Defending the nest against its archenemy: The Defending the nest against its archenemy: The Cyprian honeybee Cyprian honeybee Apis mellifera CypriaApis mellifera Cypria

confronting the Oriental hornetconfronting the Oriental hornetVespa orientalisVespa orientalis

PAPACHRISTOFOROU AlexandrosPAPACHRISTOFOROU Alexandros11,, RRORTAIS AgnORTAIS Agnèèss11, , SSUEURUEUR JJéérômerôme22, ARNOLD , ARNOLD GGéérardrard11

1 Laboratoire Evolution, G1 Laboratoire Evolution, Géénomes, Spnomes, Spééciation, CNRS Gifciation, CNRS Gif--sursur--Yvette, France, Yvette, France, 2 Mus2 Musééum National dum National d’’Histoire Naturelle, DHistoire Naturelle, Déépartement Systpartement Systéématique et matique et ÉÉvolution,volution,

Paris, FranceParis, France

Apis mellifera cypriaApis mellifera cypria

Annual Development of coloniesAnnual Development of coloniesApis melliferaApis mellifera

““CriticalCritical”” periodperiod:: Early SpringEarly Spring

Apis mellifera cypriaApis mellifera cypria

““CriticalCritical”” periodperiod: : Early AutumnEarly Autumn

AugustAugust--OctoberOctober�� Dry weatherDry weather--High High temperaturetemperature

�� Reduced populationReduced population--No broodNo brood

�� High density of hornetsHigh density of hornets’’attacksattacks

�� Limitation of flight Limitation of flight activityactivity ((Vespa Vespa orientalisorientalis--Merops Merops apiasterapiaster))

8 JULY

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Flight activityFlight activity

Vespa orientalis

Strong mandiblesStrong mandibles

Hard cuticleHard cuticle

Large sizeLarge size

Confronting hornets during Confronting hornets during attacksattacks

Movement of Movement of defending honeybeesdefending honeybees

METHODOLOGYMETHODOLOGY

““Arresting hornetsArresting hornets””

AnesthetizingAnesthetizing--Adjusting hornetsAdjusting hornetsMonitoringMonitoring

METHODOLGYMETHODOLGY

�� Image and video Image and video analysisanalysis ((ImageJ, ImageJ, Pinacle Liquid Pinacle Liquid Edition V6)Edition V6)

�� Seasonal effect of Seasonal effect of defensive defensive behaviourbehaviour

�� Graphs of moving Graphs of moving patterns in timepatterns in time((at flight boardsat flight boards))

Seasonal effect of defensive behaviourSeasonal effect of defensive behaviour

Defensive behaviour during hornetDefensive behaviour during hornet’’s attacks attack ((20 May20 May))

Movement of honeybees defending the Movement of honeybees defending the entrance of the colonyentrance of the colony

Numbers of honeybees participating defenseNumbers of honeybees participating defense

““Pattern APattern A””. Honeybees. Honeybees, , defending colonies defending colonies outside the hive. Moving towards intruderoutside the hive. Moving towards intruder

““Pattern BPattern B””. Honeybees. Honeybees, , retreating inside the retreating inside the hive. Avoiding contact with intruderhive. Avoiding contact with intruder

Propolis walls at the entrance of the colonyPropolis walls at the entrance of the colony

Propolis walls at the entrance of the colonyPropolis walls at the entrance of the colony

MethodologyMethodology

A) A) --Pictures of propolis wallsPictures of propolis walls--Estimation of each openingEstimation of each opening--Evaluation of hornetEvaluation of hornet’’s body section s body section (measured the size of the largest section (measured the size of the largest section of their thoraxof their thorax))--Evaluation of the Evaluation of the percentage of openings percentage of openings through which hornets can get inside the through which hornets can get inside the colonycolony

ΒΒ) ) Destroyed propolis wallsDestroyed propolis wallsRecording their reRecording their re--construction by construction by honeybeeshoneybees within a yearwithin a year

Percentage of openings through which Percentage of openings through which hornets can get inside the colonyhornets 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 ballingKilling the enemy during balling--SuffocationSuffocation

Current BiologyCurrent Biology 1717((1818): ): RR795795--796796

Killing the enemy during ballingKilling the enemy during balling

Killing the enemy during Killing the enemy during ballingballing--SuffocationSuffocation

�� ThermoThermo--ballingballing ((Ono Ono et alet al., 1995)., 1995)Vespa mandarinia Vespa mandarinia (47(47ooC)C)

Apis cerana Apis cerana (49(49ooC)C)

�� Vespa orientalisVespa orientalis –– Apis mellifera cypria Apis mellifera cypria ??

Killing the enemy during ballingKilling the enemy during balling�� Materials and methodsMaterials and methods::Monitoring: temperature inside the ballMonitoring: temperature inside the ball--Duration of ballingDuration of ballingHornets inside the incubatorHornets inside the incubator

�� ResultsResults::Temperature in balls:Temperature in balls: 4444οοCCTime required for a hornet to die:Time required for a hornet to die: 5757..88 minmin--Hornets inside incubator at Hornets inside incubator at 4444οοCCTime required for a hornet to die:Time required for a hornet to die: 143143..33 minmin--Upper Thermal Lethal LimitUpper Thermal Lethal Limit::Vespa orientalisVespa orientalis (50,(50,55ooC)C)Apis mellifera cypriaApis mellifera cypria (50,(50,66ooC)C)

Most honeybees attack the abdomen of the Most honeybees attack the abdomen of the model than the headmodel than the head--thoraxthorax

Models of hornetsModels of hornets--Monitoring hornetsMonitoring hornets’’ behaviourbehaviour

Spiracles ofSpiracles of Vespa Vespa orientalisorientalis

Monitoring the respiration rhythm of Monitoring the respiration rhythm of hornets in the labhornets in the lab

�� Adjustment of hornetsAdjustment of hornets�� Monitoring the normal respiration rhythmMonitoring the normal respiration rhythm

�� Monitoring the respiration rhythm during balling simulationMonitoring the respiration rhythm during balling simulation((Zafeiridou Zafeiridou & & TheophilidisTheophilidis 2006)2006)

Monitoring the respiration ofMonitoring the respiration of Vespa Vespa orientalis orientalis under normal conditions and under normal conditions and

during simulation of ballingduring simulation of balling

Insertion of plastic blocks under abdominal tergitesInsertion of plastic blocks under abdominal tergites

Assessment of balling duration while preventing hornets Assessment of balling duration while preventing hornets from blocking the abdominal function of hornetsfrom blocking the abdominal function of hornets

Time required for the death of hornetsTime required for the death of hornets

HeatHeat--balling or Asphyxiaballing or Asphyxia--balling?balling?�� Different speciesDifferent speciesApis ceranaApis cerana--Apis mellifera,Apis mellifera,Vespa mandariniaVespa mandarinia--Vespa orientalisVespa orientalis

Heat and carbon dioxide generated by honeybees Heat and carbon dioxide generated by honeybees jointly act to kill hornetsjointly act to kill hornets

““AbstractAbstract:: We have found that giant hornets We have found that giant hornets (Vespa mandarinia japonica) are killed in less (Vespa mandarinia japonica) are killed in less than 10than 10 min when they are trapped in a bee ball min when they are trapped in a bee ball created by the Japanese honeybees Apis cerana created by the Japanese honeybees Apis cerana japonica, but japonica, but their death cannot be solely their death cannot be solely accounted for by the elevated temperature in the accounted for by the elevated temperature in the bee ballbee ball……””

Sugahara MSugahara M & & Sakamoto FSakamoto FNaturwissenschaften 2009 96(9):1133Naturwissenschaften 2009 96(9):1133--11311366

Sounds emission during conflictSounds emission during conflict

ApidologieApidologie 3939::468468--474474

Materials and MethodsMaterials and Methods

�� Recording sounds atRecording sounds atWAV format (WAV format (Sennheiser Sennheiser MEME6464, , Marantz PMDMarantz PMD670670))--93 successful 93 successful recordingsrecordings

�� Recordings analysis with Recordings analysis with Seewave vSeewave v. 1.2 (. 1.2 (Sueur Sueur et et alal.., 2006), 2006)

�� Reproduction of soundsReproduction of sounds--Monitoring possible Monitoring possible effects on honeybeeseffects on honeybees’’behaviourbehaviour

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 frequencyDominant frequency (14(14 ff0): 60): 6..6060kHzkHz

Fundamental Fundamental frequency frequency ((ff0): 00): 0..4747kHzkHz

�� No differences No differences between between coloniescolonies

�� Absence of Absence of anacoustical anacoustical ““signaturesignature””

Comparison of acoustic parameters Comparison of acoustic parameters between 9between 9 coloniescolonies ((AA--II))

Reproduction of soundsReproduction of sounds--Monitoring Monitoring possible effects on honeybeespossible effects on honeybees’’ behaviourbehaviour

ColonyColony

Control 1Control 1

no soundno sound

2s natural 2s natural

sound 2s sound 2s

pausepause

ContinuantContinuant

Natural Natural

soundsound

Artificial Artificial

soundsound

66..6060kHzkHz

Control 2Control 2

no soundno sound

11 5353 3737 2424 2222 4848

22 9898 162162 136136 3030 4343

33 9696 7878 129129 3434 5757

44 412412 344344 454454 355355 575575

55 233233 243243 251251 126126 226226

66 6666 5252 3838 3333 5656

77 462462 389389 502502 403403 621621

88 105105 9494 133133 4040 7474

99 204204 216216 2222 117117 198198

1010 7878 9999 8383 2525 4646

AverageAverage 180180..77aa 171171..44aa 177177..22aa 118118..55aa 194194..44aa

ConclusionsConclusions

� 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

AcknowledgmentsAcknowledgments

�� Special thanks to:Special thanks to:

--Prof. George Theophilidis and Dr Georgia Prof. George Theophilidis and Dr Georgia

ZafeiridouZafeiridou

--Dr Max WatkinsDr Max Watkins

--The CYPRIOT BEEKEEPERS ASSOCIATIONThe CYPRIOT BEEKEEPERS ASSOCIATION