Category Archives: Varroa control

Apivar & Apitraz = Amitraz

The range of miticides available ‘off the shelf‘ to UK beekeepers has recently been increased by the introduction of Apitraz and Apivar.

‘Off the shelf’ because, until recently, these were only available with a veterinary prescription.

Considering the extensive coverage on this site of oxalic acid-containing miticides and more recent posts about the – regularly ineffective – Apistan, it seemed fair and appropriate to write something on the active ingredient and mode of action of these new products.

Mites on drone pupae ...

Mites on drone pupae …

Conveniently, because the active ingredient is identical, these can be dealt with together in a single post. The similarities don’t end there. The amount of the active ingredient is the same and the way it is administered is very similar. They are different commercial products; Apitraz is distributed by Laboratorios Calier, SA and sold by BS Honeybees, Amitraz is distributed by Veto Pharma and sold by Thorne’s. The strips have a different appearance and a slightly different mechanism by which they are hung in the hive.

They even cost about the same – a single packet of 10 strips (sufficient to treat 5 hives) costs £30.50 and £31 respectively for Apitraz and Apivar.

Amitraz

The active ingredient in both Apitraz and Apivar is Amitraz.

Yes … I find these three names confusing similar as well 😉

Amitraz is a synthetic acaricide – a pesticide that kills mites and ticks. It was discovered and developed almost 50 years ago by the Boots Co. (the drug development predecessor of the Boots the Chemist 1 found in most high streets). Amitraz is the active ingredient in a range of medicines approved by the Veterinary Medicine Directorate, including Aludex and Certifect, both of which are used to treat mange in dogs.

Amitraz

Amitraz …

For completeness I should add that Amitraz used to be used by US beekeepers and was sold as a generic pesticide under the name Taktic, though this was withdrawn in about 2014. I believe that Apivar is now available as a slow-release Amitraz-containing Varroa treatment in the US.

Mechanism of action

Amitraz has to be metabolised (essentially ‘modified’) before it is active. This modification occurs much less well in bees than in mites. In fact, the toxicity of Amitraz for bees has been determined to be about 7000 times less than in mites.

Once converted into an ‘active’ form the most important mechanism of action for Amitraz is through interaction with the alpha-adrenoreceptor and octopamine receptors of Varroa 2.

OK, since you asked … octopamine receptors normally bind a neurotransmitter called – rather unimaginatively – octopamine. Quelle surprise as an apiculteur would say. It’s likely that occupancy of these receptors by Amitraz triggers a series of so-called downstream events that change the behaviour of Varroa. Similarly, amitraz also acts as an agonist 3 when binding to the alpha-adrenoreceptor which normally interacts with catecholamines. This results in neurotoxicity and preconvulsant effects.

That all sounds a bit vague. Essentially, amitraz binds and activates receptors that are critically important in a range of important aspects of the Varroa activity and behaviour. Remember here that the mite is entirely dependent upon proper interaction with the bee to complete the life cycle. For example, if the mite fails to enter a cell at the correct time or doesn’t hitch a ride on a passing nurse bee for a few days, it will likely perish.

Amitraz changes behaviour and so exhibits miticidal activity. It has additional activities as well … these multiple routes of action may explain why resistance to amitraz is slow to develop. More on this later.

Usage of Apitraz and Apivar

Both Apitraz and Apivar are formulated as plastic strips impregnated with amitraz. The bees must come into contact with the strips to transmit the amitraz around the hive. Two strips are therefore placed between frames approximately one-third of the way in from each side of the brood box – typically between frames 4 & 5 and 7 & 8 of an 11 frame box. This assumes the bees occupy the entire box. If they don’t, arrange the strips in the appropriate part of the box with 2 frames separating them. Both types of amitraz-containing strips have a means of securing them hanging between the frames.

The recommended treatment period is 6 (Apitraz, or Apivar with little/brood present) to 10 weeks (Apivar with brood present). As with Apistan, treatment should not be applied during a honey flow or when honey supers are present. Further details are included on the comprehensive instructions provided with both products. There’s also a reasonable amount of information on this New Zealand website for Apivar.

Efficacy

This is the good bit … very, very effective. When used properly, amitraz-containing miticides can kill up to 99% of the Varroa in a colony.

Toxicity and wax residues

The good news first. Amitraz does not accumulate in wax to any significant extent. It is not wax-soluble. This is in contrast to Apistan which is found as a contaminant in most commercially-available beeswax foundation.

And now the bad news. Beekeepers also have alpha-adrenoreceptors and octopamine receptors. So do dogs and fish and bees. Although amitraz has increased specificity for the receptors in mites and ticks, it can also interact with the receptors in other organisms. Consequently, amitraz can be toxic. In fact, if you ingest enough it can be very toxic. Symptoms of amitraz intoxication include CNS depression, respiratory failure, miosis, hypothermia, hyperglycemia, loss of consciousness, vomiting and bradycardia.

And it can kill you.

Admittedly, the doses required to achieve this are large, but it’s worth being aware of what you’re dealing with. Amitraz-containing strips should be used only as described in the instructions for use, handled with gloves and discarded responsibly after use.

Resistance

Multiple modes of action makes it much more difficult for resistance to evolve. But it can and does. Resistance to amitraz is well-documented and is understood at the molecular level. However, this is in cattle ticks, not Varroa.

At least, not yet, though there are numerous anecdotal reports of Varroa resistance.

I’ll deal with resistance in a separate post. It’s an important subject and avoiding it is a priority if amitraz-containing compounds are going to remain effective for Varroa control.

Cost

At about £6 per colony, amitraz-containing treatments are not significantly more expensive than the majority of other approved miticides, perhaps with the exception of Api-Bioxal which is appreciably less expensive (though more restricted in the ways it can effectively be administered 4).

Apivar ...

Apivar …

When you purchase a couple of packets of Apivar – enough for 10 colonies – it might feel expensive 5. However, it’s worth remembering that this is still less than the likely ‘profit’ on a couple of jars of your fabulous local honey per colony per year, which seems pretty reasonable in the overall scheme of things.

And, if you look after your colonies well, you are maximising the potential yield of honey in the future … so you’ll be able to afford it 😉


 

Weather to treat

Not Whether to treat? … to which the answer is yes. Instead, a poor pun on the choice of how I use temperature as an indication of when to treat colonies in midwinter …

Midwinter OA-based treatments

Oxalic acid-based treatments for midwinter Varroa control are most effective when colonies are broodless. This is because oxalic acid (OA) treatments only kill phoretic mites and are ineffective against mites in sealed cells. They are therefore ideal for use on swarms, packages and broodless colonies in midwinter.

These OA treatments include Api-Bioxal, the VMD-approved treatment, and unmodified oxalic acid, it’s active ingredient. The importance of midwinter treatments, the preparation of the OA solution and how to trickle treat have recently been covered. I’ve previously discussed sublimation and will do so again in a longer article in the future.

The beekeepers winter dilemma

How can you tell whether your colonies are broodless in midwinter?

On a warm, sunny, Spring afternoon this takes just a couple of minutes … remove the roof, crack off the crownboard, gently lift out the dummy board and the adjacent frame, look carefully at the mass of bees covering the top bars, aim for about the middle and gently prise apart those two frames, lift out a frame from one side of the ‘gap’ and – Hey presto – brood.

Just writing that in early December makes me hanker for much warmer days …

Memories of midseason

Memories of midseason

Actually, you can do exactly the same in midwinter. There are videos on the internet showing an experienced and (in)famous Finnish beekeeper opening his colonies at -10ºC.

I’ve opened and briefly inspected colonies at low temperatures (though not sub-zero). The bees are usually pretty torpid, reluctant to fly – or simply too cold to – and you can be in and out in just a minute or so. Bees cope pretty well with this. It undoubtedly disturbs them a bit and it breaks the propolis seal on the crownboard, but – done carefully and quickly – it’s the only foolproof way to determine whether a colony is broodless in midwinter.

But what if they’ve got brood and it’s therefore not the optimal time to treat? Do you go back and repeat the entire process in 1-2 weeks? What if it’s snowing next time, or there’s a howling gale blowing?

An alternative approach is needed.

The annual brood rearing cycle

As the colony moves from summer to autumn the egg laying rate of the queen drops. It goes on dropping, although not necessarily smoothly, as the days shorten further, the temperature drops and the sources of pollen and nectar disappear. If the queen stops laying altogether then the colony will become broodless about 21 days later.

At some point, perhaps early in the New Year, the queen starts laying again. Slowly at first, but at increasing levels as the season starts. Once foraging starts in earnest the egg laying rate increases markedly and peaks sometime in June.

The precise timing of all these changes cannot be predicted. It’s likely to be dependent on a range of factors – nectar and pollen availability, the strain of bee, day length (and whether it’s increasing or decreasing) and temperature.

Of these, temperature probably has the greatest influence.

Probablyß.

Generalised annual brood and worker numbers ...

Generalised annual brood and worker numbers …

Here’s a quick’n’dirty graph put together with BEEHAVE showing a generalised annual cycle of total brood (blue) and adult bee (red) numbers. Under the conditions in this model the colony is broodless for ~30 days at the end if the year.

Temperate(ure)

Part of the problem with being definitive about the annual brood cycle is the temperature variation with latitude. Temperate regions stretch – in Europe – from Northern Finland to Southern Spain. Bees are kept throughout this range, but obviously experience wildly different climates.

And then there’s the year to year variation.

So if you can’t predict when the colony is going to be broodless, perhaps you can observe the weather – and in particular the temperature – and make an educated guess.

Watch the weather

Over the last few years I’ve applied my midwinter treatment soon (<6 days) after the end of the first extended cold period of the season. This is generally earlier than most beekeepers, who often treat between Christmas and New Year, or early in January.

So, how do we reasonably accurately monitor the weather for a suitable time to treat?

Ho ho ho

Ho ho ho

Most of us live in centrally-heated splendour, protected from the day-to-variation of temperature by heated car seats, air conditioning, hot water bottles, Thinsulate and wood-burning stoves. Do you know what the temperature was today? Rather than trust the wildly-variable (in accuracy) national weather reports for the actual temperature near my apiaries, I instead use very much more local data from Weather Underground.

There are hundreds of ‘amateur’ weather stations across the country that upload data to wunderground.com. Most of these provide current and historic data, including temperature (max, min and average). Here’s one for Auchtermuchty in Fife (on wunderground.com) and directly from the weather station.

Once the weather cools I keep an eye on the average temperature over an extended period of a fortnight or so. If it remains low I wait a bit more … but I then treat as soon as practical after it warms up to 8-10°C or so.

The proof of the pudding

Here’s a graph of the temperature data for 2016§. As indicated on the graph, I treated colonies on the 7th of December.

2016 temperature data and OA treatment ...

2016 temperature data and OA treatment …

I didn’t open my colonies, but others opened on the same day nearby were all broodless. The 7th was chosen as it was the first warm (relatively!) day after a 19 day window in which the average temperature had barely climbed above 5°C.

These treated colonies went into the New Year with vanishingly low Varroa levels.

And again …

This year appears to be repeating a very similar pattern. We’ve had frosts most nights since the 10th of November. It started to warm up significantly in early December as storm Caroline bore down on Scotland and I treated most of my colonies on the 6th 

… by the light of a head torch, in light rain and strengthening wing at 7pm after work.

No, I didn’t open any of the hives to check if they were broodless  😉

It was over 11°C in the apiary when I treated, the barometer was plummeting and the forecast was for near-zero temperatures within 24 hours and remaining that way for another 10 days.

Some of my hives have perspex crownboards. These allow me to check both the state of the colony and if the vapour from my Sublimox has permeated to every corner of the hive. All the colonies were very loosely clustered, with a few bees even wandering out briefly onto the landing board in the dark as I bumbled around preparing things.

The Varroa trays will now be checked in a week or so to work out the mite infestation levels. In the meantime, I can start planning for the coming season knowing I’ve done the best I can to reduce virus levels in the colonies, so giving them a good start to the year.

A Hi tech solution?

Colonies rearing brood maintain a higher, and stable, broodnest temperature (32-35°C) than colonies without brood. It is therefore possible to determine whether a colony has brood by monitoring the temperature directly, rather than trying to infer it from the ambient temperature.

Brood rearing starts ...

Brood rearing starts …

Arnia make hive monitors that allow this sort of thing to be measured. It would be interesting to relate the brood temperature to the ambient temperature (described above) to determine how accurate or otherwise simply ‘watching the weather’ is. Of course … what you’d really want to do is monitor when brood rearing stops and treat soon after that.

Stop press

I treated colonies in our research apiary the following day – the 7th – with dribbled Api-Bioxal. The temperature had dropped almost 7°C since the previous evening and colonies were again beginning to cluster tightly. Under these conditions I’m never confident that the OA vapour penetrates fully, so prefer to trickle treat.

I briefly checked one strong colony in a poly hive for brood.

It was broodless, as I’d hoped  🙂

Of course, this doesn’t guarantee all the others are also broodless, but it does give me some confidence that I’d chosen the correct weather to treat.


† This article, like most on this site, discuss beekeeping issues relevant to temperate climates. It’s important to make this clear now as most of what follows is irrelevant to readers from warmer regions.

∞ Even if there is brood in midwinter, it’s going to be in pretty small amounts. The rate at which this brood emerges is going to be low. The chances of determining what’s going in the colony by ‘reading the tea leaves’ from the debris falling through the mesh floor of the hive is therefore not great. It would probably also require repeated visits to the apiary.

ß This needs qualifying … in midseason, when the temperature varies but it’s not generally cold, the nectar flow is probably the rate-limiting step for brood rearing. The June gap is regularly associated with the queen shutting up shop for a while. However, in late autumn and early winter I’m sure the plummeting temperatures is a major influence on egg laying by the queen.

‡ National … Ha! Most are only national if you live within the M25. Anywhere else and you’re usually much better off accessing some data from closer to home. It’s worth noting that the sort of ‘amateur’ weather stations I discuss do vary in data quality. For example, they’re a bit dodgy recording temperatures in full sun (they tend to over-read). However, if you find a local one, check the temperature in comparison to a thermometer in your apiary, you’ll find it’s a useful way to monitor what might be happening in the hives.

§ I don’t routinely generate these graphs – I have a life (!) – but did specifically to illustrate this post. It’s sufficient to simply monitor the average temperature.

Colophon

Whether the weather be fine
Or whether the weather be not,
Whether the weather be cold
Or whether the weather be hot,
We’ll weather the weather
Whatever the weather,
Whether we like it or not.

Anonymous

 

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Trick(le) and treat

Tools of the trade

Tools of the trade

This is the third and final post on why, with what, when and how to minimise mite levels in colonies in midwinter.

In the first post I explained why midwinter mite treatment makes sense. In the second I described how oxalic acid-containing solutions should be prepared and stored.

Oxalic acid-containing” solutions includes both Api-Bioxal, the VMD approved treatment, and the unadulterated chemical. All three posts focus on trickling or dribbling – I’ve covered sublimation previously and both are essentially equally effective. Sublimation or vaporisation is currently very fashionable … but trickling is simplicity-itself and requires almost no special equipment.

In this post I’ll discuss how to administer the oxalic acid-containing solution.

For readability I’ll use the term OA solution to mean any oxalic acid-containing solution. About 50% of the readers of this site are from outside the UK; local rules may determine what you are or are not allowed to administer to your bees.

Trickling or dribbling

You’ll hear both terms used interchangeably1. The general principle is that you directly administer 5ml of a 3.2% w/v solution of oxalic acid in thin (1:1) syrup per seam of bees in the colony.

Directly‘ because you administer the OA solution to the seam of bees. You don’t count the seams and then simply pour it into the hive. You don’t spread it across the top bars. The idea is that the bees at the top of the seam get coated in the solution and that it dribbles down through the colony, being passed from bee to bee as they feed and groom and move about.

Two seams of bees

Two seams of bees …

During this process any phoretic mites will also get exposed to the oxalic acid. Since mites are readily damaged by the OA solution they fall off and gradually drop out of the bottom of the cluster. Gradually, as it takes a few days for gravity to deliver all the corpses.

You can therefore determine whether mites were present and killed by placing a Varroa tray underneath the open mesh floor of the hive. Note that this doesn’t tell you how effective the treatment has been … for that you’d need to know the mite infestation level before treatment as well.

When to treat

In many ways this is the critical decision. As described previously, maximal benefit occurs when the colony is broodless. Ideally you want an extended cold period late in the calendar year. The colony will cluster tightly and brood rearing will slow down or stop completely.

If the cold period has lasted 2-3 weeks, even better. This will mean that some or all of the brood present will have emerged. The more sealed brood present, the less effective trickling OA solution is as a means of controlling mites.

Choose a calm, cool or cold day. I usually wait for a day with temperatures between 0 and 5°C. Much warmer than that and the cluster starts to break up or the bees are more likely to fly about as the crownboard is lifted. Windy or wet days disturb the bees (at least when you prise the crownboard off), so it’s best to avoid those.

I prefer to treat before the year end, rather than after, if I can. From a few irregular midwinter peeks into the cluster I think queens start laying earlier than most beekeepers think.

It pays to be prepared …

Trickle 2 - £1

Trickle 2 – £1

… Aesop (~620-560BC) was right, though he wasn’t talking about beekeeping. Before treating your colonies there is some preparation needed. Do this properly and it’s a doddle.

Purchase a Trickle 2 container from Thorne’s. These are a measly quid each. You’ll only need one.

Practice with the Trickle 2 container (see below).

Gently warm your pre-prepared OA solution to about 25°C. If you made it up in advance and stored it at 4°C in the fridge this will take an hour or two. The easiest way is to stand the container (preferably thin-walled … I use a well-rinsed milk carton) in a basin of warm water.

Pour the pre-warmed OA solution into a well-labelled vacuum flask. You can buy these from Tesco for £2.50 with a capacity of 1 litre. The aim here is to take everything you need ready-prepared to the apiary so the treatments take the minimum time possible.

Remember that OA is toxic. Label everything carefully, make sure children can’t get near it and don’t use it again for food/drink purposes.

That’s it … you’re ready. You’ll need a hive tool, a bee suit, thin gloves (to protect you from the OA, not the bees), your vacuum flask of OA solution and the Trickle 2 bottle. By all means take your smoker, but you shouldn’t need it.

I’ve got a 5 ml (or 25 ml) syringe … won’t that do?

Yes … but no.

A Trickle 2 bottle holds 100ml of prepared OA solution. It takes two hands to fill the bottle, but only one hand to use it. That 100ml is sufficient for 20 seams of bees i.e. two completely full colonies (assuming an 11 frame National box). In midwinter the colony is unlikely to occupy 10 seams. A Trickle 2 bottle is also pretty accurate, reproducibly dispensing about 4.6-4.8ml of liquid. That’s close enough to 5ml.

In contrast, a syringe also takes two hands to fill (and refill). However, unless it’s a 5ml syringe, it’s difficult to accurately and reproducibly dispense liquid without using two hands. A 5ml syringe gives you the necessary accuracy, but needs refilling for every seam of bees. This takes time … during which the crownboard is off and the colony is getting chilled.

I’ve done both and can assure you that the Trickle 2 bottle is much better. Just buy one. It’s only £1 and it’ll last ages if one of your association members doesn’t borrow it … or doesn’t return it.

How to use a Trickle 2 bottle

  • Remove the cap and fill to the top of the lower chamber with liquid (practice with water).
  • Replace the cap.
  • Hold the bottle with your thumb and fingers on opposite sides of the lower chamber, with the external ‘pipe’ to the upper chamber next to your palm.
  • Undo the spout about a turn.
  • Gently squeeze the lower chamber. Liquid is forced up the pipe into the upper chamber. Hold it against the light to observe this.
  • Once the upper chamber is full, stop squeezing. Excess liquid drains back into the lower chamber.
  • If you are right handed turn the Trickle 2 bottle anti-clockwise2 using your wrist and gently squeeze the bottle to dispense the liquid in the upper chamber from the spout. If you’re left handed you need to turn the bottle clockwise.

And in practice

The single-handed operation for the Trickle 2 container really pays dividends when treating a colony. You can gently prize up one side of the crownboard, hold it in one hand, administer the OA solution to each seam with the other hand and gently lower the crownboard back down … all in less time than it took me to write that.

Like this:

This is a reasonably sized colony being treated in the second week of January 3 years ago. The video is 1’45” long, but the crownboard is only open for about 50 seconds. And I was chatting with Mick Smith off camera, so could have perhaps gone a bit faster if I’d concentrated … 😉

Here’s a more detailed view of treating a small colony:

33 seconds of warmed, acidic goodness to slaughter the mites and give the colony the best possible start to the upcoming season.

Cautions and considerations

Discard any OA solution that’s not been used. Warming it will have raised the HMF levels and this may be toxic for your bees. However, read footnote 3 for another way to avoid HMF buildup3.

Wash everything carefully – the Trickle 2 bottle, the vacuum flask, gloves etc. Since the OA solution was in syrup everything gets sticky and gummed up. Clean stuff up, make sure it’s labelled and not going to be used in the kitchen and put it away until next year.

Oxalic acid kills mites, but it’s also toxic for unsealed brood. This is perhaps unsurprising considering it has a pH of 1 (i.e. very acidic) and ‘naked’ larvae aren’t protected by the tough exoskeleton that adult bees have. This is another reason to treat during a broodless period in midwinter.

In summer, swarms can also be treated with trickled oxalic acid-containing solutions before they have sealed brood. If a swarm arrives in bait hive, let it settle and start drawing comb on the foundationless frames. A day or so later treat it with oxalic acid by trickling. When I’ve done this I usually wait until late afternoon or early evening, so most of the bees are in the box. The colony obviously won’t be clustered, but the principle is the same – 5ml of syrup down each seam. Easy peasy. Effective.

Swarms have a significant mite load, so it’s well worth treating them before they rear brood and give the phoretic mites somewhere to breed.

Finally, it’s often recommended that a colony is only treated once per year with oxalic acid by trickling or dribbling. I’m not sure where this advice originates, but it’s probably wise.

‘Vaping’ vs. trickling

The discussion forums are awash with recommendations to ‘vape’ the colony, rather than trickle. Vaporisation, or more correctly sublimation, is a widely used method and has been in use for two decades. It’s currently very fashionable. I’ll write a more substantial comparison sometime in the future, but the following brief notes might be of interest.

Sublimation can be done repeatedly with brood present (though there’s no peer-reviewed evidence of efficacy) and is both well-tolerated by the colony and is not toxic to unsealed brood. It requires specialised and potentially expensive equipment, both for delivery and personal protection. You can build your own vaporiser, but shouldn’t skimp on protection for the operator. With a well designed vaporiser and hive there’s no need to open the colony to administer treatment.

In contrast, trickling requires only the Trickle 2 bottle and vacuum flask described here. Personal protection is a pair of latex gloves. It should only be conducted when the colony is broodless, should probably only be conducted once and does require the hive to be opened (albeit briefly).

You’ll be told that vaporisation is faster. It isn’t. Watch the videos above. Even my Sublimox – probably the fastest ‘active’ vaporiser on the market – takes well over a minute per colony if you take into account sealing the box, moving the generator about, unsealing the hive etc.

There are reports that sublimation is more effective, but the difference is marginal, and possibly not statistically significant. There is also a report that colonies are stronger in the Spring after sublimation, though this may be due to toxicity to open brood by trickled OA solution. If the colony is broodless this shouldn’t be an issue.

I’ve used both many, many times without a problem. Across the UK I suspect more beekeepers trickle OA, rather than ‘vape’ (a word I dislike), though the vocal ones on the discussion forums currently favour vaporisation.

What’s more important than how you deliver the oxalic acid, is that you do treat. Trickling OA solution is so easy and inexpensive that there’s no reason not to … and your colonies will be much healthier for it.

Get dribbling 😉


If the beekeeper is of a certain age you’ll hear these terms used in a different context. We’re restricting discussions here to delivering OA 😉

If you are left handed you need to turn the Trickle 2 bottle clockwise. Actually, to be pedantic, if you are left handed and holding the bottle in your left hand, turn it clockwise. It’ll make sense once you try.

3 In the previous article on preparing oxalic acid solutions Calum posted a comment on preparing the OA in water and only adding and dissolving the required amount of sugar just before use. This has the advantage that there will be no HMF buildup. OA solution in water should be perfectly stable. I’ve not done it this way, but it makes sense and might be worth trying.

Colophon

The title of this article is a twist on the term Trick or treat. This is not entirely inappropriate as Trick or treating is a Halloween (31st October … just a few days away) custom dating back – in various forms – centuries.

The modern usage, essentially North American, dates back to the 1920’s and refers to children in costumes going house to house threatening to play a trick unless the homeowner provides a treat, usually sweets or toys. In Britain these traditions date back to the 16th Century, both of children going house-to-house asking for food and of dressing up in costumes at Halloween.

Closer to home, ‘guising‘ – children in Scotland going from door to door in disguise asking for food, coins or chocolate  – dates back at least a century.

The term Trick or treat only entered common usage in the UK in the 1980’s.

 

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Oxalic acid preparation

This is the second of three articles on midwinter treatment of colonies with oxalic acid to minimise Varroa levels. In a recent post I explained why a midwinter treatment was necessary, even if you’d treated three months earlier. Essentially this is because:

  • there will still be some residual Varroa, particularly if you treated in late summer rather than early autumn (and this post explains why early treatment is preferable)
  • midwinter is the time when brood levels are at a minimum, so most mites will be phoretic and readily accessible to the miticide treatment

Midwinter is the time to use oxalic acid-containing treatments. It can be delivered in a variety of ways; by sublimation (vaporisation), spraying or trickling (dribbling).

Trickling or dribbling

This post is about the preparation and storage of oxalic acid-containing solutions for trickling. Sublimation is covered elsewhere and spraying is not approved or widely used in the UK.

The process for trickling is very straightforward. You simply trickle a specific strength oxalic acid solution in thin syrup over the bees in the hive. The oxalic acid kills the mites. How isn’t entirely clear – it’s thought to corrode the mouthparts and soft tissue. It’s more than 90% effective in killing phoretic mites when used like this.

Beekeepers have used oxalic acid for years as a ‘hive cleaner’, as recommended by the BBKA and a range of other official and semi-official organisations. All that changed when Api-Bioxal was licensed for use by the Veterinary Medicines Directorate (VMD).

Oxalic acid and Api-Bioxal, the same but different

Spot the difference ...

Spot the difference …

Api-Bioxal is the VMD-approved oxalic acid-containing miticide. It is widely available, relatively inexpensive (when compared to other VMD-approved miticides) and very easy to use.

It’s very expensive when compared to oxalic acid purchased in bulk.

Both work equally well as both contain exactly the same active ingredient. Oxalic acid.

Api-Bioxal has other stuff in it (meaning the oxalic acid content is a fraction below 90% by weight) which actually makes it much less suitable for sublimation.

How much and how strong?

To trickle or dribble oxalic acid-containing solutions you’ll need to prepare it at home, store it appropriately and administer it correctly.

I’ll deal with how it is administered next time. This is all about preparation.

The how much is easy. You’ll need 5ml of oxalic acid-containing solution per seam of bees. In midwinter the colony will be reasonably well clustered and its likely there will be a maximum of only 8 or 9 seams of bees, even in a very strong colony.

Hold on … what’s a seam of bees?

Two seams of bees

Two seams of bees …

Looking down on the colony from above, a seam of bees is the row visible between the top bars of the frames.

Remember to prepare ~10% more than you think you need. You’ll inevitably spill some when using the Trickle 2 bottle to administer it to the colony. It’s not that expensive, so don’t risk running out.

And the how strong? The recommended concentration to use oxalic acid at in the UK has – for many years – been 3.2% w/v (weight per volume) in 1:1 syrup. This is less concentrated than is recommended in continental Europe (see comments below on Api-Bioxal).

My advice – as it’s the only concentration I’ve used – is to stick to 3.2%.

Listen very carefully, I shall say zis only once

A bit of basic chemistry coming up. Skip to the warning in red below and then the recipes if you want, but this explains some important things about working out how much to use.

The molecular formula of oxalic acid is C2H2O4. The molecular weight of oxalic acid is 90.03 g/mol. However, the oxalic acid you purchase – including Api-Bioxal – is the dihydrated form of oxalic acid.

Di as in two, hydrated as in water.

The molecular formula of oxalic acid dihydrate is C2H2O4.2H2O and oxalic acid dihydrate has a molecular weight of 126.07 g/mol.

Therefore the weight of oxalic acid in 1 g of oxalic acid dihydrate is 90.03/126.07 = 0.714 g.

Caution

Oxalic acid is toxic

  • The lethal dose for humans is reported to be between 15 and 30 g. It causes kidney failure due to precipitation of solid calcium oxalate.
  • Clean up spills of powder or solution immediately.
  • Take care not to inhale the powder.
  • Store in a clearly labelled container out of reach of children.
  • Wear gloves.
  • Do not use containers or utensils you use for food preparation. A carefully rinsed plastic milk bottle, very clearly labelled, is a good way to store the solution prior to use.

Recipes : oxalic acid

The standard recipe is 100 g water plus 100 g white granulated sugar. Mix well and then add 7.5 g of oxalic acid. The final volume will be 167ml i.e. sufficient to treat over 30 seams of bees, or between 3 and 4 strong colonies (including the 10% ‘just in case’).

This final concentration is 3.2% w/v oxalic acid … (7.5 * 0.714)/167 * 100 = 3.2. Check my maths.

0.01 g to 500 g

0.01 g to 500 g

If you have more colonies to treat, or have trouble weighing 7.5g, scale everything up ten-fold. Or buy a small, accurate set of digital scales – like these for £9 which work very well. 1 kg of sugar plus 1 kg (1 litre) of water requires 75 g of oxalic acid and makes 1.67 litres … enough to treat all the colonies in the association apiary.

Which is not such a bad idea. Make it up carefully once and share it with your fellow beekeepers. Storage details are provided below.

Recipes : Api-Bioxal

Warning – the recipe on the side of a packet of Api-Bioxal makes up a much stronger solution (4.4% w/v) of oxalic acid than has historically been used in the UK. Stronger isn’t necessarily better. The recipe provided is 35 g Api-Bioxal to 500 ml of 1:1 syrup. By my calculations this recipe makes sufficient solution at a concentration of 4.4% w/v to treat 11 hives. 

To make a 3.2% Api-Bioxal-based oxalic acid-containing solution using the 35 g pack of Api-Bioxal you need to mix the entire contents of the pack with 691 ml of 1:1 syrup.

Here’s the maths:

  • 35 g of Api-Bioxal contains only 22.14 g of oxalic acid. 88.6% of the 35 g is oxalic acid dihydrate (the remainder is cutting agents like glucose and powdered silica) and so the oxalic acid content is ((35 * 0.886) * 0.714) = 22.14 g.
  • To calculate the volume of syrup you need to divide it by the final percentage required i.e. (22.14 / (3.2/100)) = 691 ml. I don’t know the exact amount of sugar and water needed to make this amount … it’ll be about 430 g of each (I think).

A 35 g packet of Api-Bioxal is therefore sufficient to treat about 15 colonies (assuming 5 ml per seam, 8 seams per hive and 10% ‘just in case’) at the recommended concentration of 3.2% w/v.

Api-Bioxal is sold in three pack sizes (35 g, 175 g and 350 g). If you are wealthy enough to be able to purchase the larger pack sizes you’ve probably got your own beekeeper (or mathematician). Relax on your yacht while they do the calculations for you 😉

On the other hand … if you have a smaller number of colonies either make a full 35 g packet up and share it, or use accurate scales and the following table:

Api-Bioxal recipes for 3.2% OA trickling

Api-Bioxal recipes for 3.2% OA trickling

Storage

Storage of oxalic acid syrup at ambient temperatures rapidly results in the acid-mediated breakdown of sugars (particularly fructose) to generate hydroxymethylfurfural (HMF). As this happens the colour of the oxalic acid-containing solution darkens significantly.

This breakdown happens whether you use oxalic acid or Api-Bioxal.

Stored OA solution and colour change

Stored OA solution and colour change …

HMF is toxic to honey bees at high concentrations. Studies from ~40 years ago showed that HMF concentrations below 30 mg/l were safe, but above 150 mg/l were toxic1. HMF buildup is one way overheated honey is detected.

At 15°C HMF levels in OA solution can reach 150 mg/l in a little over a week. At room temperature this happens much faster, with HMF levels exceeding 150 mg/l in only 2-3 days. In the dark HMF levels build up slightly less quickly … but only slightly 2,3.

Only make up OA solutions when you need them.

If you must store your oxalic acid-containing syrup for any length of time it should be in the fridge (4°C). Under these conditions HMF levels remain well below toxic levels for at least one year. However, don’t store it for this long … use it and discard the excess. Don’t use discoloured oxalic acid solutions as they’ve been stored incorrectly and may well harm your bees.

Please re-read the comments above about the toxicity of oxalic acid. If you are going to store it in the fridge it must be very clearly labelled and there must be no chance that children can reach or open the container.

Conclusions

Api-Bioxal is the least expensive VMD-approved miticide and powdered oxalic acid is much, much cheaper. Both contain the same active ingredient, oxalic acid, which is highly effective against phoretic mites.

In midwinter, with very low levels (or no) of brood, a single oxalic acid-containing treatment minimises mite levels for the coming season.

Oxalic acid-containing solutions are easy to prepare. I recommend you make sufficient for your own colonies and those of your beekeeping friends and association members. My previous BKA used to distribute litres of the stuff for use in midwinter. Use this solution in midwinter and then discard any that is unused.

Oxalic acid-containing solutions are inexpensive and easy to administer by trickling. As I shall demonstrate next time.

Please re-read the safety instructions highlighted in red above.


Michelle Dubois

Michelle Dubois

† Listen very carefully, I shall say zis only once was a catchphrase used by “Michelle of the Resistance” in the 1980’s comedy ‘Allo ‘Allo! Michelle (Dubois) was rarely seen without a trench coat and beret, had a corny French accent and was played by Kirsten Cooke.

‘Allo ‘Allo! ran for 85 episodes in the decade from 1982 on BBC one. It was about a café in Nazi-occupied France and the French Resistance, just about. It mixed bawdy humour with gross stereotypes (posh British twits, sex-obsessed French) and was a parody of ITV’s series Secret Army (’77-’79).

Early episodes had obvious and rather dull titles. In the later series the individual episodes had some quite good puns like Awful Wedded Wife.

Michelle – Listen very carefully, I shall say zis only once

René – Well, in that case, could you please speak slowly?

You had to be there … 😉

‡ Oh alright then, since you insist. The 175 g pack of Api-Bioxal (~£39) needs to be made up in 3.459 litres of 1:1 syrup and the 350 g pack (~£65) 6.919 litres of 1:1 syrup. Determining how much water and sugar to mix to make these amount is, as they say, an exercise for the reader. Assuming a 3.2% solution and 8 seams of bees per colony Api-Bioxal costs between 63p and 41p per hive (see note below), depending upon the pack size you purchase. I know that beekeepers moan on and on about the outrageous cost of Api-Bioxal (as do I), but is 63p per colony really an unreasonable amount to spend on VMD-approved medicines to keep your colony as clear of Varroa as possible? I don’t think so.

Note – the costs in the paragraph were calculated using the lowest prices I could currently find for Api-Bioxal. C Wynne Jones has the 35g packets for £9.50 and Maisemores have the 350g packets for £64.79. Prices correct on 9/10/17.

1 Jachimowich T., El Sherbiny G., Zur Problematik der verwendung von Invertzucker für die Bienenfüttering, Apidologie 6 (1975) 121-143.

2 Bogdanov S., Kilchenman V., Chamere J.D.. Imdorf A. (2001) available online.

3 Prandin, L., Dainese, N. , Girardi, B., Damolin, O., Piro, R., Mutinelli, F. A scientific note on long- term stability of a home-made oxalic acid water sugar solution for controlling varroosis Apidologie, 32:) 451-452

 

Kick ’em when they’re down

Out, damn'd mite ...

Out, damn’d mite …

Why bother treating colonies in midwinter to reduce Varroa infestation? After all, you probably treated them with Apiguard or Apivar (or possibly even Apistan) in late summer or early autumn.

Is there any need to treat again in midwinter?

Yes. To cut a long story short, there are basically two reasons why a midwinter mite treatment almost always makes sense:

  1. Mites will be present. In addition, they’ll be present at a level higher than the minimum level achievable, particularly if you last treated your colonies in late summer, rather than early autumn.
  2. The majority of mites will be phoretic, rather than hiding away in sealed brood. They’re therefore easy to target.

I’ll deal with these in reverse order …

Know your enemy

DWV symptoms

DWV symptoms

The ectoparasite Varroa feeds on honey bee pupae and, while doing so, transmits viruses (in particular DWV) that can completely mess up the development of the adult bee. Varroa cannot replicate anywhere other than on developing pupae. It’s replication cycle, and the resulting mite levels in the colony, are therefore tightly linked to the numbers and availability of hosts … honey bee pupae.

If developing brood is available the mite can replicate. Under these conditions, newly emerged adult, mated, female Varroa spend a few days as phoretic mites, riding around the colony on young bees. They then select a cell with a late-stage larvae in, enter the cell and wait until pupation occurs. If developing worker brood is available each infested cell produces 1 – 2 new mites (drone cells produce 3+) and mite numbers increase very rapidly in the colony.

In contrast, if there’s no developing brood available, the mites have to hang around waiting for brood to become available. They do this as phoretic mites and can remain like this for weeks or months if necessary.

Therefore, when brood is in abundance and the queen in laying freely mites can replicate to very high levels. In contrast, when brood is limiting and the queen has reduced her egg laying to a   v  e  r  y     s  l  o  w     r  a  t  e     the mite cannot replicate and must be predominantly phoretic.

When does this happen?

Lay Lady Lay … or don’t

Ambient temperature, day length and the availability of nectar and pollen likely influence whether the queen lays eggs. When it’s cold, dark and there’s little or no pollen or nectar coming into the hive the queen slows down, or even stops, laying eggs.

About 8 days after she stops laying there will be no more unsealed brood in the colony. About 13 days after that all the sealed brood will have emerged (along with any Varroa). Therefore, after an extended cold period in midwinter, the colony will have the lowest level of sealed brood … and the highest proportion of the mite population will be phoretic.

Under normal (midsummer) circumstances about 10% of the mite population is phoretic. It’s probably unnecessary to state that, if there’s no brood available, 100% of the mites must be phoretic.

All licensed miticides work extremely well against phoretic mites.

Caveats, guesstimates, global warming and the Gulf Stream

Global warming

Global warming …

Whatever the cause, the globe is warming (irrespective of what Donald Trump tweets). Long, hard winters are getting less common (or perhaps even rarer, as they were never particularly common in the UK). In Central, Southern or Eastern Britain it’s possible that the colony will have some brood present all year. In parts of the West, warmed by the Gulf Stream, I’d be surprised if a colony was ever broodless. Only in the North is it likely that there will be a brood break in midwinter.

Most of the paragraph above is semi-informed guesswork. I don’t think anyone has systematically analysed colonies in the winter for the presence of sealed brood. Sure, many (including me) have opened colonies for a quick peek. Others will have peered intently at the Varroa board to search for shredded wax cappings that indicate emerging brood. The presence of brood will vary according to environmental conditions and the genetics of the bees, so it’s not possible to be dogmatic about these things.

However, it’s safe to say that in midwinter, sealed brood – within which the mites can escape decimation by miticides – is at a minimal level.

Reducing mite levels and minimal mite levels

Within reason, the earlier you apply late summer miticides, the better you protect the all-important overwintering bees from the ravages of viruses, particularly deformed wing virus. This is explained in excruciating detail in a previous post, so I won’t repeat the text here.

However, I will re-present the graph that illustrates the modelled (using BEEHAVE) mite levels.

Time of treatment and mite numbers

Time of treatment and mite numbers

The gold arrow (days 240-300 i.e. September and October) indicates when the winter bees are being reared. These are the bees that need to be protected from mites (and their viruses). Mite numbers (starting with just 20 in the hive on day zero) are indicated by the solid coloured lines. The blue, black, red, cyan and green lines indicate modelled mite numbers when the colony is treated with a miticide (95% effective) in mid-July, August, September, October or November respectively.

The earlier you treat, the lower the mite levels are when the winter bees are being reared. Study the blue and black lines.

This is a good thing.

In contrast, by treating very late (the cyan and green lines) the highest mite numbers of the season occur at the same time as the winter bees are being reared. A bad thing.

But … look also at mite numbers after treatment

Look carefully at the mite numbers predicted to remain at the end of the year. Early treatment leaves higher mite levels at the start of the following year.

This is simply because mites escaping the treatment at the end of summer have had an opportunity to reproduce during the late autumn.

This is why the additional midwinter treatment is beneficial … it kills residual mites and gives the colony the best start to the new calendar year§.

Kick ’em when they’re down

Early treatment protects winter bees but risks exposing bees the following season to unnecessarily high mite numbers. However, in midwinter, these residual mites are much more likely to be phoretic due to a lack of brood in the colony. As I stated earlier, phoretic mites are relatively easy to target with miticides.

So, give the mites a hammering in late summer with an appropriate and effective miticide and then give those that remain another dose of the medicine in midwinter.

But not another dose of the same medicine

Since the majority of mites in a colony with little or no brood will be phoretic, you can easily reduce their numbers using a single treatment containing oxalic acid. This can be administered by sublimation (vaporisation) or by trickling (dribbling).

There’s no need to use any treatment that needs to applied for a month. Indeed, many (Apiguard etc.) are not recommended for use in winter because they work far less well on a largely inactive colony.

Trickle 2 - £1

Trickle 2 – £1

I’ve discussed sublimation previously. However, since this requires relatively expensive (£30 – £300) specialised delivery and personal protection equipment it may be inappropriate for the two hive owner. In contrast, trickling requires almost no expensive or special equipment and – reassuringly – has been successfully practised by UK beekeepers for many years. I did it for years before I bought my Sublimox vaporiser.

Therefore, in two further articles this autumn (well before you’ll need to treat your own colonies) I’ll describe the preparation and storage of oxalic acid solutions and its use.

Be prepared

If you want to be prepared you’ll need to beg, borrow or steal the following – sufficient oxalic acid (or Api-Bioxal), a Trickle 2 bottle sold by Thorne’s, a cheap vacuum flask (Tesco £2.50), granulated sugar and a pair of thin disposable gloves.

Do this soon. Don’t leave it until midwinter. You need to be ready to treat as soon as there’s a protracted cold spell (when brood will be at a minimum). Over the last few years my records show that this has been anywhere between the third week in November and the third week in January.

More soon …


† Only MAQS is effective against mites sealed in cells. This is why most miticides are used for extended periods in the late summer or early autumn … the miticide must be present as Varroa emerge from sealed cells.

‡ I’ll repeat the caveat that this is an in silico simulation of what happens in a beehive. Undoubtedly it’s not perfect, but it serves to illustrate the point well. It’s freely available, runs on PC and Mac computers, and is reasonably well-documented. In the simulations shown here the virtual colony was ‘primed’ with 20 mites at the beginning of the year. BEEHAVE was run using all the default settings – climate, forage etc. – with the additional application of a miticide (95% effective) in the middle of the months indicated. Full details of the modelling have already been posted.

§ The National Bee Unit recommend Varroa levels are maintained below 1000 throughout the season. Without treatment, 20 mites at the start of the season can easily replicate to ~750 in the autumn. If you start the season with 200 mites then levels are predicted to reach ~5000 in the following summer. The colony will almost certainly die that season or the next. There’s a more detailed account of the consequence of winter brood rearing and the level of mite infestation written by Eric McArthur and reproduced on the Montgomeryshire BKA website that’s worth reading.

¶ The cumulative (year upon year) effect of late summer treatment with no midwinter treatment has been discussed previously. I’ll simply re-post the relevant figure here – 5 years of bee (in blue, left axis) and mite (in red, right axis) numbers with only one treatment per season applied in late September. Within two years the higher mite numbers that are present at the start of the year reproduce to dangerously high levels.

Mid September

Mid September

Counting Varroa

It’s that time of the season again. With the exception of readers in the Southern Hemisphere, Colonsay, the Isle of Man or a few favoured locations in the Highlands of Scotland, miticide treatments should be on to reduce Varroa levels.

For reasons explained elsewhere, it’s important that this is done before the winter bees are exposed to the smorgasbord of viruses that Varroa transmits when it feeds.

It’s not sufficient to just treat. You also need to have some idea that the treatment is reducing the numbers of Varroa in the colony.

Counting by numbers

It has been determined that only 10-20% of mites in a colony are phoretic i.e. attached to emerged workers. The majority of treatments (MAQS is the current exception) only target these mites. Therefore, treatments are usually applied over a period of several weeks to ensure that mites newly emerged from capped cells are also exposed.

There are a couple of obvious ways to determine the mite load before and after treatment. These include:

  • conducting an alcohol wash test, or a sugar-roll equivalent, of workers to quantify the phoretic mites.
  • uncap a known amount of worker brood (drone brood is almost certainly absent from colonies this late in the season) to quantify mite infestation.

However, both are pretty intrusive and – with the exception of the sugar-roll – involve the sacrifice of bees or brood, so perhaps not ideal at this stage of the season. However these are the most accurate way of measuring things.

Counting the corpses

Out, damn'd mite ...

Out, damn’d mite …

Alternatively, and this is what most beekeepers do, apply the treatment and count the mite drop.

To count the mites you need some way of catching the mites. Open mesh floors (OMF) can easily be fitted with a sheet of closely-fitting (most usefully white) Correx onto which the mites drop. Restrict the access of ants and other creepy crawlies to the tray or they may steal some of the corpses. Check these on a regular basis during treatment and you have a simple way of determining whether the treatment is working.

The treatment may be working, but has it been effective?

The scores are on the floors

If you count thousands of dropped mites and that number doesn’t diminish during treatment i.e. the drop per day early and late in treatment is broadly similar, then the treatment is working, but it’s not effective or finished as there are loads of mites still left.

What you need to observe is a reduction in mite drop when comparing early and late counts.

Depending upon the treatment, the first days’ drop isn’t necessarily indicative of whether the miticide is working (or of the phoretic mite load of the colony). It may take a day or two for the treatment to achieve maximum kill. Vaporised oxalic acid often gives a better drop after 24-48 hours, and continues to work over about 5 days.

As indicated in the footnote, the numbers of brood emerging per day will expose ‘new’ mites to the miticide, increasing the count. If emerging brood levels vary, so will the mite drop … but also remember that the efficacy of the miticide also varies over time.

What you’re looking for is a hugely reduced count of mites dropped per day at the end of the full treatment period when compared with the beginning.

I usually carefully monitor the first week or two and the last week. Simples.

Objective vs. subjective counting

Easy counting ...

Easy counting …

Some beekeepers count each and every mite that appears on the trays. Others just look for ‘lots’ at the beginning and ‘almost none’ at the end. I consider >50/day is ‘lots’ and only count smaller numbers.

The less frequently you count the more difficult it is to discriminate dead mites from all the other detritus that accumulates on the trays. The cell cappings, the pollen that’s being dropped, the wax scales and various other bits of bee, all make spotting the mites more tricky.

The larger the area you’re counting the more likely it is to either double-count or miss mites. Make life a bit easier by ruling a simple grid onto the tray and counting square by square.

Scrape the tray clean after counting the mites … if you leave the tray dirty you’ll end up double counting and struggling to spot mites that are knee-deep in the crud that’s fallen through the OMF.

Don’t try this at home

Varroa are a pretty regular size and shape. And colour for that matter. At least adult mites are. This raises the possibility – though perhaps only to those with a tendency towards geekiness – to try and count mites automagically§.

Rather than stand around the apiary squinting through myopic eyes at tiny reddish ovals you could simply photograph the tray and then process the image later.

Been there, done that … or at least tried to.

Fiji ...

Fiji …

There’s a freely-available, well-supported, image analysis package called ImageJ (also distributed sometimes as the auto-referential Fiji … Fiji is just ImageJ). It’s possible to count objects using ImageJ having set criteria that define them.

As an exercise in near-futility I’ve attempted to do this for Varroa. You first need to ensure the Varroa are of a standardised size and shade by scaling the image appropriately and correcting the colour. This can be done by using a photographers grey card of a known size, placed to the side of the Varroa tray. You then use this as a reference to scale the image and define the white balance.

Finally, you define the size, roundness and shade of a Varroa and process the image in ImageJß. It counts the mites and provides an overlay with each identified mite numbered. You’re then able to check whether it’s missed any.

It does.

Consistently variable

This is the point I’ve got stuck at … the accuracy is all over the place but it’s clearly not impossible. Problems include:

  • It overlooks mites lying on their ‘edges’, perhaps propped up on a speck of pollen or fragment of wax. Better colour definition and a wider range of ‘ovality’ might sort this out.
  • It misses mites lying immediately next to another mite – these look like 8 or ∞ rather than a simple solid oval. I’ve no clear solution to this other than counting lower densities of mites.
  • It ignores some mites that appear as ‘doughnuts’ because of reflection from the shiny carapace. Don’t use flash for the photography.
  • It counts some ovalish, reddish lumps of pollen that are about the right size as mites. D’oh!

At best the accuracy is above 80%, but it’s variable. The lack of consistency is the major issue. If it was always 80% it would be perfectly acceptable and a very fast way to record mite numbers. At worst – usually when the tray is messy and mite numbers are relatively low – it’s well below 50%.

This is an intriguing beekeeping-related task for long winter nights. If you’re a geek. My ambition is to take a quick smartphone photo, scrape the Correx tray clean and then (automagically!) do the counting at home with a cup of tea and piece of cake.

I’ll keep persevering … particularly with the tea and cake 😉


† It’s currently Spring in the Souther Hemisphere, so the wrong time to treat. The remaining locations (and Australia) have no Varroa so have no need to treat. Lucky blighters.

‡ This is a gross oversimplification. Obviously, a broodless colony will only have phoretic mites. Swarms that issue from colonies take 35% of the mites with them, leaving 65% on the remaining bees (or capped in cells). The actual number of phoretic mites likely depends upon the prior history of egg laying by the queen. It also is probably influenced by the overall level of mites in the colony (or ratio of uncapped brood to mites perhaps). I’m not sure if anyone has modelled this successfully, though it might be possible to do this with BEEHAVE.

§ Automagically is pretty obviously a concatenation of automatic and magic. It is usually defined as “(especially in relation to the operation of a computer process) automatically and in a way that seems ingenious, inexplicable, or magical”. Interestingly, the term was first used in the 1940’s, well before the advent of computers.

ß Once I’ve got this working better I’ll provide some instructions … in the meantime the menus that you need to use are Analyse … Set Measurement and Analyse … Count Particles. Image scaling needs to be done first in ImageJ. Currently I do the white balance in Adobe Lightroom (which is overkill, but convenient as all my images go through this software).

 

Right here, right now

In February 2016 I posted an article on When to treat colonies with miticides. It was read by subscribers, generated a bit of discussion – in particular on Apiguard use – and then disappeared into the howling wilderness that is the interwebs …

Google hides the searches that drives most of the website traffic to this site. However, it has been found … as is clear from the access stats (below), it is now being accessed extensively.

This reflects the change in the seasons as beekeepers turn their thoughts from harvesting honey to protecting their colonies from the ravages of Varroa and the viruses it transmits.

'When to treat' stats ...

‘When to treat’ stats …

It’s worth reiterating it here, though this won’t be news to regular readers, Varroa itself is probably not the problem. The problem is the smorgasbord of viruses that Varroa transmits when feeding on the haemolymph (blood) of honey bee pupae.

Benign and virulent

Most important of these viruses is deformed wing virus (DWV). This virus has a sort of Jekyll and Hyde personality. It’s probably present in all honey bees, transmitted between bees while larvae are being reared and during trophylaxis (the regurgitation of liquid food between bees). Under these conditions, and in the absence of Varroa the virus is probably benign.

DWV symptoms

DWV symptoms

Of course, it’s difficult to test whether it is really benign as there probably aren’t any bees that lack DWV. Even bees that have never been exposed to Varroa, such as the black bees on Colonsay, have DWV. Let’s assume that, even if not benign, it has minimal detrimental effect on the bees.

Varroa changes the route by which DWV is transmitted. Instead of being orally transferred – a route the bees have probably evolved to cope with – Varroa bypasses any defence mechanisms by ‘injecting’ DWV directly into the blood. Under these conditions DWV reproduces rampantly – for reasons that have yet to be determined – causing the ‘deformed wing’ symptoms most beekeepers are familiar with.

Symptomatic adult, or recently emerged, bees can contribute little or nothing to the colony.

Live fast, die young

But there’s more bad news. Asymptomatic adult bees with high levels of DWV have a shorter lifespan and die prematurely. This is perhaps not an issue during the heady days of summer when the turnover of worker bees is at its height – the queen is laying well, perhaps 1500-2000 eggs per day, the colony is bulging and individual workers “live fast and die young” after about 6 weeks.

Formally, I don’t think it’s been shown that mid-season workers have a shorter life span when they have high levels of DWV. What is known, and what is much more important, is that winter bees die prematurely if their DWV levels are high.

Winter bees are the ones with high levels of fat in their bodies. These are the bees that get the colony through the winter. Some might live for 5-6 months in the UK, and it’s known they can live for up to 9 months. If these bees die early, in the absence of any significant brood rearing, the colony dwindles and dies.

Game over.

Preventing the inevitable

Varroa transmits DWV and results in high levels of DWV. High levels of DWV in winter bees shortens their lifespan and results in colony losses. How can you prevent the inevitable?

In the absence of ways to directly control DWV levels (these are in development but you’re then tackling the symptom, not the cause) the only way to do this is to prevent the transmission of DWV to the winter bees by Varroa in the first place.

And you do this by applying effective miticides early enough that the winter bees are protected from exposure to Varroa, and the viruses it transmits.

How early is early?

I discussed this in the earlier article and am working on a more nuanced version at the moment. Essentially – and I’m writing this in mid-August – the answer is now or very soon.

In the definitive publication demonstrating the premature death of winter bees by DWV, Peter Neumann and colleagues detected a measurable reduction in longevity as early as November in the colonies they studied. These bees were age-marked and had emerged 50 days earlier. The eggs had therefore been laid in the first week of September … and been capped (together with any Varroa) as pupae in mid-September.

Therefore, treatments to reduce Varroa should be completed by mid-September to protect the winter bees. Since many treatments take ~4 weeks the time to treat is right now.

Caveats

There’s climatic variation between parts of the UK and Bern, where the study by Peter Neumann was conducted. There’s also seasonal variation year on year.

In the balmy South the dates will be later than in the frigid North. In cool years with an early autumn they will be earlier, whereas an Indian summer will delay the need to treat.

Treatments are generally incompatible with a honey flow. If you take your bees to the heather you have to balance collecting a late heather crop with protecting the bees from the ravages of Varroa.

it’s not possible, or wise, to be dogmatic about precise dates … other than to say that miticide treatment is generally required earlier than you think to protect the winter bees from DWV.

Better late than never?

Well, yes, but the damage may well have been done.

The annual survey of beekeepers in Scotland regularly includes significant numbers using Apiguard in October. Close, but no cigar (actually, not even close) … it’s probably too late to reduce Varroa levels in a meaningful way, and it’s unlikely to be effective anyway as Apiguard needs average temperatures around 15°C.

All of the comments above on the timing of treatment make the assumption that the treatment is effective – the right dose, the right duration, the absence of resistance etc.

Finally, it’s worth noting that starting treatment in mid/late August does not reduce Varroa levels to the lowest achievable levels. Treating later in the year does this, because more mites are phoretic and ‘reachable’ by the treatment. To reduce mite levels to the minimum you also have to also treat midwinter … something for another post.


† Live fast, die young was the title of a biography of the actor James Dean by James Gilmore. It’s a popular phrase, being used for a movie and several song titles. The extended version Live fast, die young and have a good looking corpse, often wrongly attributed to James Dean, actually came from the 1947 book Knock on Any Door by Willard Motley.

‡ Close but no cigar is a mid-20 phrase from the USA. It dates back to the time when fairground stalls gave out cigars as prizes.

Colophon

Right here, right now is a song released in April ’99 by Fatboy Slim (Norman Cook) from the album You’ve come a long way, baby‘. If you appreciate evolution you’ll enjoy the video …

… but you’ll need to like beat/dance music to appreciate the track.

Size matters

Anyone reading the beekeepingforum.co.uk will be aware that there are a number of contributors there that enthusiastically recommend the treatment of colonies with vaporised (or, perhaps more accurately, sublimated) oxalic acid to reduce Varroa levels.

There goes a few pence ...

There goes a few pence …

Although vaporised oxalic acid (OA) has been used by some for many years, the speed with which it has recently been embraced by many UK beekeepers (at least those that contribute to discussion forums and, perhaps to a lesser extent, those I speak to in associations over the winter) probably reflects two or three things:

  • an awareness of just how effective oxalic acid is as a treatment
  • the increased availability of commercial oxalic acid vaporisers (or Heath Robinson-like plans to build-your-own)
  • the huge price-differential between oxalic acid and most other treatments

There are almost as many homegrown or imported vaporisers as there are treatment regimes to hammer down the mite levels. Of course, there’s the contentious point that oxalic acid is not approved by the VMD (Veterinary Medicines Directorate), despite having been in routine use for decades. Api-Bioxal is, but is probably unsuitable for sublimation due to the inert (as far as Varroa are concerned) additives it contains. Api-Bioxal can be vaporised but leaves a caramelised residue in the vaporiser pan that is hard to clean.

Out, damn'd mite ...

Out, damn’d mite …

‘Vaping’ is also popular in the US. Randy Oliver has covered it extensively on his scientificbeekeeping.com site and it’s also regularly discussed on Beesource. OxaVap make/supply a vaporiser that appears very similar to the Sublimox I use. The OxaVap model has a useful temperature display that I would find much easier to read than the red/green diodes on the Sublimox … I’m red/green colourblind.

Active and passive vaporisers

The Sublimox and OxaVap vaporisers are ‘active’ … they blow out a dense cloud of OA-containing vapour through a relatively narrow diameter nozzle (the video below uses water to demonstrate this process). This provides advantages both in terms of ease and speed of delivery. These vaporisers simply need a 7mm hole drilled through the sidewall of the floor (see photo at the top of the page), or through an eke placed over the colony. The OA-containing vapour is ‘squirted’ in, permeates all corners of the hive within seconds and you can then move on to the next hive. The vaporiser doesn’t need cooling between treatments and the dose administered is tightly controlled.

Big Daddy

However, OA dosage isn’t critical. It has been shown to be well-tolerated by bees in studies from groups in the UK and Germany. If the dose isn’t critical and speed really is important then perhaps consider the vmVaporizer. At $3600 it’s about ten times the price of a Sublimox.

vmVaporizer ...

vmVaporizer …

The manufacturers claim you can treat 300 hives an hour with one of these … one every 12 seconds. For comparison, the Sublimox takes 20-30 seconds per hive. However, what takes the time is sealing the hive, moving the generator about, unsealing the hive etc. so you’d need a team of (well protected) helpers and some closely spaced hives to achieve a similar rate. The vmVaporizer is mains (110V) powered so would also need a generator or inverter.

The video above demonstrates the vmVaporizer in action. It produces copious amounts of oxalic acid vapour, albeit less ‘forcefully’ than the Sublimox. It seems the only way to control how much is delivered is by changing the duration the hive is exposed for.

Undoubtedly this is overkill for the majority of readers of this site, but it’s interesting to see what the commercial beekeeping community are using (much like browsing the decapping or bottling machines in the Swienty catalogue). There’s at least one satisfied UK-based beekeeper quoted on the vmVaporizer site so … Mark, if you happen to read this I’d be interested in how well the machine works and whether you can achieve the quoted hive treatment every 12 seconds?

And, does it work with Api-Bioxal?

😉

 

Apistan redux†

I’ve discussed Apistan, a pyrethroid treatment for Varroa, in two recent posts. In these I explained in some detail its molecular mechanism of action. I also explained the two major problems associated with Apistan (and the related tau-fluvalinates ) – the widespread resistance of Varroa to Apistan and the residues it leaves in wax.

In this final post I’m going to revisit just how useful Apistan could be if it was used in a more rational manner. I’m going to concentrate on resistance and you’ll probably need to read the previous post on this topic to provide necessary the background. I’ll only really touch on the residues in wax at the end – I’ve already discussed how these can be minimised if you consider them an issue.

This is (another) long post. It draws together the concepts described in previous articles and links the science of Varroa control to potential strategies to benefit practical beekeeping.

How good is Apistan if Varroa are not resistant?

Apistan

Apistan

Exceptionally good. Pyrethroids are some of the most widely used pesticides. They are widely used because they are very effective. Apistan is no exception. When used to treat Varroa populations that are not already resistant it kills over 98% of the mites in the colony when used according to the manufacturers instructions. 98% … that reduces the National Bee Units’ recommended maximum mite load of 1000 to just 20.

Just how effective is emphasised by a quote from the Apidologie paper cited above. “In treated hives, worker pupae and adult bee infestations decreased from 14.2 ± 7.3% to zero and from 15.7 ± 7.3% to zero, respectively. Whereas, in the two control hives, during the first 6 weeks, the average worker pupae infestation increased from 15.9 ± 2.9% to 19.7 ± 3.5%”.

Most mite mortality occurred during the first 4 weeks of treatment and the level of Apistan present at the beginning and end of treatment remained at about 10% i.e. it should be as active at the end of the treatment period as at the beginning.

How good is Apistan in reality

Resistance was first demonstrated in 2002 and is now widespread in the UK. In a recent paper, Ratneiks and colleagues (University of Sussex) demonstrated that Apistan was significantly less effective at killing Varroa when used for a second treatment, four months after the first. In this study they showed only 33% of mites were killed at the second treatment, whereas 58% were killed in colonies treated for the ‘first time in five years’.

This isn’t rocket science … if there are some resistant mites in a population then Apistan will preferentially allow these to survive. Consequently they will make up a greater proportion of the mite population when re-treated.

Since we know the molecular basis of resistance to Apistan it would now be possible to determine – without doing the treatment and counting the corpses – what proportion of mites were resistant in a population before treatment. It would therefore be easy to determine whether treatment would be likely to work.

Equally, it would be possible to determine whether the colonies ‘not treated with Apistan for five years’ still maintained significant levels of Apistan resistant mites. As will become clear, there are studies that contradict this, and the definitive test – the presence of absence of the mutation that confers resistance – was not done in the Sussex study.

Apistan resistance and fitness costs

Mutations, such as the one that confers resistance to Apistan, can – in broad terms – exert three different effects:

  1. Beneficial – the presence of the mutation favours the organism (a fitness benefit), the mutation will be selected for and it’s presence in the population is likely to increase.
  2. Detrimental – the mutations causes a fitness cost and organisms that carry it are likely to reproduce less well, resulting in it being lost from the population.
  3. Neutral – the mutation is neither beneficial nor detrimental.

In the presence of Apistan, the Leucine to Valine mutation at residue 925 (L925V) of the voltage gated sodium channel (VGSC; please see the previous article on the molecular basis of resistance), is a beneficial mutation. Any mites that carry it will not be killed and will be able to reproduce, so increasing it’s prevalence in the population. The same reasoning applies to other Apistan resistance mutations.

The VGSC of Varroa evolved over eons in the absence of Apistan. The mutation is in a part of the protein critical for its function (that’s why Apistan binding blocks function). It’s therefore perhaps unsurprising that in the absence of Apistan selection there is evidence that the L925V mutation is detrimental. In simple terms the VGSC works less well with a Valine at position 925 than a Leucine unless Apistan is present. Where’s the data that supports this?

The influence of prior treatment on Varroa genotype

Table 1. Apistan resistance mutations in Varroa from treated and untreated colonies

Table 1. Apistan resistance mutations in Varroa from treated and untreated colonies

The table above needs a little explanation. Colonies from Henlow and Shillington were treated with Apistan and tested one month later. Colonies from Harpenden, Bishop Stortford, St. Albans and Peterborough had no history of Apistan treatment in the recent past. Unfortunately, the paper does not make clear when the last treatment was, with the exception of a sample from Harpenden which had not been treated for at least 3 years.

Varroa is diploid i.e. there are two copies of the gene for the VGSC. The S and R heading the columns SS, SR, RR, indicates whether the Apistan resistant mutation is absent (S = sensitive) or present (R=present). SR indicates that the mite was heterozygous, one resistant copy and one sensitive. Whether these mites have lower resistance than RR mites has not been determined – for the purpose of the remaining discussion I’m going to lump the SR mites with the RR mites and assume they are resistant§.

Of 279 mites tested, 40 were from Apistan-treated and 329 from -untreated colonies. Of the 40 mites from Apistan-treated colonies, all contained the mutation conferring resistance to the fluvalinate. Of the 239 mites from colonies not recently treated with Apistan, 215 were sensitive and only 25 were resistant.

This suggests that in the absence of Apistan, Varroa sensitive to the fluvalinate replicate better.

Is this a surprise?

No. Partly for the reasons explained above … the Leucine at position 925 is likely to stop the VGSC working as well. More compellingly though is the wealth of data suggesting that insecticide resistance is associated with fitness costs in a range of other insects.

Colorado beetle

Colorado beetle

For example, pyrethroid resistant Myzus persicae (peach-potato aphid) exhibit fitness effects in overwintering survival, response to aphid alarm pheromone and vulnerability to parasitoids; pyrethroid-resistant Cydia pomonella (codling moth) have reduced fecundity, body mass of instars, adult male longevity and larval development; finally, pyrethroid-resitant mutants of the snappily-named Leptinotarsa decemlineata (which you of course know as the stripy-attired Colorado beetle) have reduced fertility and fecundity.

Google will find relevant reference on all the above examples or you can refer to a concise mini-review by Kliot and Ghanim Fitness costs associated with insecticide resistance published in Pest Management Science (2012) 68:1431-37.

Before discussing implications for practical beekeeping I should add that the rate at which the loss of the L925V mutation, and other mutations associated with Apistan resistance, needs to be accurately determined. If, as looks likely, a period of 3+ years results in selection for the sensitive variant of the VGSC, it might be possible to develop rational Varroa treatments that exploit this.

Apistan resistance, rational Varroa control and practical beekeeping

For the sake of discussion, let’s accept the following statement:

  • Apistan is devastatingly effective on sensitive mite populations.
  • Apistan is much less effective (or almost completely useless) on resistant mite populations.
  • Resistance by Varroa is acquired rapidly and lost over the subsequent 2-3 years in the absence of selection.

An effective and rational Varroa control strategy would only use Apistan once every 3-4 years, alternating it with other treatments. To mitigate the transfer of Apistan-resistant mites between colonies due to drifting and robbing, or due to the movement, sale and/or relocation of hives during the season, Apistan use would have to be coordinated. This coordination would have to be both geographical and temporal. There would be no point in the Fife beekeepers using it one year if the Angus beekeepers planned to use it the following year.

“Like herding cats” I hear some mutter …

Perhaps, but the benefits would be considerable. How could it be achieved? Perhaps by restricting the sale of Apistan to certain years, in a formulation or package that meant it had to be used quickly or became inactive.

What about the residues in wax?

I’m not sure whether the level Apistan accumulates to in wax is sufficient to be a selective pressure on the mite population. Apistan strips are 10% Apistan. Nothing like that much accumulates in wax. In a recent study fluvalinate levels ranged between 2 and 200,000 parts per billion in wax (mean ~7500 ppb). However, it is a valid concern and so would necessitate a relatively simple experiment to determine the rate at which Apistan resistant mutations are lost in the presence of absence of trace levels of Apistan in comb.

Herd immunity and the responsibility of the individual

There’s a debate in human healthcare about the necessity to vaccinate individuals in a well-vaccinated population. The chance of an infectious disease spreading to the unvaccinated individual in a protected population is very slight. So, why vaccinate?

Well, what if increasing numbers decided not to vaccinate? Once protection in the population falls below a certain level there is a significant chance that an infectious disease will spread widely. We saw this in the UK after the MMR (measles, mumps and rubella) vaccine was falsely claimed to be associated with autism. Vaccination rates dropped from 90+ percent, to low 80’s and – in parts of the country – to only 60%. Unsurprisingly, measles cases increased and – tragically, for the first time in years – there were childhood deaths due to measles infection.

This may seem a million miles away from looking after our bees, but there are parallels. As beekeepers we have responsibility for our own stock. We also have responsibility to the wider community of beekeepers which – because of the way our bees forage and mingle – happily exchange pests and pathogens.

Beekeepers who do not control Varroa (and consequently virus) levels threaten the viability of their own colonies and those of other beekeepers in the area. The same applies to the foulbroods. This is why the bee inspectors try and check all colonies in the vicinity of an outbreak. This is why standstill orders are placed on apiaries where outbreaks occur.

Perhaps this sort of communal responsibility also applies to Varroa treatment using Apistan? Beekeepers who treat without demonstrating very high levels of susceptibility first in their stocks are simply selecting for resistant mites, reducing the efficacy of treatment for themselves, and others, in the future. Indiscriminate or incorrect use of Apistan has resulted in widespread resistance, thereby compromising Varroa control for all beekeepers.

The coordination and control, geographically and temporally, of Apistan usage would benefit beekeeping and beekeepers.

And … it would also benefit those who chose never to treat with Apistan. Treated colonies in the one year in three Apistan was used would have very low mite levels. Fewer mites would be transferred from these colonies by drifting or robbing … what’s not to like?


 Redux, as in the literary term meaning brought back or restored, derived from the Latin reducere (to bring back).

 This is one compelling reason why Apistan strips should not be left in the colony longer than is recommended. It kills the susceptible mites within the first month or so. After that it effectively selects for resistant mites, allowing them to replicate.

 With apologies to any population biologists who were reading this and have now given up in horror.

§ And I’ll save discussion of the influence of the incestuous lifestyle of Varroa and Varroa levels on the ratio of homozygotes to heterozygotes at different stages of the season for a later post. It’s a fascinating and at the same time rather sordid tale …

 Or 4 or 5 – this would need to be determined empirically.

Apistan resistance

Apistan

Apistan

In an earlier article I discussed what Apistan is – a pyrethroid miticide – and the consequences of using it. These include decimation of the mite population if it is susceptible, coupled with the accumulation of long lasting residues in wax. These residues may adversely effect queen and drone development. I also discussed ways to avoid build-up of Apistan residues in comb.

The key phrase in the paragraph above is ‘if it is susceptible’. Unfortunately, resistance to Apistan and the related tau-fluvalinates develops very quickly. To understand why we’ll need to look in a little more detail at how Apistan and other pyrethroids work.

How does Apistan work?

Apistan, like other pyrethroids, works by blocking the activity of voltage gated sodium channels (VGSC) resulting in paralysis because the axonal membrane cannot repolarise.

What on earth does that mean?

Action potential

Action potential

Nerve transmissions – like the signal from the Varroa brain to tell the Varroa legs to move – travel along axons. These are usually very long thin cells. In the adjacent image the ‘brain’ is on the left and the leg muscles on the ‘right’. The nerve impulse (the moving arrow) travels down the axon ‘driven’ by a change in polarity (charge) across the membrane of the axon. In the resting state, when there is no impulse, this is positively charged on the outside and negatively charged on the inside. Sodium – remember the ‘S’ in the acronym VGSC – is positively charged and crosses the membrane (out to in) via a small pore or hole as the impulse passes. This makes the inside of the axon transiently positive. The pore or hole is the VGSC.

Top view of a VGSC

Top view of a VGSC

The VGSC is a transmembrane protein. It actually crosses the membrane multiple times and assembles to form a very narrow channel through which the sodium passes. The cartoon on the right shows the top view of a VGSC, looking “down” the pore into the inside of the axon. The blue bits can move to open or close the pore, allowing sodium to traverse – or not – the membrane into the axon. Apistan binds to the transmembrane protein and prevents the pore from closing. As a consequence, sodium continues to pass from the outside to the inside of the axon, the nerve cannot repolarise and no further impulses can be transmitted. As a consequence, Apistan paralyses the Varroa.

But I don’t suppose many beekeepers will feel much sympathy for the mite 😉

Why isn’t the beekeeper paralysed as well?

Nerve impulses in Varroa and humans are transmitted in essentially the same way. We also have VGSC’s that operate in a similar manner. Why doesn’t Apistan also paralyse careless beekeepers? More generally, why are pyrethroids the most widely used insecticides, available in all garden centres and supermarkets?

Two factors are at work here. The first is the specificity of binding. The VGSC is a protein. Proteins are made from building blocks termed amino acids. The precise sequence, or order, of amino acids is usually critical for protein function. However, two proteins with a similar function can exhibit differences in the amino acid sequence. Although the human and mite VGSC have a similar function they have a different amino acid sequence. Apistan binds much better to the mite VGSC than the human VGSC (this also explains why bees aren’t also paralysed by Apistan … the miticide is specific for the mite VGSC and binds poorly to the honey bee VGSC). In addition, many mammalian species have a number of detoxifying enzymes which deactivate pyrethroids, rendering them ineffective. Together, this explains the specificity of Apistan and other pyrethroids, and the low level of toxicity to humans.

So now you know how Apistan works we can address the much more important question …

Does Apistan work?

Unfortunately, usually not. Since the late-1990’s there have been a large number of publications of Apistan- or fluvalinate-resistant mites from many countries, including the USA (1998, 2002), Israel (2000), UK (2002), Spain (2006), Korea (2009) and Poland (2012). The National Bee Unit used to report Varroa resistance test results by geographic region in England and Wales. Resistance was first reported in mites from Cornwall and Devon (in 2001 and 2002). By 2006 resistance was very widely distributed throughout England. By then approximately a third of all mite samples tested were resistant. The number of tests conducted (or at least reported) then dwindled and there have been none reported since 2010. Not no resistance … no tests. Presumably it’s no longer worth reporting as resistance is so widespread.

The most up-to-date map on the distribution of Apistan resistance I could find is in the NBU booklet on Managing Varroa [PDF; page 28 of the 2015 edition], though the data presented is from 2009.

However, bee equipment suppliers continue to sell Apistan (even Vita, the manufacturer, states that resistance is widespread) and beekeepers continue to use it. Many do so without first testing whether the mite population in their colonies is sensitive to the miticide. How should this be done?

Testing for resistance

Vita suggest two tests. Their first (the “rule of thumb test”) is deeply flawed in my view. It suggests simply looking for a drop of 100’s of mites in the first 24 hours after treatment starts as indicative of a sensitive population.

This isn’t good enough. What if there were thousands of mites present? Perhaps 20% of the population are sensitive, with the remainder resistant. 20% of 5000 mites is 1000 … so you might expect a drop of 100-200 (the majority of the phoretic population) within the first 24 hours. Some might consider this drop indicates a sensitive population … it doesn’t.

It’s not sufficient to count the corpses … you need to know how many mites were unaffected by the treatment.

The second Vita-recommended test is a cut-down version of the “Beltsville” pyrethroid resistance test which is fully described in an NBU pamphlet (PDF). This is much more thorough. Essentially this treats ~300 bees with Apistan, counts the mites that are killed in 24 hours and then counts the unaffected mites remaining on the bees. It’s only by knowing the total number of mites at the start and by determining the percentage of mites sensitive that you can be sure that the treatment is effective.

What is the molecular basis of resistance?

We’re almost there … specific pyrethroids, like Apistan, bind to specific parts of the VGSC. The VGSC is a protein made up of a long connecting chain of amino acids. The binding of the pyrethroid requires an interaction with a small number of specific amino acids in the VGSC. If these particular amino acids change – through mutation for example – then the pyrethroid will no longer bind. If the pyrethroid does not bind the VGSC can open and close again, so the axon repolarises and the mite is not paralysed. The mite is resistant and can then go on to rear lots more resistant baby mites … which, in due course, transfer the viruses that kill your bees.

And that’s exactly what happens.

Leucine

Leucine

A single mutation that causes a substitution of amino acid number 925 in the Varroa VGSC, which is usually a leucine, to either a valine, a methionine or an isoleucine, is sufficient to prevent Apistan and other tau-fluvalinates from binding. At least 98% of mites resistant to Apistan have one of these substitutions. Apistan resistant mites with substitutions at position 925 have been found in the UK, eastern Europe and several sites in South-Eastern USA. It wouldn’t be surprising if the remaining ~2% of resistant mites had a mutation at one of the other amino acids involved in pyrethroid binding. Further studies will confirm this (there are alternative mechanisms that cause resistance, but the one described here is the most frequently seen).

Why aren’t all Varroa mites resistant to tau-fluvalinates?

Apistan resistance has clearly been demonstrated for the last two decades. Resistance is easy to acquire and selection – in the presence of the pyrethroid – is effectively absolute. Without the necessary mutation the mites die, with the mutation they survive.

Bees – and the phoretic mites that are associated with them – are moved around the place all the time, by migratory beekeepers, by importers and through robbing and drifting between colonies.

Why therefore aren’t all Varroa mites now resistant to Apistan and other tau-fluvalinates?

The answer to that is interesting and suggests strategies that could make Apistan an effective treatment again … but I’ll save that for another time.


Only transiently as the charge is reversed shortly afterwards by a similar, though not identical,  mechanism that does not use the VGSC. However, life is simply too short to describe this bit as it’s not needed to understand pyrethroid – or Apistan – activity and resistance.

 The incestuous life cycle of the Varroa mite is important here. This post is already too long to fully elaborate on this but the size of the mite population relative to available open brood (and whether you get single or multiple occupancy of cells) will likely influence the proportion of resistant, partially resistant and sensitive mites in a population.

Credits – the action potential GIF was created by Laurentaylorj from Wikipedia.