Hivemaker.
Queen Bee
Had a link passed on to me about this subject by Norton,have since looked into it a bit more, and believe it could be very useful in queen breeding,depending on success rate,has anyone else tried this.
1. Preparation of drones/semen:
Let the queen lay unfertilised eggs in drone combs about 40 days (24 days development + ca. 16 days for sexual maturity)before AF. These drones should hatch under controlled conditions to be sure about their origin. If e.g. marked with colour paint on their thorax, they can be allowed to freely fly because only a few are needed (one?).
Here comes the most difficult part:
Fill a sterile syringe (volume about 1 ml) about half with sterile sperm dilution buffer (same as used as a stop solution in AM). Attach a sterile glass capillary (about same as with AM) to the syringe and make the dilution buffer fill the capillary, then draw back to make a small volume of air enter the capillary. This air bubble is used to separate the semen from the dilution buffer (this large portion in the ‘back’ of the syringe). The semen of one selected drone (about 1 ul) is then collected in the glass capillary (same procedure as with AM). Thereafter, draw about eight to ten times the volume of the semen (i.e. 10 ul) of semen dilution buffer into the syringe and mix the semen and this small volume of buffer through repeated draw and push cycles on a sterile glass plate. The prepared, diluted semen can be used several hours if stored at room temperature and in the dark.
2. Preparation of unfertilised eggs:
In the morning, cage the queen on one side of a empty fully drawn drone comb. It takes some time till she begins egg laying. In the afternoon transfer the queen to the other side of the same comb. Now, she will continue egg laying after a few minutes. These eggs can now be used for AF. (My comment: In another article I read that if unfertilised eggs will enter development spontaneously after about 4 hours. So in practice, one could wait about 2-3 hours before taking the drone comb to ensure that enough eggs were laid).
The syringe with the diluted sperm is pushed so that the diluted sperm forms half a droplet at the end of the glass capillary. A egg to be fertilised must now be covered with diluted sperm at it’s upper 25 % (the free end of the egg not being attached to the cell)for a second. That’s it! To prevent the sperm from drying, the droplet is drawn back into the syringe each time after AF. Be sure to mark the respective cells (using e.g. an overhead transparency) on the comb.
The comb with AF-eggs is then transferred to a previously dequeened colony. After 3 days the larvae can be grafted as usual. The raised queens can be used for AM too, of course.
If queens are reared from AF-eggs and mated uncontrolled, you may profit from heterosis effects (in workers) in each generation, but at the same time keep ‘your’ race/breeding line/etc. ‘pure’. But one can think of many more applications. Compared to AM, time schedules are reduced significantly.
Except from the syringe and the glass capillary, you do not need any special equipment. For sterilisation you can use a high pressure cooking pot (about 120 degrees Celsius, 20 min).
1. Preparation of drones/semen:
Let the queen lay unfertilised eggs in drone combs about 40 days (24 days development + ca. 16 days for sexual maturity)before AF. These drones should hatch under controlled conditions to be sure about their origin. If e.g. marked with colour paint on their thorax, they can be allowed to freely fly because only a few are needed (one?).
Here comes the most difficult part:
Fill a sterile syringe (volume about 1 ml) about half with sterile sperm dilution buffer (same as used as a stop solution in AM). Attach a sterile glass capillary (about same as with AM) to the syringe and make the dilution buffer fill the capillary, then draw back to make a small volume of air enter the capillary. This air bubble is used to separate the semen from the dilution buffer (this large portion in the ‘back’ of the syringe). The semen of one selected drone (about 1 ul) is then collected in the glass capillary (same procedure as with AM). Thereafter, draw about eight to ten times the volume of the semen (i.e. 10 ul) of semen dilution buffer into the syringe and mix the semen and this small volume of buffer through repeated draw and push cycles on a sterile glass plate. The prepared, diluted semen can be used several hours if stored at room temperature and in the dark.
2. Preparation of unfertilised eggs:
In the morning, cage the queen on one side of a empty fully drawn drone comb. It takes some time till she begins egg laying. In the afternoon transfer the queen to the other side of the same comb. Now, she will continue egg laying after a few minutes. These eggs can now be used for AF. (My comment: In another article I read that if unfertilised eggs will enter development spontaneously after about 4 hours. So in practice, one could wait about 2-3 hours before taking the drone comb to ensure that enough eggs were laid).
The syringe with the diluted sperm is pushed so that the diluted sperm forms half a droplet at the end of the glass capillary. A egg to be fertilised must now be covered with diluted sperm at it’s upper 25 % (the free end of the egg not being attached to the cell)for a second. That’s it! To prevent the sperm from drying, the droplet is drawn back into the syringe each time after AF. Be sure to mark the respective cells (using e.g. an overhead transparency) on the comb.
The comb with AF-eggs is then transferred to a previously dequeened colony. After 3 days the larvae can be grafted as usual. The raised queens can be used for AM too, of course.
If queens are reared from AF-eggs and mated uncontrolled, you may profit from heterosis effects (in workers) in each generation, but at the same time keep ‘your’ race/breeding line/etc. ‘pure’. But one can think of many more applications. Compared to AM, time schedules are reduced significantly.
Except from the syringe and the glass capillary, you do not need any special equipment. For sterilisation you can use a high pressure cooking pot (about 120 degrees Celsius, 20 min).
