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OPEN The prediction of swarming in

honeybee colonies using vibrational
spectra
Michael-Thomas Ramsey1, Martin Bencsik1 ✉, Michael Ian Newton1, Maritza Reyes2,
Maryline Pioz2, Didier Crauser2, Noa Simon Delso3 & Yves Le Conte2
In this work, we disclose a non-invasive method for the monitoring and predicting of the swarming
process within honeybee colonies, using vibro-acoustic information. Two machine learning algorithms
are presented for the prediction of swarming, based on vibration data recorded using accelerometers
placed in the heart of honeybee hives. Both algorithms successfully discriminate between colonies
intending and not intending to swarm with a high degree of accuracy, over 90% for each method, with
successful swarming prediction up to 30 days prior to the event. We show that instantaneous vibrational
spectra predict the swarming within the swarming season only, and that this limitation can be lifted
provided that the history of the evolution of the spectra is accounted for. We also disclose queen toots
and quacks, showing statistics of the occurrence of queen pipes over the entire swarming season. From
this we were able to determine that (1) tooting always precedes quacking, (2) under natural conditions
there is a 4 to 7 day period without queen tooting following the exit of the primary swarm, and (3)
human intervention, such as queen clipping and the opening of a hive, causes strong interferences with
important mechanisms for the prevention of simultaneous rival queen emergence.

Collective animal behaviour is a subdivision of social behaviour focussed on the coordination and emergent prop-
erties of large groups of animals achieved through group-wide transfer of information, group decision-making,
group locomotion and synchronization of activities. Many forms of collective behaviour can be observed across
multiple animal groups: murmurations of European starlings (Sturnus vulgaris)1, twisting and turning in com-
plete unison; schools of Atlantic chub mackerel (Scomber colias), that split and reform to confuse predators2;
the thousands of wildebeest that migrate across the plains of East Africa in search of better grazing3; and, most
relevant to this paper, the swarms of honeybees in search of a new home that settle on tree branches and debate
on the location of their future nest-site4.
In basic terms, swarming in honeybee colonies is a system of reproduction5. This process comprises of a
complex array of collective behaviours that have evolved to increase the probability of reproductive success: the
splitting of one colony into several new viable ones. Particularly in Apis mellifera adapted to temperate climates,
the swarming season extends across the late spring and early summer months6. In this time, a colony’s popula-
tion will expand until it is too large for its maternal nest, at which point the colony will proceed to divide itself
through the swarming process5. Seeley et al.5 states that ‘Mature colonies have a natural urge to swarm each year
unless weakened by disease or mismanagement‘. In the first swarming instance of the season, known as the pri-
mary swarm, the old queen and approximately half the worker population (of all ages) leave the parental colony
to establish a new colony, leaving behind a series of virgin daughter queens (gynes) encapsulated in the cells in
which they developed, and the remaining workers, to perpetuate the old colony. Additionally, it is common that
one or more afterswarms will ensue with a large proportion of bees leaving with virgin queens after they emerge
from their cells. The swarming bees will quickly exit the hive en masse and retain a cloud formation for around
twenty minutes until they amalgamate in a beard-like cluster at a temporary site (usually a nearby tree branch)
where they decide on their future nest-site7.
Amongst the vast array of signals, cues and behaviours, important vibro-acoustic information is conveyed
within the colony throughout some of the swarming season. This is particularly obvious from the piping of virgin

1
Nottingham Trent University, School of Science and Technology, Clifton Lane, Nottingham, NG11 8NS, United
Kingdom. 2l’Institut National de Recherche en Agriculture, Alimentation et Environnement (INRAE), UR 406 Abeilles
et Environnement, Domaine Saint-Paul, 84914, Avignon, France. 3Centre Apicole de Recherche et d’Information,
CARI, 4, Place Croix du Sud, B-1348, Louvain-La-Neuve, Belgium. ✉e-mail: [email protected]

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queens. Queen rearing is thought to be the stimulus for engorgement prior to swarming6. Once the mated queen
leaves with the primary swarm, the first virgin queen will be released from her cell between a few hours and
up to ten days after, depending on her stage in development when the primary swarm occurred6. Following
hatching, the newly emerged virgin queen will produce a type of piping known as “tooting” or “toots”, which
is thought to announce her arrival to the colony6. During each tooting sequence, the queen presses her thorax
onto the comb and vibrates her wing muscles in the folded position8. Typically, the emerged queen produces
several one-second-long pipes that are each immediately followed by several bursts of less than half a second9
with a fundamental frequency that increases with the age of queens, ranging from 200 to 550 Hz10, and is usually
observed around 400 Hz. Mature virgin queens still confined within their queen cells respond to this tooting
with their own distinct piping sound, referred to as “quacking” or “quacks”, which probably alerts the emerged
queen and the workers of their presence6. Quacking is a sequence of numerous short pulses that each typically
last less than 0.2 seconds11 at a lower fundamental frequency compared to tooting, of around 350 Hz. Both types
of queen piping are broadcast within the hive as vibrations through the honeycomb10. The function of the tooting
was examined by Grooters12 who demonstrated that a single replay of tooting inhibited the emergence of mature
encapsulated gynes engaged in chewing their way out of their cell, delaying this behaviour for several hours.
This then enabled the worker bees to reseal the cells to obstruct their emergence for up to several days12. Due to
limitations in our ability to observe honeybees under natural conditions, all such experiments to date have been
conducted under controlled conditions. Whilst this gives great insight into the proposed functions of the signals,
our long-term, non-invasive measurements provide a more ethological method for the in-situ monitoring of
honeybee hives, providing valuable continuous recordings of queen pipes occurring under natural conditions.
Most of the vibrational and acoustic information emanating from beehives is a by-product of the bees’ activ-
ities13,14 and not an outcome of direct communication signals, such as the whooping15 and the DVAV signal16,
however, this does not prevent possible high biological significance. Experienced beekeepers sometimes claim the
ability to identify certain conditions within a hive, solely by listening to its hum. In fact, the concept that swarming
can be predicted by acoustic measurements can be traced back as early as Aristotle, who wrote: “When the flight
of a swarm is imminent, a monotonous and quite peculiar sound made by all the bees is heard for several days”17. As
such, acoustic signals have been targeted in some studies for honeybee swarm prediction. In the mid twentieth
century, Woods18 obtained the sound spectrum of three colonies of healthy bees using a standard wave analyser.
He showed modulation of a 250 Hz component that he termed “the warble” in relation to the status of the colony,
particularly during the preparation for swarming. By placing microphones on the top of the brood frames within
a beehive, Ferarri et al.19 were also able to show some evidence of frequency shifts during the swarming process.
Similarly, using microphones placed close to the brood chamber, Vancata20 suggested that the intensity of the
spectrum across a frequency range of 200–500 Hz changes within the 21 days leading up to the primary swarm.
However, due to the nature of the equipment, microphone technology tends to only provide crude short-term
data collection (e.g. Woods18). In a recent paper by Zgank21, an Internet of Things based bee activity acoustic clas-
sification system is presented, with acoustic training data being collected from the Open Source Beehives Project.
However, in contrast to this paper, which aims to predict whether a honeybee colony is preparing to swarm or not,
their work focused only on identifying the best machine learning algorithm for acoustic classification between
the two states, preparing and not preparing. Owing to the use of training data originating from only one hive,
this categorisation would not be suitable for application across multiple hives21. Outside of the focus of swarming,
acoustic measurements have also been explored to make assessments into the general health status of honeybee
colonies. Robles-Guerrero et al.22 compared the sounds emanating from a healthy colony to those of a colony that
had been artificially made queenless, and Qandaur et al.23 compared healthy and Varroa-infested hives. In both
cases, preliminary results demonstrate some ability to computationally discriminate between colony conditions
based on vibro-acoustic information, but always only for a limited number of colonies.
Previous work24 focused on swarming prediction have also employed temperature sensors placed above the
brood body within a beehive to predict specific annual phases in honeybee colony development, with sugges-
tions that substantial prolonged temperature decreases could be the result of swarming or queen replacement.
Ferrari et al.19, alongside their acoustic measurements detailed above, also showed a reduction in hive tempera-
ture immediately prior to each swarming event measured across nine swarms issuing from three hives. However,
this difference in temperature occurred just minutes before the swarming event and would therefore not provide
a useful tool for swarming prediction. Buchmann and Thoenes25 and Meikle et al.26 also showed that the weight of
the hive measured with electronic scales can reveal weight changes in colonies in relation to swarming. However,
the subsequent rapid weight change can only reveal if and when the colony has already swarmed; it gives no indi-
cation as to its swarming preparations.
The above methods, as well as our own technique, contribute to “precision apiculture”, which is defined as
any honeybee management strategy that involves the monitoring of individual colony parameters (such as: tem-
perature, humidity, sound, vibration and weight) to minimise the beekeeper’s expended resources (such as: time
and money) and maximize the hive’s productivity27. With respect to the swarming process, if a system were in
place that could identify a colony’s intent to swarm, a beekeeper would benefit from being alerted to those hives
preparing to swarm and would prioritise the appropriate swarm management procedures to those hives, reducing
the need and burden for every colony to be regularly inspected, often unnecessarily. In line with Bencsik et al.13,
this study will, therefore, focus on two strategies that utilise continuous vibrational information taken from col-
onies preparing and not preparing to swarm in order to predict the swarming process in honeybee colonies. The
aim of this work is to explore whether there are features found in the honeycomb vibrational spectra that are
specific to colonies’ preparations to produce a primary swarm. These features are then used to raise an alarm, as
early as possible prior to the event. This technique encompasses the use of accelerometers placed directly in the
centre of honeybee hives and continuously collects the average spectra of vibration at three minutes intervals (see
Methods). A reliable signature will be one which will detect a high proportion of swarms. A more reliable one

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will work for swarm events that have not contributed to the numerical search (training stage) originally required
to clarify the signature. The best vibrational signature will be one that will work from season to season, and for
different apiaries. Usually the primary swarm is not preceded by any piping, but nevertheless we have also endeav-
oured, in this study, to carefully examine the pipes logged within our continuous vibrational datasets, as it seems
to strongly support the theory that tooting and quacking are produced to inform the worker bees about the need
to release or keep captive un-emerged virgin queens.

Results
Prediction of swarming. Two strategies are presented here that both use averaged vibrational spectra for
the prediction of swarming: the first (Fig. 1), utilises discriminant functions created from instantaneous (dismiss-
ing the influence of any past data) averaged vibrational spectra (Fig. 2), which only performed well when restric-
tively applied over the swarming season (Fig. 3), and a second (Fig. 4) that uses three dimensional Fourier analysis
(3DFT) to account for the evolution of the previous ten days’ spectra. For both strategies, in the first instance, a
numerical search was required to define those Discriminant Functions (Fig. 2 for instantaneous functions and
Fig. 5 for spectral evolution functions) that best enable the classification of vibrational spectra as being that of a
colony in a state of intending to swarm or not, as described for each strategy in Figs. 1 and 4. In-line with Ramsey
et al.15,16, this numerical search consists of successively applying a data reduction exercise (Principal Component
Analysis, PCA) followed by a supervised clustering exercise (Discriminant Function Analysis, DFA) on the selec-
tion of PCA scores. To best enable this to identify those measurements that correspond to (i) colonies preparing
to swarm and (ii) colonies not preparing to swarm, specific measurements that exhibited the most generic fea-
tures for these two conditions (see Bisele, et al.28) were supplied as ‘training data’. For more information on how
the two conditions were identified and discriminated, refer to the Methods.

Strategy 1 – Discrimination of instantaneous spectra. From a possible 253 one-hour-long averaged

spectra from colonies preparing to swarm (PTS) and 1754 from non-swarming (NS) colonies (see Methods),
parametric optimisation (see Methods) revealed that the best performance from the software was obtained when
using a training database size of 24 PTS and 32 NS 3-minute averaged logarithmic spectra. These are further
averaged between 3:00 and 4:00 AM over the 13 days leading up to the primary swarm and were cropped over
a bandwidth of 18 to 800 Hz, with 40 percent of the PCA scores being projected onto three DF scores (Fig. 2).
These parameters yielded a 6.3% classification error (percentage of misclassified averaged spectra over the entire
collection of available data). The three discriminant spectra in Fig. 2, resulting from our numerical search, have
very different spectral features, and they reveal the highest contribution to originate at around 20 Hz, with DF3
exhibiting the majority of its overall information at 24 Hz and its harmonic frequencies. Vibrations above 200 Hz
appear to contribute modestly to the discrimination, and those towards the highest frequencies up to 800 Hz,
even less so.
In Fig. 3, two specific colonies’ instantaneous alarm is computed across the entirety of their respective data-
sets, showcasing the outcome of the algorithm for a colony that swarmed (Fig. 3a) and another that did not
(Fig. 3b). For the extensive collection of alarm plots pertaining to all colonies see Figs. S1–S14 (non-swarming)
and Figs. S15–S32 (swarming) of the supplementary material.
In Fig. 3a, we see a representative example that shows that our discriminant functions (Fig. 2) are successful
in discriminating between the change of preparation status between a colony that is aiming to swarm and the
comparatively steady-state of the preparation status of a colony that is not aiming to swarm (Fig. 3b). Affected by
the spectral variations induced by foraging activities, the alarm is not triggered for data leading up to the primary
swarm that were collected between 9:00 AM and 4:00 PM. The spectra at these peak foraging times must there-
fore resemble that determined by our software as a non-swarming spectrum. Similarly, the hourly alarm is also
triggered during foraging hours later in the season, as shown by occasional purple pixels in July and August. By
averaging the night time alarm between 12:00 AM and 5:00 AM, giving a daily assessment of a colony’s prepa-
ration to swarm, we show that our algorithm, for this specific colony, is able to determine that the colony was
going to swarm one month before the exit of the primary swarm. We also show that for a specific, representative
colony that did not swarm, the alarm was never triggered (Fig. 3b) either on an hourly or daily basis. Alarms
displayed across Supplementary Figs. S15 to S22 show that the placement of the accelerometer across the frames
of the broodbox is nearly irrelevant for the prediction of swarming, with all eight frames monitored across the
season for this specific colony successfully producing a swarming alarm several days before the swarming event.
This suggests that the accelerometers embedded in the honeycomb pick up the entire colony’s overall buzzing,
irrespective of the frame on which they were affixed.
For swarming colonies (Supplementary Figs. S15–S32), based on the mean (red line) of the night time alarm,
the alarm was successfully triggered in 15 out of 18 datasets before the exit of the primary swarm. Moreover, the
alarm was triggered on average twenty-two days before the exit of the primary swarm. For the non-swarming
colonies, no false positives are found across the collection of datasets (Supplementary Figs. S1–S14), based on the
mean night-time alarm, giving a 91% success rate in the software’s ability to discriminate between our swarming
and non-swarming colonies. Note that the black sections seen within the hourly alarm plots across the 2014
French colonies between the 17th and 24th March, the 17th and 24th August and the 14th and 27th November are the
result of missing data in the datasets, due to power cuts. This caused the mean alarm to drop below the threshold,
as their values are strictly zero; those values were not considered as false detections.
Despite all our efforts, false positives are still triggered occasionally on an hourly basis, and this becomes
exacerbated when the time duration of the season under scrutiny is extended to the rest of the summer. We
also explored whether our algorithm was lacking the required sensitivity, or whether the data were genuinely
lacking specificity to the preparation of swarming. To this end, a simple cross-correlation exercise was used to
compare the similarity between individual swarming and non-swarming spectra which make up each training

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Figure 1. The instantaneous alarm procedure. The images in Fig. 1 are solely for illustrative purposes. This
figure depicts the processing for the detection of swarming in the post-training phase. Each hour-long averaged
spectrum is compared, using the cross-correlation product (CCP), to a pair of DF curves, as determined by the
DFA algorithm (see Fig. 2), providing the co-ordinates for a single point in the DF space (green ‘+’ sign). The
Euclidean distance to the swarming and non-swarming centroids (the central point of all swarming and non-
swarming spectra, used in the training phase, respectively shown as a red dot with black outline and green dot
with blue outline) are calculated. The ratio of these distances provides our “criterion value”, and is compared to
an optimised threshold value (see Methods) to determine if it is the spectrum of a colony intending to swarm
or not, i.e. if the point in DF space lies within the upper left “swarming area” bordered by the yellow line. In
this figure, for ease of representation, we give an example using two discriminant functions, however, in this
study, three discriminant functions were used. Plots used in the creation of this figure were generated using the
MATLAB (R2018b) software (uk.mathworks.com).

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Figure 2. The optimised instantaneous discrimination spectra (DF1, DF2 and DF3) as defined by the DFA
algorithm. Frequency along the X-axis is cropped between 18 and 800 Hz; the Y-axis shows the logarithm of
acceleration. High (positive or negative) acceleration values indicate spectral areas of particular importance to
swarming prediction, whilst low values, approaching zero, correspond to vibrational frequencies which must be
dismissed. This figure was created using the MATLAB (R2018b) software (uk.mathworks.com).

Figure 3. Time course of the swarming alert for a specific colony that (a) swarmed and (b) did not swarm,
using the DF spectra in Fig. 2. The instantaneous alarm (computed without any contribution coming from past
data) for these two colonies was monitored from the 13th March until 1st December 2014. The colour coding has
been split to clearly convey two very different pieces of information. Greyscale tones correspond to the alarm
values above the threshold (non-swarming state) and the remaining colour emphasises the alarm values below
the threshold (swarming state), from pink to blue as the criterion approaches the swarming centroid. Green
rectangles show the occurrence of three swarms within this dataset, with the earliest one being the primary
swarm. Superimposed is another, red curve, with its own Y-axis shown on the extreme right, showing the
average of the previous night’s alarm values taken between midnight and 5:00 AM. The yellow line displays the
alarm threshold. Note that the black sections seen within the hourly alarm plots are the result of missing data,
due to power cuts. This figure was created using the MATLAB (R2018b) software (uk.mathworks.com).

database. This revealed that night-time instantaneous spectra taken from a colony preparing to swarm can, in
some instances, be virtually identical to that taken outside of the swarming season, or from a colony not preparing
to swarm (Supplementary Fig. S33). This lack of specificity of these particular data therefore suggests that there is
no need to seek for more advanced algorithms to discriminate the required categories. We therefore moved on to
the exploration of a new feature, one accounting for the recent history of the evolution of the spectra, effectively
removing the flaw of finding identical spectra in the PTS and NS clusters.

Strategy 2 – Discrimination between Spectral Evolutions. Parametric optimisation explored a col-

lection of 126 swarming and 1512 non-swarming 3DFT (three dimensional Fourier transform) spectra, and used
an optimised training database based on 20 swarming and 21 non-swarming spectra, obtained between midnight

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Figure 4. The spectral evolution alarm procedure. The images here are solely for illustrative purposes. This
figure depicts the process for the detection of swarming in the post-training phase. In order to account for the
history of the vibrational spectra, the FFT (Fast Fourier Transform) is performed on the time course of the
magnitude of all the uploaded spectral frequencies for each day between midnight and 5:00 AM, yielding 2DFT
(two dimensional Fourier Transform) images (second line). The FFT is then further calculated over each pixel of
the series of 2DFTs found in the preceding days (third line). Each 3DFT (three dimensional Fourier Transform)
is compared to a pair of Discriminant Functions, as determined by the DFA algorithm (Fig. 5), using the cross-
correlation product (CCP), the values of which determine the co-ordinates for a single point in the DF space.
The remainder of the procedure is identical to that described in Fig. 1. In this figure, for ease of representation,
we give an example using two discriminant functions, however, in this study, three discriminant functions
were used. Plots used in the creation of this figure were generated using the MATLAB (R2018b) software (uk.
mathworks.com).

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Figure 5. The three discrimination spectra for the spectral evolution algorithm. The pixel colour indicates
the negative (blue), positive (red) or zero (green) contribution the user must give to a particular part of the
vibrational 3DFT in order to get the first (top), second (middle) or third (bottom) DF score. The frequency
components of the FFT of the 2DFTs are shown stacked from left (0 repetitions per week) to right (3.5
repetitions per week) for ease of viewing. This figure was created using the MATLAB (R2018b) software (uk.
mathworks.com).

and 5:00 AM, incorporating the previous 10 days of history leading up to the primary swarm, and cropped over
a bandwidth of 25 to 800 Hz, with 40 percent of the PCA scores being projected onto three DF scores (see Fig. 4
for more information). This resulted in the best performance from the software (as displayed in Fig. 6). These
parameters yielded a 1.92% classification error between the 1638 (126 swarming and 1512 non-swarming) data
points used in the clustering exercise. The DF spectra in Fig. 5 exhibit features that are very different from those
seen in Strategy 1, with most variation observed below 700 Hz.
Swarming alarm values based on the ten-day evolution of spectra for two individual colonies are computed
across the entirety of their respective datasets, showcasing a colony that did (Fig. 6a) and did not (Fig. 6b)
swarm. For the extensive collection of alarm plots pertaining to all non-swarming and swarming colonies see
Supplementary Figs. S34–S47 (non-swarming) and Supplementary Figs. S48–S64 (swarming).
In Fig. 6, we demonstrate that our discriminant functions incorporating the 10-day spectral evolution (Fig. 5)
are successful in generating mean criterion values that discriminate between a colony that is and is not preparing
to swarm, with the alarm triggered and sustained 16 days before the exit of the primary swarm in the example. In
addition, the daily mean midnight to 5:00 AM criterion value did not trigger the alarm after the exit of the final
afterswarm (except perhaps temporarily on the 10th May). Due to the variation induced by foraging activities, as
seen for the instantaneous method in Fig. 3, the alarm is not triggered for spectral data from between the hours
of 6:00 AM and 4:00 PM leading up to the primary swarm. The spectra at this prominent foraging time must
therefore, again, more closely resemble those determined by our software as non-swarming spectra. In addition,
the hourly alarm is sporadically triggered during foraging hours later in the season, e.g. 9th August and 15th
September, at a similar rate to that of Fig. 3. These false positives can be effectively dealt with by averaging the
resulting alarm from midnight to 5:00 AM. Alternatively, these alarms may be meaningful, as in north-eastern
North America scientists have shown that in addition to a major period of swarming in May/June, a minor period
of swarming also exists in September.
We also show a representative example of a colony that did not swarm (Fig. 6b) where the alarm is never
triggered, based on the alarm averaged between midnight and 5:00 AM. Note that the hourly alarm for the
non-swarming colony is only ‘mildly’ triggered a few times around 8:00 PM between the 1st and 4th November, as
alarm values are very close to that of the threshold, performing similarly to that of the instantaneous spectrum
alarm in Fig. 3b.

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Figure 6. Swarming alarm based on the 10-day evolution of spectra for a colony that (a) swarmed, situated
at Nottingham Trent University and (b) did not swarm, situated at INRA, France. Each colony was monitored
across the active season in 2014. The colour coding is identical to that of Fig. 3. Green rectangles display the
timings of three swarms that occurred within this dataset. Super imposed is another graph with the y-axis (at
right) being the daily mean criterion value computed from data between midnight and 5:00 AM; the yellow line
displays the alarm threshold. This figure was created using the MATLAB (R2018b) software (uk.mathworks.
com).

For the non-swarming colonies (Supplementary Figs. S34 to S47), based on the mean midnight to 5:00 AM
alarm, only two colonies produce false positives within the midnight to 5:00 AM average alarm, across the entire
collection of datasets. These false alarms occurred within the 2014 French colony 5 dataset between the 28th
April and 3rd May 2014 (Supplementary Fig. S37), and the 2014 French colony 22 between the 10th and 20th July
(Supplementary Fig. S47). For swarming colonies (Supplementary Figs. S48–S64), based on the mean of the night
time alarm, in 13 out of 18 datasets, the alarm was successfully triggered before the exit of the primary swarm,
but it was not successful in predicting swarms for the 2014 French colonies 10 (Supplementary Fig. S58) and 13
(Supplementary Fig. S59), as well as 2015 French colonies 15 (Supplementary Fig. S63), and 19 (Supplementary
Fig. S64). This gives an overall success rate of 80% in the software’s ability to discriminate between our swarming
and non-swarming colonies, with an average of 10 ± 2 days (n = 11) warning (calculated from those colonies that
were correctly classified as swarming colonies), when using the mean midnight to 5:00 AM mean criterion values
to assess the swarming preparation of honeybees. As with the previous method using the instantaneous spectra,
Fig. 3a and Supplementary Figs. S48 to S54 reveal that 8/8 frames monitored across the UK colony 1 hive suc-
cessfully triggered a swarming alarm several days preceding the swarming event, suggesting again that the lateral
placement of the accelerometer does not affect the prediction of swarming using this technology.

Comparison between the two strategies. In Table 1, we see a direct quantitative comparison between
the performances of the two strategies for the prediction of swarming using averaged vibrational spectra. With
22 days compared to 10.1 days, we can see that strategy 1, using instantaneous spectra, provides a much longer
swarm warning than strategy 2, using the spectral evolution. When using the average daily criterion values, only
3 colonies were wrongly classified from the collection of 32 when using strategy 1, compared to the 7 misclassified
by strategy 2. ‘Raw alarm false positives’ refers to the number of colonies that have hourly alarm triggers (criterion
value falls below established swarming threshold) when considered not in a state of preparation to swarm, i.e.
after the last queen pipe or from a non-swarming colony. We see here that strategy 1 and 2 performed equally,
with all colonies exhibiting at least some false positives on an hourly level. ‘Raw alarm false negatives’ refers to the
number of colonies where, on an hourly basis, no alarm trigger occurred when the colony was considered to be
in a state of preparation to swarm, i.e. in the two weeks leading up to the primary swarm. In this respect, strategy
1 again outperformed strategy two, with only two swarming colonies lacking an hourly alarm trigger compared
to the three of strategy 2. It is clear from this comparison that strategy 1 was much more effective than strategy
2 and that the calculation of the early morning mean daily criterion value is an essential tool for averaging out
hourly false positives.

Queen Pipes
Toot and quack signals characterisation. We now proceed to carefully examine the queen pipes residing
within our measurements. As raw audio recordings were also available for the Jarnioux 2015 dataset, we com-
bined them with the daily spectrograms of the 3-minute averaged spectra to draw further conclusions regarding
the occurrence of queen toots and quacks in each of our datasets (Supplementary Figs. S65 to 72). In Fig. 7, a daily
spectrogram of vibrations can be seen for the Jarnioux dataset of 30th April 2015, 9 days after the primary swarm,
for both the central (Fig. 7a) and peripheral (Fig. 7b) accelerometers. Within the data of this particular day, we
see the general buzzing of the bees that occurs at 125 Hz and 250 Hz, its upper harmonic. In addition to these two

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Figure 7. Continuous raw accelerometer data, averaged over three minute intervals, for frequencies between
0 and 1000 Hz, recorded from a single colony in Jarnioux, France, on 30th April 2015. Data are shown for the
accelerometers situated at the centre (a) and at the periphery (b) of the frame. Data shown were recorded on
the 30 April 2015. Colour coding represents logarithmic spectral amplitude from low (blue) to high (red) in
arbitrary units. Highlighted in the black rectangle is the short sample of data that is represented in Fig. 8. This
figure was created using the MATLAB (R2018b) software (uk.mathworks.com).

Figure 8. Spectrogram of raw spectra containing the toots and quacks found in the section highlighted in
Fig. 7. Data are shown for the accelerometers situated at the centre (Top, a) and at the periphery (Bottom, b) of
the frame. Pixel intensity represents logarithmic spectral amplitude from low (blue) to high (red) in arbitrary
units. Highlighted in the red rectangle is the small sample of data further analysed in Fig. 9. This figure was
created using the MATLAB (R2018b) software (uk.mathworks.com).

highly reliable continuous traces, two sets of intermittent peaks can be seen between 300 and 350 Hz that diverge
at around 5:00 AM, with one set exceeding 400 Hz whilst the other remains at 300 Hz. A small section of these
data from 8:00 AM, located within the black rectangle, was extracted, analysed separately and displayed in Fig. 8.
Through critical listening and visual inspection, we ascertained that the higher frequency band at around 400 Hz
specifically pertains to queen toots and the lower frequency vibrations at around 300 Hz correspond to the quack-
ing of encapsulated gynes (Fig. 8). The audio sample of these virgin queen vibration signals presented in Fig. 8
can be found in Audio S1. When we further home onto these (Fig. 8), we can see the differences in the spectral
properties of the two types of queen pipes. In the example showcased in Figs. 8 and 9, queen tooting consists of a
1-second-long narrowband pipe followed by a train of multiple (eight in this case) 0.2-second-long pulses of the
same frequency. In contrast, the quacking by cell-captive mature gynes held in their cells by worker bees is com-
prised of a series of usually many 0.2-second-long pulses that occur around 300 Hz. These are much more severely
frequency-modulated and resemble the U-shaped figure of the train of pulses that make up the last few pulses of
the queen toot, only at a lower frequency. In particular to the signal showcased here, seen in Fig. 9, tooting occurs
in intermittent 5-second-long signals, whereas quacking persists uninterrupted for over 20 seconds. It can also

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Figure 9. Spectrogram of raw spectra containing the toots and quacks found in the section highlighted in
Fig. 8. Data shown are for the accelerometers situated at the centre (Top, a) and at the periphery (Bottom, b) of
the frame. Colour coding represents logarithmic spectral amplitude from low (blue) to high (red) in arbitrary
units. This figure was created using the MATLAB (R2018b) software (uk.mathworks.com).

Quantitative criterion Strategy 1 – Instantaneous Spectra Strategy 2 – Spectral Evolution

Averaged predictive warning 22 days 10.1 days
Averaged alarm errors
(number of colonies which
produced false negatives
(swarming) or false positive
(non-swarming) when using
the mean criterion value) Swarming Colonies Non-Swarming Colonies Swarming Colonies Non-Swarming Colonies
3/18 0/14 5/18 2/14
Raw alarm false positives
(number of colonies with hourly
alarm triggers when considered Swarming Colonies Non-Swarming Colonies Swarming Colonies Non-Swarming Colonies
not in a state intending to
swarm)
18/18 14/14 18/18 14/14
Raw alarm false negatives
(number of colonies with
no hourly alarm triggers Swarming Colonies Non-Swarming Colonies Swarming Colonies Non-Swarming Colonies
when considered in a state of
intending to swarm)
2/18 N/A 3/18 N/A

Table 1. Quantitative comparison of the performance evidence for each swarm prediction strategy discussed in
this paper.

be seen that the toots and quacks are much more prominent on the spectrograms pertaining to the peripheral
accelerometer compared to that of the central one. This is due to the location upon the comb in which each type
of signal was produced. During the swarming process, queen cells are usually built at the periphery of the frame
(as opposed to the centre during emergency queen rearing), and queens tend to toot nearby (Grooters, 1987). As
a result, both piping signals have a much greater magnitude on the peripheral accelerometer as compared to the
central one (Figs. 7, 8 and 9).

Chronology of toots and quacks in disturbed and undisturbed colonies. In Fig. 10, we see data
from a colony (French Colony 14, 2014) that originally had a clipped queen, and whose hive was opened several
times during the swarming season. Faint, scarce and irregular virgin queen tooting started on the 15th April 2014
two hours after a large colony level disturbance caused by planned hive maintenance that took place. It suggests
that the event most probably resulted in the early release of an encapsulated gyne. It is difficult to see the tooting
on the spectrogram in Fig. 10 due to the low density of its occurrence, however in Supplementary Fig. S73, we
depict the hour pertaining to the beginning of tooting on the 15th April. This shows a high amplitude broadband
spectrum, synonymous to that obtained when a hive is opened, that occurred at midday on the 15th April. Piping
then started two hours later, as highlighted in Supplementary Fig. S73. A swarm was attempted and failed the
next day on the 16th April, likely due to the clipped wings of the original queen. The swarm is identified by a high
amplitude broadband vibrational peak, especially at ultra-low frequencies around 60 Hz, twice lower than that of
the 125 Hz buzzing, which occurs at the time of the lift-off, with a significant drop in signal amplitude thereafter

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Figure 10. Spectrogram image of 3-minute averaged spectra extracted from the 2014 French colony 14 dataset,
showcasing the occurrences of queen piping recorded within a colony that contained a clipped queen and
was disturbed throughout its swarming activities between 15th April and 2nd May 2014. Midnight of each day
begins at each date label on the horizontal axis. Pixel intensity represents logarithmic spectral amplitude from
low (blue) to high (red) in arbitrary units. This figure was created using the MATLAB (R2018b) software (uk.
mathworks.com).

Figure 11. Spectrogram image of 3-minute averaged spectra extracted from the 2015 French colony 22 dataset,
representing the occurrences of queen piping recorded within a colony with an original queen that was left
unclipped and was left undisturbed throughout its swarming activities between 13th and 22nd April 2015.
Midnight of each day begins at each date label on the horizontal axis. Pixel intensity represents logarithmic
spectral amplitude from low (blue) to high (red) in arbitrary units. This figure was created using the MATLAB
(R2018b) software (uk.mathworks.com).

(see Supplementary Fig. S74). When a swarm fails and returns to the hive, a large peak occurs at a fundamental
frequency of around 150 Hz, just after the swarming event, as the bees re-enter which causes a buzzing vibrational
amplitude increase rather than the expected decrease (Supplementary Fig. S74). The hive was opened again on
the 17th April. Following this, the 400 Hz virgin queen tooting can be seen with greater intensity than previously
observed. However, the toots appear much scarcer and sporadically distributed, (24 hours between consecutive
toots) than would normally be expected from a newly emerged tooting queen (see the 17th April onwards in
Fig. 11). This is perhaps another consequence of a queen having been released early due to human intervention.
Another failed swarming attempt occurred on the 24th April. A successful swarm finally ensued on the 26th April,
which resulted in the large decrease of the buzzing vibrational signal and the abrupt cessation of queen tooting,
strongly suggesting that a virgin queen left with the swarm. This phenomenon is also seen for 2014 French colo-
nies 1, in Supplementary Figs. S65 and S6, in Supplementary Fig. S66, where tooting can be observed before the
exit of the first successful swarm. As seen in Fig. 10, the hive was opened for a third time on 27th April, resulting in
new queen tooting a few hours later. Quacking then ensued after six hours. Tooting and quacking both increased
in frequency from 400 Hz to 500 Hz, and from 300 Hz to 400 Hz, respectively. Both tooting and quacking then
terminated upon the exit of the final afterswarm, suggesting that an emerged virgin queen left with the swarm and
an encapsulated gyne emerged and stayed within the colony.
In Fig. 11, the data for the swarming events of French Colony 22 in the 2015 season are shown. The original
queen was unclipped, and this particular colony was left undisturbed throughout its swarming activities. As
expected, no piping can be seen before the primary swarm that issued at 2:00 PM on the 13th April. Frequent
queen tooting started four days later on the 17th April and increased in signal frequency from 300 Hz to 450 Hz
in the two days leading up to the first afterswarm. Six hours after the start of tooting, quacking can be observed
to increase from 280 Hz up to 350 Hz, which then holds stable until cessation at the second afterswarm. In all
cases (Supplementary Figs. S65–S72), tooting always preceded quacking. After the first after swarm, there was a
three-hour hiatus in queen tooting for this particular colony, whilst the quacking persists, and there appears to
be a reduction in the occurrence of quacking, a phenomenon also seen across the other datasets (Supplementary
Figs. S65–S72). This suggests that the emerged virgin queen left with the swarm and a previously encapsulated

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gyne emerged. Both tooting and quacking then persisted until the final swarm after which, like all other colonies
(Supplementary Figs. S65–S72), they are no longer apparent in the dataset.

Discussion
In this work, we disclose a non-invasive method for the prediction and the monitoring of the swarming process
within honeybee colonies. As such, two machine learning algorithms, along with their respective discriminant
functions (Fig. 2 for instantaneous spectra alarm and Fig. 6 for the spectral evolution alarm) that act as generic
signatures for swarming and non-swarming classification, are presented for the prediction of swarming. These
discriminant functions are based on the overall three-minute averaged vibrational spectra recorded using accel-
erometers placed in the comb among brood cells at the heart of honeybee hives. Averaging the accelerometer data
removes the need to create exceptionally large databases containing the raw data pertaining to each hive, as seen
for Ramsey et al.15,16, and provides data relevant to the overall colony buzzing states, irrespective of the frame of
the hive in which the sensor is placed.
The primary swarm is remarkably difficult to predict from accelerometer measurements since it is not associ-
ated with any queen piping. On the contrary, secondary swarms always appear to take place after many hours of
queen pipes that are straightforward to detect as soon as they appear in the colony. The main challenge that this
work endeavoured to overcome is the prediction of primary swarm events when no obvious swarming-specific
features exist within the averaged spectrum. As such, both of our algorithms can successfully discriminate
between swarming and non-swarming colonies with a high degree of accuracy, with the analysis based on instan-
taneous spectra performing with a classification error of 6.3% within the swarming season, and the spectral
evolution analysis performing with an error of 1.91% without any restriction on the season under scrutiny. In this
paper, we also disclose an original analysis of the accelerometer trace pertaining to, and an extensive collection of,
queen toots and quacks, which allowed for us to disclose new features of queen piping in relation to other colony
events, over the entire swarming season for each hive, both for inspected and non-inspected colonies.
Provided that the user restricts the application of the instantaneous spectra method for swarming prediction
to data from the swarming season alone, this algorithm works remarkably well. Even whilst necessitating one
hour of recording each day between 3:00 and 4:00 AM, the instantaneous spectra only failed to predict three
out of eighteen primary swarms and did not exhibit a single false positive from the non-swarming colonies.
Further investigation into the false positives exhibited outside of the swarming season revealed that instances
exist throughout the swarming and non-swarming training databases where the night-time spectra taken from
colonies that are considered in a state of preparing to swarm are almost identical to spectra extracted from outside
the confines of swarming (see the example in Supplementary Fig. S33). This demonstrates the poor specificity
of instantaneous spectra to swarming prediction and encouraged us to explore an alternative history-carrying
feature in order to lift the obvious lack of specificity of instantaneous spectra. Between the three discriminant
functions generated as part of the training phase of our discrimination exercise, we also show that there is little
information above 700 Hz useful to the prediction of swarming. In fact, the largest variation, and therefore the
most important information, is shown to occur around 20 Hz, which matches remarkably well with the frequency
of the DVAV signal16. In previous work16, we have shown that our accelerometer technology is very effective for
the detection of DVAV signals, for which the generally accepted function is that of “prepare for greater activity”29
and that enhancements in this signal can be seen prior to the exit of the primary swarm. From the stronger
emphasis that our discriminant spectra demonstrate around 20 Hz compared with frequencies above 100 Hz,
we can deduce that the general buzzing of the bees alone does not give important or reliable information per-
taining to a colony’s intent to swarm. This is in direct contrast to Woods18, Ferrari et al.19 and Vancata20 who
have suggested that sound spectra above 200 Hz are modulated by honeybee activity with high specificity in the
days leading up to the primary swarm with high specificity. The 20 Hz feature we reveal to be imperative to the
prediction of the swarming process using spectral information also undermines previous authors work using
microphones18–21 because DVAV signals are a non-audible signal and microphones are incapable of detecting fre-
quencies as low as 20 Hz. An accelerometer, on the other hand, has no limitations at such low frequencies, adding
further substantiation to the use of accelerometers for monitoring honeybee hives.
The improved accuracy of the discrimination procedure based on the ten-day spectral evolution can easily
be seen in the hourly alarm plots, with far fewer false positives occurring on an hourly basis after the end of the
swarming season for the ten-day spectral evolution than that based on the instantaneous spectra. One drawback
of the spectral evolution method, however, is that at least ten days of data are required before the alarm value can
be computed and assessed, and this requires a much larger storage capacity. Additionally, the software to run
the analysis is much more computationally demanding when compared to analyses based on the instantaneous
spectra. The alarm for each method is significantly further improved if it is averaged over the hours of midnight
and 5:00 AM, for the instantaneous spectra, and 8:00 PM and 1:00 AM for the alarm based on the spectral evolu-
tion. This provided a daily averaged alarm that maintained true alarms and removed sporadic false positives (See
Fig. 3). From this, it is seen that the instantaneous spectra method outperformed that of the spectral evolution in
terms of the number of days prior to swarming that the alarm was triggered and the low number of false positives
within the non-swarming data. This indicates that, in one way or another, information regarding the ‘history’ of
the spectra is required. In the context of a commercial product aimed at aiding the beekeeper, it could be argued
that false positives that arise using either algorithm after the end of the swarming season for colonies that did
swarm are irrelevant, because a swarming detector would be unnecessary after the departure of the primary
swarm or after the swarming season has come to an end (after July). Nevertheless, late (e.g. September) swarming
and absconding outside the typical swarming season are phenomena known to occasionally take place30. In future
work, it would be highly interesting, should such data be available, to see whether our alarm would success-
fully detect fall swarming events. The lack of specificity of the features found in the instantaneous spectra causes

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numerous false alarms during busy foraging times of the day. The use of early morning data is best for colony
status assessments, as successful foraging afternoons, can lead to enhanced vibrational activities until midnight.
Both strategies could perhaps be further improved through the inclusion of additional grouping clusters at the
training phase from colonies that exhibit data that account either for honeycomb density due to differing brood
levels or comb contents (e.g. honey and pollen), or that have some feature (e.g. ‘varroa infested’ or ‘not varroa
infested’) that makes their vibrational spectra differ from others. These algorithms can also be easily developed to
determine the likelihood (percentage) that a colony is going to swarm, based on the alarm values calculated from
the ratio of distances to each centroid, of the percentage likelihood that a colony is going to swarm. This would
allow the beekeepers to prioritise the hives that require the most urgent inspection.

Queen pipes. In addition to our algorithm for the prediction of primary swarms, our daily spectrograms give
new insight into the physical features of queen piping following the primary swarm. As such, this work allowed
for comparison of the waveform properties of toots and quacks across the entire swarming season obtained by our
accelerometer measurements to that of the laser vibrometer (a method unable to provide the long-term measure-
ments that accelerometers can) used by Michelsen et al.10 (described in the introduction). Our results substantiate
theirs. For tooting, we show that there is a frequency increase from 350 Hz to 500 Hz that occurs gradually as the
first gyne to emerge ages, reaching a steady state just before she exits with an afterswarm. The analysis of quacking
also showed a similar phenomenon consisting of a rise in frequency from 200 Hz up to 350 Hz over a few succes-
sive days. It was also found, through critical listening and close examination of spectra that pertained to the raw
accelerometer audio of the Jarnioux 2015 dataset, that queen tooting consists of a 1–2 second pipe followed by a
train of 0.2 second pulses of the same frequency, as described by Wenner9 and Michelsen10. In contrast, quacking
by gynes confined in queen cells consists of long series of 0.2 second pulses that occur around 350 Hz. These
results also correspond very well with the physical properties described by Kirchner11. We also found that queen
quacks exhibit much broader spectral variation around the fundamental frequency than the much narrower band
found in queen toots (Fig. 7). Owing to these spectral differences, queen toots and quacks can be discriminated by
eye in most instances, within the 3-minute average spectra (e.g. see the 5:00 AM divergence of 350–500 Hz toots
from steady 350 Hz quacking in Fig. 7), allowing for the visual detection of their natural occurrences within the
continuous vibrational datasets available to us in this study.
The Jarnioux 2015 colony (Supplementary Fig. S72) and the 2015 colony 22 (Fig. 11) were left undisturbed
with non-clipped queens during their swarming processes. As can be seen in the data and corroborated by the
raw accelerometer audio, queen tooting started four to seven days following the exit of the primary swarm, with
quacking ensuing after the initiation of tooting. This duet of tooting and quacking persisted until the exit of an
afterswarm when tooting ceased for a few hours before being logged again, strongly suggesting that the next
gyne had been released from her cell. This sequence of events repeats itself until the exit of the final afterswarm,
at which time there is not a single gyne remaining in her cell. In some instances, queen tooting continues for
several days after the exit of the final swarm. As such, our ethological approach provides strong evidence in sup-
port of the theory of the queen piping being a colony-level communication to aid the worker population in the
co-ordination of the release of queens by conveying information about how many queens are free and encapsu-
lated to avoid competition between them (as described in Winston6). In the undisturbed colonies, if it were not
for the worker bees actively keeping the encapsulated gynes captive, then two or more queens would be observed
tooting contemporaneously (this is never heard in the raw accelerometer recording). The timings of our logged
toots and quacks are very much in line with the work of Grooters12, further substantiating their conclusions that
queen tooting was an important mechanism for the prevention of rival queen emergence. Our results also further
substantiate the well-accepted idea that only mobile queens produce tooting signals, since the issue of secondary
swarms consistently resulted in the abrupt cessation of tooting, even if only for a few hours.
An exciting further experiment would be to drive artificially generated toot signals into the hive of a colony
immediately following the exit of the first afterswarm to reveal if workers bees extend the duration in which gynes
remain captive. In a study by Simpson and Greenwood31 artificial 650 Hz pulsed vibrations were generated to
mimic queen pipes in hives of colonies containing a single mobile gyne and concluded that this encouraged them
to swarm. The conditions under which this experiment was conducted does not allow for us to explore the effect
that this artificial piping could have had on worker bees maintaining encapsulated gynes, and possibly suggests
that tooting sensed by a mobile gyne, indicating the presence of a second one, in the absence of quacks, is more
likely to result in her exit with a swarm than in the absence of any pipe.
In colonies that were disturbed several times during their swarming procedures or in colonies contain-
ing queens with clipped wings, an entirely different set of phenomena were observed. Hive 1 (Supplementary
Fig. S65), 10 (Supplementary Fig. S67) and 13 (Supplementary Fig. S68) from France in 2014, for example, had
several failed swarming attempts, which can be seen as a large high-magnitude buzzing trace that appears after the
swarm as the bees unexpectedly re-entered their hive, as opposed to the expected and otherwise observed loss of
vibrational signal that would normally result from a large amount of bees exiting the hive. This transpired because
the queen with clipped wings was unable to fly with the swarm. These colonies were also opened several times
during the early stages of the swarming process. This led to queen pipes occurring before the first swarm that
ceased upon the successful exit of the swarm. This suggests two possibilities 1) there were two (or more) emerged
queens (the old queen and a newly emerged virgin gyne) present in the hive at the same time, due to inspection
disturbance causing the early release of a virgin queen, and the primary swarm left with the additional virgin
queen, or 2) it was the original mated queen that was piping during this time, who then left with the primary
swarm, indicating that mated queens, one or more years of age can also pipe32. However, the ages of the queens
were not known. In colonies with a queen that cannot fly, a range of phenomena can take place (see footnote).
Footnote for this page only: personal communication from Professor Gard Otis, who observed 4 colonies that
swarmed with virgin queens without having issued a prime swarm with the mated queen. He referred to those

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swarms as “afterswarms” because they had virgin queens. In two instances the mated queens were unable to fly
with the swarm, and they disappeared once a virgin queen emerged from her cell; those colonies proceeded to
issue afterswarms with virgin queens. However, in one case, the mated queen remained in the hive as it swarmed
with her present colony; two afterswarms with virgin queens issued, after which the original mated queen once
again became the egg-laying queen of the colony. She later did leave with a prime swarm in the subsequent
swarming cycle (see Otis33).
Our method and results strongly suggest that fundamentally important colony disturbance can be brought
by beekeeping activities during the rearing of virgin queens, which is a time when more frequent beekeeping
inspections are recommended. In colonies left undisturbed with non-clipped queens, our methods have pre-
served what can be considered the natural pre- and post-swarming behaviour of honeybees. We have demon-
strated that such natural behaviour can be lost through invasive inspections, resulting in the early release of a
virgin queen, so that more than one mobile queen is present within the hive. This is not a natural situation for the
colony, which otherwise delivers an orderly release of gynes in relation to swarming events6. This also supports
the suggestion that tooting acts to aid in the prevention of the simultaneous emergence of rival queens, as during
physical inspections by beekeepers, tooting would be interrupted on each frame as it is removed for inspection,
allowing the opportunity for the encapsulated gyne to make the final cuts to the cell cap and emerge from its
cell. It may also be suggested that the swarming event itself may stimulate the emergence of encapsulated gynes.
For those colonies containing clipped queens, failed swarms ensued and in the cases of 2014 French colony 6
(Supplementary Fig. S66), 10 (Supplementary Fig. S67) and 12 (Fig. 11), the early tooting that occurs could also
be the consequence of a queen emerging due the swarming attempts, and not that of opening the hive, causing the
early release of an encapsulated gyne. It could therefore be considered that clipping queens causes havoc within
the natural swarming procedure of a colony. If made available to beekeepers, our swarming prediction alarm
would remove the necessity of queen clipping altogether, as well as minimise the need for other repeated invasive
procedures, and allow for colonies to develop more naturally.

Conclusions
To conclude, what we present here is a significant advancement in the detection of pre-swarming cues with the
use of averaged vibro-acoustic information to non-invasively monitor the swarming activities within honeybee
colonies, without the need for invasive visual observations. Through novel analytical techniques, we developed
two methods for the prediction of swarming in honeybee colonies. Each provides a generic signature that ena-
bles automatic assessments of a colony’s intent to swarm, further substantiating the usefulness of accelerometer
technology for the in-situ non-invasive monitoring of honeybee colony status. The timings of the toot and quack
signals that we have recorded, relative to those of swarms, strongly support the idea that tooting and quacking
enable the colony to regulate afterswarming and ensure that the parental colony is always left with at least one
mature gyne after a swarm has issued.

Methods
Continuous recording of vibrational data. Accelerometers were embedded in the wax at the centre of
the central frames of twenty-five honey bee hives containing various subspecies of Apis mellifera. All accelerom-
eters were placed among bee brood14. Twenty-two hives at the Institut National de la Recherche Agronomique
(INRA) were monitored using high sensitivity (100 mV/g) Brüel and Kjær Type 4508 piezoelectric accelerom-
eters. One colony at an apiary in Jarnioux, France and two at the Nottingham Trent University (NTU) Clifton
Campus, UK were monitored using ultra-high sensitivity (1000 mV/g) accelerometers of the same type. These
accelerometers are of exacting specification with a quoted frequency-dependent sensitivity variation of about 2%
over this range, an operational temperature range of −54 to 121 °C and a temperature coefficient of sensitivity of
+0.006% per 1 °C. The Brüel and Kjær software PULSE was used to process the signal digitised by the B&K Type
3053-B-120 conditioner, and collect these measurements continuously, in the form of power spectra averaged
every 3 minutes, with a bandwidth of 5500 Hz and a frequency resolution of 3 Hz. Over 30 million individual
frequency resolved data points were logged and analysed. The apiary at INRA was comprised of one Langstroth
and nineteen Dadant hives that were established two meters apart in a row approximately 40 meters in length in
Avignon, France, in March 2014. Additional accelerometers were also placed in the wax at the centre of seven
other frames within one of the NTU hives. All accelerometers were oriented so that their sensing direction was
perpendicular to the plane of the comb to obtain the greatest sensitivity. To install the accelerometers, a cavity (1
cm3) was made at the centre of the focal comb, an accelerometer was then placed inside, and a small amount of
molten wax was applied to aid in securing its position and prevent any direct exposure of the metal components
to the bees (Fig. 12).
Hives were continuously monitored for vibrations throughout the 2014 and 2015 seasons, apart from short
electrical power cuts after which the data loggers resumed automatically once power was restored. Each day,
the previous day’s spectrogram of vibration was generated to continuously monitor the progress of each of the
twenty-four hives throughout the entirety of their time under observation. An excellent array of swarming events
(a total of eleven primary swarms and nineteen afterswarms) were captured in the data across both the 2014 and
2015 seasons.

Swarming prediction using averaged vibrational spectra. Training Phase. Strategy 1 – Discrimination
between Instantaneous SpectraA specific swarming feature that worked over an entire season was not found
during the development of this algorithm. However, a specific signature was obtained when restricting the data
to the swarming season only.
Datasets from colonies that swarmed were selected through visual inspection of the daily spectrogram of
vibration (see Fig. 7 and Supplementary Fig. S74). To compile the average spectra to be used in the Preparing to

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Figure 12. A Brüel and Kjaer piezoelectric Type 4508 accelerometer installed in the centre of a frame from a
bee hive.

Swarm (PTS) training database, data for the thirteen days prior to the primary swarm were selected from each
hive that swarmed. For the Non-Swarming (NS) dataset, spectra gathered after the last queen pipe recorded in
the swarming colonies were further labelled as ‘not preparing to swarm’. Spectra were also compiled and added
to the training database from the entire swarming season (April – July) for those colonies that did not swarm. As
honeybees (Apis mellifera spp.) remain in their hives overnight, only data logged from midnight to 06:00 were
included to ensure they were not influenced by foraging activity, which upon inspection was found to yield highly
variable and unpredictable spectral information. Data were averaged to give a mean spectrum for each hour of
recording and were separately uploaded into the software. The data were then normalised and reshaped to create
a linear array which could later be fed into the PCA/DFA algorithm (for further information see e.g. Bisele, et al.28,
Bencsik, et al.14, Ramsey, et al.15,16) to identify spectral information unique to each category.
Strategy 2 – Discrimination between Spectral Evolutions.We also attempted a second, much more computation-
ally intensive, algorithm that included information about the spectral history in a procedure we term “spectral
evolution”. The basic concept behind this algorithm was to use the temporal evolution of the spectrum of vibra-
tion that takes place over the days leading up to a primary swarm, rather than relying on a single instantaneous
spectrum. To compute the spectral evolution, the FFT is calculated for the time course of the magnitude of all
the uploaded spectral frequencies for each day between midnight and 5:00 AM, yielding 2DFT images. The FFT
is then further calculated over each pixel of the series of 2DFTs found in the preceding days, the number of
which was explored during parametric optimisation, resulting in a 3D spectroscopy exercise. The power spectra
information in both the 2DFT and 3DFT revealed changes in the history of the spectra, without keeping the
absolute timings of the changes. This prevents the risk of training the algorithm to absolute timings. The 3DFTs
are then reshaped and compared to a series of discrimination curves identified through rigorous training of a
Discriminant Function Analysis (DFA) algorithm (see below) for categorisation of a colony as being either in a
swarming or non-swarming state.
To create these DF curves, two training databases were first established, made up of spectral information from
colonies that either did or did not prepare to swarm. The PTS database was comprised of data containing infor-
mation (on the ten-day spectral evolution) of the six days leading up to issuance of the primary swarm of eighteen
different datasets (11 colonies including one swarm made available from 8 independent accelerometer measure-
ments in separate frames) from the UK and French colonies in 2014 and 2015. The NS database was made up of
the 3D spectra from across the active season of fifteen colonies that did not swarm and the 3D spectra after the
final afterswarm of the ten colonies that did swarm. To compute the spectral evolution from a time period not
affected by the variation induced by foraging, the three-minute power spectra were uploaded each day between
the hours of midnight and 5:00 AM.
Principal Component and Discriminant Function Analysis.For both strategies, the training databases were
reshaped to create a linear array, normalised with respect to their mean and, only in the case of the instantaneous
spectra algorithm, log transformed to help scale-up minute differences in spectra. The databases then underwent
reduction using Principal Component Analysis (PCA) to create a series of PCA scores that best describe the data
deviations.
For each method, the categories were carefully selected within the training database and their PCA scores
were calculated. By using a triplet of cross correlation products with three discriminant functions identified by
the DFA algorithm (previously made available online, Bencsik, et al.15), three discriminant function coordinates,
or ‘DF scores’, could then be calculated for each measurement. The centroids for each group were then calculated.
From these, a “threshold value” for swarming detection could be determined for the ratio of distances from the
point in DF space to each centroid, considered the “alarm” value, between the PTS and NS centroids that dictated
whether a measurement was classified as being in a state of ‘preparing to swarm’ or ‘not preparing to swarm’. The
optimum threshold value was that which allowed the highest number of successful classifications. To reduce the
possibility of over-fitting, and to find the most generic discrimination functions, an iteration procedure similar to
that of Bisele, et al.28 was also implemented. Every pulse combination was explored within the training database,
removing all spectra except for a reduced collection that best represents the overall information pertaining to

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each category, allowing for clustering with the lowest overall misclassification. However, spectra removed from
the training database that were not included within the computer training stage were still included in the cluster-
ing process to find their DF space co-ordinates and contribute to the calculation of the overall classification error.
Parametric Optimisation.The numerical search that we have performed was entirely self-contained within the
MATLAB high level programming language, from data upload to the swarming signature extraction. The sig-
nal processing comprises numerous steps, all of which can be tailored by the user to refine the outcome of the
search. This allowed us to run optimisation loops, in which a specific parameter along the processing stages was
systematically changed, until the extracted swarming signature provided the best outcome, i.e. lowest overall
misclassification. This numerical exercise, which we call ‘parametric optimisation’, works given that we have a
scalar quantity that, at the end of a specific processing chain, provides an estimate of the performance or of our
swarming detection. This scalar quantity was determined by calculating the proportion of incorrectly classified
points; it defined what we call the percentage error.
To achieve optimisation, a number of parameters were explored to obtain the lowest percentage error of
discrimination between points pertaining to each group. Within the optimisation loop, parameters that were
changed independently from each other were: scaling (logarithm vs. linear spectral amplitude); the lower and
upper limits of the frequency bandwidth of the spectra contributing to the search; the number of days of data
leading up to the primary swarm included in the PTS training dataset; the specific hour over each day from
which the data were extracted; the number of PCA scores contributing to the DFA search (too low will remove
important information, too high will bring in ‘noise’ and overfitting artefacts); the specific spectra fed to the PCA
algorithm to get the PCA scores (this will dictate the relevance of the first PC scores, as discussed earlier); and
finally, the number of DF scores (two or three in our case) onto which the selected PCA scores are collapsed. To
dismiss the variation that sometimes arises due to the influence of the wind (see Supplementary Figs. S75 and
S76), frequencies below 18 Hz were systematically excluded. In the case of the strategy making use of the spectral
evolution, the number of days contributing to the spectral evolution was also explored.
We tested the algorithms on the swarming season datasets for all colonies. For each dataset, spectra were
averaged over each hour and then categorised as belonging either to a PTS or NS state, using a triplet of cross cor-
relation products with the two discriminant functions identified by the DFA algorithm. The threshold value acted
as the “alarm” which is triggered if a specific spectrum was deemed to be close enough to the swarming centroid.
The alarm value was plotted for each hour of the day throughout the swarming season. The exact time when a
primary swarm left the hive within each dataset was then also displayed. This allowed to visually assess how many
days prior to that time the alarm was triggered and to also identify any false positives or negatives.

Continuous in-hive monitoring of queen pipes. In 2015, an additional hive was equipped with a
1000 mv/g Brüel and Kjaer piezoelectric Type 4508 accelerometer for the exploration of raw accelerometer data.
The data pertaining to this hive, named the Jarnioux 2015 dataset, are fully described and partially available as
part of previous publications by Ramsey, et al.15,16. These data were intentionally transformed into daily spectro-
grams of individual spectra averaged over three minutes with parameters strictly identical to those used by the
PULSE software that stored the other datasets. In particular, the frequency resolution was set to 3 Hz whilst the
bandwidth was set to 5 kHz.
The spectral signature unique to each queen pipe was identified through critically listening to the raw audio
of the Jarnioux 2015 dataset (e.g. S1 Audio) and cross referencing this to the three-minute average spectra at
the associated time points. This allowed for visual discrimination between toots and quacks within vibrational
datasets comprising of three-minute averaged spectra. The occurrences of toots and quacks were then examined
visually within the daily images and presented alongside each other and other key colony-level events.

Received: 29 January 2020; Accepted: 15 May 2020;

Published: xx xx xxxx

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Acknowledgements
This work was undertaken as part of the EU funded Swarmonitor project. We gratefully acknowledge the
European Union’s Research Executive Agency for funding under project reference No 315146. We acknowledge
Claudia Dussaubat at INRA, David Parker, David Whittaker and Catherine Procter at NTU for their contribution
and to the Swarmonitor consortium members, the BFA, the EPBA, the CARI, CAPAZ, ARNIA and SZOMEL for
helpful discussions. We also thank NTU for funding the first author’s work in writing up this paper for publication.
We thank Joseph Bencsik for providing the bees and experimental set up for the Jarnioux measurements, and Bob
Smith for helpful discussions. We would finally like to thank Professor Gard Otis for the time and care taken to
review our paper, providing in-depth editorial suggestions.

Author contributions
Conceived and designed the experiments: M.B., Y.L.C., M.I.N. Performed the experiments: M.B., M.R., M.P., D.C.
M.I.N. analysed the data: M.T.R., M.B. Wrote the paper: M.T.R., M.B., Y.L.C., M.R., M.P., N.S.D., M.I.N. Colony
status analysis: M.R., D.C. All authors reviewed and edited the manuscript.

Competing interests
The authors declare no competing interests.

Additional information
Supplementary information is available for this paper at https://doi.org/10.1038/s41598-020-66115-5.
Correspondence and requests for materials should be addressed to M.B.
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