388 MEETING OF INSPECTORS 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 your-secretary, but other work came up which interfered with the 
earrying out 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, sent 
me several pounds of very fine wax, such as is used for the manufacture of 
comb foundation. I cultivated the Bacillus alvei upon agar jelly until I 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 humber 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. 
After 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: 
From 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- 
‘ated 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 alvei 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 
