The most detailed study of the poison of the honey-bee is that of Josef Langer (1897), who in the course of his work used some 25,000 bees. Various methods of obtaining the active poison for experimental purposes were used. For obtaining the pure secretion, bees were held in the fingers and compressed until the sting was exserted, when a clear drop of the poison was visible at its tip. This was then taken up in a capillary tube or dilute solutions obtained by dipping the tip of the sting into a definite amount of distilled water.

An aqueous solution of the poison was more readily obtained by pulling out the sting and poison sacs by means of forceps, and grinding them up in water. The somewhat clouded fluid was then filtered one or more times. For obtaining still greater quantities, advantage was taken of the fact that while alcohol coagulates the poison, the active principle remains soluble in water. Hence the stings with the annexed glands where collected in 96 per cent alcohol, after filtering off of the alcohol were dried at 40° C., then rubbed to a fine powder and this was repeatedly extracted with water. Through filtering of this aqueous extract there was obtained a yellowish-brown fluid which produced the typical reactions, according to concentration of the poison.

The freshly expelled drop of poison is limpid, of distinct acid reaction, tastes bitter and has a delicate aromatic odor. On evaporation, it leaves a sticky residue, which at 100 degrees becomes fissured, and suggests dried gum arabic. The poison is readily soluble in water and possesses a specific gravity of 1.1313. On drying at room temperature, it leaves a residue of 30 per cent, which has not lost in poisonous action or in solubility. In spite of extended experiments, Langer was unable to determine the nature of the active principle. He showed that it was not, as had been supposed, an albuminous body, but rather an organic base.

The pure poison, or the two per cent aqueous solution, placed on the uninjured skin showed absolutely no irritating effect, though it produced a marked reaction on the mucus membrane of the nose or eye. A single drop of one-tenth per cent aqueous solution of the poison brought about a typical irritation in the conjunctiva of the rabbit's eye. On the other hand, the application of a drop of the poison, or its solution, to the slightest break in the skin, or by means of a needle piercing the skin, produced typical effects. There is produced a local necrosis, in the neighborhood of which there is infiltration of lymphocytes, œdema, and hyperæmia.

The effect of the sting on man ([fig. 27]) is usually transitory but there are some individuals who are made sick for hours, by a single sting. Much depends, too, on the place struck. It is a common experience that an angry bee will attempt to reach the eye of its victim and a sting on the lid may result in severe and prolonged swelling. In the case of a man stung on the cheek, Legiehn observed complete aphonia and a breaking out of red blotches all over the body. A sting on the tongue has been known to cause such collateral œdema as to endanger life through suffocation. Cases of death of man from the attacks of bees are rare but are not unknown. Such results are usually from a number of stings but, rarely, death has been known to follow a single sting, entering a blood vessel of a particularly susceptible individual.

It is clearly established that partial immunity from the effects of the poison may be acquired. By repeated injections of the venom, mice have been rendered capable of bearing doses that certainly would have killed them at first. It is a well-known fact that most bee-keepers become gradually hardened to the stings, so that the irritation and the swelling become less and less. Some individuals have found this immunity a temporary one, to be reacquired each season. A striking case of acquired immunity is related by the Roots in their "A B C and X Y Z of Bee Culture." The evidence in the case is so clear that it should be made more widely available and hence we quote it here.

A young man who was determined to become a bee-keeper, was so susceptible to the poison that he was most seriously affected by a single sting, his body breaking out with red blotches, breathing growing difficult, and his heart action being painfully accelerated. "We finally suggested taking a live bee and pressing it on the back of his hand until it merely pierced his skin with the sting, then immediately brushing off both bee and sting. This was done and since no serious effect followed, it was repeated inside of four or five days. This was continued for some three or four weeks, when the patient began to have a sort of itching sensation all over his body. The hypodermic injections of bee-sting poison were then discontinued. At the end of a month they were repeated at intervals of four or five days. Again, after two or three weeks the itching sensation came on, but it was less pronounced. The patient was given a rest of about a month, when the doses were repeated as before." By this course of treatment the young man became so thoroughly immunized that neither unpleasant results nor swelling followed the attacks of the insects and he is able to handle bees with the same freedom that any experienced bee-keeper does.

In an interesting article in the Entomological News for November, 1914, J. H. Lovell calls attention to the fact that "There has been a widespread belief among apiarists that a beekeeper will receive more stings when dressed in black than when wearing white clothing. A large amount of evidence has been published in the various bee journals showing beyond question that honey-bees under certain conditions discriminate against black. A few instances may be cited in illustration. Of a flock of twelve chickens running in a bee-yard seven black ones were stung to death, while five light colored ones escaped uninjured. A white dog ran among the bee-hives without attracting much attention, while at the same time a black dog was furiously assailed by the bees. Mr. J. D. Byer, a prominent Canadian beekeeper, relates that a black and white cow, tethered about forty feet from an apiary, was one afternoon attacked and badly stung by bees. On examination it was found that the black spots had five or six stings to one on the white. All noticed this fact, although no one was able to offer any explanation. A white horse is in much less danger of being stung, when driven near an apiary, than a black one. It has, indeed, been observed repeatedly that domestic animals of all kinds, if wholly or partially black, are much more liable to be attacked by bees, if they wander among the hives, than those which are entirely white."