﻿450 ASHBURN AND CRAIG. 



containing scattered treponema lias as many organisms as another prep- 

 aration in which they are in clumps. Novy and Knapp, who describe 

 agglutination of Spirillum obermeieri say that in the case of that organ- 

 ism there is no increase in number. Counts being out of the question, 

 the observers' estimate must be relied upon. Our belief, based on many 

 examinations of sera made on the first, second, and third days, etc., 

 is that in most instances Treponema pertenuis multiplies greatly when 

 kept in the serum in capillary tubes, and in some instances the in- 

 crease seems to occur without the agglutination. Our opinion that 

 the organisms multiply in such preparations is based not only on the 

 greater number of them found in the tube preparations as compared with 

 smears made at the same time, but also on the appearance of the clumps 

 and the great preponderance of what we consider dividing forms as 

 described under types I, K and L. The nature of some such clumps 

 and dividing forms are shown in the accompanying photomicrographs. 

 (PI. 1, figs. 5 and 6.) 



Exceptionally, neither multiplication nor agglutination developed in 

 the tubes, and the failures, while infrequent, are somewhat irregular and 

 do not admit of what we consider a thoroughly satisfactory explanation. 

 However, in general terms, we are of the opinion that they are due to 

 one or both of the following reasons : 



First. Variability in the immunity of the yaws patient and in the 

 amount of antibodies contained in the serum ; possibly the variability of 

 immunity may even be local, or toxins elaborated by bacteria may be 

 present locally to inhibit the growth. An interesting observation which 

 may bear on this point was made in a case of syphilis. 



Serum from a chancre was taken up in tubes on Monday. The 

 chancre was then washed with bichloride solution and dressed with 

 calomel. On Tuesday it was thoroughly cleaned and more serum placed 

 in tubes. The tubes of Monday showed marked increase and clumping 

 of treponema, those of Tuesday, neither. The patient was not taking 

 any general treatment. 



Second. Variability in the bacterial content of the tubes and conse- 

 quently in their content of soluble toxins. 



While it is readily possible to make fresh smear preparations of yaws 

 serum which show very few or no bacteria, it has not been possible, in 

 our experience, to obtain a serum really free from these organisms, and 

 in the sealed tubes they multiply enormously. However, the fact that 

 both multiplication and clumping occur in the great majority of tubes 

 encouraged us in the idea that we might cultivate the treponema indefi- 

 nitely, and as the favorable medium seemed to be the serum of sus- 

 ceptible persons or animals, we endeavored to obtain growth on monkey 

 blood and on ascitic fluid from a patient who was suffering from cirrhosis 

 of the liver who also gave a history of syphilis. The ascitic fluid was 



