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Acid-fast
Acid-fastness is a physical property of certain bacterial and eukaryotic cells, as well as some sub-cellular structures, specifically their resistance to decolorization by acids during laboratory staining procedures. Once stained as part of a sample, these organisms can resist the acid and/or ethanol-based decolorization procedures common in many staining protocols, hence the name ''acid-fast''. The mechanisms of acid-fastness vary by species, although the most well-known example is in the genus '' Mycobacterium'', which includes the species responsible for tuberculosis and leprosy. The acid-fastness of ''Mycobacteria'' is due to the high mycolic acid content of their cell walls, which is responsible for the staining pattern of poor absorption followed by high retention. Some bacteria may also be partially acid-fast, such as '' Nocardia''. Acid-fast organisms are difficult to characterize using standard microbiological techniques, though they can be stained using concentrated dye ...
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Ziehl–Neelsen Stain
Ziehl–Neelsen staining is a type of acid-fast stain, first introduced by Paul Ehrlich. Ziehl–Neelsen staining is a bacteriological stain used to identify acid-fast organisms, mainly Mycobacteria. It is named for two German doctors who modified the stain: the bacteriologist Franz Ziehl (1859–1926) and the pathologist Friedrich Neelsen (1854–1898). Mycobacteria In anatomic pathology specimens, immunohistochemistry and modifications of Ziehl–Neelsen staining (such as the Fite-Faraco method) have comparable diagnostic utility. Both of them are superior to traditional Ziehl–Neelsen staining. The genus ''Mycobacterium'' is a slow growing bacteria, made up of small rods that are slightly curved or straight, and are considered to be gram positive. Some mycobacteria are free-living saprophytes, but many are pathogens that cause disease in animals and humans. ''Mycobacterium bovis'' causes tuberculosis in cattle. Since tuberculosis can be spread to humans, milk is pasteurized ...
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Kinyoun Stain
The Kinyoun method or Kinyoun stain (cold method), developed by Joseph J. Kinyoun, is a procedure used to stain acid-fast species of the bacterial genus ''Mycobacterium''. It is a variation of a method developed by Robert Koch in 1882. Certain species of bacteria have a waxy lipid called mycolic acid, in their cell walls which allow them to be stained with Acid-Fast better than a Gram-Stain. The unique ability of mycobacteria to resist decolorization by acid-alcohol is why they are termed acid-fast. It involves the application of a primary stain (basic fuchsin), a decolorizer (acid-alcohol), and a counterstain (methylene blue). Unlike the Ziehl–Neelsen stain Ziehl–Neelsen staining is a type of acid-fast stain, first introduced by Paul Ehrlich. Ziehl–Neelsen staining is a bacteriological stain used to identify acid-fast organisms, mainly Mycobacteria. It is named for two German doctors who modif ... (Z-N stain), the Kinyoun method of staining does not require he ...
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Tuberculosis
Tuberculosis (TB) is an infectious disease usually caused by '' Mycobacterium tuberculosis'' (MTB) bacteria. Tuberculosis generally affects the lungs, but it can also affect other parts of the body. Most infections show no symptoms, in which case it is known as latent tuberculosis. Around 10% of latent infections progress to active disease which, if left untreated, kill about half of those affected. Typical symptoms of active TB are chronic cough with blood-containing mucus, fever, night sweats, and weight loss. It was historically referred to as consumption due to the weight loss associated with the disease. Infection of other organs can cause a wide range of symptoms. Tuberculosis is spread from one person to the next through the air when people who have active TB in their lungs cough, spit, speak, or sneeze. People with Latent TB do not spread the disease. Active infection occurs more often in people with HIV/AIDS and in those who smoke. Diagnosis of active TB is ...
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Carbol Fuchsin
Carbol fuchsin, carbol-fuchsin, carbolfuchsin, or Castellani's paint ( CAS ) is a mixture of phenol and basic fuchsin that is used in bacterial staining procedures. It is commonly used in the staining of mycobacteria because it has an affinity for the mycolic acids found in their cell membranes. It is a component of Ziehl–Neelsen stain, a differential stain. Carbol fuchsin is used as the primary stain dye to detect acid-fast bacteria because it is more soluble in the cells' wall lipids than in the acid alcohol. If the bacteria is acid-fast the bacteria will retain the initial red color of the dye because they are able to resist the destaining by acid alcohol (0.4–1% HCl in 70% EtOH). Additionally, it can be used for the staining of bacterial spores. Carbol-fuchsin is also used as a topical antiseptic An antiseptic (from Greek ἀντί ''anti'', "against" and σηπτικός ''sēptikos'', "putrefactive") is an antimicrobial substance or compound that is applied to livi ...
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Leprosy
Leprosy, also known as Hansen's disease (HD), is a long-term infection by the bacteria ''Mycobacterium leprae'' or ''Mycobacterium lepromatosis''. Infection can lead to damage of the nerves, respiratory tract, skin, and eyes. This nerve damage may result in a lack of ability to feel pain, which can lead to the loss of parts of a person's extremities from repeated injuries or infection through unnoticed wounds. An infected person may also experience muscle weakness and poor eyesight. Leprosy symptoms may begin within one year, but, for some people, symptoms may take 20 years or more to occur. Leprosy is spread between people, although extensive contact is necessary. Leprosy has a low pathogenicity, and 95% of people who contract ''M. leprae'' do not develop the disease. Spread is thought to occur through a cough or contact with fluid from the nose of a person infected by leprosy. Genetic factors and immune function play a role in how easily a person catches the disease. Lepro ...
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Nocardia
''Nocardia'' is a genus of weakly staining Gram-positive, catalase-positive, rod-shaped bacteria. It forms partially acid-fast beaded branching filaments (acting as fungi, but being truly bacteria). It contains a total of 85 species. Some species are nonpathogenic, while others are responsible for nocardiosis. ''Nocardia'' species are found worldwide in soil rich in organic matter. In addition, they are oral microflora found in healthy gingiva, as well as periodontal pockets. Most ''Nocardia'' infections are acquired by inhalation of the bacteria or through traumatic introduction. Culture and staining ''Nocardia'' colonies have a variable appearance, but most species appear to have aerial hyphae when viewed with a dissecting microscope, particularly when they have been grown on nutritionally limiting media. ''Nocardia'' grow slowly on nonselective culture media, and are strict aerobes with the ability to grow in a wide temperature range. Some species are partially acid-fast ...
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Mycobacterium Tuberculosis Ziehl-Neelsen Stain 640
''Mycobacterium'' is a genus of over 190 species in the phylum Actinomycetota, assigned its own family, Mycobacteriaceae. This genus includes pathogens known to cause serious diseases in mammals, including tuberculosis ('' M. tuberculosis'') and leprosy ('' M. leprae'') in humans. The Greek prefix ''myco-'' means 'fungus', alluding to this genus' mold-like colony surfaces. Since this genus has cell walls with Gram-positive and Gram-negative features, acid-fast staining is used to emphasize their resistance to acids, compared to other cell types. Metabolism and Morphology Mycobacteria are aerobic with 0.2-0.6 µm wide and 1.0-10 µm long rod shapes. They are generally non-motile, except for the species ''Mycobacterium marinum'', which has been shown to be motile within macrophages. Mycobacteria possess capsules and most do not form endospores. ''M. marinum'' and perhaps ''M. bovis'' have been shown to sporulate; however, this has been contested by further research. The disti ...
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Gram Stain
In microbiology and bacteriology, Gram stain (Gram staining or Gram's method), is a method of staining used to classify bacterial species into two large groups: gram-positive bacteria and gram-negative bacteria. The name comes from the Danish bacteriologist Hans Christian Gram, who developed the technique in 1884. Gram staining differentiates bacteria by the chemical and physical properties of their cell walls. Gram-positive cells have a thick layer of peptidoglycan in the cell wall that retains the primary stain, crystal violet. Gram-negative cells have a thinner peptidoglycan layer that allows the crystal violet to wash out on addition of ethanol. They are stained pink or red by the counterstain, commonly safranin or fuchsine. Lugol's iodine solution is always added after addition of crystal violet to strengthen the bonds of the stain with the cell membrane. Gram staining is almost always the first step in the preliminary identification of a bacterial organism. While Gram s ...
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Mycobacterium
''Mycobacterium'' is a genus of over 190 species in the phylum Actinomycetota, assigned its own family, Mycobacteriaceae. This genus includes pathogens known to cause serious diseases in mammals, including tuberculosis ('' M. tuberculosis'') and leprosy ('' M. leprae'') in humans. The Greek prefix ''myco-'' means 'fungus', alluding to this genus' mold-like colony surfaces. Since this genus has cell walls with Gram-positive and Gram-negative features, acid-fast staining is used to emphasize their resistance to acids, compared to other cell types. Metabolism and Morphology Mycobacteria are aerobic with 0.2-0.6 µm wide and 1.0-10 µm long rod shapes. They are generally non-motile, except for the species ''Mycobacterium marinum'', which has been shown to be motile within macrophages. Mycobacteria possess capsules and most do not form endospores. ''M. marinum'' and perhaps ''M. bovis'' have been shown to sporulate; however, this has been contested by further research. The disti ...
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Bacteria
Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, 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 Earth's crust. Bacteria are vital in many stages of the nutrient cycle by recycling nutrients such as the fixation of nitrogen from the atmosphere. The nutrient cycle includes the decomposition of dead bodies; bacteria are responsible for the putrefaction stage in this process. In the biological communities surrounding hydrothermal vents and cold seeps, extremophile bacteria provide the nutrients needed to sustain life by converting dissolved compounds, such as hydrogen sulphide and methane, to energy. Bacteria also live in symbiotic and parasitic relationsh ...
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Auramine Phenol Stain
Auramine phenol stain is a stain used in clinical microbiology and histology to identify tuberculosis mycobacteria. There are two types of auramine phenol stains, 1 and 2 to stain mycobacterium species and cryptosporidium respectively. Both are fluorescent Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation. It is a form of luminescence. In most cases, the emitted light has a longer wavelength, and therefore a lower photon energy, ... stains. The bacteria or the parasites appear brilliant greenish yellow against dark background. Mycolic acids of the mycobacteria keep this stain when decolorising with the acid alcohol. The method is more rapid and sensitive than ZN technique. Method * Smears are prepared just like that for ZN staining * Stain with Auramine-Phenol for 20 mins * Rinse with water * Decolourise in acid alcohol * Rinse with water * Counterstain with 0.1% potassium permanganate for 30 seconds * Rins ...
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Mycolic Acid
Mycolic acids are long fatty acids found in the cell walls of the Mycolata taxon, a group of bacteria that includes ''Mycobacterium tuberculosis'', the causative agent of the disease tuberculosis. They form the major component of the cell wall of mycolata species. Despite their name, mycolic acids have no biological link to fungi; the name arises from the filamentous appearance their presence gives mycolata under high magnification. The presence of mycolic acids in the cell wall also gives mycolata a distinct gross morphological trait known as " cording". Mycolic acids were first isolated by Stodola ''et al.'' in 1938 from an extract of ''M. tuberculosis''. Mycolic acids are composed of a longer beta-hydroxy chain with a shorter alpha-alkyl side chain. Each molecule contains between 60 and 90 carbon atoms. The exact number of carbons varies by species and can be used as an identification aid. Most mycolic acids also contain various functional groups. Mycolic acids of ''M. ...
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