VIII. B, 2 Willets and Schobl: Cerebrospinal Meningitis 137 



Microscopic and macroscopic agglutination reactions were 

 present. On September 13, the twelfth day of the disease, 

 positive microscopic results were obtained in one hour in dilu- 

 tions of 1 to 20 and 1 to 40 of the patient's whole blood. On 

 the following day macroscopic agglutination was present in 

 dilutions of from 1 to 6 up to 1 to 384 of the patient's blood 

 serum, after incubating at 37° C. for two hours and allowing 

 to stand over night. 



Attempts to produce the disease in monkeys and recover the 

 organism in pure culture were unsuccessful. The experiments 

 were as follows: 



On September 9, monkeys 6403 and 6404 were trephined. One cubic 

 centimeter of a thin suspension of the diplococci obtained from a 28-hour 

 culture on Loefler's blood serum was injected beneath the meninges of 

 monkey 6403. Rather profuse bleeding rendered the operation unsatis- 

 factory, since the meninges could not be clearly seen while the injection 

 was being made. Monkey 6404 received 1 cubic centimeter of the 52-hour 

 specimen of cerebrospinal fluid taken on September 7, the flakey material 

 being avoided in so far as possible. The field of operation was clear in 

 this case, the meninges being visible throughout the injection, and hence 

 the operation was quite satisfactory. 



During the following day the monkeys were nervous, uncomfortable, 

 and each had a temperature of 40 °C. On the second day they were greatly 

 improved, and thereafter gradually returned to normal. No convulsions 

 or tremors were noticed in either animal at any time after operation. 



On September 17, 0.2 cubic centimeter and 1 cubic centimeter of a 

 suspension of organisms from several 24-hour Loefler blood-serum cultures 

 were injected into the spinal canal of monkeys 6405 and 6408, respectively. 



Monkey 6408 was found dead at 7 a. m. the following morning. Necropsy 

 was performed at 3 p. m. Smears from 4 different levels of the spinal 

 cord showed Gram-negative, extracellular and intracellular, biscuit-shaped 

 diplococci. Cultures from the heart's blood and the spleen were negative, 

 while those from the four different levels of the cord gave a mixed growth 

 of Gram-negative diplococci and staphylococci. We were unable to secure 

 the diplococci in pure culture. 



Monkey 6405 on the day after the operation was very uncomfortable, 

 nervous, easily frightened, and found repeatedly lying upon its side. 

 However, no tremors nor convulsions were observed. The temperature 

 was 40° C. On September 19, approximately forty-two hours after injec- 

 tion, a small amount (0.2 cubic centimeter) of cerebrospinal fluid was 

 removed by puncture; this gave Gram-negative diplococci in smears, but 

 cultures on Loefler's blood serum were negative. The animal improved 

 daily, and apparently returned to normal. 



On September 27, 1.0 cubic centimeter, 0.6 cubic centimeter, and 0.1 

 cubic centimeter of a suspension of diplococci from several 24-hour Loefler 

 blood-serum cultures were injected into the spinal canal of monkeys 6417, 

 6406, and 6407, respectively. On the following day all three animals were 

 in good condition. The temperature of each was only slightly elevated. 

 Cerebrospinal fluid obtained from each animal approximately eighteen 



