# FREE tools for honey bee wing morphological analysis.



## GregB (Dec 26, 2017)

There is another Russian-only tool - "Breed by the Wings" - was developed a guy interested in pure AMM selection and breeding.
This tool was actually the prototype for the MorphoXL above - they have some differences in the analysis outcomes (not for me to judge which is more correct).





«Порода по крыльям» | Жил-был пчеловод


ИНСТРУКТИВНО-МЕТОДИЧЕСКИЕ МАТЕРИАЛЫ по работе с программой определения породной принадлежности семьи пчел «Порода по крыльям» 1. Общие положения 1.1. Настоящая программа разработана в среде Excel-2003, нормально функционирует в EXCEL следующих выпусков, вплоть до входящего в состав Office-2010...



www.lifeandbees.ru




I am slowly working on making "Breed by the Wings" English-user usable eventually.
I like this tool better than then MorphXL and prefer it (but use both for cross-checking the analysis results).


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## Litsinger (Jun 14, 2018)

GregV:

I appreciate your work with this, and for your willingness to take the time to post it here for our use and discussion.

Russ


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## GregB (Dec 26, 2017)

Found this interesting article:








Wing Morphology - How Native are my Bees?


In the spring of last year I decided to investigate how native my bees were, so I conducted wing morphology on all my colonies, a technique which involves measuring positions of vein intersections on the bee’s main wing. The results were a surprise, as it looked like my bees were native, or at...



www.northumbrianbees.co.uk





It points to another interesting product - CBeeWing (trial is free).


Using CBeeWing for better bees


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## joebeewhisperer (May 13, 2020)

Greg,

You had described such a program a few months ago (scan a wing, map the points) and somehow I landed on IdentiFly | DrawWing , the software being IdentiFly. This is a zip file. You unzip and run the executable. It's pretty sophisticated. I got the developer's info and sent him a bit of cash as this took some work. However, there was nothing anywhere asking for money. I think maybe this guy is a researcher/student in Poland. 

I actually thought it was the one you were referring to then.


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## GregB (Dec 26, 2017)

joebeewhisperer said:


> I actually thought it was the one you were referring to then.


I looked at IdentiFly and passed (and mentioned about it too).
Too complicated; too much work.
Even with 8 point (per a wing) analysis it takes much time already.
18 point analysis - no thanks.

Besides, what extra information the 18 point analysis will give me?
The the main three indexes of the 8 point analysis output are plenty information to chew on.


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## GregB (Dec 26, 2017)

I'd be interested in samples of pure Ligustica and/or Carnica if someone has any.
I am talking of pure enough grade - good enough to calibrate/verify the tools.
Not just kinda/sorta Italians/Carni - those are most likely mutts (useless for calibration).


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## HaplozygousNut (Dec 30, 2015)

GregV said:


> I'd be interested in samples of pure Ligustica and/or Carnica if someone has any.
> I am talking of pure enough grade - good enough to calibrate/verify the tools.
> Not just kinda/sorta Italians/Carni - those are most likely mutts (useless for calibration).


GregV, do you think you could also get some wings of some A. m. mellifera from different regions, like the Spanish and Italian strains of German black bees? I was wondering whether they'd be slightly different in the wing veining from the Russian strain of Amm...

-Nathaniel Long


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## GregB (Dec 26, 2017)

HaplozygousNut said:


> GregV, do you think you could also get some wings of some A. m. mellifera from different regions, like the Spanish and Italian strains of German black bees? I was wondering whether they'd be slightly different in the wing veining from the Russian strain of Amm...
> 
> -Nathaniel Long


I don't know anyone in Spain/Italy to ask for those particular wings (not to mention the wings should be trustworthy as true AMM wings).
In Russia I hardly know anyone either, but at least they post their wing data online (which include the index numbers that I could use).

The 8 point wing analysis is not precise enough to be sorting out AMMs from AMMs - that is not a right tool for the job.
It is only good enough to separate AMMs from the others as far as I know, like so:


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## HaplozygousNut (Dec 30, 2015)

GregV, I found a photo (Європейська темна бджола — Вікіпедія) on the Ukrainian Wikipedia German black bee article that has wing veining shape that looks like the wing image on this website (A Comprehensive Characterization of the Honeybees in Siberia (Russia) | IntechOpen Figure 2) of "zero" discoidal shift, different from the wing image of "negative" discoidal shift. Sorry if it is hard to understand what I am saying, I don't know the terminology for wing veining.


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## GregB (Dec 26, 2017)

HaplozygousNut said:


> GregV, I found a photo (Європейська темна бджола — Вікіпедія) on the Ukrainian Wikipedia German black bee article that *has wing veining shape that looks like the wing image on this website (A Comprehensive Characterization of the Honeybees in Siberia (Russia) | IntechOpen Figure 2) of "zero" discoidal shift, different from the wing image of "negative" discoidal shift. *Sorry if it is hard to understand what I am saying, I don't know the terminology for wing veining.


I know exactly what you mean.
The wing on your picture is not looking like a commonly known AMM wing (negative DsA). Looks more like a Carni/Ligustica wing, just by a general look of it (zero to positive DsA).

However, a single index is not sufficient to make a call.
Here attaching a visual of Cubital indexes from my mutts (the extreme cases from one colony sample) - a couple of very much Millifera-like and a couple of very much Carnica-like.


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## Litsinger (Jun 14, 2018)

GregV:

While all this stuff is above my pay grade, I do enjoy and appreciate you posting about your studies. 

It is an interesting evaluation tool, and I will look forward to it hopefully yielding something of actionable benefit for you.

Keep up the good work-

Russ


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## GregB (Dec 26, 2017)

Litsinger said:


> GregV:
> 
> While all this stuff is above my pay grade, I do enjoy and appreciate you posting about your studies.
> 
> ...


The more I do these the more I realize - the morpho-tools (and the morpho-analysis as a whole) are great when trying to quickly isolate pure-enough breeds.
That was the whole original intent of this type of work.

Well, I am yet to see pure-enough breeds (granted I only the beginner, but the trends are becoming visible).
The real issue for the North American case - we deal with random hybrids left and right.
With few exceptions, I suspect there is no pure bee around here to speak of (outside of the marketing buzz, mis-information, and outright lies).

For example, there is no very obvious difference between almond bees and VSH (TM) bees.
They are roughly very similar hybrids of Carnica/Ligustica/Caucasica.
Maybe some clean sampling and tight statistical analysis can produce some significant differences which takes some sophistication.

Trying to wrap my head around looking at the data I get.
There is a need for totally different approach - identification and classification of certain hybrids (vs. the pure bees).


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## Litsinger (Jun 14, 2018)

GregV said:


> With few exceptions, I suspect there is no pure bee around here to speak of (outside of the marketing buzz, mis-information, and outright lies).


GregV:

It would be interesting to see how the morphological analyses might compare if you looked at representative stock from various regions of the country, particularly from more rural and less horticulturally-intensive areas to see if these populations demonstrate significant variation versus what you suspect locally are cast-offs from migratory beekeeping operations.


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## GregB (Dec 26, 2017)

Litsinger said:


> representative stock from various regions of the country,


What is exactly the "representative stock", Russ?
Such thing exists?
And who are those people who maintain this stock (not a small claim if to be done)?


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## Litsinger (Jun 14, 2018)

GregV said:


> What is exactly the "representative stock...


GregV:

I don't mean over much by this- strictly wondering- 'is the prevalent bee stock in my area (as an example) morphologically similar to your stock (as an example). If so, it supports the idea of a fairly monolithic admixture of genetics in the US. 

If not, it might suggest regional genetic variation supporting a survival advantage in a specific setting, particularly when not overwhelmed by annual genetic imports.


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## GregB (Dec 26, 2017)

Litsinger said:


> GregV:
> 
> I don't mean over much by this- strictly wondering- 'is the prevalent bee stock in my area (as an example) morphologically similar to your stock (as an example). If so, it supports the idea of a fairly monolithic admixture of genetics in the US.
> 
> If not, it might suggest regional genetic variation supporting a survival advantage in a specific setting, particularly when not overwhelmed by annual genetic imports.


Btw, if you have any per-annual surviving stock this spring, maybe we should measure them?
It should be 3-4 years you've been running them off chems, yes/no?
Why guess if we can just look at them.


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## Litsinger (Jun 14, 2018)

GregV said:


> ... maybe we should measure them?


It's a bet, GregV. Even better, why don't you plan a trip to Kentucky Lake for the first week in May and I'll send you home with some Kentucky feral stock to see how they fare in your harsh Wisconsin environment?


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## GregB (Dec 26, 2017)

Litsinger said:


> It's a bet, GregV. Even better, why don't you plan a trip to Kentucky Lake for the first week in May and I'll send you home with some Kentucky feral stock to see how they fare in your harsh Wisconsin environment?


Well, that is some trip from here. Dunno.
But maybe we can talk a queen or two via USPS, eh?
We should talk.


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## Litsinger (Jun 14, 2018)

GregV said:


> ...maybe we can talk a queen or two via USPS


If I would learn how to get queens without swarms I'd be glad to do it. I must admit that I am a PPBK.


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## HaplozygousNut (Dec 30, 2015)

GregV said:


> I don't know anyone in Spain/Italy to ask for those particular wings (not to mention the wings should be trustworthy as true AMM wings).
> In Russia I hardly know anyone either, but at least they post their wing data online (which include the index numbers that I could use).
> 
> The 8 point wing analysis is not precise enough to be sorting out AMMs from AMMs - that is not a right tool for the job.
> ...


GregV, do you think it would be a good idea to measure the tongue lengths of your bees that come up as high percentage Caucasica on the wing-morphometrixs? Caucasian bees are unusual in having long tongues. I have wondered about whether the bees coming up "Caucasica" in the wing morphometric tool are actually something else, like an Amm hybrid... I know that Amm genetics (or "M" lineage genetics) have been found here in the Southeast US, and I have heard about Thomas Seeley's feral bees in Ithaca, NY having a good percentage of Amm. So, it is hard to think Amm. would be so uncommon as is your experience with testing the wings of your local bees in Wisconsin.

Also, do you think hybridization could cause problems with the "negative" discoidal shift wing vein shape? Perhaps if the "negative" discoidal shift is a recessive trait, then it would be easily masked and not show up, even when close to pure Amm?

-Nathaniel Long


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## Struttinbuck (Mar 8, 2020)

GregV said:


> I'd be interested in samples of pure Ligustica and/or Carnica if someone has any.
> I am talking of pure enough grade - good enough to calibrate/verify the tools.
> Not just kinda/sorta Italians/Carni - those are most likely mutts (useless for calibration).


are you east coast or west? Sue Colby West for Carnica And Jason Bragg East.


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## GregB (Dec 26, 2017)

HaplozygousNut said:


> GregV, do you think it would be a good idea to measure the tongue lengths of your bees that come up as high percentage Caucasica on the wing-morphometrixs? ...........
> 
> Also, do you think hybridization could cause problems with the "negative" discoidal shift wing vein shape? Perhaps if the "negative" discoidal shift is a recessive trait, then it would be easily masked and not show up, even when close to pure Amm?
> 
> -Nathaniel Long


Every single additional measurement is a hassle in itself.
I don't care to chase after every rabbit.
Rather would just become good at something useful and give it a try.

In theory, sure - every additional measurement is a data point IF done correctly and conforms to some known standards (if done incorrectly it is just noise and time wasted).

I am not ready to comment on the wings yet (DsA trends, etc)
Have two last datasets to measure for myself and then sit back and look at them.
And read some resources.

Indeed, Caucasica and Millefera are kind of close to each other vs. the Ligustica/Carnica - per the wing morphometry.
So, granted we here largely work with the hybrids (not pure bees), thinking I will be just using the most distinct reference groups: Mellifera, Caucasica, Ligustica, Carnica (as depicted).
Any more additional references/finer resolutions don't really add more clarity to the picture.


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## GregB (Dec 26, 2017)

Here is a sample that is suggestive of Caucasian mix by its Cubital Index distribution.
But the Hentel Index/Discoidal Anglel pull towards Ligustica/Carni.
So this one could be, indeed, a mix of Caucasian/Carni
These bees are of dark disposition too - suggestive of Caucasian/Carni mix.


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## HaplozygousNut (Dec 30, 2015)

Okay, nice! Thank you for all the good information, GregV!


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## GregB (Dec 26, 2017)

Struttinbuck said:


> are you east coast or west? Sue Colby West for Carnica And Jason Bragg East.


Frozen Midwest.


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## msl (Sep 6, 2016)




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## GregB (Dec 26, 2017)

Like I said, MSL, 18-point wing landmarking is a very skill-dimending and slow project (what they refer to in the video).
Unless you are in academia, I don't see folks bother with it.
Good land-marking is a true skill and a challenge.

A couple nights ago I tried to landmark a wing set - wow - I could hardly do it because I was tired and it was late (and maybe just botched the data set anyway). Talking the simple 8-point landmarking, of course.

But this 18-point complication is really not needed IF you are simply trying to average out your indexes between few very main lineage groups - Mellifera (M), Caucasica (O), and Ligustica/Carnica (C) to see where your bee maybe landing.
I don't particularly look for Cyprian or Anatolian bees in my wings. 
Looking for traces of Scutellata sounds interesting, but I'd leave it to some enthusiast.

Really, I am rather interested in documenting the main index (CI, DsA, HI) distributions for the bees that seem to do well - does not matter what they are. No particular need in identification of the possible underlying sub-species under the conditions of massive hybridization (though it is interesting and how the tools are conventionally setup - to try to find the underlying sub-species).

The real value (in my view) is about separating some particular hybrids from other particular hybrids based on some factual numerical data (in addition to the human observation data).


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## Gray Goose (Sep 4, 2018)

Very interesting work Greg, And much appreciated.

I do wonder IF the VSH "trait" can be a learned trait or part of some brain feature. more memory for example.
Also M Bush mentions up to 800 different things living in a bee hive so some of these would be good and help with immunity and some like the Virus are bad. 

I do believe the genetics can be derived from the wing analysis, however that is only 1 of perhaps several involved in bee health. And Mite resistance.

Even Apiary location seems relevant, hence moving a "TF capable bee" and it does not perform the same.

As always more data leads to more questions.

Keep up the approach, an expert in this area is needed IMO

GG


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## GregB (Dec 26, 2017)

Gray Goose said:


> Very interesting work Greg, And much appreciated.
> ..........
> GG


Thanks GG.
There is tons to learn.
Yes, there are tons of variables.
Unfortunately, most of these go at the level of anecdotes (vs. hard factual data).
For example, I'd love to know few hard facts about M. Bush's bees as well as the surrounding situation - but don't hold much hope to ever hear of those.


I got entire online community of morpometry folks (some are true experts) I am trying to follow.

Here are Russian enthusiasts about the true AMM and they do a lot of wing analysis.
The genetic analysis is expensive and you will not send in a bee sample "just because", but rather to confirm your already promising preliminary wing measurements.





Определение пород пчёл,описания пород







apismelliferamellifera.0pk.me





Here found another good resource:





Анализ породности | Честная пчела ПЧЕЛИЧ|Пчеловодство| | VK


Вопросы Сергея Гурова




vk.com





More from Ukraine:





Методи визначення породи бджіл


Багатьох бджолярів цікавить яка порода бджіл є на власній або сусідській пасіці. Це хвилює і мене. Рік тому sij любязно надав мені зсилку на російську програму по обчисленню кубітального індексу (КІ), яку я використав та трохи переробив програму. Нижче подаю те що зробив та хочу почути думки інши...




tochok.info


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## msl (Sep 6, 2016)

GregV said:


> 18-point wing landmarking is a very skill-dimending and slow project


look at the tool that lets you set 4 point and then auto populate the rest 😉
Too much for a hobbyist, mabey


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## GregB (Dec 26, 2017)

msl said:


> look at the tool that lets you set 4 point and then auto populate the rest 😉
> Too much for a hobbyist, mabey


Give it a try and see for yourself.
LOL.

IMO, it does not pay to pursue the fine detail in pursuance for some endemic pure bees (in the US). They in the Euro keep looking for pure bees and this is where their focus is - to identify them! It is a valid project there - trying to restore some endemic bees best they can, for example. Even then, it is like stuffing the tooth paste back into the tube. 

We in US have a different problem (with no endemic bees to speak of).
This is on-going formation of totaly *new *hybrids with totally new signatures (the same process as the one in the Primorsky region). Some of these new hybrids are worthwhile keeping - but their patterns need to be identified first, even if roughly.


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## GregB (Dec 26, 2017)

MSL, anyhow, thanks for the YT find.
The video is literally few hours old and is worthwhile watching anyway because it is touching many of the same relevant points/topics.


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## Litsinger (Jun 14, 2018)

GregV:

I finally finished watching the two excellent BIBBA presentations by Dr. Tolfilski that MSL linked to. I also took the opportunity to follow-up with him to inquire as to what, if any benefit he saw in morphometric analysis of hybrid bees.

After a bit of back-and-forth, I came to understand that he asserts that locally-adapted bees will develop a unique, and fairly consistent morphometric 'fingerprint' after sufficient time of not being inundated by imports. So while this may not be of direct benefit to you right now, it did underscore a few points that have been batted around on this subject recently, namely:

1. It would be expected that the combination of genetic background and environment would lead to a morphometric signature in locally-adapted stock that likely could be utilized in evaluating the 'pedigree' of a particular population.

2. The specific environment that the particular bee stock is raised in has an impact on the morphometry. As such, even if genetic swapping is occurring between the the Primorski lines, the fact that they are being reared and maintained in different regions of the country will have an impact on the morphometric signature.

On a more practical level (at least for us in North America), Dr. Tolfilski referenced the work that Newfoundland is doing to protect their locally-adapted stock as a possible way forward for us if ever we decided to get serious about allowing local-adaptation to take hold:






CATCH THE BUZZ – Newfoundland and Labrador Bees are Special | Bee Culture







www.beeculture.com


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## GregB (Dec 26, 2017)

Hey Russ,
I am prepping the morpho-signatures of my 2020 stock (mostly dead now).
Unlike the Adam Tolfilski and his Russian/Ukrainian counterparts' goals, I am not really looking to isolate anything half-pure.
This is not possible with the bees here and the crudeness of my work anyway.
But a rough idea what our bees are about should be put out.

I read this comments about "pure" Saskatraz bees and I all can do is - cringe.
Any pure-ish bees should have documented half-honest morpho and genetic signatures.


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## GregB (Dec 26, 2017)

I got a hold of a sample of known "Russian" bees from a person that I know.

Indeed, the tools concluded this sample has significant prevalence of the A.m. sossimai signature (i.e. the Ukrainian bee signature).
Ukrainian bee is expected to be strongly predominant in the Primorsky/"Russian" bee.
If not so, I'd question the "Russian" label.


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## GregB (Dec 26, 2017)

One other thing I concluded - it is pretty much waste of time to be looking for any signs of A. m. mellifera in the generic North American samples.
I just excluded the AMM from my default reference sets.
Only time to look for potential AMM presence IF the Caucasian presence seems very strong and predominant, then AMM (M-lineage) is worth checking for (because signatures of O-lineage and M-lineage overlap outside of the C-lineage range).

For the most part all we have here is C-lineage and O-lineage (with C-lineage dominating).
Be great to find some samples where M-lineage is suspected (most likely some remote locations).


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## msl (Sep 6, 2016)

Hawaii?


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## GregB (Dec 26, 2017)

msl said:


> Hawaii?


I don't follow.


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## msl (Sep 6, 2016)

Mitochondrial DNA genetic diversity of honey bees, Apis mellifera, in Hawaii - Apidologie


Honey bees, Apis mellifera, in the Hawaiian Islands are geographically isolated from honey bees in mainland United States. We conducted a study on the mitochondrial DNA genetic diversity of honey bees from seven of the Hawaiian Islands by sequencing the intergenic region between the Cytochrome...




link.springer.com


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## GregB (Dec 26, 2017)

To compare, this VSH sample I have is predominantly Carnica with Ligustica flavor.


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## HaplozygousNut (Dec 30, 2015)

Here is an article of the bee genetics of America:








Genetic past, present, and future of the honey bee (Apis mellifera) in the United States of America - Apidologie


Humans have domesticated hundreds of animal and plant species for thousands of years. Artwork, archeological finds, recorded accounts, and other primary sources can provide glimpses into the historic management practices used over the course of a given species’ domestication history. Pairing...




link.springer.com





In this photo from the article it shows A. m. yemenitica genetics in New York. I didn't know about A. m. yemenitica being in the US:








Genetic past, present, and future of the honey bee (Apis mellifera) in the United States of America - Apidologie


Humans have domesticated hundreds of animal and plant species for thousands of years. Artwork, archeological finds, recorded accounts, and other primary sources can provide glimpses into the historic management practices used over the course of a given species’ domestication history. Pairing...




link.springer.com




New York had the whole genome looked at, which would be the nuclear DNA I think. The Mitochondria I have read are only the symbiotic bacteria in the cells of animals (the same as Chloroplasts in plants), and so their DNA is separate from the honeybee's DNA (Nuclear DNA). Although testing for mtDNA is probably a good way to see which subspecies of honeybees are in what areas, it is possible that it is not always accurate because the mitochondria of one subspecies of honeybee can be mixed into another subspecies of honeybee.

The M lineage in that photo in the article seems more common to the southern United States. I am thinking that Spanish Black bees are there in the Southern US. Spanish black bees are a mixture of M lineage.


The caption of the photo says:
_ "Overall, the figure highlights two major shortcomings in US honey bee genomics: small sample sizes (11 states have fewer than 11 samples) and biases towards either managed or feral colonies within states. Furthermore, despite the documented presence of AHBs in Louisiana, Alabama, Mississippi, and Georgia, sampling methods have missed them here. "_

I think that testing true feral colonies that have been isolated from commercial beekeepers would be much more diverse in subspecies.

Edit: Here is an interesting range map of the honeybee mtDNA: 








The South West Indian Ocean islands shelter a unique and novel African sub-lineage honeybee - BMC Series blog


Bees are very much in the news at the moment, with numbers declining due to factors such as climate change, pesticides and loss of habitat. However, it’s not all bad news; an article recently published in BMC Genetics describes the rather exciting discovery of a new African subgroup of bee...




blogs.biomedcentral.com





-Nathaniel Long


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## GregB (Dec 26, 2017)

Bump up.

This winter I have some other things to be spending my time on - not planning on doing the wing measurements.
But, if anyone has some kind of "black" bees (suggested AMM bees - to be clear), I'd be interested to work together and giving it a look.
I myself just have no such samples to bother with this work.

To be sure, the pure-enough M-lineage (i.e. the AMM bees) have the wing measurements distribution approaching this picture.









So far the best sample I got a hold was something suggesting high enough O-lineage (the Caucasians).
The bees sampled below were indeed very dark if to look at them superficially.

Be sure to catch that the blue on the above picture means M-lineage but on the picture below the blue means O-lineage (I simply twiked the program outputs).
Also to be understood - the wing measurements are no substitute for genetic testing (not even close).
It is rather to point out that the general looks and morphometric measurements should be at least consistent to make some kind of a educated suggestion about the specimens on hand.


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