156 MARTINI. 



Se\'en-day-ol(l enliurcs were chosen for prodiicing the infection, 

 Ix'CJiuse it was lioped that the iDirophasniata would liave died out in tlie 

 cuHures during this time, it already having l)een shown that the ti-ypano- 

 somata li\e longer than this. Nevertheless the results of this experiment 

 might liave heen accidental, since a piroplasmosis in this calf might have 

 existed and escaped observation. This point deserves especial considera- 

 tion, since Miyajima(7) (page 90) in performing similar experiments 

 observed on one occasion that in a calf inoculated with a trypanosoma 

 culture, after three days developed trypanosomata in its l)lood as was 

 proved by obtaining cultures of trypanosomata from it, while piroplasmata 

 were only found seven days later than this. 



Therefore, it was planned to repeat this experiment with piroplasmata 

 which had been subjected to conditions much more unfavorable to their 

 life and development. 



Calf No. 4 (re-petition of the experiments carried on vnth calf No. 2). — Feb- 

 ruary 26; new-born male calf from Indo-China; free from ticks; fed artificially; 

 blood free from parasites ; blood culture negative. A monkey was given 30 cubic 

 centimeters of blood subcutaneously and remained healthy and free from parasites. 



On the 2'6th of February calf No. 4 was inoculated with 30 cubic centimeters 

 of blood of the "original calf" subcutaneously. 



On the 5th of March piroplasmata, of the same appearance as those encoun- 

 tered in calves Nos. 1 and 2, were found present. 



On the 7th of March a bloody stool was passed. An examination of the 

 peripheral blood showed numerous red blood cells infected with piroplasmata. 



On the 10th of March a discharge from the eyes and nostrils appeared. Diar- 

 rhoea was present and the animal appeared to lose strength rapidly. 



On the 13th of March after the blood had been taken again for culture the 

 calf died. The cause of death was probably the same as in calves Nos. 1, 2, and 3. 



Autopsy. — Marked emaciation, discharge from eyes and nostrils; mesenteric 

 lymph glands hannorrhagic and swollen to the size of a pea or small bean; 

 catarrh of the large and small intestines; liver sligthly icteric. Otherwise no 

 pathological changes observed. The plasma bodies of Koch were not found in 

 smears either from the spleen or from the lymph glands. 



A blood culture nuide on the Sth of March showed no development of 

 protozoa, probal^ly on account of contamination with bacteria. 



Attempts at cultivation on the 9th, 10th, and 11th of March showed 

 the presence of forms which I had not encountered either in cultures from 

 the "original calf" or in tliose from calves Nos. 1 to 3. In these cultures 

 from calf No. 4 the piroplasmata showed no development into trypanoso- 

 mata, but on the other hand forms were found to be present which Eobert 

 Kocli(l,r') had encountered in ticks and described as the first stages of 

 development of Piroplasuia higeminum. (See Plate II, fig. 2, a, h, c, and 

 Plate VI, figs. 30 and 31.) 



On the first day of cultivation the piroplasmata, which showed two 

 distinct chromatin masses, were found to ha\e Ijecome free from the red 

 blood cells which they had severely injured. The injury to the red 



