SPIEOCHa:TOSIS OF SUDANESE FOWLS 81 



It was found that by this method living and fairly active spirochaetes could 

 sometimes be demonstrated thirty hours* after the putting up of the cover-glass 

 preparation, but in no instance, even when they entered corpuscles, did they coil up 

 or break down to form anything approaching the inclusions. These latter are clearly 

 visible by the dark-field method, appearing as silver rings or presenting appearances not 

 unlike tiny cartwheels, owing to their granular contents reflecting the light brilliantly, 

 while their vacuoloid areas remain dark. A "tucking in" of the corpuscular envelope at 

 the point where the body, if peripherally placed, was situated, was often apparent, 

 as, indeed, is the case in stained specimens. The method, however, indicated that the Breaking up 

 spirochaetes eventually break up into granules, and suggested the possibility that it is spirochetes 

 these granules which really invade the red cells. True, this has never been seen to into 

 happen in films of peripheral blood, but blood between a slide and cover-glass is not the S''*""^ 

 same as blood in blood-vessels, and, moreover, granting that these bodies really represent 

 a stage in the spirochaete life-cycle, it is quite likely that they enter the red cells in the 

 internal organs. At first this was merely a supposition. I killed chicks showing good 

 body infections and submitted emulsions of liver, spleen and lung and also the expressed 

 juice from these tissues to dark-field examination, finding a large number of free granules, 

 but it was not until I performed liver puncture and examined the blood thus obtained by the 

 dark-field method {vide page 104) that I obtained any definite evidence as to what really 

 seems to happen. Latterly, also, when employing "606" as a therapeutic agent, I have 

 obtained some very suggestive appearances in tissue stained by the Levaditi method {i-ide 

 pages 100 and 101). 



Acting on Dr. Stevenson's suggestion, I stained films of peripheral blood by the 

 Heidenhain method and counter-stained deeply with saffranin. The result went a long 

 way to show that the bodies have nothing to do with corpuscular or nuclear substance, 

 for, while the red cells stained a dull brownish-grey, their nuclei of course being of a darker 

 hue, the bodies stood out as brilliant pink dots (Plate III., fig. 8), taking on the saffranin 

 stain intensely, but unfortunately diifusely, so that no granular structure could be seen. 



It is true that the nuclei of the red cells, possibly because they contain chromatin, 

 exhibited, in some instances, pinkish granules, more especially when the corpuscles stained 

 faintly and were therefore probably degenerated, but these were unlike the bodies, being, 

 as a rule, darker and having irregular outlines. Thej' suggested nodes in the nuclear 

 network. Much the same aspect is presented by films stained by the Heidenhain method 

 and counter-stained with eosin. " Spore " forms of the bodies show black granules when 

 stained by the Levaditi-Yamamato method. 



When films were first of aU deheemoglobinised and then stained by either of these 

 two methods the bodies were found to retain the counter-stain for which they possessed an 

 afiBnity. No further light was, however, thrown on the problem. 



Other staining processes did not help one. The silver nitrate method of Khitrovo' Differential 

 for Sp. pallida was tried as was the Burri Indian ink method, but without any addition '' 



to our knowlege being gained. Wet fixation was adopted, but in vain so far as the 

 discovery of anything new as regards the morphology of the bodies was concerned. 

 In this respect one is not further advanced than at the time the Third Eeport was 

 written, though better specimens of the "spore" forms have been obtained which show 

 the inclusions forming or breaking up into granules and the discharge of the latter 



* In one instance I found a still-living spirochaete after 72 hours 

 ' Khitrovo (November, 1909), Archives Med. Beiges, quoted in Journal Royal Army Medical Corps, May, 1910. 



