49 



Lut it containoil no aiipreciable amount of alcohol. The existence of these cells, 

 however, at this very early stage is of consiilerable interest, and that they were 

 capable of developing rapiilly when placed nnder the projier conditions is shown 

 by the appeaiTince of a sample which was placed near a stove and left till the 

 following day; represented in fig. 2, Plate VII. In this case large numbers of 

 ferment cells are to be seen, of two kinds, one nearly spherical the largest 

 measuring O.OOSl mm. in diameter, and the other longer ami almost cylindrical. 

 The appearance was that of an actively growing yeast. 



The ma-sh wliii.h wa.« left under the usual conditions did not alter thus 

 in appearance. A few more cells may be observed associated with fragments of 

 mycelinm, and with others apparently bursting ami scattering a line dust, but 

 there was no active growth, nor did analy.^is imlieate the formation of any alcohol. 

 Tiie appearance of the mash on the fifth day is shown in fig. 1, Plate VIII. 



After the last sample was taken the moto was heated, and almost immediately 

 a great development of the ferment cells took place. On the seventh day the 

 temfierature was 23°C. and the microscopic appearance (fig. 2, Plate VIII) shows 

 that the cells were budding and growing witli considerable activity, and chemical 

 analysis at the same lime indicated the existence of .5.2 per cent, of alcohol. 

 The diameter of the largest cell wa.s 008.) millimeter and the average size 

 0.0076 mm. The mash on the tenth day bad a very similar appearance to that 

 on the seventh, and on the twelfth, although the temperature was then only 

 lLi°0., the cells still appeared fresh and vigor. ins as in the left of fig. 2, Plate IX. 

 At the same time fragments of the mycelium were to be seen as well as a nmnber 

 of very minute cells, the functions of which are not known. On the fourteenth 

 day the cells had much the t^ame charaetfr as before, the largest still measuring 

 aliout the same. i.e. 0.0082 nun. 



The ne.\t stimple examined was that taken on the seventeenth day, after the 

 further addition of rice and koji, and when the temperature had risen to 19°C. 

 sufficiently high to promote the very active growth of the yeast. Fig. 2 PI. X 

 shows the ajiiTearance of the ferment on that day, and it will be noticed that the 

 size of the cells is rather less, the largest being only 0.0075 mm., perhaps 

 because they were not fidly grown. Py the nineteenth <lay they were again 

 in active growth, and the largest again had a long diameter of 0.0082 mm. 

 The tempenitm-e at that time w;us 2.")-^C. and the amount of alcohol increased from 

 5.8 [Hit cent, on the seventeenth day to Ü.44 percent, on the nineteenth. The growth 

 of the yeast continued, the temperature of the mash on the twentyfir.st day being 

 2G°C., but tliere are to Ix; (observed in the figure of this ma.sh other ferment cells, 

 small stniight or curved filaments which are the cause of the future ileteriuration 

 of the sake. They resemble very closely tlic; filaments which are fomid in 

 "turned" l)eer an<l wine, and are also to 1« found in enormous numbers in sake 

 which has become spoilt. (See figs. 1 and 2 Plate XV^.) 



I iiave drawn also a fil.iment of mycelinm to show that it was slil! [Jivsent, 

 ttltliough as the ferment cell» were very uumeruua and culleeteJ at the surface of 



