METHODS OF STUDYING H.EMOLYSINS. 



into goats without injury. In rabbits of 2 kilos it will hardly be 

 possible to go beyond 100 cc.; and guinea-pigs, corresponding to 

 their weight, proportionately less. According to our experience a 

 single injection of 20-30 cc. sheep, goat, ox, or dog blood leads to a 

 strong formation of haemolysin, which can be still further increased 

 by a subsequent injection of 40-60 cc. six to ten days later. We 

 have found that further injections of the same or larger amounts 

 (80-100 cc.) have no advantage. We have occasionally observed 

 that these were associated with a decrease in the amount of haemolysin. 

 As a rule, the serum attains its maximum power between the sixth 

 and tenth day, 1 but this is subject to individual variations, as is shown 

 by the case described by Ehrlich and Morgenroth of a goat in which 

 an isolysin developed critically on the fifteenth day (see page 29). 



The injections of serum lead principally to the production of 

 antiamboceptors and anticomplements, in some instances also to that 

 of haemolytic amboceptors in consequence of the receptors present 

 in solution in the serum. 2 The production of antiamboceptors 

 necessitates a special selection of the animal species. Our own 

 positive results are limited to the injection of goats either with the 

 serum of a rabbit which had been immunized with ox blood, or with 

 an isolytic serum. Since in these cases the immune serum is toxic 

 for the goat, or, more particularly, acts destructively on the blood, 

 it is necessary to commence with the injection of small amounts 

 (10-20 cc.) and gradually, as the reaction subsides, go on to larger 

 doses. As in the case of all immunizations with toxic substances it is 

 particularly necessary to keep a careful control of the weight of the 

 animals; the rule always to be observed is that immunization can 

 only then be proceeded with when the animal has again attained the 

 weight it originally possessed. 



In order to produce anticomplements larger animals, such as sheep 

 and goats, are injected with increasing amounts of normal serum, 

 beginning usually with fairly large amounts 100 to 500 cc. As a 

 rule, when rabbits have been injected two or three times with guinea- 

 pig, horse, goat or ox serum (commencing with 5 to 10 cc. and increas- 

 ing to 20 to 50 cc.),a plentiful supply of anticomplement will have 

 developed in the serum. In many cases the injection of an inactive 



1 See also Bulloch, Centralblatt f. Bact., Vol. 29, 1901. 



2 See Morgenroth (page 241, this volume) and P. Miiller, Munch, med. 

 Wochensch. 1902, No. 32. 



