118 REPORT OF COMMITTEE 



before study, even though freshly isolated from its natural 

 habitat. The procedure to employ is as follows: 



Prepare duplicate sub-cultures in standard glucose broth, and 

 on standard agar slopes, placing cultures of each at 37° and 25°C. 

 On the basis of the resulting growth the organism falls into one 

 of the following series: 



Series I. Organisms which produce good growth (surface 

 growth, distinct turbidity, or heavy precipitate) in twenty-four 

 hours at 37° in glucose broth. 



Series II. Organisms which do not produce good growth in 

 twenty-four hours as above, but do in forty-eight hours at 25° 

 in glucose broth. 



Series III. Organisms which do not grow well in glucose 

 broth but do produce good growth on the surface of agar in 

 twenty-four hours at 37°. 



Series IV. Organisms excluded from the above groups but 

 which produce good growth on the surface of agar in forty-eight 

 hours at 25°. 



Record the series number on the chart at the proper place 

 and proceed with the invigoration by inoculating into another 

 tube of glucose broth for organisms of series I and II, or of 

 standard agar for organisms of series III and IV. Incubate 

 this tube at the temperature, and for the time, called for by 

 the series in which it belongs; then transfer frpm this tube to a 

 third tube and incubate as before. From this third culture 

 make a gelatin or agar plate and incubate at the temperature 

 previously used until colonies of sufficient size for isolation are 

 obtained. Transfer from a typical colony to one or more agar 

 slants and incubate one day at 37° or for two days at 25° accord- 

 ing to the temperature relation of the organism studied. 



In case the organism does not produce vigorous growth on 

 either of these media at either temperature, it should be invig- 

 orated with any medium and at any temperature known to be 

 adapted to its growth. Under such circumstances invigorate by 

 the procedure just outlined but using the medium and tempera- 

 ture found most favorable for the organism in question, record- 

 ing on the chart the method of invigoration adopted. If no 



