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Reintroduction plan for the endangered Sicilian black bee Apis mellifera siciliana

Cecilia Costa1, Carlo Amodeo2, Marco Lodesani1, Stefano Reale3, Raffaele Dall’Olio1

1 Consiglio per la Ricerca e la sperimentazione in Agricoltura – Unità di ricerca di Apicoltura e bachicoltura, via di Saliceto 80 – 40128, Bologna, Italia2 Apicoltura Amodeo di Amodeo Carlo, C.da Madonna Diana 90018 Termini Imerese (PA), Italia3 Istituto Zooprofilattico Sperimentale della Sicilia, Via G. Marinuzzi 3 –90129, Palermo, Italia

Apis mellifera geographic variability

� 26 described honey bee subspecies� Some distributed worldwide, some risk extinction

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Italian honey bee subspecies

A.m.ligustica A.m.siciliana

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A. m. siciliana

� Genetic characteristics from the African branch A and/or the Oriental branch O

� Characteristic behaviour and development

� Suffered from change in beekeeping after the 1960s (agricultural market and policies and varroa)

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Italian honey bee subspecies

A.m.siciliana A.m.ligustica

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• Mid 80s: «pure» coloniesassessed by morphometry and allozyme (MDH & EST) analyses

• Establishment of conservationisland: Ustica

• 1990s: conservation with risk of inbreeding

• 2000-2001: new conservationislands Filicudi & Vulcano; National Breeders Registry

• Separation of different lineson different island

• Within subspecies diversity by maintaining and crossingdifferent lines

The conservation programme of A. m. siciliana

Carlo Amodeo

FILICUDI

USTICAVULCANO

Munoz et al, Effectiveness of conservationislands, in review

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The Slow Food international organization:

• founded by Carlo Petrini in 1986

• Slow Food believes that everyone has a fundamental right to the pleasure of good food and consequently the responsibility to protect the heritage of biodiversity, culture and knowledge that make this pleasure possible.

• GOOD, CLEAN and FAIR

• involves more 100,000 people from 153 countries of five continents

The Slow Food foundation for Biodiversity

• promotes preservation of agriculturalbiodiversity through taste education

• promotes and coordinates over 300 ‘Presidia’ (200 of which are Italian) in the world

Sicilian Black Bee (2009)

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3 years - 400K€funding from SicilianRegional Government

Dec 2011 –Dec 2014

SOAT CollesanoApicoltura AmodeoIZS Palermo

Consiglio per la Ricerca e sperimentazione in apicoltura

– Unità di ricerca di apicoltura e bachicoltura

"APESLOW: Reintroduction and conservation of the endangered subspecies Apis mellifera siciliana (DallaTorre, 1896): a tribute to Prof. P. Genduso."

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How to achieve this ?

� Large scale reproduction of geneticmaterial (distribution of queen cells)

� Extensive sampling and analyses to findnew sources of genetic variability

� Identify areas suitable for controlledmating

� Team building and sharing knowledgeand experience

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In the first year and a half of the project ….

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• Constitution of the new beekeeperassociation for Sicilian black beebreeders

• Training course for queen breeding

• Rearing and distribution of queen cells

• Landscape study (beehiveslocation/new mating stations)

• Sample collection and analyses and sharing of techniques

31 founding members, currently 78

Beekeepers join association and apply for queen cells

Assignment criteria (location, VET registration)� selected beekeepers

Cells production and distribution � mating in different areas

Distribution of queen cells

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Reintroduction: spreading A.m.siciliana genes

2012 2013

N° beekeepers 49 78

N° queen cells 3207 6287

N° breeders 6 7

N° breeding lines 16 16

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Searching for more A.m.siciliana

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Recognition of A.m.siciliana

A79 ♣ ♦

A7 ♣

A043 ♦

A29 ♦

A88 ♣ ♦

Ap43 ♣

A8 ♣ ♦

Ap55 ♣

A14 ♦

A113 ♣ ♦

A(B)124 ♣

Ap226 ♦

Microsatellite DNA - 12 loci

A.m.siciliana vs Mediterranean races � DBase Univ.Murcia (♣)

(De la Rùa P., Dall’Olio - 2009)

A.m.siciliana vs European races � DBase Univ.Aarhus (♦)

(Kryger P. - 2010)

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274 samples x 5 bees = 1370 bees

Microsatellite analyses: results

Each sample: individual assignmentsof the 5 bees

� GENEALEX: frequency of correctassignment

� STRUCTURE: conservative cut-off level based on mis-assigned in reference DB frequency of correct assignments& mean % assignment to differentpopulation

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0%

5%

10%

15%

20%

25%

30%

35%

40%

45%

Recognition of A.m.siciliana

•Conservation islands (41)•100% A4

•Other samples (52)• 6 different haplotypes

• 100% “A”

• A9 rare, known in A.m.intermissa

A4 (75)

A3 (9)

A1 (4)A10 (2)

A11 (2)

A9 (1)

mtDNA – 1 bee/colony PCR-RFLP (DraI)

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Mitochondrial DNA

Recognition of A.m.siciliana

� 84 samples, 10-14 wings

� 16 wing characters (11 angles, 4 dimensions, 1 ratio)

� CRA-API reference database (176 samples, 6 subspecies)

� Post hoc discriminant analyses 0%

5%

10%

15%

20%

25%

30%

35%

40%

45%

Probability class

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Morphometric analyses

Selection criteria for conservation / reintroduction

Microsat mtDNA Morphometry Rare haplotype / private alleles

Apiary use ≥ suff > 70%

Mating station ≥ good A > 90%

Conservationisland

≥ excellent A > 90% yes

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1) PALERMO CITTÀ2) CASTELBUONO 3) RISERVA DELLO STRASATTO

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1

2

Establishing mating stations

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Increasing conservation islands

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Salina-Lipari

Marettimo

Linosa

Special acknowledgement to:

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Founders of Associationand breeders:

� Sergio Sapienza

� Fortunato Battaglia

� Giulio Vitale

Ongoing support:

� Eugenia Oliveri

NO COLLABORATION = NO CONSERVATION

Thank you for your attention !

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cecilia.costa@entecra.itraffaele.dallolio@entecra.it

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