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Isolation (microbiology)
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 flora, 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 (during the 19th century), before those in virology during the 20th century
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Microbe
A microorganism, or microbe,[a] is a microscopic organism, which may exist in its single-celled form or in a colony of cells. The possible existence of unseen microbial life was suspected from ancient times, such as in Jain scriptures
Jain scriptures
from 6th-century-BC India and the 1st-century-BC book On Agriculture
Agriculture
by Marcus Terentius Varro. Microbiology, the scientific study of microorganisms, began with their observation under the microscope in the 1670s by Antonie van Leeuwenhoek. In the 1850s, Louis Pasteur
Louis Pasteur
found that microorganisms caused food spoilage, debunking the theory of spontaneous generation. In the 1880s Robert Koch
Robert Koch
discovered that microorganisms caused the diseases tuberculosis, cholera and anthrax. Microorganisms include all unicellular organisms and so are extremely diverse
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Buffered Charcoal Yeast Extract Agar
Buffered charcoal yeast extract (BCYE) agar is a selective growth medium used to culture or grow certain bacteria, particularly the Gram-negative
Gram-negative
species Legionella pneumophila.[1] It has also been used for the recovery of Acanthamoeba keratitis.[2] References[edit]^ "Archived copy" (PDF). Archived from the original (PDF) on 2012-06-12. Retrieved 2012-05-11.  ^ Penland RL, Wilhelmus KR (October 1998). "Laboratory diagnosis of Acanthamoeba keratitis
Acanthamoeba keratitis
using buffered charcoal-yeast extract agar". Am. J. Ophthalmol. 126 (4): 590–2. doi:10.1016/S0002-9394(98)00125-1
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Phenol Red
Phenol
Phenol
red (also known as phenolsulfonphthalein or PSP) is a pH indicator frequently used in cell biology laboratories.Contents1 Chemical structure and properties 2 Phenolsulfonphthalein test 3 Indicator for cell cultures 4 Estrogen
Estrogen
mimic 5 Use in swimming pool test kits 6 References 7 External linksChemical structure and properties[edit] Phenol
Phenol
red exists as a red crystal that is stable in air. Its solubility is 0.77 grams per liter (g/l) in water and 2.9 g/l in ethanol.[1] It is a weak acid with pKa = 8.00 at 20 °C (68 °F). A solution of phenol red is used as a pH indicator, often in cell culture. Its color exhibits a gradual transition from yellow (λmax = 443 nm[2]) to red (λmax = 570 nm[3]) over the pH range 6.8 to 8.2
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PH Indicator
A pH indicator is a halochromic chemical compound added in small amounts to a solution so the pH (acidity or basicity) of the solution can be determined visually. Hence, a pH indicator is a chemical detector for hydronium ions (H3O+) or hydrogen ions (H+) in the Arrhenius model. Normally, the indicator causes the color of the solution to change depending on the pH. Indicators can also show change in other physical properties; for example, olfactory indicators show change in their odor. The pH value of a neutral solution is 7.0. Solutions with a pH value below 7.0 are considered acidic and solutions with pH value above 7.0 are basic (alkaline). As most naturally occurring organic compounds are weak protolytes, carboxylic acids and amines, pH indicators find many applications in biology and analytical chemistry. Moreover, pH indicators form one of the three main types of indicator compounds used in chemical analysis
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Mannitol
Mannitol
Mannitol
is a type of sugar alcohol which is also used as a medication.[1][2] As a sugar, it is often used as a sweetener in diabetic food, as it is poorly absorbed from the intestines.[1] As a medication, it is used to decrease pressure in the eyes, as in glaucoma, and to lower increased intracranial pressure.[3][2] Medically, it is given by injection.[4] Effects typically begin within 15 minutes and last up to 8 hours.[4] Common side effects from medical use include electrolyte problems and dehydration.[4] Other serious side effects may include worsening heart failure and kidney problems.[4][2] It is unc
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Granada Medium
Granada medium
Granada medium
is a selective and differential culture medium designed to selectively isolate Streptococcus agalactiae
Streptococcus agalactiae
(Group B streptococcus, GBS) and differentiate it from other microorganisms. Granada Medium was developped by Dr. Manuel Rosa-Fraile et al
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Group B Streptococcus
Streptococcus
Streptococcus
agalactiae (also known as group B streptococcus or GBS) is a gram-positive coccus (round bacterium) with a tendency to form chains (as reflected by the genus name Streptococcus). It is a beta-hemolytic, catalase-negative, and facultative anaerobe.[1][2] In general, GBS is a harmless commensal bacterium being part of the human microbiota colonizing the gastrointestinal and genitourinary tract of up to 30% of healthy human adults (asymptomatic carriers). Nevertheless, GBS can cause severe invasive infections.[3] Streptococcus
Streptococcus
agalactiae is the species designation for streptococci belonging to group B of the Lancefield classification. GBS is surrounded by a bacterial capsule composed of polysaccharides (exopolysacharide)
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Bacterial Colony
In biology, a colony is composed of two or more conspecific individuals living in close association with, or connected to, one another. This association is usually for mutual benefit such as stronger defense or the ability to attack bigger prey.[1] It is a cluster of identical cells (clones) on the surface of (or within) a solid medium, usually derived from a single parent cell, as in bacterial colony.[2] In contrast, a solitary organism is one in which all individuals live independently and have all of the functions needed to survive and reproduce. Colonies, in the context of development, may be composed of two or more unitary (or solitary) organisms or be modular organisms. Unitary organisms have determinate development (set life stages) from zygote to adult form and individuals or groups of individuals (colonies) are visually distinct
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Blood Agar
An agar plate is a Petri dish
Petri dish
that contains a solid growth medium, typically agar plus nutrients, used to culture small organisms such as microorganisms. Sometimes selective compounds are added to influence growth, such as antibiotics.[1]96 pinner used to perform spot assays with yeast, fungal or bacterial cellsIndividual microorganisms placed on the plate will grow into individual colonies, each a clone genetically identical to the individual ancestor organism (except for the low, unavoidable rate of mutation). Thus, the plate can be used either to estimate the concentration of organisms in a liquid culture or a suitable dilution of that culture using a colony counter, or to generate genetically pure cultures from a mixed culture of genetically different organisms. Several methods are available to plate out cells. One technique is known as "streaking"
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Hemolysis
Hemolysis
Hemolysis
or haemolysis, also known by several other names, is the rupturing (lysis) of red blood cells (erythrocytes) and the release of their contents (cytoplasm) into surrounding fluid (e.g. blood plasma). Hemolysis
Hemolysis
may occur in vivo or in vitro (inside or outside the body). Hemolysins damage the host cytoplasmic membrane, causing cell lysis and death. The activity of these toxins is most easily observed with assays involving the lysis of red blood cells (erythrocytes). Some hemolysins attack the phospholipid of the host cytoplasmic membrane. Because the phospholipid lecithin (phosphatidylcholine) is often used as a substrate, these enzymes are called lecithinases or phospholipases
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Legionella Species
Legionella adelaidensis[1] Legionella anisa[1] Legionella beliardensis[1] Legionella birminghamensis[1] Legionella bozemanae[1] Legionella brunensis[1] Legionella busanensis[1] Legionella cardiaca[1] Legionella cherrii[1] Legionella cincinnatiensis[1] Legionella clemsonensis[2] Legionella donaldsonii[1] Legionella drancourtii[1] Legionella dresdenensis[1] Legionella drozanskii[1] Legionella dumoffii[1] Legionella erythra[1] Legionella fairfieldensis[1] Legionella fallonii[1] Legionella feeleii[1] Legionella geestiana[1] Legionella genomospecies 1 Legionella gormanii[1] Legionella gratiana[1] Legionella gresilensis[1] Legionella hackeliae[1] Legionella impletisoli[1] Legionella israelensis[1] Legionella jamestowniensis[1] Candidatus Legionella jeonii Legionella jordanis[1] Legionella lansingensis[1] Legionella londiniensis[1] Legionella longbeachae[1] Legionella lytica[1] Legionella maceachernii[1] Legionella massiliensis[1] Legionella micdadei[1] Legionella monrovica[1] Legionella mora
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Charcoal
Charcoal
Charcoal
is the lightweight black carbon and ash residue hydrocarbon produced by removing water and other volatile constituents from animal and vegetation substances. Charcoal
Charcoal
is usually produced by slow pyrolysis — the heating of wood or other substances in the absence of oxygen (see char and biochar)
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Chocolate Agar
Chocolate
Chocolate
agar (CHOC) or chocolate blood agar (CBA) – is a nonselective, enriched growth medium used for isolation of pathogenic bacteria.[1][2][3] It is a variant of the blood agar plate, containing red blood cells that have been lysed by slowly heating to 80 °C. Chocolate
Chocolate
agar is used for growing fastidious respiratory bacteria, such as Haemophilus influenzae
Haemophilus influenzae
and Neisseria
Neisseria
meningitidis.[4] In addition, some of these bacteria, most notably H. influenzae, need growth factors such as NAD (factor V) and hemin (factor X), which are inside red blood cells; thus, a prerequisite to growth for these bacteria is lysis of the red blood cells. The heat also inactivates enzymes which could otherwise degrade NAD
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Mannitol Salt Agar
Mannitol
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 by distinguishing pathogenic microbes in a short period of time.[1] It contains a high concentration (about 7.5%-10%) of salt (NaCl), making it selective for Gram-positive bacteria
Gram-positive bacteria
( Staphylococcus
Staphylococcus
and Micrococcaceae) since this level of salt is inhibitory to most other bacteria.[2] 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
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Thioglycolate Broth
Thioglycolate broth is a multipurpose, enriched, differential medium used primarily to determine the oxygen requirements of microorganisms. Sodium thioglycolate in the medium consumes oxygen and permits the growth of obligate anaerobes.[1] This, combined with the diffusion of oxygen from the top of the broth, produces a range of oxygen concentrations in the medium along its depth. The oxygen concentration at a given level is indicated by a redox-sensitive dye such as resazurine that turns pink in the presence of oxygen.Thioglycolate broth medium is recommended to isolate strict anaerobes should an anaerobic infection be suspected.[2]This allows the differentiation of obligate aerobes, obligate anaerobes, facultative anaerobes, microaerophiles, and aerotolerant organisms
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