480 KEPORT OF THE COMMISSIONEE OF AGRICULTURE. 



TVlien stained in molhyl-violet, prepared by adding a drop of an alco- 

 holic solution to a watch glass of distilled water, and examined with a j\ 

 homogeneous objective, the. microbes appeared as bacilli in the form of 

 very slender lilameuts which at^sumed various curves, looi^s, and broken 

 lines. As the manner in which the vaccine had been x)ut up did not 

 seem to guarantee the absolute purity of the culture, a number of plate- 

 cultures vrerc made whence to obtain pure cultures. Two days later a 

 (gw scattered colonies appeared in the form of a rosette of club-shaped 

 elem?.nts; these were small bacteria. On the following day a large 

 number of very small round colonies could be discerned with a magni- 

 fication of about CO diameters. These were no doubt the fins bacilli. A 

 greenish film began to encroach upon the plates and liquefy the gela- 

 tine. A microbe was described by Schlitz as being in the vaccine used 

 in Baden which produced the same greenish coloration. Five days af^er 

 the plates had been prepared the colonies could not yet be distinguished 

 with the naked eye. Under a 1-inch objective they appeared as large 

 as pin's heads. To avoid total loss of the i)lates by the encroachment 

 of the chromogenous, putrefactive microbe, a spot was selected under 

 a dissecting microscope free from colonies of what were supposed to be 

 contaminating bacteria ; a minute portion of the gelatiuq layer contain- 

 ing a number of barely visible colonies was dug up and transferred to ' 

 10'="' of beef broth with 1 per cent, peptone. On the 23d, two days after 

 inoculation, a faint opalescence was observed in both tubes, which; on 

 shaking, was resolved for the moment into delicate rolling clouds. There 

 was no membrane or deposit. From another plate, a tube culture in nu- 

 tritive gelatine and one in beef broth with ijcptone were made in the 

 same way. In three days the liquid culture presented the same appear- 

 ance as those made from the other plate. Films, dried on cover-glasses, 

 and stained for half a minute, contained the same bacilli as those found 

 originally in Pasteur's liquid vaccine. The track of the platinum wire 

 in the tube of gelatine became opaque in a few days. A row of cloud- 

 like masses began to S]iread from it as a center and appeared as if strung 

 on the needle track which was now very faintly discernahle. (Plate II, 

 Fig. G.) These clouds were fringed at the edges, and when approaching 

 one another the entire growth i:)resented the ai:)pearance of a minute 

 test-tube brush* forced down into the trausi)areut gelatine. We were, 

 therefore, led to conclude from microscopical and culture appearances 

 that the microbe of Pasteur's vaccine was not a figure-of-eight form as 

 he himself described, but a bacillus not to be mistaken for a micrococcus. 



(a) Inoculation iciththejirst vaccine, in mice. — It had been experiment- 

 ally determined by Loeliier that the bacillus of rouget resembles very 

 closely the bacillus which produces septicaemia in mice, in microscopic 

 appearances, in its mode of growth in gelatine as well as in its behavior 

 towards the white blood corpuscles in the body of the Infected animal. 

 If the bacillus of Pasteur's vaccine is identical with the bacillus of 

 rouget in Germany, and not too attenuated, it should produce sei)tica}- 

 mia in mice. On October 17, three mice were inoculated by injecting 

 beneath the skin of the back of two 5 drops, of the third 10 drops of the 

 vaccine; they were kept in a large glass bell-jar covered with a sheet 

 of tin perforated with large holes over its entire surface, and suj^plied 

 abundantly with food and water. To avoid pain they were invariably 

 chloroformed before inoculation. October 20, one mouse was plainly ill ; 

 it moved with difficulty, had a staring coat and sulfused, partly closed, 



* Within a few days a book on rouget (Lydtin u. Schottelius Der EothJaufd. Schweine) 

 was recei\ed in whicli the term "test-tubo brush " is also used in describing the ap- 

 peararce of the gelatine culture. 



