426 
Degeneration of T. gamhiense 
a solution of the stain in saline, in the manner described by Policard 
(1910). The tropho-nucleus was only slightly coloured by this stain 
but the kineto-nucleus and granules were stained a brick-red colour, 
and, in most cases, the latter were arranged in either one or two 
longitudinal rows in the region of the body between the tropho-nucleus 
and the anterior extremity. In one or two cases the extrusion of a 
granule from the tropho-nucleus was observed. Because of the fact 
that these granules may be found in some trypanosomes even at an 
early stage of infection, Policard infers that they are not degeneration 
products; but from the fact that the percentage of trypanosomes 
containing them goes up from 5 "/o—10 7o> before treatment, to almost 
100 7o after, when the parasites are certainly degenerating, it seems 
more reasonable to suppose that individual forms are dying off at all 
stages of infection and these contain granules. 
Up to the present it has been found impossible to demonstrate the 
axial filament by the use of intra vitam stains. 
Observations on the stained parasites. 
Of the methods employed wet films stained with iron haematoxylin, 
and dry films stained either with Azur or Giemsa, were found to be 
the most useful. The first method stains both nuclei together with 
the karyosome and intra-nuclear granules, also the degenerating axial 
filament (figs. 13—14) and certain chromatic granules occurring between 
the two nuclei (fig. 9). The granules occurring in that part of the cell 
anterior to the tropho-nucleus are not stained by this method, and 
therefore it is possible to distinguish two kinds, for both varieties stain 
equally well with Giemsa or Azur, which, in addition, colour all the 
other structures enumerated above. 
After staining with Giemsa both the intra- and extra-nuclear 
granules appear red, and in order to distinguish between them the films 
were afterwards treated with Gram’s Iodine Solution (Swellengrebel, 
1909). The films were first stained for ^ hr. with Giemsa, then rinsed 
with water and treated with Gram’s Iodine Solution for 1 min. and 
again washed in water. After this treatment the extra-nuclear granules 
of volutinose appeared black, sometimes with a reddish tinge; some of 
the granules within the tropho-nucleus appeared the same colour; the 
chromatin granules of the tropho-nucleus were greyish-brown; the 
kineto-nucleus appeared dark-red, and the plasma greyish-blue. 
