Selective enrichment of following microorganism Media Positive test organism Page

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1 Selective enrichment of following microorganism Media Positive test organism Page Escherichia coli m-total Count Media Escherichia coli ATCC MI-Broth Escherichia coli ATCC Tryptic Soy Broth Escherichia coli ATCC M -Endo Coliform Broth Escherichia coli ATCC Total Count Media with TTC Escherichia coli ATCC Coliform MacConkey with MUG Escherichia coli ATCC M-FC/ M-FC Rosolic Acid Escherichia coli ATCC Fecal Streptococci KF-Streptococcus Broth Streptococcus faecalis ATCC Pseudomonas aeruginosa Cetrimide Broth Pseudomonas aeruginosa ATCC in purified water Pseudomonas Broth Pseudomonas aeruginosa ATCC Staphylococci Mannitol Salt Broth Staphylococcus aureus ATCC Enterococci Enterococcus Broth Streptococcus faecalis ATCC Soft drinks Selective enrichment of following microorganism Media Positive test organism Page Escherichia coli Wallerstein Differential Broth Escherichia coli ATCC Coliform M -Endo Coliform Broth Escherichia coli ATCC Yeast and Mold M-Green Select Broth Saccharomyces cerevisiae ATCC M-Green Yeast and Mold Saccharomyces cerevisiae ATCC Lactobacillus acidophilus Orange Serum Broth Lactobacillus acidophilus ATCC Beer and wine Selective enrichment of following microorganism Media Positive test organism Page Escherichia coli Wallerstein Differential Broth Escherichia coli ATCC Coliform M -Endo Coliform Broth Escherichia coli ATCC Yeast and Mold M-Green Select Broth Saccharomyces cerevisiae ATCC Wallerstein Nutrient Broth Saccharomyces cerevisiae ATCC

2 Products Media Quick Media Selection Guide Quick Media Selection Guide Dairy products Selective enrichment of following microorganism Media Positive test organism Page Escherichia coli Wallerstein Differential Broth Escherichia coli ATCC Coliform MacConkey with MUG Escherichia coli ATCC Yeast and Mold Potato Dextrose Broth Saccharomyces cerevisiae ATCC Lactobacillus MRS Broth Lactobacillus plantarum ATCC Food Selective enrichment of following microorganism Media Positive test organism Page Escherichia coli Tryptic Soy Broth Escherichia coli ATCC MIBlue Escherichia coli ATCC Coliform MacConkey with MUG Escherichia coli ATCC Fecal Streptococci KF-Streptococcus Broth Streptococcus faecalis ATCC Yeast and Mold Potato Dextrose Broth Saccharomyces cerevisiae ATCC Lactobacillus MRS Broth Lactobacillus plantarum ATCC Pharmaceutical and cosmetics Selective enrichment of following microorganism Media Positive test organism Page Escherichia coli Tryptic Soy Broth Escherichia coli ATCC M-Endo Coliform Broth Escherichia coli ATCC Coliform MacConkey with MUG Escherichia coli ATCC Fecal Streptococci KF-Streptococcus Broth Streptococcus faecalis ATCC Yeast and Mold Sabouraud Dextrose Broth Saccharomyces cerevisiae ATCC Staphylococci Mannitol Salt Broth Staphylococcus aureus ATCC Pseudomonas aeruginosa Cetrimide Broth Pseudomonas aeruginosa ATCC Pseudomonas Broth Pseudomonas aeruginosa ATCC

3 Media Brilliant Green Bile Broth 2 % Products Characteristics E. coli ATCC Growth / gas E. aerogenes ATCC Growth / gas E. faecalis ATCC Inhibited S. aureus ATCC inhibited Additional information: To prevent the growth of accompanying microorganisms in this media an increased concentration of brilliant green should be added. Salmonella, for example, are not able to ferment either lactose or sucrose. For this reason the lactose contained in this medium allows identification of accompanying, weakly lactose-positive or lactose-negative organisms. Order information Brilliant Green Bile Media Bottled broth 9 ml vial, Durham tube

4 Products Media Cetrimide Broth Cetrimide Broth Used for the isolation and determination of Pseudomonas aeruginosa. Cetrimide Broth complies with the recommendations of the United States Pharmacopeia and also European Pharmacopeia. The formulation of this medium corresponds to the specifications in the DIN Norm Pseudomonas aeruginosa is characterized by the production of pyocyanin (a blue green, water soluble, nonflourescent, phenazine pigment), which is stimulated by the inclusion of magnesium chloride and potassium sulfate in the broth. Cetrimide (N-cetyl-NNN-trimethylammonium bromide) is added to inhibit bacteria other than Pseudomonas aeruginosa. Its action as a quaternary ammonium cationic detergent causes nitrogen and phosphorus to be released from bacterial cells other than Pseudomonas aeruginosa. Pseudomonas aeruginosa ATCC 10145, incubated hours at 35º C hours at 35º C. Cetrimide Broth: Pure Culture of Pseudomonas aeruginosa ATCC Per liter of Water adjusted to ph 7.2 ± 0.2 Peptone Magnesium chloride Potassium sulfate Cetrimide Glycerol 20.0 g 1.4 g 0.3 g 10.0 ml Pyocyanin blue-green pigmentation surrounding growth is positive for Pseudomonas aeruginosa. No color development is negative for Pseudomonas aeruginosa. Characteristics Coloring P. aeruginosa ATCC Growth Blue/green E. coli ATCC inhibited S. aureus ATCC inhibited Historical background: Harper and Canton followed by Hood described the use of cetrimide (cetyl-trimethylammonium bromide) for selective isolation of Pseudomonas aeruginosa. Lawbury used cetrimide in a 0.1 % concentration for clinical application. Sawbury and Collins later reported a modification of the concentration of cetrimide required for selectivity. The introduction of Cetevlon (cetrimide) stimulated a new study to determine the minimum concentration of cetrimide required for selective isolation of Pseudomonas aeruginosa for mixed clinical flora. The new experiments demonstrated that a concentration of 0.03 % using the much improved Cetavlon was sufficient for selectivity. Brown and Sawbury introduced the use of a new improved cetrimide agar in By combining the Medium B of King, Ward and Raney with the 0.03 % cetrimide concentration previously introduced, they developed a medium that would support the grow of most desired organisms. Order information Cetrimide Broth Ampouled Media 2 ml Bottled broth 50 ml * 39

5 Media EC Broth Products Growth at 44.5º C E. coli ATCC Growth / gas E. aerogenes ATCC Growth / no gas E. faecalis ATCC inhibited Growth at 37º C E. coli ATCC Growth / gas E. aerogenes ATCC Growth / gas E. faecalis ATCC inhibited Historical background: EC Broth was developed by Hajna and Perry for use in the detection of coliform bacteria at 37 C and Escherichia coli at 44.5 C. This buffered lactose broth was designed to improve the methods of detection of contamination of waters, milk and shellfish. Bile salts were incorporated to inhibit the growth of gram-positive cocci and sporeformers which frequently were responsible for false-positive results obtained when using lactose broth or lauryl tryptose broth. The EC Broth formula conforms to that recommended by the APHA for use in determining the source of water pollution. Through employment of the elevated temperature confirmatory test procedure, differentiation can be made between coliforms of fecal origin (intestine of warm-blooded animals) and coliforms from other sources Order information EC Media Bottled broth 9 ml, vials, Durham tubes

6 Products Media EC Broth with MUG EC Broth with MUG Used for the detection of Escherichia coli in water and food samples by a fluorogenic procedure. The presence of Escherichia coli is detected by the appearance of fluorescence throughout the tube. incubated 24 hours at 35 37º C Enterobacter aerogenes ATCC 13048, incubated 24 hours at 35 37º C. EC-Broth: Vial left: Control; Vial right: Broth inoculated with Escherichia coli ATCC to ph 6.9 ± 0.2 The presence of the fluorescence using a long-wave UV light source confirms the presence of Escherichia coli, and any further confirmation is not required. MUG detects anaerogenic strains, which may not be detected in the conventional procedure. Lactose is a source of energy. Casein peptone provides additional nutrients. The mixture of bile salts is inhibiting for gram-positive bacteria, particularly bacilli and fecal streptococci. The substrate 4-methylumbelliferyl-β- D-glucuronide is hydrolyzed by an enzyme, β-glucuronidase, possessed by most Escherichia coli and a few strains of Salmonella, Shigella and Yersinia, to produce a fluorescent end product, 4-methylumbelliferone. Pancreatic Digest of Casein Lactose Bile Salts Mixture Dipotassium Phosphate Monopotassium Phosphate Sodium Chloride 4-Methylumbelliferylβ-D-glucuronide 20.0 g 1.5 g 4.0 g 1.5 g 50 mg Growth at 44,5 C E. coli ATCC Growth/gas/ fluorescence E. aerogenes ATCC inhibited fluorescence E. faecalis ATCC inhibited Growth at 35 C E. coli ATCC Growth/gas/ fluorescence E. aerogenes ATCC Growth/gas/ no fluorescence E. faecalis ATCC inhibited Order information EC Media with MUG Bottled broth 9 ml, vial Bottled broth 9 ml vial, no Durham Tubes

7 Media Enterococcus Broth Products Enterococcus Broth Selected media for use in membrane filtration procedures for the isolation and enumeration of enterococci in food, water and other materials. Enterococcus broth is a modified version of the improved media described by Slanetz and Bartley with TTC. The membrane filtration method is simple to perform, does not require confirmation and permits a direct count of enterococci in 48 hours. Enterococci appear as pink to dark maroon colonies from mm in diameter. Organism Growth/Coloring E. faecalis ATCC Pink to red colonies E. coli ATCC Inhibited Positive control. Enterococcus faecalis ATCC 19433, incubated at 35º C for 24 hours. Escherichia coli ATCC incubated at 35º C for hours. Sterility test: 14 days plated sterility test. Enterococcus Broth: A pure culture of Enterococcus faecalis ATCC appears pink to red on this media. to ph 7.2 ± 0.2 Yeast extract Casein Dextrose Papaic digest of soybean meal Potassium phosphate Sodium azide Triphenyltetrazoliumchloride1% g 4.0 g 0.4 g 10 ml Additional informations: The presence of sodium azide function as inhibitor for the growth of the entire accompanying Gram-negative microbial flora. As described above Enterococci reduce TTC and therefore, their colonies appear pink to dark maroon in color. Furthermore an improved selectivity for enterococci can be obtained by adding additives like carbonate and Tween 80 to the media (Lachica and Hartman,1968). Order information Enterococcus Media Ampouled Media 2 ml

8 Products Media Eugon Broth Eugon Broth Used for the cultivation of a wide variety of microorganisms, including fastidious bacterial species. Eugon media was developed to obtain eugonic (luxuriant) growth of fastidious microorganisms. The unenriched media supports rapid growth of lactobacilli associated with cured meat products, dairy products and other food. The high concentration of Dextrose is the energy source for rapid growth of bacteria. L-cystine and sodium sulfite are added to stimulate growth. Sodium chloride maintains the osmotic balance of the media. The high carbohydrate content along with high sulfur (cystine) content improves growth with chromogenicity. Colony morphology and color are species specific hours at 35 37º C Candida albicans ATCC 10231, 48 hours at 25 30º C Not undertaken. Eugon Broth: A pure culture of Escherichia coli ATCC appears on Eugon Broth with typical white-creamy to opaque colonies. to ph 7.0 ± 0.2 Pancreatic Digest of Casein Papaic Digest of Soybean Meal Sodium Chloride l-cystine Sodium Sulfite Dextrose g 0.3 g 0.2 g 5.5 g Organism Coloring E. coli ATCC White, cream to opaque colonies Order information Eugon Media Ampouled Media 2 ml Bottled broth 50 ml * 43

9 Media HPC Broth with TTC Products HPC Broth with TTC Used for heterotrophic plate counts in the examination, for example, of potable water, dairy products and swimming pools. HPC is used to determine total count at incubation temperatures of 35º C. All bacteria develop on HPC with indicator media and produce a red color as a result of the precipitation of formazan following the reduction of 2,3,5- triphenyltetrazolium chloride (TTC) by bacteria. incubate for hours at 35º C. Not undertaken. 14 days plated sterility test. HPC with Indicator: Escherichia coli ATCC shows a typical red to pink coloring. to ph 7.1 ± 0.2 Colonies that develop on HPC are counted as total heterotrophic counts. Specific colony identification should be done using standard microbiological identification techniques. Peptone Gelatin Glycerol TTC, 1% solution 30.0 g ml 10.0 ml Characteristics Coloring E. coli ATCC Growth Pink to red E. faecalis ATCC Growth Pink to red S. aureus ATCC Growth Pink to red Order information HPC with TTC Ampouled Media 2 ml

10 Products Media KF-Streptococcus Broth KF-Streptococcus Broth Selected media for the isolation and enumeration of fecal streptococci KF-Streptococcus Broth is selective for the determination of fecal streptococci in polluted surface waters. Maltose and lactose are fermentable carbohydrates, sodium azide is the selective agent and brom cresol purple is the indicator dye. Identification of fecal streptococci has to be undertaken using conventional microbiology techniques. Characteristics Coloring E. faecalis ATCC Growth Red E. faecalis ATCC Growth Red E. aerogenes ATCC inhibited E. coli ATCC inhibited Enterococcus faecalis ATCC 19433, incubated hours at 35 C. incubated hours at 35 C. 14 days plated sterility test. KF-Streptococcus Broth: Pure culture of Enterococcus faecalis ATCC develops on this media a typical red colony coloring. to ph 7.2 ± 0.2 Peptone Yeast extract Sodium chloride Sodium glycerophosphate Maltose Lactose Sodium azide Brom cresol purple TTC 1% solution 20.0 g 1.0 g 0.4 g 15 mg 10.0 ml Order information KF-Streptococcus Media Ampouled Media 2 ml Bottled broth 50 ml * 100 ml * 500 ml * 45

11 Media Lauryl Sulfate or Lauryl Tryptose Broth Products Lauryl Sulfate or Lauryl Tryptose Broth Used for the detection of coliform organisms in water, wastewater and foods. This media was developed for the detection of coliform organisms by the American Public Health Association (APHA). It is now the standard medium of choice in the presumptive phase of the standard coliform MPN test for the microbiological examination of water. Lactose acts as a source of fermentable carbohydrates for coliforms. The fermentation of lactose with gas formation is a presumptive test for coliforms. Sodium lauryl sulfate inhibits the growth of organisms other than coliforms. Escherichia coli ATCC incubation for hours at 35º C. Enterococcus faecalis ATCC incubated at 35º C for hours = no growth. Sterility test: Lauryl Sulfate Broth: Pure culture of Escherichia coli ATCC cultivated on this media shows normal growth with gas formation. to ph 6.8 ± 0.2 Sodium lauryl sulfate Pancreatic Digest of Casein Lactose Dipotassium phosphate Monopotassium phosphate Sodium chloride 0.1 g 20.0 g 2.75 g 2.75 g Characteristics E. coli ATCC Growth/gas E. aerogenes ATCC Growth/gas S. faecalis ATCC inhibited Order information Lauryl Sulfate or Lauryl Tryptose Media Lauryl Sulfate or Lauryl Tryptose Broth 9 ml vials

12 Products Media MacConkey with MUG MacConkey with MUG Used in the presumptive phase for the presence of coliform bacteria in water and foods. Selective to the detection of gram-negative lactose fermenting bacilli. This medium largely complies with the European Pharmacopeia. MacConkey Broth was originally developed by MacCONKEY and HILL (1901) and the formulation for the agar was modified by MacConkey in MacConkey developed this medium for the cultivation of enteric pathogens and coliforms. It contains bile salts, which inhibit certain gram-negative bacteria, and crystal violet, which inhibits grampositive organisms such as enterococci and staphylococci. Combination of the lactose and neutral red indicator to produce pink to red colored colonies indicates an isolate with the ability to ferment lactose. Non-lactose fermenting organisms are colorless. The addition of MUG (4-Methylumbellifryl-β-Dglucuronide), which is a fluorogenic enzyme, allows the media to selectively identify Escherichia coli. MUG is hydrolyzed by the Escherichia coli specific enzyme β-glucuronidase to release 4-Methylumbellifone which fluoresces under ultraviolet light. Fluorescence under UV light is specific for the presence of Escherichia coli. Lactose fermenting organisms grow as pink to red colonies. Characteristics Coloring E. faecalis ATCC inhibited S. typhimurium ATCC Growth Clear E. coli ATCC Growth/fluorescence incubated at 35º C for 24 hours. Checked for florescence at 366 nm. Not undertaken. Sterility. MacConkey with MUG: Escherichia coli ATCC forms as lactose-fermenting organisms pink to red colonies. Make to 1 liter and adjust ph to 7.1 Casein Peptone 17.0 g Proteose Peptone 3.0 g Lactose 10.0g Bile Salts #3 1.5 g Sodium Chloride Neutral Red 30 mg Crystal Violet 0.1 mg MUG (4-Methylumbelliferylβ-D-glucuronide) 0.1 g Additional information: MacConkey Broth is a modification of the original bile salt broth where bromo cresol purple is used in place of neutral red or litmus as the indicator. Bile salts replace 0.5 % sodium taurocholate in the origin formulation. The addition of MUG (4-Methylumbellifryl-(-D-glucuronide), which is a fluorogenic enzyme, allows the media to selectively identify Escherichia coli. MUG is hydrolyzed by the Escherichia coli specific enzyme (β-glucuronidase to release 4-Methylumbellifone which fluoresces under ultraviolet light). Historical background: MacConkey Broth is a modification of the formula given by MacConkey which corresponds to the alternative formulation recommended by the World Health Organization. The medium is used for the presumptive determination of the presence of coliform organisms (gram-negative, lactose fermenting bacilli) in water and milk as well as other materials. Presence of lactose fermenting organisms is detected by the change of color of the medium (from purple to yellow) after inoculation and incubation.the original formula of MacConkey called for use of litmus as an indicator of acid reaction. In later investigations neutral red was found to be a more suitable indicator. More recently, Childe and Allen demonstrated an inhibitory effect on the growth of Escherichia coli by neutral red in this medium. Bromcresol purple is not only less inhibitory, but also gives a more clear cut indication of acid reaction. Order information MacConkey with MUG Ampouled Media 2 ml

13 Media Mannitol Salt Broth Products Mannitol Salt Broth Used for the selective isolation and enumeration of staphylococci. In addition the medium complies with the recommendations in the United States Pharmacopeia. Because of the amount of peptones and beef extract, Mannitol Salt is a nutrient rich medium. Most bacteria (other than staphylococci) are inhibited by the high concentration of sodium chloride. capable of fermenting mannitol e.g. Staphylococcus aureus, cause a ph change in the media. With phenol red as the ph indicator the colonies appear with a yellow coloration. Typical pathogenic staphylococci ferment mannitol and form yellow colonies with yellow zones, while typical non-pathogenic staphylococci do not ferment mannitol and form red colonies. Staphylococcus aureus ATCC 25923, hours at 35º C hours at 35º C. Appearance of Colonies Surrounded by bright yellow zones, abundant growth pink to red colonies growth is usually poorer Microorganisms Mannitol-positive: S. aureus Mannitol-negative: S. epidermis and others Mannitol Salt Broth: Staphylococcus aureus ATCC forms typical yellow colonies with zones on this media. Indication for Mannitol positive organisms. to ph 7.4 ± 0.2 Beef Extract Pancreatic Digest of Casein Peptic Digest of Animal Tissue Sodium Chloride d-mannitol Phenol Red 1.0 g 7 25 mg Characteristics Coloring S. aureus ATCC Growth Yellow with yellow zones S. epidermidis ATCC Growth Red with no zone E. coli ATCC inhibited E. aerogenes ATCC inhibited Historical background: The tolerance of Staphylococcus aureus to high concentrations of sodium chloride was reported by Koch in In 1945, Chapman described a formulation which incorporated 7.5 % sodium chloride in phenol red mannitol agar to successfully cultivate pathogenic staphylococci. These coagulase positive organisms grew in large colonies surrounded by yellow zones. Non-pathogenic staphylococci are usually less luxuriant after the 36 hour incubation period recommended by Chapman. Mannitol Salt Broth and Agar is recommended for the enumeration of staphylococci in food and dairy products by the American Public Health Association. Order information Mannitol Salt Media Ampouled Media 2 ml

14 Products Media Membrane Lauryl Sulfate Broth Membrane Lauryl Sulfate Broth For the presumptive identification of coliforms and Escherichia coli in water and drinking water. Prepared to the formula published in Journal of Hygiene (PHLS/SCA). This media was developed for the detection of coliform organisms and is now the media of choice for the enumeration of total coliforms and Escherichia coli in the United Kingdom. This media replaced membrane enriched broth containing 0.4 % Teepol 610. To use membrane lauryl sulphate broth for the identification of thermotolerant coliforms incubate at 30º C for 4 hours then incubate for further 14 hours at 44º C. Yellow colonies are counted as presumptive coliforms which require confirmation. Characteristics Coloring 37 C E. coli ATCC Growth Yellow E. aerogenes ATCC Growth Yellow E. faecalis ATCC Growth Pink or colorless This picture shows a mix culture on Membrane Lauryl Sulphate Broth incubated at 37 C. like Escherichia coli ATCC and Enterobacter aerogenes ATCC form yellow colonies whereas Enterococcus faecalis ATCC appears as pink colonies. Lactose acts as a source of fermentable carbohydrates for coliforms. Phenol red acts as an indicator of acidity as a result of coliform metabolism. Incubate for 4 hours at 30º C then increase the temperature to 37º C and incubate for further 14 hours. Yellow colonies are counted as presumptive coliforms which require confirmation. Pink or colorless colonies are not counted as coliforms. Characteristics Coloring 44 C E. coli ATCC Growth Yellow E. aerogenes ATCC Inhibited Inhibited E. faecalis ATCC Inhibited Inhibited Escherichia coli ATCC incubation for 4 hours at 30º C then for 14 hours at 37º C. to ph 7.4 ± 0.2 Peptone Yeast extract Lactose Phenol red (0.4% solution) Sodium lauryl sulfate 40.0 g 6.0 g 30.0 g 50.0 ml 1.0 g Not undertaken. Order information Membrane Lauryl Sulfate Media Ampouled Media 2 ml

15 Media M-Endo Coliform Broth Products M-Endo Coliform Broth M-Endo Broth is used for the enumeration of coliforms by membrane filtration. It is used to differentiate lactose from non-lactose fermenting intestinal organisms and as a presumptive test for coliforms. M-Endo is a red colored media, which needs to be stored in the dark to prevent discoloration of the media. Gram-positive bacteria are inhibited on this media by the desoxycholate and lauryl sulfate. The addition of ethanol increases the antibacterial nature of the formulation. Lactose fermenting organisms form aldehydes, which react with Schiffs reagent (basic fuchsin and sodium sulfite) to give red colored zones around the colonies. Coliform colonies are therefore red with a characteristic metallic sheen. Production of both acid and aldehyde by lactose fermenters, such as Escherichia coli, produce deep red colonies that color the surrounding medium and have a green metallic sheen. Non-lactose fermenters form colorless, translucent colonies. Characteristics Coloring E. coli ATCC Growth Red with green metallic sheen E. aerogenes ATCC Growth Red colonies with or without green metallic sheen P. aeruginosa ATCC Growth Pink to colorless S. aureus ATCC Marked to complete inhibition 24 hours at 35º C. Pseudomonas aeruginosa ATCC 10145, 24 hours at 35º C. M-Endo Broth: Mixed culture of Escherichia coli ATCC 25922: (red colonies with green metallic sheen) and Pseudomonas aeruginosa ATCC (pink to colorless). to ph 7.2 ± 0.2 Peptones (equal parts digests of animal tissue and casein) Peptic Digest of Animal Tissue Pancreatic Digest of Casein Yeast extract 1.5 g Lactose 12.5 g Sodium chloride Dipotassium phosphate g Monopotassium phosphate g Sodium lauryl sulfate 50 mg Sodium desoxycholate 0.1 g Sodium sulfite 2.1 g Basic fuchsin 1.05 g 95 % Ethanol 30.0 ml Order information M-Endo Ampouled Media 2 ml Bottled broth 50 ml, Screw cap ml, Septa cap

16 Products Media M-FC Broth M-FC Broth M-FC media (membrane Fecal Coliform media) is used for the detection of fecal coliforms as an index of water pollution. Allows the development of fecal coliforms at elevated temperatures (44.5 C). Bile salts included in the medium inhibit the growth of gram-positive bacteria. Fecal coliforms ferment lactose at elevated temperatures and produce blue colonies. Other organisms form grey to cream colonies. Characteristics Coloring E. coli ATCC Growth Blue E. aerogenes ATCC Growth Gray to cream E. faecalis ATCC inhibited incubated 24 hours at 44.5º C. Enterobacter aerogenes ATCC 13048, incubated 24 hours at 44.5º C. M-FC-Media: A pure culture of Escherichia coli ATCC shows a typical blue coloring on m-fc Broth. to ph 7.4 ± 0.2 Tryptose Peptone No. 3 Yeast extract Sodium chloride Lactose Bile salts Aniline blue g 12.5 g 1.5 g 0.1 g Order information M-FC-Media M-FC-Media: A cultivated mix culture indicates lactose fermenters as blue colonies whereas nonlactose fermenters form grey to creamy colonies e.g. Enterobacter aerogenes ATCC Ampouled Media 2 ml Bottled broth 50 ml * 100 ml * 500 ml * 51

17 Media M-FC with Rosolic Acid Products M-FC with Rosolic Acid M-FC Broth with Rosolic Acid is used for the detection of fecal coliforms by the membrane filtration technique. M-FC with Rosolic Acid acts and functions in the same way as m-fc Broth. Rosolic acid inhibits bacterial growth in general, except for fecal coliforms. Bile salts inhibit non-enteric bacteria. Aniline blue indicates the ability of fecal coliforms to ferment lactose to acid that causes a ph change in the medium. Other organisms form grey to cream colonies. Characteristics Coloring E. coli ATCC Growth Blue E. aerogenes ATCC Growth Gray to cream E. faecalis ATCC inhibited 24 hours at 44,5º C. Enterobacter aerogenes ATCC 13048, 24 hours at 44.5º C. M-FC with Rosolic Acid: Escherichia coli ATCC forms blue colonies whereas non-lactose fermenters appears as grey colonies. to ph 7.4 Tryptose Peptone No. 3 Yeast extract Sodium chloride Lactose Bile salts Aniline blue Rosolic Acid 1% 3.0 g 12.5 g 1.5 g 0.1 g 10.0 ml Additional information: Fecal coliform (FC) Medium for the membrane filtration technique was described by Geldereich et al. in It was the first membrane filtration technique to be incubated at 44.5 ± 0.2. Order information m-fc with Rosolic Acid Ampouled Media 2 ml Bottled broth 50 ml * 52

18 Products Media M-Green Select Broth M-Green Select Broth Used for the enumeration of yeasts and mold in soft drinks and fruit juices. M-Green Select Broth is an improved modification of the liquid medium, m- Green Yeast and Mold Broth and was developed to improve efficiency of detection and enumeration of fungi in sugar based drinks using the membrane filtration method. This medium has a low ph, which inhibits bacterial growth. The addition of Chloramphenicol further inhibits the growth of bacteria to allow for the development and enumeration of yeast and mold. The addition of bromocresol green, which diffuses into fungal colonies as an alkaline reaction, allows them to be easily identified. Metabolic by-products from the developing colonies diffuse into the surrounding medium, further reducing the ph which aids in the inhibition of bacterial growth, but also produces an acid reaction which causes residual bromocresol green to change to yellow. Green opaque colonies against a yellow background indicative of the growth of yeast. Mold colonies are green and filamentous. Candida albicans ATCC 10231, incubated 48 hours at 25 30º C. incubated 48 hours at 35º C. M-Green Select Media: Ideal for the enumeration of yeasts & mold. to ph 4.6 ± 0.2 Dipeptone Yeast extract Dextrose Magnesium sulfate Potassium phosphate Diastase Thiamine Bromocresol green Chloramphenicol, 1% Solution 9.0 g 50.0 g 2.1 g 2.0 g 50 mg 50 mg 26 mg 8.5 ml Characteristics E. coli ATCC Partial to marked inhibition S. cerevisiae ATCC 4098 Growth C. albicans ATCC Growth Order information M-Green Select Media Ampouled Media 2 ml Bottled broth 50 ml * 53

19 Media M-Green Yeast and Mold Products M-Green Yeast and Mold Used for the enumeration of yeast and mold in soft drinks and fruit juices. M-Green is an improved modification of the liquid medium, m-yeast and Mold Broth and was developed to improve efficiency of detection and enumeration of fungi in sugar based drinks using the membrane filtration method. This medium has a low ph, which inhibits bacterial growth. The addition of bromocresol green, which diffuses into fungal colonies as an alkaline reaction, allows them to be easily identified. Metabolic by-products from the developing colonies diffuse into the surrounding medium, further reducing the ph which aids in the inhibition of bacterial growth, but also produces an acid reaction which causes residual bromocresol green to change to yellow. Green opaque colonies against a yellow background are indicative of the growth of yeast. Mold colonies are green and filamentous. Examples of detected organisms: Candida albicans ATCC 10231, 48 hours at 25 30º C. Not undertaken. M-Green Yeast & Mold Broth: Typical growth of Candida albicans ATCC on a black membrane. to ph 4.6 ± 0.2 Dipeptone Yeast extract Dextrose Magnesium sulfate Potassium phosphate Diastase Thiamine Bromocresol green 9.0 g 50.0 g 2.1 g 2.0 g 50 mg 50 mg 26 mg Order information M-Green Yeast & Mold Ampouled Media 2 ml Bottled broth 50 ml * 100 ml * 500 ml * Bottled agar 100 ml

20 Products Media MI Broth and MI Agar MI Broth and MI Agar Used for the simultaneous detection of total coliforms and Escherichia coli in water according to the Surface Water Treatment Rule (USEPA) and the Total Coliform Rule (USEPA) MI Broth detects the presence of coliform bacteria by the production of β-galactosidase, which cleaves the substrate MUGal to produce 4-Methylumbelliferone, which fluoresces on exposure to UV light. Non-coliforms do not produce this enzyme and therefore do not fluoresce on the medium. Escherichia coli is detected by the compound IBDG. The β-glucuronidase produced by Escherichia coli cleaves the substrate to produce a blue indigo color in the colonies. As Escherichia coli is also a total coliform, and also produces β-galactosidase it will also fluoresce. The antibiotic cefsulodin is added to inhibit the growth of gram-positive bacteria and some non-coliform gramnegative bacteria that can cause false positive reactions. MIBlue is specially developed for the food industry. Fluorescent blue colonies are Escherichia coli. Colonies that demonstrate blue/white fluorescence where the colonies are clear, cream or pale yellow in color are other coliform organisms. Clear colonies that do not fluoresce are noncoliforms. Characteristics Coloring E. coli ATCC Growth Blue with fluorescence E. aerogenes ATCC Growth Yellow with fluorescence P. aeruginosa ATCC Complete inhibition hours at 35º C. Enterobacter aerogenes ATCC 13048, hours at 35º C. Pseudomonas aeruginosa ATCC 10145, 24 hours at 35º C. MI-Media: Pure Culture of Escherichia coli ATCC with UV light. Per liter of Water adjusted to ph 6.95 ± 0.2 Protease peptone Yeast extract β-d-lactose MUGal NaCl K2HPO4 KH2PO4 Sodium lauryl sulfate Sodium desoxycholate IBDG Cefsulodin (Agar) 3.0 g 1.0 g 0.1 g 7.5 g 3.3 g 1.0 g 0.2 g 0.1 g 0.32 g 5 mg 1 Order information MI Media MI-Broth Media: Mixed Culture with Escherichia coli ATCC and Enterobacter aerogenes ATCC without UV. Ampouled Media 2 ml Bottled broth 50 ml Bottled agar 50 ml MIBlue * 55

21 Media MRS Broth Products MRS Broth Used for the isolation and cultivation of lactobacilli. MRS Broth complies with the German DIN-Norm and the International Standard ISO for the detection of lactosein meat and to the regulations acc. to 35 LMBG (06.00/35) for the detection in meat. MRS medium supports luxuriant growth of all lactobacilli, even the slow growing species. Lactobaccili appear as white colonies. Pediococcus and Leuconostoc may also grow on MRS. Lactobacillus plantarum ATCC 8014 Lactobacillus fermentum ATCC 9338 incubated at 35º C for hours. Not undertaken. MRS Media; Pure Culture of Lactobacillus plantarum ATCC to ph 6.2 ± 0.2 Pancreatic Digest of Casein Beef extract Yeast extract Dextrose Dipotassium Phosphate Polysorbate 80 Ammonium citrate Sodium acetate Magnesium sulfate Manganese sulfate 8.0 g 4.0 g 20.0 g 2.0 g 1.0 g 2.0 g 0.2 g 50 mg Additional information: The media contains special growth factors for lactobacilli like polysorbate, acetate, magnesium and manganese. These compounds are known as rich nutrient base. On a very low degree of selectivity, Pediococcus and Leuconostoc species and other secondary bacteria can be cultivated. Historical background: MRS Agar was developed by de Man, Rogosa & Sharper for the cultivation of Lactobacillus species. The medium will effectively support the growth of many strains of lactobacilli that do not generally grow as well as other media designed for this purpose. Due to the nutritional factors and the neutral ph other organisms which are not fastidious will grow on the medium. Lactobacillus will grow well on the surface of the medium as well as in deep culture preparations. Although enrichment with carbon dioxide is unnecessary with this medium, the atmosphere must be kept fairly moist Order information MRS Media Ampouled Media 2 ml Bottled broth 9 ml * 56

22 Products Media M TGE Total Count Media M TGE Total Count Media Used for the non-selective development and enumeration of all aerobic bacteria. All bacteria develop on TGE media and produce a range of different colored and sized colonies. Identification of bacteria should be undertaken by using traditional microbiology techniques following initial colony development. For quality control two typical organisms are detected and enumerated with m TGE Total Count Media: Characteristics Coloring E. coli ATCC Growth Yellow to cream S. aureus ATCC Growth Yellow to cream 48 hours at 35º C. Not undertaken. Pure culture of Escherichia coli ATCC on m-tge Total Count Media. Per liter of water and adjusted to ph 7.0 ± 0.2 Pancreatic Digest of Casein Yeast extract Dextrose 2.0 g Order information M-TGE Total Count M-TGE Total Count Media with a mixed culture of Escherichia coli ATCC and Staphylococcus aureus ATCC Ampouled Media 2 ml Bottled broth 50 ml * 100 ml * 300 ml * 500 ml * 57

23 Media Orange Serum Media Products Orange Serum Media Used for the isolation and enumeration of organisms associated with the spoilage of citrus juices. known to grow in single strength and concentrated juices are lactic acid and acetic acid bacteria and yeast. Lactobacilli, Leuconostoc and yeast have all been identified as spoilage organisms by numerous authors. Orange serum at ph 5.4 to 5.6 has been reported to yield maximum counts of all types of spoilage organisms in mixed cultures, and in single culture comparison tests. Lactobacillus fermentum ATCC 9338, 48 hours at 35º C. Candida albicans ATCC 10231, 48 hours at 25 30º C. Not undertaken. Orange Serum Media agar is especially selective for microorganisms which prefer low ph conditions e.g. Lactobacillaceae and some yeast e.g. Candida. Per liter of water, adjusted to ph 5.6 ± 0.2 The low ph of the test solution usually prohibits the development of other microorganisms which cannot survive low ph conditions. Therefore developing colonies are presumed to be problematic organisms. Orange serum Yeast extract Tryptone Dextrose Dipotassium phosphate 3.0 g 4.0 g 2.5 g Order information Orange Serum Ampouled Media 2 ml Bottled Agar 100 ml

24 Products Media Potato Dextrose Broth and Agar Media Potato Dextrose Broth and Agar Media Recommended for culturing and enumerating yeast and mold. Potato Dextrose Broth complies with the recommendations of the American Public Health Association for food and the USP. Potato Dextrose Broth is recommended in Standard Methods as the media that gives the most consistent and highest counts for the recoveries of yeast and mold in dairy products. The inclusion of potato extract encourages the growth and development of fungi. Sterile Tartaric Acid may be added to low the ph to 3.5 ± 0.2 to further inhibit the growth of conflicting bacteria. Yeast, mold and acid tolerant bacteria grow well on Potato Dextrose Broth. Characteristics Coloring C. albicans ATCC Growth White, creamy S. cerevisiae ATCC 4098 Growth White, creamy Candida albicans ATCC 10231, incubated at 25 30º C for 48 hours. Not undertaken. Potato Dextrose Media: Pure culture of Candida albicans ATCC to ph 5.1 ± 0.2 Potato infusion Dextrose For agar add: Agar 4.0 g 20.0 g 1 Additional information: In general carbohydrate and potato infusion promote the growth of yeast and mold. Additionally the low ph value has a positive effect by partially inhibiting the growth of accompanying bacterial flora. If the medium is used for fungal counts, the ph should be adjusted to approximately 3.5. Fungi grow on this medium to develop typical morphology. Historical background: Potato Dextrose Broth is commonly used in slide culture preparations of fungi to stimulate sporulation and to enhance growth of poorly sporulating mycelia. It has been used for cultivation and isolation of mold and yeast in dairy and food products as recommended by the American Public Health Association and for maintenance of stock cultures of geophilic dermatophytes. Order information Potato Dextrose Media Ampouled Media 2 ml Bottled broth 50 ml * 100 ml * 500 ml * Bottled agar 23 ml tube * 100 ml * 500 ml * 1000 ml * 59

25 Media Pseudomonas Broth Products Pseudomonas Broth Used for the isolation of Pseudomonas and differentiating Pseudomonas aeruginosa from other pseudomonads based on pigment formation. Pseudomonas Broth complies with the formulation recommendations of the United States Pharmacopeia and to the specifications in the DIN Norm (examination of water). Pseudomonas aeruginosa is characterised by the production of pyocyanin (a blue green, water soluble, nonflourescent, phenazine pigment), which is stimulated by the inclusion of magnesium chloride and potassium sulfate in the broth. Irgasan, an antimicrobial agent, selectively inhibits gram positive and gram negative bacteria other than pseudomonads. Glycerol serves as both an energy source and helps in the promotion of pyocyanin. Development of a green to blue green pigmentation surrounding colonies is positive for Pseudomonas aeruginosa. Other pseudomonads develop as clear to amber yellow colonies. Non-pseudomonads are suppressed. Characteristics Coloring P. aeroginosa ATCC Growth Blue to blue-green P. aeroginosa ATCC Growth Blue to blue-green E. coli ATCC inhibited Pseudomonas aeruginosa ATCC 10145, hours at 35º C. 24 hours at 35º C. Pseudomonas Media: Typical growth of Pseudomonas aeroginosa ATCC Per liter of Water adjusted to ph 7.0 ± 0.2 Pancreatic Digest of Casein Magnesium chloride Potassium sulfate Irgasan Glycerol 20.0 g 1.4 g 0.25 g 20.0 ml Additional information: Identification of most Pseudomonas strains can be obtained by their different pigmentation according to the compounds used in the media. Some of the strains can only synthesize pyocyanin, some form only fluorescein and others form both pigments. Pseudomonas aeruginosa ATCC is characterised by the production of pyocyanin (a blue green, water soluble, nonflourescent, phenazine pigment), which is stimulated by the inclusion of magnesium chloride and potassium sulfate in the broth. The same Pseudomonas aeruginosa strain appears on Pseudomonas Broth as colonies surrounded by a yellow to greenish-yellow zone when different types of peptone are added and magnesium chloride and potassium sulphate are omitted. BLAZEVIC et al. (1973), noted that some Pseudomonas aeruginosa strains atypically appear as pyocyanin-negative, fluorescein-positive and for that reason it is possible to differentiate them from Pseudomonas fluorescens and Pseudomonas putida. Furthermore BRODSKY and NIXON (1973) showed that cultivation on MacCONKEY agar can be used for a simple and rapid differentiation of these strains. Order information Pseudomonas Media Ampouled Media 2 ml Bottled broth 50 ml * 100 ml * 500 ml * Bottled agar 50 ml * 100 m * 500 ml * 60

26 Products Media R2 Broth and R2 Agar R2 Broth and R2 Agar Used for Heterotrophic plate counts in the examination of potable water. R2 broth can be used to determine heterotrophic plate count at 35ºC. When incubated at lower temperatures (25 30º C) for longer periods of hours it can also be used to recover environmentally stressed organisms, or those that are chlorine tolerant. Heterotrophic plate counts are not the same as standard plate counts and a media such as TGE should be run in combination with R2. Characteristics E. coli ATCC Growth E. faecalis ATCC Growth S. aureus ATCC Growth incubation for not less than 72 hours at 35 C. Not applicable. Sample of tap water on R2 Media. to ph 7.2 ± 0.2 Yeast extract Peptones Acid hydrolysate of casein Dextrose Soluble starch Dipotassium phosphate Magnesium sulfate (anhydrous) Sodium pyruvate 0.5 g 0.5 g 0.5 g 0.5 g 0.5 g 0.3 g 24 mg 0.3 g For agar add: Agar 1 Order information R2 Media Ampouled Media 2 ml Bottled agar 15 ml tube ml ml * 61

27 Media Sabouraud Dextrose Broth Products Sabouraud Dextrose Broth The medium is used for the quantitative identification of yeast and mold. Description Peptone in the media is used as a nitrogen source for the development of fungi. Dextrose acts as an energy source for the growth of microorganisms. The low ph is favorable for the development of fungi, especially dermatophytes, but at the same time inhibits the development of contaminating bacteria from clinical specimens. Growth is interpreted as yeast/mold. Subculturing and species/group specific identification is required. Candida albicans ATCC incubated for 72 hours at 25 30º C. Not undertaken. Sabouraud Dextrose Broth is especially selective for yeast and mold due to the low ph conditions. to ph 5.6 ± 0.2 Peptone Dextrose For agar add: Agar 20.0 g 1 Characteristics Coloring C. albicans ATCC Growth Off-white, creamy E. coli ATCC Growth Off-white, creamy S. cerevisiae ATCC 9763 Growth Off-white Historical background: Sabouraud Dextrose Broth is a modification of dextrose agar described by Sabouraud in 1892 for identification of fungi based on their cultural characteristics. The medium depended solely upon its acid ph for suppression of bacteria. Sabouraud Dextrose Agar, Emmons is a modification with a neutral ph and reduced dextrose concentration. Historically, the inhibitory action of Sabouraud Dextrose Agar on microorganisms other than fungi was enhanced by the addition of agents such as tellurite, copper sulfate, bile salts, dyes and antibiotics. Sabouraud Dextrose Broth (Fluid Sabouraud Medium) is the liquid counterpart of the agar prepared according to the formulation specified in the U.S. Pharmacopoeia and National Formulary for sterility testing of pharmaceutical products. Used in the quantitative identification of yeast and mold. Order information Sabouraud Dextrose Media Ampouled Media 2 ml Bottled broth 50 ml * 100 ml * 500 ml * Bottled agar 50 ml * 100 ml * 500 ml * 62

28 Products Media Sabouraud 4 % Dextrose Broth Sabouraud 4 % Dextrose Broth Used in the quantitative identification of yeast, mold and acidophilic microorganisms.this culture medium complies with the recommendations of the United States Pharmacopeia and the European Pharmacopeia. Peptone in the media is used as a nitrogen source for the development of fungi. Dextrose acts as an energy source for the growth of microorganisms. The low ph is favorable for the development of fungi, especially dermatophytes, but at the same time inhibits the development of contaminating bacteria from clinical specimens. The use of Sabouraud media has been documented for establishing the microbial concentration in cosmetics and for the mycological evaluation of food. Growth is interpreted as yeast/mold. Sub-culturing and species/group specific identification is required. Candida albicans ATCC 10231, incubated 72 hours at 25 30º C. Not undertaken. Sabouraud 4 % Dextrose Media: Pure culture of Candida albicans ATCC to ph 5.6 ± 0.2 Peptone Dextrose 40.0 g Characteristics C. albicans ATCC Growth S. cerevisiae ATCC 9763 Growth Order information Sabouraud 4% Dextrose Media Ampouled Media 2 ml

29 Media Standard Methods Agar Products Standard Methods Agar Used for obtaining microbiological plate counts from milk and dairy products, foods, water and other materials of sanitary importance. The formulation of this medium follows the demands of the Standard Methods for the Examination of Water and Wastewater, the American Public Health Association, the American Water Works Association and the Water Pollution Control Federation in 1992 and the Standard Methods for the Examination of Dairy Products of the American Public Health Association (1985). All bacteria develop on Standard Methods and produce a range of different colored and sized colonies. It is not possible using Standard Methods Agar to presumptively identify any bacteria. Identification can only be undertaken using traditional microbiology techniques following initial colony development. Characteristics Coloring E. coli ATCC Growth Yellow to cream S. aureus ATCC Growth Yellow to cream incubated 48 hours at 35º C. Not undertaken. Historical background: Standard Methods Agar is a modification of Bowers and Hucker Tryptone Glucose Skim Milk Agar. It was shown by Yale to be more effective in plate count procedures on milk and dairy products. The American Public Health Association in their Standard Methods for the Examination of Dairy Products recommends the use of Standard Methods Agar for performance of the Standard Plate Count on dairy products. Standard Methods Agar is the same formulation as Plate Count Agar recommended by the APHA for use in the Standard Plate Count on water. Standard Methods Agar with membrane filter and incubated with a pure culture of Escherichia coli ATCC to ph 7.0 ± 0.2 Pancreatic Digest of Casein Yeast extract Dextrose Agar 2.5 g 1.0 g 1 Same agar as above but incubated with a mix culture of Escherichia coli ATCC and Staphylococcus aureus ATCC Both form yellow to creamy colonies. Order information Standard Methods Medium Bottled agar 100 ml

30 Products Media Total Count Media with TTC Total Count Media with TTC Total Count Media with Indicator is used for the non-selective development and enumeration of all aerobic bacteria by the membrane filtration method. This media complies with the specifications given by the APHA for the examination of water (1992) and for food (1992). All bacteria develop on Total Count Media with indicator and produce a red color as a result of the precipitation of formazan following the reduction of 2,3,5-triphenyltetrazolium chloride (TTC) by bacteria. The table below shows typical organisms which can be enumerated with this media: It is not possible using Total Count Media with Indicator to presumptively identify any bacteria. Identification can only be undertaken using traditional microbiology techniques following initial colony development. Escherichia coli ATCC incubated hours at 35 C. Not undertaken. Sterility Total Count Media with Indicator: Escherichia coli ATCC and Staphylococcus aureus ATCC can be easily detected according to their red to pink colonies. to ph 7.0 ± 0.2 Pancreatic Digest of Casein Yeast extract Dextrose TTC, 1% solution 2.0 g 10.0 ml Characteristics Coloring E. coli ATCC Growth Pink to red S. aureus ATCC Growth Pink to red P. aeruginosa ATCC Growth Pink to red E. faecalis ATCC Growth Pink to red Order information Total Count Media with Indicator Ampouled Media 2 ml Bottled broth 50 ml * 65

31 Media Trypticase Soy Broth (TSB) Products Trypticase Soy Broth (TSB) Trypticase Soy Broth (TSB) is a general-purpose medium used for the cultivation of most bacteria. TSB broth is a medium that will support the growth of a wide variety of microorganisms including aerobic, facultative and anaerobic bacteria and fungi. When used as a broth adsorbed onto pads TSB will support the development of mixed cultures of bacteria and fungi. When used in sterility test applications turbidity indicates the presence of organisms when compared to an uninoculated control. Positive controls: incubation for hours at 35º C. Not undertaken. Ready to use Tryptic Soy Broth (USP). Not inoculated. to ph 7.3 ± 0.2 Pancreatic digest of casein Papaic digest of soybean meal Sodium chloride Dipotassium phosphate Dextrose 17.0 g 3.0 g 2.5 g 2.5 g Characteristics B. subtilis ATCC 6633 Growth C. albicans ATCC Growth E. coli ATCC Growth S. aureus ATCC Growth Order information Trypticase Soy Media (TSB) Ampoules 2 ml Bottled media 50 ml * 100 ml * 100 ml, Septa Cap * 100 ml * Bottled agar 23 ml * 66

32 Products Media Trypticase Soy Broth Single Strength Trypticase Soy Broth Single Strength General-purpose medium used in qualitative procedures for the cultivation of fastidious and non-fastidious microorganisms. Trypticase Soy Broth Single Strength complies with the demands of the DIN Norm for the detection of Escherichia coli serotype 0157:H7 in foods and FDA-BAM for the isolation of enterohemorrhagic Escherichia coli (EHEC). In addition the media conforms to the formula of the U.S. Pharmacopoeia. When used as a broth adsorbed onto pads TSB will support the development of mixed cultures of bacteria and fungi. When used in sterility test applications turbidity indicates the presence of organisms when compared to an uninoculated control. incubated hours at 35º C. Not undertaken. Trypticase Soy Broth-Single Strength. Not inoculated. to ph 7.3 ± 0.2 Pancreatic digest of casein Papaic digest of soybean meal Sodium chloride Dipotassium phosphate Dextrose 17.0 g 3.0 g 2.5 g 2.5 g Characteristics B. subtilis ATCC 6633 Growth C. albicans ATCC Growth E. coli ATCC Growth S. aureus ATCC Growth Historical background: Trypticase Soy Broth was originally developed for detecting the effectiveness of sulfonamides against microorganisms such as pneumococci without the addition of serum or blood to the medium. Spink and Hamilton demonstrated the suitability of the medium for cultivation of aerobic and facultative microorganisms including Brucella. Garrision and Hedgecock both used the medium to promote the growth of pathogenic fungi. The addition of agar to enhance the growth of anaerobic organisms is described by Mashima and Ellison and Fredette, Auger, and Forget. Sherman and Stark demonstrated that a medium containing 6.5 % NaCl could be used for differentiation of group D enterococci from other streptococci. Order information Trypticase Soy Broth-Single Strength Bottled broth 10 ml, Vials * 100 ml ml, wide mouthed * 67

33 Media Trypticase Soy Broth Double Strength Products Trypticase Soy Broth Double Strength General-purpose medium used in qualitative procedures for the cultivation of fastidious and non-fastidious microorganisms. This media is also recommended for sterility test according to U.S. Pharmacopeia. TSB broth is a medium that will support the growth of a wide variety of microorganisms including aerobic, facultative and anaerobic bacteria and fungi. When used as a broth adsorbed onto pads TSB will support the development of mixed cultures of bacteria and fungi. When used in sterility test applications turbidity indicates the presence of organisms when compared to an uninoculated control. Positive controls: incubated hours at 35º C. Not undertaken. Trypticase Soy Broth (Double Strength). Not inoculated. to ph 7.3 ± 0.2 Pancreatic digest of casein Papaic digest of soybean meal Sodium chloride Dipotassium phosphate Dextrose 34.0 g 6.0 g Characteristics B. subtilis ATCC 6633 Growth C. albicans ATCC Growth E. coli ATCC Growth S. aureus ATCC Growth Trypticase Soy Broth (Double Strength): left vial: control, right vial inoculated with Escherichia coli ATCC Order information Trypticase Soy Broth (Double Strength) Bottled broth 50 ml, W/M bottle ml

34 Products Media Wallerstein Nutrient Broth (WL) and WL Differential Broth (WLD) Wallerstein Nutrient Broth (WL) and WLDifferential Broth (WLD) WL Nutrient broth is used for the determination of microbiological flora in brewing and fermentation processes. WL differential media suppresses the growth of yeast and mold and cultivated bacteria encountered in brewing and industrial fermentation. The medium has the same formulation as WL nutrient media but has 4 mg of Cyclohexamide per liter added. Use of the medium at ph 5.5 and incubation at 25 C will give reliable counts for brewer s yeast. Adjustment of the ph to 6.5 and incubation at 30 C allows for the selective growth of bakers and distillers yeast. Interpretation WL Nutrient Broth: Media will develop all yeast that incubate at the selected temperature and ph range. WL Nutrient Broth Characteristics Escherichia coli ATCC 25922Growth L. fermentum ATCC 9338 Growth S. cerevisiae ATCC 9763 Growth Interpretation WL Differential Broth: When incubated at ph 5.5 estimates of beer cocci and lactic rods can be made under anaerobic conditions. Estimates of acetic acid rods and thermobacteria are made under aerobic conditions. WL Differential Broth Characteristics Escherichia coli ATCC 25922Growth L. fermentum ATCC 9338 Growth S. cerevisiae ATCC 9763 Partial to marked inhibition incubation conditions WL Nutrient: Sacchromyces cerivisiae ATCC 4098, incubated 48 hours at C. incubated hours at 35 C. Not applicable. incubation conditions WL Differential: Lactobacillus plantarum ATCC 8014, incubated 48 hours at 35 C. Sacchromyces cerivisiae ATCC 4098, incubated 48 hours at C. Culture of Sacchromyces cerivisiae ATCC 4098 on Wallerstein Nutrient Broth (WL). to ph 5.5 ± 0.2 Casein Yeast Extract Dextrose Monopotassium phospate Potassium chloride Calcium chloride Magnesium sulfate Ferric chloride Manganese sulfate Bromocresol green Additional Component for WL Differential Broth: Cyclohexamide 4.0 g 50.0 g 550 mg 425 mg 125 mg 125 mg 2.5 mg 2.5 mg 22 mg 4 mg Additional information: WL Nutrient Broth has a ph of 5.5 which is optimal for the enumeration of brewers yeast. In the case of cultivating microorganisms from an alcoholic mash, tomato juice should be added to the medium. WL Differential Media contains additionally 4 mg of Cyclohexamide per liter. This component inhibits the developments of yeast without interfering with the development of bacteria generally encountered in beers. Cyclohexamide in WL Differential Broth is added to suppress yeast and any mold which may be present. 69

35 Media Wallerstein Nutrient Broth (WL) and WL Differential Broth (WLD) Products Historical background: WL Nutrient Agar and WL Differential Agar were developed by GRAY and GREEN (1950), to study the microbial population of worts, beers, liquids, yeast, and other materials used in fermentation control procedures of the brewing industry and other fermentation industries. The WL Nutrient Agar is used for the cultivation and enumeration of yeast. At ph 5.5 brewers yeast will grow unrestricted at 25º C within hours. For growth of bakers and distillers yeast, the ph of the medium should be adjusted to 6.5 with incubation at 30º C for 2 7 days. Where yeast cells are in low number, certain bacteria can be detected. The WL Differential Broth is used to determine bacterial counts. GRAY and GREEN (1950) recommended using the differential medium to culture bacteria normally encountered in brewing; aerobic conditions for acetic acid and thermophilic bacteria, anaerobic conditions for lactic bacteria and beer cocci. Wallerstein Differential Broth (WLD): Cultur of Lactobacillus plantarum ATCC Order information WL Nutrient Media Ampoules 2 ml Order information WL Differential Media Ampoules 2 ml Bottled Media 50 ml * 70