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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(4) Sterilize 3 times for 1.2 hour each 

 time (exact method not given). 



(b) Thoinot and Masselin (1902). 



(1) Sterilize pieces of potato properly 

 prepared in Roux tubes. 



(2) Add 4.0% glycerol in distilled 

 water or bouillon to each tube to 

 fill the tube to the constriction in 

 the tube. 



(3) After inoculation slant the tubes 

 so that the glycerol is in contact 

 with the potato. 



(c) Smith (1902). 



(1) Wash large sound potatoes in 

 running water using a stiff brush. 



(2) Remove all the eyes and any suspi- 

 cious spots. 



(3) Cut the ends from the potatoes 

 and cut cylindrical pieces using a 

 cork borer. 



(4) Cut the cylinders obliquely so 

 that each resulting piece presents 

 a round flat end and a large oval 

 beveled surface. 



(5) Place the pieces in running water 

 for several hours or over night if 

 convenient. 



(6) Place one piece of potato into 

 sterile tubes containing a bit of 

 glass rod. The beveled side of 

 the potato should be up. 



(7) Add a 5.0% glycerol solution up to 

 the level of the lower surface of 

 the potato. 



(8) Sterilize by heating at 100 °C. for 

 30 minutes on one day and for 

 15 minutes on the second and third 

 successive days, or in the auto- 

 clave at 120°C. for 30 to 40 minutes. 



(d) Heinemann (1905). 



(1) Prepare potato slants in the ordi- 

 nary manner. 



(2) Soak the potato slants in 25.0% 

 solution of glycerol. 



(3) Add a little glycerol solution to 

 the bottom of the tube. 



(e) Anzilotti (1906). 



(1) Wash sound potatoes. 



(2) Cut cylindrical pieces from the 

 potatoes using a cork borer. 



(3) Split the cylinders, and wash the 

 potatoes carefully in distilled 

 water. 



(4) Boil the potatoes in a 6.0% glycerol 



solution that has been made alka- 

 line by the addition of concen- 

 trated solution of Na2C03. Boil 

 until the potatoes have become 

 soft and swollen usually requires 

 20 minutes. It is necessary to 

 keep the reaction alkaline by the 

 addition of more Na2C03 solution. 



(5) Place the pieces of potato into 

 sterile Roux tubes, or a piece of 

 glass tubing may be placed in an 

 ordinary tube so that the potato 

 will be about 2 mm. from the bot- 

 tom of the tube. 



(6) Pour the alkaline 6.0% glycerol 

 solution into the tubes so that the 

 bottom of the potato is covered 

 with the solution. 



(7) Plug the tubes with cotton. 



(8) Steam on 3 successive days for 

 20 minutes in a Koch apparatus, 

 or sterilize for 20 minutes in an 

 autoclave at 120°C. 



(f ) Peklo impregnated potato slants with 

 3.0% glycerol solution. 



(g) Park and Krumwiede (1910). 



(1) Cut potatoes in usual Bolton 

 method (Details of method not 

 specified). 



(2) Soak in 1-1000 sodium carbonate 

 solution in distilled water for 

 24 hours. 



(3) Drain. 



(4) Soak in 5.0% glycerol in distilled 

 water solution for 24 hours. 



(5) Tube, using | inch tubes and add 

 glycerol water in which they were 

 soaked to cover about \ of the 

 surface. In some instances, to 

 obtain a large number of pellicles, 

 5.0% glycerol bouillon was used 

 to cover the potato. Pellicle 

 formation good using plain glyc- 

 erol water, however. 



(6) Sterilize by the intermittent 

 method in the Arnold sterilizer. 



(7) After inoculation seal the tubes 

 either with cork alone or prefer- 

 ably dip the cotton stopper also 

 in paraffin. 



(h) Nocard (Roux and Rochaix) (1911). 

 (1) Soak pieces of peeled potato in 

 15.0% glycerol for 48 hours. 



