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Institution : Biologie & Ecologie Tropicale & Méditerranéenne de

Perpignan (UMR 5244), France

Premier cas documenté de transmission parasitaire dans le sud

de la France entre des espèces de tortues de Floride introduites

et les tortues d’eau douce indigènes (Emys orbicularis) et

Mauremys leprosa)

Olivier Verneau et Louis Du Preez

N2 N2 N2

N2

N1

N1

N1

AREA 1 AREA 2

E1

Rana catesbeiana Myocastor coypus

Instances of invasive species in France

Repartition of Emys orbicularis in Europe

Repartition of Emys orbicularis in Languedoc-Roussillon

Repartition of Emys orbicularis in Languedoc-Roussillon

Repartition of Mauremys leprosa in Europe

Repartition of Mauremys leprosa in Languedoc-Roussillon

•! Life Cycle of the Polystomatidae

Final host

Eggs

Larvae

Adults Bladder

Pharynx Palpebral

cavity

Polystomoides Polystomoidella Neopolystoma

Neopolystoma orbiculare (Stunkard, 1916) Price, 1939

Host = Trachemys scripta

Distribution = USA

Site = Bladder

Polystomoides megacotyle (Stunkard, 1916) Ozaki, 1935

Host = Trachemys scripta

Distribution = USA

Site = Pharynx

Neopolystoma euzeti Combes & Ktari, 1976

Host = Mauremys leprosa

Distribution = Tunisia

Site = Bladder

Polystomoides tunisiensis Gonzales & Mishra, 1977

Host = Mauremys leprosa

Distribution = Tunisia

Site = Pharynx

Polystomoides ocellatum (Rudolphi, 1819) Ozaki, 1935

Host = Emys orbicularis

Distribution = Europe occidentale and Morocco

Site = Pharynx

Paris

Perpignan

LEUCATE

LEUCATE

BANYULS

CANET

LEUCATE

BANYULS

St HIPPOLYTE

THUIR

CANET

LEUCATE

BANYULS

St HIPPOLYTE

THUIR

SOREDE

METHODOLOGIE

2. Noter le numéro de la tortue sur le bac

3. Isoler les tortues individuellement dans un peu d’eau. Les laisser au moins

une nuit.

4. Utiliser deux tamis, un de 500 microns (au dessus) et un second de 100

microns (au dessous). Verser l’eau du bac sur le tamis supérieur.

5. Rincer le tamis de 100 microns sur une boite de Petri

6. Verser le contenu de la boite de Petri dans un tube propre. Numéroter

le tube et l’envoyer à Olivier Verneau, UPVD

7. Répéter l’opération à partir de l’étape 3. au moins une fois si possible.

Verser le contenu de la boite de Petri dans le même tube.

8. Au labo, recherche des œufs du parasite sous une loupe binoculaire

‘Pear’ shaped egg

Bladder parasite

‘Diamond’ shaped egg

Palpebral parasite

•! Methodology - COI Sequencing

- Alignement

- NJ tree

•! Results - Polystome eggs (Emys and Mauremys)

- NJ tree (351 characters)

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

Bladder

Palpebral cavity

Pharyngeal cavity

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

1.15 % Pharyngeal

cavity

Polystomoides tunisiensis1

< 2% > 2%

One species Three species

Amphibian polystome species delineation

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

A

B

C

Bladder

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

New species

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

0.28 %

0.83 %

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

~7.0 % A

B

C

Bladder

0.28 %

0.83 %

Neopolystoma euzeti

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

0.7 %

Pharyngeal cavity

0.1

Polystomoides malayi

Polystomoides siebenrockiellae

Polystomoides coronatum

Polystomoides spEoL1

Polystomoides spEoL2

Polystomoides spEoL3

Polystomoides spEoL4

Polystomoides spEoL5

Polystomoidella sp

Polystomoides spEoS5

Polystomoides spEoS4

Neopolystoma spTs

Polystomoides spMlS3

Polystomoides spEoS1

Polystomoides spEoS2

Polystomoides spMlS1

Neopolystoma orbiculare1

Neopolystoma orbiculare2

Polystomoides spPn

Polystomoides spMlS2

Polystomoides oris1

Polystomoides oris2

Polystomoides spTs1

Polystomoides spTs2

Polystomoides megacotyle

Polystomoides spEoS3

Neopolystoma euzeti

Polystomoides asiaticus

Polystomoides tunisiensis3

Polystomoides tunisiensis1

Polystomoides tunisiensis2

Neopolystoma spratti

Neopolystoma fentoni

Neopolystoma spEoS1

Neopolystoma spCs

Neopolystoma spEoL6

Neopolystoma spGg

Neopolystoma spMlB1

Neopolystoma palpebrae

Neopolystoma liewi

Polystomoides bourgati

0 %

Palpebral cavity

Conclusions

•! En milieu confiné (Vallée des tortues)

Vessie Vessie

Chrysemys

Emys et Mauremys Trachemys

Emys

Conclusions

•! En milieu confiné (Vallée des tortues)

Pharynx Pharynx

Chrysemys Mauremys

Conclusions

•! En milieu naturel (Leucate)

- présence d’au moins une espèce

parasite indigène sur Emys (nouvelle

espèce de la vessie). 70% au moins des

tortues infestées

Cavités palpébrales Cavités palpébrales

Graptemys Emys

Conclusions

•! En milieu naturel (Banyuls, St Hippolyte)

- absence des espèces parasites

indigènes sur Mauremys

- présence d’au moins une espèce

parasite allochtone

Conclusions

•! En milieu naturel (Banyuls, St Hippolyte)

Cavités palpébrales Cavités palpébrales

Graptemys Mauremys

Emys (Leucate)

Conclusions générales

•! Risque sanitaire ?

•! Surveillance parasitologique

•! * Sampling effort per year (5, 6, 4 nights and 12 traps in 2006, 07 and 08 respectively

•! & Using test of Schnabel

CANET

BANYULS

St HIPPOLYTE

THUIR

Le BOULOU