REPORT OF THE BUERATJ OF ANIMAL INDUSTRY. 51 



this situation. For cover-glass preparations he found the following 

 solution to give the best results: Aniline water gentian violet,* or 

 fuchsin is mixed with an equal volume of .01 per cent, caustic potash, 

 or i per cent, liquor ammonia, directl y before using. The cover-g! ass 

 preparation, f is~stained by floating tb.e^ cover-glass with the film down 

 on the staining fluid for about five minutes. It is then dipped for a 

 second into 1 per cent, acetic acid to which a watery solution of tro- 

 paeoliii 00 t has been added until a wine-yellow color is obtained,, and 

 then washed in distilled water, after which operation it may be 

 examined or mounted in balsam for preservation. 



From the nodules and tubercles in the liver, spleen, and lungs the 

 contained pus was used to inoculate tubes containing various culture 

 media, both liquid and solid. A special value was put upon those 

 lesions which do not in any way come in contact with the external 

 air, such as those in the spleen and liver. Within three days the sur- 

 face of those tubes containing blood serum appeared as if sprinkled 

 with minute yellowish, translucent droplets. These droplets were 

 made up of very delicate rods or bacilli, varying in length between 

 one-third and two-thirds of a red corpuscle. Their width was from 

 one-fifth to one-eighth of their length. They were either straight 

 or slightly bent, with rounded extremities, and in general somewhat 

 shorter and thicker than tubercle bacilli. In liquids they manifested 

 active Brownian movement, but true spontaneous movements wore 

 never observed. They were without doubt identical with the deli- 

 cate rods observed in sections under the microscope. 



Of the biological character of bacteria, their growth or multiplica- 

 tion outside of the body on various substrata is very important, for 

 by this means of so-called cultivation we can determine how far they 

 are capable of growing in our environment and thus keeping up the 

 infection, between what temperature limits multiplication may take 

 place, also whether any resistant spore state is entered upon during 

 their life. There is still another advantage to be derived from cultiva- 

 tion one which mainly concerns us now the ability on our part to 

 diagnose between one organism and another by characters of growth 

 discernible with the unaided eye. The bacilli of glanders have cer- 

 tain features in cultures which are of great use in distinguishing 

 them from other organisms. 



They grow very well on blood serum from horses and sheep, less 

 abundantly on that from cattle. On the third day after inoculation 

 of such media, minute droplets of a translucent yellowish color ap- 

 pear on the surface of the serum. These droplets or "colonies" con- 

 sist entirely of bacilli which have descended from perhaps a single 

 bacillus originally deposited there. The material composing the 

 droplets is of a viscid consistency, and may be drawn out on the 

 platinum needle into thin threads. After eight or ten days this yel- 

 low translucency is replaced by a milky opacity. 



* This stain, used in the study of tubercle bacilli, is made by shaking up 5 cubic 

 centimeters of aniline oil in 100 cubic centimeters of water and filtering 1 . To the 

 filtrate is added 11 cubic centimeters of a concentrated alcoholic solution of the 

 aniline dye to be used, in this case either gentian violet or methyl violet or 

 fuchsin. 



f Prepared by crushing the nodule between the ends of forceps and rubbing it in 

 a very thin layer on the cover-glass. When thoroughly dry it is seized with the 

 forceps, drawn three times through the flame of a Bunsen burner, the film side 

 being held up or away from the flame. Pus from abscesses is rubbed in a very thin 

 film on cover-glasses and treated in the same way. 



\ This is not essential, 



