672 
E. NOCARD. 
virulency, through continued generations upon rabbits, has 
been brought to its maximum of intensity and keeps it; and 
since 1891 we have always kept this condition of glanders to 
the same degree of virulency ; in using the pulp of the spleen 
of a rabbit that died 36 or 48 hours after an intra-venous in¬ 
jection, several tubes of potato cultures have been freely 
planted, and after three days of standing in a tempeiature of 
37 0 the product of a culture is diluted simply in a little quan¬ 
tity of boiled water, filtrated on linen and injected into the vein 
of the ear of a rabbit whose spleen will serve to the planting of 
other potatoes and so on. These potato cultures are not used 
for the preparation of the malleine but only to the cultivation 
of a glanderous virus with a high and always even virulency. 
To prepare malleine, we used cultures made on glycerined 
bouillon, according to the formula that we have given for the 
culture of the bacillus of tuberculosis. * 
Veal or horse bouillon, common salt, > 4 %, peptone, 1%, 
glycerine, 5%. 
Twenty-five to 30 bottles each of the capacity of 250 cubic 
centimeters and containing glycerined bouillon are planted at 
the same time, after a month of standing in the oven, the cul¬ 
tures are sterilized to the autoclave, then concentrated by 
evaporation in the water bath to the 10th of the first volum, 
then filtrated on Chiardin paper—thus is brute malleine obtained 
_that is a seripous liquid, of dark brown color, with aspeiral 
odor, somewhat vicous in aspect. Brute malleine containing 
50% of glycerine can be kept a long time, almost forever, away 
from the light and heat. In practice we have used it diluted to 
the 10th or to the 8th with phenic water at 0.5% 01 better in 
boiled water—but in this care the solution must be used as soon 
as it is prepared. 
1. In glanderous horses, the injection of the minimum quan¬ 
tity of malleine (one-fourth of one cubic centimeter of brute 
* No card's Rome.—^vx la culture du bacille de la tuberculose—Annales de 
l’lnstitut Pasteur, 25 Janvier, 1887. Page 27.) 
