Isle of Wight Disease in Hive Bees. 41 
IIlL—INVESTIGATIONS BEARING ON THE RELATION OF 
NOSEMA APIS TO ISLE OF WIGHT DISEASE. 
METHODS oF EXAMINATION OF BEES FOR THE PRESENCE OF NOSEMA APIS. 
As carried out by Joun Innzs, B.Sc., M.B., Captain R.A.M.C. 
Each bee was examined in a routine manner, and the following notes 
were made :— 
(1) Activity, as determined by movements in the postal cage—usually 
a match-box. 
(2) Power of flight on exposure in open space. 
(3) Dislocation of wings; if present or absent. 
(4) Gait; whether co-ordinated or paretic. 
(5) Response to stimuli, such as handling, etc. 
The chyle-stomach and colon of each bee were examined systematically, 
and the removal of these parts from the body was carried out in the following 
manner. The head of the bee was firmly crushed between fairly stout 
dissecting forceps, and while held thus, the last segment of the abdomen 
was grasped with straight fine-pointed forceps, and gentle traction applied. 
In this way the colon was carefully pulled out and the chyle-stomach also 
came away with it. These organs were placed on a slide, separated at 
their anatomical junction, and teased. Cover-glasses were then applied, and 
the preparation examined microscopically in the wet, unstained condition. 
If the spores of Nosema apis were present they were readily identified by 
their shape, size, and refractility. Young, intracellular stages and planonts 
were also observed in like manner. 
Stained preparations from smears of the colon, and of the chyle- 
stomach, were made from every sample of bees examined, and the stain 
which was almost exclusively used was the iron-hematoxylin of Heideinhain. 
Although requiring longer time for staining, this was found to give the 
most reliable and most definite results. The fixative used was a hot alcoholic 
solution of corrosive sublimate. 
Various other stains were tried, such as Giemsa, Romanowsky, Erhlich’s 
triacid, etc., but the varying results led to their abandonment in favour of 
the more stable hematoxylin preparation. 
No stain, however, was found which would show up the spores clearly. 
Various experiments in this connection were tried, but with no success. 
Heat had very little effect in helping the stain to penetrate the resistant 
spore capsule. For its identification in stained preparations we could 
