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 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.