Rabbit and man: genetic and historic approach

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Rabbit and man: genetic and historic approach

M Monnerot, Jd Vigne, C Biju-Duval, D Casane, Cécile Callou, C Hardy, F

Mougel, R Soriguer, N Dennebouy, Jc Mounolou

To cite this version:

Original

article

Rabbit and

man:

genetic

and historic

_approach

M

Monnerot

JD

_Vigne

C

_Biju-Duval

D

Casane

C Callou

C

_Hardy

F

_Mougel

R

_Soriguer

N

_Dennebouy

JC Mounolou

1

1 CNRS, Centre de _{Genetique M oléculaire,} F91198 Cif sur-Yvette Cedex; 2

CNRS, URA _1415, Laboratoire d’Anatomie _Comparee,

MNHN, F75005 Paris, France;

3

Estacion

Biologica de _Donana, Pabellon del Peru, Avenida _Maria-Luisa,

41043 Seville, Spain

Summary - New data on mitochondrial DNA _polymorphism in _Oryctolagus cuniculus confirm the existence of 2 maternal _lineages which are _{geographically} well _{separated. They}

provide evidence in favour of northern _Spain

_(and

_possibly southern

_France)

as a _refuge area for rabbit _{populations during} the last major glaciation. Osteological analysis leads

to the discrimination of _populations and the _recognition of discrete _{qualitative characters,} which provide additional markers to describe _{population diversity.} Characterization of different domains of mtDNA from ancient bones was used as a tool to resolve the _general question of the _origin of present populations. Results obtained from ancient and _present rabbits living in Zembra

_(Tunisia)

showed that the _{present-day population} has descended from animals _present on the island some _{2 000 years ago. Archaeozoological} data provide

evidence for their introduction _by Bronze Age, Punic or Roman _people. rabbit _/ mtDNA _/ osteology / ancient DNA _/ domestication

Résumé - Le lapin et l’homme : _{approche génétique} et _historique. L’étude _approfondie du _{polymorphisme} de l’ADN mitochondrial chez le _{lapin, Oryctolagus cuniculus, confirme} l’existence de _{2 lignées} maternelles bien _{séparées géographiquement:} au Sud de l’Espagne

pour l’une, dans le reste de l’Europe pour l’autre. Elle suggère que le Nord de l’Espagne, et éventuellement le Sud de la France, ont été une des zones refuge des populations de

lapins lors des dernières _{importantes glaciations.} Une _{analyse ostéométrique} a été menée

sur quelques populations. Elle a identifié des caractères discrets qui peuvent désormais

être _pris en _compte dans la _description de la diversité des _populations. La caractérisation

moléculaire de _{différentes régions} de l’ADN mitochondrial extrait à _partir d’os anciens a

permis d’appréhender la _question de l’origine, dans le temps, de populations de lapins site par site : les résultats obtenus à partir de matériel ancien et récent ont montré que les

de _préciser que l’introduction du lapin sur cette île a _pu se réaliser par les peuples de l’âge de _bronze, les _Puniques ou les Romains.

’

lapin / ADNmt _/ ostéologie / ADN ancien _/ domestication

INTRODUCTION

The

rabbit,

Oryctolagus

cuniculus,

has 2

_interesting

characteristics: on the one

hand,

wild

_populations

still exhibit

_ample

_genetic

_diversity,

on the

_other,

this

animal is one of the rare mammals

_originally

domesticated in Western

_Europe.

The means

by

which

_genetic

diversity

has been maintained remains to be

_explained

since,

in their _ecosystem, rabbits _may be a _prey for numerous

_predators

and are

affected

_by

_epizootic

diseases

_(such

as

_haemolytic

viruses or

_{myxomatosis).}

In their natural

_environment,

_populations

levels and

_equilibrium

are also

_dependent

on

human

_activities;

rabbits are often

_{destroyed following}

_major

_damage

to fields and

replanted

forests. Moreover these animals are also used for

_{hunting exploitation}

and

consequently

are often reintroduced in

_large

numbers in

_places

where _game reserves are created.

The

_hypothesis

for southern

_Spain

as the area of

_origin

for rabbits comes from the

discovery

of the oldest known rabbit fossil in Andalusia

_(6.5

million years,

Lopez-Martinez,

1989).

Later

_glacial

events

_(until

12 000 _years

_ago)

_{probably strongly}

affected the

_biogeography

of the rabbit in its

_original

distribution _area, ie Iberia and Mediterranean France.

_Recently

human interference has affected the

_dispersion

of this

_species

_through

_{transportation}

within this area and

_outside,

_eventually

accompanied by

domestication and

_restocking.

Previous studies on some

_polymorphic

markers

_(Ferrand

et

_al,

_1988;

Arana et

_al,

1989;

Biju-Duval

et

_{al, 1991;}

Van der Loo et

al,

1991)

have

_{already provided}

first-hand information about the extent and the distribution of

_diversity

in

_European

rabbit

_populations.

These

_preliminary

results on

populations

from

Spain

and France are in _agreement with the

_hypothetical

view described above since

_they

show that

genetic

diversity

is

_consistently

much

_larger

in

_populations

from southern

Spain

than in the other areas, but

they

do not _{prove it.} A more exhaustive

_analysis

of

_genetic

_diversity

_(including

different

_markers),

both in various

_populations

and in relation to the

_geographical

distribution of rabbits is _{necessary to support} the

proposed

hypothesis conclusively.

Moreover,

the

_knowledge

of the evolution of

populations,

through

time, at

_given

_sites, is essential to understand the

_origin

of the

_biogeography

of

_genetic

_diversity.

This can now be

_{appraised through}

an

_{archaeozoological approach complemented by}

a molecular

study.

Altogether,

present and

_{archeological}

data should

_help

to define the role man has

_played

in

rabbit dissemination.

New data on mitochondrial

polymorphism presented

here confirm the existence of 2 maternal

_{lineages, geographically}

well

_{separated. They}

_argue in favor of

north-ern

_Spain

_(and

southern

_France?)

as a

refuge

area for rabbit

populations during

discrete

_qualitative

characters which

_provide

additional markers to describe

popula-tion

_diversity.

Characterization of different domains of mtDNA from ancient bones was used as a tool to resolve the

_general

question

on the

origin

of present

popula-tions. Results obtained from ancient and _present rabbits

_living

in Zembra

_(Tunisia)

show that the

_{present-day population}

is the progeny of animals introduced on the

island at least 2 000 _{years ago.}

MATERIALS AND METHODS Animals

Wild

_Oryctolagus

cuniculus are from ’natural

populations’

ie

populations

without

evidence of recent

_import

of animals from elsewhere. Domestic rabbits were

ob-tained from INRA laboratories

_(New

_Zealand,

_California)

and the Institut Pasteur

(Burgundy,

Tunisian and

_Portugese

domestic

_types).

Table I indicates the

_origins

and numbers of all the animals

_studied;

those with an asterisk have

previously

been

described in

_Biju-Duval

et al

_(1991).

Techniques

DNA extraction

Mitochondrial DNA for restriction

_analysis

was extracted from soft tissues and

purified according

to

_Biju-Duval

et

_al,

1991.

Total DNA was extracted from frozen tissues

_following

Kocher et al

_(1989),

and from bones as

already

described

by Hardy

et al

_(1994).

Restriction

_analysis

and

_phylogenetic

trees

For each

_mtDNA,

the sites

_{recognized by}

14 restriction enzymes have been deter-mined and

_mapped.

In order to collate the

results,

the different mtDNA _types have been named

_according

to their maternal

_lineage

_(A

or B see further in the

_results)

and numbered

_following

their

_description.

This led us to rename the _types

already

presented by

Biju-Duval

et al

_(1991).

The

_{correspondence}

is as follows: Lo 1 to 7

will now be referred to as A 1 to

_7;

Se = B

8;

Tu =

B10;

Az =

B9;

Tv1 =

B4;

Tv2 =

_B5;

Ce =

_B3;

Fbl =

_B1;

Fb2 =

_B2;

Ze1 =

B6;

and Ze2 = B7.

Phylogenetic analysis

of restriction site

data,

based on

_parsimony,

was

_performed

using

the

_{phylogeny analysis}

_{using parsimony}

program

(PAUP,

Swofford,

1989).

Nucleotide distances were calculated

following

Nei and Li

(1979)

and standard errors estimated

through

the

jacknife

method

(Efron,

1979).

Dendrograms

were obtained

_through

the

_{neighbor-joining algorithm}

_(Saitou

and

Nei,

1987)

following

the program ’restsite’

(Miller, 1991)

Amplification

and

_sequencing

The conditions of

_{amplification}

within the 16s-rRNA gene,

sequencing

and

Amplification

within the

_cytb

gene was

_performed

_using

the

_primers

_cytb3

(L15367,

ATGAAACTGGCTCCAACAAC)

and thr3

_(H15915,

CTCCATTCCTG-GCTTACAAGAC)

and

_{accomplished by}

2 min at

_93°C,

followed

_by:

1 min at

_93°C;

1 min at

_55°C;

30 s at 70°C

_through

40

_cycles;

followed

_by

2 min at 72°C. The PCR conditions were: 10 mM Tris HCl

_pH

9.0 at

_25°C;

1.5 mM

_MgCl

_z

_;

50 mM

_KCI;

0.1% Triton

_X100;

150 mM

dNTPs;

and 1 mM

_primers.

An

_aliquot

was then submitted

to

_asymmetric

_{amplification}

with

_only

one

_primer

as follows: 2 min at

_93°C;

40 s at

_93°C;

33 s at

_55°C;

45 s at

_72°C;

35

_cycles.

The

_products

were

_{purified through}

phenol

chloroform extraction and ethanol

precipitation

(50%,

2.5 M ammonium

CCTTG).

Internal

_primers

_specific

for rabbit mtDNA were used for

_amplifying

and

_sequencing

ancient DNA:

_{cytb6, cytbll}

_(L15691,

ATGT

CTAAACAACGTAG-CATG),

cytbl2

(H15835,

CTTGCGAGGGGTATA

_AGAATA)

and

_cytbl3

_(H15765,

GCGACTTGTCCAATGGTGATG.

Dating

bones

The _ages of the

_layers

of the ancient bones of

_Oryctolagus

cuniculus were obtained

through Ly-4382

carbon

_dating

of the charcoals

_(Hardy

et

_al,

₁₉₉₄₎

and confirmed

by

the examination of

_potteries

in the

_{corresponding layers.}

Osteometric

_analysis

The main dimensions of skull and the

_lengths

of the limb _segments were

_appraised

using

12 distinct measurements and the discrete characters were

analyzed

as

previously

described

_(Vigne

et

_al,

in

_press).

RESULTS

Population analysis through

RFLP

_(restriction

_fragment

_length

poly-morphism)

Reexamination of the

_population

from Las Lomas

_(Andalusia,

southern

Spain)

A

_{preliminary sampling}

of the

_population

from Las Lomas had

_already

revealed noticeable

_polymorphism

_(Biju-Duval

et

_al,

_1991).

The aim of a new

study

of this

population

was to evaluate the

_significance

of the

_{sampling procedure}

and to more

accurately

describe the

_diversity.

The

_study

of 31 additional animals

_caught

alive

identifies 7 mtDNA _{types among} which

_only

one was new

_(A8).

We can conclude

that the

_diversity

_appraised

from the first

_sampling,

conducted 4 _years

_earlier,

reflects well that of the whole

_population.

Table II

gives

the distribution of mtDNA _types

_according

to the

_territory

of the

rabbits. The existence of several mtDNA _types

_(at

least 3 and sometimes

₄₎

in each

locality signifies

that a noticeable

_diversity

exists at the level of

_family

_grouping,

ie

the burrow.

Population

from

_Badajoz

_(Estramadur,

southern

_Spain)

Among

16 rabbit extracts

_analyzed,

5 mtDNA _types were

recognized.

Four of them

belong

to the A

_lineage

_{(see below):}

_A4,

_9, 10 and 11.

_A4,

the

_only

one

previously

described from Las

Lomas,

is

_represented

in this

_{sampling by}

one animal. This means that the

diversity

of this

population,

located more than 200 km north of Las

Lomas,

is also noticeable with little

_overlapping

with the mtDNA _types described there. Three animals exhibited the type B3 relevant to the other maternal

_lineage

Population

from Navarra

_(northern

_Spain)

Four sites were

_sampled

in

_Navarra,

less than 70 km _apart, with a total of 16 rabbits.

Due to the low number of animals from 3 of these sites

_(Arroniz,

_Castejon

and

Sesma)

it seemed more reasonable at _present to consider them all

_together

as

representatives

of one extended

_population.

Four mtDNA types were detected:

B8,

9,

10 and _11, all new, and

again

specific

to this area.

Populations

from France

Eight

individuals from

_Arjuzanx,

southwestern France

_(Landes)

and 2 from Donzere

(southeastern

France)

were studied. These last 2

_rabbits,

as well as 3 from

Arjuznax,

carried a mtDNA _type

_B1,

ie the _type found in domestic

_breeds,

whilst _type B3 was found in the other 6 rabbits.

Rabbit from Norwich

_{(United Kingdom)}

The mtDNA

_type

B1 was detected in the

_single

individual

_analyzed.

Phylogenetic relationships

inferred from RFLP

_analysis

The

_complete

set of data is

_given

in the

_appendix.

Each mtDNA _type is defined

by

the _presence or absence of each of the 110 sites

_recognized

and

_mapped.

Two

programs were used to establish

_phylogenetic

_{relationships}

between the 22 mtDNA

types

actually

described

_through

RFLP

_analysis:

Paup 30L,

which is based on

parsimony;

and

_{neighbor-joining,}

which is based on nucleotide

_distances.

In both

cases, results on mtDNA from

_Lepus

_europaeus and

_{Sylvilagus rufeseens}

were

included as out groups.

Figure

1a and b present the

_phylogeny

obtained which

confirms that

_{proposed previously}

with less data

_(Biju-Duval

et

_al,

_1991).

The rare differences are relevant to the shortness of some branches:

_{they clearly}

show 2 groups of mtDNA _types, referred to as A and B. The

divergence

between the

2 groups is

_{high enough}

to consider them as

_{representative}

of 2

_{well-separated}

Reexamination of mtDNA

_type

Bl

_{through sequencing}

As mentioned above and in a

_previous

work

_(Biju-Duval

et

_al,

₁₉₉₁₎

the mtDNA

type B1 was detected in animals of different

_origins:

various domestic breeds

(Burgundy,

New

_{Zealand, California,}

_Portugal

and

_Tunisia)

as well as natural

populations

(Versailles,

Cerisay,

Arjuzanx,

Donzère and

_Norwich).

This result was

surprising

in that these animals have various

_geographical

_{origins and,}

for the

domestic

_breeds,

are derived from _very different sets of crosses

(Arnold,

1981,

and Standard officiel des

_lapins

de race,

1984).

In order to

question

the apparent

homogeneity

of this group, we

_sequenced

the last _part of the

_cytochrome

b _gene. No variation was detected among the 522 nucleotides

sequenced

from mtDNA extracted

from different well-known domestic breeds:

California;

Burgundy;

New

_Zealand;

domestic animals from Tunisia and

_Portugal,

the race of which was not

_known;

and wild B1 animals

_(Cerisay,

Donzère and

_Arjuzanx,

see

_above).

Morphometry

Results

_given

_by

multivariate data

_analysis

_(ACP

and

_AFD)

of Las Lomas

popu-lations have shown that factors of total

_length

and breadth of the _{cranium may} well characterize a rabbit

_population

(Vigne

et

al,

in

_press).

_Although

such an

analysis

on other

populations

(Zembra,

Domestic,

Camargue)

is not _yet

complete,

a

preliminary approach

has validated them for

population comparison.

For

example,

individuals of southern

_Spain

_(Las

_Lomas)

can be

assigned

as small type whereas

rabbits of the

_Camargue

are

_larger

than average.

Eight

original qualitative

discrete characters were found

_(Vigne

et

_al,

in

_press)

most of them

_being

related to the

_posterior

_part of skull: the

_shapes

of’ the

’scutellum’

_(DS1);

the crista nuchae

_{(DS2, DS3);}

and the

_foramen

magnum

(DS4).

According

to the

_recognition

of their presence or

absence,

the animals from Las

Lomas,

Zembra and domestic breeds can be

_{distinguished}

(table III).

For

_example,

DS4a is

_{significantly}

absent in domestic rabbits. The combination of some characters

may also be informative

(table IV).

DSlb-DS2a and DSlb-DS2b are the

only

ones

observed in domestic

_rabbits,

the first

_being

in the

_majority,

whereas rabbits from Zembra exhibit many others.

Furthermore,

the

frequency

of each discrete character within the

_population

can also be taken into account. The use of discrete characters

Ancient DNA

Archaeozoological

surveys on Zembra island

_(Tunisia)

_provided

30 fossiliferous sites.

The

_oldest,

which dates from the

_Upper

Pleistocene when the island was linked to the

_mainland,

testifies to the absence of rabbit in the north-eastern

_{Maghreb during}

this

_period.

Sediments dated from the end of the Neolithic or at the

_beginning

of the Bronze

_Age,

when Zembra was

already

_isolated,

also did not

_provide

any rabbit remains. The oldest bones come from late Roman

_layers

dated from the 3rd-4th centuries

_AD,

and other more recent sites have confirmed the _presence of

rabbits on the island

_during

the later

_periods

_(Vigne,

_1988;

_Hardy

et

al,

in

_press).

Unfortunately,

no Punic or

_early

Roman

_desposits

have been found _yet. Present data

indicate that rabbit was introduced to Zembra between the Late Neolithic and the 3rd _century AD

_(ie

_by

Bronze

_Age,

Punic or Roman

_people).

A

_{preliminary analysis}

of mtDNA

_(part

of the 16S-rRNA

_gene)

from ancient bones

_(dated

back to 130-390

_AD)

has demonstrated that the

_{corresponding}

animals carried _type B mtDNA

(Hardy

et

_al,

_1994).

A more detailed

_study

has been conducted in order to more

precisely

show which B _type was _present in these animals.

_Preliminary

_sequencing

of different domains of the

_cytochrome

b _gene have shown that the last _part was one of the most variable ones

_(Howell,

_1989, and present

work)

and revealed a

clear distinction between mtDNAs of _types B1 and B7. Two

_overlapping

_fragments

were

_{amplified, using}

_cytb6-cytbl3

and

_{cytbll-cytbl2,}

and

_{sequenced. Among}

the

190bp examined,

4

_{non-contiguous}

_nucleotides,

located at sites

_926,1016,

1023 and

1038

_(numbered

from the

_beginning

of the

_gene)

_appeared

variable within _type B

mtDNAs.

_They

are all different when B1 mtDNA

_(from

domestic

_stock)

is

_compared

to B7

_(from

_Zembra):

_T,

_{A, A,}

G instead of

C,

G, G, A, respectively.

mtDNA

amplified

from ancient bones

_(the

same as in the

_previous

_study,

see

above)

exhibits

the _pattern

_{C, G, G, A,}

which is identical to the mtDNA from rabbits

_presently

living

on Zembra.

_{Consequently,}

we believe that these animals are descended from

those present on the island almost 2 000 _{years ago.}

DISCUSSION

Origin

of

_European

rabbit

_populations

As shown in

_figure

_2,

the _present

_diversity

of mtDNA is

_organized

as follows.

whereas all the animals from northern

_Spain,

_France,

Norwich and Zembra

_(Tunisia)

carry mtDNAs of

lineage

B.

_{Paleontologic}

data indicate the

_origin

of

_Oryctolagus

genus in southern

Spain

6-6.5 million years ago

(Lopez-Martinez,

1989).

The value

of the

_divergence

between molecules from

_lineage

A and

_lineage

B

_(4.5%)

and the

_geographical

_organization

of

_diversity

_{(fig 1)}

_support

the

_proposal

that the diversification within each

_lineage

was established

independently

about 2 million years ago

(based

on a

divergence

rate of

_2%/million

years, see discussion in

Biju-Duval et

_al,

_1991).

This means that after the

_{original dispersion}

of animals over

Spain,

the ancestors of

_{populations actually}

_carrying

either _type A or

_type

B molecules must have been

_separated

_{(geographically)}

some time _{before the age}

of the ancestor molecule of each

_{lineage. Following}

this

_scheme,

_progeny of one

sub-group

remained in southern

_Spain

_(A)

when the other was first restricted to northern

_Spain

_(B,

where the maximum of

_diversity

is

_found).

More

_recently,

the latter

_spread

over Western

Europe

was

_possibly

achieved

_by

human interference

(see

below).

Data on nuclear

variability

are in _agreement with this

description.

The

number of alleles at the b-locus of the

_{immunoglobulin light}

chain constant

_region

is far

_higher

in southern

_Spain

than in the

_population

from

_Camargue,

while rabbits

from northern

_Spain

exhibit an intermediate situation

_(Van

der Loo et

_al,

_1991;

Van der

_Loo,

_personal

_{communication).}

_Although

there is some

_{discontinuity}

in

rabbit

_samplings

in central

_Spain

and

_Portugal,

the

_recognition

of northern

_Spain

as a

_region

with

_specific

_properties

is sustained

_by

a recent work of Palomo et al

(in

press).

Taking

into account

_geographic,

climatic and

_geological

_parameters, as well as _present

_species

_diversity,

these authors demonstrate that 2 domains can be

defined in the Iberian

_peninsula:

a northern _part

_{corresponding}

to about a _quarter of

the

_peninsula,

and a central-meridional

_region

roughly

south of the 42nd

parallel.

Thus,

the

_recognition

of northern

_Spain

as a

_{plausible refuge}

area for maternal

lineage

B in rabbits can now be

_integrated

in a more

general understanding

of

species

history.

The

_origin

of the 3 rabbits from

_Badajoz

with _type B mtDNA deserves

_special

attention. An

_hypothetical

_import

from northern

_Spain

or France

_by

man seems

unlikely.

The

_high

_density

of rabbits in southern

_Spain

and

_{Portugal precludes}

_any need for massive

_{reintroduction,}

but restricted introduction cannot be excluded. At

present, another

_{plausible explanation}

is that Los

_Quintos

is _part of the

_secondary

contact zone

_along

which rabbits from maternal

_lineages

A and B meet

_again

after

some time of isolation.

Recent

_history,

the role of man

From the late Pleistocene until Classic

_Antiquity,

rabbits

_{only occupied}

the Iberian

peninsula

and a narrow area in southern France

(Donnard, 1982).

As a _consequence

of

_{transportation}

_by

_man, the

species

is now

_represented

in a

_large

_part of

_Europe,

in

South

_America,

in Australia and _many oceanic islands. The colonization of

_Europe

is the

_only

one

_preceeding

the 17th _century

(Angerman, 1974).

_{Archaeozoological}

data on the colonization _process are

incomplete

and must sometimes be treated with caution.

_Dating

_may suffer some uncertainties due to the

_burrowing

habits of

rabbits.

_However,

3

major

phases

can be

_recognized.

Throughout

antiquity,

the

_Phenician,

Greek and Roman

_people

may have

played

the introduction of rabbits in various

_regions

of the western Mediterranean Basin

by

the establishment of

Leporaria,

ancestors of our Middle

Age

warrens

(Bodson,

1978;

Rougeot,

1981). Archaeozoological

data are sometimes in _agreement with this view but not

_always.

For

_example,

in

_Zembra,

the absence of bones in a

Late Neolithic

layer,

the _age of the oldest rabbit

_bones,

3rd - 4th centuries AD

(Vigne,

1988;

Hardy

et

_al,

₁₉₉₄₎

_together

with the evidence that Punic

_people

occupied

the island from the 6th to the 2nd centuries BC

_(Chelbi

and

Ghalia,

personal

communication)

indicate that the introduction of the

_species

must have

been

_{accomplished by}

Bronze age, Punic or Roman

_{people. According}

to ancient

mtDNA studies

_(Hardy

et

_al,

1994 and _present

_report)

it can even be said that

rabbits

_presently

found on the island are the _progeny of those

_living

on the island at least 1 600 - 1 900 _{yr ago.} On the other

hand,

in

Corsica,

the presence of rabbits

in historical times was inferred

_(Bodson,

_1978;

_Rougeot,

_1981;

_Arthur,

₁₉₈₉₎

from the name ’cunicula7iae’

_given

_{by Pliny}

the Old

_(Nat

hist

_3,

₈₃₎

to the islands off

Bonifacio

_and/or

from

_Polybius’

_writings

_(Nat

hist

_{12, 3)}

In

_fact,

recent excavations

have shown the absence of rabbits in

_Tyrrhenean

islands at that time

_{(Vigne, 1992).}

From

_this,

it can be assumed that ancient authors confused

Oryctolagus

cuniculus

with another

_Lagomorph

_species:

_Prolagus

_sardus,

_presently

_extinct, but abundant

in Corsica and Sardinia at that time.

Most of the diffusion of rabbit in southern France and other western

_European

from the

_early

Middle

_Ages

which

_probably

have to be related to the

_production

of

‘Laurices’

_{(Rougeot, 1981),}

texts indicate that the

expansion

was

_only

_important

in

the 9th - 12th centuries which is confirmed

_{by osteoarchaeological}

data

_(Rougeot,

1981;

Delort, 1984;

Audoin,

1986).

Around that time

_{(10th century)}

in

_France,

medieval warrens,

previously

restricted to

_nobles,

were

developed throughout

the country

(Gislain,

1980; Zadora-Rio,

1986).

They

remained _open until mid-13th

century,

allowing

some individuals _{to go} back to the fields and found new

_colonies,

completely independent

of human interference and are

_probably

the

_origin

of _present warren rabbits. Animals were also

propagated through

sales from one _country to another and from

_gifts

to

_foreign

lords

_(Delort,

1984; Durliat,

1985).

Domestication

began

with the 16th _century

_(Rougeot,

_1981;

_Audoin,

₁₉₈₆₎

while warrens were

conserved

_(Zadora-Rio,

_1986).

After this

_period,

3 kinds of rabbits were

living

in Western

_Europe:

wild rabbits

_(most

of them

_coming

from

_warrens),

rabbits

_kept

in warrens for

_hunting,

and domestic stocks

_(Audoin,

_1986).

Thus the most

_important

diffusion

_through

Western

_Europe

took

_place

between

the 11th and 16th centuries

_AD,

while the

_species

was not domesticated but

appropriated

for

_hunting

_(Vigne,

in

_press).

Man

_managed

to

_keep

rabbit as a wild animal for the

_symbolic

value of rabbit

_hunting

_(Houseman,

_1990;

_Poplin,

in

press)

and as a

_prestige

and _power instrument of a

_privileged

social class

(ie

noble

people).

It seems clear that the absence of domestication of this

species

during

the

antiquity

led to the dissemination of rabbits as wild animals. This human behavior is

_responsible

for the foundation of wild

_populations,

which can still be found all over Western

_Europe,

because

keeping

them in a strict domestic status would have

consequently

limited their dissemination.

Origin

of the rabbits taken for domestication

The

_analysis

of restriction sites of mtDNA from a few races has revealed a

_surprising

homogeneity: they

all _carry the B1 _type. This _type has also been detected in the rabbit from Norwich and in the

_population

from

_Versailles,

both

_{being examples}

of

animals

_{transported by}

man at different times

_(the

16th century for Versailles and

probably

the llth - 12th centuries for

_Norwich)

and also in animals from natural

populations

(Cerisay,

Arjuzanx

and

_Donzère).

A more detailed examination

_by

DNA

_sequencing

confirms this result and no variation was detected in the 8 individuals

_analyzed

in a variable

region

of the

cytochrome

b gene. Such an absence of

variability

_may be

interpreted

in 2 _ways. One _may

_imagine

that domestic

_breeds,

some of them

being

more than 200 _yr old

(Arnold,

1981),

as well as rabbits

transported by

man from the llth to the 16th century were all issued from the same location. This is

_unlikely

since it is known

that some domestic stocks were established

independently

in different

_regions

and even in different countries.

Another,

more

plausible, explanation

is that man has

sampled

individuals at different times and

_places

from

_populations

with a _very low

polymorphism.

This would mean that

by

the llth _century, at the

_beginnning

of the

propagation

of rabbits in non-Mediterranean _areas, animals

_carrying

mtDNA _type

B1 were the most

_frequent.

A third

_possibility

is that animals

_carrying

maternal

ACKNOWLEDGMENTS

Thanks are due to H De _{Rochambeau, INRA, Castanet-Tolosan,} for _providing domestic animals. We also wish to thank the Office National de la Chasse for the access to the natural _populations it _supervises, 0 Ceballos and D Bell for their _help in _{getting samples}

from Navarra and _{Norwich, respectively.} This work has been _{financially supported by} Paris-Sud _University and _by a _{specific grant} from Bureau des Ressources G6n6tiques.

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