98 ROYAL SOCIETY OF CANADA 



Spores. — Frequent examinations of stained preparations from old 

 and young cultures failed to reveal the presence of spores. 



Stains. — The organism stained well with the ordinary aniline 

 colors and also by Gram's method. The bacilli stained with Loeflier's 

 alkaline-methylene-blue showed banded protoplasm or irregidar staining 

 with meta-chromic granules. 



Beef peptone IQfc gelatin was unfavorable for growth, and with 

 carbohydrates was unfavorable for pigment formation 6^ gelatin was 

 better, and washed gelatin best. 



Plate Cultures in Gelatin with Carbohydrates. — The medium used 

 in these experiments was made of: — gelatin 6^, peptone 1^, NaCl 

 0.5^, Liebig's b'eef extract, 0,25^, acidity -f 50. 



After the plates were poured and before the gelatin had set, th3 

 carbohydrates were added in the form of strong solutions in water. In 

 each plate both the bacteria and the carbohydrates were unequally dis- 

 tributed. The plates were incubated at 20° C. 



In 24 hours the colonies were minute and in 40 hours there was no 

 pigment. In 5 days the surface colonies were 1 m.m. in diameter, the 

 deep colonies 0.5 m.m. and liquefaction had begun in parts of the plate 

 where colonies were most numerous. The colonies were slimy and 

 could be drawn out in threads. The liquefied gelatin was not slimy as 

 long as the colonies in it remained discrete. 



Much pigment developed; the colors were of two kinds, blue and 

 violet in gelatin with sucrose, dextrose, mannite and maltose; and 

 green in gelatin with galactose, dextrin, starch, and with no carbohy- 

 drate. The violet was more the color of gentian violet than the 

 violaceus of Saccardo's chart. 



Pigment grains were numerous in the gelatin which developed a 

 strong violet and blue pigmentation. The colonies in maltose gelatin 

 were violet or violet centred and few pigment grains were present. 



The saccharose gelatin was bright blue in some parts and violet in 

 others, the dextrose gelatin and the mannite gelatin were generally 

 bright blue but were dark green in places. The colonies and the gelatin 

 were rich in pigment grains in those parts where the gelatin was colored, 

 i. e., where there was most sugar. 



The colonies in maltose gelatin were violet or violet-centred with 

 diffused pigment, a few pigment grains were present. 



The galactose gelatin was green and dark green and the colonies 

 the same color and rich in pigment grains or else brownish with none. 

 The dextrin gelatin showed less color and no grains in the colonies. 

 There was not sufficient dextrin added. The starch gelatin showed 

 scattered pigment grains. 



