AMM 99% pure against what?

[TooBee...]

Hearing of the recent BIBBA conference reminded me of this nagging question I've had for some time.

I was at a lecture in which the speaker mentioned that his AMM's ("Native" Dark Bees) DNA were 99.2 to 99.7% pure (in the strict Native sense, not the general AMM sense, if you see what he was meaning...), I didn't want to ask how he knew this as I didn't want to show my ignorance. He had got them DNA tested, that's fine, but my confusion is; to know that they are 99% pure doesn't he need to know, 'pure against what'? I mean if you are going to test the DNA and declare that it is 99% the same as a pure "Native" AMM, then surely you must have a sample of a 100% "Native" AMM to test it against, and since we have been importing European bees (including some dark African mountain bees) for many centuries, especially on mass since the 1920's, well before DNA testing, how can one say that their bees are 99% identical to a bee that could never have been tested against, and cannot for certain be found nor tested against?

My confusion again was sparked some months back after reading this paragraph on page 14 in the following PDF.
"Recent surveys report honey bees of native type in 14 counties of England and Wales and all but two of the Scottish Regions (Pritchard, 2006; Stoakley and Stoakley, unpublished). Samples from at least six of these have had their identity as
A.m.m. confirmed by DNA analysis".
https://bibba.com/wp-content/upload...-Bee-really-native-to-Britain-and-Ireland.pdf
(Bold emphasis added by me for clarity)

The fist sentence is based on unpublished research??? The second sentence would be easy to give a reference for and publish it?

I'm just a bit confused, and I know that there are some here that are much more familair with this kind of stuff than I.

Please do not allow this Post start an argument, please make it an opportunity for some of us to increase our knowledge, or at least to learn of other beekeepers' points of view, even if some of us may disagree with them

PS: You will note that I have placed this in the Beginners Section as I suspect this is a basic bee DNA question that I have asked, which will show my beginner ignorance on the subject.

 

[deemann1]

Let me guess, you don't keep AMM

 

[SDM]

I suspect the best that they can do is confirm an absence of markers from known European Amm populations. Though I suppose it's possible that a DNA sample could be taken from a museum display that predates imports to use as a baseline.

 

[masterBK]

Didn't someone do some DNA tests on remains of bees from Viking times found in York?

 

[TooBee...]

Let me guess, you don't keep AMM

Guess again....

at least I think I do, they're pretty dark.

 

[TooBee...]

...Though I suppose it's possible that a DNA sample could be taken from a museum display that predates imports to use as a baseline.

Is that possible, I didn't think it was. Do any of you have links to that?

Also I'm still not 100% convinced that would do it, I mean, the 'sample' in the museum could have been brought over by Vikings or Monks, but then maybe by 'Native' it's meant that their ancestry line has been here for several hundred years. Or maybe it's that the British / Irish AMM strain / descendants have DNA markers that are no longer present in their continental brothers (died out on the continent but survived here). But I'm guessing, without someone else with more knowledge on this joins in.

 

[For want of a better word]

From my own experience ...
There seems to be a variance in the "look" ( Morphological appearances) within the Amm in the UK... Manx Amm look different to ones from Northumberland and Amm from Cornwall.

My understanding on the purity percentages given by the laboratories ( eg Apigenix in Switzerland) is that the genetic markers for non Amm sub species found within the honeybee DNA that is being sequenced give the percentage "purity" and hence the level of introgression that has taken place.

A lot of misunderstanding concerning Nuclear DNA and mitochondrial DNA seems to have been used by some to muddy the waters on "purity "levels.

Mitochondrial DNA is conserved through the female line within mitochondria in EGGS... there is no mitochondrial DNA transfer from sperm***

Mitochondrial DNA can be used to look into the bees parentage to see the level of introgression from the past generations ( none...usually denoted as M group (Apis mellifera mellifera) or some C group A m ligustica / carnica [Italian/ Carnolian)

I am not convinced that a comparison with a museum specimen is of much help given that DNA nucleotide constantly change over time to some degree.

*** there are mitochondria in sperm as they are the drivers that administrate energy to the flagellum... in some extremely rare cases mitochondrial DNA has been found to have been inherited in humans.... due to the mitochondrial DNA from the sperms tail getting into the egg... it happens!

Once again from my own experience the purity level in any sub species does not guarantee levels of honey production of lack of aggresiveness or swarminess... I have seen "pure" A m c that have been bitches from hell ' and jet a hybrid between it and a A m l have been productive little pussey cats!!

Many will disagree as they see DNA analyses as a "faith system" rather than hard fact Science!!... and will jump through hoops of fire to disagree with me with accusations of trolling etc etc

Yeghes da

 

[B+.]

Many will disagree as they see DNA analyses as a "faith system" rather than hard fact Science!!... and will jump through hoops of fire to disagree with me with accusations of trolling etc etc

I'm curious what you are performing the analysis on.
If you test drone eggs, you test 16 of the queens 32 chromosomes. This is the approach taken by Dr. Anja Strauß and Prof. Dr. Kaspar Bienefeld on the GeSeBi project.
If you test workers (or queens), you're testing 16 from the queen and 16 from the drone that fertilized the egg.
Which do you test?

 

[JEP]

The concept of a sub-species isn't really something that can be quantified by a percentage DNA analysis even though some studies try to do just this.

When Carl Linnaeus classified Amm in 1758 he had no concept of DNA, or the DNA purity of a sub-species. He just noted some morphological characteristics of honey bees around him in northern Europe (Germany perhaps?) and gave them a name. He didn't give bees in general too much thought as he was too busy classifying anything else he could get his hands on.

So a subspecies is more an opinion rather than something that is set in stone. While there are clear obvious morphological differences between bees from different areas e.g bees from Italy tend to be more yellow in color etc. these are still only someones opinion of what should form a subspecies. At the end of the day all the bees we as beekeepers are concerned with are the same species. This means that even the DNA from different bee sub-species is going to be at least around 99% identical. If you doubt this then consider that your own DNA is 96% identical with that of a chimp - an animal from a different species.

So determination of DNA purity in the sense of your original question means test a load of bees that look like Amm is supposed to look to determine what a pure sample is and find common DNA markers. Then use these markers to see if further bees you test fit this profile. As you imply this then can only give you a purity related to those bees originally tested.

As bee sub-species were only ever based on visual assessment then by the strictest definition if a bee looks like an Amm type bee and ticks all the appropriate morphological requirements then it is Amm.

 

[For want of a better word]

I'm curious what you are performing the analysis on.
If you test drone eggs, you test 16 of the queens 32 chromosomes. This is the approach taken by Dr. Anja Strauß and Prof. Dr. Kaspar Bienefeld on the GeSeBi project.
If you test workers (or queens), you're testing 16 from the queen and 16 from the drone that fertilized the egg.
Which do you test?

Tests have been carried out using antennae from 16 drone ( brother) larvae, and on queen wing clippings and on single (worker) samples.... so both full genomic sets have been analysed.
I can only refer you to the B4 webpages for more info.
Can PM you a contact number for someone who has a lot more and better information than I have!

Purity it must be said is academic!

Yeghes da

 

[TooBee...]

B+
it was my understanding that it was the 'bees' being tested or it could be construed, within the context of the lecture that it was the Queen bees specifically. I noticed a slight change in tone and hesitancy in the speakers voice when he stated his 99+% figure, I have often observed this can be a sign of a lack of certainty (or to be less kind, a knowledge that what is being said isn't 100% accurate).

JEP
that's an excellent point! I had heard that a male chimpanzee was 99% (you say 96%) identical to a male human, therefore one should not be surprised to find near 99% identical DNA from a pure 'native' AMM to that of any local wild dark looking bee.

 

[B+.]

Tests have been carried out using antennae from 16 drone ( brother) larvae, and on queen wing clippings and on single (worker) samples.... so both full genomic sets have been analysed.
I can only refer you to the B4 webpages for more info.
Can PM you a contact number for someone who has a lot more and better information than I have!

Purity it must be said is academic!

Yeghes da

This is a little confusing
1. If the drones were brothers, rather than sons, you are testing the DNA of the queens mother rather than the queen herself (i.e. the queen that laid the egg). If the queens mother wasn't pure mated, the queen (and her progeny) wouldn't be either.
2. Samples from female progeny are dependent on pure mating (for the reason explained above). This kinda depends on using II in this country.
3. The wing clipping tests the DNA of the queen but care would need to be taken to ensure she was control mated, otherwise, you could only be sure of the male progeny.

What I'm trying to say is that it's all well and good testing the DNA of your stock, but, you also need a controlled breeding programme or the benefit of testing will be lost. You also need a way of identifying individual queens so the ancestry can be plotted. Do you have all this?

 

[B+.]

B+
it was my understanding that it was the 'bees' being tested or it could be construed, within the context of the lecture that it was the Queen bees specifically. I noticed a slight change in tone and hesitancy in the speakers voice when he stated his 99+% figure, I have often observed this can be a sign of a lack of certainty (or to be less kind, a knowledge that what is being said isn't 100% accurate).
.

Thank you. I think Icanhopit/Cheers clarified that point by saying it was clippings from the queens wings. I suspect that this means they were instrumentally inseminated first although a naturally mated queen could also have its wings clipped.
I can't comment about the credibility of the speaker because I wasn't there.

 

[TooBee...]

Hi B+

the 'native' AMM Queens being referred to as being 99+% pure were all Open Mated, it was most definitely not an Isolated Mating Station, I think you're absolutely right about what you say (and have said in other Posts) about II, I'm not aware of anyone here in Ireland that is doing II and providing their Queens for sale (although I think there is a group far south that may be starting / doing it amongst themselves).

The speaker (which I don't want to mention) is well known here (especially in 'native' bee circles) and I have only heard excellent things about his character - so I don't believe there was any intention to mislead: In fact I would go a little bit further, in that I don't believe the Very Vast Majority of those (here in Ireland at least) that promote the local 'native' AMM are trying to mislead, the more I learn about all this, I think they are just mis or ill informed.

I would love to get into II, but that's definitely a few years away!

 

[jenkinsbrynmair]

So basically, you are not really trying to get the facts, but using the beginner's section as some kind of protection as you attempt to do a bit of nifty trolling of the AMM enthusiasts.

 

[For want of a better word]

This is a little confusing
1. If the drones were brothers, rather than sons, you are testing the DNA of the queens mother rather than the queen herself (i.e. the queen that laid the egg). If the queens mother wasn't pure mated, the queen (and her progeny) wouldn't be either.
2. Samples from female progeny are dependent on pure mating (for the reason explained above). This kinda depends on using II in this country.
3. The wing clipping tests the DNA of the queen but care would need to be taken to ensure she was control mated, otherwise, you could only be sure of the male progeny.

What I'm trying to say is that it's all well and good testing the DNA of your stock, but, you also need a controlled breeding programme or the benefit of testing will be lost. You also need a way of identifying individual queens so the ancestry can be plotted. Do you have all this?

The drone antennae ( in fact from 30 brood drones from same brood comb) tested were laid by a queen that had been DNA analysed as >98% Amm on the nuclear DNA and was Amm in the mitochondrial DNA.... as was her mother.
The queen's( mothers) DNA was reflected in them as being the same in both Nuclear and mitrochondrial.
The mating in this particular instance was on a drone flooded penninsular , so as you say without II we have no certainty!

This season we have had the fortune to be given access to a very isolated site... on fact when virgin queens were released there, none got mated.

Nest season we now have 3 breeders to get going with II....

My own hybrid of Northumberland drone sperm suppliers with Manx virgins (II) look interesting... still in nucs, but seem to be calm... the purists will be having an fit!!!....
not in my "isolated valley"...
which does not look so isolated as a "new" beekeeper brought in stock that I did not know about... only one small colony presumably to pollinate his pollytunnell and allotment.... against 50 full blown Amm colonies all pushing out drones.

 

[B+.]

So basically, you are not really trying to get the facts, but using the beginner's section as some kind of protection as you attempt to do a bit of nifty trolling of the AMM enthusiasts.

Not necessarily JBM.
I've had some private correspondence with TooBee and he seems genuine in his desire to learn. However, learning involves a lot of questioning so don't be so defensive. I asked lots of questions when I was in Bibba too. This ultimately led to me leaving, but, that's not every ones choice

 

[TooBee...]

So basically, you are not really trying to get the facts, but using the beginner's section as some kind of protection as you attempt to do a bit of nifty trolling of the AMM enthusiasts.

That's not fair!

Before you point a finger, look and see the three pointing right back at you.

Through Googling the terms being used by the positively contributing members on this thread I have learned quite a bit of new information. II is one major part of beekeeping which really fascinates me, and I want to take these couple of years before I start it to learn as much as I can before I start being taught it, if you see what I mean.

PS: Changed your Avatar? Let me guess, you're the one surrounded by the men in the white suits!
oh, I'm so sad, I'm chuckling at my own joke!

 

[Quis Custodiet]

That's not fair!

Before you point a finger, look and see the three pointing right back at you.

Through Googling the terms being used by the positively contributing members on this thread I have learned quite a bit of new information.......................

Don't let Jenkins upset you.....that is what he wants to do. Remember he is more deserving of pity than contempt.....poor fellow thinks he is Nelson re incarnated.

 

[jenkinsbrynmair]

learning involves a lot of questioning so don't be so defensive.

It doesn't involve the snidey little throwaway remarks at the end though.
Perrsonally, I have no reason to get defensive, just making observations from the sidelines.