40 



ce. serum of rabbit sol. n» 1.— Cont. do- 

 sis: Survived. 



5th Guinea-pig — 1/2000 anliser. — 2 

 cc. serum of rabbit sol. n» 2.— Cont. do- 

 sis: Died tet. 7lh day. 



6th Guinea-pig — 1/1000 antiser. — 2 

 cc. serum of rabbit sol. n" 2.— Cont. do- 

 sis: Survived. 



7th Guinea-pig — 2 cc. ser. rabbit 

 no 1—0,0025 tet. toxin. Survived. 



8th Guinea-pig — 2 cc. ser. rabbit n» 

 2—0,0025 let. toxin. Survived. 



9th Guinea-pig — O.OüOü tet. toxin. 

 Died between 4th and 5th day. 



The rabbit sera together with the 

 dilutions of antitoxin serum were placed 

 for an hour in the incubator at the end 

 of which time the control-dosis of toxin 

 ■was added, it being then allowed to stand 

 for another hour at room temperature. 

 The antitoxin employed v/as the same 

 throughout and titration was always ma- 

 de by the RüSENAU ANDERSON me- 

 thod. 



it thus becomes evident that thirteen 

 days after the last inoculation, there still 

 existed appreciable quantities of horse 

 antitoxin in circulation, although precipi- 

 tins were by then abundant. 



Eight days later, after death of one 

 of the animals, the remaining one was 

 bled, and serum examined. 



Reaction of precipitins present. 



Irst Guinea-pig — 1/2000 antiser. — 2 

 cc. physiol. s. sol. Cont. dosis, Local tet. 

 3rd day, death on 4th. 



2nd Guinea-pig — 1/1000 antiser. — 2 

 cc. physiol. s. sol. Cont. dosis— Healthy, 

 survived. 



3rd Guinea-pig — 1/2000 antiser. — 2 

 cc. serum rabbit n» 1 Control dos. Dia- 

 phragmatic tet. Died afternoon of 2nd 

 day. 



4th Guinea-pig — 1/1000 antiser. — 2 

 cc. serum rabbit n» 1 Control dos. Dia- 

 phragmatic tet. Local tet. evident on 

 Irst day. Found dead next morning. 



.\s will be seen this time, the anti- 

 toxin was entirely eliminated from anti- 

 toxic serum. We were able to ascertain 

 beyond doubt that inactivation for half 

 an hour at 56° does not in any way mo- 

 dify the toxin-inactivaling property, and 

 like DEHNE and HAMBURGER, we were 

 easily able to regenerate antitoxic pro- 

 perty by dissolving the precipitate in as 

 excess of precipitinogen (normal horse 

 serum). Unless the jintitoxin is as.sumed 

 to play the role of a precipitinogen, the 

 interpretation of the phenomenon seems 

 to remain very obscure, seeing that, once 

 it is bound to its corresponding toxin, 

 the precipitating serum is unable to libe- 

 rate the toxin from it's antibody, as we 

 ourselves were able to ascertain. 



Irst guinea-pig — 1/1000 anti.ser. — 2 

 cc. Prccip. ser. — Control dosis. 



2nd guinea-pig — 1/1000 antiser. — 2 

 cc. Control dosis Precip. sera. 



3rd guinea-pig — 1/1000 antiser. — 2 

 cc. physiol. salt. sol. — Control do.sis. 



In the case of the second guinea-pig 

 the toxin, after an hour's contact with 

 the antitoxin, was left for another hour 

 together with the precipitating serum in 

 the incubator. 



The first guinea-pig succumbed to 

 tetanus within two days. The other 

 two survived. Forty two days after 

 the last inoculation, the serum of 

 this same animal, after being bled once 

 more, clearly retained the antitoxin-inac- 

 tivating property. 



Five rabbits were inoculated subcu- 

 taneously with 15 cc. of normal horse 

 serum and bled on 6th, 10th, 16th, 21th 

 and 26th day. 



(Sera used as on pag. 36.) 



Irst guinea-pig — 1/1000 antiser. — 2 

 cc. 6th day-Precip. serum.— Control dosis. 



2nd guinea-pig — 1/1000 antiser. — 2 

 cc. 10th day-Precip. serum.— Control do- 

 sis. 



3rd guinea-pig — 1/1000 antiser. — 2 



