CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 619 



(13) Autoclave for 15 minutes. (21) Pour large plates for use, with a 



(14) Add 150.0 cc. of Kahlbaum's lit- layer of medium about 4 mm. 

 mus solution and 15.0 g. lactose thick. . • ^u 



^gs (22) Place the plates so made, in the 



(15) Sterilize in the Arnold steamer for inverted position in the incubator 

 ^ ,^ . ^ without their covers and resting 



15 minutes. 



(16) Add 10.0 cc. of a hot 0.1% solution on a support. 



of crystal violet (B. Hochst). (h) Klimmer reported that tropon could 



„ be used instead of nutrose, that man- 



(l)"nnce finely fat free beef. nitol or maltose might be used in- 



2 Add 15.0 cc. to 2000.0 cc. water. stead of lactose, and a 1 0% Eich- 



3 Heat the mixture 20 minutes at a loff's solution may be substituted for 

 temperature not exceeding 50°C. meat infusion.^ (Eich off is a skmi 

 over a free flame. ^^^ preparation obtained from a 



(4) Skim off fat floating on the surface. food factory.) 



5 Raise the temperature to boiling References: v. Dngalski and Conradi 



^ ■ . (1902 p. 291), Frost (1903 p. 342), Heine- 



^fi^ Ron 10 minutes n^ann (1905 p. 130), Westbrook (1905 



? ur the mlTand fluid on to a p. 319), Reitz (1906 p^ 721), Abel (1912 



^^ wet, thick, clean cloth. PP- 128, 129) Ko^^^^^V TtTr^H v 



(8) Filter the fluid collected thru p. 415), Schmitz (1915 p- 307) Roddy 

 thick, well-wetted, filter paper. (1917 p. 44), Tanner (919 p^ 62) Ball 



(9) Add to the filtrate, peptone 20.0 g., (1919 p. 81), Gi U-er (1919 p 387), Be.an- 

 nutrose 20 g , sodium chloride ?on (1920 p. 342), Levine (1921 p. 116), 

 ?0 0g and see that the mixture is Abbott (1921 p. 523), Harvey (1921-22 

 alkaline to litmus. P- 90), Klimmer (1923 p. 208). 



(10) Steam 45 minutes. j^^y^ Galli-Valerio's Congo Red Nutrose 



(11) Add 60.0 g. prepared fiber agar * ^^^j. 

 (see medium 1401). 



(12) Steam gently 2§ hours or heat Constituents: onnn n „„ 



45 minutes at 118°C. to bring the 1. Water 2000.0 cc. 



agar thoroly into solution. 2. Meat, horse 1000.0 g. 



(13) Make faintly alkaline to litmus. 3. Peptone ^^-^ §• 



(14) Filter, while hot, thru well-wetted, 4. Nutrose ^U.u g. 



thick filter paper by placing filter 5. NaCl lU. g. 



funnel, stand and receptacle for 6. Agar W.u g. 



filtrate in the steam sterilizer and 7. Lactose -••••• ti'^ ^• 



steaming until filtration is com- 8. Congo red (1.0% soln.) 30.0 cc. 



plete. Preparation: 



(15) Boil 300.0 cc. litmus solution 10 (D Boil 1000.0 g. of horse meat with 

 minutes and dissolve in it 30.0 g. 2 liters of water for an hour, 

 chemically pure lactose. (2) Filter. ,, c^ ^ j u ;i 



(16) Boil the lactose litmus solution (3) Add 3, 4 and 5 to the filtrate and boil 

 15 minutes. ^ov an hour. 



(17) Mix hot lactose litmus solution and (4) Filter. , u * f 



the melted nutrose agar. (5) Add 60.0 g. of agar and heat for an 



(18) Make faintly alkaline if necessary hour m the autoclave 



by the addition of 10.0% sodium (6) Add soda solution unti the reaction 



hydroxide is slightly alkaline to litmus. 



(19) Add 4.0 cc. hot 10.0% sodium (7) Boil again for an hour and filter hot 

 hydroxide and 20.0 cc. hot sterile in the sterilizer , , , , , 

 1% c.p. crystal violet. (8) Distribute m 100.0 cc. lots and add 



(20) Distribute in flasks. 1-5 g- lactose and 30.0 cc. of 1.0% 



