MacConkey agar is an indicator, a selective and differential culture medium for bacteria designed to selectively isolate Gram-negative and enteric (normally found in the intestinal tract) bacilli and differentiate them based on lactose fermentation.The crystal violet and bile salts inhibit the growth of Gram-positive organisms which allows for the selection and isolation of gram-negative bacteria. Enteric bacteria that have the ability to ferment lactose can be detected using the carbohydrate lactose, and the pH indicator neutral red.
A microbiological culture, or microbial culture, is a method of multiplying microbial organisms by letting them reproduce in predetermined culture medium under controlled laboratory conditions. Microbial cultures are foundational and basic diagnostic methods used as a research tool in molecular biology.
Bacteria are a type of biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, bacteria have a number of shapes, ranging from spheres to rods and spirals. Bacteria were among the first life forms to appear on Earth, and are present in most of its habitats. Bacteria inhabit soil, water, acidic hot springs, radioactive waste, and the deep biosphere of the earth's crust. Bacteria also live in symbiotic and parasitic relationships with plants and animals. Most bacteria have not been characterised, and only about 27 percent of the bacterial phyla have species that can be grown in the laboratory . The study of bacteria is known as bacteriology, a branch of microbiology.
A bacillus or bacilliform bacterium is a rod-shaped bacterium or archaeon. Bacilli are found in many different taxonomic groups of bacteria. However, the name Bacillus capitalized and italicized, refers to a specific genus of bacteria. The name Bacilli, capitalized but not italicized, can also refer to a less specific taxonomic group of bacteria that includes two orders, one of which contains the genus Bacillus. When the word is formatted with lowercase and not italicized, 'bacillus', it will most likely be referring to shape and not to the genus at all. Bacilliform bacteria are also often simply called rods when the bacteriologic context is clear. Sea Bacilli usually divide in the same plane and are solitary, but can combine to form diplobacilli, streptobacilli, and palisades.
It contains bile salts (to inhibit most Gram-positive bacteria), crystal violet dye (which also inhibits certain Gram-positive bacteria), neutral red dye (which turns pink if the microbes are fermenting lactose).
Crystal violet or gentian violet is a triarylmethane dye used as a histological stain and in Gram's method of classifying bacteria. Crystal violet has antibacterial, antifungal, and anthelmintic properties and was formerly important as a topical antiseptic. The medical use of the dye has been largely superseded by more modern drugs, although it is still listed by the World Health Organization.
Neutral red is a eurhodin dye used for staining in histology. It stains lysosomes red. It is used as a general stain in histology, as a counterstain in combination with other dyes, and for many staining methods. Together with Janus Green B, it is used to stain embryonal tissues and supravital staining of blood. Can be used for staining Golgi apparatus in cells and Nissl granules in neurons.
Lactose is a disaccharide. It is a sugar composed of galactose and glucose subunits and has the molecular formula C12H22O11. Lactose makes up around 2–8% of milk (by weight). The name comes from lac (gen. lactis), the Latin word for milk, plus the suffix -ose used to name sugars. The compound is a white, water-soluble, non-hygroscopic solid with a mildly sweet taste. It is used in the food industry.
Sodium chloride, commonly known as salt, is an ionic compound with the chemical formula NaCl, representing a 1:1 ratio of sodium and chloride ions. With molar masses of 22.99 and 35.45 g/mol respectively, 100 g of NaCl contains 39.34 g Na and 60.66 g Cl. Sodium chloride is the salt most responsible for the salinity of seawater and of the extracellular fluid of many multicellular organisms. In its edible form of table salt, it is commonly used as a condiment and food preservative. Large quantities of sodium chloride are used in many industrial processes, and it is a major source of sodium and chlorine compounds used as feedstocks for further chemical syntheses. A second major application of sodium chloride is de-icing of roadways in sub-freezing weather.
Agar or agar-agar, is a jelly-like substance, obtained from red algae.
Water is a transparent, tasteless, odorless, and nearly colorless chemical substance, which is the main constituent of Earth's streams, lakes, and oceans, and the fluids of most living organisms. It is vital for all known forms of life, even though it provides no calories or organic nutrients. Its chemical formula is H2O, meaning that each of its molecules contains one oxygen and two hydrogen atoms, connected by covalent bonds. Water is the name of the liquid state of H2O at standard ambient temperature and pressure. It forms precipitation in the form of rain and aerosols in the form of fog. Clouds are formed from suspended droplets of water and ice, its solid state. When finely divided, crystalline ice may precipitate in the form of snow. The gaseous state of water is steam or water vapor. Water moves continually through the water cycle of evaporation, transpiration (evapotranspiration), condensation, precipitation, and runoff, usually reaching the sea.
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.
The medium was developed by Alfred Theodore MacConkey while working as a bacteriologist for the Royal Commission on Sewage Disposal.
Alfred Theodore MacConkey (1861-1931) was the British bacteriologist who developed MacConkey's agar, a selective medium that is used in the diagnosis of enteric pathogens. He was born McConkey but appears to have spelled his name "MacConkey" from at least 1881 and in all his published papers
Bacteriology is the branch and specialty of biology that studies the morphology, ecology, genetics and biochemistry of bacteria as well as many other aspects related to them. This subdivision of microbiology involves the identification, classification, and characterization of bacterial species. Because of the similarity of thinking and working with microorganisms other than bacteria, such as protozoa, fungi, and viruses, there has been a tendency for the field of bacteriology to extend as microbiology. The terms were formerly often used interchangeably. However, bacteriology can be classified as a distinct science.
The Royal Commission on Sewage Disposal was established by the British government in 1898 to report on:
(1) What method or methods of treating and disposing of sewage may properly be adopted,consistently with due regard for the requirements of the existing law, for the protection of the public health, and for the economical and efficient discharge of the duties of local, authorities ; and
(2) If more than one method may be so adopted, by what rules, in relation to the nature or volume of sewage, or the population to be served, or other varying circumstances or requirements, should the particular method of treatment and disposal to be adopted be determined
Using neutral red pH indicator, the agar distinguishes those Gram-negative bacteria that can ferment the sugar lactose (Lac+) from those that cannot (Lac-).
In chemistry, pH is a scale used to specify how acidic or basic a water-based solution is. Acidic solutions have a lower pH, while basic solutions have a higher pH. At room temperature (25 °C), pure water is neither acidic nor basic and has a pH of 7.
This medium is also known as an "indicator medium" and a "low selective medium". Presence of bile salts inhibits swarming by Proteus species.
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 neighbourhood of the colony, causing the medium surrounding the colony to become hazy.
Examples of non-lactose fermenting bacteria are Salmonella , Proteus species, Yersinia , Pseudomonas aeruginosa and Shigella .
Some organisms ferment lactose slowly or weakly, and are sometimes put in their own category. These include Serratiaand Citrobacter .
Some organisms, especially Klebsiella and Enterobacter, produce mucoid colonies which appear very moist and sticky. This phenomenon happens because the organism is producing a capsule, which is predominantly made from the lactose sugar in the agar.
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.
The family Enterobacteriaceae is a large family of Gram-negative bacteria. It was first proposed by Rahn in 1936, and now includes over 50 genera and more than 200 species. Its classification above the level of family is still a subject of debate, but one classification places it in the order Enterobacteriales of the class Gammaproteobacteria in the phylum Proteobacteria.
An agar plate is a Petri dish that contains agar as a solid growth medium plus nutrients, 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.
Proteus vulgaris is a rod-shaped, nitrate-reducing, indole+ and catalase-positive, hydrogen sulfide-producing, Gram-negative bacterium that inhabits the intestinal tracts of humans and animals. It can be found in soil, water, and fecal matter. It is grouped with the Enterobacteriaceae and is an opportunistic pathogen of humans. It is known to cause wound infections and other species of its genera are known to cause urinary tract infections.
Proteus mirabilis is a Gram-negative, facultatively anaerobic, rod-shaped bacterium. It shows swarming motility and urease activity. P. mirabilis causes 90% of all Proteus infections in humans. It is widely distributed in soil and water.
Coliform bacteria are defined as rod-shaped Gram-negative non-spore forming and motile or non-motile bacteria which can ferment lactose with the production of acid and gas when incubated at 35–37°C. Due to the limited ability of certain coliform bacteria to ferment lactose, the definition has changed to bacteria containing the enzyme B-galactosidase. They are a commonly used indicator of sanitary quality of foods 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. While coliforms themselves are not normally causes of serious illness, they are easy to culture, and their presence is used to indicate that other pathogenic organisms of fecal origin may be present. Such pathogens include disease-causing bacteria, viruses, or protozoa and many multicellular parasites. Coliform procedures are performed in aerobic or anaerobic conditions.
Eosin methylene blue is a selective stain for Gram-negative bacteria. EMB contains dyes that are toxic to Gram-positive bacteria. EMB is the selective and differential medium for coliforms. It is a blend of two stains, eosin and methylene blue in the ratio of 6:1. A common application of this stain is in the preparation of EMB agar, a differential microbiological medium, which slightly inhibits the growth of Gram-positive bacteria and provides a color indicator distinguishing between organisms that ferment lactose and those that do not. Organisms that ferment lactose display "nucleated colonies"—colonies with dark centers.
A growth medium or culture medium is a solid, liquid or semi-solid designed to support the growth of microorganisms or cells, or small plants like the moss Physcomitrella patens. Different types of media are used for growing different types of cells.
A fecal coliform is a facultatively anaerobic, rod-shaped, gram-negative, non-sporulating bacterium. Coliform bacteria generally originate in the intestines of warm-blooded animals. Fecal coliforms are capable of growth in the presence of bile salts or similar surface agents, are oxidase negative, and produce acid and gas from lactose within 48 hours at 44 ± 0.5°C. The term "thermotolerant coliform" is more correct and is gaining acceptance over "fecal coliform".
Burkholderia cepacia complex (BCC), or simply Burkholderia cepacia, is a group of catalase-producing, lactose-nonfermenting, Gram-negative bacteria composed of at least 20 different species, including B. cepacia, B. multivorans, B. cenocepacia, B. vietnamiensis, B. stabilis, B. ambifaria, B. dolosa, B. anthina, B. pyrrocinia and B. ubonensis. B. cepacia is an opportunistic human pathogen that most often causes pneumonia in immunocompromised individuals with underlying lung disease. Patients with sickle-cell haemoglobinopathies are also at risk. The species also attacks young onion and tobacco plants, as well as displaying a remarkable ability to digest oil.
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.
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. This medium is important in medical laboratories as one method of distinguishing pathogenic microbes in a short period of time. It contains a high concentration of salt (NaCl), making it selective for Gram-positive bacteria since this level of salt is inhibitory to most other bacteria. It is also a differential medium for mannitol-fermenting staphylococci, containing carbohydrate 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.
De Man, Rogosa and Sharpe agar, often abbreviated to MRS, is a selective culture medium designed to favour the luxuriant growth of Lactobacilli for lab study. Developed in 1960, this medium was named for its inventors. It contains sodium acetate, which suppresses the growth of many competing bacteria. This medium has a clear brown colour.
Thayer-Martin agar is a Mueller-Hinton agar with 5% chocolate sheep blood and antibiotics. It is used for culturing and primarily isolating pathogenic Neisseria bacteria, including Neisseria gonorrhoeae and Neisseria meningitidis, as the medium inhibits the growth of most other microorganisms. When growing Neisseria meningitidis, one usually starts with a normally sterile body fluid, so a plain chocolate agar is used. Thayer-Martin agar was initially developed in 1964, with an improved formulation published in 1966.
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.
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 sp..
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. 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 N.Y.C medium or GC medium agar is used for isolating Gonococci.