34 
G. P. X o. 4517. Serin obtained by coagulation. 
"Previous treatment : 35 days prior, inoculated subcutaneously with 0.19 c. c. toxine 
Ns. 7—1 unit horse serum Standard B27 .] 
G. ?. 4510. Corpuscles obtained by centrifugation, etc. 
Previous treatment: 35 days prior, inoculated subcutaneously with 0.19 c. e. toxine 
No. 7 — 1 ’uni: horse serum Standard B27 .] 
1 c. c.. -5 per cent corpuscles, 
normal guinea pig Xo. 1 
(washed). 
1 c. c., -3 per cent corpuscles, 
treated guinea pig Xo. 4510 
'washed > . 
' 
1 c. e. of — 
Do. 4-1 
gtt. se- 
rum nor- 
mal 
guinea 
Do. -fl 
gtt. se- 
rum 
suinea 
Pig 
Xo. 4517. 
Do. 4-1 
gtt. se- 
rum nor- 
mal 
guinea 
pig No. 2- 
Do. 4-1 
gtt. se- 
rum 
guinea 
Pig 
Xo. 451". 
Normal ho^se serum Sam. 
0 
0 
0 
0 
0 
0 
Normal donkey serum 
Faintest 
trace. 
0 
0 
0 
0 
0 
Normal aorse serum Sp _ . 
0 
F aintest 
trace. 
Faintest 
trace. 
0 
0 
Faintest 
trace. 
Antitoxic horse serum Xatl. 'HI. 1“ ... 
0 
0 
0 
0 
0 
0 
Antitoxic horse serum Xatl. IX. 19) 
0 
0 
0 
0 
0 
0 
Antitoxic horse serum Xatl. V. It 
0 
0 
0 
0 
0 
0 
Antitoxic horse serum ' Alex. AX-5. Gib- 
son's method . 
trace. 
trace. 
Trace. 
Trace. 
Trace. 
Trace. 
Salt solution (control 
0 
Trace. 
0 
0 
0 
0 
First the corpuscles were placed in the test tubes, then the various 
horse serums, and finally fhe drop of guinea-pig serum. 
The corpuscles were obtained by whipping the blood and washing in 
the usual way four times and suspended so as to make the so-called 
5 per cent suspension. After the corpuscles and the serum were 
mixed they were well shaken and placed in the incubator, again 
shaken at the end of one hour, again at the end of two hours, and at 
the end of three hours taken out of the incubator and placed in the ice 
chest. The readings were taken the next morning. 
This work was repeated several times imtil we were satisfied that 
neither the normal horse serum nor the antitoxic hc-rse serum had 
practically any power of dissolving the hemoglobin from the guinea- 
pig corpuscles. There can therefore be no doubt that there is a toxic 
substance in the blood serum of horses entirely independent of any 
hemolytic action. As a further corroboration we may state that 
autopsies upon guinea pigs which have died from the results of the 
injections of horse serum showed none of the usual effects, indicat- 
ing that hemolysis took place within the body. 
Further, horse serum heated to 60° for six hours is as toxic as 
unheated serum. 
Am a further study of the relation of hemolysis to the toxic action 
and also of the relation of the toxic principle to the haptin group 
we made the following experiments : 
G. ?. No. 249. Four subcutaneous injections of 1.5 c c. each on successive days, using 
normal roan horse corpuscles “ashed three times. The blood, serum of this guinea pig 
developed a high degree of hemolytic power against horse corpuscles. The blood serum 
of this guinea pig was mixed with equal quantities of norma, horse serum and the mix- 
ture injected into the following three guinea pigs: 
