370 BACTERIOLYSINS 



of each dilution, representing 2, 5, and 10 times the titer dose, are placed 

 in separate glasses or small test-tubes. A fourth tube should contain 2 

 c.c. of a 1 : 100 dilution of normal serum, according to the animal used 

 in producing the immune serum; a fifth tube should contain 2 c.c. of 

 sterile bouillon (culture control). 



Two loopfuls of suspected culture are thoroughly emulsified in each 

 of the five tubes of the series, and 1 c.c. of each is injected intraperi- 

 toneally in five guinea-pigs weighing about 250 grams each. 



At intervals of ten, twenty, forty, and sixty minutes the peritoneal 

 exudate should be removed with capillary pipets and examined in hang- 

 ing-drop preparations. Smears may be prepared and stained with dilute 

 carbolfuchsin, although they give less information than hanging-drop 

 preparations. 



If guinea-pigs Nos. 1, 2, and 3 show granule formation at the latest 

 after an hour, while in the fourth and fifth animals the bacteria remain 

 whole, motile, and well preserved, the reaction may be regarded as 

 positive and the diagnosis as established. 



Pfeiffer Bacteriolytic Test in vivo in the Diagnosis of Disease. Bac- 

 teriolysins are usually produced somewhat later than agglutinins, and 

 reach their highest point of production during convalescence. Bac- 

 teriolytic tests are used only for diagnostic purposes, when agglutination 

 reactions are negative or doubtful. The most satisfactory results from 

 these tests are obtained in cholera. Bacteriolysis with typhoid bacilli 

 is less typical and more incomplete; with the paratyphoid and dysen- 

 tery bacilli it is even more unsatisfactory, and in anthrax, pest, and the 

 various diseases due to cocci it does not occur. 



The test is conducted in a manner similar to the preceding test, ex- 

 cept that instead of an immune serum the patient's serum is used, with 

 a known culture of typhoid, cholera, paratyphoid, etc., according to the 

 infection suspected to be present. 



One cubic centimeter of the patient's serum is secured in a sterile 

 manner, inactivated by heating to 55 C. for one-half hour, and dilutions 

 of 1 : 20, 1 : 100, 1 : 250, and 1 : 500 prepared with sterile bouillon. Two 

 cubic centimeters of these dilutions are placed in separate tubes, and 

 two loopfuls of an eighteen-hour culture of the test organism emulsified 

 in each. A fifth tube contains 2 c.c. of bouillon with two loopfuls of 

 culture, and serves as the culture control. 



One cubic centimeter of each dilution and of the culture control is in- 

 jected intraperitoneally into five guinea-pigs, and the exudate examined 

 after twenty, forty, and sixty minutes. 



