CESAM - FCUL

Portuguese

beekeeping reality and the new

challenges in Varroa control

A. S. Lima, M. Vilas-Boas, A. C. Figueiredo

Talvipäivät29th January 2017, Hämeenlinna

2

Portugal

Azores

Madeira

- Population: 10 millions

- Area: 92 000 Km2

- 5th largest E.E.Z. in EU (1 727 408 km2)

- Mediterranean - Atlantic climate

average 13-18ºC winter/ 28-35ºC summer

- Sea surface 13-23ºC

- Hours of sunshine: 4-6h W/ 10-12h S

- Very rich biodiversity (fauna and flora)

3

Beekeeping in Portugal

- National Federation of Portuguese Beekeeping, since 1996

40 local beekeeping associations

16 cooperatives and honey producers organizations

pay annual fee of 125€

2017-2019 budget: 2 598 517€

- Beekeepers need to:

do registration on the Portuguese National Authority for Animal Health

register, and update every year, the nº of colonies in apiary

follow some rules to establish the apiary

4

Beekeeping in Portugal

Legislation DL 203/2005:

- one apiary can’t have more than 100 hives in one place

- must be establish 50m far from the road and 100m far from houses

- if more than one apiary in the same place:

100 m

up to 10 hives

5

Beekeeping in Portugal

Legislation DL 203/2005:

- one apiary can’t have more than 100 hives in one place

- must be establish 50m far from the road and 100m far from houses

- if more than one apiary in the same place:

400 m

from 11 up to 30 hives

6

Beekeeping in Portugal

Legislation DL 203/2005:

- one apiary can’t have more than 100 hives in one place

- must be establish 50m far from the road and 100m far from houses

- if more than one apiary in the same place:

800 m

from 31 up to 100 hives

7

Beekeeping in Portugal

Balance

2007 2010 2013 2016 (nº and %)

Nº of beekeepers 15 267 17 291 16 774 10 698 -4 569 -30%

Nº of apiaries 32 685 38 203 40 176 33 876 1 191 4%

Nº of colonies 555 049 562 557 566 793 626 339 71 290 13%

Total of apiaries/ beekeeper 2,1 2,2 2,4 3,2

Total of colonies/ beekeeper 36 33 34 59 In: FNAP Reports

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Beekeeping in Portugal

8% beekeepers

Average 9 apiaries/ beekeeper

139 colonies/ beekeeper

34% beekeepers

9

Beekeeping in Portugal

1 063

Beekeepers Apiaries Colonies

367 608

13 602

In: FNAP Report 2015

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Beekeeping in Portugal

Hobbyist beekeepers

Apiaries

Colonies

Beekeepers

In: FNAP Report 2015

11

5%

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Beekeeping in Portugal

- Apis mellifera iberiensis

- Mixture of clonal variation of African (A) and west European (M) lineages

- Same body size as European subspecies

- Very vigorous and active

- Supports long and cold winters

- Great collectors of propolis

- High propensity to swarm

- Very defensive stings a lot

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Beekeeping in Portugal

- 9 Protected Denomination of Origin for honey (EC 509/2006)

- honey is the main product of Portuguese beekeeping

in 2007: ± 7 000 ton in 2015: ± 11 500 ton (average 16-22 Kg/ hive)

export 2 000 ton (17% of national production)

import 1 321 ton

average price wholesale

3,30 €/Kg (multifloral)

4,10 €/Kg (monofloral)

5,50 €/Kg (organic)

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Beekeeping in Portugal

Hive products

Foundations10-20€/Kg

70-100€/Kg8-12€/Kg

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Beekeeping in Portugal

Controlled zones

- established since 2005 (DL 203/2005)

- systematic disease control is done

- prior authorization of all type of beekeeping material to be used

- constant vigilance and epidemiological prevention

- 2016 18 controlled zones (32% of mainland is under surveillance)

43%

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Beekeeping in Portugal

Mandatory to declare:

Tracheal mite

A. woodis

VirusDWV

Varroa mite

V. destructor

American foulbrood

P. larvae

NosemaN. cerana

N. apis

European foulbroodM. plutonius

chalkbroodA. apis

Tropilaelaps mites

SHBA. tumidaAFB EFB Nosema Tracheal mite Varroa Chalkbrood

0

5

10

15

20

25

30

35

40

2011201220132014

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Beekeeping in Portugal

Emerging pest (invasive)

A. mellifera

Vespa crabro

Vespa velutina nigrithorax

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Beekeeping in Portugal

- Varroa destructor:

- 1st time detected in 1986

- is the greatest threat to Portuguese beekeeping

- sanitary treatments of the colonies represents at least 35% of costs

- Since 1997, there is a National Beekeeping Program promoting varroa control

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Fluvalinate

ThymolAmitraz

Formic acid

ThymolMenthol1,8-cineoleCamphor

± 30% Varroa resistance

Varroa authorized treatments

Flumethrin

Oxalic acid

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Plants as alternative

- Phytochemicals as an alternative and natural resource of acaricidal compounds

- comproved biological activity of plants

- Several advantages: biodegradable, low cost harvest, short environmental persistence, minimum risk for pest resistance development

- Essential oils, - plant extracts, - deccoction waters, - hydrolates…

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Essential oils

A.S. Lima (2013)

CBV/FCUL (2007)

- secondary metabolites are produced by specialised plant structures (trichomes)

- highly complex mixtures of volatile compounds (terpenoids)

- different modes of action (neurotoxic, respiratory chain)

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≈4000 taxa

385 endemic150

157

78 33141006

1233

A.S. Lima (2013)

CBV/FCUL (2007)

Portuguese flora

Impact and challenges

- Negative impact in Apiculture worldwide

- CCD- Hive environment is shared by honeybees and Varroa

- Acaricides are costly, ineffective and harmful for the ecosystem

- Understand the mechanism of Varroa host finding

- Find a new, sustainable and effective way of mite control

- Understand the mechanism of Varroa host finding (1)

- Find a new, sustainable and effective way of mite control (2)

Main goals of the work

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Research Institutions

25

Research Institutions

550Km

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Isolation of the EOs

A.S. Lima (2013)

EOs isolated by hidrodistillation using a Clevenger-type

apparatus

Chemical analysis performed by GC e GC-MS

27

Sampling beesFr

om th

e fie

ld…

…to

the

lab

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Bioassays with EOs

Complete exposure method:- acetone as solvent- EOs solutions (v/v): 0.25%, 0.5%,

1%, 2.5%, 5%- glass Petri dishes (100 x 15mm)- 5 emergent or adult bees naturally

infested/ Petri dish- 5 replicates/ EO concentration- Blank, positive and negative controls- 30±1ºC, 70% RH

- results after 6, 12, 24 and 48h

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Bioassays with EOs

- Emergent bees are more sensible than the adult ones

- Overall, the 2,5%concentration test,was the better one

toxicity bees vs acaricidal activity

EO time effect

- Low Varroa mortality on the blank and negative controls ( 3% and 7%), suggesting that EO are responsible for the real mortality

- Statistics analysis (model Probit, SPSS): LC50 e LC90

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EO4899 EO4899

EO4891 EO4891

EO5068 EO5068

Bioassays

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Bioassays

EO5714

EO5663

EO5714

EO5663

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EO5763

EO5749

EO4309

EO4444

EO5763

EO5749

EO4309

EO4444

Varroa sensing chemical cues

pupae larvae food

forager bees

nurse bees

5th instar larvae

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Sensory organs of Varroa

A. Wild (2010)

http

://w

ww

.ala

.org

.au/

wp-

cont

ent/

uplo

ads/

2011

/03/

varr

oa.jp

g

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Dillier, F.X. et al. (2006)

Sensory organs of Varroa

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Electrotarsography (EAG)

Prof. Doctor Victoria Soroker

Institute of Plant Protection

The Volcani Center

(Israel)

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10x

Electrotarsography (EAG)

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Time (5 s)

Resp

onse

am

p.

(0,2

mV)

-0,421 -0,439 -0,506

Pulse flow 1000 mL/min

Adapted from Eliash, N. et al. (2014)

at least 10 replicates measurements

Electrotarsography (EAG)EOs dose-response (v/v):0.1% 0.5% 2.5%0.25% 1% 5%

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Electrotarsography (EAG)

Lima, A.S. et al., Understanding the chemosensing response of Varroa destructor to volatile compounds of Portuguese aromatic plants. (in preparation)

acet 0,1 0,25 0,5 1 2,5 5 acet0

20

40

60

80

100

120

EO4309

Concentration(%) (v/v)

Resp

onse

am

plitu

de (%

)

acet 0,1 0,25 0,5 1 2,5 5 acet0

20406080

100120140

EO4899

Concentration (%) (v/v)

Resp

onse

am

plitu

de (%

)

acet 0.1 0,25 0,5 1 2,5 5 acet0

20

40

60

80

100

120

140

EO5068

Concentration (%) (v/v)

Resp

onse

am

plitu

de (%

)

acet 0,1 0,25 0,5 1 2,5 5 acet0

20406080

100120140160

EO5714

Concentration (%) (v/v)

Resp

onse

am

plitu

de (%

)

39

the EOs activated the mite chemosensory sensilla eliciting a positive-dose dependent effect in Varroa chemoreceptors.

Electrotarsography (EAG)

Lima, A.S. et al., Understanding the chemosensing response of Varroa destructor to volatile compounds of Portuguese aromatic plants. (in preparation)

acet 0,1 0,25 0,5 1 2,5 5 acet0

20

40

60

80

100

120

140

EO4444

Concentration (%) (v/v)

Resp

onse

am

plitu

de (%

)

acet 0,1 0,25 0,5 1 2,5 5 acet0

20406080

100120140160

EO5749

Concentration (%) (v/v)

Resp

onse

am

plitu

de (%

)

40

Interestingly two other EOs elicited negative‑dose dependent response.

Behaviour bioassays

100 mm

HB + EOHB Varroa

- 6 conc. EOs- 3 replicates (30

Varroas)- 33ºC, 70% RH- Obs. 1h, 2h e 3h

Lima, A.S. et al., Understanding the chemosensing response of Varroa destructor to volatile compounds of Portuguese aromatic plants. (in preparation)

41

Lima, A.S. et al., Understanding the chemosensing response of Varroa destructor to volatile compounds of Portuguese aromatic plants. (in preparation)

Behaviour bioassays

Correlation between Varroa behaviour and EAG dose –response effect of

EO’s

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Lima, A.S. et al., Understanding the chemosensing response of Varroa destructor to volatile compounds of Portuguese aromatic plants. (in preparation)

Behaviour bioassays

Other mechanisms of action may be evolved in

Varroa behaviour

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Have we already found a solution for Varroa control?

- Acaricidal formulations with promising EOs- Trap devices (seeking for possible collaboration with some Veterinary Pharmaceutical Companies)

- Upgrade scale: field tests

Not yet…

…but we are still working and we are getting closer!

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Acknowledgements

A. S. Lima is grateful to Fundação para a Ciência e Tecnologia (FCT) for PhD grant SFRH/BD/76091/2011. This study was partially funded by FCT, under PTDC/CVT-EPI/2473/2012, Pest OE/AGR/UI0690/2011 and UID/AMB/50017/2013.

Prof. Doctor Ana Cristina Figueiredo

CESAM – FCUL

Prof. Doctor Miguel Vilas-Boas

Prof. Doctor Victoria Soroker

MSc. Nurit EliashDoctor Nitin Singh

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Thank you for your

attention.

Kiitos huomiostasi.