310 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Use: Cultivation and toxin production by 

 B. botulinus. Authors reported toxin 

 production very good in this medium. 



Reference: Dubovsky and Meyer (1922 

 p. 505). 



1002. Emery's Spleen Digest Solution 



Constituents : 



1. Water 1500.0 cc. 



2. Spleen (beef) 1500.0 g. 



3. Stomach (hog) 250.0 g. 



Preparation : 



(1) Digest 250.0 g. of beef spleen pulp, 

 250.0 g. hog stomach with 1.5 liters 

 of water acidulated with 15.0 cc. HCl 

 for 24 hours. 



(2) Add beef spleen pulp (500.0 g. to 1.0 

 liter of water) to (1). 



(3) Boil 10 minutes. 



(4) Filter thru wet absorbent cotton. 

 Sterilization: Not specified. 



Use: Differentiation of coli and typhoid 



bacilli. 

 Reference: Emery (1901 p. 979). 



1003. Stickel and Meyer's Blood Clot Digest 

 Solution 



Constituents : 



1. Tap water 1000.0 cc. 



2. Blood clots 100.0 g. 



3. Pig's stomach (minced) 100.0 g. 



4. K2HPO4 2.0 g. 



Preparation : 



(1) Obtain 10 liters fresh beef blood from 

 the abattoir. 



(2) Decant and store the serum (which 

 has separated on standing) in a 

 refrigerator. 



(3) Weigh the blood clots and mix 

 100.0 g. with 1 liter tap water. 



(4) Place mixture in an enameled pot, 

 bring slowly to a boil and boil slowly 

 for 5 minutes, stirring constantly. 



(5) Wash and mince pig's stomach. 



(6) Cool (4) to 50°C. and add 100.0 g. of 

 minced pig's stomach for each liter 

 of (4). 



(7) Transfer to glass or porcelain re- 

 ceptable and finally add 1.0% HCl. 



(8) Digest at 50°C. for 18-24 hours. 



(9) Make a Biuret and tryptophane, 

 test; when both are +, the digest is 

 yellowish green and contains very 

 little indigested debris. 



(10) Transfer to large bottles and steam 

 for 10 minutes to stop digestion. 



(11) Strain thru cotton or preferably 

 store over night in ice chest and 

 decant after 24 hours. 



(12) Warm (5) to 70°C. and neutralize 

 with 2 normal Na2C02 to litmus. 



(13) Filter into a flask. 



(14) Add the 2.0 g. K0HPO4. 



(15) Adjust to desired reaction using 

 litmus or preferably to a definite 

 H-ion concentration (pH = 7.0 

 to 7.5). 



(16) Clear (15) by adding 5-10.0% of the 

 decanted beef serum. Steam 45-60 

 minutes. 



(17) Remove (16) from steamer and 

 allow the clot to form as a compact 

 mass. Decant, or better centrifuge, 

 the medium to remove it. 



Sterilization: Sterilize at 100° for 30 



minutes on 2 successive days. 

 Use: General inexpensive culture medium. 

 Reference: Stickel and Meyer (1918 p. 81). 



1004. Bramigk's Peptic Digest Solution 



Constituents: 



1. Water 4000.0 cc. 



2. Blood clot 1000.0 g. 



3. Stomach (hog) 2 



Preparation : 



(1) Obtain a bucket of blood clots from 

 the slaughter house. 



(2) Wash the clots free from blood with 

 running water. 



(3) Press the clots free from water. 



(4) To 1000.0 g. clots (contains about 

 230.0 g. fibrin) add 3 liters of water 

 and 15.0 cc. of H2SO4. 



(5) Allow to stand over night. 



(6) Filter thru a sieve and press the 

 water from the clot or fibrin. 



(7) Add the clot to 3 liters water and 

 18.0 cc. H2SO4 heated to about 50°C. 



(8) Remove the mucous membrane from 

 two fresh pigs' stomachs. 



(9) Run the stomachs thru a meat chop- 

 ping machine. 



(10) Mix (9) with 1 liter of water and 

 8.0 cc. H2SO4. 



(11) Heat to 35°C. 



(12) Mix (11) and (7) thoroly and place 

 at 37°C. for 48 hours. Stir fre- 

 quently. Some fat will settle out. 



