Chemicals

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