380 MRS. L. J. TELET — CONTRIBUTION TO THE 



1896) confirmed these conclusions, as will be explained in the 

 section of this paper dealing more particularly with the refringent 

 bodies. 



Meanwhile, with regard to the bacteria, it was by working on 

 these lines that it was eventually found possible to follow the 

 development through all its stages, though it was not until 1899 

 that success was fully attained. 



In the spring of 1896 Mr. Bolton was again applied to for a 

 supply of PeloviyxcB, and 28 individuals were obtained. These 

 were chiefly used for work on the refringent bodies, but on 

 March 13 some rough attempts at cultivation of the bacteria 

 were made, using as a medium Mann's twice-filtered dilute 

 solution of egg-albumin in sterilized distilled water. The albumin, 

 being drawn straight from the egg, was tolerably germ-free, and 

 was put into a sterilized tube, to which a Pelomyxa, teased up 

 with a sterilized needle, was added, and the tube closed with 

 sterilized cotton-wool, and placed in the same temperature as 

 the vessel containing the supply of living Pelomyxw. 



On March 16 the tube was removed to a slightly warmer tem- 

 perature. In a few days a white filamentous growth was seen to 

 be proceeding from one of the teased-up portions of Pelomyxa 

 in the tube, and on microscopic examination was found to consist 

 of a pseudo-branching system, attached to one of the refringent 

 bodies, containing rods in a single-walled sheath, which broke 

 down, while under examination, to form free bacilli. 



A preparation was made of this, but, owing to the nature of 

 the medium in which the growth was formed, the preparation 

 showed too much stained deposit around the filaments and rods 

 to be instructive. At the time, also, not much importance was 

 attached to this development, owing to the fact of the albumin 

 not having been sterilized ; the growth was regarded as probably 

 belonging to the common species of Cladothrix (C. dicJiotoma), 

 which might have been ingested by the Pelomyxa, and con- 

 sequently as vitiating the experiment. It was not realized until 

 later that the actual development sought for had probably 

 been accidentally obtained. 



Further experiments at this time gave negative or mixed 

 results, and were beset with the diflBculties, formerly experienced 

 by Mr. Hill, of properly sterilizing the animal itself, as well as 

 the impossibility of satisfactorily sterilizing by heat a coagulable 

 medium 



