476 TRANSACTIONS OF THE CANADIAN INSTITUTE. [Vou. VI. 
tributed uniformly throughout the cytoplasm and that this distribution 
corresponds with the diffuse stain given by hamatoxylin. In the 
“comma” forms granules which stained with hematoxylin gave a re- 
action for “ masked” iron. 
MATERIAL AND METHODS OF STUDY. 
The forms used were Beggzatoa media, B. alba and B&. mirabilis. The 
cultures of the two former in water containing sulphuretted hydrogen in 
solution were always kept in the actively growing condition. These 
cultures could in twenty-four hours be got to yield myriads of the 
spirillum-like elements, the “comma” forms, and the “cocci,” which, 
according to Zopft are different stages in the development of 
Beggtatoa.’ 
One method of fixation was to place a drop of the culture on a cover 
glass, allow the water to evaporate, and then to float the cover, prepara- 
tion surface downwards, on a saturated solution of corrosive sublimate, 
where it was left for a couple of hours, after which it was passed succes- 
sively through alcohols of fifty, seventy and ninety per cent. strengths. 
Cover glass preparations were made with ninety-five per cent. alcohol, 
without the employment of any other reagent. Picric acid in saturated 
solution was also employed on cover preparations as well as on 
quantities of the material in various stages. Material in bulk was 
hardened in ninety-five per cent. alcohol alone. 
In staining, methylene-blue, safranin, eosin and Ehrlich’s and Heiden- 
hain’s hematoxylins were employed. The first three dyes, each used 
alone, give results of no value, but they may individually be employed 
with one of the hematoxylins and thereby a more marked demonstra- 
tion of the vesicular structure of the various forms may be obtained. 
If, however, the iron-alum hematoxylin method alone is carefully 
employed it will give preparations which in distinctness leave nothing 
to be desired. 
The material from B&B. mzrabelis was hardened, part in absolute. 
alcohol and part in picric acid in saturated aqueous solution.’ 
1 ‘‘Znr Morphologie der Spaltpflanzen,”’ Leipzig, 1882. Also ‘‘ Die Spaltpilze,’’ Breslau, 1883. 
z These forms are, according to Winogradsky (‘ Beitrage zur Morphologie und Physiologie der 
Bacterien,” Leipzig, 1888), not geneticallv related to Beggiatoa. He was unable to find a transformation 
of the Beggiatoa threads into the ‘‘cocci” forms, an experience which befell Engler (‘‘ Ueber die Pilz- 
Vegetation des weissen oder todten Grundes in der Kieler Bucht,’’ Vierter Bericht d, Commission zur Wiss. 
Untersuchung der deutschen Meere in Kiel, p. 185, Berlin, 1884). 
3 For this material Iam indebted through Dr. E. C. Jeffrey to Mr. Billings. 
