10 THE BACTEEIA OF THE APIAEY. 



corded dimensions should always be accompanied by facts concerning 

 the medium, age, and temperature of incubation. The measure- 

 ments recorded in this paper were all taken of organisms in prepara- 

 tions made from a 24-hour agar culture stained with carbol-fuchsin. 

 The involution forms are not reckoned in the results. 



(5) Spores. — The presence of spores was determined in each case 

 by staining the various cultures at different ages. A check was made 

 on their presence by means of the thermal death point. 



(c) Flagella. — Loeffler's method, as modified by Johnson and 

 Mack, was used for staining the flagella (2). 



{d) Motility. — Motility may be present in cultures when first iso- 

 lated, but after artificial cultivation appear to be entirely lost. The 

 reverse of this also may be noted. No cultures should be recorded 

 as nonmotile until cultures on various media at different temperatures 

 and of different ages shall have been studied. Hanging-drop prepar- 

 tions were made from cultures on agar and bouillon, both incubated 

 and not incubated, and on gelatin. 



(e) Staining froperties. — Basic carbol-fuchsin was the stain used 

 almost exclusively. In the use of Gram's staining method, carbolic 

 gentian violet (5 per cent carbolic acid 20 parts, saturated alcoholic 

 solution gential violet 2 parts) was applied to a cover-glass prepara- 

 tion from a 24-hour culture on agar for 5 minutes, placed in Lugol's 

 solution 2 minutes, and placed, without rinsing, in 95 per cent alcohol 

 for 15 minutes, removed, washt in water, and allowed to dry. 



(/) Oxygen requirements. — Determinations were made by ob- 

 serving whether a growth took place in the closed or open arm or 

 both, of the fermentation tube containing glucose bouillon. 



Media Employed and Suggestions as to tlie Description of Cultures. 



{a) Bouillon. — All bouillon used was made from beef (meat 1 

 part, water 2 parts) , to which infusion 1 per cent Witte's peptonum 

 siccum and one-half per cent sodium chlorid were added. The re- 

 action of the solution was then determined by titrating, and made 

 -j-1.5 to phenolphthalein. 



In describing a culture growing in bouillon as a medium, there 

 is usually a more extended description given than in the case of 

 sugar and sugar-free bouillons, since cultures in these media do not 

 differ materially in gross appearance from those observed in the 

 plain bouillon. 



(6) Sugar-free houillon. — This bouillon is made free from sugar 

 by the use of B. coli communis, after which peptone and sodium 

 chlorid (NaCl) were added as in bouillon. 



(c) Sugar bouillons. — Five different sugars — glucose, lactose, sac- 

 charose, levulose, and maltose, as well as mannite — were used in the 

 study. If a 1-per-cent solution of glucose in plain bouillon Avas fer- 



