CULTURE MEDIA FOR CULTIVATION OF MICRO ORGAXISMS 



593 



3. Peptone 10.0 g. 



4. Agar 20.0 g. 



5. Glucose 10.0 g. 



6. K2HPO4 1.0 g. 



7. Blood, sterile defibrinated 



rabbit 100.0 cc. 



Preparation : 



(1) Chop 500.0 g. beef liver into small 

 pieces and boil for 2 hours in 1000.0 

 cc. of distilled water in a double 

 boiler. 



(2) Filter thru flannel and cotton. 



(3) Add to filtrate peptone and agar. 



(4) Heat in Arnold for one hour. 



(5) Adjust reaction as desired (for B. 

 bifidus, +1 to phenolphthalein). 



(6) Clear with eggs if necessary. 



(7) Add glucose and K2HPO4 to clear 

 filtrate. 



(8) To each 10.0 cc. of sterile medium 

 add about 1.0 cc. of sterile defibri- 

 nated rabbit blood just before pouring 

 the plate. 



Sterilization: Method not given. 



Use: Isolation of B. bifidus. To isolate 

 B. bifidus pour medium in petri dish. 

 Prepare nutrient agar seeded with B. 

 cereus and pour in cover of the Petri dish. 

 Allow the liver agar to solidify and 

 nutrient agar only partially. When 

 nutrient agar is semi solid, invert the 

 inoculated liver agar plate and place it 

 in the semi solid agar. The solidifying 

 agar forms a seal. This gives partial 

 anaerobiosis. Author reported colonies 

 were raised, more or less globular, opaque, 

 1 to 3 mm. in diameter, buff to reddish 

 brown in color. B. acidophilus gave 

 serrated edge colony — flat. To isolate 

 B. acidophilus use this medium omitting 

 blood and adjusting to +4 acid. Grow 

 aerobically. 



Variants: Harvey solidified medium 833 

 variant (a) by the addition of agar. 



References: Torrey (1922 p. 435), Harvey 

 (1921-22 p. 110). 



1883. Erickson and Albert's Testicular 

 Blood Agar 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Beef testicle 500.0 g. 



3. Peptone 20.0 g. 



4. Glucose 5.0 g. 



5. NaH2P04 2.0 to 3.0 g. 



6. Agar, granular 25.0 g. 



7. Blood, human 5.0 to 25.0 cc. 



Constituents : 



(1) Remove all connective tissue from 

 beef testicles, put thru meat grinder 

 and infuse on ice over night. 



(2) Next morning heat in a double boiler 

 to 50°C., allow to stand an hour and 

 then bring to the boiling point. 



(3) Let stand for another hour to permit 

 the solid particles to settle. 



(4) Decant off the liquid. 



(5) Dissolve 3, 4, 5 and 6 in (4) by heating 

 over a free flame stirring constantly. 



(6) Titrate using phenol red as an indi- 

 cator to pH = 7.4 to 7.8 or a reaction 

 of 0.6 to phenolphthalein. 



(7) Tube and autoclave at 15 pounds for 

 20 minutes. 



(8) Check titration. 



(9) While (8) is still liquid, add human 

 blood in proportion of 0.5 to 2.5%. 



Sterilization: See step (7) above. 



Use: Isolation and cultivation of gono- 

 cocci. Author reported that using phenol 

 red as an indicator primary acidity and 

 secondary alkalinity may be determined. 

 In a case of mixed infection, methyl violet 

 in proportion of 1:200,000 to 500,000 

 inhibited the growth of staphylococci. 



Variants: The authors substituted 1.0 to 

 5.0% defibrinated rabbit blood for human 

 blood. 



Reference: Erickson and Albert (1922 p. 

 277). 



1884. Harvey's Lactose Blood Agar 



Constituents: 



1. Infusion agar 1000.0 cc. 



2. Lactose 30.0 g. 



3. Blood, sterile defibrinated 

 human 



4. Rosolic acid (1.0%) 

 Preparation : 



(1) See variant (v) Medium 1661 for 

 preparation of infusion agar. 



(2) Mix 0.3 g. lactose with each 10.0 cc. 

 of (1). 



(3) Raise slowly to boiling water 

 temperature. 



(4) Cool to 45 °C. 



