64 BACTERIAL POISONS OF INFECTIOUS DISEASES. 



served twenty-four hours after the poison was added. The quantity 

 of the poison used in the experiments of Madsen varied from 1.2 

 c.c. of a one per cent, solution to 0.2 c.c. of a 0.01 per cent, solution, 

 and a control tube containing no toxin served for contrast. In this 

 way the effects of different quantities of the poison were made plainly 

 manifest to the eye. In those tubes containing the larger quantities 

 of the poison all the corpuscles were dissolved and there remained 

 a blood-red solution, uniform in tint from top to bottom. As the 

 amount of the poison was diminished the number of undissolved 

 corpuscles collected at the bottom of the tube was increased and the 

 color of the supernatant fluid grew less intense towards the top. 

 With the larger quantities of poison the destruction of corpuscles 

 was practically instantaneous and occurred before there was time for 

 subsidence, while in the more diluted solutions the corpuscles parti- 

 ally subsided before they were acted upon by the toxin. It was thus 

 demonstrated that there is a latent period in the action of the poison 

 and that this period increases as the quantity of the toxin is di- 

 minished. It was also found that lowering the temperature increased 

 for a given amount of the poison the latent period. It was also 

 rendered highly probable that the individual corpuscles vary mark- 

 edly in their resistance to the hemolytic action of the poison. 

 Furthermore, it was shown that some corpuscles are more resistant 

 or less resistant to this toxin than to other hemolytic poisons. Thus, 

 corpuscles that were found to be especially susceptible to tetanolysin 

 were less resistant to another hemolytic poison, crotin. 



Madsen continued his interesting work and showed that the 

 hemolytic action of tetanolysin can be neutralized by an antitoxic 

 serum. In these experiments he used a tetanus serum each gram of 

 which contained fifty immunity units against tetanospasmin. This 

 antitoxic serum was diluted to one-half per cent, with a mixture 

 of glycerin and common salt solution. This was designated as his 

 " stock solution " and was further diluted to one-fortieth of one per 

 cent, with physiological salt solution for purposes of experimenta- 

 tion. It was found that for the complete neutralization of two c.c. 

 of the toxin from 1.3 to 1.4 c.c. of the -^^ per cent, antitoxin solu- 

 tion was needed ; i. e., when a mixture containing two c.c. of toxin 

 solution and 1.3 c.c. of the diluted antitoxin solution was added 

 to diluted defibrinated blood, the corpuscles remained unaffected. 

 Following the method pursued by Ehrlich in his studies of the 

 diphtheria toxin, Madsen tried the effect of the partial neutralization 

 of the tetanolysin with antitoxin. As a result of these experiments 

 he found that when 0.10 c.c. of the antitoxin solution (which is one- 

 thirteenth of the amount necessary to completely neutralize two c.c. 

 of the poison) was added to two c.c. of the poison the hemolytic 

 action of the toxin was decreased to one-half its value, and that 

 when 0.25 c.c. of the antitoxin (one-fifth of the amount necessary 



