648 CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



ascitic fluid to (1) under aseptic Preparation: 



,^, ^ conditions. (1) Add 5.0 cc. of normal NaOH to a liter 



(b) Besson prepared the medium as of neutral 3.0% agar 



n l^^AxT: K f . •, • ^^^ "^^^ ^^^-^ ''• «^ ^ 10-0% nutrose solu- 



(1) Melt tubes of sterile nutrient agar. tion to sterile (1) 



(2) Cool to 45 to 50°C. (3) When ready for use liquify the agar 



(3) Add one-half or one-third the and cool to 45°C. 



volume of sterile ascitic fluid to (4) To each 100.0 cc. of (4) add 3 cc of 



M^ ^r^!f'-, u n- '^""^^ fil^^^^d ^^ bile, 1.0 cc. of a 



(4) Mix thoroly by rolling the tube 0.2% aqueous "Pure Safranin" (Grii- 

 between the palms of the hands. bier), 3.0 cc. of a 1.0% aqueous "Pure 



5) Slant or pour into a Petri dish. blue double concentrated" (Hoch- 



(c) Dopter and Sacquepee prepared the ster Farbwerke), and 3 to 4 cc 



nT7^TnT!''^!T"'^'- . •, °^ " ^-^^^ ^^^^«^« ^^l^^^ite green 



(1) Add 0.5 to 1.0 cc. of sterile ascitic crystals (Hochst.). Mix thoroly 

 fluid to agar slants. (5) Pour in plates. 



(2) Incline so that the entire surface Sterilization: Method of sterilization of 

 is covered with the serum. agar not given. Sterilize the bile by 



(3) Allow to stand for 12 to 24 hours. boiling 



References: Vellion (1898 p. 24), Thoinot Use: Enrichment of typhoid group. Klim- 



and Masselin (1902 p. 36), Besson (1920 mer reported that colon colonies were 



p 53), Bezangon (1920 p. 119), Dopter inhibited. Paratyphoid colonies grew 



n^^noo 2y?l^^^^^^ P- ^^^^' Klimmer luxuriantly as glassy, milk-like colonies 



(1923 p. 226), Stitt (1923 p. 42). References: Abel (1912 p. 132), KliLe; 



(1923 p. 214). 



2020. Herrold's Phosphate Ascitic Fluid 



Agar 2022. Hasting's Milk Agar 



Constituents: Constituents: 



1. Nutrient agar 300.0 cc. ^- ^'"trient agar 1000.0 cc. 



2. Na2HP04 ' ' 2. xAIilk 100.0 to 120.0 cc. 



3. Ascitic fluid 100.0 cc. Preparation: 



Preparation: (1) Prepare nutrient agar. 



(1) Prepare nutrient agar substituting ^2) Melt (1) and cool to 50°C. 

 Na2HP04 for NaCl. (3) Add 10.0 to 12.0% skimmed milk 



(2) Heat ascitic fluid at 56°C. for one *° (2). 



hour. (4) Tube and slant or pour in plates. 



(3) Add one part heated ascitic fluid to Sterilization: Not specified. 



three parts melted (1). Use: To demonstrate proteolysis. Hast- 



(4) Tube or pour into plates. i°gs reported that if proteolysis takes 

 Sterilization: Not specified. place the opaque medium clears. More 

 Use: Cultivation of streptococci for agglu- advantageous than gelatin to determine 



tination and absorption studies. proteolysis for milk agar may be incu- 



Reference: Herrold (1922 p. 80). bated at a temperature greater than 



20°C. Used also to cultivate fowl diph- 



2021. LoefHer's Dye Bile Agar (Abel) theria bacilli and B. vulgaricus. 



Variants : 



Constituents : (^^ .^j^U^^ reported the colonies of fowl 



1. Nurient agar (3.0%) 1000.0 cc. diphtheria bacilli appeared after 



Z- ^."Jt^ose 10.0 g. 40 hours when inoculated on the fol- 



/ ^'[^■■■. 20-0 ««• lowing medium: 



4. Safranin (0.2% soln.) 10.0 cc. (1) prepare nutrient agar. 



5. Pure blue (1.0% (2) Melt (1) and mix equal parts of 



„ Jr. uv 30.0 cc. melted agar and skim milk at 



0. Malachite green 100 °C 



^°-2^° "^^"■) 30.0 to 40.0 cc. (3) Pour into sterile tubes or plates. 



