INOCULATION EXPERIMENTS. 209 



acetic acid. Transfer for five minutes to fifty per cent, alcohol, 

 clarify in clove-oil, and mount in Canada balsam. 



Loffler*8 method: Sections are immersed for a few minutes in a 

 solution of potash 1 to 10,000, then for a few minutes in an alka- 

 line solution of methyl-blue, after which they are decolorized with 

 a solution of tropseolin in acetic acid, or, what is still better, in a 

 fluid composed often cubic centimetres of distilled water, two drops 

 of sulphuric acid, and one drop of a five per cent, solution of oxalic 

 acid. 



CULTIVATION. When cultivated on solid sterilized blood serum 

 at a temperature of 38 C. (100.4 F.), the growth appears in the 

 form of minute transparent drops, consisting entirely of the charac- 

 teristic bacilli. 



Potato cultures, according to Loffler, form in three days a uniform 

 amber-yellow layer which, about the sixth to the eighth day, assumes 

 a reddish hue, resembling the color of oxide of copper, which is not 

 easily mistaken for any other culture upon the same soil. Upon 

 this soil the bacilli were cultivated through twelve generations, and 

 the cultures retained their activity for a year; whether the bacillus 

 was capable of cultivation after this time is not mentioned. The 

 temperature at which cultures could be made to grow varied from 

 30 to 40 C. (86 to. 104 F.). 



Kranzfeld succeeded best with a nutrient medium composed of 

 meat-peptone, glycerin, agar-agar. The bacillus is destroyed by 

 exposure for ten minutes to a temperature of 55 C. (131 F.). A 

 three per cent, solution of carbolic acid, a one per cent, solution of 

 permanganate of potash, or a 1 : 5000 solution of corrosive subli- 

 mate destroys the bacilli with certainty. Lundgren succeeded in 

 obtaining an active culture in bouillon. 



INOCULATION EXPERIMENTS. Kitt ("Der Eauschbrand, Zu- 

 sammenfassende Skizze iiber den gegenwartigen Stand der Literatur 

 n lid Pathologic," Centralblatt fur Bacteriologie und Parasitenkunde, 

 Bd. i. S. 722, 1887) mentions the following animals susceptible of 

 inoculation with the virus of glanders : catttle, sheep, goats, guinea- 

 pigs. The horse, ass, and white rat are only susceptible to local 

 infection, the animals recovering completely and permanently after 

 a few days. Pigs, dogs, cats, rabbits, the common rat, ducks, and 

 chickens possess great immunity ; the inoculations at best produce 

 only a slight local reaction. Loffler made his first experiments on 

 guinea-pigs and the field mouse. In the guinea-pigs he observed, 

 three to five days after the subcutaneous injection of a pure culture, 

 an ulcer at the point of inoculation, and at the end of the first week 

 swelling of the nearest lymphatic glands in a state of purulent soft- 

 ening. At this stage of the disease the process often came to a 

 standstill and the animals recovered. In many animals the disease 



14 



