638 



STATE BOARD OF AGRICULTURE. 



All others were kept in the refrigerator. All of these peas except lot 

 A II X mentioned above, were partly shelled one day, then kept until 

 the next day when the shelling was finished and they were canned. 



Within two weeks, spoilage began to occur. The following table gives 

 the data showing the percentage of spoilage in each lot: 



TABLE I 



Per cent spoilage of those autoclaved, .50.9 per cent. 



Per cent spoilage in peas (lot A II X only) autoolaved the same day as shelled, 12.1 per cent. 

 Per cent spoilage of peaa autoclaved 40 min, the next day after shelling (3 lots), 51.9 per cent. 

 Per cent spoilage of peas autoclaved 1 hr. the next day after shclUng (3 lots;, 87. 2 per cent. 

 The high percent of spoilage in the lot autoclaved for 1 hr. in all probability is due to the size of cans used. 

 Per cent spoilaa;e of those cooked in hot water bath, 63.9 per cent. 



Per cent spoilage of those cooked in steam, 73.3 per cent. , • i . 



•Lots A I, II, etc., were processed in the autoclave at 15 lbs. pressure. Lots HWB I, IT, etc., were processed m the hot water 

 bath. Lot S was processed in flowing steam. 



Five cans were selected for examination from lot A I and five from 

 A V in each lot of which the spoilage was complete. 



Each of these cans was examined before opening for evidences of a 

 "swell," i. e., for gas, leakage and any other evidences of spoilage; the 

 can was then carefully washed with a 1-1000 mercuric chloride solution, 

 alcohol was poured over the cover and then burned off, and the can 

 cover lifted only sufficiently to obtain samples of the juice with a pipette. 



Five cubic centimeters of the juice was titrated to determine the 

 acidity. Duplicate sets of dilution plates of 1 to 100, 1 to 10,000 and 

 1 to 100,000 were made. One set of these was placed under anaerobic 

 conditions, using a Novy jar filled with hydrogen. One hundredth of a 

 cubic centimeter was smeared uniformly over one square centimeter on 

 a slide and stained to obtain a direct microscopic count and to determine 

 as well the types of vegetative forms present. Two sets of gelatin agar* 

 shakes were made by the loop. dilution method. One set was heated to 

 80 degrees C. while still liquid for the determination of the presence of 

 spores. 



All cultures were incubated at room temperature. A gelatin agar 

 shake culture was made of each anaerobe isolated. When the anaerobe 

 proved to be in pure culture, transfers were made into two tubes of 

 sterile peas in distilled water.** About one centimeter of sterile white 

 paraflQn oil was poured into one tube of each set in order to give an- 



'Gelatin agar medium: Bouillon cubes, 3 gms.; peptone, lO'gms: salt. 5 gms.; dextrose, 20 pms,; gelatin, 20 gms.; agar, 14 

 gras.; water, 1000 c.c. Adjusted to — 1.5 pei' cent normal torphenolphtalein, or left unadjusted (about neutral) according to 

 needs of organisms. 



•*Pea3 used as a nutrient medium wore prepa-ed by filling test tubes about one-third full with peas taken from a can of ap- 

 parently sterile peas, covering the peas with distilled water and sterilizing in the autoclave. 



