r>Y K. GREIG-SMlTIi. 83 



A2. The ihst: Al. was separated as the lime salt t'vom the non-volatile acids; 

 the second was prepared directly t'nim the total acids. Al contained 18.12 % 

 ot water of crystallisation, and A2 contained 18.42 %. The latter showed a 

 specific rotation ot [Jp --= — 3.35°, and upon being acidified with hydrochloric 

 acid in the proportion of 2 c.c. of strong acid to 20 c.c. of solution it showed no 

 rotation. The acid was theiefore inactive lactic acid with a laevo-rotatory zinc 

 salt, and this was apparently the only form of acid present. 



In preparing the zinc salt of the B2 acid, three crops of crystals were obtained. 

 The first weighed 2.02 grams, and contained 13. 2G % of water of crystallisation. 

 Zinc jjaralactate contains 12.9 %, equivalent to two molecules. The zinc salt 

 when dissolved in water had a specific rotation of [aJD ^ — 5.18", and with 

 the adilition of 2 c.c. of strong hydrochloric acid to 20 c.c, the rotation became 

 [Jd = -(-2.74°. The firet crop of crystals therefore consisted of paralactate. 

 This acid is said to be contained in meat-extract, but in this ease it was the result 

 of tile bacterial activity, because it was not found in the cultures from Bacterium 

 A which was grown in media prepared fiom the same formula. 



The second croj) of crystals weighed 0.8 grams, and contained 15.18 % of 

 water of crystallisation, showing it to be a mixture of two forms of acid. The 

 third crop weighed 0.30 grams, and contained 18.75 % of water. The zinc salt 

 of ordinary ethylidene or fermentation lactic acid contams 18.18 %, equivi-.lent 

 to three molecules of water, and this was undoubtedly the form of acid in the 

 third crop of crystals. 



The calcium salt was prepared from a portion of the non-volatile acids. It 

 containc<l 26.50 % of water, equivalent to 4J molecules (26.2 %), and was* 

 either a mixture of the calcium salts of the two forms of acid, or it was the 

 more insoluble paralactate, as was indicated by the comparative (fuantity obtained 

 (2.0 grams). The calcium salt of the ordinaiy :i<-i<l would probal)ly have been 

 in the mother liquor from the crystals. 



The acids formed by the two bacteria, A and B, from dextrose in the pre- 

 sence of chalk have been shown to consist chiefly of lactic and acetic acids with 

 small quantities of succinic acid and mixed insoluble fatty acids. Ethyl alcohol 

 was also formed in small amount, and it may be that this was the source of the 

 acetic acid. There was a difference in the nature of tiie lactic acids. Both 

 bacteria formed the ordinary fermentation lactic acid, but B2, in addition, ]iro- 

 duced the dextro-rotatory paralactie acid. 



A Glucoside may he formed. — When the A2 culture was acidifle<l witli sul- 

 phuric acid and extracted with ether, a quantity of fllras was carried uji by the 

 &ther and conveyed to the distillation flask. At the end of the extraction, the 

 fther was shaken up with water and the sui)ernatant ether containing the acids 

 wa.s used for their identification. The yellowish watery liquid was evaporated, 

 and yielded a syrup which was assumed to be glucose carried over with the 

 films. Upon ta-sting it. however, it was found to be intensely bitter. The syrup 

 was diluted with water, acidified with acid and shaken up with chloroform. The 

 chloroform was evaporated off, and a yellow bitter syrup obtained. The acid 

 solution was treated with ammonia in excess and again extracted with chloro- 

 form. Upon evaporating the chloroform, a small quantity of a colourless bitter 

 syrup remained. Tl)e presence of a glucoside is therefore indicated, and should 

 this prove to be correct, the further examination will be dealt with in a future 

 paper. 



