THE MICRO-ORGANISM OF SYPHILIS AND ALLIED ORGANISMS 211 



The India Ink Method. A drop of fluid from a lesion is 

 mixed with a drop of India ink upon a clean glass slide and 

 allowed to dry. Examine with oil-immersion lens. The 

 spirilla appear dark in a mass of carbon particles. By using 

 dark ground illumination, the organism appears brightly 

 refractive. 



Culture Methods. Noguchi, by using a serum water (i 

 part sheep or horse serum, 3 parts water, and adding a piece 

 of sterile rabbit's kidney or testicle), under strict anae- 

 robic conditions at 35 C. succeeded in cultivating the organ- 

 ism direct from lesions in man. After several transfers the 

 organism will grow on agar containing the bit of tissue. 



Inoculation Experiments. Pure cultures inoculated into 

 rabbits and monkeys produce lesions resembling .the primary 

 sores, and the blood of such animals gives a Wassermann 

 reaction. Cutaneous inoculation on eyebrows and genitals 

 of material from primary and secondary lesions produces 

 results in from fifteen to fifty days. 



Wassermann Reaction. In 1906 Wassermann, Neisser, 

 and Bruck described a method of making the diagnosis of 

 syphilis by demonstrating in the blood and spinal fluid of 

 a patient complement-binding substances not present in 

 normal blood. 



Technic. The following reagents are employed: (i) Syphi- 

 litic antigen; (2) serum to be tested; (3) fresh guinea-pig 

 serum; (4) washed sheep corpuscles and antisheep ambo- 

 ceptor. 



The antigen is an alcoholic extract of liver from a congenital 

 syphilitic, and is prepared by extracting the ground-up liver 

 with five volumes of absolute alcohol for ten days and then 

 filtering. 



Complement is normal guinea-pig serum. 



Antisheep amboceptor is obtained by injecting into a rabbit 

 2, 4, 6, 8, and 12 c.c. of washed sheep corpuscles on the first, 

 tenth, nineteenth, twenty-eighth, and thirty-seventh days 

 respectively. Nine days after the last injection the animal 

 is bled to death from the carotid and the blood collected in 



