Category Archives: Science

Hanging around

I’ve recently discussed the misnamed ‘phoretic’ phase in the life cycle of Varroa destructor. Here we’ll briefly explore some features of this important, but non-reproductive, phase. It’s important because, as I’ll show, it influences subsequent mite reproduction.

I won’t rehash the life cycle of Varroa in detail as I’ve covered it previouslyVarroa is an ectoparasite of honey bees. It reproduces in capped cells, feeding on the developing pupa. A mated female mite enters the cell a few hours before capping and she and her incestuously mated daughters are released when the bee emerges.

The longer pupal development takes, the more progeny mites are produced, so Varroa has evolved to preferentially infest drone brood.

A smorgasbord of viruses

As an ectoparasite of honey bees, Varroa is responsible for the transmission of a smorgasbord of pathogenic viruses to the developing pupa. Subsequent virus replication, particularly by the aptly-named deformed wing virus, can result in developmental deformities.

Worker bee with DWV symptoms

Worker bee with DWV symptoms

Emerging workers with deformities are rapidly ejected from the hive. Other infested workers, with high viral levels, have reduced longevity. This is probably what accounts for the majority of overwintering colony losses. It may also explain so-called ‘isolation starvation‘.

The ‘phoretic’ phase

Mites outside capped cells are termed ‘phoretic’ mites. Recent studies have indicated that these mites are feeding on the workers to which they are attached. The same studies have shaken the long-held assumption that Varroa feeds on haemolymph 1 by rather neatly demonstrating that it is the fat body tissue of the bee that is the plat du jour.

The duration of the ‘phoretic’ phase is dependent upon the state of the colony. Since it is defined as the phase in which mites are not associated with developing pupae, in a broodless colony all the mites are ‘phoretic’. Under these circumstances mites remain ‘phoretic’ until either brood is produced, or they fall (or are groomed) off and drop through the open mesh floor.

The duration of the ‘phoretic’ phase

In colonies with ample brood the ‘phoretic’ phase is, on average, 6 days in length. The range often quoted is 4 – 11 days. The absolute figure must depend upon a number of factors. These include the chance of a mite encountering a late-stage larva. This is presumably influenced by the amount of suitable-aged brood in the colony and – because there is a division of labour in the hive – the type of bee upon which the mite is riding around the colony on.

For the purpose of this post we’ll consider bees of three ages – newly emerged, nurse bees and foragers. Newly emerged bees (days 1-2 post emergence) clean cells and nurse bees (days 3-11) feed developing larvae. Older bees are involved in wax production (days 12-17) and foraging (>18 days until death).

Logic would dictate that mites would ‘choose’ 2 to associate with bees that bring them into contact with developing larvae of the right age to infest.

Do they?

Hanging around with nurses

Xie et al., (2016)3 assembled artificial colonies containing equal numbers of new bees, nurse bees and foragers, all suitably marked so their age was known. These colonies were provided with a queen, open brood and stores. At the start of the experiment the colonies had 1500 bees and low Varroa levels.

The scientists then introduced 200 ‘phoretic’ mites from another colony 4 and left the colonies for 48 hours. They then age-sorted the bees and harvested all the ‘phoretic’ mites by washing them off in alcohol.

'Phoretic' mites prefer nurse bees

‘Phoretic’ mites prefer nurse bees

On average, ~16% of the nurse bees had ‘phoretic’ mites attached. In contrast, only ~10% of the foragers and ~5% of the new bees had mites. These studies involved seven individual experiments, in two countries and two separate years, using a different source colony for the mites. The statistics are significant.

So mites prefer nurse bees.

Is this ‘simply’ 5 because the nurse bees are more likely to bring the mite into close proximity with a suitably-aged larvae?

Or does associating with, and presumably feeding on, nurse bees have other benefits for the mite?

Mite fecundity and fitness

Fecundity is the reproductive productiveness 6 of an organism.

We’ve obliquely tackled this subject recently. Using an in vitro artificial ‘feed packet’ system, Ramsey and colleagues demonstrated that mites fed on the fat body of bees laid more eggs i.e. they had higher fecundity.

Xie et al., also tested fecundity of ‘phoretic’ mites from newly emerged bees, nurse bees and foragers. They did this by manually harvesting mites after a 3 day ‘phoretic’ phase, adding them to a pre-pupa and then counting the number of progeny female mites 9 days later.

Mite fecundity and fitness

Mite fecundity and fitness

‘Phoretic’ mites from nurse bees exhibited higher fecundity (more female offspring), higher fitness (more mature female offspring) and lower infertility (female mites that did not generate offspring).

So evolution has elegantly resulted in ‘phoretic’ mites associating with the right type of bee to bring them close to developing larvae (upon which they reproduce) and made them better able to reproduce once they get there.

Why do Varroa mites prefer nurse bees?

This is the title of the Xie et al., paper.

Xie et al., sort of answer the question they posed in the title. What they don’t do is explain why ‘phoretic’ mites on nurse bees are more fecund. However, the recent Ramsey paper suggests that this may be because nurse bees have a larger fat body and higher levels of vitellogenin.

If they’re better fed perhaps they produce more viable offspring? 7.

Another known unknown (semiochemicals)

How do mites detect the differences between new, nurse and older bees?

Perhaps they ‘smell’ different?

Mites preferentially infest drone brood because it produces a range of methyl and ethyl esters of straight-chain fatty acids, in particular methyl palmitate.

Similarly, the preference for nurse bees might be explained by their production of another semiochemicals 8. If we could identify this semiochemical it might be possible to create a ‘sponge’ soaked in it that attracted all the mites in the colony.

A bit simplistic, but you get the idea.

In reality, it’s likely that nurse bees are identified by the relative strengths of a range of semiochemicals produced by bees of different ages.

In reality it’s also likely that dropping a ‘sponge’ soaked in eau de nurse bee into the colony could unbalance all sorts of other events in the hive … 🙁

I told you it wasn’t simple.


Rattus norvegicus

Rattus norvegicus

Hanging Around is the fifth track on Rattus norvegicus, the 1977 debut album of one of the finest rock/punk bands of all time, The Stranglers. The album also includes the incomparable Peaches and (Get a) Grip (On yourself), both of which were released as singles.

No more heroes from the same year, but a different album, is also a classic by the same band.

You had to be there … I was 😉

 

 

Pedantically not phoresy

The life cycle of the ectoparasitic mite Varroa destructor essentially consists of two stages. The first is within the capped cell, where reproduction takes place. The second occurs outside the capped cell when the recently-mated female progeny mites matures while riding around the colony attached to a nurse bee.

Almost without exception this second stage is termed the phoretic phase.

It isn’t.

Phoresy

Phoretic is an adjective of the word phoresy. Phoresy is derived from the French phorésie which, in turn, has its etymological origins in the Ancient Greek word φορησις.

And φορησις means being carried.

Which partly explains why the correct definition of the word phoresy is:

An association between two organisms in which one is carried on the body of the other, without being a parasite [OED]

Phoresy has been in use for about a century, with the word phoretic first being recorded in the Annals of the Entomological Society of America (25:79) in 1932:

It is possible, as suggested by Banks (1915), that such young mites are phoretic, being carried about from place to place on the host’s surfaces.

And, no, they weren’t discussing Varroa.

“Without being a parasite”

These are the critical words in the dictionary definition of phoresy which makes the use of the word phoretic incorrect when referring to mites on nurse bees.

Because mites on nurse bees are feeding – or at least a significant proportion 1 of them are.

They are therefore being parasitic and so shouldn’t be described as phoretic.

Om, nom, nom 2

Last week I discussed the recent Samual Ramsey paper presenting studies supporting the feasting of Varroa on the fat body of bees.

In the study they harvested bees from a heavily mite-infested hive and recorded the location on the bee to which the mite was attached.

The majority were attached to the left underside of the abdomen. More specifically, the mite was wedged underneath the third abdominal tergite 3.

What were they doing there? Hiding?

Yes … but let’s have a closer look.

Ramsey and colleagues removed some of the mites and used a scanning electron microscope to examine the attachment point on the bee. Underneath the tergite there is a soft membrane. The imprint of the body of the mite was clearly visible on the membrane.

Varroa feeding location on adult bee

Scanning EM of Varroa feeding location on adult bee

The footpads of the mite were left attached to the membrane (left image, white arrows), straddling an obvious wound where the mouthparts had pierced the membrane (black arrow). Between them, the inverted W shape is presumably the imprint of the lower carapace of the mite.

The close-up image on the right even shows grooves at the wound site consistent with the mouthparts of the mite.

These mites were feeding.

Extraoral digestion

Varroa belongs to the order (a level of classification) Mesostigmata. Most mesostigmatids feed using a process termed extraoral digestion.

Extraoral digestion has also been termed ‘solid-to-liquid’ feeding. It involves the injection of potent hydrolytic enzymes which digest solid tissue, converting it to a semi-solid that can be easily ingested. It can reduce the time needed to feed and it increases the nutrient density of the consumed food.

If Varroa fed on haemolymph it wouldn’t need to use extraoral digestion. Instead it would need all sorts of adaptations to a high volume, low nutrient diet. Varroa doesn’t have these. It has a simple tube-like gut parts of which lack enzymatic activity … implying that digestion occurs elsewhere.

A picture is worth a thousand words

Do the images of feeding mites support the use of extraoral digestion?

EM cross-section of Varroa feeding

EM cross-section of Varroa feeding

The image above 4 shows the cross-section of a Varroa (V), wedged under the tergite (Te), feeding through a hole (arrow in the enlargement on the right) in the membrane (M). The fat body (FB) is immediately underneath the membrane. The scale bar is incorrectly labelled 5.

A close-up of the wound site shows further evidence for extraoral digestion.

Feeding wound at higher magnification

Feeding wound at higher magnification

Beneath the wound site (C, arrow) are remnants of fat body cells (white arrow) and bacteria (black arrow; of two types, shown in D). A closer look still at the remnants of the fat body (E and F) shows cell nuclear debris (blue arrows) and lipid droplets (red arrows).

These images are entirely consistent with extraoral digestion of fat body tissue by feeding Varroa. The presence of bacteria near the wound suggests that bacterial infection may result from Varroa feeding, possibly further contributing to disease in bees.

So, pedantically it’s not phoresy

So-called phoretic mites, unless they’re on the thorax or head of the bee, are not really phoretic. They are being carried about, but they are also likely feeding. By definition that excludes them from being phoretic.

Instead they are ectoparasites of adult bees.

What are the chances that beekeepers will stop using the term phoretic?

Slim to none I’d predict 6.

And, of course, it doesn’t really matter what the correct term for them is.

What’s more important is that beekeepers remember that it’s at this stage that mites are susceptible to all miticides.

The June gap

But it’s also worth thinking about the potential impact of brood breaks.

During brood breaks all the mites in the colony must be ‘phoretic’.

Generally, the majority of the mites in a hive are in capped cells. Depending upon the stage of the season, the egg-laying rate of the queen and other factors, up to 90% of the mites are associated with developing pupae.

But as the laying rate dwindles more and more mites are released from cells and become ‘phoretic’, unable to find a suitable late-stage larva to infest.

And which bees do the mites associate with?

Nurse bees primarily, for reasons I’ll discuss in the future. But – spoiler alert – one of the reasons is likely to be that they have a larger fat body.

So, a mid-season brood break (e.g. the ‘June gap’) is likely to result in lots more nurse bees becoming both the carriers and the dinner of the mite population.

Some or many of the nurse bee cohort may perish, perhaps from damage to the fat body or from the viruses acquired from the mite. However, bees exhibit phenotypic plasticity, meaning that older bees can revert to being nurse bees when the queen starts laying again.

Late season brood breaks

In late summer mite levels are usually at their highest in the hive. A brood break occurring now will release a very large number of mites to parasitise the adult bee population.

Presumably these mites select the bees best able to support them 7.

And which bees are these? The nurse bees of course. But it’s also worth remembering that there are key physiological similarities between nurse bees and winter bees. Both have low levels of juvenile hormone and high levels of vitellogenin (stored in the fat body).

So I’d bet that the ‘phoretic’ mites during a late season brood break would also preferentially associate with any early-produced winter bees.

Furthermore, once the queen starts laying again – perhaps in early/mid-autumn – the winter bees being produced would be subjected to the double-whammy of high levels of mite infestation and potential damage from ‘phoretic’ mites.

Practical considerations

More work is required to model or actually measure the impact of late season brood breaks, high levels of ‘phoretic’ mites, nurse bee numbers and winter bee development.

Compare two colonies of a similar size with a similar mite load, treated at the same time in early autumn with an appropriate miticide. If one of them experienced a late summer brood break (pre-treatment) and consequent high levels of ‘phoretic’ mites, does this reduce the chances of the colony surviving overwinter?

Who knows? Lots and lots of variables …

Fundamentally, it remains important to treat colonies early enough to protect the winter bee population. You’ve heard this from me before and you’ll hear it again.

However, it’s something to think about and I can see ways in which it might influence the strategy and timing of mite control used. I’ll return to this sometime in the future.


 

Chewin’ the fat

A little over a year ago reports started to circulate of a study showing that Varroa feed on the fat body of bees rather than on haemolymph.

Having worked in Glasgow through the early noughties the title of this post was a no-brainer and an outline draft was written in December 2017. However, the peer-reviewed paper wasn’t published until last month, so it’s only now we’ve got the chance to judge the study and consider its implications.

Varroa feed on hameolymph, right?

Historically this was the accepted dogma. However, the experimental data supporting this conclusion – based upon labelling bees with radioactive isotopes and seeing what the mites acquired after feeding – was really not definitive. The experiments had been done in the 1970’s and the specificity of the labelling was a bit dubious. In addition, during the intervening period scientists had determined that, unlike vertebrate blood which is rich in cells and nutrients 1, haemolymph has little of either and is actually a pretty lousy food source.

In addition, and somewhat more circumstantially, Varroa control using chemotherapeutics fed to bees (and subsequently taken up by the mite during feeding) had been relatively disappointing.

Perhaps these chemicals weren’t getting to the right tissues of the bee?

Perhaps Varroa don’t feed on haemolymph after all?

The Ramsey study

This new study reports three independent experiments that, together, indicate that Varroa actually feed on the fat body of bees, rather than on haemolymph. The paper is so-called ‘open access’, so anyone can access it and therefore I’ll just provide a synopsis of the important bits.

The questions Samual Ramsey and colleagues attempted to answer were:

  1. Where on the bee do mites feed? Is it primarily or exclusively near the fat body?
  2. When Varroa feeds, what host tissues are ingested?
  3. What sort of diet is required to maintain Varroa and allow their reproduction in vitro2.

Location, location, location

The authors counted phoretic mites on 104 bees. Over 95% of them were located on the underside of the body, predominantly on the left side of the bee, under the tergite or sternite3 on the third metasomal segment (i.e. the second visible segment of the abdomen).

Mite location on nurse bees

Mite location on nurse bees

This position is consistent with feeding on the fat body tissues which are most abundant under the inner ventral surface of the metasoma.

Seeing red

Bees were fed with Nile red, a lipophilic fluorescent stain that preferentially accumulates in the fat body. They co-fed bees with uranine, a differently coloured fluorophore that accumulates in the haemolymph. They then allowed mites to feed on the fluorescently labelled bees and subsequently photographed the mites under fluorescent light.

The rationale here was straightforward. If the mites fed on the fat body they would stain red due to taking up the Nile red stain.

Mites visualised after feeding on fluorescently labelled bees

Mites visualised after feeding on fluorescently labelled bees

Which they did.

It was notable that the red stain predominantly accumulated in the rectum and gut of the mite (image O above). The authors conducted all sorts of controls to confirm that the stains actually stained what they were supposed to – you can view these in the paper.

Babies!

In the final part of the study the authors maintained mites in vitro (in an incubator), feeding them on a diet containing increasing amounts of fat body or haemolymph. These are tricky experiments and in some way the least satisfactory part of the study.

Two results suggest that fat body was beneficial or essential to the mites. Firstly, only mites that had 50% or more fat body in the diet survived for 7 days. Secondly, there was a dose response to the amount of fat body in the diet and fecundity. Mites on a 100% fat body diet exhibited 40% fecundity, the highest level observed in the study.

What can we conclude from the Ramsey study

Of the three experiments presented, the Nile red fat body stain uptake by mites is reasonably compelling.

The feeding position study is essentially correlative, but there could be other interpretations of the data. For example, that location on the bee might be the least accessible to a ‘grooming’ bee. Perhaps it’s a survival mechanism?

Survival and fecundity in in vitro studies wasn’t great. However, in defence of the authors, fecundity of mites under natural conditions can be as low as 40% and is not higher than 80%. Not all mites have baby mites. Thankfully.

Only 20% of the mites survived one week under in vitro conditions, even on a 100% fat body diet. In contrast, mites fed haemolymph alone died within 48 hours. This poor level of survival was surprising and suggests other essential components of the diet were probably missing.

Other published studies have shown reasonable survival of Varroa for at least 3 days, with at least one report of mites surviving on flowers for up to 7 days. I’m also aware that other laboratories can maintain mites in vitro for longer than 7 days without using any honey bee-derived components in the diet.

Hang on … what is the fat body anyway?

The fat body is multi-functional. It has been compared to the vertebrate liver and adipose tissue. It acts as a major organ for nutrient storage, energy metabolism and detoxification of things like pesticides.

Vitellogenin made by and stored in the fat body reduces oxidative stress and is associated with extending the longevity of overwintering bees. The fat body also has critical roles in metamorphosis.

So, not only multi-functional, but also very important.

Significance of the results … is this a game changer?

This paper has been discussed online as a ‘game changer’. That’s probably a bit strong. Whilst the fluorescent stain uptake study is reasonably convincing it must be remembered that it was conducted on adult bees.

Do mites on pupae also feast on the fat body?

This will have to be determined in the future. It’s a more difficult experiment of course.

The other two studies, and a number of additional small observations I’ve not discussed here, are certainly supportive, but not alone hugely convincing. The in vitro study in particular will be interesting to compare with (currently unpublished) studies from other laboratories that do not use honey bee fat bodies in their mite feeding and maintenance diet.

Practical matters

Does it matter what part of the bee the mite feeds on?

Clearly it does for the mite, but what about the beekeeper?

I think this study is significant for the beekeeper for two reasons – the first will only be relevant if and when lipophilic miticides are developed, the second matters right now.

  1. Strategies are being developed to add highly specific miticides to the diet of bees which are then delivered to Varroa when the mite feeds. To date, these have been rather underwhelming in their performance. If Ramsey is right, modification of these miticides to make them lipophilic (like the Nile red fluorphore) will concentrate them in precisely the right place to ensure the mites get a lethal dose.
  2. A key product of the fat body is vitellogenin. The long-lived overwintering bees have high levels of vitellogenin. Mites feeding on, and depleting, the fat body would be expected to result in reduced vitellogenin levels in the bee 4. This would explain why high Varroa levels are associated with reduced longevity of winter bees and consequently increased overwintering colony losses.

The most important take home message

To prevent mites that feed on fat bodies from damaging vitellogenin production miticides have to be used early enough to protect the winter bees.

In the paper Ramsey makes the statement:

Simple reduction of mite loads late in the season to decrease the overwinter parasite load may not be enough, as it still allows for the mites to damage tissue critical to the process of overwintering …

Instead …

A treatment schedule that includes treatment in late summer or early fall before mites can significantly damage fat body in developing winter bees would likely be more effective.

Which is precisely the point I’ve made previously about treating early enough to protect winter bees.

What the Ramsey paper adds is the piece of the jigsaw possibly explaining why late summer treatment is so important.


Colophon

Chewin’ the Fat was a four-series Scottish comedy sketch show. It was broadcast from 1999 to 2002, with further Hogmanay specials until 2005. The show had a recurring cast of characters and sketches including The Big ManThe Banter BoysThe Lighthouse KeepersBallistic Bob and Taysiders in Space.

Gonna no' dae that

Gonna no’ dae that – The Lighthouse Keepers

Chewin’ the Fat was filmed in and around Glasgow (where I worked at the time) and the characters parodied a range of local ‘types’ … pretentious Kelvinsiders, Glaswegian gangsters, narcissistic golfers, The man from Kilmacolm, and shellsuit-wearing, chain-smoking, hard-drinking Glaswegian neds.

It was a bit rude and definitely an acquired taste. Without subtitles, some of the scenes would probably have been unintelligible south of the border.

Mites equal viruses

Healthy bees are happy bees 🙂

Sounds good doesn’t it?

Actually, there’s no evidence that bees display or perceive most of the emotions often attributed to them 1.

Happy? Who knows? But certainly not healthy ...

Happy? Who knows? But certainly not healthy …

A more accurate statement might be “Healthy bees are more productive, they are less likely to die overwinter, less likely to be robbed out by wasps or neighbouring strong colonies and their parasites and pathogens cannot threaten the health of other honey bee colonies or, through so-called-pathogen overspill, the health of other pollinators.”

More accurate?

Yes … but it doesn’t exactly trip off the tongue 😉

Whether it makes the bees happy or not, beekeepers have a responsibility to look after the health of their livestock. This includes controlling Varroa numbers to reduce the levels of pathogenic viruses in the hive.

How well are virus levels controlled if mite levels are reduced?

I’ll get to that in due course …

Midwinter mite massacre

The 2018 autumn was relatively mild through until mid/late November. In the absence of very early frosts colonies continued rearing brood.

We opened colonies in mid-November (for work) and found sealed brood, though it was clear that the laying rate of the queen was much-reduced.

These are ideal conditions for residual mite replication. Any mites that escaped the late summer/early autumn treatment (the ideal time to treat to protect the overwintering bees) continue to replicate, resulting in the colony starting the following season with a disappointingly high level of mites.

I’ve noted before that midwinter mite levels are paradoxically higher if you treat early enough in the autumn to protect the all-important winter bees.

Consequently, to start the year with minimal mite levels, I treat in midwinter with a trickled or vaporised oxalic acid-containing (OA) treatment.

A combination of colder weather (hard frosts in late November) and brood temperature measurements 2 indicated mid-December was a good time to treat.

Midwinter mite massacre

Midwinter mite massacre

18th December

In one of my apiaries ten colonies were treated. Some were definitely broodless (based upon Arnia hive monitoring). Others may have had brood, but colonies were not routinely checked.

Over the four day period after vaporising these ten colonies dropped a total of 92 mites. More than 50% of these were from just one double-brooded colony. Overwintering nucs 3 dropped no mites at all in the 12 days following treatment.

This was very encouraging. These are lower midwinter mite levels than I’ve seen since returning to Scotland in 2015.

The one colony with ‘high’ mite levels received two further treatments (on the 22nd and 27th) in an attempt to minimise the mite levels for the start of the season. Going by the strength of the colony and the debris on the Varroa tray it was presumed that this colony was still rearing brood.

Mite drop following the third treatment was negligible 4.

Why are mite levels so low?

I think it’s a combination of:

  • Luck
  • Use of natural, organic, bee-centric and biodynamic beekeeping methods
  • Varroa-resistant bees
  • Very tight control of mite numbers in the 2017/18 season, primarily by correctly timing the winter and the late-season autumn treatments. This is simply good colony management. Anyone can achieve this.
  • A brood break midseason and/or a broodless period when splitting colonies (both give opportunities for more phoretic mites to be lost through grooming). Undoubtedly beneficial but season-dependent. I’ll be discussing ways to exploit these events in posts next year.
  • A low density of beekeepers in Fife, so relatively little drifting or robbing of poorly managed colonies from neighbouring apiaries. Geography-dependent. Much easier in Fife than Warwickshire … and easier still in Lochaber.

And what do less mites mean?

Varroa is a threat to bee health because it transmits pathogenic viruses when feeding on developing pupae.

The most important of these viruses is deformed wing virus (DWV).

Generally, the higher the level of infestation with mites, the higher the viral load 5. This has been repeatedly demonstrated by studies from researchers working in the UK, Europe and the USA.

It is well-established that colonies with high viral loads have an increased chance of dying overwinter, due to the decreased longevity of bees infected with high levels of virus.

DWV symptoms

DWV symptoms

In our work apiaries we regularly measure DWV levels. For routine screening our limit of detection is around 1,000 viruses per bee.

We don’t actually count the viruses. They’re too small to see without an electron microscope 6.

Instead, we quantify the amount of the virus genetic material present 7, compare it to a set of standards and express it as ‘genome equivalents (GE)’.

Many of the bees tested this year contained ~103 (i.e. 1000) GE, which is extremely low. Bees from Varroa-free regions (e.g. Colonsay) carry similar levels of DWV.

Most of our colonies were at or close to this level of virus much of the 2018 season. This is 100-1,000 times lower than we often see even in apparently perfectly healthy colonies in other years or other apiaries.

For comparison, using the same assay we usually detect about 1010 (ten billion) DWV GE per bee in symptomatic adult bees from heavily mite-infested colonies.

So, less mites means less viruses which means healthier bees 🙂

And they might even be happier bees 😉

And your point is?

It’s worth remembering that the purpose of treating a colony with miticides is to reduce the transmission of viruses between bees. This transmission results in the amplification of DWV. This is why the timing of treatments is so important.

Yes, it’s always good to slaughter a few (or a few thousand 🙂 ) mites. However, far better massacre them when you need to protect particular populations of bees.

This includes the overwintering bees, raised in September, that get the colony through to the Spring.

Remember also that it ‘takes bees to make bees’ i.e. the rearing of new brood requires bees. Therefore strong colony build-up in Spring requires healthy workers rearing healthy brood.

This is why it’s important to minimise mite levels in midwinter when colonies are broodless.

What do most beekeepers do?

Fifteen months ago I published a post on the preparation of oxalic acid solutions for trickling colonies in midwinter.

Whatever the vapoholics on the online forums claim, trickling remains the easiest, quickest and least expensive way to treat colonies in midwinter 8.

The best time to treat in the winter is when the colony is broodless. Here in Fife, and often elsewhere, I believe that this usually occurs earlier in the winter than many beekeepers treat (if it happens at all … or if they treat at all).

I usually treat between the end of the third week in November and mid-December, at the end of the first extended cold period.

Oxalic acid preparation recipe page views

Oxalic acid preparation recipe page views

Looking at the page views for these oxalic acid recipes it looks as though many beekeepers treat after Christmas 9 … which may be suboptimal if colonies had a broodless period and now started rearing brood again.

Mine have.

This winter has been quite mild (at least at the time of writing) so there may yet be opportunities to treat really effectively during a broodless period.

Or the chance may have gone …


 

Know your enemy

What less appropriate time is there, as we enter the festive season of goodwill, to provide a brief account of the incestuous and disease-riddled life cycle of the Varroa mite?

Happy Christmas 🙂

Scanning electron micrograph of Varroa destructor

Scanning electron micrograph of Varroa destructor

Varroa is the biggest enemy of bees, beekeepers and beekeeping. During the replication cycle the mite transfers a smorgasbord of viruses to developing pupae. One of these viruses, deformed wing virus (DWV), although well-tolerated in the absence of Varroa 1replicates to devastatingly high levels and is pathogenic when transferred by the mite.

Without colony management methods to control Varroa, mite and virus replication will eventually kill the colony.

I’ve written extensively on ways to control Varroa. Most of these have focused on early autumn and midwinter treatment regimes. However, next season I’m hoping to discuss some alternative strategies and will need to reference aspects of the life cycle of Varroa … hence this post.

What is Varroa?

Varroa destructor is a distant relative of spiders, both being members of the class Arachnida … the joint-legged invertebrates (arthropods). It was originally (and remains) an external parasite (ectoparasite) of Apis cerana (the Eastern honey bee) and – following cross-species transfer a century or so ago – Apis mellifera, ‘our’ Western honey bee.

Apis cerana, having co-evolved with Varroa, has a number of strategies to minimise the detrimental consequences of being parasitised by the mite.

Apis mellifera doesn’t. Simple as that 2.

One hundred years is the blink of an eye in evolutionary terms and, whilst there are bees that have partial solutions – largely behavioural (small colonies and very swarmy) – they’re probably unable to collect meaningful amounts of honey 3.

Varroa-resistant honey bees will probably evolve (as much as anything is predictable in evolution) but not in my time as a beekeeper … or possibly not until Voyager 2 leaves the Oort Cloud 4.

And there’s no guarantee they’ll be any use whatsoever for beekeeping …

The replication cycle of Varroa

Varroa has no free-living stage during the life-cycle. The adult mated female mite exhibits two distinct phases during the life-cycle. It has a phoretic phase on adult bees and a reproductive phase within sealed (‘capped’) worker and drone brood cells. Male mites only ever exist within sealed brood cells.

I’m going to discuss phoretic mites in a separate post. I’ll concentrate here on the replication cycle.

The mated female mite enters a cell 15-50 hours before brood capping. Drone brood is chosen preferentially (at ~10-fold greater rates than worker brood) and entered earlier. Depending upon the time of the season and the levels of mites and brood, up to 70-90% of mites in the colony occupy capped cells.

The first egg is laid ~70 hours after cell capping. This egg is unfertilized and develops into a haploid male mite. Subsequent eggs are fertilised, diploid, and so develop into female mites. These are laid at ~30 hour intervals.

The replication cycle of Varroa

The replication cycle of Varroa

Worker and drone brood take different times to develop. Therefore a typical reproductive cycle involves five eggs being laid in worker brood and six in drone brood. Not all of these eggs mature, their development being curtailed by the bee emerging as an adult.

There are all sorts of developmental stages involved in getting from an egg to a mature unfertilised mite, but these are not important in terms of the overall outcome. Mite-geeks love this sort of detail 5, but we need to cut to the chase …

Keeping it in the family

The foundress ‘mother’ mite and her progeny all share a single feeding hole through the cuticle of the developing pupa.

What a lovely scene of family ‘togetherness’. 

Male and female mites take 6.6 and 5.8 days respectively to develop to sexual maturity. Therefore the male mite reaches sexual maturity before the first of his sisters.

He then lurks around the attractive-sounding “faecal accumulation site” and mates with each of the (sister) females in turn.

What a little charmer 😉

Male mites are short lived and the eclosion of the adult worker or drone curtails further mating activity, releasing the foundress mite and the mated mature daughters 6.

Reproductive rate (mites per cell)

The three day difference in the duration of worker and drone development means that more mites are produced from drone cells than worker cells. Depending on conditions the reproductive rate is 1.3 – 1.45 in worker brood and 2.2 – 2.6 in drone brood.

Remember that the foundress is also released from the cell. She can go on to initiate one or two further reproductive cycles (or up to 7 in vitro). Consequently, the average yield of mature, mated female mites from worker and drone cells is a fraction over 2 and 3 respectively.

Before entering a fresh cell containing a late stage (5th instar) larva the newly-mated mites need to mature. They do this during the phoretic phase which lasts 5-11 days. Therefore the full replication cycle of the mite probably takes a minimum of about 17 days.

Exponential growth

Two to three mites per infested cell doesn’t sound very much. However, under ideal conditions this leads to exponential growth of the mite population in the colony. Assuming 10 reproductive cycles in 6 months, a single mite would generate a population of >1,000 in worker brood and >59,000 in drone brood 7.

Fortunately (for our bees, not for the mites), ideal conditions don’t actually occur in reality.

Lots of things contribute to the reduction in reproductive potential. For example, only 60% of male mites achieve sexual maturity due to developmental mortality, drone brood is only available at certain times in the season, brood breaks interrupt the availability of any suitable brood and grooming helps rid adult bees of phoretic mites.

Out, damn'd mite ...

Out, damn’d mite …

However, these reductions aren’t enough. Without proper management mite levels still reach dangerously high levels, threatening the long-term viability of the colony.

In the next few months I will discuss some additional opportunities for reducing the mite population.

In the meantime, as we reach the winter solstice, colonies in temperate regions may well be broodless and – as emphasised last week – this is an ideal time to apply a midwinter oxalic acid-containing treatment. This will effectively reduce mite levels for the start of the coming season.

Happy Christmas … unless you’re a mite 😉


Colophon

Today is the winter solstice in the Northern hemisphere. This is actually the precise time when the Earth’s Northern pole has its maximum tilt away from the Sun. However, the term is usually used for the day with the shortest period of daylight and the longest period of night. In Fife, sunrise is at 08.44 and sunset at 15.37, meaning the day length is 6 hours and 53 minutes long.

With increasing day length queens will start laying again … but there’s a long way to go until winter is over.

 

The eyes have it

We’re entering the not beekeeping end-of-season phase of the beekeeping year. There’s been a marked reduction in visitor numbers to ‘The Apiarist’ over the last few weeks and – with the weather gradually deteriorating1 – the ‘shack nasties‘ are starting to develop. The online forums (fora?) are filled with increasingly bad-tempered arguments discussions and it might be too soon to be thinking about 2019 (it isn’t).

Wasted words

So, rather than write a series of erudite, well-argued, coherent, logical and persuasive posts about evolution of Varroa resistance in Apis mellifera, or rational mid-season mite-management strategies, or the 75:25 rule for queen and stock-improvement, or an exhaustive review of Swienty vs. Abelo poly Nationals …

Some chance !2

… I’m instead going to spend the next few weeks on a variety of odds and ends. Some interesting and amusing science, some ‘teasers’ on grow-your-own-denim-knit-your-own-yoghurt beekeeping, an introduction to why people keep bees and why they shouldn’t and a long and apologetic explanation of where the site disappeared to when I tried to move it to another server.

Actually, of these only the first exists (see below). The last I hope not to use, though I will be switching servers to accommodate changes in security and to speed things up and to introduce site-wide intrusive advertising and a subscription model to fund my seemingly-unstoppable purchasing of essential beekeeping stuff from Brian at Thorne’s of Newburgh.

Oops.

The eyes have it

The eyesight of bees is remarkable.

Actually, eyesight alone is not enough. It’s the combination of eyesight with the neuronal processing of the received images that’s truly remarkable.

Remember that the brain of a bee is about 1mm3 and contains about one million neurones 3. With this brain the bee is able to undertake a series of complex mental tasks involving learning and memory, image processing and visual generalisation.

Bees soon learn that particular flowers yield lots of pollen or nectar. They can return to them time and again, recognising them at relatively short distances from their appearance. How do they determine that other flowers – of different shapes, sizes or colours – might also have valuable pollen or nectar? What about tree flowers that have a different appearance again?

It turns out that bees are generalists, at least where flowers are concerned. They recognise things that are flower-like. They have evolved to associate reward (pollen, nectar) with things that have the appearance of flowers.

More general generalists?

Do they only have the ability to identify flower-like ‘things’. Are bees generalists when just identifying flowers and flower-like things, albeit of different colours, sizes and shapes? Is there some sort of hardwiring in the brain of the bee that has evolved this exquisite combination of flower-recognising sensitivity and flexibility?

Alternatively, perhaps bees have a more adaptable image processing capability? For example, we know through simple experiments that bees can rapidly learn to associate very unflower-like shapes with a syrup ‘reward’.

You can train bees to repeatedly return to a distinctively coloured/shaped item with a syrup reward. Over short distances you can move the item and the bees return to the new location, with the final approach being guided by vision, pattern recognition and associated image processing.

The item doesn’t need to look much like a flower.

They can also identify ‘diamond-shaped things’ (again, for example, other shapes are available at a bee lab near you) of a different colour, or even no colour, to those they’ve been trained on.

Shape recognition by bees

Shape recognition by bees …

This suggests that – at least for simple ‘not-flower’ shapes like these – a degree of generalisation is still possible.

But bees operate in a very busy and variable environment filled with shapes and colours that form wildly variable complex images. Generalisation might well be a problem with all of this variation and complexity.

Facial recognition

Bees are good at discriminating between images of simple shapes. How good are they are at recognising the sorts of complex shapes and images that are found in the environment?

What about something that our bees see every week?

Something they might associate with disruption and/or reward?

Like your face …  😀

It turns out that bees can distinguish between faces. Very well. When trained to a syrup reward on one face (top left in image below), they can distinguish it from a different face (second row) about 80% of the time (graph).

Facial recognition by bees ...

Facial recognition by bees …

There’s a distinct possibility your bees recognise you. An interesting twist on the comment many non-beekeepers make about whether we can identify ‘our’ bees.

However, bees trained to recognise a face the ‘right way up’ failed to identify the face if it was inverted (column v above).

Don’t do your hive inspections standing on your head.

Complex image generalisation

Bees can certainly discriminate well between complex images like faces. Are they also able to generalise when it comes to complex image analysis?

For example, could you train bees to associate reward with a range of female faces? Then challenge them with discriminating between a pair of new (never seen before) male and female faces? Would they pick the female face significantly more than 50% of the time?

That’s a pretty tough test. Without the labels how well do you cope with this training set?

Faces

Faces

OK, if that’s too hard, how about an analysis of image generalisation based upon the style of the image?

Humans are pretty good at this sort of thing. We can easily discriminate between the Impressionist painters (e.g. Degas, Monet, Manet, Renoir) and those in the early 20th Century Cubist art movement (e.g. Picasso, Metzinger, Braque, Gleizes, Léger).

For example, is the Picasso (below) Cubist or Impressionist?

We can tell … can the bees?

Monet or Picasso?

Wu and colleagues4 recently attempted to answer this question.

They took pairs of paintings matched for luminance, colour and spatial frequency information, one by Monet (Impressionist) and one by Picasso (Cubist). Bees were trained to associate either the Monet or the Picasso with a syrup reward.

When subsequently tested, bees were able to easily identify the painting they had been trained on from one by the other artist. After 30 training blocks bees made the correct choice about 75% of the time … approximately the same accuracy with which they identify faces (above).

This is not fundamentally a different experiment to the face recognition study as both involved the discrimination between just one complex image and another.

Monets or Picassos?

Using five different pairs of luminance, colour and spatial frequency-matched paintings from Monet and Picasso – with five days of training – they demonstrated that bees could simultaneously discriminate between them up to 75% of the time.

The more training the bees received, the better they were at picking the correct painting each time.

Impressionist or Cubist?

Having trained the bees on multiple Picasso or Monet paintings they then challenged them with new (to the bees … you’ll appreciate that these artists no longer produce new work 😉 ) paintings by the same artists.

Could bees that were able to discriminate between The Cliff at Étretat after the Storm (Monet) from Le Rêve (Picasso) and between Water Lilies and the Japanese bridge (Monet) from Girl before a Mirror (Picasso) and three other pairs correctly select a previously unseen Picasso or Monet?

In all honesty, not very well 🙁

The statistics are poor. For one of the novel pairs tested it appeared as though the bees could discriminate as well as they could one of the training pairs. However, the most positive statement that could be made by the authors was “Notably, for both groups the percentage of correct choices for novel pairs was above chance (i.e., above 50 %) in six out of the eight tests, indicating that a weak generalization may have occurred”.

Underwhelming … in this sort of science the stats wins every time, and this type of statement isn’t very compelling.

Finally, the scientists repeated the entire training regime with greyscale versions of the same training pairs of images. With this training, generalization to the novel pairs was quite a bit better with “only marginal or no significant difference between training pairs and most novel pairs”.

Better, but still not really statistically compelling. However, don’t underestimate the complexity of the task. The results showed that insects with a sesame-seed-sized brain could often discriminate between previously unseen Cubist or Impressionist paintings after a few days training on only 5 pairs of paintings of the same style.

That’s remarkable.

Bird brains

Pigeons live in the same visually complex environment as bees. They have to undertake similar visually demanding tasks during foraging. They can discriminate between Monets and Picassos. They can correctly (and statistically convincingly) determine whether a new Monet or Picasso is more Impressionist-like or Cubist-like.

In addition, when challenged with other paintings of similar styles by different artists (e.g. a Degas or a Braque), pigeons can again generalise in their selection of Cubist or Impressionist 5.

However, to achieve this remarkable visual feat, pigeons need to be trained to hundreds of exemplar paintings over many, many weeks.

Could bees do as well if trained for the same period?

We don’t know.

And we’re unlikely to find out as the lifespan of a worker bee is probably too short 🙁


Colophon

This post was written as the political fallout of the draft Brexit deal was occupying 110% of the news. By the time it appears online it’s not clear the UK will have a Prime Minister or even a functioning Government.

Be that as it may, there will be a Parliamentary vote on it.

Historically, there is a division of the assembly into those that support the motion (the ‘ayes’ i.e. ‘yes’) and those that do not (the ‘noes’). Once the vote is taken – typically by members of parliament traipsing into the appropriate division lobby – the Speaker counts the votes and announces The Ayes have it … assuming the motion was supported.

Considering the timing, a pun on The Ayes have it seemed appropriate.

Resistance is not futile

Apivar ...

Apivar …

Amitraz-containing miticides are sold in the UK as Apivar and Apitraz.

Until recently they were only available with a veterinary prescription. I expect – though I have not yet seen data to support this – that their usage in the UK will increase now they are off-prescription. These miticides are now widely available and so there is greater opportunity to use – and misuse – them.

If you’re using Apivar 1 for the first time this year you will soon have to remove the strips from the hive.

That’s assuming you started treating early enough to protect the all-important winter bees from Varroa and its deadly viral payload.

This post is a reminder to remove the strips at the right time. The alternative – leaving them in place until the first Spring inspections – risks help the development of resistance to amitraz, so further reducing our opportunity to control mites effectively.

Leave and let die

Without careful integrated pest management (IPM) 2 Varroa levels build up in the hive. Varroa transmits viruses – most important of which is deformed wing virus (DWV) – to developing pupae. High levels of DWV either kills the pupa or results in emergence with or without significant developmental defects. Even those bees that are apparently normally developed have a reduced lifespan 3.

Winter bees with a reduced lifespan prevent the colony from surviving through the winter until the queen starts laying again. I’ve also proposed recently that high levels of DWV, and the resulting increased rate of winter bee die-offs, probably accounts for some cases of isolation starvation.

So … intervention is needed to reduce mite levels, protect your bees and save your colonies.

Follow the instructions!

Apivar is one solution to reduce mite levels. It is an easy-to-apply chemical treatment that is very effective in reducing the Varroa load by ~95%. For a National hive it is applied as two polymer strips, each containing 500mg of slow-release Amitraz. Strips are hung between brood frames for 6-10 weeks and – for maximum efficacy – should be scratched with a hive tool and repositioned half way through the treatment period.

Amitraz

Amitraz …

Unlike some other miticides (e.g. Apiguard and MAQS) there are no temperature restrictions for Apivar usage. The colony does not need to be broodless (a requirement for trickled oxalic acid-based treatments) as the treatment period covers multiple brood cycles.

Other than not using it with supers present the only contraindication for Apivar is to not use it if Amitraz-resistant mites are present.

How does resistance develop?

When discussing parasites and pathogens, resistance 4 is a consequence of two things:

  1. A selective pressure that kills the pathogen
  2. A population which exhibits genetic diversity

The selective pressure could be anything … heat for example, antibiotics prescribed by your GP, an antiviral against HIV or – of relevance here – Apivar against Varroa.

Killing – at the population level – is not absolute. Some individuals within the population survive longer than others. They could be exposed to a slightly lower dose, or be located in a protected niche for example. However, treat for long enough and the majority will be killed.

But there’s more …

Pathogen populations are not genetically invariant. Actually, many are quite diverse and have replication cycles that – deliberately 5 – generate diversity.

Therefore some pathogens are genetically slightly less resistant and some are genetically slightly more resistant to a selective pressure. We can ignore the former as they’ll rapidly be killed off … but we must be concerned about the more resistant ones.

Keep taking the pills

All of this is a ‘numbers game’, better represented with graphs and equations. However, the take-home message is simple … to effectively control a pathogen you need to treat for long enough and with a high enough dose to kill the vast majority of the population.

That’s why you’re encouraged to “complete the course” of antibiotics … or to remove the Apivar strips after 10 weeks and not leave them in over the winter.

Because both courses of action result in selection of more resistant pathogens.

If you stop taking antibiotics too soon, you won’t have treated for long enough and with a high enough dose. You end up selecting for the more genetically resistant pathogens.

Similarly, if you leave Apivar strips in overwinter you’ll be “treating” the remaining mites 6 with a lower dose of the miticide, which is an ideal situation to favour the growth of the slightly more genetically resistant mites.

How does Amitraz resistance develop?

Resistance to Amitraz in Varroa is well documented. It’s been described in a number of countries including the USA and Europe, Mexico and Argentina 7. Generally resistance is defined in terms of a reduced level of mite killing, or – in laboratory experiments – an increased dose required to kill a certain proportion of mites.

However, I’m unaware of any studies defining the genetic basis of Amitraz resistance in Varroa.

But Amitraz is a widely-used acaricide 8 and the genetic basis of resistance in cattle ticks is well understood. In these, ticks resistant to Amitraz carry a mutation in the RMβAOR gene 9.

What 10 is the RMβAOR gene?

I’m glad you asked 😉

This gene encodes the β-adrenergic octopamine receptor protein and readers with good memories will recall that this is one of the targets that Amitraz binds to and inactivates 11.

If the protein carries a mutation the Amitraz cannot bind to it and so the mite – or more correctly the tick as it’s yet to be formally demonstrated in mites – is therefore resistant.

(Bad) practical beekeeping

What does all this mean in terms of practical beekeeping?

It means use the correct number of Apivar strips for the colony and leave them in for the right length of time.

Do not …

  • Use one strip on a full colony mid-season to ‘knock back the mites a bit’ 
  • Re-use the strips in another colony (yes really!)
  • Use improperly stored strips (or out of date strips) in which the effective Amitraz dose is reduced

I’ve heard examples of these types of misuse this season. All increase the chance of selecting for Amitraz-resitant mites.

And (the real reason for posting this at this time of year) …

  • Do not leave the strips you added in late summer in the colony throughout the winter

Removing the strips takes seconds. Prize off the crownboard, grab the tab projecting above the top bars, gently withdraw the strip and close the hive up again.

Finally, because of the incestuous lifestyle 12 of Varroa the genetic diversity (and therefore potential presence of more resistant mites) in the population is likely to be increased by the high mite levels that prevail late in the season.

All the more reason to use the effective treatments we currently have in a way that helps ensure they remain effective.


Colophon

Resistance is futile

Resistance is futile

Resistance is futile is the title of a 2018 album by the Welsh rock band the Manic Street Preachers.

More specifically, in the context of this post, it was the phrase routinely used by the Borg – the alien cyborgs sharing a collective mind – in the Star Trek franchise. Borgs rarely speak, but when they do they usually include this phrase. For example “We are the Borg. Lower your shields and surrender your ships. We will add your biological and technological distinctiveness to our own. Your culture will adapt to service us. Resistance is futile.” The warning about resistance being futile was usually accompanied by the threat that the target would be assimilated”.

I’d started writing this post using the title ‘Resistance is futile’ but realised late on that – as far as Varroa are concerned – resistance is anything but futile 13.

Resistance – to miticides – gives Varroa a reason to live. Literally.

Let’s not help them 🙂

Ouch, that hurt

If you keep bees you’ll inevitably get stung.

Not necessarily often and not necessarily badly, but getting stung goes with the territory.

You’ll probably get stung more often if your bees are stroppy, or if you are clumsy. But even if you’re careful and the bees are calm there’s always the chance of being stung.

I moved a very feisty colony late one evening last week 1. The hive was sealed, moved and re-located to an out apiary. Knowing they were, er, rather temperamental I let them settle for 15 minutes, then gently lifted the entrance block.

Out they boiled … as I beat a very hasty retreat 🙁

I thought I’d got away with it, but driving home 20 minutes later I was stung on the ankle by a stowaway in my boot.

Ouch! That hurt.

I’ve only been stung a few times all season. Most didn’t hurt much at the time and were forgotten within minutes. That sting on the ankle hurt like hell and was sore for a further 48 hours.

Why does it hurt when you’re stung? Furthermore, assuming stings are inevitable, which parts of the body hurt more when stung … and so deserve additional protection?

Why do bee stings hurt?

The honey bee sting is a hollow barbed tube used to deliver the venom. About 50% of bee venom by weight is the small protein mellitin.

It’s fair to say that mellitin is small but potent. It’s only 26 amino acids 2 long and forms a tetramer in aqueous solution. The ‘noughts and crosses’ shape it adopts hides the hydrophobic parts of the peptide and therefore allows it to ‘dissolve’ in venom. However, the tetramer dissociates at or near cell membranes into which monomeric mellitin embeds itself.

Mellitin

Mellitin

And this is where the pain and damage start …

Membrane-association causes cell lysis 3. This results in the release of all sorts of cytokines from the cells which signal ‘damage’ to the body, leading to the inflammatory response usually associated with bee stings. That’s the long-term effect of a bee sting. However, simultaneously, mellitin triggers the expression of proteins known as sodium channels in pain receptor cells. These allow large amounts of sodium to flow across the membrane. It is this that is directly responsible for the pain sensation when you are stung.

So, if being stung is almost inevitable and if bees have evolved stings to cause pain (which they have), in which parts of the body is the pain sensation most marked?

Measuring pain

Pain is a subjective response. What’s painful to me might hardly be noticed by someone with a higher pain threshold. Two individuals receiving the same sensory input can experience very different sensory responses 4.

As an aside it’s well documented that there are differences in the pain felt by males and females 5. All the pain reported in this article is from studies – or personal experience – by males.

Therefore, to meaningfully determine how much pain a sting causes, from a particular insect or at a particular location for example, it’s essential that the studies are properly controlled. This includes taking account of variation between individuals and variation within an individual on a day to day basis.

These are not the sorts of studies that attract large numbers of volunteers 😉

Perhaps unsurprisingly, the scientific work in this field is often published in single author papers in which the author alone is the ‘volunteer’.

The Schmidt Sting Pain Index

Before discussing honey bees specifically a brief diversion must be made to introduce the seminal studies by Justin Schmidt.

Schmidt is an entomologist at the Carl Hayden Bee Research Centre in Arizona. He’s interested in haemolysis (the cell lysis caused by mellitin and other constituents of insect venom) and whether the evolution of sociality in hymenopterans (bees, ant and wasps) required the evolution of toxic and painful stings.

Over about twenty years Justin Schmidt published a number of papers on hymenopteran venoms and the pain that they cause. In his early papers he rated stings on a scale of 1 – 4 (actually 0 upwards, but 0 was totally painless to humans).

Only a very few insects scored 4, including bullet ants about which Schmidt comments “Paraponera clavata stings induced immediate, excruciating pain and numbness to pencil-point pressure, as well as trembling in the form of a totally uncontrollable urge to shake the affected part“.

You can’t fault his commitment, but you might question his sanity.

Schmidt published his magnum opus in 1990 in which he ranked stings by 78 hymenopteran species covering 41 genera 6. His descriptions of the pain induced are often entertaining.  The aforementioned bullet ant is “pure, intense, brilliant pain…like walking over flaming charcoal with a three-inch nail embedded in your heel“.

Another sting scoring 4, that of Synoeca septentrionalis (the warrior wasp) is accompanied by the statement “Torture. You are chained in the flow of an active volcano. Why did I start this list?”.

Why indeed?

Standing on the shoulders of giants

Bees and wasps scored 2 on the Schmidt Sting Pain Index. What Schmidt didn’t investigate was the influence of the location of the sting on the pain experienced.

Which brings me to Michael Smith. In 2014 Smith published an entertaining paper entitled Honey bee sting pain index by body location. It’s published in the journal PeerJ and the full paper is available for download.

It’s a well controlled study written in an engaging style that most readers will appreciate.

Building on the landmark studies by Schmidt, Michael Smith rated the pain endured from honey bee stings in 25 different locations.

Some of these locations should really be protected with a bee suit.

Sting locations tested

Sting locations tested

Smith controlled the study by always including an “internal control” i.e. comparing two test locations with three stings to his forearm.

Every time.

All locations were tested in triplicate (randomly). This meant that Smith was stung a minimum of five times for 38 consecutive days. Ouch.

There are some excellent quotes in the paper … “Some locations required the use of a mirror and an erect posture during stinging (e.g., buttocks)“. Scientists involved in studies that require ethical approval will appreciate the comments made in the paper on self-experimentation.

And the results? To quote directly from the paper “The three least painful locations were the skull, middle toe tip, and upper arm (all scoring a 2.3). The three most painful locations were the nostril, upper lip, and penis shaft (9.0, 8.7, and 7.3, respectively)” 7. Interestingly, skin thickness did not correlate with the pain experienced.

My experience of stings is limited. Those I’ve had to the face (including the lower lip) have been relatively painless, but the subsequent inflammatory response has been dramatic. Smith only scored immediate pain … I think a follow-up study on inflammation and its duration is needed.

I’m not going to conduct it.

Points pain means prizes

You can’t fault the dedication shown by Justin Schmidt and Michael Smith 8. That sort of dedication should be recognised with prizes and honours.

And it was.

Schmidt and Smith shared the 2015 Ig Nobel prize for Physiology and Entomology. The Ig Nobels – a parody of the Nobel prizes – recognise unusual or trivial achievements in scientific research.

The list of Ig Nobel prizes awarded is eclectic and highly entertaining … Medicine (2013) “for treating “uncontrollable” nosebleeds, using the method of nasal-packing-with-strips-of-cured-pork“, Economics (2005) for the inventors of “Clocky, an alarm clock that runs away and hides, repeatedly, thus ensuring that people get out of bed, and thus theoretically adding many productive hours to the workday” and Psychology (1995) for “their success in training pigeons to discriminate between the paintings of Picasso and those of Monet9.

Sir Andre Geim received the Physics Ig Nobel in 2000 for levitating a frog by magnetism. Yes, really. Ten years later he was awarded the Nobel prize in Physics for his studies on graphene. He’s the only holder of a Nobel and Ig Nobel.

Marc Abrahams, the founder of the Ig Nobel awards, regularly tours giving talks on Improbable Research and the Ig Nobel prizes.

Go if you get the chance … it’s highly entertaining.


 

 

Survival of the fattest

Winter bees have high levels of vitellogenin, a glycolipoprotein 1, deposited in their fat bodies which act as a food reservoir for the long winter.

These fat winter bees are essential for the successful overwintering of the colony.

Last week I discussed the major points that need attention for overwintering i.e. strong, healthy colonies with ample food in a weathertight hive.

This week I want to explore the relationship between colony strength, health – specifically with regard to Varroa and deformed wing virus (DWV) – and isolation starvation.

Isolation starvation describes the phenomenon where a small colony of tightly clustered honey bees gets isolated from the honey stores laid down in autumn, resulting – typically during protracted cold periods – in the colony starving to death.

Isolation starvation ...

Isolation starvation …

It’s both a pathetic and distressing sight. Bees, with their heads crammed into the bottom of cells searching for food, dying from starvation when literally inches away from capped stores.

Deaths and births

In temperate climates the winter is characterised by low temperatures and little or no forage for the bees. The queen usually stops laying sometime in autumn and starts again around the turn of the year. During the intervening period she may lay intermittently, but generally in limited amounts.

The fat bodied winter bees that are reared in late summer and early autumn are long-lived (about 6 months) and are responsible for getting the colony through the winter. They protect the queen, thermoregulate the hive and they help rear the brood raised in the autumn and through the winter.

In their absence – or if there are just too few of them – the colony will perish.

Winter bees do not all live for 6 months. The usual figure quoted is ~175 days 2. Some live shorter lives, some longer … up to 9 months under certain conditions.

Importantly, in studies I’ve discussed at length previously, high levels of DWV reduces the lifespan of winter bees. We know this because, in Varroa-infested colonies, researchers 3 have shown that the winter bees die off faster 4.

Live fast, die young

Winter bees with high levels of DWV don’t really live fast … but they do die young. In the studies above the average lifespan of winter bees was reduced by 20% in the colonies that died overwinter.

There are a couple of important things to note here. Dainat and colleagues were not looking at bees in the presence or absence of Varroa, or in the presence or absence of high or low levels of DWV. They simply looked at hives that succumbed in the winter or that survived, then measured DWV and Varroa levels. It’s a subtle but important difference. Their surviving colonies still had Varroa and DWV.

From analysis of hives that died or survived, and having marked known numbers of bees in late summer, they could determine the life expectancy of workers – in their surviving colonies it was ~88 days, in those that died it was ~71 days.

Healthy colonies

The gradual death of bees through the winter coupled with the reduced lifespan of winter bees with high levels of DWV explains why colonies need to be strong and healthy.

The following graphs are based upon modelled data 5, but show the influence of colony size and winter bee lifespan.

The first graph – the least important – simply shows the lifespan of bees. The graph plots the number of bees (on the vertical axis) in a population that die at a particular time (on the horizontal axis) after the start of the experiment. The blue bees have a longer average lifespan than the red bees 6.

Lifespan of winter bees

Lifespan of winter bees

In the following graphs remember that the blue bees are healthy, with low levels of Varroa and – consequently – low levels of DWV. The red bees are unhealthy and have high levels of Varroa and DWV.

Using this lifespan data we can look at the influence on the total number of winter bees in a colony (on the vertical axis) over time (horizontal). Imagine that the horizontal axis is the long, dark, wet and cold months of winter. Starting in early September and running through until late March.

Brrrr 🙁

Winter bee numbers in healthy (blue) and unhealthy (red) colonies

Winter bee numbers in healthy (blue) and unhealthy (red) colonies

It is clear, and of course entirely predictable, that the numbers of bees in the healthy (blue) colony are higher than those in the unhealthy colony at each time point. If the average lifespan is reduced (by disease) more bees will have died by a particular time point when compared with a healthy colony at the same timepoint.

Finally, consider that the shaded section of the graph represents the lower limit of bee numbers for viability. If the number of bees in the colony drops into this region the colony will perish.

Simplistically – and in reality – starting with similar numbers of bees a healthy colony will survive longer than an unhealthy colony.

Strong colonies

Using a similar approach we can also look at the influence of the average lifespan of winter bees on the survival of strong or weak colonies.

The following graph shows the numbers of bees in the colony over time for a strong colony (solid line) and a weak colony (dashed line) where worker bee lifespan is identical 7.

Winter bee numbers in strong and weak colonies.

Winter bee numbers in large (strong) and small (weak) colonies with the same average lifespan.

The shaded section of the graph again represents colony oblivion.

Large (strong) colonies take longer to drop below the threshold for viability and so – all other things being equal – will survive longer 8.

Mix’n’match

A strong colony with high levels of Varroa and DWV might actually survive less well than a weak but healthy colony.

Strong unhealthy colonies might survive less well than weak healthy colonies.

Large unhealthy colonies might survive less well than small healthy colonies.

In this graph the weak but healthy colony drops below the ‘viability threshold’ after the strong but unhealthy colony 9.

Winter bees and brood rearing

This is modelled data, but it makes the point clearly. Large and/or healthy colonies retain more of the all-important winter bees and so survive longer.

Simples.

The differences might not appear marked. However, for convenience 10 I’ve omitted the influence of winter bee numbers on the ability of the colony to rear brood.

If there are more winter bees, the colony is able to thermoregulate the hive better. It’s therefore able to keep any brood present warm. It’s therefore able to rear more brood.

As a consequence, the differences in bee numbers between the large or small, or the healthy and unhealthy, colonies will be much more striking.

Critically 11 the strength of the colony coming out of the winter is often the rate-limiting determinant for spring build-up to exploit early season nectar flows. Weak colonies develop less well.

Isolation starvation

Finally, returning to that pathetic little cluster of starving bees in the image at the top of the page. What is the relationship between colony health, strength and isolation starvation?

It’s now time to dust off my weak-to-non-existent Powerpoint skills …

Isolation starvation schematic

Isolation starvation schematic

Again, it’s straightforward. A large (strong) overwintering colony (A above) only has to move a short distance to access stores in midwinter. In contrast, a small (weak) overwintering colony has to move much further.

Consequently, small colonies become isolated from their stores during long, cold periods when the colony is clustered.

Prediction

Many beekeepers will be familiar with isolation starvation of overwintering colonies.

Most would explain this in terms of “very cold weather and the cluster was unable to reach its stores”.

Some would explain this in terms of “the colony was far too small to reach the stores when clustered”.

Very few would explain this in terms of “the Varroa and DWV levels were too high because of poor disease management last autumn. Inevitably most of my winter bees died off early in the winter, leaving a very small cluster of bees that were unable to reach the stores..

I suspect the real cause of isolation starvation is probably disease … specifically poor management of Varroa levels and consequently high levels of DWV in the colony.


Colophon

Herbert Spencer

Herbert Spencer

Another post, another poor pun in the title. Survival of the fittest encapsulates the Darwinian evolutionary principle that the form of an organism that survives is the one able to leave the most copies of itself in future generations. Darwin didn’t actually use the term until the 5th edition (1869) of his book On the origin of the species. Instead, the phrase was first used by Herbert Spencer in 1864 after reading Darwin’s book. Whilst ‘survival of the fittest’ suggests natural selection, Spencer was also a proponent of the inheritance of acquired characteristics, Lamarckism.

The day job

It’s no secret that I have both amateur and professional interests in bees, bee health and beekeeping.

During the weekend I sweat profusely in my beesuit, rushing between my apiaries in Central and Eastern Fife, checking my colonies – about 15 at the autumn census this year – averting swarms, setting up bait hives, queen rearing and carrying bulging supers back for extraction.

Actually, not so much of the latter in 2017  🙁  I did get very wet though, much like all the other beekeepers in Fife.

The BSRC labs

The BSRC labs …

During the week I sit in front of a large computer screen running (or sometimes running to keep up with) a team of researchers studying the biology of viruses in the Biomedical Sciences Research Complex (BSRC) at the University of St. Andrews. Some of these researchers work on the biology and control of honey bee viruses.

During the winter the beekeeping stops, but the research continues unabated. The apiary visits are replaced with trips in the evenings and weekends to beekeeping associations and conventions to talk about our research … or sometimes to talk about beekeeping.

Or both.

This weekend I’m delighted to be speaking at the South Devon Beekeepers Convention in Totnes on the science that underpins rational and practical Varroa control.

Which came first?

I’ve been a virologist my entire academic career, but I’ve only worked on honey bee viruses for about 6 years. I’ve been a beekeeper for about a decade, so the beekeeping preceded working on the viruses of bees.

However, the two are inextricably entwined. Having a reasonable amount of beekeeping experience provides a unique insight into the problems and practicalities of controlling the virus diseases that bees get.

Being able to “talk beekeeping” with beekeepers has been very useful – both for the communication of our results to a wider audience and in influencing the way we approach our research.

Increasingly, the latter is important. Researchers need to address relevant questions, using their detailed understanding of the science to deliver practical solutions to problems1. There’s no point in coming up with a solution if there’s no way it’s implementation is compatible with beekeeping.

Deformed wing virus

DWV symptoms

DWV symptoms

The most important virus for most beekeepers in most years is deformed wing virus (DWV). This virus “does what it says on the tin” because, at high levels, it causes developmental defects in pupae that emerge with shrivelled, stunted wings. There are additional developmental defects which are slightly less obvious, but there are additional (largely invisible) changes which are of greater importance.

DWV reduces the lifespan of worker bees. This is probably not hugely significant in workers destined to live only a few weeks in midsummer. However, the winter bees that get the colony through from September through to March must live for months, not weeks. If these bees are heavily infected with DWV they die at a faster rate. Consequently, the colony dwindles and dies out in midwinter or early Spring. At best, it staggers through to March and then never builds up properly. It’s still effectively a winter loss.

Our research focuses on how Varroa influences the virus population. There’s very good evidence now that DWV transmission by Varroa leads to a significant increase in the amount of virus, and a considerable decrease in the diversity of the virus population.

So what?

Well, this is important because if we want to control the virus (i.e. to reduce DWV-associated disease and colony losses) it must help to know the proper identity of the virus we are trying to control. It will also help us measure how well our control works. We know we’re measuring the right thing.

We’re working with researchers around the world to define the important characteristics of DWV strains that cause disease and, closer to home, with entire beekeeping associations to investigate practical strategies to improve colony health.

Chronic bee paralysis virus

CBPV symptoms

CBPV symptoms

We’re about to start a large collaborative project on the biology and control of chronic bee paralysis virus (CBPV). This virus is becoming a significant problem for many beekeepers and is increasing globally. It’s a particular problem for some bee farmers.

CBPV causes characteristic symptoms of dark, hairless, oily-looking bees that sometimes shiver, dying in large smelly piles at the hive entrance. It typically affects very strong colonies in the middle of the season. It can be devastating. Hives that should be the most productive ones in the apiary fail catastrophically.

Why is a virus we’ve known about for decades apparently increasing in the amount of disease it causes? Are there new virulent strains of the virus circulating? Are there particular beekeeping practices that facilitate it’s spread? We’re working with collaborators in the University of Newcastle to try and address these and related questions.

I’ll write more about CBPV over the next year or so. It won’t be a running dialogue on the research (which would be crushingly dull for most readers), but will provide some background information on what is a really fascinating virus.

At least to a virologist 😉

And perhaps to beekeepers.

Grow your own

As virologists, we approach the disease by studying the virus. Although we maintain an excellent research apiary, we don’t do many experiments in ‘the field’. Almost all the work is done in test tubes in incubators in the laboratory … or in bees we rear in those incubators.

Grow your own

Grow your own …

We can harvest day-old larvae (or even eggs) from a colony and rear them to emergence as adult bees in small plastic dishes in the laboratory. We use an artificial diet of sugar and pollen to do this. It’s time consuming – they need very regular feeding – but it provides a tightly controllable environment in which to do experiments.

Since we can rear the bees, we can therefore easily test the ability of viruses to replicate in the bees. Do all strains of the virus replicate equally well? Do some strains outcompete others? Does the route by which the virus is acquired influence the location(s) in the bee in which the virus replicates? Or the strains it is susceptible to? Or the level of virus that accumulates?

And if our competitors are reading this, the answer to most of those questions is ‘yes’ 😉

We can even ask questions about why and how DWV causes deformed wings.

Again, so what? We suspect that DWV causes deformed wings because it stops the expression of a gene in the bee that’s needed to make ‘good’ wings. If we can identify that gene we might be able to investigate different strains of honey bee for variation in the gene that would render them less susceptible to being ‘turned off’ by DWV. That might be the basis for a selective breeding project.

It’s a simplistic explanation, but it’s this type of molecular interaction that explains susceptibility to a wide range of human, animal and plant diseases.

Bee observant

Bee health is important, and not fundamentally difficult to achieve. There are some basics to attend to … strong hives, good forage, good apiary hygiene etc. However, it primarily requires good powers of observation – does something look odd? Are there lots of mites present? How does the brood look?

If things aren’t right – and often deducing this means comparisons must be made between hives – then many interventions are relatively straightforward.

Not long for this world ...

Not long for this world …

The most widespread problems (though, interestingly, this doesn’t apply to CBPV) are due to high levels of Varroa infestation. There are effective and relatively inexpensive ways to treat these … if they’re used properly.

More correctly, they’re relatively inexpensive whether they’re used properly or not. However, they’re pretty ineffective if not used properly 😉

Regular checks, good record keeping, comparisons between hives and informed observation are what is needed. Don’t just look, instead look for specific things. Can you see bees with overt symptoms of DWV? Are there bees with Varroa riding around on their backs? The photo above has both of these in plain view. Are some hairless bees staggering around the top bars with glossy abdomens, or clinging to the side bars shaking and twitching?

Don’t wait, act

I’ve no doubt that scientists will be able to develop novel treatments to control or prevent virus infections of bees. I would say that … I’m a scientist 😉  However, I’m not sure beekeepers will be able to afford them, or perhaps even want to use them, or that they’d be compatible with honey production or of any use in Warré hives etc.

I’m also not sure how soon these sorts of treatments might become available … so don’t wait.

If there are signs of obvious DWV infection you need to do something. ‘Obvious’ because DWV is always present, but it’s usually harmless or at least tolerated by the bees. My lab have looked at thousands of bees and have yet to find one without detectable levels of DWV. However, healthy bees have only about 1/10,000 the level of DWV present in sick bees … and these are the ones that have obvious symptoms.

I’ve discussed Varroa control elsewhere, and will again.

Unfortunately, if your colony has signs of CBPV disease then Varroa control is not really relevant. The virus is transmitted from bee to bee by direct contact. This probably accounts for the appearance of the disease primarily in very strong colonies.

At the moment there’s little you can do to ‘cure’ a CBPV-afflicted colony. I hope, in 2-3 years we will have a better idea on what interventions might work. We have lots of ideas, but there are a lot of basic questions to be addressed before we can test them.

Field work

Field work

Business and pleasure

The half of my lab that don’t work on bee viruses study fundamental mechanisms of virus replication and evolution. They do this using human viruses, some of which are distant relatives of DWV. They work on human viruses as it’s only these that have excellent model systems to facilitate the types of elegant experiments we try to do. They’re also relatively easy to justify in funding applications, and it allows us to tap into a much bigger pot for funding opportunities (human health R&D costs probably total £2 billion/annum, bees might be £2 million/annum).

And no, my lab don’t get anything like that much per year for our research!

Importantly, the two activities on human and honey bee viruses are related. Our experience with the human viruses related to DWV made us well-qualified to tackle the bee virus. They replicate and evolve in very similar ways, we quantify them in the same way and there may be similarities in some ways we could approach to control them.

And with the bee viruses I can mix business with pleasure. If I’m going to the apiary I’ll get to see and handle bees, despite it being officially “work”. It doesn’t happen as much as I’d like as I’m usually sat behind the computer and all of the ‘bee team’ have been trained to work with bees by the ESBA.

However, at least when I talk to collaborators or to the beekeeping groups we’re fortunate to be working with we – inevitably – talk about bees.

And that’s fun  😀


Several years ago I delivered an enthusiastic and rather science-heavy talk at a Bee Farmers Association meeting. I thought it had gone reasonably well and they were kind enough to say some nice things to me … and then I got the question from the back of the room which went something like “That’s all very well young man … but what have you made NOW that I can put into my hives to make them healthy?”.

I’m sure my answer was a bit woolly. These days the presentation would have had a bit less science and bit more justification. We’ve also made some progress and it’s possible to now discuss practical strategies to rationally control viruses in the hive. It’s not rocket science … though some of the science it’s based on is reasonably fancy.

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