-2A- 
during confinement seem to be an aid rather than a hindrance. Its pre- 
sence seems to increase the pressure inside the abdomen when the muscles 
contract, assuring good eversion of the penis and ejaculation, it has 
been noticed that drones whose flight has not been restricted by confine- 
ment to the hive will ejaculate better and in greater numbers after they 
have been kept in a cage for a day or two vdth workers, sugar sirup, and 
pollen. Drones do not live so long when coni'ined as when they are able 
to fly naturally. In the nursery colonies some of .them die soon after 
they reach the age of 25 days, and few reach 35 days. 
Three important requirements of the drone nursery colony are ample 
bees, ample pollen, and queenlessness. Drones need the care of worker bees, 
^hen kept in cages and provided only with pollen, water, and sugar syrup, 
they soon die, but when about twice their number of worker bees are added, 
they live to sexual maturity and contain a good quantity of usable semen, 
rthen pollen is omitted drones live for some time, but very few sperms 
develop. ><ueenlessnes8 is also necessary when a large number of drones 
are present; they live longer and mature better in queenless colonies. 
In 1944 the young drones in two queen- right nursery colonies were noticed , 
massed on the bottom board with not a single one on the combs. After the 
queens had been removed long enough for the bees to realize their queenless- 
ness, all the drones were found on the combs apparently being well cared 
for. 
SIMdARY 
By modification of the equipment and methods of earlier workers on 
artificial insemination of queen bees, the technique has been improved suf- 
ficiently to make its use practical in bee breeding as well as in genetic 
studies. In this manual the important features of the anatomy of the sex 
organs, the construction of special equipment, and the insemination proce- 
dure are described. The essentials of the method are as follows i 
(l) The semen is taken from the everted penis after the drone has 
been placed in chloroform fumes to induce partial eversion and the abdomen 
squeezed to continue the eversion imtil the semen has appeared at the end 
of the penis. 
{Z} The semen is deposited in the oviducts in natural mating. To 
do this it is necessary to push aside a tonguelike projection, from the 
ventral wall of the reproductive tract, called the valvefold, so that the 
point of the syringe can roach the mediaji oviduct opening. 
(3) The syringe tapers at the end so that its point is small enough 
to enter the median oviduct, while the main barrel is large enough to have 
a practical capacity. 
(4) At least two inseminations are given at 2-day intervals to approx- 
imate the normal number of sperm in the spermatheca. 
(s) Carbon dioxide is used as an anesthetic. This gas not only 
relaxes the queens, but also stimulates them to begin laying. Two anesthe- 
