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  1. #41
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    Default Re: My ghetto II station

    Quote Originally Posted by beepro View Post
    Now I know what not to do next time. Either 1 or 2 time II and that's it. Whoever want to
    repeat this failed experiment can do so. I for one will not do it again.

    Not quite sure what you're saying here.

    For the CO2, test the flow rate by placing the tube in a glass of water, that way you can observe the flow rate. Without looking back at my notes, a bubble per second is roughly about right. I've never been to precise on this rate, but I've never chilled a queen either. If you're chilling queens, then I suspect that your flow rate is way too high.

    Don't expect perfection right out of the box, particularly using non-standard equipment. It may take lots of queens before you become proficient with the procedure. At Sue's class, they provided a LOT of practice queens with ideal lab conditions too, and still I had many failures.
    Horseshoe Point Honey -- http://localvahoney.com/

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  3. #42
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    Default Re: My ghetto II station

    Quote Originally Posted by AstroBee View Post
    Not quite sure what you're saying here.

    For the CO2, test the flow rate by placing the tube in a glass of water, that way you can observe the flow rate. Without looking back at my notes, a bubble per second is roughly about right. I've never been to precise on this rate, but I've never chilled a queen either. If you're chilling queens, then I suspect that your flow rate is way too high.

    Don't expect perfection right out of the box, particularly using non-standard equipment. It may take lots of queens before you become proficient with the procedure. At Sue's class, they provided a LOT of practice queens with ideal lab conditions too, and still I had many failures.
    1 to 2 bubble second is correct, i think i run a little higher than most at 2p/s, but I want her out fast as I want to get in and out fast.

    Same thing at Sue's class, I did rougly 20-30. Only the last one did we wait any period of time to check spermatheca, rest we only waited a day or so to check them. Last one came out great.
    Instrumental Insemination & Northern VSH Queens

  4. #43
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    Default Re: My ghetto II station

    Quote Originally Posted by beepro View Post
    Will leave off the gentamicin until I can get the dosage right. The 1st II queen without antibiotic is still laying. The
    2nd one died last night after the 3rd time. The 3rd virgin will be II tonight without the antibiotic depending on how it goes.
    You may want to consider leaving the antibiotic out permanently. I used it initially, but dropped it after the first year and saw no difference in the results. I achieved multi-year survival with excellent laying up until the end. Keep your conditions as sterile as possible and liberal use of saline. Clean the tips and syringe very well after use.
    Horseshoe Point Honey -- http://localvahoney.com/

  5. #44
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    Default Re: My ghetto II station

    Thanks all, for all the good encouragement. I have so many pics to document the entire II process including a complete revamping of my station set up. Every thing got elevated to see the queen better. This will make the II much more smoother when lowering the syringe. Ergonomic is very important considering I can sit there for hours on the entire process. Leaning and refining my process down. Cannot do it free style without suspending the syringe because of my shaky hand when nervous approaching the II site. Imagine jabbing extra holes on the queen when not needed.
    Since combining the 2 nuc hives together, the 1st queen is laying strong. Seems like every time I inspected the hive she is there laying right in front of me. The genetics is so stable that they are the most calm and gentle bees around. Definitely showing very good sign of the allogrooming activity in and outside of the hive. With the 4 expensive bought queens all dead only with II will continue the Cordovan lineage. Any mixing with the local carnis drones the genetics will be diluted.
    Did the 2nd round of II on the second queen tonight. After the first round last night, she's noticeably more plump today. Found her bury under a pile of bees trying to protect her. At 11 day post emergence, they already recognized her as the queen. This time I only use 14 drones instead of 30 to extract 20 ul from them and have almost 9 ul left over after the II. This queen only use 12 ul or so which is more than enough to satisfy her. I'm still using the 90 degree angle to do the II. Maybe one day I will experiment on the 45 degree when getting more proficient at it with improved II equipment too. Found out that an II virgin will not likely to leave for her mating flights when she's all filled up with more than the recommended 10 ul. There is no time for her to explore her surrounding with a QE on and the consecutive days of II on her at the right time. That is why neither of the 2 queens go on their mating flights afterward even though the QE was off after the 1st II. Because this experiment is such a success, I decided to use 20 ul from now on because not all will go in as seen from the pics. There is no harm to the queen as the extra will just leak out. At least some will go in to fill up the spermatheca. Also spent an hour or so to make 2 cap. tubes after I broke the first one on accident yesterday while cleaning it. These are very fine pointed and fragile glass tubes. The one I made only 2 out of 17 tube was a successful pull with a very fine point at the end and enough to take in 30 ul at one time. With 2 tubes of 400 total I still have plenty to work on. Seems like a longer tube is easier to work with. This is the end of my II session unless the new queen will lay me some drone eggs later on. Put in 2 drone comb frame in there hoping that she will lay some. If she does then my II experiment will continue and with some improvement too.




    Are these queen pics look familiar to you?:
    Attached Images Attached Images
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  6. #45
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    Default Re: My ghetto II station

    More pics! Intro. the queen back after the II:
    Attached Images Attached Images
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  7. #46
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    Default Re: My ghetto II station

    Quote Originally Posted by beepro View Post

    Did the 2nd round of II on the second queen tonight. After the first round last night, she's noticeably more plump today.
    Are you inseminating twice on two separate days? If so, you should probably drop back to a single insemination. I perform a single insemination, but with two CO2 doses on consecutive days. During the first CO2 treatment, she gets a wind clipped while under (you should seriously consider doing this too), and the following day she is inseminated with 10 - 12 ul and if all goes well gets a numbered disc (you should definitely at least mark your II queens). Clipping is to prevent her from flying and mating. The whole point in doing II is controlling the crosses, and you insure this by preventing the flights. Sue Cobey told us that a single insemination is just as good as two separate inseminations. In fact, one may ultimately prove more efficient since there will be less possibility of failure because of the fewer times she's handled.

    Also, the vertical technique seems to hyper-extend her abdomen (at least from your pictures). You can do that with the 45 deg setup too, but probably less likely to happen. The motion you're trying to achieve is not all axial extension, but some axial and some transverse. Regardless, VERY little extension (in either direction) is needed. It is mostly a stabilization (with some small extension) of the abdomen that you're trying to achieve.
    Horseshoe Point Honey -- http://localvahoney.com/

  8. #47
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    Default Re: My ghetto II station

    Yes, the first day on CO2 then follow by 2 consecutive day of II on the same queen. The reason being my aim is a bit
    off thinking that there isn't much done on the first round. The first queen was just a lucky shot now that she's laying. And not clipping her wing to allow her on a mating flight is to test out what drones are out there at this time. Also to see if she will go on her flights and if successful, what are the workers look like at each hatch. Are they a mixture of the Cordovan and carnis? Or they are the LIFO or FIFO of workers since someone claimed that they are a mixture of both. I think they are the LIFO of workers though which will be the carnis. But so far it seems that I've over saturated the queen not enabling her to go on her mating flights. That is o.k. since the Cordovan genetics have been preserve as well. Either way it is fine with me. If she lay the carnis then it will be a production queen the next Spring. If not then will pick her as the breeder queen.
    What is more interesting is that approaching our mild winter time my hives can still make the virgin queens. If I can somehow make the queen lay the drones to overwinter then I can still continue with this process. Some small nuc hives will overwinter their drones as I have seen before. This may turn out to be a year round II adventure. There are so many variable I have not test yet. Once the perfect II angle has been achieved and bypassing the valve fold is not an issue anymore, I will go back to a single dose with 20 ul. Almost one month into this process and I'm still learning and experimenting. So far I know what cap. tube tip size to make. This cannot be done by asking question alone as it is a hands on operation. Knowledge will only take you so far that you still have to do it in order to learn.



    1st queen is still plump:
    Attached Images Attached Images

  9. #48
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    Default Re: My ghetto II station

    Just a quick hive check tonight. Saw the recent II queen hiding among the bee cluster again. There are still many mature
    Cordovan drones left. What a waste. She's acting like a mated queen already looking into the empty cells like a real mated queen does. So maybe that my II did work the first time. But her abdomen is still too small to tell. Like the first II queen her abdomen will not swell up until she is laying for a few days. Then she will be more plump when others are feeding her more RJ. While this queen is roaming on the empty cells I
    did a combine with a weaker nuc hive that is queen less now. This should give them an instant population boost if they
    liked this queen. I will know for sure in another 5 day if this II is a successful one by 9/18. With II I'm able to cut down her
    laying time by almost 2 week instead of the normal on month time frame. Not that bad either. So you think I did it right this time?



    Still a virgin queen?:
    Attached Images Attached Images
    Last edited by beepro; 09-13-2017 at 03:39 AM.
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  10. #49
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    Default Re: My ghetto II station

    Hi, All!


    So I did a hive inspection today. Found some cap worker broods from the first II queen. It is a good sign! Have
    confidence that these are the Cordovan workers since the queen never take her mating flights even though the QE is
    off after the 1st day. I inspected the nuc hive every night for the entire duration of the II process. This queen is the light yellow Cordovan color that I'm looking to propagate. Also found her roaming on the drone frame right in the middle of the brood nest that I put in last week. Upon inspection into the drone cells I found some eggs scattered through out the frame. Don't expect perfection the first time around. And saw the queen laid in one of the drone cells too. It is my first time seeing a queen tucking her head all the way down into a drone cell inspecting it before depositing her egg. So I took out my cam and snap a few pics.
    Since we have the drone eggs, if the weather holds then in mid-October I can do another round of the II process with more graft virgin queens either from the first queen or from the 3rd II queen if she lays. We also have the half-carnis/half Italian queen mated with the local carnis drones to experiment on. This time I will try the 45 degree on the II station to see if there can be better insemination improvement. Pushing the queen rearing season to the max, do you think it is possible to make II queens all the way to December?
    Attached Images Attached Images
    Last edited by beepro; 09-14-2017 at 06:49 PM.
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  11. #50
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    Default Re: My ghetto II station

    A quick update:


    I'm very happy today! Finally on the 5th day of waiting, seems like forever, I saw an egg laid by the 3rd II queen this afternoon. So I gave them a pint jar of honey water. Last time I re-II the 2nd queen on the 4th days causing her death. This time around I learn that being patience enough is everything. I did check the nuc hive a few times but cannot find any eggs in the last few days. So like the 1st queen I have to wait 5 days in order to see some eggs after the II process. During this time it is better to wait and check for sign of any mating flights she might be doing. Of the 5 days that I look after this queen she did not go to her mating flights retaining all the II value of the Cordovan genetics. So even though the II process doesn't go as smooth as I like it to be with 20 ul of semen used and some leaked out, it is still a successful one on both occasion. Might have been the 3rd time if the 2nd queen did not died. This means that now I have 2 breeder queens for next year if everything holds through this winter. Interesting how a virgin behaved after the first II done.
    The II station have some improvement now with an adjustable know to control the lowering of the II syringe at 90 degree still. There are still many ways to improve on this station which I will adjust accordingly.
    Last edited by beepro; 09-16-2017 at 04:06 PM.
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  12. #51
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    Default Re: My ghetto II station

    It's actually pretty amazing the transformation a queens body goes through after being inseminated. For her mating flight she is trimmed down to a lean, fit, flying machine. Once she is inseminated there is a massive transformation in her abdomen with oviaries plumping up and getting ready for the job of turning food into around in season, one thousand and five hundred eggs daily. That her body can make this transformation in just 5 days is a remarkable thing in itself.
    "Every viewpoint, is a view from a point." - Solomon Parker

  13. #52
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    Default Re: My ghetto II station

    Quote Originally Posted by JRG13 View Post
    My only question is how do you know II was successful, you said you were going to let her go on a mating flight or she wasn't restricted from doing so after II.
    Today is the bees emergence day on SC (small cells) from the 1st II breeder queen. Finally I have waited for that long now.
    Yep, all the newly emerged worker bees are the Cordovan color, alright.
    So when I over II the queen she was too fat to make her next mating flight. No
    choice but to stay in until after the 3-5 days before laying. In order for a queen
    to take her first mating flight, she has to explored her outside hive surrounding first like a mini orientation flight. These queens never got that chance to explore theirs when confined inside a QE from emergence until II time. Anyways, using that much II will bogged down the queen making her unwillingly to take her mating flight anymore. The tiny bits of spermatheca stored sperm after the II, when successful, is enough to make her full and stay in to lay. I'm sure she knows when it is full just like a pregnant woman. This was a test to see if after II the queen will fly to mate or not. Now I know after this little bee experiment. It is that easy!

    Saw lots of cap drones today too. Enough on a frame for the next test of II queens, same color. Need to get the
    timing right for making the QCs this time.



    Same yellow emerged workers:
    Attached Images Attached Images
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  14. #53
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    Default Re: My ghetto II station

    Looks ok, but I hope you realize you're kind of inbreeding them very rapidly with your limited donor pool of queens and drones?

  15. #54
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    Default Re: My ghetto II station

    For now it is. I'm starting to source compatible bees for the II process. Can also open mate the daughters with the
    local carnis and cross back to the Cordovan drones again via II. Reading said that after the 2nd generation the
    bees will revert back to the Cordovan color. As long as I have a Cordovan breeder everything
    should be fine. After a few generation, the Cordovan genetics will be back to normal again. My experience is 4 generation to revert back. Not sure about the Cordovan until I have try them out. So it will be II to maintain the Cordovan line and a mix for the production queens with open mating. I'm sure 100 II Cordovan breeders will maintain the genetics here somewhat. I'll be spreading the little nuc hives through out the area to ensure the bees are there.
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  16. #55
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    Default Re: My ghetto II station

    Hi, All!

    Update: II a DL Cordovan queen today.


    In many aspect of beekeeping many will tell me that it cannot be done. Or it might
    not work using my way just because they tend to follow the norm. I like to experiment a little after
    being more proficient at beekeeping. So today I re-II a DL (drone layer) Cordovan queen for the
    first time. So they said that it cannot be done either.

    After a little improvement made on my II station, I figured that it was time to give it a little trial run. Now the II
    station has smoother syringe adjustment lever and better II hooks. I was a bit afraid that the queen might die when she's a laying queen already even though they are the drones. Maybe this was the reason that other beekeepers said it cannot be done? I know for a fact that when I was keeping tropical fish when a different color male mated with the laying female, the result is a different color offspring from the most recent male. So why can't a DL queen be II again assuming that her spermatheca has not been full of semen yet? If this queen is still alive after 48 hours then
    this will be a successful re-II done. Of course, she has to lay the normal worker eggs in order to count. If not then I will devise a better method of re-II another DL queen in the future. This way a good laying queen is not wasted-- squish! I can see that her abdomen is a lot larger (ballooned) right after the re-II process. If it cannot be done, why not?



    Cordovan queen re-II:
    Attached Images Attached Images
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  17. #56
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    Default Re: My ghetto II station

    Quote Originally Posted by beepro View Post
    If it cannot be done, why not?
    No reason, as it can be done.

  18. #57
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    Default Re: My ghetto II station

    So can I re-do the II the 2nd time after the 48 hours?
    She's not laying now since her abdomen is till fat up. Maybe she needed more time to
    prepare the semen to migrate to the spermatheca?
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  19. #58
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    Default Re: My ghetto II station

    Quote Originally Posted by beekuk View Post
    No reason, as it can be done.
    Oh well, this re-II of the DL Cordovan queen had failed miserably. Yesterday supposed
    to be the 48 hours time frame but I did not see the queen in there. Check again today and
    she's not in there either. So I did a combine with another laying queen, a Fat Bee Man's queen.
    Let's see how an east coast bee can survive on our dry and hot summer west coast condition.

    Not sure why the queen died. Might be that while doing the shut down laying period her
    body chemistry changed again therefore the workers cannot recognized her and started the
    balling process. Anyways, next time I will caged the queen after the re-II to see it past
    the 48 hours. The good news is that there isn't a mite inside that I can find. With a
    low mite level like that now I'm sure the coming Spring build up will not be affected. Let's
    see if this tf queen will hold too.
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  20. #59
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    Default Re: My ghetto II station

    Quote Originally Posted by beekuk View Post
    No reason, as it can be done.
    Oh well, this re-II of the DL Cordovan queen had failed miserably. Yesterday supposed
    to be the 48 hours time frame but I did not see the queen in there. Check again today and
    she's not in there either. So I did a combine with another laying queen, a Fat Bee Man's queen.
    Let's see how an east coast bee can survive on our dry and hot summer west coast condition.

    Not sure why the queen died. Might be that while doing the shut down laying period her
    body chemistry changed again therefore the workers cannot recognized her and started the
    balling process. Anyways, next time I will caged the queen after the re-II to see it past
    the 48 hours. The good news is that there isn't a mite inside that I can find. With a
    low mite level like that now I'm sure the coming Spring build up will not be affected. Let's
    see if this tf queen will hold too.
    Don't mix foreign bees into a virgin hive. She might get balled 100% of the time! When will you ever learn, huh?

  21. #60
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    Default Re: My ghetto II station

    Quote Originally Posted by beepro View Post
    Oh well, this re-II of the DL Cordovan queen had failed miserably. Yesterday supposed
    to be the 48 hours time frame but I did not see the queen in there. Check again today and
    she's not in there either. So I did a combine with another laying queen, a Fat Bee Man's queen.
    Let's see how an east coast bee can survive on our dry and hot summer west coast condition.

    Not sure why the queen died. Might be that while doing the shut down laying period her
    body chemistry changed again therefore the workers cannot recognized her and started the
    balling process. Anyways, next time I will caged the queen after the re-II to see it past
    the 48 hours. The good news is that there isn't a mite inside that I can find. With a
    low mite level like that now I'm sure the coming Spring build up will not be affected. Let's
    see if this tf queen will hold too.
    Died probably due to stress. Virgins are much more able to handle stress. Non virgins can't be reinseminated after the initial mating period. Too much stress, and I doubt her body is even able to asorb more semen at that point.

    The workers also probably notice something "wrong" and dispatch her anyhow.
    Instrumental Insemination & Northern VSH Queens

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