TOXINES OF KOCH'S SPIRILLUM. 417 



which quickly diffuse into the medium (and in the experi- 

 ments, through the wall of the sac). By greatly increasing 

 the virulence of the organism, then growing it in bouillon 

 and filtering the cultures on the third and fourth day, they 

 obtained a fluid which was highly toxic to guinea-pigs (the 

 fatal dose usually being -i c.c. per 100 grm. weight). If 

 the dose of the toxine is very large, death follows in an 

 hour or even less. The symptoms closely resemble those 

 obtained by Pfeiffer, the rapid fall of temperature being a 

 striking feature. Post mortem, at the site of inoculation 

 there is a little inflammatory oedema, the internal organs 

 are congested, and the small intestine is distended with 

 fluid contents. The toxicity of the nitrate they found 

 not to be altered by boiling. It is somewhat difficult to 

 reconcile the results of Pfeiffer and Metchnikoff as regards 

 the action of heat, though probably the toxine obtained 

 by Metchnikoff corresponds with the secondary body of 

 Pfeiffer, which he obtained in small quantities. A con- 

 siderable number of observers, however, agree in stating 

 that the toxines obtained from cholera cultures are not 

 destroyed at 100 C. 



A great many observers have attempted to obtain toxines in a 

 chemically pure condition, but so far without results which can be re- 

 garded as conclusive. Hueppe and Wood found that the most active 

 toxines were produced by growing the cholera organism in albumin in 

 anaerobic conditions, and considered that this corresponded to the 

 mode of their production in cholera. Scholl confirmed Hueppe's re- 

 sults, and obtained from cultures under such conditions a peptone which 

 possessed highly toxic properties, and which he called cholera toxo- 

 peptone. These results, however, have been adversely criticised by 

 various observers. Wesbrook obtained different substances according 

 to the media on which the cholera organisms were grown, and yet 

 these produced very much the same effects, chiefly collapse, subnormal 

 temperature, cramps, dyspnoea, etc. Such toxic bodies were even 

 obtained from cultures in asparaginate of soda, which did not contain 

 any proteid substance. He therefore came to the conclusion that so- 

 called toxalbumins etc. are really mixtures of albumins and true toxines, 

 the chemical nature of the latter not having been yet determined. 

 Wesbrook also obtained the toxic bodies in small quantity from the 

 pleural exudate of guinea-pigs killed by the vibrio. Bosc also found 

 that the blood, and to a less extent the urine, of patients who had died 

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