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Discussion Starter · #1 · (Edited)
This is being developed. It is like the liquid nitrogen test, but easier because it is a spray. It could make hygienic breeding much easier. It seems like they uncap then recap the brood without killing it.
 

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Right now ubo isn't available to just buy right now in the form of some spray. Word on the street though is that it's in development. It would indeed make assay test much easier and quicker. Not only can you do the nitrogen test but some breeders will uncap purple eye pupae and count reproductive mites. Look up harbo assay for more info on that vsh test.
 

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Below is a clip from a post [email protected] A method to see if your bees are active uncapper/recappers. Having a spray or paint on solution to cover a marked area of capped brood and see what the bees do with it, would certainly be simpler.

{While certainly more 'quick-and-dirty' than the precise protocol you have developed, Dr. Seeley sent me this idea, which I thought was pretty good:

"I do not know if there is a standard protocol for this assay, except that researchers like to sample ca. 100 capped cells from the frames of capped brood in a colony. This can be done by pressing a patch or two of duct tape down onto a frame of capped brood in a colony, then pulling it up and studying the undersides of the cells’ cappings. By sampling 100+ cell cappings, one can get a good estimate of the % of cells that have been capped and recapped.

A cell capping that has been capped and recapped is recognized as follows: "The recapping behaviour can be easily detected as a hole in the spun cocoon of the pupated larva ranging in size from one mm to the entire area of the cap. The hole is subsequently covered over with wax by the adult bees. This hole can be seen as a dark, matte spot on the underside of the cell cap distinct from the glossy coating of the cocoon.”

This is a quote from the paper that I have attached. Fig. 1 in this paper contains excellent photos that show more clearly than the above words about how to recognized cells of mite infested brood that have been uncapped and recapped. I hope you find that your bees are controlling the Varroa mites in this way."}
 

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A method to see if your bees are active uncapper/recappers.
Another thing that works really well is waxing strips. They have the built in advantage of being consistently sized so you pull off approximately the same number of caps every time:

Regarding this assay I was recently in an e-mail exchange with Dr. Büchler and I asked him about the specific product they employ for uncapping/recapping assays. He indicated that after trying several options, they settled on the Isana brand depilatory strips as the best solution.

It does not appear that we have the Isana brand strips available domestically, but it looks like Rossmann also makes the Veet brand.

A search for 'Veet Cold Wax Strips' produces a lot of results.
 

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Discussion Starter · #5 ·
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Reactions: Kaira Wagoner

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I should say something.
I've been given permission to use the UBO assay. Last summer we started with 64 colonies that had performed well over the previous few years. Wintering, disease's present, temper, honey produced, frozen brood assay, etc...just typical categories in a breeding program that I've been running since 1998. We chose the 5 highest scorers in all categories to be the breeders for 2022.
In 2022, we raised 100 queens...50 from the program queens and 50 from 5 breeders of my own choice as controls. So, two apiaries of 50 nucleus each...separated by 20 miles. 5 nucs from each program queen and 5 controls from my own stock selection. This summer we added the UBO assay. Understand that UBO assay and frozen brood assay are measuring different things. Frozen brood assay is selecting for brood death. UBO is selecting for unhealthy brood...brood with high varroa population or brood with low varroa population but high virus titre. I'm working with the UVM bee lab and some grant money. Took two days with two lab techs and 4 of us to do the UBO sampling. The variation in uncapping was something to see.
Next season, we continue. Selecting the best from 2022 sampling to raise the next generations of queens.
Very excited about this program. Pretty much what will keep me in bees.

Font Circle Handwriting Pattern Soil
 

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Mike, have you or someone else tried taking colonies with excellent uncapping percentage from the UBO spray and performing Harbo assays to see how they rank? ( I'll email Kaira Wagoner on this, too...)

It seems that there are two ways to determine Varroa Sensitive Hygiene behavior, and if that is the case, your top UBO-performing queens should rank as 4's on the Harbo scale.

Concerning recapping behavior:
I find myself wondering if hygienic colonies that rank high on freeze assays but low on Harbo or USO assays would show any recapping behavior. If NOT, then this might show that recapping is a good indicator of VSH traits. If they DO, then recapping may be an indicator for a wide variety of traits, which might not be as useful as we hope. (I understand that hygenic stock typically do well spotting disease, while VSH do better spotting mites. Although disease-detection is great, I'd rather focus on getting the elephant out of the hive before the mice.)

It would be wonderful if checking for uncapping/recapping behavior would give insight on VSH status of colony. What a great quick tool that would be for queen breeders to have out in the field, with minimal time and cost. Not to replace the Harbo or UBO assays, but to supplement them.

Thoughts?
 

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Mike, have you or someone else tried taking colonies with excellent uncapping percentage from the UBO spray and performing Harbo assays to see how they rank? ( I'll email Kaira Wagoner on this, too...)

It seems that there are two ways to determine Varroa Sensitive Hygiene behavior, and if that is the case, your top UBO-performing queens should rank as 4's on the Harbo scale.

Concerning recapping behavior:
I find myself wondering if hygienic colonies that rank high on freeze assays but low on Harbo or USO assays would show any recapping behavior. If NOT, then this might show that recapping is a good indicator of VSH traits. If they DO, then recapping may be an indicator for a wide variety of traits, which might not be as useful as we hope. (I understand that hygenic stock typically do well spotting disease, while VSH do better spotting mites. Although disease-detection is great, I'd rather focus on getting the elephant out of the hive before the mice.)

It would be wonderful if checking for uncapping/recapping behavior would give insight on VSH status of colony. What a great quick tool that would be for queen breeders to have out in the field, with minimal time and cost. Not to replace the Harbo or UBO assays, but to supplement them.

Thoughts?
Hi CiCi,
Great question about recapping. We have clear evidence that UBO is correlated with pin-killed and FKB, and growing evidence that it is correlated with mite non-reproduction. We are already making plans to look next into the relationship between UBO and recapping, and should know more after the 2023 field season. Despite being correlated with other hygienic parameters, though, the UBO assay is measuring something different, as mentioned by Michael Palmer above. From my experiments, including a few new ones focused on antennae that I have not published or talked about publicly yet, it looks like colony UBO response is a reliable measure of nurse bee sensitivity to and memory of specific odors associated with mite-parasitized or otherwise unhealthy (but living) brood. Our data suggests UBO high colonies can detect a range of sublethal issues such as Varroa and high virus loads (including viruses NOT transmitted by Varroa). Folks above are correct that we are working to bring it to market, with launch in the US projected for spring of 2024. To help get a sense of where interest in UBO lies and how many tests we will need to produce, we are accepting pre-orders now, with priority given at launch to those who placed a pre-order. If you're interested, you can check out our new website opterabees.com (still under construction so please forgive updates and pages still in progress!) Thanks for your interest, we are eager to get UBOs out into the hands of breeders and beekeepers so they can try it out themselves!
 

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Hi CiCi,
Great question about recapping. We have clear evidence that UBO is correlated with pin-killed and FKB, and growing evidence that it is correlated with mite non-reproduction. We are already making plans to look next into the relationship between UBO and recapping, and should know more after the 2023 field season. Despite being correlated with other hygienic parameters, though, the UBO assay is measuring something different, as mentioned by Michael Palmer above. From my experiments, including a few new ones focused on antennae that I have not published or talked about publicly yet, it looks like colony UBO response is a reliable measure of nurse bee sensitivity to and memory of specific odors associated with mite-parasitized or otherwise unhealthy (but living) brood. Our data suggests UBO high colonies can detect a range of sublethal issues such as Varroa and high virus loads (including viruses NOT transmitted by Varroa). Folks above are correct that we are working to bring it to market, with launch in the US projected for spring of 2024. To help get a sense of where interest in UBO lies and how many tests we will need to produce, we are accepting pre-orders now, with priority given at launch to those who placed a pre-order. If you're interested, you can check out our new website opterabees.com (still under construction so please forgive updates and pages still in progress!) Thanks for your interest, we are eager to get UBOs out into the hands of breeders and beekeepers so they can try it out themselves!
Thank you for joining the discussion! I've seen a few of your presentations to local bee clubs. Do you have a Youtube channel or another avenue for people to watch your presentations?
 

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Thank you for joining the discussion! I've seen a few of your presentations to local bee clubs. Do you have a Youtube channel or another avenue for people to watch your presentations?
Hi Ruth,
Thank you for your interest! I don't have a youtube channel but we do plan to start sending out some project updates soon (you can email us through the website opterabees.com if you want to be added to the list to receive these) and the UBO work has been featured on Inside the Hive TV here:
and here:
, with another update planned for this December (date TBD).
 

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Thank you for your response, Kaira... looking forward to this summer's research results!

Crofter or Litsinger, could you share the discussion link you refer to in the third & forth response of this discussion?

I've been digging on the web, and I want to personally thank Litsinger for mentioning Dr. Büchler's recapping studies... however, what I could find out there was incomplete. The most current item of interest in the recapping (SMR) phenomenon is this link that provides recapping assay details in Sept 2017: NEW SMR Protocol – RNSBB Unless I misread, it looked like that assay protocol was for a study being launched, but I could not find results of that study.

If anyone could point me to more recent (after 2017) discussions or studies done specifically with recapping and Dr. Buchler, I would appreciate it. I have found only general studies, like the 2022 Jan ABJ article about study of commercial beeks using mite-resistant stock in Europe, but not specific studies on recapping.

For folks needing an explanation on the Harbo assay, he kindly provides details here: harbobeeco - Measure VSH The amount of detail says a lot to this gentleman's dedication to sharing with colleagues, instead of keeping 'trade secrets' that helps only the insiders. Thank you, Dr. Harbo.
 

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Thank you for your response, Kaira... looking forward to this summer's research results!

Crofter or Litsinger, could you share the discussion link you refer to in the third & forth response of this discussion?

I've been digging on the web, and I want to personally thank Litsinger for mentioning Dr. Büchler's recapping studies... however, what I could find out there was incomplete. The most current item of interest in the recapping (SMR) phenomenon is this link that provides recapping assay details in Sept 2017: NEW SMR Protocol – RNSBB Unless I misread, it looked like that assay protocol was for a study being launched, but I could not find results of that study.

If anyone could point me to more recent (after 2017) discussions or studies done specifically with recapping and Dr. Buchler, I would appreciate it. I have found only general studies, like the 2022 Jan ABJ article about study of commercial beeks using mite-resistant stock in Europe, but not specific studies on recapping.

For folks needing an explanation on the Harbo assay, he kindly provides details here: harbobeeco - Measure VSH The amount of detail says a lot to this gentleman's dedication to sharing with colleagues, instead of keeping 'trade secrets' that helps only the insiders. Thank you, Dr. Harbo.
Here is a search link to a number of mentions by @Litsinger of uncapping recapping. What you are looking for might be in there.

 

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Here is a search link to a number of mentions by @Litsinger of uncapping recapping. What you are looking for might be in there.
Thanks for the assist, @crofter. I sincerely appreciate it!

All the information I have read concerning uncapping/recapping involving Dr. Buchler has been by personal correspondence.

That said, if you are interested in SMR/VSH here is a great recent study if you haven't seen it already:

 

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Eureka! That's the study that was eluding me... I knew the Sept 2017: NEW SMR Protocol – RNSBB had to have resulted in some kind of paper. I have some Black Friday reading to do... looking forward to it.

Thank you, Crofter, for helping me dice out search parameters for Beesource. I've searched this awesome forum for years during frequent information-seeking binges, but hadn't thought to link a user name with keywords. I can think of several other possible combos I'll need to check out while I'm playing 'catch-up' on this topic.
 
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