Mar. i, 1924 
Antisheep Amboceptor and Complement in Fowls 711 
horse, 1 to 50; swine, 1 to 300; rabbit, 1 to 300; guinea pig, 1 to 20; goose, 
1 to 80; duck, 1 to 90; and pigeon, 1 to 90. 
In his hemolytic experiments Rissling used a 5 per cent suspension of 
corpuscles. The serum was obtained fresh. Goose, duck, pigeon, and 
chicken serum caused no hemolysis of sheep cells in amounts of 0.5 cc. 
Chicken serum was found to cause hemolysis of corpuscles of various 
animals when used in the following amounts: Human, 0.3 cc.; swine, 
trace in o. 1 cc., and complete in 0.4 cc. Chicken serum to the amount 
of 0.5 cc. did not hemolyze corpuscles of horse, ox, sheep, guinea pig, 
goose, duck, or pigeon. 
Aschenheim (/) summarized the results of various authors concerning 
the lytic action of the serum of various animals upon the blood corpuscles 
of other animals. In the table incorporated in his article he attempted to 
show the occurrence of natural amboceptor but failed to demonstrate this 
for sheep corpuscles in fresh normal chicken serum. 
Hyde (j), in studying the natural hemolytic antibody of chicken 
serum, found that o. 1 cc. of chicken serum consistently caused complete 
hemolysis of o. 1 cc. of a 1 to 4 suspension of corpuscles of rabbit, guinea 
pig, ox, sheep, and pigeon. The dissolving action, however, was most 
energetic for rabbit cells and least for those of the pigeon. 
Hyde conducted some experiments on the reactivation of heated 
chicken serum. He was unable to do this by the usual procedure, but 
by a modification of the original technic, chicken serum inactivated by 
heat at 56° C. for 30 minutes, was reactivated for rabbit corpuscles with 
nonhemolytic doses of fresh chicken serum to practically its original 
hemolytic titer. The test was made by adding the different amounts of 
heated chicken serum to the tubes, salt solution was then added, followed 
by fresh chicken serum as a complement. The tubes were incubated for 
one hour, the corpuscles were then added and the incubation continued. 
It was found that chicken serum inactivated at temperatures between 
53 0 and 58° C. could be reactivated almost to its original titer by this 
method. Somewhat similar results were obtained by use of guinea-pig 
serum as a complement. 
EXPERIMENTAL DATA 
A flock of 22 fowls was available for this study. All birds were bled 
from the heart. The samples of blood thus obtained were allowed to 
clot and the serum was removed to sterile tubes immediately after cen¬ 
trifugation. 
SEPARATION OF AMBOCEPTOR AND COMPLEMENT 
In order to make titrations of the ambocepter and complement sepa¬ 
rately, it was necessary to absorb the former with washed sheep cells at 
o° C. The various samples of serum were placed in brine at a temperature 
of o° C. A sheep-cell suspension was likewise placed in the bath. After 
sufficient time had elapsed to allow the temperature of the serum and cells 
to be lowered to o° C., 3 cc. of the 5 per cent suspension of cells were added 
to 1 cc. of each sample of serum. The tubes were thoroughly shaken 
and immediately replaced in the bath where they were allowed to remain 
for one hour, which proved to be sufficient time for the absorption of 
hemolysin by the cells. Each sample was centrifuged, and the super¬ 
natant fluid removed to sterile tubes. The sediment was resuspended in 
