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THE ROYAL SOCIETY OF CANADA 



As the wood ash medium and the Ashby medium seemed to be 

 far superior to the di-potass. phosphate a combination of both was 

 tried, equal parts of Ashby 's and wood ash media being mixed together. 



Growth X 



Moderate growth xx 



Spreading but thin xxx 



Thick but not spreading xxxx 

 Thick and spreading xxxxx 



Bottom row symbols — 6 days' growth . 

 Room temp. 65-70 F. 

 Top row of symbols — 14 days' growth, 

 Room temp. 65-70 F. 



Mannite gave the best results with the red clover variety; but 

 as a whole the saccharose gave the best growths. The addition of 

 mannite to the media seemed to give an early start to the organisms 

 and in subsequent lots of media a small quantity of mannite mixed 

 with saccharose was used with excellent results. 



For the last two years we have used 1 per cent saccharose and 

 i per cent mannite with good results. Growth starts quickly, becomes 

 thick and spreads rapidly to the sides of the bottle. The red clover 

 organism grows less fast, however, and does not form such a thick 

 layer. 



As a result of these experiments the medium used for our culture 

 is made as follows: Three solutions are prepared — 

 A. — 75 grams of agar are dissolved in 3,000 c.c. of water, by 



placing in the autoclav at 10-15 lbs. pressure. 

 B. — 25 grams of hard wood ashes are boiled in 1,000 c.c. of water and 



filtered. 

 C. — -5 gram of acid potassium phosphate, 

 •5 gram of magnesium sulphate, 

 •5 gram sodium chloride, 

 • 25 gram calcium sulphate, 

 6 • 25 grams of calcium carbonate 

 are dissolved in the order named in 1 ,000 c.c. of hot water (about 80°C.) 



The three solutions, A B and C are mixed and 87-5 grams 

 saccharose and 12-5 grams of mannite are added. The resulting 5 



