CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 783 



the Arnold steam sterilizer at tion is complete. Do not heat 



80°C. 1 hour each day for 3 days. over 95 C. 



n^ rr. nQ1Q^ (6) Sterilize in the Arnold on each of 



(k) Tanner (1919). , , ,, . 3 successive days for 20 minutes. 



(1) Collect beef blood at the abatto. ^^^^^J^^^ZIZI (1887), Michel (1897 

 in clean --^--^ ^^ p^^^^, ^^ ^ p"261), Smith (1902 p. 104), Frost (1903 

 the ice box until clot is formed^ P Heineman (1905 p. 126), Rothe 



(2) Siphon or decant off the clear P^^^^ )> ^^^^^ ^^^^ ^^^ ^^^^^^.^ ^^^^^ 



'^®™™- , xi, / n 2471 Abel (1912 p. 25), Kolle Wasser- 



(3) Add one part glucose broth (prep- P; f '^f^^ U), Srigal'ski and Bierast 

 aration not given) to three parts rnan^i^ ^^p^^^ ^^^^J^^ ^^^^^ ^^ ^^^^ 



^^7"^- Shimer (1916 p. Ill), Roddy ^1917 p. 43), 



(4) Tube. , , . • ^ Tanner (1919 p. 69), Ball (1919, p. 80), 



(5) Coagulate (method not given). Tanner 919 P ; ^J ^^^^_^^ 



(6) Sterilize m the autoclave at about Besson ( P ^,^ ^ ,,^ (1921 

 112°C. or m the Arnold usmg the P- '^^\^J^ ^,,^, p. 367). Abbott 

 intermittent method if desired. P^ J39), ^^^^^ ^^^^^^^ ^^^^^ ^ ^^^^^ ^^.^^ 



(1) Harvey (1921-22). Klimmer (1923 p. 221, 222), 



Harvey added 2.0% or 3.0% chloro- a923 p^ 4 K J^^^^P.^^^ ^^^24 



form to the serum as a pre- -t^ark, vviiimui 



p. 124). 



servative. ^ 



(m) Park, Williams & Krumwiede (1924)^ ^^^^^^^ S^^^„ 



(1) Obtain beef or sheep blood from -^^^o- ■^'»" ^ 

 the slaughter house, and place constituents: 



in a sterile tall cylindrical vessel. ^ Bouillon lOO-^ ^^• 



(2) If the coagulum adheres to the 2' Serum, sheep 300.0 cc. 



sides of the vessel, loosen with a ^ Glucose 2.0 g. 



sterile glass rod. _ 4. Potassium-sulphocyanide. . . . 4.0 g. 



(3) Allow to stand for 24 hours on ice. ^' Neutral red (5.0% solution 



(4) Pipette or siphon off the serum. ' aqueous) 8.0 cc. 



(5) If bloody, place on ice and allow preparation: 



the corpuscle to settle out. ^^^ Method of preparation of bouillon 



(6) Mix 3 parts beef or sheep serum ^^^ gi^gn 



and 1 part nutrient broth (neutral ^2) Mix 3 parts good blood serum of the 



to litmus at a pH = 6.8 to 7.0) to ^^^^p ^^^ i part (1). 



which 1.0% dextrose has been (3) Add 3, 4 and 5 to (2). 



added. (4) Tube. 



(7) Tube. Sterilization: Coagulate and sterilize 



(8) Slant the tubes in an apparatus (^^^ct method not given). 



where the temperature can be j^-i^ ^f diphtheria. Author re- 



slowly raised to between 80 and u b aivhtheriae always pro- 



90°C. Do not heat above 95°C. Po^t^^ t ^^.^^ ^.^^^ ^^ 



until the medium is coagulated^ d^st nguished from cocci colonies. 



(9) Sterilize in the Arnold for 20 aisuufe 



minutes on each of 3 successive Vanants^:^^^^^^ ^^^^^ ^^^ ^^^^^^.^^ ^^^^^^ 



(n) ParkJNviUiams and Krumwiede ^^^^|-7::rp;sition and method of 



(ilTid 0.5% glucose to heart infusion preparation of nutrient broth not 



broth (see medium 794). (2)^0bTain fresh sheep blood serum 



(2) Mix 3 parts horse serum or plasma (2) Ubtam ire h 



.,, 1 „.t /n free from blood cells, 



with 1 part (1). ^^ ^^^ ^^^^^ ^^^ 25.0% (1), 0.5% 



^ If"^: glucose, 1.0% of 1.0% watery 



l5!H::tto80to90^C.untilcoagula- neutral red solution and 1.0% 



