38 MEETING OF [NSPECTOR8 OF APIARIES. 



would naturally be, supply a healthy colony with comb foundation known to 

 contain the spores and observe the result. This I had hoped to try with the 

 assistance of vour secretary, bul other work came up which interfered with the 

 carrying ou1 of this experiment and consequently it had to be postponed until 

 next year. However, I was able to perform one experiment which throws 

 some light on the subject. Mr. Holtermann, the secretary of your union, senl 

 me several pounds of very fine wax. such as is w<cd tor the manufacture of 

 comb foundation. 1 cultivated the Bacillus alvei upon agar jelly until 1 had 

 a large quantity of the bacilli containing spores; this was carefully scraped 

 off the jelly and dried first in the air and then over sulphuric acid. The result- 

 ing grayish mass was pulverized with a sterilized pestle and mortar, and finally 

 mixed thoroughly with the melted wax kept at a temperature sufficiently low- 

 to prevent the immediate destruction of the spores by heat. By this means an 

 enormous number of spores were introduced into the wax. After stirring the 

 wax for some time in order to insure a proper mixing it was allowed to cool. 

 This, as you all know, takes some time when dealing with a considerable quan- 

 tity. During the cooling I was careful not to disturb the wax. 



Alter it had solidified I set out to discover if I could again obtain my 

 bacillus from the infected wax. If it would germinate in the nutrient media, it 

 certainly would in the bees, and that point was to a certain extent settled. 

 Now I obtained the following results: 



Prom the upper layers of the infected wax I was unable to obtain cultures of 

 the Bacillus alvei, either by melting the wax in the nutrient jellies or by allow- 

 ing particles of the unmelted wax to fall on the surface of these jellies. 



From the under layers, however, the results were different; particles of wax 

 placed on nutrient agar in an oven kept at 98° F. became surrounded in twenty- 

 four hours with a luxuriant growth ^of Bacillus alvei. When the wax was 

 melted into the agar or into beef tea, I also obtained the bacillus, consequently 

 it looks as if the mere fact of enveloping the spores with a film of wax was not 

 sufficient to prevent germination. I confess I can not understand how a spore 

 could germinate when surrounded with a film of wax. Spores in germinating 

 require moisture, and if a spore is completely embedded in wax. it can not 

 obtain sufficient moisture to germinate; I would rather believe, therefore, 

 that in this particular experiment the spores had not each an envelope of wax. 

 but that many of them were partially free from wax. Now. if this was the case 

 in my experiment, where I endeavored to make the incorporation of the spores 

 in the wax as thorough as possible, I certainly think it may frequently be 

 the case when foul-broody wax is used and no particular precaution taken. 

 That even when spores are thoroughly surrounded by wax they may not be 

 freed occasionally by the workers is a point which requires further elucidation 

 and upon which I intend to try some experiments next year. 



In looking through the bee journals, however. I find it everywhere main- 

 tained by foundation makers that they never knew of a case of foul brood 

 originating from foul-broody wax; and I have yet to discover a well authenti- 

 cated case where this has occurred. What explanation can we offer of this 

 widespread opinion? 



I explained to you above that I was unable to cultivate Bacillus <iir< ; from 

 the upper layer of the infected wax. Your secretary also sent me a small 

 specimen of wax which he stated he knew to be from foul-broody comb. This 

 I examined repeatedly for foul brood, but was able to obtain it only once. I 

 think we must look to the physical conditions for an explanation of the free- 

 dom from infection through comb foundation. The difference in. the specific 

 gravity of the bacteria and of melted wax is so great that throughout the 



