CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



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solution, 2.5 cc. of a freshly prepared 

 sodium sulfite solution and 3.0 g. of 

 potato starch (Merck) stirred up in 

 5.0 cc. water, to (1) and steam in a 

 steamer for 30 minutes. 



(3) To a 50.0 cc. of beef serum diluted 

 with 5 parts water, add 0.5 cc. of a 

 10.0% NaOH solution and boil for 

 30 minutes in the steamer. 



(4) Mix two parts (2) with one part (1). 



(5) Distribute into sterile test tubes. 



(6) Pour sterile (5) into sterile Petri 

 dishes, seeing that the starch is well 

 distributed. Dry the plates before 

 use. 



Sterilization: Sterilize on three successive 

 days for 30 minutes each day in the 

 steamer. 



Use: Elective medium for cholera vibrio. 

 Author reported that after 3 to 9 hours 

 the cholera colonies were red. There 

 were two types, one a flat colony with 

 irregular edges and the other, smaller, 

 raised and definitely defined edge. Bad. 

 coli, Bad. dysenteriae Shiga-Kruse, 

 Bad. dysenteriae Flexner, anthrax bacil- 

 lus, Staphylococcus pyogenes aureus, 

 etc., all formed colorless colonies. The 

 cholera-like vibrio all gave one of the 

 two indicated types of red colonies. 

 Wheat starch gave the same results but 

 an hour later. Ordinary water soluble 

 starch (Merck) gave only the small well 

 defined edge type of colony. Medium 

 prepared with dextrin did not give as 

 good results as using potato starch. 



Variants: Kodama used 3.0 g. dextrin in- 

 stead of starch. 



Reference: Kodama (1922 p. 433). 



2008. Wasserman's Nutrose Serum Agar 



Constituents : 



1. Water 30.0 to 35.0 cc. 



2. Serum, hog 15.0 cc. 



3. Nutrose 0.8 to 0.9 g. 



4. Nutrient agar 



5. Peptone 



6. Glycerol 

 Preparation : 



(1) Place 15.0 cc. of hog serum as free 

 from hemoglobin as possible in an 

 Erlenmeyer flask and dilute with 30 

 to 35.0 cc. water. 



(2) Add 2 or 3.0 cc. of glycerol to (1). 



(3) Add 0.8 or 0.9 g. nutrose to (2). 



(4) Heat over a free flame to boiling, 

 shaking constantly. The turbid fluid 

 becomes clear. 



(5) Liquefy 2.0% peptone agar and cool 

 to about 50°C. 



(6) Mix equal parts (5) and sterile (4). 



(7) Pour into sterile Petri dishes. 

 Sterilization: Heat (4) until sterile. With 



fresh serum about 20 minutes is required. 

 With old serum a longer time is necessary 

 and it is often best to heat on several suc- 

 cessive days. Sterilization of agar not 

 given. 



Use: Cultivation of gonococci. 



Variants: Klimmer used alkaline nutrient 

 agar. 



References: Wasserman (1898 p. 300), 

 Klimmer (1923 p. 200). 



2009. Kligler's Nasal Secretion Serum Agar 



Constituents : 



1. Glucose agar. 



2. Serum, sheep. 



3. Nasal secretion. 

 Preparation : 



(1) Prepare glucose agar. 



(2) Obtain nasal secretions by blowing 

 the nose into sterile gauze. 



(3) Place into saline solution or alcohol 

 and keep 3 days. 



(4) Divide the saline mixture into two 

 equal parts. 



(5) Filter one of (4) thru a Berkefeld and 

 autoclave the other part. 



(6) Desiccate the alcoholic extract to 

 dryness and then take up in saline 

 solution. 



(7) Distribute (1) in 10.0 cc. lots. 



(8) Add 1.0 cc. sheep serum to each tube 

 of (7), also 1.0 cc. or 0.5 cc. of one of 

 the various extracts (6) and one of (5). 



(9) Pour into plates. 



Sterilization: Method of sterilization of 

 glucose agar not given. 



Use: Show influence of nasal secretion ex- 

 tract on growth of meningococci. Author 

 reported that saline extracts in each case 

 were superior to alcoholic extract. Fil- 

 tered saline extract more effective than 

 heated one. 



Reference: Kligler (1919 p. 39). 



