LABORATORY AND GREENHOUSE STUDIES. 93 



nearly the same amount, respectively, except coconut No. 3, which 

 showed as much as Bacillus coli. 



Other cultures of the organism were made in ordinary bouillon, 

 and an attempt was made to separate indol and phenol, if present, by 

 distillation. No results were obtained, either by the sulphuric-acid 

 and sodium-nitrite test for indol or by the Millon's reagent and the 

 ferric-chlorid test for phenol. These experiments were repeated 

 several times, and the same results were obtained. It would seem, 

 therefore, that a small amount of indol may be produced, but no 

 phenol. 



PRODUCTION OP HYDROGEN SULPHID. 



Cultures of the coconut organism made in an iron-peptone solution 

 had in a week's time a slightly or wholly blackened precipitate, 

 and the solution was either inclined to be a greenish black or was 

 intensely green and black, thus indicating the production of hydro- 

 gen sulphid. Lead acetate paper used for testing the solution became 

 discolored, also indicating the presence of hydrogen sulphid. 



Cultures were also made directly in a lead acetate solution with 

 peptone and showed a good growth. The precipitate in all of the cul- 

 ture tubes was black, indicating the production of H2S, while in the 

 check tube the precipitate was wMte. These cultures were also 

 tested with lead acetate paper, which showed the brown-black discol- 

 oration typical of HjS. 



PRODUCTION OF AMMONIA. 



A 250-c. c. flask containing 100 c. c. of beef bouillon + 15, was inoc- 

 ulated with the organism and incubated for 18 days. The culture 

 was then distilled with the addition of 2 grams of calcined magnesia, 

 and to 50 c. c. of the distillate was added 1 c. c. Nessler's solution. A 

 bright orange-yellow color was produced. Checks were made by dis- 

 tilhng over uninoculated bouillon which gave a gray-black color with 

 Nessler's solution and by the use of solutions of ammonium hydrate, 

 1 to .1,000, 1 to 5,000, 1 to 4,000, and 1 to 3,333i All of the solutions 

 containing ammonia gave an orange color on the addition of Nessler's 

 solution. The color of the reaction of the culture most nearly corre- 

 sponded to the check solution containing 1 to 4,000 of ammonia. 



Cultures were made in Fischer's solution, plus 1 per cent dextrose, 

 plus 1 per cent KNO3. The solution contained dipotassium phosphate, 

 magnesium sulphite, and calcium chlorid. The growth of the organ- 

 ism after three weeks was fair. The culture was distilled over and 

 tested for ammonia. The distillate showed the presence of a very 

 small quantity of ammonia, about 1 to 80,000. Unfortunately, how- 

 ever, a check flask on being distilled over also showed about the same 



228 



