402 Cerebro-spinal Meningitis 



while the spinal puncture is made, as shown in Kolmer's illustration. 

 The fluid is allowed to drop into sterile test-tubes or vials with sterile 

 stoppers. From 5 to 15 cc. should be withdrawn. No ill effects 

 have been observed from the operation." 



In making a culture from this fluid Park points out that, as 

 many of its contained cocci are dead, a considerable quantity of the 

 fluid (say about i cc.) must be used. 



The cocci can also be cultivated from the nasal discharges as 

 was shown in 6 cases of cerebro-spinal meningitis studied by Weich- 

 selbaum, and in 18 studied by Scherer. Elser* isolated the organism 

 from the circulating blood of patients suffering from that form of 

 epidemic cerebro-spinal fever known as "spotted fever" in which 

 there is a purpuric exanthema. 



Cultivation. — The meningococcus though successfully cultivated 

 by Weichselbaum is not easy to cultivate and disdains most of the 

 usually employed media. It is aerobic. Growth takes place within. 

 a narrow temperature range 25°-4o°C., the optimum temperature 

 being 37°C. In handling cultures of any kind, great care should 

 be taken to prevent them from becoming chilled. Cultures that 

 have been growing well, sometimes fail to continue when taken from 

 an incubation oven to a cold room for a short time for transplanta- 

 tion. It develops very slight growths upon agar-agar and glycerin 

 agar-agar. Growth is better upon agar-agar containing ascitic fluid 

 containing i per cent, of dextrose and upon Lofiler's blood-serum 

 mixture. According to Goldschmidt,t it can grow upon potato, 

 though most investigators faU. to find any development upon this me- 

 dium. It does not grow in gelatin. It does not grow in plain bouil- 

 lon but when a little calcium carbonate is added to neutrahze any 

 acid formed, and i per cent, of dextrose and some ascitic fluid or 

 sheep-serum added, the broth becomes an excellent medium. The 

 cultures are usually scanty and without characteristic features. 



FlexnerJ found that the difficulties of cultivation were greatly 

 reduced by the employment of sheep-serum water prepared according 

 to the method of Hiss (sheep-serum i part, water 2 parts, sterilized 

 in the autoclave) and mixed with a beef-infusion agar-agar contain- 

 ing 2 per cent, of glucose. The quantity of sheep-serum need not 

 exceed Ko to >f of the volume of the agar-agar. It is added to the 

 sterile melted agar, which is afterward slant€d in test-tubes and 

 allowed to congeal. But by far the best medium for the isolation and 

 cultivation of the meningococcus is slightly alkaUne (+0.5) agar- 

 agar, containing i per cent, of dextrose and i per cent, of laked 

 human blood. (See directions for making the medium in the chap- 

 ter upon Culture Media.) Upon this medium the cocci rarely 

 fail to grow, whether taken from the nasal secretions, the cerebro- 



* "Jour. Medical Research," 1906, xiv, 89. 



t "Centralbl. f. Bakt. u. Parasitenk.," 11, 22, 23. 



t "Jour. Experimental Med.," 1907, DC, p. 105. 



