128 The Philippine Journal of Science 1918 



inside the tissue cells, where the disinfectants cannot penetrate, 

 the virus in the blood stream being merely a surplus that is 

 thrown off from these tissue cells. In following this line of 

 reasoning, it was decided to consider certain tissues, where 

 lesions were more or less pronounced, as cultures, and extracts 

 were made from them. 



The tissues used in the following experiments were liver, 

 spleen, lymph glands, heart, intestines, thymus, skeletal muscle, 

 larynx, pharynx, and the back of the tongue from animals that 

 were either bled to death for virulent blood or that had died after 

 a regular course of the disease. 



The tissues were taken from the animal as soon after death 

 as possible. The amount of tissue desired was weighed and 

 then ground in a meat grinder that had been previously steril- 

 ized in the autoclave to keep external contamination to the 

 minimum. The material thus prepared was placed in a steril- 

 ized flask, and twice as much phenol solution (the strength of 

 which will be mentioned in each experiment) was added. Both 

 crude phenol and the pure crystal form were used in these ex- 

 periments with similar results, that is, 100 grams of liver were 

 ground and placed in a sterile flask, and 200 cubic centimeters 

 of a 0.5 per cent phenol solution were added to it. This material 

 was kept in the refrigerator, which averages between 15° and 

 16° C, and was daily thoroughly agitated two or three times. 

 In some experiments the material was placed in a shaking 

 machine and agitated continuously for forty-eight hours at room 

 temperature, which averages 26° C. in the morning and 28° 

 C. in the afternoon, some days rising to 30° C. After forty- 

 eight hours of agitation at room temperature the material was 

 placed in the refrigerator for twenty-four hours and then filtered 

 through gauze to separate the coarse material, and the filtrate 

 was replaced in the refrigerator until used. 



When the intestines were to be extracted, they were first 

 thoroughly washed free from faecal matter, then placed in a 0.5 

 per cent phenol solution for from five to ten minutes, after which 

 they were placed in a large container of boiled water, which 

 was cooled to at least 37° C. The tissue was allowed to 

 soak in this water for a few minutes to dilute the phenol that 

 remained intact. By this method a greater percentage of the 

 bacteria on the surface of the intestinal mucosa was destroyed. 

 Following this treatment the tissue was weighed, passed through 

 the meat grinder, and treated in a manner similar to that of the 

 other tissues. 



The animals used in these experiments were all highly sus- 



