and Laboratory Methods. 



1807 



Fig. 50. 



large, otherwise the anaerobic properties of the 

 fluid in the closed branch may be less effective. 

 Lastly, the angle formed by the two branches of 

 the tube must not be too acute, otherwise the 

 tube must be tilted so much during the trans- 

 ference of the fluid from the tube to the closed 

 branch that there is danger of its moistening the 

 plug or even running out of the bulb." 



Method. — Heat the fermentation tube con- 

 taining peptonized, sterile glucose bouillon in the 

 steam sterilizer, cool, and inoculate it with the 

 culture in question. In case of a pure anaerobic 

 culture the growth will take place in the closed 

 bulb and the line of demarcation between the 

 turbid, teeming liquid of the closed branch and 

 that of the bulb and connecting tube is sharply 

 drawn. 



In 1899 Smith recommended a slight modifi- 

 cation of the above method, using the same fer- 

 mentation tube. 



Method. — Kill a guinea-pig, rabbit, pigeon or 

 other small animal with chloroform ; tear pieces 

 of the internal organs, more particularly of the 



spleen, liver, and kidneys, as large as peas or beans from the organs with sterile 

 forceps and quickly introduce them into the fermentation tubes, containing 

 ordinary sterile, peptonized bouillon. The tissue should be eventually forced 

 into the closed branch of the tube with a sterile platinum wire. A series of 

 tubes are prepared at one time and placed in the incubator for several days to 



reveal any contaminating bacteria from the 

 air or from the introduced tissue. Tubes 

 provided in this way with bits of sterile 

 tissues furnish most favorable conditions 

 for the cultivation of anaerobes. They may 

 be kept indefinitely, and when partly dried 

 out they may be refilled with sterile water. 

 Anaerobes will still multiply freely in them 

 though they have not been reboiled. In 

 fact, boiling would cloud the bouillon by 

 coagulating the albumin from the intro- 

 duced material. It is frequently very de- 

 sirable to have on hand fluid cultures of 

 anaerobic species for the study of morpho- 

 logical and physiological characters, a fact 

 which makes this method especially valu- 

 able. 

 Fig. 51. Smith constructed two more apparatus 



F 



