104 TOXINES AND ANTITOXINES. 



from decomposed flesh. WLADIMIROFF 1 cultivated the bacilli in 

 Erlenmeyer flasks in an atmosphere of hydrogen, and after seven 

 days added to the culture medium 0*5 per cent, of phenol. He 

 introduced the hydrogen through tubes, which passed through 

 two openings in an india-rubber stopper, and the ends of these 

 tubes were subsequently closed by fusion. DEBRAND 2 asserts 

 that tetanus bacilli grow very well in symbiosis with B. subtilis, 

 and produce powerful toxines. In the case of these bacilli also 

 it has been found that there is not invariably a parallelism 

 between the amount of growth and the toxicity. 



RUPPELL and RANSOM 3 found that the freezing point of the 

 bouillon was somewhat lowered on the production of toxine, 

 which indicates a fresh formation of molecules; and, as the 

 cultivation becomes weaker, the freezing point ought to be 

 slightly raised again. 



USCHINSKY 4 cultivated tetanus bacilli on a proteid-free medium of the 

 following composition : 



Water, lOOO'O 



Glycerin, 



Sodium chloride, 



Calcium , , 



Magnesium sulphate, 



Dipotassium hydrogen phosphate, 



Ammonium lactate, 



Sodium aspartate, . 



30 to 40 



5 7 



O'l 



0-2 to 0-4 

 2 2-5 



6 7 



3-4 



preferably with the addition of 1 to 2 per cent, of grape sugar. The air 

 was excluded by means of liquid paraffin. 



The usual means of sterilisation are employed, especially nitra- 

 tion through a Chamberland filter, and heating to 60 C. 



Tetanus toxine is extraordinarily sensitive to physical and 

 chemical influences. According to BEHRING and KNORR 5 it 

 frequently loses very rapidly a considerable proportion of its 

 toxic power on keeping ; its toxicity is frequently reduced to 

 a hundredth part of the original amount in a few days. 



The atmospheric oxygen, in particular, has a very injurious 



1 Wladimiroff, " Antitoxinerzeug. d. Tet. -Giftes," Zeit.f. Hyg., xv., 405, 

 1893. 



2 Debrand, "Sur un nouveau precede de culture du bacille du te'tanos," 

 Ann. Past., xiv., 757, 1900. 



3 Ruppel and Ransom, "Ueb. Molekularverhaltnisse von Tetanusgift- 

 losungen," Z. f. phys. Ch., xxvii., 109, 1899. 



4 Uschinsky, "Ueber eine eiweissfreie Nahrlosung f. pathog. Bakt.," 

 CentralU.f. Bakt., xiv., 316, 1893. 



5 Behring and Knorr, " Ueb. den Immunisierungswert des Tetanus- 

 heilserums," Zeit.f. Hyg., xiii., 407, 1893. 



