162 CLINICAL BACTERIOLOGY AND 



These cultures so derived are incubated in broth-tubes for 

 five consecutive days at a temperature of 42° C, and from 

 this Vaccine No. I. is derived. Vaccine No. II. is obtained 

 in the same way, except that incubation goes on for two 

 days only. The dose advised by Lignieres is 1 c.c. of 

 Vaccine No. I. to be injected into the thigh of a puppy; 

 ten days later Vaccine No. II. is injected into the opposite 

 thigh. 



Lignieres at a later date injected his vaccine as above 

 described into horses, beginning with small doses, and 

 increasing them as toleration became established at regular 

 intervals, first under the skin, and later into the vein 

 direct. 



The immune serum is then obtained by bleeding the 

 animal, collecting the blood in sterile vessels, allowing it 

 to clot on ice, and the resultant serum placed in stock 

 bottles, and a small percentage of lysol added. 



The dose is from 10 to 20 c.c. to each dog. 



Phisalix, adopting Lignieres' method, made his mono- 

 valent vaccine, which he derived also from a pasteurella 

 cultivated on glycerine and broth. The dose he gives is 

 2 c.c. to each dog, repeating the vaccine in ten days. 



Piorkowski prepares a polyvalent serum which is highly 

 spoken of by many clinicians, while others consider it is 

 useless as a prophylactic and curative agent. 



As a prophylactic dose he advises 5 to 10 c.c, im- 

 munity, it is stated, being maintained thereby for six 

 months, and for a curative dose 10 to 20 c.c. is recom- 

 mended. 



Ferry, after isolating his microbe, the B. bronchosepticus, 

 made a protective vaccine mainly to prove whether this 

 bacillus was the real causative organism of distemper, and, 

 in addition, he carried out agglutination tests. 



The following is an extract taken from his paper, " The 

 Cause of Distemper in Dogs," and which appeared in the 

 New York Journal for July, 1912 : 



"Agglutination Tests and Protective Inoculations. — In 



