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Sir T. Lauder Brunton and Mr. H. Rhodes. 



the kindness of Messrs. Allen and Hanbury, who allowed us the use of 

 their hydraulic press, with a pressure of five tons to the square inch, 

 we were enabled to resume the research. The following was the 

 method adopted : The bone and superfluous fat were removed from 

 the muscular part of a newly killed sheep. The muscle was then 

 minced in a sterilised sausage machine and pounded in a mortar with 

 silver sand. The silver sand was previously cleaned by means of 

 hydrochloric acid and washing with water until all the hydrochloric- 

 acid had been removed. The mass was then put into a canvas bag 

 and placed under the hydraulic press. The juice was received into 

 clean, stoppered bottles, the portion which was yielded on different 

 pressures being received into different bottles. The quantity of juice 

 obtained from a leg of mutton was as follows : — 



1750 grammes of flesh yielded approximately — 



At 0-1 ton pressure per sq. inch ... 450 c.c. of juice. 

 ,, 1*2 tons ,, ,, ... 350 c.c. ,, 



2 "5 tons ,, ,, ... 125 c.c. ,, 



The method of experiment was as follows : — 5 c.c. of the muscle 

 juice were placed in a flask and boiled for one minute, 5 c.c. in another 

 flask remained unboiled. To each flask 50 c.c. of a 1 per cent, diabetic 

 sugar solution and 5 c.c. of a 1 per cent, solution of lactic acid, with a 

 fragment (about 0*25 gramme) of thymol were added. Both vessels 

 were incubated at 37° C. for 24 or 48 hours. After the incubation was 

 finished the sugar was estimated in both flasks by titration with 

 Fehling's solution, after precipitation of the albumin by boiling and 

 neutralisation if required. Six experiments were done with concordant 

 results, and we have only given the result of one as being typical. 



Sugar as estimated by reduction of Fehling fluid — 



1st sample A (boiled juice) 48 hrs.' incubation 0*57 per cent, dextrose 

 2nd „ B (unboiled juice) ,, „ - 2 „ „ 



The destruction of sugar in the flask containing unboiled sugar 

 seemed to be almost certainly due to some glycolytic enzyme, as 

 the contents of the flask remained quite clear at the time of experi- 

 ment. Later on, however, the contents of the unboiled flask became 

 turbid, and after four days a definite growth of fungi was obtained. 

 We next attempted to render the muscle juice sterile by a Pasteur- 

 Chamberland filter. The sugar solution was sterilised by boiling, and 

 all the flasks and other vessels used in these experiments by heating in 

 an autoclave. The muscle juice after filtration was completely sterile, 

 as was shown by the fact that it was kept in a bottle plugged with 

 sterilised wool for many weeks without any bacterial growth exhibiting 

 itself. The glycolytic power of this sterilised muscle juice was tested 

 in the following manner : 5 c.c. of the sterilised juice was placed in 



