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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 -
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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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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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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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
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Luria Bertani (LB) agar (Miller) is used for the cultivation and maintenance of recombinant strains of Escherichia coli and is based on the formulation stated by Miller. This medium is typically used in molecular biology procedures such as the detection of phage or plasmid transformed bacteria. LB agar (Miller) is prepared using 10 g/L of sodium chloride and this level varies from that described by Lennox. 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
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KM0015 Legionella Agar Base is the base for media used in the isolation of Legionella species from clinical, water and environmental samples and is tested in accordance with ISO 11133:2014. Legionella Agar, initially known as F-G agar, was modified by Feely et al. by replacing starch with charcoal, and casein hydrolysate with yeast extract which resulted in better recovery of Legionella pneumophila. The medium requires supplementation with ferric pyrophosphate as a source of iron, L-cysteine, an essential amino acid for the growth of Legionella spp. and α-ketoglutaric acid, which acts as a growth stimulant. ACES buffer/potassium hydroxide is also added to maintain the optimal pH of 6.9 for growth of Legionella spp. Omission of the L-cysteine produces a confirmation medium that can be used to test presumptive Legionella spp. as isolates will not be able to grow. Many variants of Legionella Agar can be created from KM0015 Legionella Agar Base by the addition of various supplements (GVPC is the most popular for water testing and BMPA for clinical testing).
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Letheen agar modified is intended for use in the isolation of microorganisms from cosmetics. The casein peptone, meat peptone, beef extract and yeast extract act as carbon, nitrogen and vitamin sources in this medium. Glucose is a fermentable carbohydrate. Sodium chloride maintains the osmotic balance of the medium. The sodium bisulfite, lecithin and polysorbate 80 inactivates quaternary ammonium compounds. Polysorbate 80 neutralises phenols, formalin, hexachlorophene, and in combination with the lecithin, ethanol.
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Listeria isolation medium (Oxford) is based on the formulation described by Curtis et al. and is used for the isolation and identification of Listeria spp. in food and clinical laboratories. Columbia agar base provides the required carbon, nitrogen and vitamins in the medium. Lithium chloride is included to inhibit enterococci. Aesculin is present as an indicator; Listeria spp. will hydrolyse aesculin and the associated reaction with the ferric ammonium citrate gives rise to a black precipitate around the colonies. Selectivity is enhanced by addition of Listeria Oxford selective supplement (LS0030). This contains acriflavine, cefoxitin, colistin, fosfomycin and amphotericin to inhibit any yeasts present and some other Gram-positive and Gram-negative organisms that may be present in specimens. Related Supplements : LS0030 Listeria Oxford Selective Supplement
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Long and Hammer agar is for the detection of aerobic microorganisms in fish and fish products. This media is recommended for the determination of spoilage organisms in fresh and lightly preserved seafoods. (1) The proteose peptone acts as a nitrogen, carbon and vitamin source. Gelatine is a protein source and solidifying agent. Sodium chloride maintains the osmotic balance and di-potassium hydrogen phosphate is a buffer. 1) Nordic Committee on Food Analysis. (2006) Aerobic count and specific spoilage organisms in fish and fish products. NMKL Method No 184
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Lowenstein-Jensen medium is based on the original formulation of Lowenstein that was subsequently modified by Jensen. It is used with fresh egg and glycerol for the isolation and presumptive identification of Mycobacterium spp., particularly Mycobacterium tuberculosis, from clinical samples. L-Asparagine and Potato starch are sources of nitrogen and vitamins in the medium. Potassium hydrogen phosphate and magnesium sulphate enhance organism growth and act as buffers. Sodium citrate and malachite green are selective agents that have inhibitory effects on organisms other than the mycobacteria. Malachite green is also incorporated into the medium to provide a colour contrast between the colonies and the medium. The required addition of egg emulsion provides the fatty acids and protein required for the metabolism of mycobacteria. Glycerol is also added which is said to enhance the growth of Mycobacterium tuberculosis however other strains of mycobacteria, particularly Mycobacterium bovis, may be inhibited by its presence. Subsequently, sodium pyruvate (12.5 g/600ml) may be used as an alternative to glycerol to encourage the growth of Mycobacterium bovis.
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MacConkey agar No. 3 is a selective medium primarily for the isolation of Enterobacteriacae from waters and sewage. This media differs from the original MacConkey formulation in that as well as bile salts, crystal violet has been included as an additional selective agent. This has the effect of inhibiting Gram-positive organisms. The peptone acts as a nitrogen, carbon and vitamin source. Sodium chloride maintains osmotic balance. Lactose is a carbohydrate and during its fermentation causes a confined pH drop around the bacterial colony. This causes a colour change in the pH indicator, neutral red and bile precipitation.
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KM0124 MacConkey Agar with Salt is a selective medium for the isolation and differentiation of bile tolerant Gram-negative (enteric) and Gram-positive (staphylococci and enterococci) organisms in all areas of bacteriology. MacConkey Agar with Salt is based on the formulation by MacConkey in 1900. Staphylococcus and Enterococcus spp. are able to grow due to the omission of crystal violet from this formulation. This medium is recommended by the World Health Organisation and the Department of Health for the bacteriological examination of water, and KM0124 is recommended as a differential primary isolation media by the UK Standards for Microbiology Investigations.
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MacConkey Agar without Salt and Crystal Violet, based on the formulation by Rappaport and Henig, is a differential medium for the isolation and differentiation of lactose and non-lactose fermenting enteric bacteria while also restricting the swarming of Proteus species. KM0011 is recommended as a differential primary isolation media by the UK Standards for Microbiology Investigations, The Microbiology of Drinking Water and tested in accordance with ISO 11133:2014. Related Supplements : LS0189 Cefotaxime Supplement (1mg/L)
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Malt extract agar is used for the detection, isolation and enumeration yeasts and moulds. The high carbohydrate content of the medium ensures rapid growth of yeasts and moulds. Malt extract and mycological peptone provide the carbon, protein and nutrient sources required by the organisms. The acidic nature of this medium allows for optimal growth of moulds and yeasts while restricting bacterial growth. Malt extract agar can also be used for the cultivation of fungi, although with the prolonged incubation necessary, cultures may become overgrown by bacteria. The selectivity of the medium can be increased by lowering the pH to 3.5-4.0 or by the addition of selective agents such as chloramphenicol. Related Supplements : LS0050 Chloramphenicol Selective Supplement
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KM0031 Mannitol Salt Agar is a selective medium for the isolation of Staphylococcus aureus from clinical samples, food, cosmetics and water samples. Chapman showed that adding a higher level of sodium chloride to Mannitol Salt Agar allowed for the recovery of pathogenic staphylococci and inhibits most organisms. Coagulase positive staphylococci (e.g., S. aureus) produce yellow colonies and a surrounding yellow medium while coagulase negative staphylococci produce red colonies and no colour change of the phenol red indicator. The medium conforms to the requirements of the Harmonised EP/USP/JP, is recommended as a primary isolation media by the UK Standards for Microbiology Investigations and tested in accordance with ISO 11133:2014 and Clinical and Laboratory Standards Institute. This medium is also included in the Bacteriological Analytical Manual for cosmetics testing.
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Marine agar is a medium used to support the growth of numerous marine bacteria. Originally formulation by ZoBell (1), its high saline levels are to simulate a close approximation of seawater which allows the marine bacteria to grow (2). The sodium chloride and marine minerals provide the high salinity needed for marine bacteria to thrive and aims to simulate sea water. The peptones provide the necessary nitrogen, amino acids and vitamins for growth. Agar is the solidifying agent. References (1) ZoBell, C.E. 1941. Studies on Marine Bacteria. I. The cultural requirements of heterotrophic aerobes. J. Mar. Res. 4, 42-75. (2) Lyman, J. and Fleming, R. H. 1940. Composition of Seawater. J. Mar. Res. 3, 134-146
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Membrane Lauryl Sulphate Agar is used for the enumeration of Escherichia coli and other coliforms found in filter membranes used in water sample testing. The broth base, originally named Membrane Enriched Teepol broth,(1) was updated when Teepol 610 was removed from the formulation and replaced by sodium lauryl sulphate.(2&3) The peptones, yeast extract and lactose act as carbon, nitrogen and vitamin sources in this medium. The phenol red is added as a pH indicator to detect the fermentation of lactose to differentiate the coliforms. Sodium lauryl sulphate is an inhibitory agent. References (1) Burman, N. P., 1967. Development of membrane filter techniques. II. Adaptation to routine and special requirements. Proc. Soc. Wat. Treat. Exam., 16:40 (2) Joint Committee of PHLS and the Standing Committee of Analysis. 1980. J. Hyg. Camb., 85:181 (3) Stanfield, G. and Irving, T. E., 1981. A suitable replacement for Teepol 610 in the selective isolation of coliforms from marine waters and sewage. Water Research. 15:469-474
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KM0183 Mueller Hinton Agar is used as a base to make various media for antimicrobial susceptibility testing by the disk diffusion method as described by Bauer-Kirby. First described by Mueller and Hinton, Mueller Hinton Agar has been approved as the definitive medium for antimicrobial susceptibility testing by European Committee on Antimicrobial Susceptibility Testing (EUCAST), Clinical and Laboratory Standards Institute (CLSI) and is compliant with ISO16782:2016. KM0183, when prepared as unsupplemented Mueller Hinton Agar, is suggested by EUCAST and CLSI for non-fastidious organisms such as Enterobacterales, Pseudomonas spp., Stenotrophomonas maltophilia, Acinetobacter spp., Staphylococcus spp., Enterococcus spp., Aeromonas spp., Achromobacter xylosoxidans, Vibrio spp., Bacillus spp. and Burkholderia pseudomallei. KM0183, when prepared supplemented with 5% horse blood and nicotinamide adenine dinucleotide (NAD), is suggested by EUCAST and CLSI for fastidious organisms such as Streptococcus pneumoniae, Streptococcus groups A, B, C and G, Viridans group streptococci, Haemophilus influenzae, Moraxella catarrhalis, Listeria monocytogenes, Pasteurella multocida, Campylobacter jejuni and coli, Corynebacterium spp., Aerococcus sanguinicola and urinae and Kingella kingae. Additionally, according to the UK Standards for Microbiology Investigations, when supplemented appropriately, Mueller Hinton Agar can be used in the process of identifying Haemophilus species and the HACEK group of organisms and Helicobacter species from clinical specimens. Related Supplements : LS0005 NAD Supplement (20mgs/L), Defibrinated Horse Blood
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Mycological agar is a selective medium for the isolation of pathogenic fungi, particularly dermatophytes, from clinical specimens. Enzymatic digest of soybean meal provides the required carbon, nitrogen and vitamins. Glucose is an energy source for the metabolism of fungi. The addition of chloramphenicol further reduces the risk of bacterial contamination when processing material that may be heavily contaminated with coliforms. Cycloheximide should also be added (0.4g/L) to suppress the growth of commensal yeasts and saprophytic fungi.
