100 HISTORY AND CAUSE OF THE COCONUT BUD-EOT. 



preceding experiments, would distill over, while all the volatile acids would remain 

 behind. The distillate failed to reduce ammoniacal silver solution, thus demonstrat- 

 ing the absence of volatile aldehydes. The distillate did, however, give a powerful 

 iodoform test, showing the presence of alcohol. 



The residue from the distillation was made acid with syrupy phosphoric acid and 

 distilled again to drive over volatile acids. The presence of formic acid was verified 

 in an aliquot part of the distillate by the reduction of mercuric chloride in the presence 

 of sodiimi acetate. The calomel formed was weighed, so that the formic acid con- 

 tained in one cultm-e flask was estimated quantitatively with some degree of accuracy. 

 The 700 c. c of the culture liquid contained 0.197 gram formic acid. 



With ferric chloride the distillate gave a deep blood-red color characteristic of ferric 

 acetate, a verification of the finding of acetic acid. 



The crystalline crusts mentioned in the beginning of this paper seemed to consist 

 mainly of calcium succinate. Oxalic acid could not be found. Lactic acid could 

 not be found. 



The culture liquid still reduced Fehling's solution powerfully. This was at first 

 supposed to be due to the presence of unfermented glucose. However, the presence 

 of formic acid was certainly responsible for a part if not all of this reduction. The 

 culture liquid was not tested for glucose, so that the presence of glucose was not decided. 



Summary: The organism forms much succinic acid and alcohol, as well as appre- 

 ciable quantities of acetic and formic acid. 



REDUCTION OF COLORS. 



Cultures in litmus milk soon turned red and eventually usually- 

 bleached, at least in the lower part. The entire culture never lost 

 its color, but frequently the lower one-half to two-thirds became 

 reduced. 



Cultures in litmus bouillon also reddened and either became 

 almost entirely bleached or partially so. 



Cultures in fermentation tubes containing beef bouillon and 1 per 

 cent cane sugar and litmus became entirely bleached in the closed 

 end, but unchanged in the open end of the tubes, both in the case of 

 Bacillus coli and of the coconut organism. When the tubes are 

 made up with a higher per cent of sugar, for instance, 3 or 5 per 

 cent, the closed end of the tube becomes bleached on steaming and 

 expulsion of the air from that end. According to Dr. Theobald 

 Smith,^ cultures of Bacillus coli grown in these tubes of litmus-sugar 

 bouillon with the bleached closed ends cause the return of the Utmus 

 color. This reaction has not been tried by the writer. 



A series of cultures was made in Dunham's solution (1 per cent 

 peptone plus 0.5 per cent NaCl) and litmus; in Dunham's solution 

 plus indigo carmine, and in Dunham's solution plus methylene blue, 

 both with and \\dthout grape sugar. In the cultures with litmus, 

 in one experiment, the color was reduced only in the tubes contain- 

 ing grape sugar. When cultures were grown in another lot of the 



'Smith, Theobald. The Fermentation Tube. The Wilder Quarter-Century Book, 1893, p. 190. 

 228 



