Dehydrated Media and Bases

Specifications

Ingredients : g/L Casein enzymic hydrolysate 10.00 Meat Extract 10.00 Yeast extract 3.00 Dextrose 5.00 Sodium chloride 5.00 Sodium acetate 3.00 Starch, soluble 1.00 L-Cysteine hydrochloride 0.50 Agar 0.50 Final pH (at 25°C) : 6.8 + 0.2 Directions : Suspend 38.00 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 10 lbs pressure (115°C) for 15 minutes. Mix well and dispense as desired. Principle : Reinforced Clostridial Agar contains casein enzymic hydrolysate and Meat Extract B as sources of carbon, nitrogen, vitamins and minerals. Yeast extract supplies B-complex vitamins which stimulate bacterial growth. Dextrose is the carbohydrate source. Sodium chloride maintains the osmotic balance. In low concentrations, soluble starch detoxifies metabolic byproducts. Cysteine hydrochloride is the reducing agent. Sodium acetate acts as a buffer. The small amount of Agar makes the medium semisolid.

Specifications

Ingredients: gms/lit. Peptone (meat or casein) 1.00 Sodium chloride 4.30 Disodium hydrogen phosphate dihydrate 7.23 Potassium dihydrogen phosphate 3.60 Final pH (at 25°C) : 7.0 + 0.2 Directions : Suspend 14.64 grams (the equivalent weight of dehydrated medium per litre) in 1000 ml purified /distilled water. Heat if necessary, to dissolve the medium completely. For preparation of nonfatty products insoluble in water, add 0.1 % w/v Polysorbate 80 to assist the suspension of poorly wettable substances. Dispense in tubes or flasks or as desired and sterilize by autoclaving at 15 lbs pressure 121°C for 15 minutes or as per validated cycle. Principle : Buffered Peptone Water is a pre-enrichment medium designed to help recovery of sub-lethally damaged Salmonellae before transfer to a selective medium. This pre-enrichment medium is free from inhibitors and is well buffered and provides conditions for resuscitation of the cells that have been injured by processes of food preservation. Pre-enrichment in Buffered Peptone Water at 37°C for 18 hours before selection in Tetrathionate Brilliant Green Bile Broth showed superior results compared with direct selection method. The composition of Buffered Peptone Water with NaCl medium is as per IP and EP specifications recommended to dilute the sample for microbial examination. Depending on the amount of fat in the sample to examine the kind and quantity of emulsifying agent to be used. These pre-enrichment media contain peptone as a source of carbon, nitrogen, vitamins and minerals. Sodium chloride maintains the osmotic balance and phosphates buffer the medium. The broth is rich in nutrients and produces high resuscitation rates for sub-lethally injured bacteria and supports intense growth. The phosphate buffer system prevents bacterial damage due to changes in the pH of the medium.

Specifications

Ingredients : gms/lit. Peptone 20.00 Lactose 10.00 Sodium taurocholate 5.00 Sodium Chloride 5.00 Bromo cresol purple 0.01 Final pH (at 25°C) : 7.4+ 0.2 Directions : Suspend 40.01 grams in 1000 ml distilled water. Heat if necessary, to dissolve the medium and distribute into test tubes with inverted Durham tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Principle: Peptone provides nitrogeneous and carbonaceous compounds, long chain amino acids and other essential growth nutrients. Lactose is the fermentable carbohydrate. Sodium taurocholate inhibits gram-positive organisms. Sodium chloride maintains the osmotic balance of the medium. Bromocresol purple is the pH indicator in the medium which turns yellow under acidic condition. Lactose fermentation turn the medium yellow due to the acidity produced on lactose fermentation. The colour change of the dye is observed when the pH of the medium falls below 6.8. Lactose non-fermenting organisms like Salmonella and Shigella do not alter the appearance of the medium.

Specifications

Ingredients : gms/lit. Pancreatic Digest of Casein 17.00 Soya Peptone 3.00 Sodium chloride 5.00 Dipotassium Hydrogen Phosphate 2.50 Glucose 2.50 Bile Salts No.3 1.50 Final pH (at 25°C) : 7.3 + 0.2 Directions : Suspend 31.5 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Distribute 250 ml into flasks and sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50oC and to each 250 ml flask add, under aseptic conditions Novobiocin supplement reconstituted with 2.0 ml of sterile distilled water. Final concentration should be 20.0 mg/liter. Principle : The combination of Pancreatic Digest of Casein and Soya peptone makes these media nutritious by providing amino acids and long chain peptides for the growth of microorganisms. Dextrose and dipotassium phosphate serves as the carbohydrate source and the buffer in the medium. Sodium chloride maintains osmotic balance in both the media.

Specifications

Ingredients : gms/lit. Peptone 20.00 Lactose 10.00 Bile Salts 5.00 Sodium Chloride 5.00 Neutral red 0.075 Final pH (at 25°C) : 7.4+ 0.2 Directions : Suspend 40.07 grams in 1000 ml distilled water. Heat if necessary, to dissolve the medium completely. Distribute into tubes with inverted Durhams tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool the tubes before inoculation. Principle: Peptone provides necessary nitrogen source. Lactose serves as the fermentable carbohydrate source. Sodium chloride maintains the osmotic balance of the cells. The selective action of these media is attributed to the presence of bile salts, which are inhibitory to most species of gram-positive bacteria. Gram-negative bacteria usually grow well on these media and are differentiated by their ability to ferment lactose. The colour change of the medium shown by lactose-fermenters is due to production of acid from lactose and a subsequent colour change of the indicator dye when the pH of the media falls below 6.8. The medium turns pink in case of lactose fermenters and yellow in case of non-lactose-fermenters, due to neutral red. MacConkey Broth , which contains neutral red as an indicator is considered as a standard medium for the primary isolation as well as presumptive identification of coliform-aerogenes group of organisms in food and water.

Specifications

Ingredients : gms/lit. Pancreatic digest of casein 17.00 Papaic digest of soyabean meal 3.00 Sodium chloride 5.00 Dipotassium phosphate 2.50 Dextrose 2.50 Final pH (at 25°C) : 7.3 + 0.2 Directions : Suspend 30 gms. in 1000 ml distilled water. Boil to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Principle : The combination of pancreatic digest of casein and papaic digest of soyabean meal makes the medium nutritious by providing amino acids and long chain peptides for the growth of microorganisms. Dextrose and dibasic potassium phosphate serve as the carbohydrate source and the buffer, respectively in the medium. Sodium chloride maintains the osmotic balance of the medium.

Specifications

Ingredient g/L Pancreatic digest of casein 17.00 Papaic digest of soyabean meal 3.00 Sodium chloride 5.00 Dipotassium phosphate 2.50 Dextrose 2.50 Final pH (at 25°C) : 7.3 + 0.2 Direction Suspend 30 gms. in 1000 ml distilled water. Boil to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Principle The combination of pancreatic digest of casein and papaic digest of soyabean meal makes the medium nutritious by providing amino acids and long chain peptides for the growth of microorganisms. Dextrose and dibasic potassium phosphate serve as the carbohydrate source and the buffer, respectively in the medium. Sodium chloride maintains the osmotic balance of the medium. QC Tests (I) Dehydrated Medium Colour : Cream to light yellow Appearance : Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating) : 7.3 ± 0.2 Colour (post autoclaving/heating) : Light yellow Clarity (post autoclaving/heating) : Clear (III) Q.C. Test Microbiological Cultural characteristics observed after 18 – 48 hours at 35-37°C. MICROORGANISM (ATCC ) GROWTH Escherichia coli (25922) Luxuriant Escherichia coli (8739) Luxuriant Escherichia coli (NTCC9002) Luxuriant Salmonella abony (6017 Luxuriant Salmonella typhimurium (14028) Luxuriant Staphylococcus aureus (6538) Luxuriant Pseudomonas aeruginosa (27853) Luxuriant Pseudomonas aeruginosa (9027) Luxuriant Candida albicans (10231 ) Luxuriant Staphylococcus aureus (25923) Luxuriant Bacillus subtilis (6633) Luxuriant Staphylococcus aureus (6538) Luxuriant Micrococcus luteus (9341) Luxuriant Use A general purpose medium used for cultivation of a wide variety of microorganisms and sterility testing of molds and lower bacteria. Storage Dehydrated medium- below 30°C Prepared medium– Between 2 to 8°C.

Specifications

Ingredient g/L Tryptone 15.00 Soya peptone 5.00 Sodium chloride 5.00 Agar 15.00 Final pH (at 25°C):7.3 + 0.2 Direction : Suspend 40 grams in 1000 ml distilled/purified water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. If desired, aseptically add 5% v/v defibrinated blood in previously cooled medium to 45-50°C for cultivation. Mix well and pour into sterile Petri plates. Principle The medium with addition of blood provides perfectly defined hemolysis zones, while preventing the lysis of erythrocytes due to its sodium chloride content.It's simple and inhibitor-free composition makes it suitable for the detection of antimicrobial agents in the food and other products.The combination of tryptone and soya peptone makes this media nutritious by providing amino acids and long chain peptides for the growth of microorganisms. Sodium chloride maintains the osmotic balance. Type of specimen : Pharmaceutical samples, Clinical samples- urine, faeces, abscess etc. Specimen Collection and Handling: For clinical samples follow appropriate techniques for handling specimens as per established and current guidelines of clinical microbiology. For Pharmaceutical samples, follow appropriate techniques for sample collection,handling and processing as per standard and current guidelines of pharmacopeias. After use, contaminated materials must be sterilized by autoclaving before discarding. QC Tests (I) Dehydrated Medium Colour: Cream to light yellow Appearance: Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating): 7.3 ± 0.2 Colour (post autoclaving/heating): a) Llight yellowb) After addition of5-7%w/v sterile defibrinated blood : Cherry red Clarity (post autoclaving/heating): a) Clear slightly opalescentb) Opaque (III) Q.C. Test Microbiological Cultural characteristics was observed after an incubation for Bacterial at 30-35°C 18-24 hours and for Fungal at 30-35°C <=5days. MICROORGANISM (ATCC ) GROWTH W/ BLOOD HAEMOLYSIS Bacillus subtilis (6633) Luxuriant None Staphylococcus aureus (6538) Luxuriant β Staphylococcus aureus (25923) Luxuriant β Escherichia coli (25922) Luxuriant None Escherichia coli (8739) Luxuriant None Escherichia coli (NCTC9002) Luxuriant None Escherichia coli (11775) Luxuriant None Escherichia coli (NCTC13167) Luxuriant None Pseudomonas aeruginosa (10145) Luxuriant None Pseudomonas aeruginosa (27853) Luxuriant None Pseudomonas aeruginosa (9027) Luxuriant None Salmonella Abony (NCTC6017) Luxuriant None Micrococcus luteus (9341) Luxuriant None Streptococcus pneumoniae (6305) Luxuriant None Salmonella Typhimurium (14028) Luxuriant None Enterococcus faecalis (29212) Luxuriant None Candida albicans (10231) Luxuriant None Candida albicans (2091) Luxuriant None Clostridium perfringenes (13124) Luxuriant None Aspergillus brasiliensis (16404) Good-Luxuriant None Use It is a general-purpose medium used for cultivation of a wide variety of microorganismsfrom clinical and non-clinical samples and for sterility testing inpharmaceutical procedures. Storage Dehydrated medium- below 30°C Prepared medium– Between 2 to 8°C

Specifications

Ingredient g/L Proteose Peptone 5.00 Tryptone 5.00 Yeast extract 5.00 Meat extract B# 5.00 Sodium chloride 20.00 Disodium hydrogen phosphate 9.50 Potassium Dihydrogen phosphate 1.35 Aesculin 1.00 Lithium Chloride 3.00 Nalidixic acid 0.010 Acriflavine hydrochloride (Trypaflavine) 0.0125 # Equivalent to Beef Extract Final pH (at 25°C): 7.2 + 0.2 Direction : Suspend 27.4 gms. in 500ml. distilled water. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to room temperature and add the contents of one vial of Fraser Supplement . Mix well and pour into sterile tubes or plates. Principle This medium contains tryptone, yeast extract and beef extract which provide essential nutrients like carbon and nitrogenous compounds including vitamins, amino acids and trace ingredients. Phosphates provide buffering action to the medium while sodium chloride maintains osmotic equilibrium. Nalidixic acid and acriflavine inhibits the growth of gram – negative and gram - positive organisms respectively except Listeria species. QC Tests (I) Dehydrated Medium Colour: Cream to light yellow Appearance: Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating): 7.2 ± 0.2 Colour (post autoclaving/heating): Yellow coloured solution with bluish tinge Clarity (post autoclaving/heating): Clear to slightly opalescent (III)Q.C. Test Microbiological Cultural characteristics observed after 24-48 hours at 35- 37°C. MICROORGANISM (ATCC) GROWTH ESCULIN HYDROLYSIS* Listeria monocytogenes (19118) Luxuriant + Staphylococcus aureus (25923) Inhibited - Escherichia coli (25922) Inhibited - Enterococcus faecalis (29212) Inhibited - Key: + = blackening of medium * = subcultured on Listeria selective agar Use For selective enrichment of Listeria species as per ISO11290-1:1997 & ISO11133 2:2003 Storage Dehydrated medium - - below 30°C Prepared medium– Between 2 to 8°C.

Specifications

Ingredient (Per vial sufficient for 500 ml medium) Chloramphenicol 50.00 mg Direction Rehydrate the contents of 1 vial aseptically with 2 ml of ethanol. Mix well and aseptically add to 500 ml of sterile, molten SABHI Agar Base / Rose Bengal Chloramphenicol Agar Base / Dichloran Medium Base with Rose Bengal / spergillus Differentiation Medium Base / DG18 Agar Base / DRBC Agar Base. Mix well and pour into sterile petri plates. Use An antibiotic supplement recommended for the selective isolation of yeasts and molds. Storage Between 2 to 8°C.

Specifications

Ingredient g/L Peptone 5.00 Yeast extract 1.00 Ferric citrate 0.10 Sodium chloride 19.45 Magnesium chloride 8.80 Sodium sulphate 3.24 Calcium chloride 1.80 Potassium chloride 0.55 Sodium bicarbonate 0.16 Potassium bromide 0.08 Strontium chloride 0.034 Boric acid 0.022 Sodium silicate 0.004 Sodium fluorate 0.0024 Ammonium nitrate 0.0016 Disodium phosphate 0.008 Agar 15.00 Final pH (at 25°C) : 7.6 + 0.2 Direction Suspend 55.25 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and Pour in sterile Petri plates. Principle This medium consists not only of minerals as sea water but also has peptone and yeast extract as the better sources of nutrients for the marine bacteria. Agar is the solidifying agent. Marine bacteria are of prime importance in food industry and marine life conservation and hence activity of these bacteria can be studied by the use of this medium. QC Tests (I) Dehydrated Medium Colour : Cream to yellow Appearance : Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating) : 7.6 ± 0.2 Colour (post autoclaving/heating) : Yellow Clarity (post autoclaving/heating) : Opalescent (III) Q.C. Test Microbiological Cultural characteristics observed after an incubation at 20-25°C for 40-72 hours . MICROORGANISM (ATCC ) GROWTH Vibrio fischeri (7744) Good –luxuriant Vibrio harveyi (14126) Good –luxuriant Use For cultivation, isolation and enumeration of heterotrophic marine bacteria. Storage Dehydrated medium- below 30°C Prepared medium– Between 2 to 8°C.

Specifications

Ingredient (per vial sufficient for 500 ml medium) Acriflavin hydrochloride 6.00 mg Nalidixic acid 10.00 mg Direction Rehydrate the contents of 1 vial aseptically with 2 ml of 0.2 N Sodium Hydroxide. Mix well and aseptically add to 500 ml sterile Listeria Enrichment Medium Base. Mix well and dispense as desired. Use An antimicrobial supplement recommended for the selective cultivation of Listeria species. Storage Between 2 to 8°C.

Specifications

Ingredient g/L Casein enzymic hydrolysate 5.00 Proteose peptone 5.00 Meat extract B# 5.00 Yeast extract 5.00 Sodium chloride 20.00 Monopotassium dihydrogen phosphate 1.35 Disodium hydrogen phosphate 12.00 Esculin 1.00 Nalidixic acid 0.02 #- Equivalent to Beef extract Final pH (at 25°C) : 7.4 + 0.2 Direction Suspend 54.37 gms.in 1000ml. distilled water.Heat to boiling to dissolve the medium completely.Sterilize by autoclaving at 15 lbspressure (121°C) for 15 minutes. Cool to 50°C and aseptically add filter sterilized solution of acriflavin hydrochloride to a final Concentration of 12 mg per litre of medium or 25 mg per litre as per the requirement. Principle Casein enzymic hydrolysate, proteose peptone, beef extract and yeast extract provide necessary nutrients while esculin offers differential properties to the medium. Nalidixic acid and acriflavin hydrochloride together with higher concentration of phosphate render the medium selective for Listeria, as the gram – negative and gram – positive organisms are inhibited by Nalidixic acid and acriflavin hydrochloride respectively. QC Tests (I) Dehydrated Medium Colour : Cream to yellow Appearance : Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating) : 7.4 ± 0.2 Colour (post autoclaving/heating) : Medium amber Clarity (post autoclaving/heating) : Slightly opalescent (III) Q.C. Test Microbiological Cultural characteristics observed after 24- 48 hours at 35-37°C. MICROORGANISM (ATCC ) GROWTH Listeria monocytogenes (19118 ) Good – luxuriant Escherichia coli (25922) None – poor Staphylococcus aureus (25923) None – poor Use For selective isolation and cultivation of Listeria monocytogenes from clinical specimens. Storage Dehydrated medium andprepared medium– Between 2 to 8°C.

Specifications

Ingredient (100 ml per vial) Egg Yolk 30.00 ml Sterile saline 64.00 ml Sterile 3.5% potassium tellurite solution 6.00 ml Final pH (at 25°C) 7.6±0.2 Direction Warm up the refrigerated Egg Yolk Tellurite Emulsion to 40-45°C. Shake well to attain uniform emulsion (since on refrigeration emulsion has a tendency to form layers or small lumps). Aseptically add 50 ml in 950 ml of sterile, molten Baird Parker Agar Base , Baird Parker Agar w/Sulfate Mix well and pour into sterile petri plates. Use Sterile stabilized tellurite emulsion of egg yolk recommended for identification of Staphylococcus species. Storage Between 2 to 8°C.

Specifications

Ingredient g/L Peptic digest of animal tissue 1.00 Sodium chloride 8.50 Final pH (at 25°C) : 7.0+ 0.2 Directions Suspend 9.5 grams in 1000 ml purified/distilled water. Heat if necessary, to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and dispense into tubes or flasks as desired. Principle The low concentration of peptic digest of animal tissue helps to maintain the organisms for 1-2 hours of dilution without multiplication. The isotonic property of the diluent ensures the recovery of organisms from various sources, which may be vulnerable in distilled water or aqueous suspensions. QC Tests (I) Dehydrated Medium Colour : White to pale yellow Appearance : Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating) : 7.0 ± 0.2 Colour (post autoclaving/heating) : Light yellow to colourless Clarity (post autoclaving/heating) : Clear solution without any precipitate (III) Q.C. Test Microbiological Cultural characteristics observed on Soyabean Casein Digest Agar , after an incubation at 35-37°C for 18-24 hours of cultures suspended in Maximum Recovery Diluent for 30 minutes. MICROORGANISM (ATCC ) RECOVERY (AFTER 30 MINUTES) Escherichia Coli (25922) no change in numbers Staphylococcus aureus (25923) no change in numbers Use It is a protective and isotonic medium used for maximal recovery of microorganisms from a variety of sources. Storage Dehydrated medium- below 30°C Prepared medium– Between 2 to 8°C

Specifications

Ingredient g/L Proteose peptone 10.00 Sodium chloride 5.00 Disodium phosphate 3.50 Monopotassium phosphate 1.50 Final pH (at 25°C) : 7.2 + 0.2 Directions Suspend 20.00 grams in 1000 ml distilled water. Heat if necessary, to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. If desired, aseptically add rehydrated contents of 1 vial of ECO157: H7 Selective Supplement for isolation of Escherichia coli O157 from foods to previously molten and cooled to 45-50°C medium. Mix well and dispense into sterile tubes or flasks as desired. Principle The media contain proteose peptone as a source of carbon, nitrogen, vitamins and minerals. Sodium chloride maintains the osmotic balance and phosphates buffer the medium. QC Tests (I) Dehydrated Medium Colour : Cream to Pale yellow Appearance : Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating) : 7.2 ± 0.2 Colour (post autoclaving/heating) : Light yellow to light amber Clarity (post autoclaving/heating) : Clear (III) Q.C. Test Microbiological Cultural characteristics observed after 18 –24 hrs at 35-37°C..(Recovery is carried out using XLD Agar,361) MICROORGANISM (ATCC ) GROWTH Salmonella enteritidis (13076 ) Good-luxuriant Salmonella typhi (6539 ) Good-luxuriant Salmonella typhimurium (14028 ) Good-luxuriant Escherichia coli 0157:H7 (NCTC 12900) Good-luxuriant (Recovery on tryptone soya broth) Use Used as pre -enrichment mediumincreasing the recovery of injured Salmonella species from foods prior to selective enrichment and isolation. Storage Dehydrated medium- below 30°C Prepared medium– Between 2 to 8°C

Specifications

Ingredients g/L Peptone 10.00 Sodium chloride 10.00 Final pH (at 25°C) : 8.4 + 0.2 Direction Suspend 20.0 grams in 1000 ml distilled water. Heat if necessary, to dissolve the medium completely. Dispense in tubes or flasks as desired. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Principle Peptone provides nitrogen and carbon source, long chain amino acids, vitamins and other essential nutrients. Sodium chloride maintains osmotic equilibrium QC Tests (I) Dehydrated Medium Colour : Cream to light yellow Appearance : Homogeneous Free Flowing powder (II) Rehydrated medium pH (post autoclaving/heating) : 8.4 ± 0.2 Colour (post autoclaving/heating) : Pale Yellow to light yellow Clarity (post autoclaving/heating) : Clear (III) Q.C. Test Microbiological Cultural characteristics observed after 18 –24 hrs at 35-37°C. MICROORGANISM (ATCC ) GROWTH Vibrio parahaemolyticus (17802 ) Luxuriant Vibrio cholerae (15748 ) Luxuriant Use For enrichment of Vibrio species. Storage Dehydrated medium-below30°C Prepared medium- Between 2 to 8°C.

Specifications

Ingredient Concentration (100 ml per vial) Egg yolk 30ml Sterile saline 70ml Direction Warm up the refrigerated egg yolk emulsion to room temperature. Shake well to attain uniform emulsion. (Since on refrigeration emulsion has a tendency to form layers or small lumps). Aseptically add 50 ml emulsion in 950 ml of sterile, molten, cooled (45-50°C) Baird Parker Agar Base, Mannitol Salt Agar Base, Bacillus Cereus Agar Base, Baird Parker Agar Base w/ Sulfate . or Aseptically add 100 ml emulsion in 900 ml of sterile, molten, cooled (45-50°C) Bacillus Cereus Agar base, McClung Toabe Agar Base, TPEY Agar Base, K.R.A.N.E.P. Agar Base, modified , MYP Agar Base (Phenol Red Egg Yolk Polymyxin Agar Base), Modified MYP Agar Base, KG Agar Base , L.D. Egg Yolk Agar Base / Egg Yolk Agar Base , Egg Yolk Agar Base, Modified or Aseptically add 25 ml emulsion in 475 ml of sterile, molten, cooled (45-50°C) Perfringens Agar Base T.S.C./S.F.P. Agar Base) , S.F.P. Agar Base , Bacillus Cereus Agar Base-Pemba .or Aseptically add 80 ml emulsion in 920 ml of sterile, molten, cooled (45-50°C) Anaerobic Egg Agar Base . or aseptically add 15 ml emulsion in 400 ml of sterile, molten, cooled (45-50°C) Willis and Hobb’s Medium Base . or aseptically add 7ml of Emulsion in 93ml of sterile, molten, cooled (45-50°C) Lipovitellin Salt Mannitol Agar Base . Mix well and pour into sterile petri plates. Use Sterile stabilized emulsion of egg yolk recommended for use in various culture media. Storage Between 2 to 8°C.

Specifications

Agar g/L 15.00 Beef extract g/L 8.00 Tryptone g/L 5.00 Peptone g/L 5.00 Sodium acetate g/L 5.00 Yeast extract g/L 1.00 Starch g/L 1.00 Dextrose g/L 1.00 Sodium bisulphite g/L 0.50 Ferric Ammonium Citrate g/L 0.50 L- Cysteine hydrochloride g/L 0.50 Resazurin mg/L 2.0

Specifications

Agar g/L 15.00 Casein enzymatic hydrolysate g/L 10.00 Sodium sulphite g/L 0.50 Ferric citrate g/L 0.50

Specifications

COMPOSITION Ingredients Gms / Ltr Peptic digest of animal tissue 10.000 Yeast extract 5.000 Sodium taurocholate 5.000 Sodium thiosulphate 10.000 Sodium chloride 20.000 Sodium lauryl sulphate 0.200 Sodium citrate 10.000 Sucrose 20.000 Bromo thymol blue 0.040 Thymol blue 0.040 Agar 20.000 Instruction for use :  Dissolve 100.28 grams in 1000 ml distilled water.  Heat to boiling to dissolve the medium completely, do not autoclave.  Mix well and pour into sterile Petri plates Principle The medium contains peptic digest of animal tissue and yeast extract, which provide nitrogenous compounds, vitamin B complex and other essential growth nutrients. Sucrose is added as a fermentable sugar. Sodium citrate, sodium lauryl sulphate, sodium taurocholate and sodium thiosulphate as well as high alkalinity of the medium inhibit most of the contaminating organisms. Bromothymol blue and thymol blue are the pH indicators. The alkaline pH of the medium and higher concentration of sodium chloride improves the recovery of Vibrio parahaemolyticus. Sucrose fermenting organisms like V. cholerae and V. alginolyticus produces yellow coloured colonies. Vibrio parahaemolyticus is a sucrose non-fermenting organism and produces blue-green colonies, as does V. vulnificus. Occasionally a few enteric sucrose nonfermenters may exhibit growth e.g. Proteus group.

Specifications

Ingredients gms/lit Soya Peptone 4.50 Sodium chloride 8.00 Potassium dihydrogen phosphate 0.60 Dipotassium phosphate 0.40 Magnesium chloride hexahydrate 29.00 Malachite green 0.036 Final pH (at 25°C) 5.2 + 0.2 Colour Pale greenish yellow