CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



657 



Preparation : 



(1) Clean and wash a number of pigs' 

 stomachs. 



(2) Mince (1) finely. 



(3) Add 10 cc. of strong hydrochloric 

 acid to 1000 cc. water at 50°C. 



(4) Add 200 g. (2) to (3) and keep at 

 50°C. for 20 hours. 



(5) Raise the temperature to the boiling 

 point. 



(6) Pour the mixture on a thick clean 

 cloth. Collect the fluid and that 

 obtained by squeezing the cloth and 

 its contents. 



(7) Heat the fluid to 80°C. and make 

 faintly alkaline to litmus while 

 at 80 °C. 



(8) Filter thru well-wetted thick filter 

 paper while hot. 



(9) Steam 30 minutes. 



(10) Filter again thru well-wetted thick 

 filter paper. 



(11) Mince finely, fat free veal, and add 

 500 g. to 1000 cc. water. 



(12) Incubate for 18 hours at 37°C. 



(13) Pour the mixture on a thick clean 

 cloth. 



(14) Collect the fluid, and that obtained 

 by squeezing the cloth, and its 

 contents. 



(15) Mix (14) with an equal volume of 

 pepsin digest solution (10). 



(16) Heat to 70°C. 



(17) Make the reaction neutral to litmus. 



(18) Add 7 cc. of normal NaOH per liter. 



(19) Filter thru well-wetted thick filter 

 paper. 



(20) Add sufficient agar to give a solid 

 medium. 



(21) Steam 30 minutes. 



(22) Filter while hot thru well-wetted 

 thick filter paper. 



(23) Add 1 cc. formalin to 500 cc. horse 

 serum. 



(24) Add sufficient 1% ammonia solu- 

 tion to neutralize (23) to litmus. 



(25) Mix one part (24) with two parts dis- 

 tilled water. 



(26) Mix one part sterile (25) with three 

 parts sterile (22). 



Sterilization: Method of sterilization of 

 (22) not specified. Sterilize (24) at 

 110°C. for 15 minutes in the autoclave. 



Use: Isolation and cultivation of meningo- 

 coccus. 

 Reference: Harvey (1921-22, p. 81). 



2042. Harvey's Defibrinated Blood Digest 

 Agar 



Constituents: 



1. Distilled Water. 



2. Veal, fat free. 



3. Minced stomach. 



4. HCl. 



5. Defibrinated blood. 



6. NaOH. 



7. Agar. 

 Preparation : 



(1) Clean and wash a number of pigs' 

 stomachs. 



(2) Mince (1) finely. 



(3) Add 10 cc. of strong hydrochloric 

 acid to 1000 cc. water at 50°C. 



(4) Add 200 g. (2) to (3) and keep at 

 50 °C. for 20 hours. 



(5) Raise the temperature to the boiling 

 point. 



(6) Pour the mixture on a thick clean 

 cloth. Collect the fluid and that 

 obtained by squeezing the cloth and 

 its contents. 



(7) Heat the fluid to 80°C. and make 

 faintly alkaline to litmus while at 

 80°C. 



(8) Filter thru well-wetted thick filter 

 paper while hot. 



(9) Steam 30 minutes. 



(10) Filter again thru well-wetted thick 

 filter paper. 



(11) Mince finely, fat free veal, and add 

 500 g. to 1000 cc. water. 



(12) Incubate for 18 hours at 37°C. 



(13) Pour the mixture on a thick clean 

 cloth. 



(14) Collect the fluid, and that obtained 

 by squeezing the cloth and its con- 

 tents. 



(15) Mix (14) with an equal volume of 

 pepsin digest solution (10). 



(16) Heat to 70°C. 



(17) Make the reaction neutral to litmus. 



(18) Add 7 cc. of normal NaOH per liter. 



(19) Filter thru well-wetted thick filter 

 paper. 



(20) Add sufficient agar to give a solid 

 medium. 



(21) Steam 30 minutes. 



