558 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(5) Filter. 



(6) Adjust to pH = 7.1. 



(7) Add 10.0 cc. of Andrade's indicator. 



(8) Autoclave at 107°C. for 20 minutes. 



(9) Dissolve 5, 6 and 7 in the minimum 

 quantity of distilled water (about 

 30 cc). 



(10) Mix sterile (9) with (8) and stir with 

 a sterile glass rod. 



(11) Distribute into small flasks or tubes. 



(12) Place in the steamer for 10 minutes. 

 pH at this point should be near pH 

 = 7.4. 



(13) Prepare a 1.0% neutral lead acetate 

 (lead diacetate) by rapidly raising 

 30.0 cc. of sterile distilled water to a 

 boil, and adding 0.3 g. of neutral 

 lead acetate (lead diacetate). 



(14) Add (13) to (11) using a sterile 

 pipette in the proportion of 5.0 cc. 

 of (13) to each 100.0 cc. of (11). 



(15) Shake the flask quickly and 

 vigorously. 



(16) Add with a sterile pipette, 5,0 cc. of a 

 1.0% solution, Na2HP04 for each 

 100.0 cc. of medium. 



(17) Shake again. 



(18) Place in the water bath at 55°C. 



(19) Distribute into tubes. 

 Sterilization : Sterilize (9) by placing in the 



steamer for 20 minutes. See step (8) for 

 sterilization of agar. Final sterilization 

 of tubed medium not specified. 



Use: Enumeration of B. aertryke in feces. 

 The authors reported that 1.0% glucose 

 gave better results than lactose, sucrose 

 and salicin, if B. aertryke outnumber the 

 other organisms in the feces. 



Reference: Topley and Ayrton (1923-24 p. 

 230). 



1785. Frost's Glucose Agar 

 Constituents: 



1. Nutrient agar 1000.0 cc. 



2. Glucose (1.0%) 10.0 g. 



Preparation : 



(1) Add 1.0% glucose to nutrient agar. 



(2) Tube. 



Sterilization: Sterilize in the steamer. 



Use: General culture medium. 



Variants : 



(a) Heinemann added 1.5% glucose to 

 nutrient agar and sterilized the 

 medium in the autoclave at 120 'C. for 

 5 minutes. 



(b) Lohnis added 0.5% glucose, dissolved 

 in a little water to sterile melted 

 nutrient agar. 



(c) Lignieres added 0.25% agar and 

 0.25% glucose to peptone bouillon 

 and sterilized at 120°C. for 15 

 minutes. , 



(d) Giltner added 1.0% glucose and 1.0% 

 CaCOs to hot agar, and sterilized 

 by the discontinuous method. 



(e) Harvey added sufficient sterilized 

 precipitated chalk to render glucose- 

 agar white and opaque. 



(f) Puder cultivated Rhabditis pellio, a 

 nematode, on a medium prepared as 

 follows: 



(1) Prepare bouillon. Reaction to be 

 alkaline. 



(2) Mix 10.0 cc. of (1) with 90.0 cc. 

 water. 



(3) Dissolve 1.5 g. of agar in (2). 



(4) Add 2.0 g. of glucose and 8 drops of 

 concentrated (strength not given) 

 alkaline to (3). 



(5) Sterilize on three successive days 

 for 30 minutes each day in a 

 steamer. 



(6) Pour into sterile Petri dishes. 

 References: Frost (1903 p. 64), Heinemann 



(1905 p. 20), Lohnis (1913 p. 17), Lignieres 

 (1919 p. 1091), Giltner (1921 p. 365), 

 Harvey (1921-22 p. 89), Puder (1923 p. 

 99),Stitt(1923p.38). 



1786. Mankowaki's Indigo Carmine Glucose 

 Agar 



Constituents : 



1. 0.33 to 0.5% glucose agar. 



2. Fuchsin (acid, sat. solution in 1.0% 

 KOH). 



3. Indigo carmine (sat. aqueous solution). 

 Preparation : 



(1) Prepare a 0.33 to 0.5% glucose agar. 

 Reaction to be neutral. 



(2) Prepare a saturated solution of acid 

 fuchsin in a 1.0% KOH solution. 

 (Acid fuchsin may be added to a 1.0% 

 KOH solution until a dark black 

 brown color is reached.) 



(3) Prepare a watery saturated solution 

 of indigo carmine. 



(4) Add 2.0 cc. of (2) and 1 cc. of (3) 

 to 22 cc. of distilled water. This 

 solution is dark blue and reaction 

 slightly alkaline. 



