THE SERUM TREATMENT OF MENINGOCOCCUS MENINGITIS 791 



serious sequelae, which few survive, is one of the most dreaded of in- 

 fections. 



In administering antimeningitic serum we aim to assist the patient's 

 leukocytes and body-fluids to overcome the infection. Repeated spinal 

 punctures remove portions of the infective material mechanically, but 

 the greatest dependence in bringing about quick destruction of the 

 cocci and effecting recovery of the patient is to be placed upon the serum. 



Preparation of the Antimeningococcus Serum. Method of Flexner and Jobling 

 1. Many strains of meningococcus are used in order that a polyvalent serum may 

 be prepared. Fresh strains from new epidemics and sporadic cases are constantly 

 added. "Fast " strains, or those isolated from cases in which the serum has produced no 

 beneficial effect, are especially desirable. By means of complement-fixation tests 

 Olmstead 1 and her associates have been able to classify meningococci into two main 

 groups; similar results have been secured by Kitchens and Robinson 2 with a protec- 

 tive test. Both tests are apparently highly specific and serve to differentiate menin- 

 gococci from similar microorganisms. 



2. Immunization is performed first with an autolysate of the meningococci, and 

 later with living cultures. 



3. Stock cultures are kept alive by transplanting them every four days in slants 

 of ascitic glucose agar, neutral to phenolphthalein. 



4. In preparing the autolysate, the cultures are subcultured first on glucose-agar 

 slants without serum. After twenty-four hours' growth about 3 c.c. of salt solution 

 are added to each slant, and the culture emulsified. One cubic centimeter is then 

 poured over the surface of glucose-agar slants in large 500 c.c. Blake bottles. After 

 twenty-four hours' incubation heavy, uniform, and diffuse growths are secured. 



Add 10 c.c. of normal salt solution to each bottle and wash off the culture. If 

 necessary, a long heavy platinum loop may be used. Each bottle is tested for con- 

 tamination by staining a smear according to the method of Gram. Each bottle is 

 emptied into a common vessel; 2 per cent, toluol is added, mixed well, and incubated 

 for from eighteen to twenty-four hours. The toluol is then allowed to evaporate, or 

 it may be immediately filtered off through sterile gauze saturated with salt solution. 

 The preparation is kept in a refrigerator and should be prepared fresh every month. 



5. The injections are given subcutaneously about the neck and abdomen. Young 

 and healthy horses are selected for the purpose. The first dose consists of 2 c.c. of 

 the autolysate, and this is gradually increased, depending on the manner in which the 

 animal reacts, until 10 c.c. are given in a single dose. Then inject 2 c.c. of living cul- 

 ture diluted with two parts of salt solution, and increase the doses, the same as with 

 the autolysate, until 10 c.c. of culture are given at one injection. Next inject living 

 cultures and autolysate alternately, until a maximum of from 30 to 35 c.c. are given 

 in one dose; this last is then used as the constant dose until the immunization has 

 been completed. 



Injections are given every five to seven days until the large doses are reached, 

 when they are given every ten days or two weeks. 



Horses usually show quite marked reactions, such as fever, depression, and 

 induration about the site of injection, but if due care is exercised, few animals are 

 lost during the immunization. 



6. Bleedings are usually begun about the fourth month after immunization has 

 been instituted. The horses are bled aseptically from a jugular vein about every 



1 Jour. Immunology, 1916, 1, 307. 2 Jour. Immunology, 1916, 1, 345. 



