228 Journal of the Department of Agriculture. 



infected, before these bacteria have had time to produce toxin, and 

 the disease only occurs when the organisms grow saprophyticaily 

 upon dead organic matter. 



The organisms themselves belong to the anaerobic group, i.e. 

 they only grow in absence of air or in association with aerobes which 

 take up such oxygen of the air as finds access to the putrefying 

 material, or in association with oxidizing enzymes and easily oxidiz- 

 able organic matter (e.g. in liver tissue exposed to air). The fact 

 that they grow in mixed culture with other organisms explains why 

 it is that carcass m^tterial exposed to the atmosT)here becomes toxic. 

 It must not be imagined that they are the cause of putrefaction. 

 They may possibly contribute to decomposition, but the real stinking 

 putrefaction is the woik of the other common bacteria with which 

 they are associated. 



As just s+ated. they are sporulating ore-anisms, and although 

 toxic material may be rendered non-toxic by boiling for a few minutes 

 it may become toxic airain after the undestroyed "-pores have germi- 

 riated out. To sterilize toxic material completely it is necessary 

 either to heat under steam pressure at about 180° C. or to boil on 

 each of" three successive days. 



So far the causal organism has not been isolated in pure culture, 

 although that may be accomplished at any moment now. The 

 technical difficulties in the way are ronsiderable. Impure toxic 

 fultures, however, are easily obtained, in which three different sporu- 

 lating organisms are ronstantly present. Two of thpse dominate the 

 fultures, but when taken alone produce no toxin. The predominant 

 one appears to be B. pvfrificiis. a common soil anaerobe. The 

 second dominant organism is a ''racquet" form. The third is of 

 " drum-stick " form, and is the one to which suspicion attaches at 

 the moment. 



Since the different members have not yet been completely separated 

 it is not yet possible to state which is responsible for toxin production. 

 Indeed, we are rather inclined to think that two. or all three, are 

 essential: that ordinary putrefactive bacteria prepare the medium 

 for the particular bacteria which actually produce the toxin. At first 

 we were inclined to consider that we were dealing with B. hofulinrnt, 

 but comparison of the physiological effect of lamziekte toxin with the 

 toxin from two different strains of botulinus excludes identity. 



All these details will be cleared up in tim.e, but meanwhile the 

 more urgent economic experiment's reouired for the elucidation of 

 practical points, are being pressed with the highly virulent toxin 

 producpd in mixed cultures. As already mention'^d. this toxin is so 

 powerful that a quantity of 0.0001 c.c, or a few millionth parts of 

 an ounce per kilogram body-weight, injected under the skin, is 

 sufficient to produce lamziekte in cattle. The dosage required to 

 produce the disease experimentally in other animals such as horses, 

 sheep, goats, rabbits, guinea-pigs, ostriches, and poultry, varies 

 according to the species of the aiiimal. The minimum lethal dose 

 produces typical lamziekte in cattle in about five days, but the period 

 can easily be reduced to within twenty-four hours by sufficiently 

 increasing the dose. 



Once it had been established that lamziekte i< ju-oduced by a 

 toxin elaborated bv bacterial action in carcasses undergoing putre- 

 faction: that in all probability it is produced in no other way: and 



