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CULTURE MEDIA FOR CULTIVATION OP MICROORGANISMS 



(2) Allow the blood to stand un- 

 disturbed for 15 to 30 minutes or 

 until the clot has firmly attached 

 itself to the sides of the vessel. 



(3) Cover the vessels and remove to 

 the laboratory. 



(4) Separate the clot from the sides of 

 the vessel by means of a sterile 

 knife, or a glass rod. Place the 

 vessel in the ice chest. 



(5) Allow to stand for 48 hours, and 

 pipette or siphon off the serum. 



(6) Place in tall cylinders (graduates) 

 and allow to stand 24 hours. 



(7) Separate the straw colored serum 

 from the sediment. 



(8) Add 0.5 cc. chloroform and pre- 

 serve in tightly stoppered flasks. 



(9) Mix one part of 1.0% glucose infu- 

 sion broth, with 3 parts (8). 



(10) Tube in sterile test tubes (about 3 

 cm. deep), and place in a sloping 

 position in an inspissator or 

 steamer and heat to 95°C. for 1 

 hour on 3 successive days, 

 (d) Heinemann (1905). 



(1) Collect fresh o.x blood in sterile 

 museum jars. 



(2) Set in an ice chest until the serum 

 has separated. 



(3) Filter the serum if necessary. 



(4) Take 3 parts (3) and mix thoroly 

 with 1 part of infusion broth 

 containing 1.5% glucose. 



(5) Tube. 



(6) Place in a Koch inspissator two or 

 three rows deep. Place about 

 25.0 cc. of water with the tubes. 



(7) Incline the inspissator to the 

 proper angle so as to slant the 

 serum. 



(8) Heat the water in the apparatus to 

 boiling and boil 5 minutes. Then 

 allow to cool. 



(9) Repeat (8) on the two following 

 days. 



(e) Rothe (1907). 



(1) Mix 4 parts sterile beef serum with 

 1 part sterile sugar free nutrient 

 bouillon (method of preparation 

 or composition not given). 



(2) Prepare 10.0% solution of levulose, 

 glucose, maltose, mannitol or 

 sucrose. 



(3) Sterilize (2) by heating at 100 °C. 

 either in a water bath or in a 

 steamer for 2 minutes on three 

 successive days. 



(4) To 90 parts (1) add 10 parts (3). 

 This makes the sugar concentra- 

 tion 1.0%. 



(5) Pour (4) into sterile petri dishes 

 m thin layers, and solidify. 



Roth reported that all diphtheria 

 bacilli fermented glucose and levu- 

 lose. Some pseudo diphtheria bacilli 

 fermented glucose and levulose. 

 (f) Abel (1912). 



(1) Dissolve 1.0% peptone, 0.5% NaCl 

 and 1.0% glucose in slightly 

 alkaline meat infusion. 



(2) Add 3 or 4 parts serum to 1 nart 

 (1). 



(3) Mix well and tube. 



(4) Solidify at a temperature between 

 90 and 95°C. 



(g) Drigalski and Bierast (1913). 



(1) Mix 600 cc. beef serum, 174 cc. 1% 

 dextrose bouillon, 26 cc. bile. 



(2) Pour into petri dishes in amounts 

 of about 16 cc. 



(3) Solidify at 90-95°C. 



(4) Sterilize fractionally on 3 succes- 

 sive days as with Loeffler serum. 



(h) Eastwood (1916) cultivated menin- 

 gococci on the following medium: 

 (1) Medium is composed of 3 parts 

 ox-blood serum, one part bouillon 

 with 1.0% of glucose, galactose, 

 malatose and sucrose respectively, 

 and 1 in 10,000 of neutral red is 

 added as an indicator. Details 

 of preparation not given, 

 (i) Shimer (1916). 



Add 1.4, 1.5 or 1.6 cc. of a 1.0% potas- 

 sium tellurate solution to Loeffler's 

 serum, 

 (j) Roddy (1917). 



(1) Preparation of glucose bouillon 

 not given. 



(2) Mix 1 part of (1) with 3 parts of 

 calf's or lamb's blood serum. 



(3) Tube. 



(4) Place in the serum sterilizer. 



(5) Heat at 57°C. for 1 hour each 

 day for 6 days and then at 70 °C. 1 

 hour each day for 2 days or heat in 



