PRODUCTION OF BACTERIOLYSINS (fiACTERIOLYTIC SERUM) 69 



6. One week after the last injection has been made the blood is 

 tested, and if found of satisfactory titer, the animal is killed and the 

 serum secured. 



Intraperitoneal Method (Rabbit). 1. Same as the preceding, ex- 

 cepting that larger doses are given. 



First dose: 2 loopfuls in 4 c.c. NaCl solution, heated. 

 Second dose: 4 loopfuls in 4 c.c. NaCl solution, heated. 

 Third dose: 6 loopfuls in 4 c.c. NaCl solution, heated. 

 Fourth dose: 1 agar slant in 5 c.c. NaCl solution, heated. 

 Fifth dose: 1 agar slant in 5 c.c. NaCl solution, heated. 

 2. The blood is tested one week after the last injection has been 

 made. 



PRODUCTION OF IMMUNE OPSONINS 



1. These may be produced in the same manner as the agglutinating 

 serums, immune opsonins being readily demonstrated in the same se- 

 rums. For actual diagnostic work, artificial immune opsonins are sel- 

 dom required, but to secure an immune serum for experimental studies 

 on opsonins a culture of Staphylococcus pyogenes aureus may be used in 

 immunizing a guinea-pig as follows: 



First dose: 1 loopful of twenty-four-hour agar culture in 2 c.c. 



NaCl solution heated for one-half hour at 58C. and given 



subcutaneously. 

 Second dose: 1 loopful in 2 c.c. NaCl, heated; intraperi- 



toneally. 



Third dose: 2 loopfuls in 2 c.c. NaCl, heated; intraperitoneally. 

 Fourth dose: 3 loopfuls in 2 c.c. NaCl, heated; intraperitoneally. 

 Fifth dose: 6 loopfuls in 2 c.c. NaCl, heated; intraperitoneally. 



2. Bleed the animals one week after the last injection has been made. 



3. Owing to its large size, Bacillus anthracis may be substituted. 

 This is a spore-forming organism, and since it is dangerous unless scru- 

 pulous care in handling is exercised, it is not usually wise to employ it in 

 experimental work. 



PRODUCTION OF BACTERIOLYSINS (BACTERIOLYTIC SERUM) 

 1. These are prepared in exactly the same manner as agglutinins. 

 In practical diagnostic work the Spirillum cholerse is most frequently 

 used. In experimental studies of bacteriolysis the typhoid bacillus 

 and its immune serum may be employed with equal success. 



