308 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(9) Steam 60 minutes. 



(10) Pour the mixture on to a wet, thick, 

 clean cloth. 



(11) Collect the fluid which drains thru 

 the cloth together with that ob- 

 tained by squeezing the cloth. 



(12) Estimate and adjust the reaction of 

 the medium to a definite pH value or 

 to faintly alkaline to litmus or 1.0% 

 acid to phenolphthalein. 



(13) Filter thru well-wetted, thick, filter 

 paper. 



(14) Add 1 part of the filtrate to 2 parts 

 boiling water. 



Sterilization: Sterilize in the autoclave. 

 Use: General culture medium. 

 Reference: Harvey (1921-22 p. 69). 



997. Frouin and Ledebt'sHydrolyzed Serum 

 Solution 



Constituents: 



1. Water 1000.0 cc. 



2. NaCl 6.0 g 



3. KCl 0.3 g 



4. Potassium phosphate 0.5 g 



5. MgS04 0.3 g 



6. CaCla 0.15 g 



7. Serum albuminoids 10.0 g 



Preparation : 



(1) Dissolve 2, 3, 4, 5 and 6 in 1. 



(2) Hydrolyze 10.0 g. of serum albumi- 

 noids in (1). (Method not given.) 



(3) Neutralize (indicator not specified). 

 Sterilization: Method not given. 



Use: Cultivation of colon typhoid group. 

 Variants: The authors added 2.0% glucose 



or 3 to 4.0% glycerol to the medium. 

 Reference: Frouin and Ledebt (1911 p. 24). 



998. Martin's Stomach Digest Solution 

 (Martin's Peptone) 



Constituents : 



1. Water 1000.0 cc. 



2. Stomach (hog) 200.0 g. 



Preparation : 



(1) Chop the stomachs of five hogs into 

 small pieces. 



(2) Mix 200.0 g. of (1), 10.0 g. pure HCl 

 and 1000.0 cc. water at 50°C. 



(3) Allow to stand at 50°C. for 12 to 

 24 hours. 



(4) Heatatl0O°C. (Time not specified.) 



(5) Strain or filter thru a layer of ab- 

 sorbent cotton. 



(6) Heat the filtrate and make alkaline 

 when the temperature reaches about 

 80°C. 



(7) Filter the fluid thru paper to clarify. 



(8) Heat at 120°C. 



(9) Filter. 



(10) Distribute in flasks. 



Sterilization: Sterilize by heating at 115° 

 for 15 minutes. 



Use: Cultivation of diphtheria bacilli and 

 toxin production. The author reported 

 that the addition of 2.0 g. acetic acid per 

 liter favored toxin production. Other 

 organisms grew well on this medium. 



Variants : 



(a) Nicolle, Debains and Jouan culti- 

 vated meningococci on a medium 

 prepared as follows: 



(1) Heat one liter of water to about 

 55°C. 



(2) Add 10.0 cc. of pure HCl (22° 

 Baum's) to (1). 



(3) Add 300.0 g. of finely divided hog's 

 stomach. 



(4) Regulate the temperature to about 

 50 °C. and keep at this temperature 

 for 7 to 8 hours. 



(5) Heat to 80 or 90 degrees to destroy 

 the pepsin and stop digestion. 



(6) The peptone thus obtained may 

 be preserved for several weeks. 



(7) To use the peptone, make the 

 peptone slightly alkaline to litmus 

 and heat at 120 °C. 



(8) Filter on a wet filter paper. 



(9) Add 2.0 g. of glucose. 



(10) Sterilize at 112 to 115°C. 



(b) Besson used 250.0 g. hog's stomach 

 instead of 200.0 g. and digested for 

 20 to 24 hours instead of 12 to 24 

 hours. 



(c) Harvey digested for 20 hours and 

 steamed for 30 minutes instead of 

 heating at 120°C. as indicated in step 

 (8) above. 



(d) See medium 877, step (1) thru (8) 

 for Park, Williams and Krumwiede's 

 method of preparation. 



References: Martin (1898 p. 32), Thoinot 

 and Masselin (1902 p. 22), Nicolle, 

 Debains and Jouan (1918 p. 151), Besson 

 (1920 p. 28), Harvey (1921-22 p. 98), Park, 

 Williams and Krumwiede (1924 p. 118). 



