Category Archives: Science

Biological control with Varroa

Synopsis : Honey bees were eradicated on Santa Cruz Island following the introduction of Varroa. This provides some useful lessons for beekeepers on the importance of controlling Varroa.

Introduction

Honey bees are not native to North America. They were first introduced in March 1622 at Jamestown, Virginia. The bees did well and spread west, following the settlers. They finally arrived on the west coast, in Santa Clara, California, 231 years later in 1853. Of a dozen hives ordered by Christopher Shelton, a Santa Clara botanist and rancher, only one survived the journey from New York via Panama.

Shelton barely had a chance to enjoy his bees 1 as he was unfortunately killed when the steamboat Jenny Lind exploded in mid-April 1853.

Explosion on the steamboat Jenny Lind near San Francisco, California

His bees survived 2 and three hives derived from the original stock were auctioned for $110 each. This was over 20 times the price of hives on the east coast at that time and equivalent to over $4200 today 3.

Californian Channel Islands map

Bees were in demand and they continued to spread – both as feral swarms and as farmers established apiaries to help pollination and for honey production. Having reached the California coast they were then spread to the nearby islands. Bees were transported to Santa Cruz, the largest of the eight Channel Islands near Los Angeles, in the 1880’s. They flourished, but did not spread to the other Channel Islands.

Field station, nature reserves, pigs and bees

Santa Cruz Island is 250 square kilometres in area and lies ~35 km south of Santa Barbara. It is one of the four Northern Channel islands. There is a long central valley lying approximately east-west and the rocky mountainous land reaches 740 m. It has a marine temperate climate; the average low and high temperatures are 9°C and 21°C respectively and it receives about 0.5 m of rain a year. It is a good environment for bees.

From the 1880’s to 1960’s Santa Cruz Island was farmed – primarily for wine and wool, and from the 1940’s for cattle – but, after period of university geology field trips and the establishment of a field station on the island, in 1973 it became part of the University of California’s Natural Reserve System (UC NRS).

In the late 1970’s the Stanton family sold their ranching business on the island to The Nature Conservancy who subsequently bought additional land on the eastern end of the island.

Santa Cruz Island is now jointly owned by The Nature Conservancy, National Parks Service, UC NRS and the Santa Cruz Island Foundation and much of the island is used for scientific research and education.

But what about the bees?

Good question.

As a nature reserve and research station, the presence of non-native species causes a potential problem. Why go to all the expense of managing a remote island research centre if all the same species are present as on the mainland?

The Nature Conservancy therefore initiated a programme of eradicating non-native species. It took 14 months to eliminate the feral pigs, using a combination of trapping, helicopter-based shooting and the release of sterilised radio-tagged pigs to locate the stragglers 4.

But getting rid of the bees took a bit longer …

Save the bees, or not

Why get rid of the bees? Surely they weren’t doing any harm?

The introduction of any non-native species upsets the balance (if there’s ever balance) in the ecosystem. The introduced species competes directly or indirectly with those native to the area and can lead to local extinctions.

Jonathan Rosen has described 5 how honey bee swarms, through occupying tree cavities previously used for nesting, probably played a major role in the extinction of the Carolina parakeet.

Pining for the fjords … a stuffed Carolina parakeet (nailed to its perch)

Competition between honey bees and native pollinators has been well studied. It is not always detrimental, but it certainly can be. Furthermore, it is probably more likely to be detrimental in a small, isolated, island ecosystem. For example, studies showed that the presence of honey bees dramatically reduced visitation of native pollinator to manzanita blossoms on Santa Cruz Island.

As part of the larger programme of non-native plant and animal eradication on Santa Cruz Island plans were drawn up in the late 1980’s to eliminate European honey bees. The expected benefits were to:

  • eliminate competition with native bee species (and presumably other non-bee pollinators, though these rarely get a mention 🙁 )
  • reduce pollination of weed species (some of which were also non-native to Santa Cruz Island)
  • facilitate recovery of native plant species that were reliant on native bee pollination
  • provide a ‘field laboratory’ free from ‘exotic’ honey bees in which comparative studies of native pollinators would be possible

Killer bees

After the plans to eradicate Apis mellifera were approved an additional potential benefit became apparent.

There were increasing concerns about the spread of Africanised honey bees which had recently reached Santa Barbara County. Although there was reasonably compelling evidence that swarms could not cross from the mainland (e.g. none of the other Northern Channel Islands had been colonised by bees) there were concerns that the Santa Ana winds might help blow drones from the mainland.

Had these drones arrived they might mate with the non-native but nevertheless local queens resulting in the spread of the dominant genes for defensiveness and absconding. The resulting swarmy, aggressive Africanised bees would cause problems for visitors and scientists working on the island (as they have for visitors to Joshua Tree National Park).

Aerial view of Santa Cruz Island

Although the introgression of African honey bee genes was used as further justification for the eradication it’s not clear whether drones could actually cross 30-40 km of open sea 6.

As an aside, there’s a current project – the amusingly named Game of Drones – running on the Isles of Scilly investigating whether drones can cross the sea between St Agnes, Tresco, Bryher, St Mary’s and St Martin’s. These are, at most, 11 km apart (northern most tip of St Martin’s to most southerly point of St Agnes) but the individual islands are only separated by 1-2 km. I would be surprised if drones could not cross that distance (at least with a strong following wind).

Killing bees

Adrian Wenner and colleagues set about exterminating the honey bees on Santa Cruz Island (Wenner et al., 2009). The process started in 1988 and ended in 2007, and was divided into four phases:

  1. 1988-1993 – location and elimination of feral colonies
  2. 1994-1997 – biological control and colony demise
  3. 1998-2004 – monitoring residual honey bee activity
  4. 2005-2007 – confirmation of the absence of honey bees

None of this is ’beekeeping’ – actually it’s the exact opposite – so I don’t intend to dwell in much detail on the work that was conducted. However, the ’94-’97 phase includes some sobering lessons for beekeepers which are worth discussing.

By the end of phase 1 the team had identified the existence (if not the location) of at least 200 colonies and eliminated 153 of them.

Remember, none of these were managed colonies in hives. They were all feral colonies occupying natural cavities in trees or rocks etc. Each colony was found using painstaking bee lining techniques similar to those described in Thomas Seeley’s book Following the Wild Bees.

Once located, nests were destroyed with methyl chloroform and the cavity sealed to prevent it being reoccupied.

Some colonies could not be accessed; in these cases acephate-laced sucrose-honey syrup baits were used. This organophosphate has delayed toxicity for bees, allowing foragers to return to the colony which in due course dies. This approach had been partially successful in eliminating Africanised bees on the mainland (Williams et al., 1989), but baits needed to be be monitored to avoid killing the other insects they attracted.

The scientists also deployed swarm traps (aka bait hives) and destroyed any swarms that moved in.

Together these interventions reduced honey bee numbers significantly – as monitored by regular observations at pollen- or nectar-rich plants – but did not eradicate them.

Let there be mite

Heavy rains in January ’93 washed out roads on Santa Cruz Island, thereby severely limiting travel around the island. In addition, the previous removal of cattle had resulted in the near-uncontrolled growth of fennel which now formed dense, impenetrable thickets.

Bee lining became impossible and the scientists had to invent more devious strategies to eliminate the residual feral colonies.

The approach they chose involved the introduction of Varroa.

Varroa was first detected in the USA in 1987 (in Florida) and became widespread over the next 5-8 years. Up until 1994 the honey bees on Santa Cruz Island were free of the ectoparasitic mite.

It was likely that they would have remained that way … there was no beekeeping on Santa Cruz Island and the location was too remote for bees to cross from the mainland (see above).

Varroa was already known to have a devastating impact on the health of honey bee colonies (Kraus and Page, 1995). It was also known that, other than its native host Apis cerana (the Eastern honey bee), Varroa did not parasitise other bee or wasp species (Kevan et al., 1991).

These two facts – host specificity and damage inflicted – suggested that Varroa could be used for biological control (‘biocontrol’) on Santa Cruz Island.

Biological control

Biological control or biocontrol is a method of controlling pests using natural mechanisms such as predation or parasitism.

The pest could be any living thing – from animals to bacterial plant diseases – present where it’s unwanted.

On Santa Cruz Island the pest was the honey bee.

In other studies (covered in a previous post entitled More from the fungi 7 ) biocontrol of Varroa has been investigated.

Control of the pest involves the introduction or application of a biological control agent. The key requirements of the latter have already been highlighted – specificity and damage.

Biological control works well when the specificity is high and the damage is therefore tightly targeted. It can be an abject failure – or worse, it can damage the ecosystem – if the specificity is low and/or the damage is widespread.

The cane toad was introduced to Australia to control infestations of greenback cane beetle (a pest of sugar cane). Cane toads were introduced in 1935 and rapidly spread. Unfortunately, cane toads can’t jump very high and so singularly failed to control the greenback cane beetle which tends to 8 stay high up the cane stems.

Female cane toad (not jumping)

But it gets worse; cane toads have a very catholic diet and so outcompeted other amphibians. They introduced foreign diseases to the native frogs and toads and – because of the poisons secreted from their skin – harmed or killed predators that attempted to eat them.

Oops.

Vertebrates are usually poor biological control agents as they tend to be generalist feeders i.e. no specificity.

But Varroa is specific and so the damage it causes is focused. The likelihood of ecosystem damage was considered low and so the mite was introduced to the island.

Introduction of Varroa

In late 1993 Adrian Wenner caught 85 foraging bees and, to each one, added a single Varroa mite. The bees were then released and presumably flew back to their colonies … taking the hitchhiking mite with them.

Adult mites – the dark red ones you see littering the Varroa tray after you treat with Apivar – are mated females.

Due to their incestuous lifestyle a single mite is sufficient to initiate a new infestation.

The mated adult female mite parasitises a honey bee pupa and produces a series of young; the first is male, the remainder are female. You’re probably reading this before the 9 pm watershed so I’ll leave it to your lurid imagination to work out what happens next (or you can read all the sordid details in Know your enemy).

The presence of honey bees – determined by successful swarm trapping or field observation at likely sites – was then regularly monitored over the next four years.

Swarm numbers remained largely unchanged until 1996 and then dramatically decreased.

Numbers of new swarms on Santa Cruz Island 1991 – 2005. Varroa introduction indicated.

It’s worth noting that during ’94-’96 over 70 swarms were found in natural sites or bait hives. There must have been a significant number of established colonies in 1993 to produce this number of swarms.

But, from 1997 it all stopped … only a single swarm was subsequently found, in a natural cavity in 2002.

Monitoring and confirmation of eradication

From 1998 to 2004 the scientists continued to actively monitor the island for honey bees, focusing on 19 areas rich in natural forage. Although honey bees were found – in decreasing numbers – there were too few to attempt bee lining to locate their colonies.

At the sites being monitored, bees were detected 9, 7, 4, 2 and 1 times respectively in the 5 years from 2000 to 2004. After that, despite continued monitoring, no more honey bees were detected.

The final phase of the project (’05-’07) confirmed the absence of honey bees on Santa Cruz Island.

Whilst, as a scientist, I’m a firm believer that ’absence of evidence does not mean evidence of absence’, as a beekeeper I’m well aware that if there are no scout bees, no swarms and no foragers (when I search in likely places) then there are no honey bee colonies.

Lessons for beekeepers

I wouldn’t have recounted this sorry tale – at least from a beekeeping perspective – unless I thought there were some useful lessons for beekeepers.

There are (at least) three.

The first relates to Varroa resistance, the second to Varroa transmission in the environment and the last to ‘safe’ levels of Varroa. All require some ‘arm waving guesstimates’ 9, but have a good grounding in other scientific studies.

Varroa resistance

There wasn’t any.

At a very conservative estimate there were at least 20 colonies remaining on Santa Cruz Island in 1995. I say ‘conservative’ because that assumes each colony generated two swarms that season (see graph above). In studies of other natural colonies only about 75% swarm annually, meaning the actual number of colonies could have been over 50.

The numbers – 20 or 50 – matter as they’re both much higher than the number of colonies most beekeepers manage (which, based upon BBKA quoted statistics, is about 5).

Whether it was 20 or 50, they were all eliminated following the introduction of 85 mites. Colonies did not become resistant to Varroa.

This all took a few years, but – inferring from the swarm numbers above – the vast majority of colonies were killed in just two years, 1994 and 1995. This timing would fit with numerous other studies of colony demise due to mites.

Wenner estimates that only 3 colonies survived until 2001.

Leaving small numbers of colonies 10 untreated with an expectation that resistance – or even tolerance (which is both more likely and not necessarily beneficial) – will arise is a futile exercise.

I’ve discussed this before … it’s a numbers game, and a handful of colonies isn’t enough.

Varroa spread

Wenner doesn’t elaborate on where the foragers were captured before he added the mites. If I was going to attempt this I’d have chosen several sites around the island to ensure as many feral colonies as possible acquired mites … let us assume that’s what he did.

However, with 85 mites piggybacking on returning workers, and somewhere between (my guesstimated) 20 to 50 colonies, I think it’s highly likely that at least some colonies received none of this ’founding’ mite population.

Yet almost all the colonies died within two years, and those that did not subsequently died with no further intervention from the scientists. We don’t know what killed off the last surviving colonies but — and I know I’m sticking my neck out here – I bet it was the mites.

This is compelling evidence for the spread of Varroa throughout the island environment, a process that occurs due to the activities of drifting and robbing.

If a neighbouring apiary to yours has mites some will end up in your hives … unless you are separated by several kilometres 11.

The transmission of mites in the environment is a very good reason to practice coordinated Varroa control.

One mite is all it takes

But, just as I’ve argued that some colonies may have received none of the founding mites, I’m equally sure that others will have acquired very small numbers of mites, perhaps just one.

And one mite is all it takes.

Without exceptional beekeeping skills, resistance in the bee population or rational Varroa control 12 there is no safe level of mites in a colony.

The more you prevent mites entering the colony in the first place, and the more of those that are present you eradicate, the better it is for your bees.

Here endeth the lesson 😉


Note

It’s worth noting that island populations do offer opportunities for the development of Varroa resistant (or tolerant) traits … if you start with enough colonies. Fries et al., (2006) describes the characteristics of the 13 surviving colonies on Gotland after leaving about 180 colonies untreated for several years. I’ve mentioned this previously and will return to it again to cover some related recent studies.

References

Fries, I., Imdorf, A. and Rosenkranz, P. (2006) ‘Survival of mite infested (Varroa destructor) honey bee (Apis mellifera) colonies in a Nordic climate’, Apidologie, 37(5), pp. 564–570. Available at: https://doi.org/10.1051/apido:2006031.

Kevan, P.G., Laverty, T.M. and Denmark, H.A. (1990) ‘Association of Varroa Jacobsoni with Organisms other than Honeybees and Implications for its Dispersal’, Bee World, 71(3), pp. 119–121. Available at: https://doi.org/10.1080/0005772X.1990.11099048.

Kraus, B. and Page, R.E. (1995) ‘Effect of Varroa jacobsoni (Mesostigmata: Varroidae) on feral Apis mellifera (Hymenoptera: Apidae) in California’, Environmental Entomology, 24(6), pp. 1473–1480. Available at: https://doi.org/10.1093/ee/24.6.1473.

Wenner, A.M., Thorp, R.W., and Barthell, J.F. (2009) ‘Biological control and eradication of feral honey bee colonies on Santa Cruz Island, California: A summary’, Proceedings of the 7th California Islands Symposium, pp. 327–335. Available as a PDF.

Williams, J.L., Danka, R.G. and Rinderer, T.E. (1989) ‘Baiting system for selective abatement of undesirable honey bees’, Apidologie, 20(2), pp. 175–179. Available at: https://doi.org/10.1051/apido:19890208.

 

Making a beeline

Synopsis : Honey bees use a range of navigation skills including path integration – to shorten return flights – combined with map-like spatial memories to relocate the hive.

Introduction

Regular readers will be aware that I’m interested in the origins of words. The Oxford English Dictionary (OED) is a fantastic source of information and produces a free Word of the Day email 1. This includes both the meaning and etymology of one word each day.

Since the complete dictionary includes over 600,000 words it will take a few years to collate the 20 volumes that comprise the entire dictionary 2.

At the beginning of this week the word of the day was beeline.

The word beeline of course means:

A straight line or course, such as a bee follows in returning to its hive after having collected a full load of nectar; (occasionally) the course taken by a bee.

The word originated in the US almost 200 years ago. It was first recorded in the American Quarterly Review in June 1828. Anyone who has read Tom Seeley’s Following the Wild Bees will appreciate the context in which the word beeline was used:

The bee-hunter..encloses them [sc. bees] in a tube, and letting one fly, marks its course, by a pocket compass. Departing to some distance, at right angles to the bee-line just ascertained, he liberates another, observes its course, and thus determines the position of the hive, which lies in the angle made by the intersection of the bee-lines.

Beelining is the art of finding feral or wild colonies by following the returning flight of bees. The book has a companion website with some interesting videos if you’d like to know more.

Find and tell

Beelining ‘works’ because bees fly in a straight line back to the nest 3.

The basics of beelining

Assume the blue flowers above are nectar-rich and favoured by the bees. You capture a couple of bees feeding on the blue flowers and give them some additional syrup so that they are replete and need to return to the colony to unload.

When you release the bees at ‘A’ they fly at a particular bearing back to the colony. However, if you instead release them at ‘B’ they fly at a different compass bearing back to the colony.4  .

How did the bees find the nectar-rich blue flowers in the first place?

Perhaps they observed another worker in the colony performing the waggle dance which informed them of the angle (from the sun) and distance to the blue flowers?

Alternatively, they might have just been searching around and chanced upon the blue flowers … they didn’t know they were there in the first place.

If they found the blue flowers by interpreting the waggle dance then you should be thinking how the waggle dancing bee found the blue flowers.

Alternatively, if they found the blue flowers by chance then you should be wondering how they will communicate their location to other foragers in the colony.

Transient nectar sources

Nectar sources are transient. They yield at particular times of the year … and of the day. The nectar may be dependent on recent rainfall or a variety of other environmental conditions.

All this means is that foragers may have to search widely to find a good source of nectar. If the source is really good – ample sugar-rich nectar and with lots of flowers producing it – then it’s important that the forager that found it tells her half-sisters how to also quickly find the same source.

Foraging and finding

On the left the blue flowers have been yielding for days. The workers fly there in a straight line and return along the same path. Newly orientated workers observe the returning foragers waggle dancing and follow the same route to quickly and efficiently exploit the source.

But all good things come to an end …

On the right is what happens when blue flowers stop yielding. The foragers that arrive at the blue flowers find slim pickings and start casting about looking for a better source of nectar. They first find the marginally better yellow flowers, then the similar (but far from outstanding) purple flowers … so they keep looking.

And eventually, they find the red flowers. Lots of nectar and lots of flowers. They load up and return directly to the colony (black dotted line).

There are two striking things about this return flight. The first is that it does not follow (in reverse) the route by which they reached the red flowers. The second is that when these returning foragers perform the waggle dance they ‘instruct’ the observing bees to fly in the direction of the red dotted line … rather than to the blue, then yellow, then purple and then red flowers.

Path integration

The foragers who find the red flowers perform a process termed path integration to return:

Path integration is the process by which an animal, when moving away from a start point, often its nest, cumulatively sums its path, generating an internal vector that specifies the line from the animal’s current position back to the start point, however circuitous the outward trip (Collet, 2019).

This is a skill I singularly lack when trying to relocate my vehicle in the multi-storey car park.

Path integration is seen in other insects … Drosophila fruit flies can do it (over a range of centimetres), walking ants can do it over a range of hundreds of metres, and honey bees can do it over at least 5 kilometres (and probably more).

Path integration requires two pieces of information – the direction and the distance of travel.

Path integration – individual parts of the flight are in different directions and of different lengths

Clearly, the very existence of the waggle dance provides compelling evidence that bees are aware of both. The dancing forager reports the angle (relative to the sun) of the nectar source and the distance at that angle that must be covered before the nectar source is located.

But for path integration, not only must the angle and distances be determined, they must also be cumulatively summed.

Neurophysiology and evolutionary conservation

Detailed neurophysiological experiments – recording the firing of individual neurones in the bee’s brain – have identified that these events occur in a region called the central complex (CX).

Two types of neurones are involved; the first is a set of polarised-light-based compass neurones and the second are optic-flow-based speed neurones. The former use celestial cues to create a visual compass. The latter provide a visual odometer (Stone et al., 2017).

Together – and there are additional integrator cells that link these functions – this relatively simple 5 neuronal circuitry allows path integration, enabling the bee to return ‘home’ directly after a convoluted outward flight.

Many of these studies were conducted on the nocturnal sweat bee Megalopta genalis. This forages at night when polarised skylight provides the directional cues in its rainforest habitat.

Importantly, similar neuronal organisation is found in the CX’s of locusts, some butterflies and dung beetles. The visual odometer neurones were analysed in Megalopta genalis, but are physically and likely functionally similar to structures found in Bombus terrestris (a bumble bee).

You may have noticed that none of these studies used our favourite, Apis mellifera, the honey bee.

The evolution of termites, ants, wasps and bees

Nevertheless, there’s every reason to think that honey bee path integration involves very similar neuronal activity. Megalopta (belonging to the family Halictidae) and Bombus (a member of the Apidae family) are very distantly related and evolved from a common ancestor over 100 million years ago (Cardinal and Danforth, 2011). It’s therefore likely that all bees derived from this common ancestor – including honey bees – share similar neuronal activity underpinning their path integration ability.

Food vectors

Before considering another point about honey bee flight I wanted to to briefly mention features of the outbound trip back to the high quality food source (the red dotted line in diagram above). This is termed the food vector and is essentially the reverse of the path integrated return flight back to the colony i.e. the same length, but pointing in the opposite direction.

The waggle dance communicates this food vector to nest mates of the successful returning forager.

But what happens if bees are displaced when starting, or while following this food vector?

For example, if a huge gust of wind blew them off course by tens or hundreds of metres, or an evil eager scientist captured them as they left the hive and transported them in a dark box across a couple of fields and then released them?

Where do displaced foragers go?

Do the bees fly a corrected route to the food source (the blue dotted arrow), or do they continue flying the same vector (angle and distance – the green dotted arrow) they would have done when they left the hive?

I’m not sure this exact experiment has been done with bees (but see below), but it has been done with ants (Cataglyphis fortis). In these studies the ’displaced’ ants did alter their direction of travel (Collett et al., 1999). The food vector is more than just an angle and distance, it also points to a position relative to the nest. The redirection exhibited by the ants was not perfect, but it clearly showed they were able to integrate the path to a location other than the nest after displacement.

Gusts of wind are not the same as eager scientists

However, back to the bees.

The gust of wind and eager scientist are not equivalent. Bees cope with gusts of wind every day. It always amazes me how well bees cope on windy days.

When blown off course they will get lots of visual cues – not least changes in optic flow and their angle to the sun – both of which should be readily corrected. If they didn’t then foragers would be lost in droves on windy days … or fail to find the food source.

In contrast, the eager scientist took care to place the bees in a darkened box, thereby immediately removing visual cues such as the angle of the sun and the optic flow.

In the studies conducted with the ants the scientists made sure the ants could see the sky but not the surrounding landscape (they trained them in open topped channels). This is because ants can also use landmarks in the surrounding landscape for orientation 6.

And bees can do the same, which is the final sub-topic for this post on bee flight and orientation.

The map-like spatial memory of bees

Path integration is both useful and necessary. It means that foragers can return – fully laden – with minimum delay to the hive. They can therefore tell other foragers (via waggle dancing) promptly, and – in the case of elite foragers – they can set off again on another trip.

By reducing the distance flown – by integrating the path – they save not only time but ‘fuel’ as well i.e. path integration allows bees to maximise the nectar returned at the end of the foraging trip.

But, if all flights were a combination of random searches and path-integrated returns, why do bees go on orientation flights?

Orientation flights are short range (10’s to 100’s of metres) flights around the hive. These are taken by workers around 3 weeks after emergence as they transition form hive bees to foragers. They are also taken by older foragers if the hive is moved.

The very existence of orientation flights is compelling evidence that honey bees also use learned environmental landmarks for route finding, or at least for mapping the area around the hive to aid efficient return trips.

What evidence is there that these landmarks are used for this purpose?

Harmonic radar tracking of displaced foragers

I’ve previously discussed the use of short range harmonic radar to track bees ‘tagged’ with a small transponder. The key point is that it allows relatively accurate mapping of the entire flight of a bee up to 900 metres away. The resolution is, at best, about 3 metres.

Menzel and colleagues (Menzel et al., 2004) tracked the flights of three types of ‘displaced’ foragers:

  • SF-bees trained to a stationary feeder a few hundred metres from the hive; these have ‘route memory’ and have traversed the route from the hive to the feeder multiple times
  • VF-bees trained to a regularly moved feeder within 10 metres of the hive; these bees have no route memory
  • R-bees which were recruited by a waggle dancing forager and have only secondhand route information of the position of the feeder i.e. they have never made the trip themselves

These are not trivial experiments. To ensure the environment was as uniform as possible they conducted the experiments in a large, flat mown field approximately 800 metres square. There was no forage within the field other than the experimental feeders. The field was surrounded on all sides by uniform coniferous woodland with insufficient variation in elevation (<1.5°) above the horizontal to provide any visual clues to the bees.

The field itself was not uniform. There were differences due to different mowing times and soil conditions. In addition, the scientists erected a number of radar-transparent coloured tents around the hive to provide additional landmarks.

Common features of flight paths determined by harmonic radar studies

Bees were allowed to orientate to the new hive position and then SF- and VF-bees were collected at a feeder and R-bees were captured as they left the observation hive (having ‘watched’ a waggle dance). The bees were fitted with a transponder, released some distance away from the feeder or the hive and then tracked by radar.

SF- and VF-bees were stuffed full of syrup and so – although they could fly for a long time – were motivated to return to the hive to unload their cargo. R-bees, whilst ‘primed’ to seek the feeder, had limited range and so would have to return to the hive to refuel.

Return flights of SF-, VF- and R-bees show some common features.

The SF- and R-bees exhibited three broadly conserved flight patterns during their return trip to the hive:

  1. A fast (20 m/s) straight line flight in the direction they would have taken back to the hive (for the SF-bees) or out to the feeder (for the R-bees). The length of this part of the flight was approximately the distance between the hive and the feeder.
  2. A slow (13 m/s) curved search flight.
  3. A fast homing flight back to the hive.

The VF-bees only exhibited the slow curved search flight and the final fast homing flight. This was unsurprising as they had never learned (or been told) to follow the route between the hive and distant feeder.

Food vectors and von Frisch

We therefore have the answer to the question I posed earlier (in the Food vector section above). A bee displaced when about to embark for the first time on a trip to a distant feeder – learnt from following a waggle dance – initially flies at the angle and to the approximate distance they would have taken from the hive (stage 1 of the flight).

Remember, unlike the ants, these foragers are ‘in the dark’ while being displaced, so have no visual clues about the displacement.

This is a really nice result and supports the contention made by von Frisch that the waggle dance communicates only distance and direction (relative to the sun) information, rather than positional information (von Frisch, 1967) 7.

Homeward bound

After a period of slow curved flights the returning forager switches to a direct, fast homing flight. These started at positions – starred in the figure above – from which the bee could not see the hive (based upon distance and the known resolution of honey bee vision).

Homing flights of displaced SF-, VF- and R-bees (A, B, C respectively). H indicates the position of the hive.

Individual bees were randomly displaced around the study field. The homing flights were in a straight(ish) line and bees approached the hive from a range of different points of the compass. This argues strongly against the bees following a particular feature on the ground that led them back to the hive.

Instead, the authors argue that, since all the bees exhibit these direct homing flights, it must be based upon previous exploratory memory i.e. from orientation flights.

The tents were not critical landmarks. If they were moved some distance away the bees still returned using the same three flight phases (in the case of SF- and R-bees) and with similar navigational performance. Clearly there was sufficient information in the ground structure alone (mowing patterns, soil differences) acquired during the orientation flights.

In support of this, some of the harmonic radar data showed bees flying along boundaries between mown areas (in a similar way to homing pigeons follow rivers or motorways; Guilford and Biro, 2014.).

These experiments indicate that during orientation flights the bee develops a local spatial memory of landmarks that provide a ‘memory map’. This enables the bee to return to the nest once it recognises some of these familiar landmarks.

Repeated displacement flights of the same bee further indicated that the landmarks recognised (whatever they were) could be approached from different angles.

Final inspections

My bees are still out foraging despite the large blocks of fondant most hives are now topped with. I’m not sure what they’re collecting but it’s clearly worth the trip … and going to the initial trouble of finding it and telling other foragers about it.

Returning foragers

We usually take the amazing navigational abilities of our bees for granted. Those returning foragers are using navigational skills that evolved at least 100 million years ago while dinosaurs roamed the earth.

100 million years is a long time to develop a range of skills and subtleties; it’s no wonder we still only partially understand honey bee navigation. Of course, we don’t have to understand it to still marvel at their ability to find the way back.

And it’s worth also remembering that these navigation skills – many of which are based upon the angle of travel relative to the direction of the sun – also operate on dull, overcast days. But that’s a topic for another post …


References

  • Cardinal, S. and Danforth, B.N. (2011) ‘The Antiquity and Evolutionary History of Social Behavior in Bees’, PLOS ONE, 6(6), p. e21086. Available at: https://doi.org/10.1371/journal.pone.0021086.
  • Collett, M., Collett, T.S. and Wehner, R. (1999) ‘Calibration of vector navigation in desert ants’, Current Biology, 9(18), pp. 1031–1034. Available at: https://doi.org/10.1016/S0960-9822(99)80451-5.
  • Guilford, T. and Biro, D. (2014) ‘Route following and the pigeon’s familiar area map’, Journal of Experimental Biology, 217(2), pp. 169–179. Available at: https://doi.org/10.1242/jeb.092908.
  • Menzel, R. et al. (2005) ‘Honey bees navigate according to a map-like spatial memory’, Proceedings of the National Academy of Sciences, 102(8), pp. 3040–3045. Available at: https://doi.org/10.1073/pnas.0408550102.
  • Stone, T. et al. (2017) ‘An Anatomically Constrained Model for Path Integration in the Bee Brain’, Current Biology, 27(20), pp. 3069-3085.e11. Available at: https://doi.org/10.1016/j.cub.2017.08.052.

More droning on …

Synopsis : Drones are now being evicted from colonies. How and why does a honey bee colony regulate drone numbers?

Introduction

Over the course of the last eight years posts on The Apiarist have got longer. This year, posts are now five times the length of the 2014 average. I’ve written – and hopefully, you’ll have already read – more words this year than are in The Hobbit.

If this continues until the end of the year we’ll have exceeded the word count in Tolkien’s The Two Towers.

This is probably unsustainable 1.

The increase is explained in part by the complexity of some topics. It’s compounded by the need to provide some contextual information … and by my prolixity 2. The latter is unavoidable, the former is probably necessary, not least because of the significant churn in new beekeepers.

A topic needs to be introduced, explained, justified and concluded.

Without this contextual information a post on oxalic acid trickling could be just:

5 ml of 3.2% w/v per seam when they’re broodless.

And where’s the fun in reading that?

Or writing it?

Furthermore, it’s probably of little use to a beginner who might not know what w/v means. Or what a seam is … or for that matter why being broodless is critical.

Keeping it topical

To maximise the income from site advertising I need to keep readers returning. This means the choice of topics should be important.

However, although some topics are chosen because they’re key concepts in the art and science of beekeeping, the majority are picked simply because I find them interesting.

And this week is one of the latter as I’m going to be droning on about … drones.

Specifically about drone numbers in the colony.

This was prompted by seeing the first drones of the season turfed out of the hive.

Dead drone at hive entrance

Another one bites the dust

Seeing this coincided with me discovering an interesting paper on how the queen’s laying history influences whether she produces drone or worker brood. This, inevitably, led me to other papers on drone production and discussions of how the colony controls drone numbers 3.

Drones are topical now because their days in your colonies are limited.

Already the colony will be producing significantly less drone brood than three months ago. The drones the colony has already produced will still 4 be flying strongly on good days.

However, when in the hive they will be being increasingly harassed by the workers.

Herding drones

If you open a colony very gently in the next few weeks 5 you might find the corners of the box contain high numbers of drones. The photo above was taken in late August and I’ve seen it several times late in the season. My interpretation is that it’s the only location in the hive in which drones can escape harassment by the workers.

Drone eviction

Drones ‘cost’ the colony a lot to maintain. A drone consumes about four times the amount of food than a worker (Winston, 1987). Therefore, once the fitness benefit of keeping drones falls below the expected costs needed to keep them they become ’surplus to requirements’. At this point the workers turf them out of the hive.

Evicted drones cannot feed themselves, so they perish.

It’s a tough life.

Interestingly, workers preferentially evict old drones. Presumably younger drones are more likely to fly strongly and mate with a virgin queen. Additionally, sperm viability in older drones is reduced, so their genes (and therefore those of the colony) are less likely to be passed on.

This ‘cost’ of maintaining drones is influenced by both the colony and the environment. For example, queenright colonies (which, by definition, have less need for drones) evict more drones than queenless colonies in the autumn, as nectar becomes limiting.

Although most beekeepers associate drone eviction with late summer/early autumn it also occurs when nectar is in short supply e.g. during the ‘June gap’.

It has also been suggested that drone eviction rates are related to colony size. Small queenright nucs, which have less need for drones, are more likely to evict than a full colony.

There’s still a lot we don’t know about drone eviction. For example, since drones tend to accumulate in queenless colonies, do these preferentially evict related drones to maintain potential genetic diversity in the population? 6

Hannibal the cannibal

Allowing an unfertilised egg to hatch, feeding the larva, incubating the pupa to emergence and then maintaining the resulting drone is a waste of resources if conditions are not appropriate. For example, doing this during a nectar dearth – particularly when drones are unlikely to be required for mating – makes no sense 7.

Therefore, in early spring and late autumn, workers cannibalise developing drone larvae. Effectively they are recycling colony resources. They preferentially cannibalise young larvae rather than older larvae. This makes sense as young larvae are going to need more food to reach maturity.

As above, queenless colonies cannibalise less queen-laid 8 drone larvae than queenright colonies.

In addition, some studies have shown that colonies with abundant adult drones cannibalise a greater proportion of developing drones. Again, this makes reasonable sense. Why rear more if you’ve got enough already?

However, to me it makes ‘reasonable’, but not ‘complete’ sense. Drones being reared as larvae are genetically related to the colony, adult drones may well not be. Drones that have drifted in from adjacent hives may therefore reduce the likelihood of the colony passing on its genes under environmental conditions which favour larval destruction but not eviction of adult drones.

Someone needs to look into this in a bit more detail 😉 .

There are lots of other aspects of larval cannibalism that are not understood. For example, how do workers discriminate between drone and worker larvae? Can they – as the queen can – measure the cell dimensions? Drone and worker brood pheromones differ from day 3 or 4. This seems a bit late to account for the cannibalisation of young larvae?

The influence of the queen

Since workers may cannibalise developing larvae 9 at different rates (drone vs. worker) it’s necessary to measure the colony’s egg sex allocation to see how the queen may influence drone numbers.

Only a few studies have done this …

There are experiments that suggest (they’re not definitive) that queens in continuously fed colonies lay more drone eggs in spring and summer than in autumn. This implies that day length or temperature may influence the queen, but it could also be a response to colony strength i.e. the queen lays more unfertilised eggs in a colony increasing in size, than in one decreasing.

In addition – and this is where I started down this rabbit hole in the first place – the egg laying history of the queen influences her current egg laying activity.

This easy-to-understand study was conducted by Katie Wharton and colleagues (Wharton, 2007). They confined queens for a period on either drone (DC) or worker comb (WC) – ensuring the queen could only lay drone or worker eggs for 4 days. They then transferred the queens to frames containing a 50:50 mix of drone and worker comb and recorded the amount of drone or worker eggs laid over 24 hours.

WC queens laid more drone eggs but the same amount of worker eggs as DC queens

There was a marked difference in the egg laying activity of the DC or WC queens when given the choice of laying drone or worker eggs. Although both the DC and WC queens laid similar amounts of worker eggs, the WC queens produced significantly more drone eggs as well.

Egg laying history or drone brood quantification?

This is a good study. The authors controlled for a variety of factors including season, colony size and food availability.

They additionally excluded the possibility that the egg laying activity of the queen was influenced by preferential cleaning of particular cells types by the workers, or by the workers backfilling certain cell types with nectar.

Finally, Wharton and colleagues allowed the colony to rear the eggs laid to pupation. The bias already observed was retained i.e. colonies headed by WC queens reared significantly more drone pupae than those headed by DC queens. The workers did not ‘correct’ the negative feedback exhibited by the WC queen, for example by preferentially cannibalising drone brood.

Although I termed this the ‘egg laying history’ of the queen a few paragraphs ago there is another interpretation.

The worker or drone comb already laid up by the queen – during the 4 day confinement period – remained in the colony. It’s therefore possible that the egg laying activity of the queen was influenced by the amount of drone brood already present in the colony.

Either explanation is intriguing.

How does the queen ‘count’ the number of drone or worker eggs she’s laid in the recent past? Alternatively, how does she quantify the amount of drone brood in the colony?

But what about the workers?

The Wharton study largely excluded the possibility that there was preferential cleaning of drone or worker cells by the workers in the hive. In fact, earlier studies have indicated that cell cleaning continues almost constantly and workers were equally likely to clean worker or drone cells.

The Wharton study also addressed – and excluded – the possibility that differences in the backfilling of drone or worker cells might influence egg laying.

However, it’s not understood what determines whether drone cells get backfilled with nectar by workers. My colonies are starting to do this now. Do the workers fill drone cells with nectar because the queen hasn’t laid in these cells or because they only backfill drone cells late in the season?

The former suggests that there is some sort of competition between the egg laying activity by the queen and nectar storage by workers. In contrast, the latter suggests that there are environmental triggers that influence this worker activity.

Or both … of course 😉 .

Comb building

In contrast to some of the studies outlined above, comb building is easy to monitor and – perhaps consequently 10 – has been well studied.

I’ve already discussed comb building in a recent post about queenless colonies. These preferentially build drone comb (Smith, 2018).

What else influences drone comb production?

Probably the two strongest determinants are the amount of drone comb already in the nest and the season.

Drone comb production is reduced in colonies that already contain lots of drone comb. Many beekeepers never observe this as they only use frames containing worker foundation. The workers squeeze in little patches of drone comb – often in the corners of the frame – but it never exceeds 5% of the total.

Colonies often prefer to build drone comb when given the choice

In contrast, natural nests contain 15-20% drone comb. That’s equivalent to two full frames in a National-sized hive. Once drone comb approaches this level a negative feedback loop operates and the workers build less drone comb. The negative influence of this drone comb (on building more drone comb) is enhanced if the comb contains drone brood.

Colonies drawing comb now (and certainly in the next month or two) will build worker comb. Some beekeepers exploit this to get lovely new worker frames drawn – nucs are particularly good at this 11. In contrast, drone comb is drawn in spring and early summer. The season – presumably day length and temperature – therefore influences drone comb production, and hence drone production.

A thousand words

Well, nearer 2000.

As we near the end of the season we start to see drones evicted from our colonies. It’s interesting to think about the interplay of events that resulted in the colony producing those drones in the first place … and how and why the colony regulates drone production throughout the season.

Wharton (2007) neatly summarised the five stages from comb building to adult drone eviction.

Drone production and maintenance in a honeybee colony

I’ve dealt with these in reverse order because that was the best fit with the photo of the dead drone on the landing board that I started with.

There’s a lot we still don’t understand about the regulation of drone numbers. In particular, I think the majority of studies have ignored the influence of adult drone numbers on any of five stages illustrated above.

This is an important omission as drones move more or less freely between hives. That means that adult drones may well be genetically unrelated to the colony.

Perhaps this means that adult drones do not influence drone production? After all, if they did negatively influence drone production – as suggested above – it would potentially limit the ability of a colony to reproduce its genes. Evolutionarily this doesn’t make sense (at least, to me).

There are a couple of studies that have tried to determine the influence of adult drones, but they have produced conflicting results. Rinderer (1985) added drones to a colony which consequently reduced drone brood production. However, Henderson (1994) did the opposite and showed that removal of adult drones had no effect on drone brood production.

There’s clearly lots more to do …


Notes

I wrote this late on Thursday night. While doing so I watched the page views of my four year old post on Mad honey go ‘off the scale’ (which for a beekeeping site means hundreds of views per hour). The interest wasn’t sparked by my erudite description of grayanotoxin intoxication. Instead it was related to a video of a ‘stoned’ Turkish brown bear cub rescued after eating honey produced from rhododendron nectar.

It’s now abundantly clear that if I want to maintain my outrageous advertising income I should probably write more about hallucinogenic honey and less about the evolutionary subtleties of honey bee sex ratio determination.

That’ll teach me 😉

References

Boes, K.E. (2010) ‘Honeybee colony drone production and maintenance in accordance with environmental factors: an interplay of queen and worker decisions’, Insectes Sociaux, 57(1), pp. 1–9. Available at: https://doi.org/10.1007/s00040-009-0046-9.
Henderson, C.E. (1994) ‘Influence of the presence of adult drones on the further production of drones in honey bee (Apis mellifera L) colonies’, Apidologie, 25(1), pp. 31–37. Available at: https://doi.org/10.1051/apido:19940104.
Rinderer, T.E. et al. (1985) ‘Male Reproductive Parasitism: A Factor in the Africanization of European Honey-Bee Populations’, Science, 228(4703), pp. 1119–1121. Available at: https://doi.org/10.1126/science.228.4703.1119.
Smith, M.L. (2018) ‘Queenless honey bees build infrastructure for direct reproduction until their new queen proves her worth’, Evolution, 72(12), pp. 2810–2817. Available at: https://doi.org/10.1111/evo.13628.
Wharton, K.E. et al. (2007) ‘The honeybee queen influences the regulation of colony drone production’, Behavioral Ecology, 18(6), pp. 1092–1099. Available at: https://doi.org/10.1093/beheco/arm086.
Winston, M.L. (1987) The Biology of the Honey Bee. Cambridge, Massachusetts: Harvard University Press.

Shook swarms and miticides

Synopsis : Combining a shook swarm with miticide treatment removes most mites in the colony and dramatically reduces DWV levels. The application of this strategy for practical beekeeping is discussed.

Introduction

Why does Varroa have such a devastating impact on colony health?

Feeding on haemolymph – or the abdominal fat body – by Varroa is probably detrimental. Furthermore, during feeding the mite induces immunosuppressive responses which make the bee both more susceptible to bacterial infections and compromises its nutritional status (Aronstein et al., 2012 1 ).

But if that wasn’t enough, the real damage is caused by transmission of viruses – in particular deformed wing virus (DWV) – from the mite to the developing pupa (and adult worker, as mites probably also feed on newly eclosed workers during the misnamed phoretic stage of the life cycle).

In the absence of Varroa, DWV is seemingly inconsequential for honey bees. Varroa-free colonies – including mine on the remote west coast of Scotland – carry DWV, but virus levels are very low and there is never any overt disease.

But Varroa infested colonies, particularly at this time of the season, often have very high levels of DWV.

Individual pupae parasitised by Varroa can develop stratospherically high DWV levels – reaching over a million times higher levels than seen in unparasitised bees (which can be similar to those recorded in Varroa-free bees). In the mite-exposed pupae the virus levels can kill the developing bees, or result in the characteristic symptoms (primarily deformed wings but also stunted abdomens and discolouration) that give the virus its name.

Worker bee with DWV symptoms

Worker bee with DWV symptoms

But bees not directly exposed to Varroa also have higher DWV levels in mite-infested colonies, particularly as the season progresses. Presumably this is due to horizontal transmission of the virus during larval feeding or trophallaxis.

What happens to these elevated virus levels after the removal of Varroa using a miticide such as Apivar?

Who cares? … I mean, Why could that matter?

The clue is in the section above.

Here it is again:

But bees not directly exposed to Varroa also have higher DWV levels [ … snip … ] presumably this is due to horizontal transmission of the virus during larval feeding or trophallaxis.

If you remove mites the virus levels in the treated adult bees are often surprisingly high 2. That makes sense because the miticide is only removing the vector for the virus … the bees with high levels of virus infection are unaffected.

If, during larval feeding or trophallaxis, these elevated levels of DWV result in yet more bees acquiring high DWV levels then the health of the colony will remain compromised.

The real reason that DWV is a problem for honey bees is that high levels of the virus result in the reduced longevity of bees. This isn’t an issue for the short-lived summer foragers 3. However, reducing the longevity of the winter bees – the so-called diutinus bees – can be fatal for the colony. These are the bees that support the queen in winter, thermoregulating the hive and that rear the first brood of the following season.

Their importance to successful overwintering cannot be overemphasised.

So, the question remains. What happens to the virus levels in the hive after the removal of Varroa?

Of course, the reason I’m posing this question is that we now know … 😉 .

Two easy-to-understand potential outcomes

It seemed to us that there were at least two likely outcomes.

  1. The virus levels in the hive drop very quickly after mite removal (red dashed line, below) and return to some sort of basal level. How quickly and to what basal level? We didn’t know.
  2. Virus levels remain elevated for a long period after Varroa is removed (red solid line, below). How long and to what elevated level? Yes – you guessed it – we didn’t know 😉 .

Of course, biology isn’t binary. There are any number of alternative outcomes … it’s just that those two seemed the most likely.

Two possible outcomes for virus levels after mite removal (black vertical dashed line)

What’s more, they’re the easiest to understand … and to explain.

Why might virus levels remain high if Varroa are removed?

Surely the short lifespan of adult bees means these would soon be lost from the colony … particularly if they have reduced longevity?

Yes, but …

We published a paper a couple of years ago that clearly demonstrated that honey bee larvae fed high levels of DWV became infected with the fed virus. The latter, which we could distinguish from any DWV already present in the larvae, replicated to similar high levels seen in a mite-infested hive (Gusachenko et al., 2020).

This observation perhaps suggested that the second scenario outlined above could occur. All the mites are slaughtered, but the remaining bees with high levels of DWV feed developing brood which consequently also go on to develop high levels of DWV.

Although it’s always good to remove mites this would not be the best outcome for the colony.

Virus quantification

Before I explain how we tested which, if any, of these two possibilities is correct I need to say a few things about virus ‘levels’.

For a variety of reasons I don’t have time, space or energy to explain, we don’t actually count viruses, instead we count copies of the virus’s genetic material (the genome).

All the magic happens in one of these machines – a Bio-Rad CFX96 Touch Real Time PCR system.

The virus genome is made of ribonucleic acid (RNA) and we can therefore use fantastically expensive sensitive and accurate diagnostic methods to measure how many copies are present in a particular sample – for example, in a worker bee, or a developing pupa.

Still with me?

Good.

To complicate things a little, we can’t meaningfully express the number of virus genomes present as an absolute number (like one million, or 2,478) because bees are different sizes; larvae are tiny, pupae are bigger, drones are larger still.

In addition, different workers are different sizes, larvae grow etc.

Therefore we express it as genomes per unit of total RNA extracted from the sample. That’s a bit of a mouthful, so we abbreviate it to GE / μg 4.

Phew!

And finally, to put some numbers on the low and high levels of DWV I discussed earlier, a bee from a Varroa-free colony contains ~1,000 – 10,000 GE / μg (103 – 104) of DWV whereas a pupa parasitised by Varroa regularly has 10,000,000,000 to 1,000,000,000,000 GE / μg (1010 – 1012).

That’s a lot of virus 🙁 .

The experiments

Experiments plural because we did these studies in both 2018 and 2019. ‘We’ are Luke (a then PhD student and now post-doctoral fellow in my laboratory, and the first author on the paper) together with our friends and collaborators, Craig, Ewan and Alan (in Aberdeen) and Giles (in Newcastle). The work was published a few days ago in the journal Viruses and is ‘open access’ (Woodford et al., 2022). This means that anyone feeling particularly masochistic or suffering from sleep deprivation can read all the gruesome details at their leisure.

Not ‘breaking rocks in the hot sun’ … but it sometimes feels like that

The paper covers more than just the one experiment I’m going to discuss here. We also looked at how the virus population changes when mite-free bees become infested with Varroa.

I’ll save that for another post 5  … it’s a good story in its own right.

Most mites are in capped cells

It’s been known for at least three decades that the majority of the Varroa population in a brood rearing colony are within capped cells, feasting on developing pupae.

Nom, nom, nom!

Precisely what percentage of the population is the majority varies a bit 6, but a figure of 90% is often quoted as typical for midseason.

% of mites in capped cells

The percentage of mites in capped cells (this is predicted, not actual data)

We reasoned that the best way to quickly remove all 7 the Varroa in a colony was to combine treatment of the phoretic mites with removal of all the brood … where the majority of the mites are lurking.

And to remove the brood (and associated mites) we conducted a shook swarm.

The shook swarm

Many beekeepers will be familiar with the technique called a shook swarm.

Shook swarm setup. Note Apivar strips in the open hive. Returning foragers already clustering at the entrance

This involves shaking all the adult bees into a new hive with frames containing fresh foundation. All the old frames and brood from the original hive are discarded.

We modified this by including Apivar strips in the hive into which we shook the adult bees.

Shook swarmed colony strapped up for transport … we wait for all the bees to enter the hive before moving it

The ‘shook swarm and miticide’ experiment – which we conducted in May – therefore involved the following steps (we used three strong double brood hives per season, each containing similar amounts of bees and brood):

  1. We quantified DWV in emerging brood in hives in which no Varroa management was conducted.
  2. The queen was removed, caged and kept safe for a few hours.
  3. All adult bees were shaken into a new brood box containing 11 frames of fresh foundation and two strips of Apivar 8.
  4. The shook swarms were relocated to a quarantine apiary.
  5. The queen was returned to the shook swarmed colonies and they were fed ad libitum with syrup to encourage them to draw fresh comb.
  6. Mite drop was recorded at 5 day intervals, increasing to longer intervals, until October when brood rearing ceased.
  7. DWV levels were quantified on a monthly basis from June to October.

As you can see, a very simple experiment.

The results

The mite levels in the ‘donor’ hives were much higher in 2019 than 2018. It’s not unusual to see this type of year to year variation in mite levels. In this instance the mean temperature in February and March 2018 had been several degrees colder than 2019 (remember the Beast from the East?).

The Beast from the East ...

The Beast from the East …

This almost certainly reduced early season brood rearing and so delayed mite replication. Brood rearing was strong by late Spring, but the mite levels in 2018 had yet to catch up.

The results of the experiment in both years were essentially the same. However, for clarity I’ll just present the 2019 data as the mite infestation numbers were so dramatic.

Mite drop after conducting the shook swarm

The cumulative mite drop from Apivar-treated shook swarms ranged from ~500 to ~3000 in the first 5 days. After that the daily mite drop remained at extremely low levels until recording stopped in October.

Mite drop following shook swarm and Apivar treatment

If you assume that only 10% of mites were phoretic at the time we conducted the shook swarm, this means that the total number of mites in some of these colonies was about 30,000. Even the colony with the lowest mite drop may have been hiding an additional 4,500 mites in capped cells.

Remember … the National Bee Unit guidance states that if mite levels exceed 1,000 then treatment is strongly recommended ’to avoid Varroa causing significant adverse effects to the colony’.

I think this part of the study shows just how effective Apivar is. After the first 5 days of treatment the cumulative drop – the Apivar strips still were left in place for 8 weeks – was extremely low for each fortnightly sampling period.

Of course – other than the very high numbers – none of this was particularly surprising. We know Apivar kills Varroa.

Perhaps you’re thinking ”My hives drop more Varroa during the autumn treatment, and for longer.”

When you treat a colony with brood present the mite drop is high in the first few days, but then often remains significant over the next 2-3 weeks while the mite-infested brood emerges. 

In our case, all the mites were on adult bees. By killing these mites in the first few days before there was new sealed brood in the colony we ensured the majority of the new brood did not become infested.

Virus levels before and after the shook swarm

In each colony we sampled a dozen emerging workers, once before the shook swarm and then on a monthly basis until brood rearing stopped. By testing emerging brood we could be certain they had been reared in the test colony, rather than drifting in from elsewhere. 

Before the shook swarm virus levels ranged from 105 to 1010 per worker, with an average of around 5 x 107 GE / μg. For those of you unfamiliar with scientific notation that is 50 million virus genomes.

Virus quantification in individual workers from colonies before and after the shook swarm and Apivar treatment

Strikingly, from the June sample onwards, virus levels dropped to an average of about 104 GE / μg (10,000 virus genomes, a 5,000-fold reduction). This average obscured a range of individual levels, from about 102 to 106.

These reductions are statistically significant … always reassuring 😉 .

The 2018 data showed a similar marked reduction in virus levels. The pre-treatment levels were marginally lower (remember, it was a ’low Varroa’ season), but the levels dropped to an average of only 1,000 GE / μg, a slightly higher fold-reduction and again highly statistically significant.

If you remove the majority of the Varroa the virus levels drop very fast to levels seen in mite-free colonies, or colonies with very low mite counts.

Tough love?

Some beekeepers consider that a shook swarm is tough on the colony. 

I’m not sure I agree.

How and when the shook swarm is done matters a lot.

It can be tough, but it shouldn’t be.

The bees need to draw new comb. For this they need ample feeding, lots of bees and warm weather. By conducting shook swarms on strong colonies in late May and giving them a few gallons of syrup we achieved all this.

‘I know I put that caged queen down here … somewhere’

Doing a shook swarm on a weak colony, too early (or late) in the season or omitting feeding is a recipe for disaster. The colony will struggle to draw comb, its brood rearing will be limited and it will be playing ’catch up’ for the remainder of the year.

Our shook swarmed colonies were booming by late July and entered the winter very strong. All overwintered successfully.

I’d argue that a shook swarm is a lot less tough on a colony than the disease burden caused by thousands of mites … 🙁 .

Why Apivar?

It’s worth emphasising that this was a scientific experiment to investigate the consequences for the virus population of removing almost all of the Varroa.

It was not designed as an example of how a beekeeper would necessarily choose to manage a honey production colony.

Our choice of Apivar was considered and deliberate. Application is straightforward, toxicity – at the levels we used – is undetectable and, critically for these studies, it remains active for weeks.

Apivar strip on wire hangar

Of course, Apivar cannot be used when there are honey supers on the hive 9. Any supers added for the summer nectar flow were not extracted.

Additionally, feeding gallons of syrup when there are honey supers present is also not recommended 😉 .

What else could we have used?

The two obvious choices were MAQS or oxalic acid. Both are effective against phoretic mites, though perhaps less so than Apivar. However, both are only active for a short period in the hive; the treatment period for MAQS is 7 days and the activity of oxalic acid – trickled or vaporised – is probably less than a week.

Neither could be relied upon to slaughter the maximum number of mites, a necessity to produce an understandable result 10. We were additionally concerned about problems with queens or absconding had we used MAQS (both of which would have invalidated the study), and we were keen to avoid the need for repeat treatments with oxalic acid (not least because this is not an approved application method).

With thousands of mites we wanted to ensure that the majority were killed quickly … and, as important, that any that survived the first few days of miticide treatment were also more than likely to be killed later 11.

Application to practical beekeeping

The main aim of this experiment was to investigate the levels of DWV in the colony after the majority of Varroa are removed. However, we were also mindful that the method may be useful for a beekeeper who discovers his/her colony has damagingly high mite levels mid-season, or for someone who inherits abandoned hives with high mite loads.

In these scenarios, assuming there are sufficient bees, some nice warm weather and lashings of syrup available, the combination of a shook swarm and simultaneous miticide application is probably the fastest way to restore colony health.

I am not suggesting that beekeepers routinely conduct a shook swarm and miticide application mid-season. It might not be tough on the colony, but that doesn’t mean it’s not very disruptive. If it’s not needed (because mite levels are well controlled, for example) then it’s a waste of brood … and syrup.

However, there are times when I could imagine it might be useful.

If your primary crop is heather honey you’ll know that the hives sometimes don’t come back from the hills until late-September. That’s late to be applying miticides to protect the winter bees. In an area with an extended June gap (which often starts in May) it might be possible to effectively rid the hives of Varroa in June and have a strong colony to take to the moors in early August.

This is probably a better approach than using a half dose of Apivar in June (as some do) which probably doesn’t kill all the mites anyway, risks contributing to amitraz resistance in the mite population and may result in Apivar strips being left in the hive during the heather flow 12.

Conclusions

Miticides kill mites … big deal.

However, it’s the viruses – in particular deformed wing virus – that kill colonies.

We have now shown that removing the majority of the mites from a colony (including those associated with sealed brood) results in the levels of DWV in the hive dropping very quickly.

The speed with which this happens – four weeks or less – is probably accounted for by the lifespan of the adult bees in the colony following the shook swarm.

This suggests that high levels of virus are not horizontally transmitted or (and this is subtly different) that horizontal transmission, through feeding, of large amounts of virus does not result in elevated levels of virus replication in the recipient bee (larva or adult).

All sorts of questions remain. Would oxalic acid be a suitable replacement for Apivar? How much virus is transferred from a worker to a larva during brood rearing, or between workers during trophallaxis? Is this below a threshold for efficient infection? Do virus levels drop as dramatically when treating a broodless colony (e.g. after caging the queen for three weeks)?

In the meantime just remember that ”the only good mite is a dead mite” … and, if you kill the mites, you also quickly reduce virus levels to a level at which they do not damage the colony.

And a straightforward way to achieve that is to combine a shook swarm with an effective miticide.

Result!


References

Aronstein, Katherine A., Eduardo Saldivar, Rodrigo Vega, Stephanie Westmiller, and Angela E. Douglas. ‘How Varroa Parasitism Affects the Immunological and Nutritional Status of the Honey Bee, Apis Mellifera’. Insects 3, no. 3 (27 June 2012): 601–15. https://doi.org/10.3390/insects3030601.

Gusachenko, Olesya N., Luke Woodford, Katharin Balbirnie-Cumming, Ewan M. Campbell, Craig R. Christie, Alan S. Bowman, and David J. Evans. ‘Green Bees: Reverse Genetic Analysis of Deformed Wing Virus Transmission, Replication, and Tropism’. Viruses 12, no. 5 (May 2020): 532. https://doi.org/10.3390/v12050532.

Woodford, Luke, Craig R. Christie, Ewan M. Campbell, Giles E. Budge, Alan S. Bowman, and David J. Evans. ‘Quantitative and Qualitative Changes in the Deformed Wing Virus Population in Honey Bees Associated with the Introduction or Removal of Varroa Destructor’. Viruses 14, no. 8 (August 2022): 1597. https://doi.org/10.3390/v14081597.

 

 

Workers not shirkers

Synopsis : Not all foragers are equal. A small proportion – the elite foragers – make the majority of foraging trips. These are the most experienced foragers. Could pathogens and pesticides that reduce worker longevity compromise nutrition of the hive?

Introduction

All bees are the same.

Right?

No, of course not.

The three castes, from The ABC of Bee Culture, 1895

For a start there are three castes of honey bee; the queen, drones and workers … but we can also sub-divide these castes.

Queens

For example, most beekeepers would agree that there are fundamental and important differences between a virgin queen and a mated queen. They behave differently, their physiology is different and so are their their senses.

Drones

Similarly, the difference between virgin and mated drones is also pretty fundamental. In fact, it’s literally a matter of life and death 😉 . However, there are also less dramatic – largely physiological – differences between sexually immature and mature drones.

Workers

And there are differences in this caste as well.

Any beekeeper who uses Pagden’s artificial swarm for their swarm control has – although perhaps unknowingly – exploited the difference between two broad groups of workers; the hive (or nurse) bees and the flying bees (or foragers). The former are bees that have yet to fly from the hive. They rear the developing brood, look after the queen and perform a range of housekeeping duties.

After about three weeks the maturing worker goes on several orientation flights and eventually becomes a forager, responsible for collecting the pollen, nectar, resin and water the colony needs.

‘Eventually’ because workers undertake additional roles e.g. guard bees, undertaker bees and scouts, as they segue from hive bees to foragers 1. This change in roles during the lifetime of a worker bee is termed temporal polyethism 2.

Elite foragers

In this post I’m going to focus on the last of the roles the worker fulfils, that of foraging.

There is a lot of good observational and experimental science on foraging behaviour; for example, the preference of foragers for certain pollen or nectar sources, or the features of the colony that induces foraging activity. Some of this is briefly reviewed in ’A closer look – Foraging behaviour’ by Clarence Collison in Bee Culture 3.

Instead of rehashing those things I’m instead going to describe the concept of ’elite’ foragers. These are a minority of the forager population that do the majority of the foraging. They are therefore probably a very important cohort of bees for the colony.

The definition of elite foragers was first demonstrated for honey bees in studies conducted by Paul Tenczar working with Gene Robinson in 2014 4.

Gross differences in foraging activity was not a new concept. It had been observed in a wide range of eusocial insects in studies dating back to the 1970’s. Since reproductive fitness of eusocial insects – like bees, wasps and ants – is determined at the colony level, and workers are genetically related, variation in worker performance was neither expected nor had an obvious origin.

However, significant differences in worker performance are observed when suitable technology exists to detect it.

’We have the technology’ 5

Tenczar used RFID tags to label individual worker bees. I’ve described this technology before. It allows the unique identification of individual bees. Foragers were detected leaving or arriving at the hive by monitoring them with two RFID ‘readers’ arranged along the narrow hive entrance tunnel.

Theoretically at least, a bee registered by the inner and then the outer reader should be leaving the hive, whereas one registered first by the outer reader, followed by the inner, would be arriving. If these two pairs of events were separated by a several minutes it should mean the bee has successfully completed a round trip.

Unfortunately, the RFID/reader technology was in its (relative) infancy 6 and trips were missed. They even added two RFID chips to each bee to improve detection rates. Manual observation showed that there was a 76-94% chance of a trip being detected by at least one of the four readers. They therefore used reads rather than trips as a metric for activity level. This is a bit of a fudge but it will do for the purpose of this study.

As I will show shortly, the technology has now improved and there are more accurate ways to measure foraging trips.

Orientation flights and the age of onset of foraging

Tenczar et al., labelled over 1000 day-old workers in five separate experimental colonies and monitored their activity for 5-7 weeks.

Orientation flights occur before foraging flights, and usually take place in the afternoon. To be sure they were only monitoring foraging flights, they defined the first day of foraging activity as the one when there were at least 6 ‘reads’, and with at least 25% of reads occurring before midday.

Age at onset of foraging for tagged bees

The average age of a worker at the onset of foraging was 20.4 days – a figure in agreement with the ‘three weeks in the hive’ statement I made above. However, if you average the five separate lines of hive data (above) it is also clear that a significant proportion of the bees (actually 27% of them) started foraging within the first 10 days.

This is so-called ’precocious foraging’ and had been seen previously in colonies created with a single cohort of bees. The colonies used in these studies were small (~1000 bees in each) and were started with bees of all the same age. The fact that some start foraging precociously is a demonstration of the plasticity in temporal polyethism I referred to in an earlier footnote … and which non-scientists would describe as doing ‘different jobs at different times, that might vary’.

Workers and shirkers

However, just looking at the cumulative count of workers (above) it is clear that there is considerable variation in the timing of the onset of foraging.

In addition, and perhaps more surprisingly, the level of foraging activity also varied greatly.

Some workers made rather few foraging trips, others started early and finished late, making repeated closely-spaced trips throughout the day.

Contributions of individual bees to the total foraging activity in two colonies

These histograms show the relative foraging activity of RFID tagged bees in two representative colonies. The vertical axis records the number of bees making a particular relative foraging effort. The shape of the graph – large bars on the left and much smaller bars on the right – show that the majority of the bees (hence the large bars) do relatively little foraging, whereas a much smaller number of bees (in the shorter bars on the right) do lots.

Lorenz curves

A more informative way to represent this data is to use a Lorenz curve which displays the share of foraging activity (vertical axis) against the percentage of foraging bees (horizontal axis).

Example plot of a typical Lorenz curve of cumulative share of foraging activity for one of five study colonies

If all foragers contributed equally to foraging activity the ‘curve’ would be the diagonal dotted line i.e. 50% of the bees would ‘deliver’ 50% of the foraging activity.

In the colonies studied, the actual contribution to foraging activity is shown by the curved line.

Approximately 20% of the foragers accounted for 50% of the foraging activity (the area I’ve shaded blue). These are the elite foragers. Conversely, the ‘laziest’ 20% of foragers make less than 5% of the foraging trips (shaded red) 7.

These are the shirkers … the less said about them the better 😉 .

Tenczar et al., conducted additional analysis of the pattern of foraging activity per bee per day, and the consequences of removal of elite foragers (in due course other foragers become elite foragers). However, I’ll skip these as I want to move on to a more recent study of elite foragers.

We really do ‘have the technology’

In 2019 Simon Klein and colleagues published another RFID-tagged forager study 8. In the intervening years the RFID tag and reader technology had improved. They used two modified four frame nucleus hives in which bees traversed separate tunnels for entry and exit.

Colony entrance with sensors – bees enter and depart the hive using different tunnels

In addition to using more reliable RFID readers (#3 in the diagram above) they also weighed (#4) the bees as they entered and exited the hive and recorded video (#7) of the returning bees to determine whether they were carrying pollen.

These additional measurements meant that, in addition to the number of trips completed, the authors were also able to measure foraging performance in terms of pollen or nectar collected.

Actually, that’s a bit of an overstatement.

Pollen foragers were identifiable on video by their pollen-filled corbiculae (PDF). In contrast, foragers returning without pollen could have made unsuccessful trips, or may have collected nectar or water. They therefore classified foraging activity into ‘pollen’ or ‘non-pollen’ trips.

In total they monitored 564 foragers who made an average of 19 foraging trips in their lifetime (~10,500 trips in total). Interestingly, the average foraging lifetime was less than 5 days. As with Tenczar et al., they excluded orientation flights from the trips recorded (though in a different way).

Practice makes perfect

None of the bees monitored foraged exclusively for pollen. However, it was noted that the more trips a bee made, the greater the proportion of the trips were for pollen until a maximum was reached, after which pollen collecting declined.

Changes of foraging performance with experience – pollen collection (each line is an individual tagged bee)

Foragers lost weight on pollen foraging trips – presumably using crop contents to ‘fuel’ the flight. Since the weight of the crop contents were unknown, it wasn’t possible to calculate the weight of the pollen collected.

Non-pollen foragers weighed about the same (or a little more) upon return as when they left. Since both the weight of the crop contents and the identity of what was being collected (water or nectar) were unknown, it was not possible to determine how much had been collected. However, as individual bees aged – and so took more non-pollen foraging trips – their gain in weight per trip increased. This suggested that (as with the likelihood of collecting pollen) increased experience resulted in more efficient foraging.

The elite foragers are the best performing foragers

Analysis of the average number of foraging trips per day demonstrated that it increased over the first ten days and then plateaued at about 10 trips per day.

Changes of foraging performance with experience – average trips per day

It’s worth noting a couple of points here; firstly, on average foragers only made 19 trips in their lifetime and secondly, the majority of the foragers never reached maximal pollen foraging activity (in the multicoloured graph above, most lines terminate before they reach a peak and start to decline).

These points, together with the average foraging lifetime being under five days, indicate that there is a very high attrition rate amongst foragers.

Most die young … by which I mean within the first week of leaving the hive 🙁 .

But, some live long enough to become the experienced elite foragers, and these bees were the best performing foragers.

Like the previous study, an average of 19% of the foragers performed 50% of all foraging trips recorded 9. These were the elite bees.

These elite bees were the most likely to collect pollen and – on non-pollen trips – were most likely to show a gain in weight, indicating a greater resource (nectar or water) load was being carried.

Conclusions and consequences

The logical conclusion is that, through experience, bees improve their foraging performance. However, most bees never realise their full potential as they perish long before they achieve the status of elite foragers.

It is known that bees exhibit both learning and memory. With regard to foraging, it’s known that navigation skills improve with experience, and that both flower discrimination and ‘handling’ also get better i.e. they are more likely to find (and re-find) remote flowers, to distinguish them from other flowers in the immediate area and to harvest the pollen or nectar from the flower.

These elite bees must make a significant contribution to resourcing the colony.

Numerically they are a minority of the foraging population, but they collect lots of the pollen, nectar, water and propolis needed by the colony.

However, without knowing the precise number and age/experience distribution of the foragers and the mass of the pollen/nectar loads collected, it is not possible to determine whether they collect the majority of these resources.

For example, do 1000 inexperienced foragers collect more (or less) than 100 elite foragers? Are the massed ranks of young naïve foragers more effective at provisioning the colony than a few dozen of ‘old timers’?

We don’t know … yet.

The experiments needed to determine this are more difficult, and a lot more intrusive. You need to know both the weight of whatever was collected, together – in the case of pollen and nectar – with its value to the colony. For example, we know that bees preferentially forage on particular high protein pollens, or on high-sucrose nectar sources … presumably (though it needs to be shown) elite bees do this better than naïve foragers.

Looking after the elderly

Tenczar et al., showed that depleting the elite bee population had little long-term effect, because younger bees made additional foraging trips in subsequent days. However, this ‘replacement’ was only measured in terms of foraging trips, not foraging efficiency (which Tenczar didn’t measure).

If efficiency comes with experience – as suggested – it may be that additional time would also be needed to turn these extra flights into foraging trips that significantly benefitted the colony.

All of which means that stressors that adversely affect ageing bees, or that shorten the lifespan of foragers, may have a marked impact on colony pollen and nectar collection.

And there are lots of these sorts of stressors … pesticides, pollution, poor nutrition and pathogens – an alliterative gamut of threats to these important, elderly but highly effective bees. For example, increasing cumulative exposure to sub-lethal levels of pesticides may be deleterious to older bees.

Deformed wing virus

Unsurprisingly, being a virologist, it is the pathogens that interest me. In particular, deformed wing virus (DWV).

DWV symptoms

DWV symptoms

DWV is probably responsible for the majority of overwintering colony losses because it reduces the longevity of the (nominally long-lived) winter bees. I’ve discussed this at length elsewhere but the important bits are as follows:

  • winter bees should live for months, not weeks, maintaining the colony through until springtime
  • if there are lots of Varroa present during the early autumn (when winter bees are being reared) the developing winter bees will have high levels of DWV
  • some bees will die before emergence, but those that don’t will instead die in weeks, not months
  • consequently the winter cluster shrinks rapidly in size, becomes unable to thermoregulate and is separated from its stores
  • this doesn’t end well … the colony either dies, or struggles through to the spring and is too weak to expand

But what happens to foragers with high levels of DWV in the summer?

Studies from my lab 10 have shown that pupae injected with DWV – essentially recapitulating what Varroa does when it feeds on a developing pupa – have three potential fates; they either die during development (~15%; see B in the figure below), emerge with developmental abnormalities (~65%; the deformed wing bit which ’does what it says on the tin’) or emerge and appear ‘normal’ (~20%).

Unanswered questions

Interestingly, the small proportion of bees that appear ‘normal’ have indistinguishable levels of DWV to those that have deformed wings (panel A below).

The fate of bees injected with DWV

Do these bees with high-DWV levels live long enough to become foragers?

We don’t know.

If they do become foragers – which , frankly, I doubt – do they learn how to forage well?

Again, we don’t know, though we do know from other studies that high levels of DWV leads to some cognitive impairment, so navigation at least may well be suspect 11.

Finally, if they do become foragers, do they live a long and healthy life, or do they die prematurely?

Unfortunately … we don’t know this either.

Hive inspections

That’s a lot of ‘ifs’ there … and just as many unanswered questions.

Let’s assume that bees with high levels of DWV can mature to become foragers, but that they exhibit reduced longevity. If that is the case then the elite forager population would be reduced, so jeopardising provisioning the colony with nectar and pollen.

I’m sceptical that bees with such high DWV levels can survive long enough to mature into elite foragers. Nevertheless, I’d prefer to test this experimentally in the comfort of my lab 12, rather than in my honey-production or queen-rearing colonies.

Therefore, during hive inspections I look carefully for the signs of overt DWV disease or varroosis – bees with deformed wings, uncapped developing brood or phoretic mites. I also periodically measure mite drop. If I see see problems (and with correct timing and appropriate treatment in autumn and winter you shouldn’t 13 ) I intervene.

Midseason mite management may save the elite foragers … and help prevent the loss of the colony overwinter.

A gradation of DWV levels

However, there’s a related – more subtle – thing to consider.

Studies from our group (and others) have shown that injection of tiny amounts of DWV results in a very rapid replication of DWV to stratospherically high levels. As shown above, these kill or maim ~80% of exposed bees.

But there’s a less well understood feature of colonies with high Varroa levels. During the course of the season the levels of DWV in bees not exposed to Varroa during development rise.

In March or April, DWV levels may be ~103/bee 14. This is about the lowest level we ever see in bees, and is equivalent to the levels of DWV present in colonies from Varroa-free regions like Colonsay.

However, by mid- or late-summer the levels are 100-1000 times higher i.e. 105-106/bee 15. This is still 10,000 times lower than the levels DWV reaches in Varroa-parasitised pupae 16.

As an aside, we don’t formally understand how the DWV levels increase during the season. I suspect it’s through trophallaxis though we have also published some evidence of larval susceptibility to DWV during feeding.

Whatever the mechanism, bees carrying one million copies of DWV look completely normal and, as far as we can tell, behave completely normally.

‘As far as we can tell’, as no one has really done the right experiments …

I think it would be very interesting to carefully investigate the longevity and ability to achieve elite forager status for early season (very low DWV levels) and midsummer (intermediate DWV levels) bees.

Perhaps these intermediate levels of DWV are damaging after all?


Note

The Klein et al., paper has been very poorly proofread 17 and contains several errors, some of which potentially change the meaning of the text.