xiii, b, 5 Panganiban and Schobl: Cholera Stool Specimens 277 



EXPERIMENTS II AND III. SALT SOLUTION AND OX BILE 



Five cubic centimeters of sterile sodium chloride solution, in 

 concentration of 0.5 per cent, 1 per cent, 2 per cent, 5 per cent, 

 10 per cent, 15 per cent, 20 per cent, 25 per cent, and 30 per 

 cent, were placed into a set of sterile test tubes, and 5 cubic 

 centimeters of 50 per cent, 75 per cent, and pure ox bile were 

 put into the tubes of the second set. In both of these sets one 

 tube containing 5 cubic centimeters of sterile normal salt solu- 

 tion was used as a control. 



About 10 grams of normal stool were ground up in 100 cubic 

 centimeters of sterile normal salt solution, and the mixture was 

 filtered through cheesecloth. Ten cubic centimeters of sterile 

 normal salt solution were added to each of four 24-hour-old 

 agar cultures of cholera vibrio, which were then washed, and 

 the suspension was added to the filtered emulsion of the stool. 

 The final mixture of cholera vibrios and stool was thoroughly 

 shaken in flasks containing glass beads. Five cubic centimeters 

 of this suspension were then added to each one of the tubes 

 containing varying concentrations of salt solution and ox bile, 

 respectively. The tubes, after the addition of cholera stool 

 suspension, were thoroughly shaken and were kept at room tem- 

 perature (32° C). Immediately after mixing, one peptone 

 water culture was planted from each of the tubes by transferring 

 three loopfuls of the stool emulsion. After twenty-four hours' 

 incubation Dieudonne's plates were planted from the peptone 

 culture. At the end of another twenty-four hours' incubation the 

 colonies that developed were examined microscopically. Smears 

 were made, and miscroscopic agglutination was performed. 



Similarly peptone water cultures, Dieudonne's plate, and mi- 

 croscopic examinations of the colonies were made every day 

 during the first week, after which time examinations were made 

 once a week only. It is evident from Table II that sodium 

 chloride preserved the cholera fasces successfully for the period 

 of five weeks, at least, in concentration of from 0.5 to 5 per 

 cent. In concentration higher than 5 per cent the vibrios could 

 not be found after five and four days, respectively. 



In Table III the results of the experiment, in which bile was 

 used as preservative, are tabulated. It shows successful pre- 

 servation of cholera faeces in dilution of 50 per cent, 75 per cent, 

 and 100 per cent of bile with normal salt solution as a control. 



In the previous experiments the sodium chloride solution and 

 ox bile gave equally good results as preservatives of cholera 

 stools for delayed examination when the amount of cholera 



