# Randy Oliver's lastest article on mite resistant queens



## squarepeg (Jul 9, 2010)

i've only had time to give it a quick once over, but as i recall there was a lot of meat and potatoes in it, especially with regard to what randy describes as his successes and failures to date:

"I’ve got nothing to hide as I walk the walk myself in attempting to breed mite resistance into my stock, and perhaps by relating what I’ve done, I might save others from repeating my mistakes, as well as sharing what I’m trying now, and, more importantly, the biological reasons for doing so."

randy oliver, abj may 2017 vol. 157 no. 5 p. 509

programming note,

this is the latest in a series of articles some of which have been discussed here:

http://www.beesource.com/forums/showthread.php?332799-randy-oliver-article-in-january-17-abj

the whole series, "the varroa problem", was started last november and is available for free viewing on randy's website, however the latest installment dr. cryberg is referencing has not been posted there yet.

http://scientificbeekeeping.com/articles-by-publication-date/

i hope to give the may article a more careful read this weekend richard and i'll be back with more comments.


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## Ian (Jan 16, 2003)

Meat and potatoes, I like it


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## Fusion_power (Jan 14, 2005)

Randy notes that he is trying to avoid doing a hard Bond test which has serious financial side effects. He does not fully understand the effects of inbreeding based on his comments. Bees can survive just fine with as few as 5 variants of the sex allele in the breeding population as shown by the bees on Kangaroo Island. What he is missing is a way to test for sex alleles and see what each queen carries. With that information, he could make informed breeding decisions that avoid the problem with duplicated alleles.

IMO, progress can be made faster with a hard Bond approach. It would cost about $400,000 to do a hard bond on 500 colonies based on each colony being worth $400 (net of expenses) of pollination and honey production per year over 2 years. By the third year, hard Bond should produce reliable mite resistance. He can achieve similar results with mite counts if he is willing to count the mites in all his colonies and breed only from the hives with zero counts. With 2500 colonies, he should find about 10 that are worth breeding from based on random gene variation.

Of note in the article, he put a mite resistant queen in an apiary with susceptible and heavily infested colonies. The result was predictable. Horizontal transfer brought enough mites into the resistant colony to make it appear susceptible.


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## psm1212 (Feb 9, 2016)

Can't wait to read it. I have been following Randy's series of articles. I think Randy has been keeping it at a 30,000 foot view so far. Sounds like he dives into the weeds on this one. It intimidates me that Cryberg needed to read the latest article 3 times to absorb it. That probably leaves little hope for my understanding. Anyway, looking forward to reading it. Hopefully this thread will get hot and help me digest it.


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## Richard Cryberg (May 24, 2013)

Fusion_power said:


> He does not fully understand the effects of inbreeding based on his comments. Bees can survive just fine with as few as 5 variants of the sex allele in the breeding population


I do not think you understand it as well as Randy understands it. After all, it only takes three alleles to make a queen that never lays a single diploid drone.


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## Fusion_power (Jan 14, 2005)

> I do not think you understand it as well as Randy understands it.


 Perhaps true. Though give me the due that I spent several months studying genetics and bee genetics in particular. It is certainly possible that I might know a bit more than most.

Consider a mating where a queen is raised and AI mated with her own brothers, i.e. drones produced by her own mother. What percent of the AI queen's eggs will hatch into normal females? Consider only the effect of the sex allele and allow for her to be mated with semen from say 20 drones.


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## Richard Cryberg (May 24, 2013)

Fusion_power said:


> Perhaps true. Though give me the due that I spent several months studying genetics and bee genetics in particular. It is certainly possible that I might know a bit more than most.
> 
> Consider a mating where a queen is raised and AI mated with her own brothers, i.e. drones produced by her own mother. What percent of the AI queen's eggs will hatch into normal females? Consider only the effect of the sex allele and allow for her to be mated with semen from say 20 drones.


There is no single right answer. The best that can be done is give a range. From 50% of the offspring up to 100% will be normal workers and the rest diploid drones.


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## Fusion_power (Jan 14, 2005)

Given the conditions, there are two correct answers. Under normal conditions, the daughter queen will have 1 allele in common with her mother and one from a drone. If the drone allele is different - which should be true most of the time - then the mating will result in 75% brood viability. If the drone allele is identical with one that the mother queen carries then the fact that a queen was produced automatically means the daughter queen has the same two sex alleles the mother queen carries. In this rare case, brood viability will be 50%. No matter the amount of inbreeding, it is not possible to get below 50% brood viability based on the sex allele.

So the question to ask is, how much duplication at the sex allele locus can honeybees take before colony survival is compromised? There are some decent articles covering this topic. The problem Randy Oliver will encounter over time is that he will have concentrated two sex alleles at a frequency that could approach 25% each in his breeding population. This is potentially disastrous for long term breeding, but with appropriate measures can be countered by selecting colonies based on brood viability and by maintaining very good pedigree records. A much better solution would be to develop a genetic test to determine which sex alleles each queen carries and select queens to produce drones such that duplicate alleles never occur.


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## bucksbees (May 19, 2015)

Could not the chance of the sex alleles be offset by rotating drones in from different yards or from different states from peoples hives that have shown a history of the mite tolerance that people are looking for and used in an AI program?


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## Fusion_power (Jan 14, 2005)

Stock transfer is a time tested and proven way to limit inbreeding effects. Read Randy's article and you will see that he is attempting to breed from his own bees as much as possible to avoid diluting the positive traits he has already concentrated. He will therefore limit the amount of outside stock brought in. This makes managing his gene pool much more important.


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## bucksbees (May 19, 2015)

squarepeg said:


> this is the latest in a series of articles some of which have been discussed here:
> 
> http://www.beesource.com/forums/showthread.php?332799-randy-oliver-article-in-january-17-abj
> 
> ...


Looks like it is now posted on his website, sweet.


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## Richard Cryberg (May 24, 2013)

Fusion_power said:


> Given the conditions, there are two correct answers. Under normal conditions, the daughter queen will have 1 allele in common with her mother and one from a drone. If the drone allele is different - which should be true most of the time - then the mating will result in 75% brood viability. If the drone allele is identical with one that the mother queen carries then the fact that a queen was produced automatically means the daughter queen has the same two sex alleles the mother queen carries. In this rare case, brood viability will be 50%. No matter the amount of inbreeding, it is not possible to get below 50% brood viability based on the sex allele.


Lets just say that the mother queen is Sd1^a and Sd1^b at the sex allele where the symbol Sd1 represents wild type. Yeah, I know Sd1 is not a valid symbol but I do not know the biological function of this gene and have not looked up its proper symbol. She is fertilized with the sperm of many drones. So, she could potentially carry sperm that are Sd1^ c thru however many sex alleles exist. As far as I know the number is not known. Older lit says about 20 and it seems pretty obvious this number is low. Randy tells me newer studies show a bunch more.

The net result is the daughter will be either Sd1^a and Sd1^c or Sd1^d, of Sd1^e .... or she will be Sd1^b and Sd1^c or Sd1^d, etc. That daugher is now fertilized by one a number of her brothers. Those brothers could be all Sd1^a or Sd1^b. Or they could be some mixture of those two sex alleles. So, in an very unlikely condition she could be fertilized with drones none of whom match her sex alleles and she would produce no diploid drones at all. Or she could be fertilized with all drones that match one of her sex alleles (also very unlikely) in which case she will lay 50% diploid drones. In the more probable cases she will be inseminated with a mix of drones and produce about 25% diploid drones. But, most any number between 50% and 0% diploid drones is possible. Of course this analysis ignores the possibility that the original queen was fertilized by some drones that were either Sd1^ a or Sd1^b. As she was laying a good pattern this possibility is easy to ignore and does not change the argument or numbers significantly. My point is proven. Randy understands the consequences of inbreeding and you have a bit to learn. In fact, inbreeding at the sex allele locus tells you very little about how inbred the queen really is overall, so just concentrating on the ill effects at the sex locus is a small enough part of the whole story as to be pretty meaningless in truth. You really need to consider the effect of crossing over, which Randy hints at in his article, to get a honest overall picture. An off the top estimate without doing the actual calculation is the above mating sequence would give a Wright's coefficient of roughly 30% which is fairly modest inbreeding. You are generally not badly inbred until the Wright coefficient gets up to 60 or 70% in most animals and in many species even that level is hardly consequential. If you are not inbreeding enough to see some queens laying an objectionable number of diploid drones you are not inbreeding enough to make any overall genetic headway towards other desired traits. The idea that seeing some queens laying too many diploid drones is a sign of trouble is simply not true. In a good breeding program it should be a hoped for result. Those just might be the best queens you raise in terms of value as breeders and it sure is not a reason to cull them.


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## Fusion_power (Jan 14, 2005)

Richard, read the literature and then you can make a reasoned argument. Under conditions where 20 drones from the mother queen are used to inseminate, the results are statistically in favor of the stated results where the mother and daughter share only one csd allele. But I see you missed the point of what I stated. The point was that when the mother queen and daughter have the exact same csd alleles, the percent diploid drone eggs will tend strongly toward 50%. This is still just a mental exercise with very little application in the real world. Randy will not be deliberately mating queens to their own brothers.

Documentation is available in the 2013 work of Lechner where the gene is referred to as "csd". The analysis estimated between 116 and 145 csd alleles worldwide in the western honeybee. Randy is - or will be - dealing with population genetics, not individual queens. The analysis he posted is based on segregation at the individual queen level. It is kind of like comparing a grain of sand to a sandy beach when comparing the genetics of one queen with the genetics of a population of queens. Don't get lost looking at trees, there is a forest out there.


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## JRG13 (May 11, 2012)

I dunno, all I know is I started a similar approach a few years ago and well, I should start telling it again in my thread when I have some time...


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## bucksbees (May 19, 2015)

If you go by the regional system that he is advocating, I am in the same as River, hopeful that would mean being able to trade queens to maximize saturation of mite resistance.


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## Riverderwent (May 23, 2013)

JRG13 said:


> I dunno, all I know is I started a similar approach a few years ago and well, I should start telling it again in my thread when I have some time...


You should. You spin a good tale.


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## Riverderwent (May 23, 2013)

Fusion_power said:


> IMO, progress can be made faster with a hard Bond approach.


Plus, it doesn't make your head explode.


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## Richard Cryberg (May 24, 2013)

Fusion_power said:


> Richard, read the literature and then you can make a reasoned argument. Under conditions where 20 drones from the mother queen are used to inseminate, the results are statistically in favor of the stated results where the mother and daughter share only one csd allele. But I see you missed the point of what I stated. The point was that when the mother queen and daughter have the exact same csd alleles, the percent diploid drone eggs will tend strongly toward 50%. This is still just a mental exercise with very little application in the real world. Randy will not be deliberately mating queens to their own brothers.
> 
> Documentation is available in the 2013 work of Lechner where the gene is referred to as "csd". The analysis estimated between 116 and 145 csd alleles worldwide in the western honeybee. Randy is - or will be - dealing with population genetics, not individual queens. The analysis he posted is based on segregation at the individual queen level. It is kind of like comparing a grain of sand to a sandy beach when comparing the genetics of one queen with the genetics of a population of queens. Don't get lost looking at trees, there is a forest out there.


I see the biochemistry is not understood and that is why we are stuck with the phenotype symbol csd. It is interesting that csd was picked instead of Csd. In general I would think Csd would be considered a better choice by most geneticists. The analysis did not show an estimated 116 to145 for western populations. That is an estimate for the world and speculative to boot being based on an extrapolation of existing data. It really makes little practical difference what the actual number is after you get past perhaps 30 or 40 unless you live on a small island that will only support a dozen colonies and never bring in any new blood for 50 years and there seems to be good data that says it is considerably larger than 30. Even on that small island it might not be that big a problem as some of the lit hints at pretty high mutation rates in the section of the gene responsible for sex determination. Very high mutation rates are well known in some situations. For a well understood human example consider Huntington's disease where the mutation probability is very close to 1.0 per generation per allele.

One problem in your analysis is your assumption that that the queen mother and her daughter are going to have the same two csd alleles. While possible, this is highly unlikely to happen. Remember, the daughter gets only one of her csd alleles from mom. The other came from some drone mom mated to. A daughter with the same allele compliment as mom would only happen if the queen happened to mate with one drone that had the same csd allele as she carried on one of her chromosomes. If the local population happened to have a total of 40 different csd alleles in the mix and all were in equal proportions and if she mated with 20 drones there is an even odds chance one of those drones carried the same csd allele as one of her copies and one daughter out of 79 from such a queen would have the same two csd alleles as the queen mother had. In this hypothetical there would be another queen that mated with zero drones where a common csd allele was shared so zero daughters like mom would be produced. Of course this hypothetical also allows one queen in 400 to mate with two drones which share one of her csd alleles (or is it one in 800? doing this in my head gets confusing) but by now the odds are so low I think it is fair to ignore those possibilities. The only other way a daughter can have the same two csd alleles as mom is if that daughter is the result of thelytoky, which is even more unlikely but not impossible either. In thelytoky the daughter is a clone of mom resulting from fusion of one polar body with the egg.

At any rate, while an interesting discussion, this has nothing at all to do with the consequences of what Randy proposes doing.

As an aside the honey bee should be unusually tolerant of inbreeding. Randy points out one reason in his article. Drones being haploid eliminate all recessive lethals and most sub lethals every generation so the species should be fairly clean compared to other species that lack such a filter. Much of inbreeding suppression in viability is the result of concentration of lethals and sub lethals. In many animals if you can get past the 7th to 10th generation of brother to sister mating vitality returns, often big time. There are other negative consequences of inbreeding in some species that prevent you from ever getting close to that far. Often those consequences result in sterile females but fertile males, both otherwise seeming to be quite viable and healthy. Both rabbits and pigeons are good examples where this happens. It appears in such animals some heterozygousity is needed for female reproductive success. In the pigeon case that heterozygousity must be autosomal, but in rabbits it could be on the sex chromosomes.


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## Fusion_power (Jan 14, 2005)

> As an aside the honey bee should be unusually tolerant of inbreeding.


 yes, but the devil is in the details. Evidence currently points to a high level of heterozygosity considered across the population of honeybees in a region. One estimate suggested it could be as high as 3% variability. The honeybee's method of mating retains this variability because the queen mates with about 17 drones on average. Breeding methods such as BLUP used in Europe have a distinct tendency to reduce population variance because they rely so heavily on single queen mothers to produce queens used to produce drones at mating stations. The brood viability rate based on the csd allele is used as a proxy to calculate inbreeding effects. I believe this is a problem, i.e. that over time they will find they have narrowed critical areas of the honeybee genome.

Randy's problem is different. He is relying on one single queen to establish mite resistance in his operation. Because this single queen's genome will become so prevalent, he will wind up with a much reduced effective population size and a significant increase in deleterious effects caused by the csd allele. The net effect would be like taking 2500 colonies and halving them. The effective breeding population is only 1250 yet there are 2500 colonies to be managed. What he needs badly at this time is an unrelated highly mite resistant queen to reduce the effects of using a single queen as the foundation of his breeding program.


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## Richard Cryberg (May 24, 2013)

Fusion_power said:


> Randy's problem is different. He is relying on one single queen to establish mite resistance in his operation. Because this single queen's genome will become so prevalent, he will wind up with a much reduced effective population size and a significant increase in deleterious effects caused by the csd allele.


Sorry, but loss of adequate csd variance is simply not going to happen for Randy. Forget this idea. It is just plain wrong for all kinds of reasons. If anything he will have the opposite problem of too much variance showing up. In fact I think that is why his program is likely to fall far short of producing reliably bullet proof queens with respect to mites. He will produce better queens but not bullet proof. He will do far better than any back yard effort, but still not bullet proof.


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## lharder (Mar 21, 2015)

Probably bullet proof should be abandoned as a concept. No such thing. Even things like tracheal mites and AFB cause problems from time to time. Again hard bond selection keeps things in check. The goal is more reasonable mite/virus control with hard bond keeping things in check. If you don't have some losses, then not enough selection is happening and weak bees are being kept alive. The question is what is acceptable loss. My close to 50% is too high long term but is somewhat sustainable based on my increased hive numbers. However, with time I would like to see that go to 20 to 30 %. I could see that happening as my foot print gets bigger, I get more control over the genetic space, and I go through more iterations of selection. Too much chaos is my enemy though, so it could all fall apart yet.


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## Richard Cryberg (May 24, 2013)

I should probably expand on my statement that a female bee produced by thelytoky is a clone. A frequent definition of clone is an individual produced asexually from one individual and genetically identical to that parent. This definition is always wrong. A clone will probably never be genetically identical to the parent. Even a clone formed by grafting a plant or rooting a plant cutting is not genetically identical to the parent stock if you look at the full DNA compliment and particularly not identical if you look at epigenetic marks. There are many reasons this is true that are well understood, and probably many reasons we do not even know about yet. A more accurate definition in view of what is known today about genetics would be any individual produced asexually from a single parent.

With that in mind during thelytoky crossovers have happened before the cells divide and form polar bodies. So, all kinds of DNA swaps between chromosomes have happened. When a cell that should have been a polar body fuses to the egg and forms a diploid cell you have the equivalent to a fertilized egg which can develop into a normal individual. Half of all such events will result in a fertilized cell that contains identical csd alleles and will be a diploid drone. The other half will have the same two csd alleles that mom had and will develop into a normal female.

For those whose heads blow up over trying to understand this stuff I have sympathy. I will tell a short story. One of my best friends is a DNA jockey college prof. When he teaches the first graduate level course in genetics to students the first day in class he tells them "You think you know something about genetics. By the end of my third lecture you will realize you know nothing about genetics." He tells me he is seldom wrong. Fortunately political correctness has not yet totally taken over teaching at the grad level or he would get in trouble for saying this to students. I could teach parts of that course better than he can due to my stronger back ground in things like physical chem, org chem and quantum mechanics. Ideas like "consider the implication of the keto to enol equilibrium of adenine and how that impacts T -> C errors during replication" are intuitively obvious to me but total Greek to most BS biologists. Yet, those kinds of thoughts are integral to any serious thinking about modern genetics. In good modern genetics courses, even at the undergrad level all that tedious Mendelian stuff is barely mentioned. It is worth about two pages in the text book. Not long ago it was the whole course and with the load of details that required rote memorization the normal student promptly forgot much of what was taught when the course was done. When you learn the subject from a chemical standpoint the Mendelian stuff is an automatic consequence and does not need any teaching time as there are much more important concepts you need time to learn. The downside is that for the general public their heads explode. Science has become terribly complex and there is no way to make that complexity simple. This is a major problem for scientists too. I read a few hundred pages of science stuff a week and am terribly dated in most areas.


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## squarepeg (Jul 9, 2010)

so i've finally got most of my hives settled in and in good shape strength wise and equipment wise to exploit the remainder of our main spring nectar flow. i'm now going back for a more careful read of randy's latest installment in his 'the varroa problem series'.

the first thing i came across that i feel is noteworthy and appears to hit home with respect to what a number of us are reporting here on the forum is this:

"In my case, I decided to return to my old ways, and to work with what I already had — locally-adapted stock selected for characteristics that I found desirable. To that, I brought in the occasional instrumentally-inseminated VSH, Russian, and a few other promising breeder queens to produce drone mother colonies.

Practical application: by adding some of those drone mothers to my mating yards, i’d “offer” the new alleles (and genetic and epigenetic combinations) to my own breeding population without necessarily messing up the fine tuning that population already had going. If the new combinations proved to be adaptive, they’d make it into future generations; if not, they’d be bred out, since I wouldn’t select queens from second-rate colonies as breeders."

abj may 2017 vol.157 no. 5 p. 510

randy appears to be upholding the validity of utilizing stock that is already adapted to and showing success in local area as a good starting point.

there are several here lucky enough to have a thriving feral population in place and have been able to capture and propagate from them. even luckier, we are finding that they are productive and easy to work with.

while i am fairly sure that my reared queens are mating with drones from the local ferals, i am finding that a few years of selecting/deselecting is enough to make gains in survival, productivity, less swarminess, ect., and apparently without sacrificing anything in the mite resistance department.

this is all well and good, and seems like a no brainer for those in ecoregions supporting a thriving feral population, but what to do for those in areas in which there doesn't exist a resistant feral population?

back to the article.


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## 1102009 (Jul 31, 2015)

squarepeg said:


> this is all well and good, and seems like a no brainer for those in ecoregions supporting a thriving feral population, but what to do for those in areas in which there doesn't exist a resistant feral population?


Why, we can do just like you SP, you never know if the ferals ( if they are) really are resistant or if it is your breeding and beekeeping management!
Maybe we have more losses, maybe we need more time, but losses are normal and time will tell. 

Even a small beekeeper can do selection and as a group we can accept higher losses.


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## lharder (Mar 21, 2015)

squarepeg said:


> so i've finally got most of my hives settled in and in good shape strength wise and equipment wise to exploit the remainder of our main spring nectar flow. i'm now going back for a more careful read of randy's latest installment in his 'the varroa problem series'.
> 
> the first thing i came across that i feel is noteworthy and appears to hit home with respect to what a number of us are reporting here on the forum is this:
> 
> ...


Yes that is the same conclusion I have come to. Test local populations to see what tools the population has to work with. Only bring in bees that have traits that are missing from the local population. I would be a bit more patient that Randy in that I would like to see that trait expressed at some point in my local bees at some frequency. Once established, survival and production would determine the frequency of expression in the local population and would tell you how useful that trait is. But it would come at a cost of introducing some chaos into the local system for a time. I can imagine one set of bees occupying one solution space, and another occupying another solution space, but a mixture of the two being in no man's land. But at the same time there are a few genetic quirks that allows some mixing and survival. But much death is likely.

So I am not sure that there are many feral bees in this neck of the woods. But I think there are some decent genetics lurking out there as I have found some that do ok. The question is what frequency is it expressed in the local population? So as I make daughters from survivors, what percentage of them will turn out to be ok after 2 years or more? My long term success is likely determined by this. Also the data is very messy. There are some site considerations and beekeeper inexperience. My methods are evolving or devolving depending on the viewpoint. 

This all said, I would hazard that my sites that has selection pressure, would find the bees that occupy a solution space and increase the frequency at a site. As long as these sites are stocked with bees that experience selection pressure, they would have influence on nearby populations of bees that are allowed to raise queens, treated or not. So as the number of my sites grow and the nodes of selection increase, and eventually I would hope to find increased survival.


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## mike bispham (May 23, 2009)

Fusion_power said:


> Randy notes that he is trying to avoid doing a hard Bond test which has serious financial side effects.


Do we know why Randy doesn't try John Kefuss' recommended 'soft bond' approach for commercial beekeepers, using frozen brood (and other) assays?

Mike (UK)


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## mike bispham (May 23, 2009)

Riverderwent said:


> Plus, it doesn't make your head explode.


 As John Kefuss says, you don't need to know _why_ some things work, you just need to know _that_ they work.

Mike


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## AstroBee (Jan 3, 2003)

squarepeg said:


> "In my case, I decided to return to my old ways, and to work with what I already had — locally-adapted stock selected for characteristics that I found desirable. To that, I brought in the occasional instrumentally-inseminated VSH, Russian, and a few other promising breeder queens to produce drone mother colonies.
> 
> 
> randy appears to be upholding the validity of utilizing stock that is already adapted to and showing success in local area as a good starting point.


SP,

My subscription to ABJ has expired, so I can't yet dig into the details of this article. However, looking at your quoted text above, I see that his approach is broader than working with "locally-adapted stock" as he followed that statement with referencing his targeted importations. Again, I'm only reading the quote above, but it seems to me broader than "locally-adapted stock". I believe that relying only on locally-adapted stock (with potentially unknown resistance) is going to be a tough (and possibly long) road for some. My path forward heavily leveraged importing very good quality stock and using it with the best of my survivor stock.


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## squarepeg (Jul 9, 2010)

i may be oversimplifying what was written in the article but...

i believe randy is monitoring mite counts on hundreds of hives and moving the ones with low counts and zero counts to special isolated 'breeding' yards that are kept off treatments. 

from this stock he chooses his breeder queens and in these yards he sets up his mating nucs to get the drone contribution.


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