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Defending the nest against its archenemy: The

Cyprian honeybee Apis mellifera Cypria


confronting the Oriental hornet
Vespa orientalis

PAPACHRISTOFOROU Alexandros1, RORTAIS Agnès1,


SUEUR Jérôme2, ARNOLD Gérard1

1 Laboratoire Evolution, Génomes, Spéciation, CNRS Gif-sur-Yvette, France,


2 Muséum National d’Histoire Naturelle, Département Systématique et Évolution,
Paris, France
Apis mellifera cypria
Annual Development of colonies
ApisApis mellifera
mellifera cypria
“Critical” period: Early Autumn
“Critical” period: Early Spring
August-October
 Dry weather-High
temperature
 Reduced population-
No brood
 High density of hornets’
attacks
 Limitation of flight
activity (Vespa
orientalis-Merops
apiaster)
0
200
400
600
800
1000
1200
1400
1600
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2000
0:00
1:00
2:00
3:00
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5:00
6:00
7:00
8:00
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8 JULY

13:00
14:00
15:00
16:00
Flight activity

17:00
18:00
19:00
20:00
21:00
22:00
23:00
Vespa orientalis

Hard cuticle

Large size

Strong mandibles
Confronting hornets during
attacks
Movement of
defending honeybees
METHODOLOGY

“Arresting hornets”

Anesthetizing-Adjusting hornets
Monitoring
METHODOLGY
 Image and video
analysis (ImageJ,
Pinacle Liquid
Edition V6)
 Seasonal effect of
defensive
behaviour
 Graphs of moving
patterns in time
(at flight boards)
Seasonal effect of defensive behaviour

Defensive behaviour during hornet’s attack (20 May)


Movement of honeybees defending the
entrance of the colony

Numbers of honeybees participating defense


“Pattern A”. Honeybees, defending colonies
outside the hive. Moving towards intruder
“Pattern B”. Honeybees, retreating inside the
hive. Avoiding contact with intruder
Propolis walls at the entrance of the colony
Propolis walls at the entrance of the colony
Methodology
A) -Pictures of propolis walls
-Estimation of each opening
-Evaluation of hornet’s body section
(measured the size of the largest section
of their thorax)
-Evaluation of the percentage of openings
through which hornets can get inside the
colony
Β) Destroyed propolis walls
Recording their re-construction by
honeybees within a year
Percentage of openings through which
hornets can get inside the colony
December 2005

March 2006

May 2006

July 2006

August 2006
December 2005

March 2006

May 2006

July 2006

August 2006
Killing the enemy during balling-
Suffocation

Current Biology 17(18): R795-796


Killing the enemy during balling
Killing the enemy during
balling-Suffocation
 Thermo-balling (Ono et al., 1995)
Vespa mandarinia (47oC)
Apis cerana (49oC)

 Vespa orientalis – Apis mellifera cypria ?


Killing the enemy during balling
 Materials and methods:
Monitoring: temperature inside the ball-
Duration of balling
Hornets inside the incubator
 Results:
Temperature in balls: 44οC
Time required for a hornet to die: 57.8 min
-Hornets inside incubator at 44οC
Time required for a hornet to die: 143.3 min
-Upper Thermal Lethal Limit:
Vespa orientalis (50,5oC)
Apis mellifera cypria (50,6oC)
Models of hornets-Monitoring hornets’ behaviour

Most honeybees attack the abdomen of the


model than the head-thorax
Spiracles of Vespa
orientalis
Monitoring the respiration rhythm of
hornets in the lab

 Adjustment of hornets
 Monitoring the normal respiration rhythm
 Monitoring the respiration rhythm during balling simulation
(Zafeiridou & Theophilidis 2006)
Monitoring the respiration of Vespa
orientalis under normal conditions and
during simulation of balling
Assessment of balling duration while preventing hornets
from blocking the abdominal function of hornets

Insertion of plastic blocks under abdominal tergites


Time required for the death of hornets
Heat-balling or Asphyxia-balling?
 Different species
Apis cerana-Apis mellifera,
Vespa mandarinia-Vespa orientalis
Heat and carbon dioxide generated by honeybees
jointly act to kill hornets

“Abstract: We have found that giant hornets


(Vespa mandarinia japonica) are killed in less
than 10 min when they are trapped in a bee ball
created by the Japanese honeybees Apis cerana
japonica, but their death cannot be solely
accounted for by the elevated temperature in the
bee ball…”

Sugahara M & Sakamoto F


Naturwissenschaften 2009 96(9):1133-1136
Sounds emission during conflict

Apidologie 39:468-474
Materials and Methods
 Recording sounds at
WAV format (Sennheiser
ME64, Marantz PMD
670)-93 successful
recordings
 Recordings analysis with
Seewave v. 1.2 (Sueur et
al., 2006)
 Reproduction of sounds-
Monitoring possible
effects on honeybees’
behaviour
Two successive hisses produced during hornet attacks: spectrogram
(frequency vs. time vs. relative amplitude in relative dB)
and oscillogram (relative amplitude vs. time).
Comparison of hiss and buzz acoustic
characteristics.

Dominant frequency (14 f0): 6.60kHz

Fundamental frequency (f0): 0.47kHz


Comparison of acoustic parameters
between 9 colonies (A-I)

 No differences
between
colonies

 Absence of
anacoustical
“signature”
Reproduction of sounds-Monitoring
possible effects on honeybees’ behaviour
Control 1 2s natural Continuant Artificial Control 2
Colony no sound sound 2s Natural sound no sound
pause sound 6.60kHz
1 53 37 24 22 48
2 98 162 136 30 43
3 96 78 129 34 57
4 412 344 454 355 575
5 233 243 251 126 226
6 66 52 38 33 56
7 462 389 502 403 621
8 105 94 133 40 74
9 204 216 22 117 198
10 78 99 83 25 46
Average 180.7a 171.4a 177.2a 118.5a 194.4a
Conclusions
 The Cyprian honeybee Apis mellifera, demonstrates
an effective defensive behaviour against the
Oriental hornet Vespa orientalis
 The expression of its defensive behaviour has some
specificities compared to other species or
subspecies:
-Different and contrary defensive “patterns” within
the same apiary
-Effective use of propolis walls during defense
-No stinging
-Suffocating hornets to death in balls of workers
-Sound emission of high frequent sounds
Acknowledgments
 Special thanks to:

-Prof. George Theophilidis and Dr Georgia


Zafeiridou

-Dr Max Watkins

-The CYPRIOT BEEKEEPERS ASSOCIATION

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