Scottish tracheal mite survey - Bees needed !

[emcampbell]

Hi everyone,

First a big thanks to all who I spoke with at the SBA centenary and a special thanks to Phil McAnespie and the organising committee – I had a brilliant time and it was great to see so many people with a shared interest in one place. Thanks to everyone who came and had a look at our research stall over the weekend as well - and for pointing out my mistakes !

The SBA and myself at the University of Aberdeen have recently (last week!!) been awarded a small amount of funding to develop a new method to diagnose small amounts of Acarapis woodi in honeybees that can be used by the bee inspectors and SASA. We are also going to try and find any link between tracheal mite and CPBV and get a very initial idea of tracheal mites spread across Scotland.

Phil and the SBA committee have worked with us before on a new approach to Varroa control and this is currently ongoing with some promising results. Many beekeepers have sent in drone brood and Varroa for this work and that has proven a great help to the project.

A few months ago Phil got in contact again after a new funding call was put out to our lab at Aberdeen Uni and mentioned that there may still be a problem with “tracheal mite” in Scotland and that anecdotal evidence suggests this is on the increase. We worked out a research plan and were fortunate to get the funding.
We now need Scottish beekeepers to help. We aim to survey 100 hives across Scotland initially. To do this we would ask beekeepers to get in touch as soon as possible with myself (e.m.campbell@abdn.ac.uk) and supply your postal address. I will then send out instructions on how to sample and send back bees as well as a vial of preservation liquid that will stop any DNA or RNA in the bees from degrading. The postage will all be covered by the project as will jiffy bags. All we need is your bees !

I have asked for between 20 and 30 bees from a hive as this should be enough to say with some degree of certainty that a hive has tracheal mite. I spoke with lots of far more experienced beekeepers than myself at the centenary and the conclusion was to use a matchbox or one of those wee boxes that paracetamol etc come in, opened about a cm and to run it over a frame of bees (being careful not to scoop up her majesty !!!). The bees can then be popped in the fridge or freezer for 5 mins to slow them down and then placed in the collection vial with preservation liquid I will supply. If you have a better way then feel free to do it your own way the only thing is to get the bees into the liquid either alive or just after they die.

I understand that many people will not be opening up their hives after late September / mid October dependant on weather so it would be great for people to get in touch as soon as possible so you can sample the bees before the shutdown. I had about 10 people register their interest at the centenary and have now sampled my own bees. If my maths is correct then only a few volunteers from each association would get me very near to the quota !

Thanks very much for your help,

Ewan Campbell

Post-doctoral research fellow
University of Aberdeen
School of Biological Sciences
e.m.campbell@abdn.ac.uk

[gavin]

Good to meet you on Sunday Ewan.

Ewan's leaflet is here (http://www.sbai.org.uk/misc/Trachealeaflet.pdf)in PDF form.

[Mellifera Crofter]

... The bees can then be popped in the fridge or freezer for 5 mins to slow them down and then placed in the collection vial with preservation liquid ...

Five minutes? I thought I've heard of bees coming alive again after even a day or so in a freezer - so five minutes - doesn't that mean drowning them?
Kitta

[gavin]

I believe that the idea is to slow the bees so that you can handle your sample of bees without them flying off, so that you can plonk them into the preservative solution. However, I doubt that 5 mins in a fridge is enough to do this, though that time in a freezer may be for a small sample of bees in a container that chills quickly. Maybe Ewan will comment. The study requires intact RNA which is a lot more unstable than DNA. Fully freezing and thawing the sample will probably destroy the RNA.

[marion.orca]

Why not use a Rennies box instead of paracetamol - they'd feel more at home. I'll email you, I'm happy for my bees to participate.

[Neils]

Sadly I'm a little too far "just" south of Aberdeen to participate, but I'm really happy to see this thread here for numerous reasons.

For those perhaps a little nervous of using small containers, though it is amazing just how many bees easily go into a small match box, an empty box of cooks matches is also worth considering though it is very easy to get far more than 30 in your sample.

[Jimbo]

Freezing and thawing will denature the RNA which is more unstable than DNA. I think Ewan is suggesting the cooling is so that you can work with the bees to get them as quickly as possible into the RNAse later. The RNAse later is a solution that protects the sample by destroying the RNAse enzymes that will break down the RNA

[Jon]

Five minutes? I thought I've heard of bees coming alive again after even a day or so in a freezer - so five minutes - doesn't that mean drowning them?
Kitta

I have heard that too but recently I collected some young bees to put in queen cages with virgin queens I had in an incubator.
I had the bright idea of immobilising them in the freezer and I put a plastic container with the bees in the freezer for 10 minutes.
When I took it out they were stone dead and I did try and revive them without any luck.

We have a PhD student from QUB starting a nosema survey which involves extracting DNA from samples.
I have to collect a sample of 50 older bees from 4 different colonies every month for him.
His instructions were to put the sample tube in the freezer and then cover them with ethanol once dead.
he even supplied a litre of ethanol but I have not made any cocktails with it yet.

[emcampbell]

Thanks for the interest everyone. Gavin is probably correct in that it is more likely to be 5 mins in the freezer not fridge.

Alternatively the bees can be put straight into the RNAlater but I have found it is easier to slow them down first. Unfortunately this does mean they will drown but it will keep the RNA / DNA intact. The freezing for 5 mins stage will no doubt make it a quicker demise.

A matchbox, paracetamol packet or rennies are all fine !

[Jon]

If you put bees in a small tightly closed container they suffocate before they freeze or drown.

[emcampbell]

I have had a great response so far including a number from on here. Still looking for more samples though especially from : Moray, Angus, Fife, Dumfries and Galloway, East Dunbartonshire, West Dunbartonshire, Renfrewshire, East Renfrewshire, North Lanarkshire, Falkirk, Clackmananshire, all the Lothians and Stirlingshire !

Keep them coming !

Cheers

Ewan

[The Drone Ranger]

I have heard that too but recently I collected some young bees to put in queen cages with virgin queens I had in an incubator.
I had the bright idea of immobilising them in the freezer and I put a plastic container with the bees in the freezer for 10 minutes.
When I took it out they were stone dead and I did try and revive them without any luck.

We have a PhD student from QUB starting a nosema survey which involves extracting DNA from samples.
I have to collect a sample of 50 older bees from 4 different colonies every month for him.
His instructions were to put the sample tube in the freezer and then cover them with ethanol once dead.
he even supplied a litre of ethanol but I have not made any cocktails with it yet.

Best get some hardy hybrids I put mine in a freezer for 24 hours then took them to a microscopy course at Stirling uni.
As the course progressed more and more of them revived and started flying around the room
All they needed was an apidea and a queen and they would have set up home in the lab

[Mellifera Crofter]

... especially from : Moray, Angus, Fife, Dumfries and Galloway, East Dunbartonshire, West Dunbartonshire, Renfrewshire, East Renfrewshire, North Lanarkshire, Falkirk, Clackmananshire, all the Lothians and Stirlingshire !

Keep them coming !

Cheers

Ewan

I'm glad Aberdeenshire doesn't feature in your list. It makes me feel a little bit less guilty for not having offered to send you a sample yet - particularly after Drone Ranger's post.
Kitta

[Jon]

Best get some hardy hybrids

LOL
That's heterosis for you. Spawn of the devil near impossible to kill. Must be like a zombie movie in your apiary.

Out of the woodwork or awakened from his crypt, the Drone ranger rides again.
I was looking out for Silver in the car park at the Scottish centenary but you obviously had a more pressing engagement.

[The Drone Ranger]

LOL
That's heterosis for you. Spawn of the devil near impossible to kill. Must be like a zombie movie in your apiary.

Out of the woodwork or awakened from his crypt, the Drone ranger rides again.
I was looking out for Silver in the car park at the Scottish centenary but you obviously had a more pressing engagement.

Busy time of year for beekeeping might log in more during winter but I don't have much more to say without just repeating myself.
Re Centenary I am not a member of SBA now so could spend £30 down the pub instead :)
Still check out the forum to see what people are saying but I don't post often these days.
Wonder how many tracheal mites have survived thymol treatments interesting stuff

I use IKF from brunel microscopes on the rare occasions when I want to kill a bee or two before taking off their head or abdomen etc.
But it feels wrong -- Bad Karma Tonto

[gavin]

There was a [name deleted in the interests of privacy, but it was only a letter or two different from DL's] in attendance and sporting a name badge. Jon's blurry vision after the excesses of the night before got him excited for a while but there was no Tonto to be seen anywhere so I was always suspicious.

All in the name of science, DL. The freezer treatment just subdues them so that they can meet a watery end in a molecular biology reagant. 20-30 bees in a matchbox should slow down significantly in that time. Yes, I too have had bees coming into life after 24 hrs in a freezer. Bigger numbers of bees in bigger containers can fight the cold for a while, poor souls.

I have my package from Ewan and it is a nice day so maybe I'll be trying it out today if I can find a matchbox or a Rennies or something similar.

Get your request in Kitta - Ewan needs the numbers, especially as tracheal mite isn't that common. Yes, OK, miticides might have cleared it out from much of the East but there could be miticide-resistant types around, or maybe they are not as susceptible as Varroa to some of the treatments. I almost started more east-west speculation there but thought better of it.

[marion.orca]

That's it - dastardly deed completed with a heavy heart, but needs must sometimes. I used a plastic container and just swept some off the entrance where they were nicely warming up in the sunshine, only to go into a cold dark freezer. I got 41 in total, so have put the remaining 11 into the sunshine to see if they possess the same remarkable traits that DR's have after an hour being frozen - I somehow doubt it, but we'll see.

[gavin]

First of my two lots done - I'm doing one set of my own and one from the association apiary. Little cardboard Tesco aspirin box, so it ought to cool quickly in the freezer. They were still walking about and buzzing after 15 min but had gone quiet and looked dead at 40 min. The extra ones started coming back to life after 20 min warming up again.

Bit of a squeeze getting the sample into the tube but with a bit of shaking they ended up more or less submerged.

[Mellifera Crofter]

That was probably not the way to get me to do the 'dastardly deed', Gavin ... Ouch.
Kitta

[marion.orca]

My spare 11 that made it to the freezer, but not the tube, are well and truly goners. So no remarkable powers in my bees. That made me feel really bad after reading Gavins post of his coming back to life, so to rid myself of my guilty feelings, I rescued a Hover fly from drowning in the water butt and gave it a drop of syrup - am I redeemed in the insect world ?

[emcampbell]

Hey Marion. I have to say mine were pretty much goners after about tens mins in the freezer - I might try reducing that time to five mins. Althought it was easy to get them into the sample liquid after the freezer. Whats your address Marion ? PM me.

[marion.orca]

I added 2 or 3 at once, gave it a good shake, and carried on like that until all 30 were submerged.

[Eddie]

Hey, got my kit to take the samples which I will do, as soon as this rain stops and the sun shines for about 10 minutes..

[gavin]

Took the second sample in the easterly gale at lunchtime and gave them 30 min to cool down this time. Three were still buzzing about in the box (and were the spares shaken out near the apiary on my way back to work). Well on their way to Aberdeen by now.

[Trog]

My sample was dead after 15 mins in the freezer and the spare 5 didn't revive in the kitchen. Should be there by now - just as well they left the island yesterday!

[EmsE]

I'm not too sure what type of bee I have- they've been on the freezer for 20 min and still too active to get them into the tube. We've got 4 in already and about the same number now flying about the room. Must be that kilbarchan vigour:rolleyes:

[EmsE]

All in now, after 25 min in the freezer, but those that went in looking dead started moving around again in the liquid so will drown:( a few left in the freezer that will be heading for under the microscope at some point?

[nemphlar]

15 mins in the freezer did for mine,no sign when stuffing them in the tube. I'd be checking the operating temperature of my freezer

[emcampbell]

Hi again everyone,
I have had a great response with the odd enquiry still coming in. I have sent out about 60+ sampling tubes with very good responses from some areas and patchy to none in others. It would be really good to get as large a coverage as possible so if anyone from Fife, Angus, Lothian, Lanarkshire and anywhere between Moffat east to Stranraer would still like to get involved then that would be a big help (and elsewhere!). Any hive is a usefull sample - no need to have suspected tracheal mite or anything else !
Cheers
Ewan

[gavin]

I'll be introducing the SBA touring speaker - Richard Ball - at our meeting in Dundee on Friday. I'll remind the audience and encourage some more to send you samples.

[marion.orca]

Is it still too early to see results for this survey ? and will we all get individual results or a more general conclusion ?

[gavin]

Good questions. I know that he has some tentative conclusions from what he's seen so far, but this probably isn't the place to spill the beans. Drop him a line and he may be able to tell you something.

[marion.orca]

Thanks Gavin. Ewan has replied to my questions.

[emcampbell]

Hi everyone,

I just wanted to let you all know that I have so far done about 50% of the hives. We are getting through a fair number each week but I am still doing all the Varroa work and getting ready for the next phase of that.... so its been slower than we hoped ! Another thing is that we have found quite a high percentage of hives do show positives for Acarine. More than I thought ! So I am currently double and triple checking all the data. I hope to get questionnaires out to everyone by email or post next week - I delayed this due to the long winter as I figured some of you may not have checked hives yet.

Thanks !

Ewan

[The Drone Ranger]

wonder if the AMM samples have more or less acarine.
(they will bee the black ones with no wings )

[Jon]

I would say you are right. Some of them will have more and some of them will have less!

[Jimbo]

we are already ahead of you DR. Ewan sent me the first 12 samples for wing morphometry as part of a survey I am doing to see where the Amm are located in Scotland. I will send Ewan back the results so that he can check to see if there is any correlation between the sub species and Acarine

[Jon]

he can check to see if there is any correlation between the sub species and Acarine

That's shill talk Jimbo. Looking at a bee health issue which has nothing to do with pesticides!

Acarine was a big issue in my neck of the woods in the 1970s.
It is still about though and I am going to get some samples tested shortly as a lot of beekeepers are reporting higher losses than usual and we need to rule in or out stuff like tracheal mites and nosema.

[The Drone Ranger]

we are already ahead of you DR. Ewan sent me the first 12 samples for wing morphometry as part of a survey I am doing to see where the Amm are located in Scotland. I will send Ewan back the results so that he can check to see if there is any correlation between the sub species and Acarine

You mean some of the bees he received still had wings on them :)
They must be Hybrids

[Trog]

we are already ahead of you DR. Ewan sent me the first 12 samples for wing morphometry as part of a survey I am doing to see where the Amm are located in Scotland. I will send Ewan back the results so that he can check to see if there is any correlation between the sub species and Acarine

That will be very interesting indeed. THEN all we need is someone to come up with a good cure for it.

[The Drone Ranger]

That will be very interesting indeed. THEN all we need is someone to come up with a good cure for it.

Cure Wing morphometry ??
That's an addiction could be tough

[Trog]

:D Very good, Droney!

[The Drone Ranger]

http://www.youtube.com/watch?v=6wy2PG_MB4Y
little video from states about dissecting and control

[Trog]

Very interesting. Maybe I'll try making up a patty or two! The dissection seemed a bit of a palaver, though.

[The Drone Ranger]

Very interesting. Maybe I'll try making up a patty or two! The dissection seemed a bit of a palaver, though.

I think your right I just pull the head off and the collar but if you have never done it its quite a good explanation