ANTIGENS AS BIOCATALYSTS 31 



non-specific substances, such as pilocarpin stimulated the restoration 

 of the antibody formation. One unit of tetanus toxin produced (Knorr, 

 1898) 100,000 neutralizing antitoxin units in horses. A man, surviving 

 a typhoid infection, w^ould still contain in his blood agglutinins for 

 months and years, in spite of the fact that a certain fraction of the 

 antibody would be eliminated (Friedberger, 1902) through urine, or 

 other routes. It was shown (Friedberger and Dorner, 1905) that 

 bacteriolysins are produced in rabbits by 1/1000 of a loopful of 

 cholera vibrios killed at 60°C., and antibodies in rabbits by injecting 

 a total of 0.5-1 .0 mgm. (300,000-900,000) goat red blood cells. 



In the light of some of the above facts concerning the disproportion- 

 ality between the very small amount of antigen used and the many fold 

 quantity of antibody formed, the Biichnerian hypothesis, which con- 

 siders the formation of antibody as a consequence of chemical union 

 between antigen and the antibody, loses its significance (Miiller, 

 1917). Newer and more accurately obtained quantitative data will be 

 presented below concerning this question. 



Seibert (1925) found waters spontaneously became contaminated 

 after standing at least four days under non-sterile conditions. Bacterio- 

 logical tests revealed the presence of chromogenic and non-chromo- 

 genic, motile and non-motile bacteria. Four liters of water on concentra- 

 tion was found to contain 0.036 mg. of nitrogen (after subtracting the 

 blank). She calculated that 1 ml. of fever-producing water would have 

 contained 0.000,000,005 g. of protein. 



Ninhydrin tests on one liter concentrates of highly potent water 

 were negative, as were also the tests on several bacterial filtrates. 



Immunological Studies. A rabbit was injected with 1/50 ml. of 

 water. After an elapse of 12-14 days it was injected with 10 ml. of 

 the same water; within one hour and ten minutes it developed typical 

 shock symptoms, e.g. scratched its nose, fell to one side paralyzed, 

 collapsed, expelled bloody urine, gasped for air and died with violent 

 convulsions. Autopsy revealed an enlarged right heart and congested 

 liver. Several other experiments gave her similar results. Immunization 

 of the rabbits with traces of solid material present in 20 ml. of non- 

 sterile distilled water with a protein content of 0.000,062 to 0.000,12 g. 

 produced an agglutinating serum which was active in dilutions of 

 1 : 8000 against bacteria found in such distilled water. 



Branham and Humphreys (1927) found that the serum of animals 



