1917 



AMERICAN BEE JOURNAL 



377 



confined in small glass boxes. 



This work done between 1885 and 

 1887 has been discredited. The pos- 

 sibility of accomplishing this feat 

 was again broached by Professor 

 Francis Jager in 1914 and the senior 

 author was asked to co-operate in 

 the work. In the number of "Science" 

 for November 13, 1914, a preliminary 

 report upon a successful case of arti- 

 ficial fertilization was published. 

 Since that time, the writers have at- 

 tempted to repeat the experiment 

 successfully, but have almost uni- 

 formly failed. The advantages, both 

 to the practical beekeeper and to the 

 student of genetics, if this could be 

 done, are obvious and need not be 

 detailed here. 



The queeubee reported upon in 

 "Science" in 1914 wintered in good 

 condition, but soon after removal to 

 the open in the following spring she 

 began to lay drone-eggs as well as 

 worker-eggs and finally produced 

 exclusively drone-eggs. After this 

 had continued for three weeks she 

 was killed and the contents of the 

 spermatheca examined. It was 

 packed with live, active spermatozoa, 

 showing conclusively that fertiliza- 

 tion had taken place. If left alive 

 she would probably have soon re- 

 sumed the production of worker- 

 eggs. 



During the summers of 1915 and 

 1916, 55 duplicates of this experiment 

 were carried out, making a total of 

 63 experiments. Of these, aside from 

 the one already recorded, three were 

 partial successes, the remainder were 

 failures. Twenty-six queens died or 

 were killed by the workers a few 

 days after they were fertilized and 

 before sufficient time elapsed to al- 

 low of egg production. The remain- 

 der did not lay eggs in sufficiently 

 large quantity to ascertain the sex of 

 the progeny or else the queens were 

 killed and opened after a few drone 

 eggs had been laid, and the sperma- 

 theca found to contain no spermato- 

 zoa. 



The method followed in the opera- 

 tion was as follows: Being unable to 



secure successful manipulation by 

 squeezing out the drone organs and 

 allowing the spermatic fluid to drop 

 into the posterior chamber of the fe- 

 male, we carefully dissected out the 

 seminal vesicles of mature drones, 

 using sterile instruments to mix the 

 contents with a drop of sterile salt 

 solution (0.75 gr. to 100 cc H2 0). In 

 the later trials the salt solution was 

 not used, its purpose being merely to 

 dilute the spermatic fluid for greater 

 facility in handling. During cool 

 weather the instruments were kept 

 warm. If the spermatozoa were in- 

 active the material was discarded and 

 a new preparation made. The queen 

 was held loosely between the fingers 

 of the left hand the posterior end 

 upward. With the right hand a cap- 

 illary tube into which the spermatic 

 fluid had been drawn was then in- 

 serted into the Cursa copulatri.x of 

 the queen and the contents gently 

 forced into the vagina by pressure 

 from the mouth. The queen was then 

 placed in a queenless colony and left 

 under normal conditions except that 

 a queen excluder was applied to the 

 hive, between it and the bottom- 

 board. 



To prevent the possibility of nat- 

 ural fertilization, each queen was 

 taken as soon as she emerged from 

 the queen-cell and one or both pairs 

 of wings clipped off. All drones were 

 removed from the hives and a queen- 

 excluder kept on. Queens were 

 taken at all ages, varj'ing from two 

 to thirty-five days, although the ma- 

 jority were treated at the age of 

 six to seven days. During 1916, prac- 

 tically all the queens were thus treat- 

 ed at the time when they were trying 

 to leave the hive, apparently for the 

 mating flight. 



Only mature drones ware used. 

 Careful observations showed us that 

 drones were mature and spermatozoa 

 active when they were five to six 

 day old, but most of those used were 

 ten days or more of age and had had 

 several flights. 



Where queens laid eggs, from 

 eleven to forty days intervened be- 



tween the attempted fertilization and 

 egg laying, with an average of twen- 

 ty days. During 1916, whenever a 

 queen attempted to escape from the 

 hive for a flight she was re-fcrtilizcd. 

 In this way some were operated upon 

 two or three times, but still with no 

 results. 



The three partial successes were as 

 follows: One thirteen-day-old queen 

 was fertilized after the method de- 

 scribed. After an interval of twenty- 

 six days, although the abdomen was 

 enlarged, no eggs had been laid and 

 the female appeared sick. She was 

 killed and the spermatheca exam- 

 ined. It was found quite full of sper- 

 matozoa. The second case was that 

 of a queen whose age was not known 

 at the time of fertilization. Seven 

 days later she was found in the 

 queen trap trying to escape from the 

 hive and was re-fertilized. Several of 

 her eggs produced worker larvae and 

 five eggs were placed in queen-cells. 

 All of these were capped over but 

 robbers destroyed the colony, pre- 

 venting complete observations. 'A 

 third case was a six-day-old queen, 

 which, after an interval of forty 

 days, began to produce eggs, about 

 5 per cent of which produced worker- 

 bees and the remainder drones. Dur- 

 ing the interval between fertilization 

 and egg-laying she three times tried 

 to escape from the hive, and each 

 time was re-fertilized. It was only 

 after the last attempt that her abdo- 

 men began to enlarge and she pre- 

 pared for oviposition. 



If this operation can be done once 

 it would seem that we ought to be 

 able to repeat it with successful re- 

 sults. This led to a closer examina- 

 tion of the female organs. Our ob- 

 servations agree with those of other 

 investigators as to the existence of 

 the S-shaped bend and muscular 

 pump in the spermathecal duct. This 

 probably prevents the forcing of the 

 spermatic fluid into the spermatheca. 

 There is undoubtedly a time or a 

 natural stimulus which causes this 

 valve to open and if the operation 

 can be .performed at this time the 



THIS QUEEN WAS WINGLESS AT BIRTH AND GREAT HOPES VV ERE ENTERTAINED OF HER ACTUAL ARTIFICIAL FERTILIZA- 

 TION 



