# Is it possible to graft too young?



## Oldtimer (Jul 4, 2010)

In theory the younger the better. BUT.. grafting really young, just emerged from the egg larvae, there may not yet be much royal jelly around them and the larvae are put into the new cell in a pretty dry condition, that can cause poor acceptance.


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## Shibukraj (Mar 13, 2017)

I concur with point of view of @Oldtimer. I had the same observation. In my quest to raise the best queens i went to the smallest of the larvae and landed in poor acceptance too. What i observed in that exercise, the amount of royal jelly is very low and it either gets the cups dry quickly or may be the larvae is getting damaged during the graft. I dont remember which thread I read, but this topic has been discussed here in bee source earlier as well. While grafting go for not too small not too big, and the one with much of royal jelly for a good acceptance rate.


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## little_john (Aug 4, 2014)

If it'll help any - this is the size of larvae recommended by Sladen, Frank Pellet etc:










Which I suspect is a tad larger than many people are selecting for ... ?
LJ


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## JWPalmer (May 1, 2017)

Thanks LJ. I am sure that many of us attempt to graft the fresh hatched larvae and find poor acceptance. I think second instar is a better size and that is what your graphic depicts. New research regarding the caste determination seems to indicate that the diet of the queen is identical to the diet of the worker bee at least in the beginning and quantity is what makes the difference. A one day old larva swimming in RJ is much easier to graft and will make a good queen IMO.


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## Steve in PA (Jan 26, 2015)

Wow, thanks LJ. The ones I normally look for are labeled too small on that picture and I pass up the ones labeled right size.

Next round in 2 weeks I will change course.


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## crab414 (Jan 6, 2020)

Morning Steve. What would happen if you primed your cups with a good amount of royal jelly? I wonder if this would help in sliding off the larve? These are the things I'm going to play with next year.


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## gone2seed (Sep 18, 2011)

Question for those of you who prime. How do you store your rJ from one grafting session to the next? I have been harvesting ,and using ,fresh but that wastes some.


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## JConnolly (Feb 21, 2015)

3 out of 18 is better than I got my first couple of tries. Five years on and I got 7 out if 16 my last round. I still suck at grafting, so I graft twice as many as I need. Don't give up.


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## BernhardHeuvel (Mar 13, 2013)

little_john said:


>


There is not a single larvae on that picture that I would graft from. All those larvae are way too big. Even the one labeled too small. 

We weigh our queens at birth and with larvae as shown in the picture you don't get queens that weigh more than 200 mg. We average 235-250 mg and the heaviest queens weigh 290 mg. (The birthweight is related to a) longevity b) egg laying capability [due to bigger size of ovaries and spermatheca] and some more parameters. 

The best advice is: you graft those larvae that you can't see. 

Because freshly hatched larvae are transparent and fresh royal jelly is crystal clear, too. So you have a glass like larvae in a glass like jelly. You see the ribbons on the surface, this is where you dig for the larvae.


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## Steve in PA (Jan 26, 2015)

I primed this round for the 1st time ever. I used diluted honey/h2o. 

This has just been a bad year for me grafting. Usually I'm in the mid to upper 80%. Maybe it's just my eyes getting older.

The bad % so far this year is why I changed it up & primed.

The clear ones Bernard describes are what I mostly had in the grafts that didn't take. I am going away for a week but will experiment when I return.


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## little_john (Aug 4, 2014)

BernhardHeuvel said:


> There is not a single larvae on that picture that I would graft from. All those larvae are way too big. Even the one labeled too small.
> 
> We weigh our queens at birth and with larvae as shown in the picture you don't get queens that weigh more than 200 mg. We average 235-250 mg and the heaviest queens weigh 290 mg. (The birthweight is related to a) longevity b) egg laying capability [due to bigger size of ovaries and spermatheca] and some more parameters.
> 
> ...





> Hundreds of experiments in using larvae from three hours old, up to those of 36 hours, prove that Queens from the former are in no way superior to those from the latter, while the bees always choose the latter, where the power of choice is left to them. As all of my plans of rearing Queens require the changing of small larvae, I have dwelt thus largely upon this very important point, so that the reader might know just where I stand in this matter. Years of success in producing the best of Queens, together with the result of many experiments, conducted by some of our best Queen-breeders, go to prove that I am correct in the above conclusion.
> *Doolittle - Scientific Queen Rearing, p.43*


Sounds like there's something of a conflict of opinion here ... 
LJ


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## calkal (Feb 2, 2019)

As each week goes by and I graft again I find that I am going for younger and younger larva. So I have been wondering about this same question is there such a thing as grafting too small of a larva?


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## msl (Sep 6, 2016)

> There is not a single larvae on that picture that I would graft from. All those larvae are way too big. Even the one labeled too small


Agreed



> Sounds like there's something of a conflict of opinion here


well it IS beekeeping :lpf:

however I will take Taber and Tarpy over Doolittle... a LOT has been learned in the last 130 years


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## psm1212 (Feb 9, 2016)

Like Bernard, I would not graft anything in the picture posted by little john due to my belief that all of them are too old. Even the one that says "too young." I am not stating that I think I am necessarily correct. While I think I have finally gotten fairly proficient and usually get good percentages of capped cells, that is not to say that I would not have had the same success with larger larvae. 

As to the resulting queens from 3 hr vs. 36 hour-old larvae, again, I don't have experience with grafting the older larvae. 

If Doolittle's observation cited by little john above continues to bear true, it would make grafting a lot easier if only that you would have so many more eligible larvae per frame to chose from.

Worth investigating more.


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## Oldtimer (Jul 4, 2010)

There is a solution to this conflict. Me, I would not graft the "right sized" one either. But in that diagram none of them appear to be swimming in royal jelly. 

Since latest research indicates the determining factor between worker and queen is quantity of food, none of those larve should be grafted from as they all appear starved, in terms of how much royal jelly we would see them in if they were in a queen cell. 

Seems like you have your breeder hive well fed, so the larvae you graft whatever the age, are swimming in royal jelly before you graft.

https://royalsocietypublishing.org/doi/10.1098/rspb.2020.0614


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## little_john (Aug 4, 2014)

msl said:


> however I will take Taber and Tarpy over Doolittle... a LOT has been learned in the last 130 years


True - a lot of stuff has been learned, but the most of the principles are as true today as they were back then - certainly the bees haven't changed one iota during that time.

ROB Manley writes that grafted larvae 12 hours or younger are less frequently accepted. Wilkinson and Brown are of the same opinion. I think this is what Doolittle was referring to when he wrote that bees prefer slightly older larvae when given a free choice.

If Doolittle's claim is indeed true, it would support ROB Manley's assertion that natural queen cells are invariably superior to artificially-created q/cells. So maybe we have learned a LOT - but perhaps not much about what's really important.
LJ


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## crofter (May 5, 2011)

I have read reports on a few experiments with variables of larvae age, dry grafting, double grafting, grafting with added royal jelly etc., and non of them considered skill of the grafter. No multi operator control. That variable can make quite a difference that could well be confusing conclusions. It sure is easier to graft 2 day larvae than ones less than 24 hours.

Aside from the physical transfer of the larvae, at least in modern experiments, the 24 hour or less larvae from a well fed comb results in a queen with superior potential. If your management system is low key that potential may go unnoticed.

The thrust of this is that 2 day old larvae may just _"fit your pistol"_ very well.


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## BernhardHeuvel (Mar 13, 2013)

Frank is right. It is often the operator of the grafting tool that damages the larvae.



Steve in PA said:


> I primed this round for the 1st time ever. I used diluted honey/h2o.


Honey and water? Why is that? For good results use pure royal jelly. 

The use of royal jelly and dipping the plastic cups into wax leads to premium results. For example, this was yesterday:




https://youtu.be/cmNk-_yszrw

Those are 24 hour old cells. I can do this repeatedly – almost all season long, which is April to September. Also I can make many more cells per starter. With a Brother Adam starter (originating from Ferdinand Gerstung, 1905) and royal jelly+wax dipping I can make 210 cells per starter. Even a standard starter makes 100+ cells with this technique. And I can do these numbers AND have birthweights of 235 mg average.




little_john said:


> Sounds like there's something of a conflict of opinion here ...


I don't run my business on opinions but on my experience, my state of knowledge (I am learning constantly something new), my observations and so on. If someone shows me another way to make premium queens, that is easier and with the same or better success, I will gratefully dump all of my old techniques.



little_john said:


> ROB Manley writes that grafted larvae 12 hours or younger are less frequently accepted.


From my observations: Bees initially prefer slightly older larvae. This is maybe due to the panic and emergency to get a new queen as fast as possible. Older larvae grow quicker into a new queen. 

BUT after 24 hours bees switch to younger larvae and rule out all the older larvae! I can say this for sure, since you can change your grafting success by this: If you use 60 % young larvae and 40 % older larvae, the bees first care for the older larvae, but after a day those larvae get chewed out. And the grafting success is 60 %. 

By caging the queen onto an empty comb five days prior to grafting, you get a full comb of larvae that are the same age (almost). By carefully grafting only the youngest larvae at the same age, grafting success is well over 95 % all season long! By using old and young larvae, you can modulate and replicate grafting success and failures. The more older larvae you use, the greater the failure. 

Don't believe it, try and observe yourself. I can only tell you, how I am successful and get premium quality queens.

But both observations can be true: bees prefer older larvae. Bees prefer younger larvae. It is only at which part of the development you look at, if the statements are true.



little_john said:


> Wilkinson and Brown are of the same opinion. I think this is what Doolittle was referring to when he wrote that bees prefer slightly older larvae when given a free choice.


As above.



little_john said:


> it would support ROB Manley's assertion that natural queen cells are invariably superior to artificially-created q/cells.


If you are not experienced enough maybe. I can say surely, that artificially raised queens are much superior when it comes to fertility, longevity and health than any natural queen. IF you know what you are doing. Using royal jelly, dipping the queen cups in wax when grafting, plus using an incubator, plus letting the queens hatch in the incubator AND feed them the first days with a mixture of royal jelly and honey every two hours, makes much better queens than any natural queens. 

I kept bees in fixed comb hives and letting them swarm naturally for 15 years now, I had more than hundred of skeps and Warré hives, and when I compare those natural queens to artificially raised queens that we produce today there is a clear winner. The artificial queen. Lives longer, makes much more brood, stronger colonies, much less diseases. I took dozens of five year old queens into honey production this Spring, and they made a comparable amount of honey compared to their younger fellow queens. I cannot see this with natural queens. 

The reason is: if you fulfil all physical needs and use good genetics on top – the results are better than nature, because there is a wide variation of fulfilment of the supplies in nature. Temperature varies, jelly varies, so you get mixed results. If that is good or bad in the long run, we all don't know. I guess, for survival alone, the variation maybe good. Although colonies with bad quality queens do not stand much stress from my experience. Nature selects for survival (and sometimes extinction...) not for performance.



little_john said:


> So maybe we have learned a LOT - but perhaps not much about what's really important.


We learned to produce healthier hives. Resulting in bigger honey harvests – even under negative developments of the environment.


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## Oldtimer (Jul 4, 2010)

Very impressive Bernhard, those are great looking cells for 24 hours. :thumbsup:


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## little_john (Aug 4, 2014)

"Better than Nature ..." Oh dear 

Seems that the Eugenics Movement (which has featured in English and American history, as well as in Germany) hasn't gone away - but moved it's focus away from humans and towards Honey-bees.
LJ


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## calkal (Feb 2, 2019)

Yes indeed Bernhard...beautiful cells!


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## kilocharlie (Dec 27, 2010)

Age of the larva, skill of the grafter, time of day, keeping them warm and moist...they are all important factors.

The most important factors I have found are: 1) grafted during the peak increase of the main nectar and pollen flow in the Spring; 2) the number of 5- to 10 day old nurse bees present. 10 imported frames of capped brood 10 days before grafting is huge, as it provides 30,000 excess nurse bees in an abnaturally strong queen cell raiser hive; 3) the colony is abnaturally fed with real pollen frames imported, pollen substitute patties, 1:1 sugar syrup, and a MAJOR nectar / pollen flow right outside the door.

So, too much food and too many nurse bees eager to feed the queen cells, suddenly made queenless at 72 hours after egg-lay and not a larvae nor an egg to feed anywhere...AAAHGGGG! Then, suddenly, at 80 to 90 hours a beekeeper gives them the grafts. Whoopie hayoo kayee!!! It suddenly becomes a SUPERCHARGED ROYAL JELLY FACTORY on steroids. The nurse bees DIVE IN and feed the grafts royal jelly like you've rarely seen.

My best acceptance rates occur when the larvae are between 80 and 90 hours after the queen mother has been isolated on the new comb in the queen excluder box known as a Pritchard box. I thought I had invented them, and I used to call them "breeder queen jails", but Pritchard came up with the exact same idea long before I did. Oldtimer pointed out that the Pritchard box helps to prevent one queen emerging too early and destroying other queen cells, as it aids in keeping the larvae all very nearly the same age.


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## little_john (Aug 4, 2014)

kilocharlie said:


> My best acceptance rates occur when [...] the queen mother has been isolated on the new comb in the queen excluder box known as a *Pritchard box*. I thought I had invented them, and I used to call them "breeder queen jails", but Pritchard came up with the exact same idea long before I did. Oldtimer pointed out that the Pritchard box helps to prevent one queen emerging too early and destroying other queen cells, as it aids in keeping the larvae all very nearly the same age.


Never heard the term "Pritchard box" before - I guess it's the same as the "Dutch Cage" Laidlaw mentioned in one of his books ? I tried for ages to get more info on this rare beast called a Dutch Cage, only to find that most people call these "Queen Traps", and that their use is fairly common.

Here's an all-metal version from Thornes here in the UK - which is not particularly cheap:










Not that difficult to make your own (although not as grand as the above) - I keep meaning to make one - 'tis on the 'to do' list ... 
LJ

PS - if making your own - it has occurred to me that if this box had interchangeable sides: QX *or* Mesh, then it could also be used as an infallible means of introducing a queen to a hostile colony. Just a thought.


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## Steve in PA (Jan 26, 2015)

I guess I never updated this thread. After having miserable results the entire season with the nicot cups no matter what I did I decided to make a change. I pulled out my trusty JZBZ frame with cups and used it for the last round of grafting of the year. No priming or any other frivolity to try and improve my take rate. 80% of them took.

I rigged up the nicot cages on the JZBZ cups using craft sticks and wire. That also worked and enabled me to keep the virgins banked until I can get the nucs ready for them. I will post pictures of that arrangement later.

So, it was reassuring to me to know that I can still graft and whatever is wrong seems limited to the nicot cups. They were knockoff so maybe the plastic has something they didn't like?


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## BernhardHeuvel (Mar 13, 2013)

Is it the real Nicot plastic or some cheap Chinese copy of Nicot?

Some distributors startet selling "Nicot" cups and it wasn't the original but the cheap stuff. Still they advertised it as "Nicot"...the more honest distributors called it "Nicot like"...


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## BernhardHeuvel (Mar 13, 2013)

little_john said:


> "Better than Nature ..." Oh dear
> 
> Seems that the Eugenics Movement


Dear, Eugenic... :s


Nature never breeds for the best – contrary to the common belief (read: misunderstanding) what Darwin said. It breeds for the average and for survival. 

That manmade queens are "better" than natural queens is not a surprise. Nothing to do with eugenics. What is better for us, is not better for Nature. The way it is. We don't eat wild carrots but garden carrots. Garden carrots are bigger and "better". For us. Not for the rest of the living World. But for us. Because it feeds us, unlike wild carrots.


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## wildbranch2007 (Dec 3, 2008)

I started grafting at 12 yrs old, I would go any younger than that. :scratch:


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## jim lyon (Feb 19, 2006)

kilocharlie said:


> Age of the larva, skill of the grafter, time of day, keeping them warm and moist...they are all important factors.
> 
> The most important factors I have found are: 1) grafted during the peak increase of the main nectar and pollen flow in the Spring; 2) the number of 5- to 10 day old nurse bees present. 10 imported frames of capped brood 10 days before grafting is huge, as it provides 30,000 excess nurse bees in an abnaturally strong queen cell raiser hive; 3) the colony is abnaturally fed with real pollen frames imported, pollen substitute patties, 1:1 sugar syrup, and a MAJOR nectar / pollen flow right outside the door.
> 
> ...


Great post. I would only take issue with the requirement of an ongoing major nectar flow and no larvae or eggs in the hive or did I misunderstand. I’ve found heavy nectar flows to be a nuisance as the bees are so anxious to build comb around the grafts. Love a heavy pollen flow with a minimal amount of new nectar and then supplement with light feed regularly. While a lack of open brood is certainly desirable, I don’t see it as a requirement if the hive is sufficiently populated. I do make a point of minimizing open brood adjacent to newly started cells however.


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## little_john (Aug 4, 2014)

BernhardHeuvel said:


> Dear, Eugenic... :s
> 
> Nature never breeds for the best – contrary to the common belief (read: misunderstanding) what Darwin said. It breeds for the average and for survival.


Agreed - but Darwin never said "best" in the sense of highest performing - he said "best fitted" (as in "survival of the fittest" - which were actually Wallace's words, not Darwin's) - meaning best suited for a particular ecological niche.



> That manmade queens are "better" than natural queens is not a surprise. Nothing to do with eugenics. *What is better for us, is not better for Nature.* The way it is. We don't eat wild carrots but garden carrots. Garden carrots are bigger and "better". For us. Not for the rest of the living World. But for us. Because it feeds us, unlike wild carrots.


That is *exactly* what was at the heart of my earlier comment. You appear to have bought into the Judeo-Christian Genesis doctrine which separates human beings from the rest of the Natural World - as if we are completely separate from it - nothing to do with us, etc. The Earth and it's wildlife - plant-life and animals - have been placed on the Earth for us to use according to our whims and fancies.

Of course Nature breeds to the average - that's* why* it's the average. So why don't we ? "More and More" - it's one of our basic paradigms. Clever, but not very wise - because a continuous increase of anything is impossible in the long term. And killing-off the genes of what we view as being lesser performers most certainly *is* Eugenics. 
LJ


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## kilocharlie (Dec 27, 2010)

Well, Jim, if the nectar flow is THAT good, yeah, wait for walnuts to bloom - mostly all green pollen. There are others, but you are Jim Lyon, and you perhaps know them better than me!!! 

Perhaps I left out that the bees should be crowded real thick in the cell builder. Boiling over the top. 

Yes, I do start the queen cells in QUEENLESS cell builder just to get absolute control of the timing so they all start at once, and they tend to start plenty of cells that way. Not that it is a must, it just helps prevent one virgin queen from emerging early. They can go back to queenright later.

**************

Little John - My Pritchard boxes are wooden on the bottom and ends, made of queen excluder on the sides, and they have a pop-off sheet metal top. I make a sheet metal angle to hang the Pritchard box on the frame shelf of the beehive box. The boxes are 3 frames and 4 bee spaces wide on the inside, and fit inside like an oversize frame or feeder. 

The special frames that go inside the Pritchard boxes are shorter and not as deep as normal frames - the Pritchard boxes have their own frame shelves. They are not as deep because the box is the depth of a frame, and they have a wooden floor, and you need to leave a gap between the Pritchard cage frame and the floor of the box.

In order to get the Pritchard box frames drawn out, I screw them onto a normal sized frame top bar with 2 drywall screws. The bars are painted pink for quick identification. I place one at a time in between two frames of open brood in a strong, increasing colony.

They are used for isolating a breeder queen so that I know where she is and don't have to look all over for correct age brood at grafting time, to be sure it is HER brood, and it helps prevent one queen emerging a day before the others and going on a queen cell killing spree.


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## little_john (Aug 4, 2014)

kilocharlie said:


> Little John - My Pritchard boxes are wooden on the bottom and ends, made of queen excluder on the sides, and they have a pop-off sheet metal top. [...] etc.


Many thanks for that description. I'd never heard the term 'Pritchard Box' before, nor heard of anyone using custom-sized frames in such a way.

This was the first reference I came across re: such cages (from Laidlaw) : 








in which he uses a cage to keep the Queen out - away from q/cells - rather like how we use roller cages today. But the beauty of these types of cage is that they can either be used to keep the Queen IN or OUT, and so could be very flexible in their use.
With mesh sides, seems to me such an all-encompassing cage could even be used to introduce a Q+ve Nuc into a working Q-ve colony 'on the fly', without any drop in activity. Maybe. Or at least a frame of emerging brood with a Queen - not unlike an over-sized push-in queen cage.
'best
LJ


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## jim lyon (Feb 19, 2006)

kilocharlie said:


> Yes, I do start the queen cells in QUEENLESS cell builder just to get absolute control of the timing so they all start at once, and they tend to start plenty of cells that way. Not that it is a must, it just helps prevent one virgin queen from emerging early. They can go back to queenright later.
> 
> .


Of course, big difference and I agree, It does give you better control of the process. Sounds like you’ve got a good system going. We use queenright cell builders and as long as we see ample rj at 24 hours, we usually allow them to finish as well. I won’t get long winded about my take on the pros and cons. 
BTW, Ive seen this only rarely and heard other cell raisers confirm that builders will occasionally “cut” cells in a heavy flow. 
Green pollen from walnuts? Interesting, is that a pro or con for cell production?


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## kilocharlie (Dec 27, 2010)

I wish I knew more. Thee walnuts here tend to bloom 2 or 3 weeks to a month and a half after I've grafted my first batch of queen cells. The walnut orchards are declining, mostly 125+ years old, and not many trees left. 

That said, my best guess is that it would help a queen rearing effort - the bees do seem to like walnut pollen, but it seems that very little nectar comes from them. If your colonies buildup rate has you raring queens coinciding with walnut bloom, I'd bet you'd do well.

When I get my apiary back up to enough colonies to go out pollinating again, I'll take a bunch up to a large walnut orchard near one of my fishing holes right as they bloom. I'll try to run a couple batches of queen cells in the 6-frame ventilated nuc's and see how that pollen flow works out.

That orchard is in it's production prime and well cared-for. I'll check with the owner to get his permission and see how he is about spraying. It's about 250+ miles North of Ojai, so the timing may be different. I suppose that star thistle may be blooming by then, and you know mustard will be open. I hope the fishing is good. I'll probably stay at the lake while I'm not working the bees.


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## jigsaw (Jun 10, 2019)

kilocharlie said:


> I hope the fishing is good. I'll probably stay at the lake while I'm not working the bees.


That's a terrible idea. *jealous*


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## Knisely (Oct 26, 2013)

BernhardHeuvel said:


> The use of royal jelly and dipping the plastic cups into wax leads to premium results.


Do you just dip the rim of the queen cup, or is the entire inside of the queen cup waxed?


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## BernhardHeuvel (Mar 13, 2013)

*Transcriptomic, Morphological, and Developmental Comparison of Adult Honey Bee Queens (Apis mellifera) Reared From Eggs or Worker Larvae of Differing Ages*

Discussion

Fertile queens can be produced by the commercial queen breeding practice of rearing queens from transplanted young worker larvae, but our research shows that adult queens of different larval rearing types (E, L1, L2, and L3) differed in their morphology, development time, and transcriptome.

Queens reared from older worker brood are lighter (Fig. 2D) and had fewer ovarioles (Fig. 2E) than queens reared from eggs. This result is consistent with earlier studies (Woyke 1971, Hatch et al. 1999, He et al. 2017) and successfully replicates an important result. The reduced ovariole number of queens reared from later age worker larvae is a concern because ovariole number is a key factor for queens reproductive capacity (Woyke 1971, Bouletreau-Merle 1978), which has a direct impact on colony growth and productivity (Rangel et al. 2012).

Our data showed that there were differences in development between our rearing groups post-transfer to the same uniform queen cells. The duration of development until emergence differed between our groups: developmental time increased significantly as the age of the transferred larva increased (Fig. 2C); therefore, queens reared from older larvae developed more slowly than the queens reared from younger brood. Typically workers have a significantly longer developmental period than queens (despite being smaller), and so this result suggests that the developmental cycle of queens reared from worker larvae is partially intercaste. The consequences of a longer developmental time would be significant in a natural colony. A colony would naturally produce several queens who, on emergence, fight to the death until only one remains. The first queen to emerge has a major advantage because she can kill her rivals, whereas they are still in their queen cells (Winston 1991). In a natural colony, queens reared from an older worker larvae would be unlikely to survive because of their lighter weight and slower development.

The queen cell lengths were shorter for queens reared from older worker larvae than queens reared from eggs (Fig. 2A and Supp Fig. S1 [online only]), and the amount of royal jelly remaining in the queen cell postemergence of the adult queen also decreased with age of transplant (Fig. 2B). We propose that the decline in residual royal jelly with increasing age of the transplanted larva could be due to workers having less time to provision the queen larva prior to pupation (Guo et al. 2015). This factor, along with the larvae obtaining a smaller size by pupation, probably contributes to the decrease in size of the sealed queen cell. The important point is that there are consequences of the age of larval transfer for the entire developmental cycle of the queen. The developmental environment did not become uniform once eggs or larvae were transferred to the artificial queen cell. This probably contributed to the differences in adult queen phenotype we observed and the differences in transcriptomes.

Number of DEGs of larvae (L1, L2, and L3) groups compared with E group increased with the transferred age of worker larvae. Further analyses showed that many of these genes had functional classifications related to reproduction, longevity (Corona and Robinson 2006, Barry and Camargo 2013, He et al. 2017, Yin et al. 2018, Walton et al. 2020), immunity (Barribeau et al. 2015, Boutin et al. 2015), or metabolism (Figs. 3 and 5; Supp Tables S4 and S6 [online only]). This is concerning because these are key traits that differentiate queens and workers and directly contribute to queen health, longevity, and vigor. If queens are compromised in these traits, it threatens the survival and growth of whole colonies.

KEGG analysis indicated that DEGs of all comparisons between larvae groups and E group were mainly involved in metabolic pathways (Fig. 4 and Supp Table S5 [online only]), while the greatest and most abundant differences of DEGs in the comparison of queens and workers were also in genes related to metabolic processes (Severson et al. 1989, Corona et al. 1999, Evans and Wheeler 1999). This suggests that queens reared from worker larvae differ in metabolic function compared with queens reared from worker eggs.

In summary, our data reinforce the interpretation that queens reared from older worker larvae are partially intercaste (Severson et al. 1989, Corona et al. 1999, Evans and Wheeler 1999). Queens reared from older worker larvae, while viable and appearing normal, had reduced reproductive capacity. Transcriptomic analyses suggested compromised immunity and longevity in queens reared from older worker larvae. In view of this, we recommend transferring eggs or larvae as young as possible from worker cells to queen cells for the rearing of high-quality queens.

Yao Yi, Yi Bo Liu, Andrew B Barron, Zhi Jiang Zeng, Transcriptomic, Morphological, and Developmental Comparison of Adult Honey Bee Queens (Apis mellifera) Reared From Eggs or Worker Larvae of Differing Ages, Journal of Economic Entomology, , toaa188, https://doi.org/10.1093/jee/toaa188

https://academic.oup.com/jee/advance-article/doi/10.1093/jee/toaa188/5899428


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## kilocharlie (Dec 27, 2010)

jigsaw said:


> That's a terrible idea. *jealous*


Yes, I hate it when the fishing is just too **** good! But then again, that has only happened to me once, when I went to Lake Casitas to find peace and quiet to study for final exams. I caught t full buckets of crappie - they just would not stop biting. I finally put the tackle down and read my physics book, my notes, and went over my homework. That was a difficult decision.


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## kilocharlie (Dec 27, 2010)

Bernard - your posts are just too awesome. I know to really perk up and memorize your information. You are a true hero of Beesource. Thank you.

Thanks also to all the other real pros who take time to explain things here - Oldtimer, Jim Lyon, Michael Palmer, JWChesnut, Fusion-power, Michael Bush, and to 40 or so others who have put so many truths and practical experience here for us all to benefit.

Thanks to all those hobbyists who have contributed excellent bits and pieces along the way, and for asking so many questions! These have helped!

Thanks also to the professors at the universities, to Randy Oliver, to Joe Traynor, to Dr. Gordon Wardell, Dr. Seely, Kim Flottum, the people at the Federal Bee Research Laboratories, and the many, many more who take beekeeping so seriously and keep moving us forward.

Thanks to those who have gone before us and paved the way to success. Their names are legendary, and their books and articles have guided our efforts, our research, and saved us countless hours of effort.

The helpful information, practical advice, strategic thinking, and wisdom have been the biggest factors in my learning this game. I owe a great debt of gratitude to all.


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## Saltybee (Feb 9, 2012)

kilocharlie,
good to have you back connected.


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## AramF (Sep 23, 2010)

Bernhard, I've watched your youtube video on raising queen, and noticed you plunge your cells into molten wax 3-4 times. Are you doing this to save the bees the effort of making wax? Most of the time bees reuse wax from frames to make queen cells, that's why they are always the color of the frame. Maybe you are doing it for sanitary reasons, or earlier deep cells allow for more RJ storage?
Also, do you allow bees to clean/polish the cells before grafting into them?

Aram


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## Steve in PA (Jan 26, 2015)

Wow, this is an old thread I forgot about. Problem was something with the cups. I didn't throw them away and on a whim I tried using a few this year. Whatever smell or chemical they didn't like must have been gone because I got back to normal acceptance. I did switch over to a sable brush as my primary tool now so maybe that had something to do with it too?


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