506 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(3) Allow to cool, skim and filter 

 thru filter paper. 



(4) Adjust the reaction so that the 

 acidity is equal to 0.025 g. H2SO4 

 per liter (0.5%). 



(5) Add 5.0 g. of agar and 10.0 g. De- 

 fresne peptone and heat for 15 min- 

 utes at 118°C. 



(6) Filter and tube in 6 to 8.0 cc. lots. 



(7) Heat at 112° for 15 minutes. 



(8) When ready for use add 0.1 cc. of a 

 1 to 10 sterile solution of lead sub- 

 acetate in distilled water to each 

 tube. 



(b) Tilley adjusted Jordan and Victor- 

 son's medium to pH = 7.2, sterilized 

 the agar in the autoclave and added 

 3 drops of freshly prepared 10.0% lead 

 acetate to each tube. 



(c) Harvey added 1.0 drop (0.05 cc. to 

 each tube (4.0 cc.) of agar (see 

 variant (v) medium 1661) cooled to 

 45°C. 



References: Jordan and Victorson (1917 

 p. 554), Emile-Weil (1917 p. 379), Tilley 

 (1923 p. 115), Harvey (1921-22 p. 107). 



1677. Elser and Huutoon's Basal Litmus 

 Infusion Agar 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Beef (lean) 500.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



5. Agar agar 16.0 g. 



6. Litmus, Kubel and Tie- 

 mann's 15.0 cc. 



Preparation : 



(1) Mix thoroughly 500.0 g. of chopped 

 lean beef with 1000.0 cc. distilled 

 water. 



(2) Boil over a free flame for 15 minutes, 

 stirring constantly. 



(3) Filter. 



(4) Add an emulsion of Bacterium coli. 



(5) Incubate at 37°C. for 24 hours. 



(6) Sterilize for 10 minutes. 



(7) Filter until clear. 



(8) Add 3, 4 and 5 that have previously 

 been soaked in water, to (7). 



(9) A portion of the filtrate is reinocu- 

 lated with Bacterium coli to estab- 

 lish the sugar free condition of the 

 product. 



(10) Place (8) in a saturated salt solution 

 bath and boil for 45 minutes, making 

 provision for loss of water by evap- 

 oration. 



(11) Add saturated Na2C03 solution until 

 slightly alkaline to litmus (allow for 

 a slight augmentation of the acid 

 during the subsequent steps). 



(12) Cool to 55°C. and add the white of 

 an egg and steam in Arnold sterilizer 

 until clarification of the upper 

 strata of the medium has occurred. 



(13) Pipette off the clear supernatant 

 fluid or filter. 



(14) Transfer definite quantities of (13) 

 to flasks. 



(15) To 100.0 cc. of hot sterilized distilled 

 water, add 10.0 g. of one of the added 

 nutrients, and expose the solution 

 to the action of live steam for 10 min- 

 utes. (Use old Jena glassware to 

 avoid alkaline production.) 



(16) Add to sterile (14) following the 

 third sterilization, 1.5% of sterilized 

 Kubel and Tiemann's litmus solu- 

 tion. 



(17) Transfer to sterile tubes, slant and 

 expose to incubator temperatures 

 for several days. 



(18) The final reaction of the medium 

 should be very faintly alkaline to 

 litmus. 



Sterilization: Sterilize (14) by steaming for 

 15 minutes on each of three successve 

 days. 

 Use: Cultivation of meningococci, pseudo- 

 meningococci and gonococci. The me- 

 dium is suited primarily for stock cul- 

 tures. 

 Added nutrients: The author added 1.0% 

 of one of the following: 



glucose mannitol 



galactose dulcitol 



levulose inulin 



lactose dextrin 



maltose sucrose 



Reference: Elser and Huntoon (1909 

 p. 406). 



1678. Emile-Weil's Neutral Red Infusion 

 Agar 



Constituents : 



1. Distilled water 1020.0 cc. 



2. Beef 500.0 g. 



