Dehydrated Culture Media
Dehydrated Culture Media products are formulated to supply the required nutrients to allow for the growth of microorganisms. Used in combination with a variety of selective agents and incubation conditions a wide range of specific organisms can be isolated. With careful raw material selection of the various media components E&O can ensure a consistent level of quality and performance. For each formulation the necessary ingredients are accurately weighed, combined and blended together to produce a homogenous powdered product.
View Our Dehydrated Culture Media
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Alkaline peptone water is generally used as an enrichment medium in the isolation of Vibrio spp. from faeces. This medium may also be used for the enrichment of Vibrio spp. from food and water samples. First developed by Shread, Donovan and Lee as an enrichment broth for the growth of Aeromonas spp., Cruickshank showed that with a higher pH the medium can be used for the enrichment of Vibrio spp. The peptone is the source of the required nitrogen, carbon and vitamins. Sodium chloride maintains the osmotic balance. The high pH of the medium inhibits most enteric organisms for at least 24 hours. -
Bacillus cereus (MYP) agar is intended for the selective enumeration of Bacillus cereus in food samples. This medium utilizes two reactions namely mannitol fermentation and lecithinase production to differentiate Bacillus cereus from other related species. As B. cereus is mannitol negative the colonies are pink in colour due to the presence of the phenol red pH indicator. Lecithinase production (from the addition of egg yolk) is indicated by a white precipitate around the colonies. This medium meets the requirements of ISO 7932:2004. Beef extract and peptone provide the required carbon, nitrogen and vitamins. Sodium chloride helps maintain the osmotic balance and phenol red is the pH indicator. Mannitol is a fermentable carbohydrate. The medium is made selective by the inclusion of polymyxin B sulphate (LS0020). NB: This is a basic medium only and contains no additional supplement. It should be noted that some Proteus spp. and Gram-positive cocci may grow on this medium. Related Supplements : LS0020 Bacillus cereus Selective Supplement, BM0140 Egg Yolk Emulsion
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Bacillus cereus agar (PEMBA) is used for the selective isolation and enumeration of Bacillus cereus in food samples. It is said to be particularly suitable for the detection of small numbers of Bacillus cereus particularly in the presence of other contaminating organisms. Bacillus cereus agar (PEMBA) is based on the formulation developed by Holbrook and Anderson. The peptone provides the required carbon, nitrogen and vitamins. Mannitol is a fermentable carbohydrate. Bromothymol blue is a pH indicator used to detect mannitol fermentation. Sodium chloride maintains the osmotic balance. Magnesium sulphate provides divalent cations and sulphate. Di-sodium hydrogen phosphate and potassium dihydrogen phosphate are buffering agents. Egg yolk emulsion (BM0140) must be added to this media to assess lecithinase production. Sodium pyruvate is present to improve egg yolk precipitation and enhance B. cereus sporulation. The medium is made selective by the inclusion of polymyxin B sulphate (LS1051).
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Bacteriological peptone is an economical source of nutrients provided by a balanced mixture of meat peptones and tryptone.The growth requirements of most non fastidious organisms will be fulfilled by the range of amino acids, peptides and proteoses in this mixture.
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Baird Parker agar is a selective medium for the isolation and presumptive identification of coagulase-positive staphylococci. This medium is used extensively for detecting Staphylococcus aureus in foods, dairy products, and other materials. Enzymatic digest of casein, meat extract and yeast extract provide the required carbon, nitrogen and vitamins. The medium is made selective by the inclusion of lithium chloride and the addition of potassium tellurite. Glycine and sodium pyruvate are included as growth enhancers while the pyruvate also acts as a neutraliser of toxic peroxides. Coagulase positive staphylococci are differentiated by the addition of egg yolk tellurite (BM0530) due to their ability to break down the egg yolk. Clear zones around colonies form due to the proteolytic action of lecithinase. A secondary opaque zone, surrounding presumptive positive colonies, may also form due to lipase activity. Potassium tellurite is reduced by staphylococci giving rise to blackening of colonies. NB: Any black colonies (with halo (typical) or without the halo (atypical)) on this medium must be confirmed as Staphylococcus aureus by further tests (e.g. coagulase test or latex agglutination etc.) Related Supplements : BM0530 Egg Yolk Tellurite
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Beef extract is sourced from bovine tissue and supports the nutritive needs of microorganisms. Providing a mixture of amino acids, peptides, minerals and vitamins this product is recommended for use in microbiological culture media for the examination of water, milk and other materials.
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Bile Aesculin Medium is generally used for the differentiation and presumptive identification of Group D streptococci (enterococci). Enterococci hydrolyse aesculin forming, amongst other products, aesculetin which in turn combines with Ferric ammonium citrate producing a dark brown or black complex. The presence of Bile salts in the medium inhibits gram positive organisms other than enterococci. The medium can also be used for the presumptive identification of certain organisms within the Enterobacteriaceae group such as Klebsiella spp., Enterobacter spp., etc.
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KM0034 Blood Agar Base No. 2 is a general-purpose medium with is designed to be enriched with various concentrations of horse or sheep blood and is suitable for the isolation of most organisms including many fastidious anaerobes of clinical significance. Blood Agar Base No. 2, frequently shortened to BAB No.2, was developed to meet the demand for an especially nutritious blood agar base which would permit the maximum recovery of delicate organisms, such as streptococci, pneumococci, and other fastidious microorganisms, without interfering with their haemolytic reactions. Haemolysis observations may vary with the type of blood being used and previous studies have shown that sheep blood provides the most reliable colony and haemolysis characteristics. When the blood is chocolated the medium gives good recovery of Haemophilus species. KM0034, when prepared with required blood type, is recommended as a non-selective blood agar by UK Standards for Microbiology Investigations and is tested in accordance with the principals of ISO 11133:2014 . Related Supplements : Defibrinated Sheep Blood, Defibrinated Horse Blood, LS0008 Staph/Strep Selective Supplement, LS0017 Neomycin Selective Supplement
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Brain heart infusion agar is very nutritious general-purpose medium and is suitable for the isolation of most micro-organisms including many fastidious organisms. The formulation is a modification of that from Rosenow and Hayden. The medium is not recommended for the determination of haemolytic reactions because of the glucose content. The nitrogen, vitamin and carbon sources are supplied by the Brain-Heart infusion solids and peptone. Glucose serves as the carbohydrate source and sodium chloride aids in maintaining the osmotic balance. A phosphate buffer, disodium hydrogen phosphate, is incorporated to help neutralize any acids produced as a result of glucose utilization and thus maintain viability of the organisms. NB: Organisms that produce large amounts of acid in the medium may overwhelm the buffering system and as a result may ‘auto-sterilize’ the culture.
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Brain heart infusion broth is very nutritious isotonic medium with a low concentration of glucose to stimulate early growth. The formulation is a modification of that from Rosenow and Hayden. Brain heart infusion broth is suitable for the isolation of most micro-organisms including many fastidious organisms and, with the appropriate enrichment, is suitable as a base for blood culture medium. The nitrogen, vitamin and carbon sources are supplied by the Brain-Heart infusion solids and peptone. Glucose serves as the carbohydrate source and sodium chloride aids in maintaining the osmotic balance. A phosphate buffer, disodium hydrogen phosphate, is incorporated to help neutralize any acids produced as a result of glucose utilization and thus maintain viability of the organisms. NB: Organisms that produce large amounts of acid in the medium may overwhelm the buffering system and as a result may auto-sterilize the culture.
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Brain-heart infusion solids is a dehydrated infusion of porcine brains and heart. This product can be used in preparing microbiological culture media providing nitrogen, vitamins, minerals and amino acids.
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KM0038 Brilliant Green Agar is used for the selective isolation of Salmonella species from clinical specimens and food samples. Brilliant Green Agar was first cited by Kristensen et al. and later modified to improve selectivity. The medium is highly selective and may not be suitable for the isolation of Salmonella spp. from samples where numbers of salmonellae are low. If this is suspected to be the case, or if the sample is suspected to contain Salmonella typhi, other selective media such as Xylose Lysine Deoxycholate (XLD) Agar (E&O KM0013), Hektoen Enteric Agar (E&O KM0032) or Deoxycholate Citrate Agar (E&O KM0050) may be inoculated with the specimen in parallel. The United States Pharmacopeia indicates that the medium may be used to confirm the absence of salmonellae in nutritional and dietary supplements. The medium may also be used as a secondary plating medium for subculture from selective enrichment media during food and environmental testing, and as a primary isolation medium in the identification of Salmonella species from clinical specimens according to Public Health England’s UK Standards for Microbiology Investigations.
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Brucella broth is a non-selective medium for the cultivation of Brucella spp. and other fastidious microorganisms. Brucella broth has been developed from the APHA formulation for Albimi broth. (1&2) The enzymatic digest of casein, enzymatic digest of animal tissues and yeast extract provide the necessary carbon, nitrogen and vitamin sources for this medium. Glucose is a fermentable carbohydrate and sodium chloride maintains the osmotic balance of the medium. Sodium bisulfite is added to enhance growth. References (1) Hausler, W. J. (ed.). 1976. Standard methods for the examination of dairy products, 14th ed. American Public Health Association, Washington, D.C. (2) ISO 10272-1:2006. Microbiology of food and animal feeding stuffs – Horizontal method for detection and enumeration of Campylobacter spp.
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Brucella Medium Base is a general purpose medium for the cultivation of Brucella spp. and other fastidious microorganisms. Brucella Medium Base has been developed from the APHA formulation for Albimi broth. The peptones act as carbon, nitrogen and vitamin source in this medium. Glucose is a fermentable carbohydrate and sodium chloride maintains the osmotic balance of the medium.
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Formulated to ISO 6579, Buffered Peptone Water (BPW) is a pre-enrichment medium designed to help sub-lethally damaged Salmonella spp. recover before introducing them into a selective enrichment medium. Enzymatic digest of casein is the source of the required nitrogen, carbon and vitamins. Sodium chloride maintains the osmotic balance in the medium. Di-sodium hydrogen phosphate and potassium di-hydrogen phosphate act as buffers in the medium. This nutritious medium is free from inhibitors and is well buffered to maintain pH 7.0 for the incubation period. Sub-lethal injury to Salmonella spp. occurs in many food processes and this pre-enrichment step greatly increases the chance of their recovery, especially if a low number of cells are present in a sample.
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Burkholderia cepacia agar base is a selective medium for the detection and isolation of Burkholderia cepacia from cystic fibrosis (CF) patients. This is an important opportunistic pathogen in CF patients and can lead to fatal infection in approximately 20% individuals that have been colonised with B. cepacia complex organisms. This medium is based on the PC medium described by Gilligan et al. Magnesium sulphate, ammonium sulphate and ferrous ammonium sulphate supports the growth of B. cepacia. Potassium di-hydrogen phosphate and di-sodium hydrogen phosphate are buffering agents, used to maintain the pH the medium. The phenol red is used as a pH indicator. If the sodium pyruvate in the medium is metabolised by B. cepacia alkaline by-products are produced which raises the pH. This causes the colour of the medium to turn pink/red around sections of heavy growth on the medium. Bile salts and crystal violet are selective agents. The associated selective supplement for this medium, LS0125, contains ticarcillin and polymyxin B which further improves the selectivity, particularly with the inhibition of Pseudomonas spp. Related Supplements : LS0125 B.cepacia Selective Supplement, LS0026 Pseudomonas CFC Selective Supplement
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KM0057 Campylobacter (CCDA) Agar is a selective medium for the isolation of Campylobacter spp., particularly C. jejuni and C. coli, from clinical specimens and foodstuffs. Campylobacter (CCDA) Agar was described by Bolton et al. and formulated to replace blood with a combination of charcoal, ferrous sulphate, and sodium pyruvate. This product is recommended for the isolation of Campylobacter spp. and complies with the requirements of ISO 10272-1:2017, ISO 10272-2:2017, is tested in accordance with ISO 11133:2014, and is a recommended medium in the UK Standards for Microbiology Investigations for isolation of Campylobacter spp. from clinical specimens such as faeces. Bolton et al. recommended incubating inoculated plates at 37°C to improve isolation rates, but incubation at 41.5°C is recommended for the isolation of the commonly encountered thermophilic species (C. jejuni and C. coli). Selectivity is achieved through the addition of E&O LS0010 Campylobacter CCDA Selective Supplement, which includes a broad-spectrum antibiotic, cefoperazone, and amphotericin B to inhibit yeast and fungi.
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This is one of several selective media available for the isolation of Campylobacter spp. in clinical, food and environmental laboratories. Campylobacter agar base is based on the formulation from Bolton and Robertson. The peptone is the source of the required nitrogen, carbon and vitamins. Sodium chloride maintains the osmotic balance in the medium. The medium is enriched with lysed horse blood and made selective by the addition of cefoperazone, to suppress other enteric organisms, and amphotericin to suppress yeast and fungal growth (Preston supplement LS0010). Related Supplements : LS0009 Campylobacter (Skirrow) Selective Supplement, LS0010 Campylobacter (Preston) Selective Supplement, Lysed Blood
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This medium is based on the formulation published by Platt, Atherton & Simpson1 and is used for the cultivation of Taylorella equigenitalis, the causative organism in contagious equine metritis. It is routinely used for culturing swabs taken from the genitalia of mares and stallions. Enzymatic digest of casein and soy peptone supply nitrogen, carbon and vitamins and L-cystine is a required growth factor. Sodium chloride provides osmotic balance and sodium sulphite is present as a reducing agent. The medium is made selective by the addition of CEMO Selective Supplement (LS0041) to control bacteria and fungi from swab samples. References 1. Platt, H., Atherton, J. G. and Simpson, D. J. 1978. Equine Vet J 10, 153–159.
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Cetrimide agar is a selective medium for the isolation and detection of Pseudomonas aeruginosa from pharmaceutical, clinical and cosmetic samples. The formulation is complaint with the requirements of the Harmonised USP/EP/JP. Detection is achieved using the unique ability of P. aeruginosa to produce the water soluble, bright green pigment pyocyanin. The production of this pigment is stimulated by the presence of magnesium chloride and di-potassium sulphate in the medium. The addition of glycerol (10ml/l) is required as this compound serves as an energy source. Cetrimide, a quaternary ammonium compound, is also present to suppress the growth of other Pseudomonas spp. as well as Gram-negative and Gram-positive organisms. Pancreatic digest of gelatin provides the required nitrogen, carbon and vitamins. Related Supplements :
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Charcoal agar is used for the cultivation of fastidious organisms, particularly Bordetella pertussis. Charcoal agar is prepared according to the formulation developed by Mishulow, Sharpe and Cohen. This medium is an efficient substitute for Bordet-Gengou agar in the production of B. pertussis vaccines and can be used as a maintenance medium for stock cultures of Bordetella spp. Beef extract and peptone provide the required nitrogen, carbon and vitamins. Sodium chloride maintains the osmotic balance. Starch and charcoal help in absorbing toxic metabolites that are produced during growth of the organism. Nicotinic acid is an essential growth factor for the growth of Bordetella spp. The addition of cephalexin (LS0018) inhibits accompanying contamination in the samples. NB: This is a basic medium only and contains no additional supplement. If cephalexin is added, it should be noted, that although coliforms are inhibited by this medium, Pseudomonas aeruginosa and some fungi will grow. Related Supplements : LS0018 Bordetella Selective Supplement
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Chromogenic coliform agar (CCA) conforms to the ISO 9308-1 guidelines for the detection, enumeration and isolation of coliforms and more specifically Escherichia coli in water samples by the membrane-filtration technique. The colonial differentiation is provided by the chromogenic substrates, Salmon-GAL and X-glucuronide. Salmon-GAL is used for the detection of ß-D-galactosidase enzymatic activity. X-glucuronide is used for the detection of ß-D-glucoronidase enzymatic activity. β-D-galactosidase, expressed by all coliforms, cleaves the Salmon-GAL substrate and producing red/pink coloured colonies. Unlike other coliforms, Escherichia coli cleaves both Salmon-GAL and X-glucuronide producing a violet/blue coloured colonies. Tryptophan is used to increase detection reliability by improving the indole reaction. The peptones, sodium pyruvate and sorbitol support bacterial growth and simple recovery of sub-lethal thermally injured coliforms. Sodium di-hydrogen phosphate and di-sodium hydrogen phosphate phosphate buffer the medium and sodium chloride is used to achieve osmotic balance. The selectivity is attained by the addition of Tergitol® 7 as it inhibits the growth of Gram-positive bacteria.
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KM0005 CLED Double Indicator Agar (Bevis) is a differential culture medium used clinically for isolation and enumeration of bacteria in urine, from the suspected cases of urinary tract infection (UTI). KM0005 CLED Double Indicator Agar (Bevis) is a differential culture medium which contains cystine, lactose, bromothymol blue, acid fuchsin and is electrolyte deficient to prevent swarming of Proteus species. Bevis changed the original medium by Mackey and Sandys by introducing a second pH indicator to enhance the differentiation of colony characteristics of lactose and non-lactose fermenting organisms. The medium can allow for quantitative determination of urinary pathogens, including Proteus species, when automated systems or calibrated loops are used for inoculation. KM0005 is recommended as a differential primary isolation media by the UK Standards for Microbiology Investigations and tested in accordance with ISO 11133:2014.
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KM0004 CLED Single Indicator Agar is a differential culture medium used clinically for isolation and enumeration of bacteria in urine, from the suspected cases of urinary tract infection (UTI). Originally described by Sandys, CLED (Cystine Lactose Electrolyte Deficient) Single Indicator Agar is a differential medium which contains cystine, lactose, bromothymol blue and is electrolyte deficient to prevent swarming of Proteus species. The medium can allow for quantitative determination of urinary pathogens, including Proteus species, when automated systems or calibrated loops are used for inoculation. KM0004 is recommended as a differential primary isolation media by the UK Standards for Microbiology Investigations and tested in accordance with the principals of ISO 11133:2014.
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Clostridium difficile agar, when supplemented, is used for the isolation of C. difficile from samples. This formulation is a modification of CCFA (Cycloserine-Cefoxitin-Fructose agar) developed by George et al. The proteose peptone act as carbon, nitrogen and vitamin source in this medium. Fructose is a fermentable carbohydrate. Sodium chloride maintains the osmotic balance of the medium. Disodium hydrogen phosphate and potassium dihydrogen phosphate are buffering agents. The magnesium sulfate acts as a source of inorganic ions and the agar is the solidifying agent. This media requires the addition of defibrinated horse blood and Clostridium difficile selective supplement (LS0022).
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KM0001 Columbia Agar is a modification of the original Columbia Agar formulation described by Ellner et al. from Columbia University and is tested in accordance with ISO 11133:2014. Columbia Agar provides a medium that is suitable for use with both defibrinated horse and sheep blood to support the growth and determining haemolytic reactions of a variety of microorganisms. Columbia Agar with the addition of plain or chocolated blood is recommended by the UK Standards for Microbiology Investigations for various purposes, including as a standard, supplementary or primary isolation medium. Columbia agar with blood is a general-purpose, non-selective medium suitable for isolation and cultivation of most organisms including fastidious anaerobes from clinical specimens. Chocolating the blood provides a highly nutritious medium which supports the growth of a wide range of pathogens including the most fastidious organisms and is particularly useful for the cultivation of Haemophilus and Neisseria species. Chocolate agar is the same as blood agar except that during preparation the red blood cells are lysed when added to molten agar base, which gives the medium a chocolate-brown colouration and from which the agar gets its name. As a result, cell lysis releases intracellular nutrients such as haemoglobin, haemin (X factor), and the coenzyme nicotinamide adenine dinucleotide (NAD or V factor) into the agar. These growth factors are available to fastidious organisms during growth. Related Supplements : Defibrinated Horse Blood, Defibrinated Sheep Blood, Horse Blood Lysed, LS0008 Staph/Strep Selective Supplement, LS0017 Neomycin Selective Supplement, LS0022 Clostridium difficle Selective Supplement, LS0012 Bacitracin Selective Supplement, LS0011 Campylobacter Growth Supplement, LS0009 Campylobacter (Skirrow) Selective Supplement
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This medium is used for the cultivation and enumeration of Lactobacillus spp. MRS agar is based on the formulation by de Man, Rogosa and Sharpe. The peptone, yeast extract and beef extract provides the reuired carbon, nitrogen and vitamin source. Glucose is a fermentable carbohydrate. The magnesium sulphate and manganese sulphate act as growth stimulants. Potassium phosphate is a buffering agent. Selectivity of the medium is achieved through the use of ammonium citrate and sodium acetate, inhibiting microorganisms such as streptococci and moulds. The addition of the surfactant Tween 80 is required to facilitate the uptake of nutrients from Lactobacillus spp.
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This medium is intended for the cultivation and enumeration (via the Miles and Misra technique) of Lactobacillus spp. from a variety of sources and can be used in conjunction with MRS Agar (KM0080). This medium is a modification on the formulation developed by de Man, Rogosa and Sharpe.(1) The peptones, beef extract, yeast extract provides the required carbon, nitrogen and vitamins in this medium. Glucose is a fermentable carbohydrate. Potassium phosphate is a buffering agent. Selectivity of the medium is achieved through the use of ammonium citrate and sodium acetate, inhibiting microorganisms such as streptococci and moulds. The magnesium sulphate, manganese sulphate and polysorbate 80 act as growth stimulants. The medium can be made more specific for lactobacilli generally by lowering the pH to between 5.0 and 5.5. This has the effect of inhibiting most streptococci that may otherwise grow on the medium and can be readily confused with lactobacilli.
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This is a selective medium for the isolation of Salmonella spp. and Shigella spp. from clinical specimens and environmental samples. This formulation was developed by Hynes through a modification of Leifson’s DCA medium. The peptone provides the required carbon, nitrogen and vitamins. Lactose is a fermentable carbohydrate and neutral red is a pH indicator. Sodium thiosulphate and ferric citrate are used to detect hydrogen sulphide production indicated by the black centred colonies of hydrogen sulphite positive organisms. Sodium desoxycholate and sodium citrate inhibit most Gram-positive organisms.
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A selective medium for the isolation and detection of dermatophytic fungi originally developed by Taplin et al. Dermatophytes appear as fluffy colonies with reddening of the medium while other fungi cause the medium to become yellow due to acid production. Yeasts will also grow on this medium but are readily distinguished by their distinct white/creamy colonies and distinctive smell. Soy peptone provides the required carbon, nitrogen and vitamins. Glucose is a fermentable carbohydrate. The inclusion of phenol red assists in the differentiation between saprophytic and environmental fungi. Although the low pH (pH 5.5) of the medium inhibits most bacteria chloramphenicol and cycloheximide are often added to further reduce the risk when processing material that may be more heavily contaminated. NB: Prolonged incubation should be avoided as this may cause fungi other than dermatophytes to turn the medium red. Related Supplements : LS0050 Chloramphenicol Selective Supplement
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Dichloran Rose Bengal Chloramphenicol (DRBC) agar is based on the formulation described by King et al. and is used for the selective isolation and enumeration of yeasts and moulds from food samples. The peptone provides the required carbon, nitrogen and vitamins. Glucose is a fermentable carbohydrate. Potassium di-hydrogen phosphate is a buffering agent and magnesium sulfate is a source of divalent ions and sulfate. In order to curtail the size of the colony diameters of spreading fungi, the antifungal agent dichloran is added to the base and the pH is reduced to 5.6. Rose Bengal suppresses growth of bacteria and restricts the size and height of colonies of more rapidly growing molds. The inclusion of chloramphenicol ensures the suppression of bacteria present in environmental and food samples. Rose Bengal is absorbed by yeast and mold colonies and this further aids in their enumeration. Occasionally reduced recovery of yeasts may be encountered due to the increased activity of Rose Bengal at pH 5.6.
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DNase agar is used primarily in clinical laboratories to differentiate pathogenic Staphylococcus aureus from other staphylococci based on deoxyribonuclease (DNase) activity. The tryptone provides the required carbon, nitrogen and vitamins. Sodium chloride maintains the osmotic balance. The addition of DNA to the base medium provides a simple method to check for DNase activity. Following incubation of the inoculated medium, the surface of the medium is flooded with a small quantity of 1M hydrochloric acid to precipitate the DNA. This results in the medium turning opaque. Organisms that can produce sufficient quantity of a DNase enzyme will hydrolyse the DNA resulting in a clear area around the colonies. Whereas DNase negative organisms will not produce clearing. NB: As with most tests of this type a positive result should not be taken in isolation and other appropriate tests, e.g. coagulase test, latex agglutination etc., should be carried out.
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DNase agar is used primarily in clinical laboratories to differentiate pathogenic Staphylococcus aureus from other staphylococci based on deoxyribonuclease (DNase) activity. The tryptone provides the required carbon, nitrogen and vitamins. Sodium chloride maintains the osmotic balance. The addition of DNA to the base medium provides a simple method to check for DNase activity. The methyl green fades into a colourless compound if the DNA in the medium is depolymerised. Organisms that can produce sufficient quantity of a DNase enzyme will hydrolyse the DNA resulting in a clear area around the colonies. Whereas DNase negative organisms will not produce clearing. NB: As with most tests of this type a positive result should not be taken in isolation and other appropriate tests, e.g. coagulase test, latex agglutination etc., should be carried out.
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Edwardsiella ictaluri medium is a selective medium for the isolation of Edwardsiella spp. based on the formulation by Shotts et al. (1) Edwardsiella spp. may be differentiated from other microorganisms on this medium due to its colony morphology. The peptones provide the required carbon, nitrogen and vitamins. Sodium chloride maintains the osmotic balance. Colistin and bile salts are selective agents to inhibit most Gram-negative and Gram- positive organisms. Mannitol is a fermentable carbohydrate and bromthymol blue is a pH indicator. Phenylalanine and ferric citrate are used to detect phenylalanine deaminase activity in Proteus spp. Agar is a solidifying agent. Shotts et al. (1) noted Proteus spp. swarming can overgrow on a mixed cultured sample. Therefore, a selective supplement (LS0021) may be added to the medium to restrict Proteus spp. swarming
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E.E. Broth is recommended as an enrichment medium when examining food and feedstuffs for Enterobacteriaceae. It is a modification of Brilliant Green Bile Broth, with an improved buffering capacity to encourage early growth and prevent autosterilization. E.E. Broth uses glucose instead of lactose to make the medium a test for all enterobacteria including non-lactose fermenting organisms. This formulation complies with the Harmonized USP/EP/JP. Nitrogen is supplied by the gelatin peptone whilst glucose serves as the fermentable carbohydrate source. Oxbile and brilliant green are the selective agents helping to suppress Gram-positive non- target organisms. Auto sterilisation is prevented through the buffer system composed of potassium dihydrogen phosphate and disodium hydrogen phosphate.
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Eosin Methylene Blue agar (EMB) is a selective medium primarily for the isolation of coliforms from clinical, food and environmental samples. This is the modified formulation of EMB proposed by Levine with a higher concentration of lactose and the sucrose omitted. The peptone provides the required carbon, nitrogen and vitamins. Lactose is a fermentable carbohydrate and di-potassium phosphate is a buffer. Eosin Y and methylene blue are indicators. Methylene blue is also a selective agent. During strong acidic conditions, the dyes impart a metallic sheen to certain lactose fermenters, such as Escherichia coli.
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KM0006 Fastidious Anaerobe Agar is a general-purpose medium, when supplemented with blood and any necessary selective agents, can be used for the isolation and culture of fastidious anaerobes from clinical specimens. Anaerobic infections are common infections caused by anaerobic bacteria. These bacteria occur naturally and are the most common flora in the body; in their natural state, they do not cause infection. Anaerobic bacteria can infect deep wounds, tissue, and internal organs where there is little oxygen after injury or surgery. Infection by anaerobic bacteria is normally characterized by formation of abscesses, foul smelling discharge, pus, and free gas in tissue, leading to tissue degradation. KM0006 is recommended as a standard, primary isolation or supplementary medium by the UK Standards for Microbiology Investigations and tested in accordance with the principles of ISO 11133:2014. Related Supplements : LS0015 Actinomycete Selective Supplement, LS0017 Neomycin Selective Supplement, LS0022 Clostridium difficile Selective Supplement, LS0023 Clostridium perfringens Selective Supplement, BM0140 Egg Yolk Emulsion
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This medium is for the growth of fastidious anaerobes, particularly Bacteroides spp. Fastidious anaerobe broth is also suitable for anaerobic blood culture. The peptone and yeast extract provides the required carbon, nitrogen and vitamins. Haemin, vitamin k and L-cysteine HCl are growth factors required by some anaerobes. Sodium thioglycollate and L-cysteine HCl reduce the Eh of the medium and the agar helps maintain the Eh. Resazurin is a redox indicator and sodium chloride maintains the osmotic balance. NB: For best results it is recommended that the medium be heated in a boiling water bath, with the cap loosened, and then allowed to cool, with the cap tightened, immediately before use. The cap must be replaced on the container immediately after inoculation.
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Half-Fraser broth is a selective enrichment broth for the identification and isolation of Listeria spp., primarily from food and environmental samples. The antibiotics, nalidixic acid and acriflavine, are already included in the formulation so only ferric ammonium citrate (LS5004) need be added to make the complete Half-Fraser broth. The peptones provide carbon, nitrogen and vitamins, sodium chloride provides osmotic balance and the phosphate buffer system maintains pH. Lithium chloride inhibits enterococci and the antibiotics make the medium highly selective. Listeria spp. hydrolyse aesculin to aesculetin which forms a confirmatory dark brown or black complex with Fe3+ ions.
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KM0016 GC Agar is based on the formulation described by Thayer and Martin that was modified from the formulation development by Johnson for the isolation of gonococcus cultures and is recommended primarily for the isolation of Neisseria gonorrhoeae and Neisseria meningitidis; however, it is also capable of supporting the growth of most fastidious micro-organisms from clinical specimens, and is recommended by the UK Standards for Microbiology Investigations for a variety of clinical samples, and falls under the category of selective media for pathogenic Neisseria spp. according to the Clinical and Laboratory Standards Institute. Enrichment of KM0016 GC Agar is usually attained using lysed blood but haemoglobin powder or chocolated blood are suitable alternatives. Further enrichment may be provided by the addition of Suplex Supplement (E&O BM0478) which contains yeast extract and glucose. Selective plated media variants of KM0016 GC Agar, may be prepared by the addition of various selective supplements such as LCAT Selective Supplement (E&O LS0001) or VCAT Selective Supplement (E&O LS0002). These supplements suppress most microbiota present in clinical specimens. Related Supplements : BM0478 Suplex, LS0001 GC LCAT Selective Supplement, LS0002 GC VCAT Selective Supplement
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KM0215 Half Fraser Broth Complete is used to make Half Fraser Broth enrichment media for the isolation of Listeria species from food and environmental samples.
Developed in 1988 by Fraser (1), the formulation can be made as primary enrichment Half Fraser Broth and secondary enrichment Fraser Broth according to the requirements of ISO 11290-1:2017 (2). KM0215 is recommended by the International Standardization Organization (2) and tested in accordance with the principals of ISO 11133:2014 (3).
The nalidixic acid, acriflavine, and ferric ammonium citrate are already included in the formulation, so no additional supplements are required to make complete Half Fraser Broth.
- Fraser, J. and Sperber, W. (1988) Rapid Detection of Listeria spp. in Food and Environmental Samples by Esculin Hydrolysis. Journal of Food Protection, 51(10), pp.762-765. 2. International Standardization Organization (2017) 11290-1:2017 Microbiology of the food chain - Horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp. - Part 1: Detection method. Geneva: ISO. 3. International Standardization Organization (2014) 11133:2014 Microbiology of food, animal feed and water — Preparation, production, storage and performance testing of culture media. Geneva: ISO.
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Hektoen enteric agar was developed by King and Metzger as a differential selective medium for the isolation of Shigella spp. and Salmonella spp. species from enteric pathological samples. The meat peptone and yeast extract provide the required nitrogen, carbon and vitamins. Lactose, sucrose and salicin are fermentable carbohydrates. Bromothymol blue is added as a pH indicator in order to identify carbohydrate fermenting organisms. The combination of ferric ammonium citrate and sodium thiosulfate allows the production of hydrogen sulphide. Hydrogen sulphide positive colonies produce black centred colonies. Sodium chloride maintains the osmotic balance. The bile salts and acid fuchsin inhibit Gram-positive organisms.
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This is a selective medium for the isolation of Helicobacter pylori from clinical samples. The medium is based on a modification of Campylobacter CCDA Blood Free Medium with charcoal, ferrous sulphate and sodium pyruvate replacing the horse blood. The medium is made selective by the addition of vancomycin and cefsulodin, to suppress other bacteria, and amphoteracin to inhibit yeasts. Horse serum (10%) is also added to promote optimum growth of Helicobacter spp. Related Supplements : LS0031 Helicobacter pylori Selective Supplement, SHS500 Sterile Horse Serum 500ml
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Hoyles agar is selective culture medium for the isolation and differentiation of Corynebacterium diphtheriae types. Hoyles agar allows for rapid growth of the organisms and normally 18 hours incubation should be sufficient for a diagnosis. As the medium is highly selective, inoculation should be by rubbing the swab (or other material) over the entire surface of the agar, there is no need to spread the inoculum with a loop indeed doing so can cause the organism to be missed especially when they are present only in small numbers. The Elek method can be used to determine the toxigenicity of any C. diptheriae strains. The beef extract and peptone act as a source of nitrogen, carbon and vitamins. The sodium chloride maintains osmotic balance. This medium should be used in conjunction with two supplements: lysed horse blood at 50 ml/l and 10ml/l of potassium tellurite 3.5% solution (BM2230). Lysed horse blood provides added nutrients to the medium and potassium tellurite inhibits Gram-negative and several Gram-positive microorganisms. Related Supplements : BM2290 3.5% Potassium Tellurite Solution, Horse Blood Lysed
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Kligler iron agar is used to differentiate between some of the enterobacteriacae on the basis of three reactions: fermentation of lactose and glucose and the production of hydrogen sulphide. Kligler iron agar is a modification of the original formulation developed by Kligler. It incorporates the principles of Russell’s double sugar agar with Kligler ‘s lead acetate agar. The peptone provides the required nitrogen, carbon and vitamins. Lactose and glucose are carbohydrates. Acid production from their fermentation is detected by the phenol red pH indictor. Sodium thiosulphate is reduced to hydrogen sulphide which is detected by the ferric citrate indicator. Sodium chloride maintains the osmotic balance.
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Lactalbumin hydrolysate is obtained through enzymatic hydrolysis of lactalbumin protein in milk whey. This product is rich in essential amino acids as well as providing a source of nitrogen, carbon and vitamins. It is commonly utilized in media for tissue culture and for the production of vaccines as well as being useful for bacterial and fermentation media.
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Lactose broth is used for the performance and confirmation of the Presumptive Test for coliforms in water and dairy samples. This medium is also frequently used as a pre-enrichment medium when testing foods, water samples and dairy products for Salmonella spp. Beef extract and gelatin peptone provide the required carbon, nitrogen and vitamins in this medium. Lactose is a fermentable carbohydrate. Fermentation of lactose is detected by the production of gas
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Lauryl Tryptose Broth is a selective medium for the detection of coliforms in water and wastewater. This formulation is based on the Mallmann and Darby formulation. (1) The tryptone provides the required carbon, nitrogen and vitamins in this medium. Lactose is a fermentable carbohydrate. The sodium chloride maintains the osmotic balance whilst the dipotassium hydrogen phosphate and potassium hydrogen phosphate act as buffers. Sodium lauryl sulphate is a selective again used to inhibit non-coliforms. References (1) Mallmann, W. L. and Darby, C. W. 1941. Uses of a lauryl sulphate tryptose broth for the detection of coliform organisms. Am J. Public Health. 31:127
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LB agar (Lennox) is used for the cultivation and maintenance of recombinant strains of Escherichia coli and is based on the formulation stated by Lennox. This medium is typically used in molecular biology procedures such as the detection of phage or plasmid transformed bacteria. LB agar (Lennox) is prepared using 5 g/L of sodium chloride and this level varies from that described by Miller. This allows for additional sodium chloride to be added at the point of preparation if required. Enzymatic digest of casein provides the required nitrogen and carbon for organism growth. Vitamin B complex required by recombinant strains of E. coli are supplied by the yeast extract. Sodium chloride maintains the osmotic balance and provides sodium ions for membrane transport. Related Supplements : LS0035 Ampicillin Selective Supplement, LS0037 Kanamycin Selective Supplement