Scientific Proceedings (60). 



might be of interest to those who are called upon to cultivate these 

 organisms. Noguchi's method, although very efficient, offers 

 one great disadvantage, namely that the rubber stopper connection 

 between the upper and lower parts of the apparatus often becomes 

 a source of contamination of the culture. The method I propose 

 consists in utilizing the principles of anaerobic cultivation in 

 general as well as those special features which were worked out 

 by Noguchi. This new method can be used in two different ways. 

 Firstly, one can use the tube as shown on Fig. 2 which instead of 

 having the rubber connection of Noguchi joining its two parts as 

 shown in Fig. I, is made entirely out of one glass tube; but other- 

 wise can be used exactly as Noguchi's apparatus, namely the lower 

 part in which the piece of rabbit kidney is put before the tube is 

 drawn out, is filled with the ascitic broth or sheep serum water up 

 to the point where the tube broadens out again; another piece of 

 tissue is placed in the upper portion of the tube and this tube is 

 filled with the ascitic agar into which the spirochaetse culture is 

 placed. 1 Sterile paraffine oil in a thin layer is placed above the 

 agar and the tube is incubated. Spirochaetes during their growth 

 filter through into the lower portion of the tube exactly as in 

 Noguchi's method. This method is especially convenient when 

 one intends to open the tube many times to examine its contents. 

 The other and better way however of cultivating spirochaetes 

 which does away entirely with the upper part of the tube, 2 

 is the following: I put a piece of tissue at the bottom of the 

 tube; draw it out as before; introduce by means of a capillary 

 pipette the spirochaetse culture and ascitic broth in the lower 

 tube; connect the tube with the vacuum pump, as shown in Fig. 6, 

 warming the lower part of the tube in a water bath at 37 0 to facili- 

 tate the exhaustion of the air; cover the ascitic broth, after ex- 

 haustion, with sterile paraffine oil by means of a special arrange- 

 ment taking advantage of the negative pressure in the tube, and 

 finally seal the lower part of the tube at the point of strangulation 



1 It was found that spirochaetes penetrate into the lower tube more readily if 

 1 per cent agar is used for the ascitic-agar mixture instead of 2 per cent, as recom- 

 mended by Noguchi. 



* A Florence flask with a long neck can be used in place of the tube if a larger 

 quantity of culture is wanted. 



