-18- 
At least two anesthetizations of 10 minutas duration spaced a day 
apart are necessary to start egg laying. These may or may not be accom- 
panied by insemination* ^n anesthetic treatment without insemination is 
given by placing the caged queen in a convenient container through irtiich 
carbon dioxide is flowing (fig. s). If the queen is to begin laying at 
the normal age (8 to 11 days after emergence), the second treatment must 
be completed before the seventh day, and preferably before the sixth day, 
because she begins laying 2 to 6 days after the second treatment. Although 
treatments given as early as the second and third day are effective, they 
do not cause laying to start earlier than the normal &ge« 
PROCEDORES FOR TAKTOUS TIPES Of KATINGS AND RESULTS 
TO BE EXPECTED 
Inseminations should be made from the fourth to the tenth day after 
emergence. They have often been made earlier than this (second and third 
day), but at times with poor results. At the other extreme, queens can be 
inseminated at any age if they have not begun laying, but if not made to 
start soon after the tenth day, they will be balled and mistr^ted by the 
worker bees, and the percentage and quality of the laying queens will be 
reduced. 
As much as 10 mm.^ of semen has been given in one operation with suc- 
cess, but because the percentage of sperm that reaches the spermatheca de- 
clines as the load increases, it is better to give several small insemina- 
tions than a single large one. Except for individual matings, a load of 
2 .5 to 4.0 mm. 3 has been found very satisfactory. Results are also less 
variable when several inseminations are made than when an equal amount of 
semen is given in a single insemination. It is best to allow 2 days between 
inseminations for clearing of semen from the reproductive tract, although 
often inseminations on successive days have been quite successful. The 
operation should be performed as rapidly as possible to avoid unnecessary 
exposure of the sperm. 
Individual Matings 
Often it is desirable to mate each queen with a single drone. In this 
case it is important that the maximum possible nimiber of sperms be obtained 
from the single drone used. Therefore, well-matured, well-nourished drones 
should be available, and only those used that seem to ejaculate perfectly 
and to yield a large amovmt of semen heavy with spenn. The best drones 
yield nearly 1 mm.' of semen. Individual matings are usually made only in 
genetic experiments in which it is imperative that the spenn come from but 
a single drone. Therefore, if a series of such matings is to be made, the 
syringe must be sterilized between operations to prevent contamination with 
sperm previously used. In individual matings the percentage of laying 
queens is usually high, and most of them will produce some worker offspring. 
However, so few sperms reach the spermatheca that the supply does not last 
long, and such matings are only made when absolutely necessary. One addi- 
tional carbon dioxide treatment must be given to start egg laying. 
