736 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(13) Filter thru absorbent cotton and 

 cotton flannel, passing the filtrate 

 thru the filter until clear. 



(14) Titrate and record the final 

 reaction. 



(15) Tube, using 5.0 cc. in each tube 

 in the case of gelatin. 



(16) Sterilize 15 minutes in the auto- 

 clave at 110° or for 30 minutes in 

 streaming steam on three successive 

 days. 



(17) Store in the ice chest in a moist 

 atmosphere to prevent evaporation. 



(b) Committee A. P. H. A. (1905) pre- 

 pared the medium as given in variant 

 (a), but sterilized for 5 minutes in 

 the autoclave at 120 °C. or for 30 

 minutes in streaming steam on each 

 of 3 successive days. Cool quickly 

 on ice following heating. 



(c) Giltner prepared the medium as 

 follows: 



(1) Prepare meat infusion. 



(2) Dissolve 1.0% peptone and 15.0% 

 gelatin in a mixture of equal parts 

 (1) and distilled water. 



(3) Adjust the reaction to +1.0%. 



(4) Sterilization not specified. 

 References: Smith (1897 p. 480), Com- 

 mittee A. P. H. A. (1901 p. 384), (1905 

 p. 107), Giltner (1921 p. 380). 



2282. LoefHer's Malachite Green Infusion 

 Gelatin 



Constituents : 



1. Water 5000.0 cc. 



2. Beef 4.0 lbs. 



3. Gelatin (15.0%) 750.0 g. 



4. Peptone (Witte 1.0%) 50.0 g. 



5. XaCl(0.5%) 25.0 g. 



6. Malachite green solution 



7. Phosphoric acid 

 Preparation : 



(1) Chop four pounds clean lean beef. 



(2) Mix 5 liters tap water with (1). 



(3) Add 15.0% = 750.0 g. gelatin, 1.0% 

 = 50.0 g. Witte peptone, 0.5% = 

 25.0 g. NaCl. 



(4) Slowly heat to complete solution of 

 gelatin. 



(5) Heat 45 minutes. 



(6) Neutralize to litmus with sodium 

 carbonate. 



(7) Heat and filter. 



(8) To each 100.0 cc. add 3.0 cc. of a 

 normal phosphoric acid and 2.0 cc. 

 of 2.0% malachite green solution. 

 Sterilization: Not specified. 

 Use: Isolation of typhoid bacilli from 



feces. 

 References: Loeffler (1906p. 289), Klimmer 

 (1923 p. 213). 



2283. Roux and Rochaix's Peptone Infusion 



Gelatin 

 Constituents : 



1. Distilled water 1000.0 cc. 



2. Beef 500.O g. 



3. NaCl 5.0 g. 



4. Peptone 10. g. 



5. Sodium or potassium 

 phosphate 2.0 g. 



6. Gelatin (15.0 to 



18-0%) 150.0 to 180.0 g. 



Preparation : 



(1) Chop 500.0 g. of fat and tendon free 

 beef into very small pieces. 



(2) Add 1 liter of water and allow to 

 stand over night in the cold. 



(3) Press the juice thru a linen cloth 

 by means of a meat press. 



(4) Make up the volume to 1000.0 cc. 

 by the addition of water. 



(5) Add 5.0 g. NaCl, 2.0 g. of sodium or 

 potassium phosphate and 10.0 g. of 

 peptone. 



(6) Boil for an hour. 



(7) Filter thru a linen cloth and then 

 thru paper. 



(8) Adjust the reaction slightly alkaline 

 to litmus by addition of KHCO3 

 solution, drop by drop. 



(9) Add 15.0 to 18.0% gelatin and heat 

 on the water bath until solution is 

 complete. 



(10) Neutralize and make slightly alka- 

 line again to litmus. 



(11) Filter using a hot water funnel. 



(12) Distribute in flasks or tubes. 

 Sterilization : Sterilize on 2 or 3 successive 



days at 100 °C. 

 Use: General culture medium. 

 Variants : 



(a) Choquet's (Besson) prepared a me- 

 dium as follows: 



(1) Dissolve 5.0 g. peptone and 35.0 g. 

 gelatin in 500.0 cc. beef juice. 



(2) Heat in the autoclave. 



