnei bacteria which spent 48 hours cultured on Hektoen enteric agar.jpg
Gram negative Shigella sonnei bacteria which spent 48 hours cultured on Hektoen enteric agar (HEK).
AcronymHEK, HE, or HEA
InventorHektoen Institute
ManufacturerHektoen Institute

Hektoen enteric agar (HEK, HE or HEA) is a selective and differential agar [1] primarily used to recover Salmonella and Shigella from patient specimens. HEA contains indicators of lactose fermentation and hydrogen sulfide production; as well as inhibitors to prevent the growth of Gram-positive bacteria. It is named after the Hektoen Institute in Chicago, where researchers developed the agar.

Contents

Use

The definitive use of HEA is to discriminate between Shigella and Salmonella, although many other species may grow on these plates. However, while the other bacteria may be clinically relevant, the assay does not discriminate among them. Effectively, HEA uses a metabolic assay to divide colonies into "Salmonella and Shigella" and "everything else". Use of these plates assumes that the user is not interested in other enteric pathogens such as Klebsiella or Escherichia .

The plates contain various sugar sources (lactose, sucrose, and salicin), none of which can be used by either Shigella or Salmonella, but the medium also includes peptone which can be used as a carbon source. Since most bacteria can use the sugars in preference to peptone, these "uninteresting" bacteria acidify the medium and turn a pH indicator yellow or red. Peptone metabolism by Shigella and Salmonella alkalises the medium, turning a pH indicator blue.

The presence of thiosulfate or ferric ammonium citrate in the medium produces a black precipitate in the presence of H2S, allowing Shigella – which does not produce H2S, and appears as green colonies – to be distinguished from Salmonella – which does produce hydrogen sulfide and appears as black colonies. [2]

Few sulfur-reducing bacteria exist other than Salmonella, which can be isolated from the intestines. Most of these are inhibited on HEA plates by the inclusion of bile salts, so encountering a black colony that is not Salmonella is unusual, although not unheard of. Those that are may be identified as red or yellow colonies with a black centre, indicating that they are fermenting sugar and probably not Salmonella. However, rare strains of Salmonella are capable of lactose fermentation, [3] which will appear in the same way.

See also

Related Research Articles

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Shigellosis is an infection of the intestines caused by Shigella bacteria. Symptoms generally start one to two days after exposure and include diarrhea, fever, abdominal pain, and feeling the need to pass stools even when the bowels are empty. The diarrhea may be bloody. Symptoms typically last five to seven days and it may take several months before bowel habits return entirely to normal. Complications can include reactive arthritis, sepsis, seizures, and hemolytic uremic syndrome.

<span class="mw-page-title-main">Agar plate</span> Petri dish with agar used to culture microbes

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<span class="mw-page-title-main">Microbiological culture</span> Method of allowing microorganisms to multiply in a controlled medium

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<span class="mw-page-title-main">Bacteriological water analysis</span>

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<span class="mw-page-title-main">Coliform bacteria</span> Group of bacterial species

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<span class="mw-page-title-main">Eosin methylene blue</span> Biological stain

Eosin methylene blue is a selective and differential media used for the identification of Gram-negative bacteria, specifically the Enterobacteriaceae. EMB inhibits the growth of most Gram-positive bacteria. EMB is often used to confirm the presence of coliforms in a sample. It contains two dyes, eosin and methylene blue in the ratio of 6:1. EMB is a differential microbiological media, which inhibits the growth of Gram-positive bacteria and differentiates bacteria that ferment lactose from those that do not. Organisms that ferment lactose appear dark/black or green often with "nucleated colonies"—colonies with dark centers. Organisms that do not ferment lactose will appear pink and often mucoid.

<span class="mw-page-title-main">Growth medium</span> Solid, liquid or gel used to grow microorganisms or cells

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Shigella dysenteriae is a species of the rod-shaped bacterial genus Shigella. Shigella species can cause shigellosis. Shigellae are Gram-negative, non-spore-forming, facultatively anaerobic, nonmotile bacteria. S. dysenteriae has the ability to invade and replicate in various species of epithelial cells and enterocytes.

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">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.

Leuconostoc mesenteroides is a species of lactic acid bacteria associated with fermentation, under conditions of salinity and low temperatures. In some cases of vegetable and food storage, it was associated with pathogenicity. L. mesenteroides is approximately 0.5-0.7 µm in diameter and has a length of 0.7-1.2 µm, producing small grayish colonies that are typically less than 1.0 mm in diameter. It is facultatively anaerobic, Gram-positive, non-motile, non-sporogenous, and spherical. It often forms lenticular coccoid cells in pairs and chains, however, it can occasionally form short rods with rounded ends in long chains, as its shape can differ depending on what media the species is grown on. L. mesenteroides grows best at 30 °C, but can survive in temperatures ranging from 10 °C to 30 °C. Its optimum pH is 5.5, but can still show growth in pH of 4.5-7.0.

<span class="mw-page-title-main">TSI slant</span> Differential medium used in microbiology

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<i>Aeromonas salmonicida</i> Species of bacterium

Aeromonas salmonicida is a pathogenic bacterium that severely impacts salmonid populations and other species. It was first discovered in a Bavarian brown trout hatchery by Emmerich and Weibel in 1894. Aeromonas salmonicida's ability to infect a variety of hosts, multiply, and adapt, make it a prime virulent bacterium. A. salmonicida is an etiological agent for furunculosis, a disease that causes sepsis, haemorrhages, muscle lesions, inflammation of the lower intestine, spleen enlargement, and death in freshwater fish populations. It is found worldwide with the exception of South America. The major route of contamination is poor water quality; however, it can also be associated stress factors such as overcrowding, high temperatures, and trauma. Spawning and smolting fish are prime victims of furunculosis due to their immunocompromised state of being.

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

Endo agar is a microbiological growth medium with a faint pink colour. Originally developed for the isolation of Salmonella typhi, it is now used mostly as a coliform medium. Most gram-negative organisms grow well in this medium, while growth of gram-positive organisms is inhibited. Coliform organisms ferment the lactose in this medium, producing a green metallic sheen, whereas non-lactose-fermenting organisms produce clear, colourless colonies, i.e. Salmonella species.

<span class="mw-page-title-main">Thiosulfate–citrate–bile salts–sucrose agar</span> Culture medium used in microbiology

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.

Oxidative/fermentation glucose test is a biological technique. It was developed in 1953 by Hugh and Leifson to be utilized in microbiology to determine the way a microorganism metabolizes a carbohydrate such as glucose (dextrose). OF-glucose deeps contain glucose as a carbohydrate, peptones, bromothymol blue indicator for Hugh-Leifson's OF medium or phenol red for King's OF medium, and 0.5% agar.

Thermotoga naphthophila is a hyperthermophilic, anaerobic, non-spore-forming, rod-shaped fermentative heterotroph, with type strain RKU-10T.

References

  1. Goo VY, Ching GQ, Gooch JM (September 1973). "Comparison of brilliant green agar and Hektoen enteric agar media in the isolation of salmonellae from food products". Appl Microbiol. 26 (3): 288–92. doi:10.1128/am.26.3.288-292.1973. PMC   379776 . PMID   4584576.
  2. Hudzicki, J. "Hektoen Enteric Agar Protocol." American Society for Microbiology Microbe Library Archived 2013-01-18 at the Wayback Machine retrieved Jan 2013
  3. McDonough, PL; Shin, SJ; Lein, DH (March 2000). "Diagnostic and public health dilemma of lactose-fermenting Salmonella enterica serotype Typhimurium in cattle in the Northeastern United States". Journal of Clinical Microbiology. 38 (3): 1221–6. doi:10.1128/JCM.38.3.1221-1226.2000. PMC   86381 . PMID   10699026.