DCA agar ( deoxycholate citrate agar ) is a solid bacteriological growth medium used for isolation of enteric pathogens. [1]
DCA agar ( deoxycholate citrate agar ) is a solid bacteriological growth medium used for isolation of enteric pathogens. [1]
It is particularly useful for the isolation of organisms that cause bacillary dysentery, salmonella strains that cause food poisoning and Salmonella Paratyphi. It is not so selective for Salmonella Typhi. This growth medium is inhibitory to most gut bacteria, in particular species of the genus Proteus , although these species do survive on DCA agar. It is therefore essential that suspected pathogens must be subcultured on a less inhibitory medium prior to identification. Salmonella spp appear to be yellow or colourless colonies, often with a dark centre. As there are many bacteria that also look like Salmonella on DCA, it is widely recommended that more selective agars are used for the identification of Salmonella, namely xylose lysine deoxycholate (XLD) agar. This growth medium is heat-sensitive and should be poured and cooled as soon as possible after addition of the deoxycholate, otherwise it tends to become very soft and difficult to handle. It has a pH of approximately 7.3, and when poured and cooled, appears light to dark pink in colour.
One liter of deoxycholate citrate agar contains: [1]
| Sodium citrate | 20 grams |
| Agar | 17 grams |
| Lactose | 10 grams |
| Solids from meat infusion | 10 grams |
| Peptic digest of animal tissue | 10 grams |
| Sodium deoxycholate | 5 grams |
| Ferric citrate | 1 gram |
| Neutral red | 20 milligrams |
An agar plate is a Petri dish that contains a growth medium solidified with agar, used to culture microorganisms. Sometimes selective compounds are added to influence growth, such as antibiotics.
Bacteriological water analysis is a method of analysing water to estimate the numbers of bacteria present and, if needed, to find out what sort of bacteria they are. It represents one aspect of water quality. It is a microbiological analytical procedure which uses samples of water and from these samples determines the concentration of bacteria. It is then possible to draw inferences about the suitability of the water for use from these concentrations. This process is used, for example, to routinely confirm that water is safe for human consumption or that bathing and recreational waters are safe to use.
Coliform bacteria are defined as either motile or non-motile Gram-negative non-spore forming bacilli that possess β-galactosidase to produce acids and gases under their optimal growth temperature of 35–37 °C. They can be aerobes or facultative aerobes, and are a commonly used indicator of low sanitary quality of foods, milk, and water. Coliforms can be found in the aquatic environment, in soil and on vegetation; they are universally present in large numbers in the feces of warm-blooded animals as they are known to inhabit the gastrointestinal system. While coliform bacteria are not normally causes of serious illness, they are easy to culture, and their presence is used to infer that other pathogenic organisms of fecal origin may be present in a sample, or that said sample is not safe to consume. Such pathogens include disease-causing bacteria, viruses, or protozoa and many multicellular parasites. Every drinking water source must be tested for the presence of these total coliform bacteria.
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.
MacConkey agar is a selective and differential culture medium for bacteria. It is designed to selectively isolate Gram-negative and enteric bacteria and differentiate them based on lactose fermentation. 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.
Bacillary dysentery is a type of dysentery, and is a severe form of shigellosis. It is associated with species of bacteria from the family Enterobacteriaceae. The term is usually restricted to Shigella infections.
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.
Simmons' citrate agar is used for differentiating gram-negative bacteria on the basis of citrate utilization, especially for distinguishing Gammaproteobacteria of the family Enterobacteriaceae or even between species of the same genus. For example, Salmonella enteritidis would yield a positive (blue) result on Simmons’ agar and thus be distinguished from other Salmonella species like Salmonella typhi, Salmonella pullorum, and Salmonella gallinarum, which would yield a negative (green) result.
Bile Esculin Agar (BEA) is a selective differential agar used to isolate and identify members of the genus Enterococcus, formerly part of the "group D streptococci".
XLT Agar is a selective culture medium for the isolation and identification of salmonellae from food and environmental samples. It is similar to XLD agar; however, the agar is supplemented with the surfactant, Tergitol 4, which causes inhibition of Proteus spp. and other non-Salmonellae. Successful growth of Salmonella will result in growth of red colonies with a black centre.
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.
Buffered charcoal yeast extract (BCYE) agar is a selective growth medium used to culture or grow certain types of bacteria, particularly the Gram-negative species Legionella pneumophila. It has also been used for the laboratory diagnosis of Acanthamoeba keratitis, Francisella and Nocardia spp. It contains L-cysteine amino acid and ferric pyrophosphate that assist in the growth of Legionnaire's species. The charcoal within the medium acts as a detoxicant because it decomposes hydrogen peroxide which is toxic to the legionellae. The yeast extract in BCYE is the rich source of nutrients (vitamins, nitrogen, and carbon) that the bacteria depends on for growth. BCYE also has ACES buffer which maintains an optimal pH level for the bacteria to grow which is around 6.9. BCYE may be supplemented with antibiotics to select for legionellae, especially if screening an environmental or non-potable water specimen.
Pathogenic bacteria are bacteria that can cause disease. This article focuses on the bacteria that are pathogenic to humans. Most species of bacteria are harmless and are often beneficial but others can cause infectious diseases. The number of these pathogenic species in humans is estimated to be fewer than a hundred. By contrast, several thousand species are part of the gut flora present in the digestive tract.
Hektoen enteric agar is a selective and differential agar 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.
Sabouraud agar or Sabouraud dextrose agar (SDA) is a type of agar growth medium containing peptones. It is used to cultivate dermatophytes and other types of fungi, and can also grow filamentous bacteria such as Nocardia. It has utility for research and clinical care.
Apocholate citrate agar (ACA) is a selective environment used to isolate Shigella and Salmonella bacteria. The name derives from apocholate and citrate in agar.
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.
The NYC medium or GC medium agar is used for isolating Gonococci.
In microbiology, the term isolation refers to the separation of a strain from a natural, mixed population of living microbes, as present in the environment, for example in water or soil, or from living beings with skin flora, oral flora or gut flora, in order to identify the microbe(s) of interest. Historically, the laboratory techniques of isolation first developed in the field of bacteriology and parasitology, before those in virology during the 20th century.
Welton Ivan Taylor was an American microbiologist, inventor and civil rights activist. He is known for his work on food-borne pathogens, notably for developing tests for Salmonella and for inventing the XLD agar, which can be used to isolate Salmonella and Shigella bacteria.