BUEEAU OF ANIMAL INDUSTRY. 483 



ures were prepared by adding to each about four drops of the vaccine, 

 wliich, after exi)osure, had been kept in a jetVigerator. On the third 

 day one culture was i'aiutly clouded and found to contain the delicate 

 bacilli ; the other tube remained clear. The great majority of the bacilli 

 in the vaccine were evidently incapable of multiplyiug. That the cult- 

 ure liquid was not at fault was afterwards proved by inoculating the 

 tubes wliich had remained sterile with pure cultures of the lirst vaccine. 

 An abundant growth was observed in two days. 



(a) Inoculation of mice. — On October 28 two mice were inoculated, each 

 with one drop of the second vaccine, three with three drops each, and 

 one with four drops. The mouse inoculated with the tirst vaccine and 

 still well received about two drops. October 31 one of the mice inocu- 

 lated wiMi three drops was found dead. Liuibs partly Hexed, eyes half 

 closed. The organs were normal in appearance, excepting the lungs, 

 which were intensely congested. Cover-glass preparations from the 

 liver, spleen, blood from the heart and lungs contained no bacteria; a 

 meat broth pe]>tonc and a gelatine culture inoculated with blood from the 

 heart remained sterile. In this case death could not have been caused 

 by the inoculation. On ^November 4, one week after inoculation with 

 three droi)S uf the vaccine, a second mouse died, after having shown signs 

 of illness for a day or two pre%'ious. It was found with limbs flexed, as in 

 sitting, but it had fallen forwards on its head from a piece of bread on 

 which it had been sitting. The back was arched, the eyes closed with 

 slight secretion. On removing the skin of the abdomen a lew red patches 

 were found on its under surface. There was a collection of serum in the 

 fold of the groin on each side where the lymphatic glands stood out 

 prominent. Tliere was some serum in the peiitoneal cavity ; the spleen 

 was dark and tumelied, the liver quite dark, kidneys pale. On section 

 the cortex was paler than the medulla ; through both regions small rami- 

 fications of a dark red color extended ; lungs normal. Cover-glass prep- 

 arations revealed the minute bacilli in the blood from the heart, in the 

 lungs, spleen, liver, and kidneys. Their number in all organs, except- 

 ing in blood from the heart, was enormous. In the lungs, liver, and 

 spleen large pale cells were found in large numbers gorged with the 

 bacilli. (Plate II, Fig. 5.) Small cells were also observed in which the 

 stained bacilli were so numerous as to give the cell a dee])ly stained, 

 reticulated appearance. A gelatine and a liquid culture were prepared 

 by snip]iing away the apex of the heart with llamed scissors and trans- 

 ferring some blood with a platinum wire. These tubes, strange enough, 

 remained sterile. The remaining mii;e showed no signs of disease. 

 These experiments go far to demonstrate the identity of the pathogenic 

 bacilli in both vaccines. The large bacillus may have been some harm- 

 less microbe, the introduction of which into the culture took place no 

 doubt when the tubes were originally filled with the vaccine liquid. , 



AVe infer from the culture and inoculation experiments that the second 

 vaocine was as weak as the first, probably much weaker. The bacilli 

 of the first vaccine develo])ed abundantly on gelatine plates, while those 

 of the second did not. The latter failed in two out of three cases to 

 grow in liquid media, while the former readily grew in all inoculated 

 tubes. Their behavior towards the mice ag^rees with these determina- 

 tions. The efficiency of the vaccination, which depends on the quality 

 of the second as well as that of the first vaccine, is much impaired when 

 the strength and vitality of the preparation! cflnnok bo relied on. It is 

 true that there is much time lost in importiu j the vaccine from Paris 

 but as both vaccines were subjected to the same conditions, the fjreat 

 attenuation of the second vaccine in comiJarisou with the first must be 



