662 



SCIENCE 



[N. S. Vol. XLL No. 1061 



The object of this investigation was to devise a 

 simple, practical and reliable method for the iso- 

 lation and cultivation of the tubercle bacillus from 

 the sputum and feces. Most of the methods em- 

 ployed in the last twenty years do not give uni- 

 formly positive results. 



Taking into consideration the inhibitory action 

 of gentian violet on many organisms, it was se- 

 lected as the most favorable stain. 



Preparation of the Media 



I. Meat Juice. — Five hundred grams of beef 

 or veal are infused in 500 c.c. of a 15-per-eent. 

 solution of glycerine in water. Twenty-four hours 

 later the meat is squeezed in a sterile meat press 

 and collected in a sterile beaker. 



II. Eggs. — Sterilize the shells of the eggs by 

 immersing for ten minutes in 70 per cent, alcohol. 

 Break the eggs into a sterile beaker, mix well and 

 filter through sterile gauze. Add one part by vol- 

 ume of meat juice. 



III. Gentian Violet. — Add sufficient 1 per cent, 

 alcoholic gentian violet to make a dilution of 1 to 

 10,000. 



Place in sterile test tubes and inspissate for 

 three successive days. First day at 85° C. until 

 all the medium is solidified. On the second and 

 third days not more than one hour at 75° C. 

 Method of Isolating Tubercle Bacilli from Sputum 



The use of fresh sputum is advisable. A mix- 

 ture of the sputum and a 3-per-cent. sodium hy- 

 drate solution are left in the incubator for 20-30 

 minutes, then neutralized to sterile litmus paper 

 with normal hydrochloric acid, centrifugalized and 

 the sediment inoculated into the tubed media. 



Method of Isolating Tubercle Bacilli from the 

 Feces 



The isolation of tubercle bacilli from the feces 

 is not an easy problem. The concentration of the 

 sodium hydrate is not as important as the length 

 of exposure. The solid food particles are removed 

 from the feces by dilution with water and filtra- 

 tion through gauze. 



The filtrate is saturated with sodium chloride 

 and at the end of half an hour all the bacteria will 

 be found floating in a fine film. This film is col- 

 lected and normal sodium hydrate added, shaken 

 well and incubated at 37° C. for 3 hours. Then 

 neutralized as is sputum, centrifugalized and sedi- 

 ment inoculated. 



The method presented has proven very simple 

 and accurate for the isolation of tubercle bacilli 

 from the sputum. The partial success in isolating 

 and cultivating tubercle bacilli from the feces may 



be due to the fact that many of the bacilli are 

 possibly dead. 



Comparative Analysis of Several Peptones: E. C. 



COLWELL. 



An investigation of the comparative merits of 

 four brands of peptone is being made to deter- 

 mine the advisability of substituting an American 

 brand for Witte's in Standard Methods of Bac- 

 teriological Analysis. The following table em- 

 bodies the results of the chemical analysis. 



The tests made upon the peptones to determine 

 their relative reliability in the making of culture 

 media has not yet covered long enough time to 

 warrant any definite conclusions. However, it may 

 be said that those peptones containing lactose 

 seem to be inferior to those which are free from 

 lactose. 



A New Method of Precipitating Cellulose for Cel- 

 lulose Agar: F. M. Scales. 

 The method of precipitating cellulose to be used 

 for the preparation of cellulose agar is as follows : 

 100 c.c. of concentrated sulphuric acid are diluted 

 with 60 c.c. of distilled water in a two-liter Er- 

 lenmeyer flask. The diluted acid should be cooled 

 to about 60° to 65° C. Moisten with water five 

 grams of filter paper, which are sufficient for one 

 liter of cellulose agar, and add it to the acid, 

 which should be vigorously agitated until the cel- 

 lulose is dissolved. The flask is then filled as 

 quieky as possible with cold tap water. The proc- 

 ess of dissolving the paper and filling the flask re- 

 quires about one minute. The precipitate may now 

 be thrown on a filter and washed with distilled 



4 Lactose, about 10 per cent. 



