Diversity of introduced terrestrial flatworms in the Iberian Peninsula: A cautionary tale

Abstract

Many tropical terrestrial planarians (Platyhelminthes, Geoplanidae) have been introduced around the globe. One of these species is known to cause significant decline in earthworm populations, resulting in a reduction of ecological functions that earthworms provide. Flatworms, additionally, are a potential risk to other species that have the same dietary needs. Hence, the planarian invasion might cause significant economic losses in agriculture and damage to the ecosystem. In the Iberian Peninsula only Bipalium kewense Moseley, 1878 had been cited till 2007. From that year on, four more species have been cited, and several reports of the presence of these animals in particular gardens have been received. In the present study we have: (1) analyzed the animals sent by non-specialists and also the presence of terrestrial planarians in plant nurseries and garden centers; (2) identified their species through morphological and phylogenetic molecular analyses, including representatives of their areas of origin; (3) revised their dietary sources and (4) used Species Distribution Modeling (SDM) for one species to evaluate the risk of its introduction to natural areas. The results have shown the presence of at least ten species of alien terrestrial planarians, from all its phylogenetic range. International plant trade is the source of these animals, and many garden centers are acting as reservoirs. Also, landscape restoration to reintroduce autochthonous plants has facilitated their introduction close to natural forests and agricultural fields. In conclusion, there is a need to take measures on plant trade and to have special care in the treatment of restored habitats.

Full text

Submitted 27 April 2014
Accepted 26 May 2014
Published 10 June 2014
Corresponding author
Eduardo Mateos, emateos@ub.edu
Academic editor
Hannah Buckley
Additional Information and
Declarations can be found on
page 30
DOI 10.7717/peerj.430
Copyright
2014
´
Alvarez-Presas et al.
Distributed under
Creative Commons CC-BY 4.0
OPEN ACCESS
Diversity of introduced terrestrial
flatworms in the Iberian Peninsula:
a cautionary tale
Marta
´
Alvarez-Presas
1
, Eduardo Mateos
2
,
`
Angels Tud
´
o
1
, Hugh Jones
3
and Marta Riutort
1
1
Departament de Gen
`
etica, Facultat de Biologia i Institut de Recerca de la Biodiversitat (IRBio),
Universitat de Barcelona, Barcelona, Spain
2
Departament de Biologia Animal, Facultat de Biologia, Universitat de Barcelona, Barcelona,
Spain
3
Department of Zoology, The Natural History Museum, London, UK
ABSTRACT
Many tropical terrestrial planarians (Platyhelminthes, Geoplanidae) have been intro-
duced around the globe. One of these species is known to cause significant decline
in earthworm populations, resulting in a reduction of ecological functions that
earthworms provide. Flatworms, additionally, are a potential risk to other species that
have the same dietary needs. Hence, the planarian invasion might cause significant
economic losses in agriculture and damage to the ecosystem. In the Iberian Peninsula
only Bipalium kewense Moseley, 1878 had been cited till 2007. From that year on, four
more species have been cited, and several reports of the presence of these animals in
particular gardens have been received. In the present study we have: (1) analyzed the
animals sent by non-specialists and also the presence of terrestrial planarians in plant
nurseries and garden centers; (2) identified their species through morphological and
phylogenetic molecular analyses, including representatives of their areas of origin;
(3) revised their dietary sources and (4) used Species Distribution Modeling (SDM)
for one species to evaluate the risk of its introduction to natural areas. The results
have shown the presence of at least ten species of alien terrestrial planarians, from all
its phylogenetic range. International plant trade is the source of these animals, and
many garden centers are acting as reservoirs. Also, landscape restoration to reintro-
duce autochthonous plants has facilitated their introduction close to natural forests
and agricultural fields. In conclusion, there is a need to take measures on plant trade
and to have special care in the treatment of restored habitats.
Subjects Biodiversity, Ecology, Genetics, Zoology
Keywords Platyhelminthes, Tricladida, Alien species, Habitat restoration, Soil fauna,
Molecular identification
INTRODUCTION
Most animal invasive species detected in Europe are terrestrial invertebrates (Roques et
al., 2009). Invading edaphic organisms can have dramatic effects on the environment,
due to the direct effects on native soil organisms, and through their interactions with the
environment aboveground. However, overall, their impact in human health and economy
How to cite this article
´
Alvarez-Presas et al. (2014), Diversity of introduced terrestrial flatworms in the Iberian Peninsula: a cautionary
tale. PeerJ 2:e430; DOI 10.7717/peerj.430

Figure 1 Distribution map of the terrestrial flatworms. (A) Subfamily Bipaliinae. (B) Subfamily Geo-
planinae. Information from http://turbellaria.umaine.edu.
is greater than their ecological impact (Vil
`
a et al., 2010). Among these organisms, land
planarians are becoming an important and diversified group of introduced species in
Europe.
Terrestrial planarians (Platyhelminthes, Geoplanidae) are divided into four subfamilies
(Bipaliinae, Microplaninae, Geoplaninae and Rhynchodeminae) with a cosmopolitan
distribution (Winsor, Johns & Yeates, 1998); however, most species are found in the
southern hemisphere. Bipaliinae (Fig. 1A) is absent from the American and European
continents, Geoplaninae (Fig. 1B) have an exclusively Central and South American
distribution, while Microplaninae (Fig. 2A) and Rhynchodeminae (Fig. 2B) are the
subfamilies with the most northerly distribution, including Europe. Terrestrial planarians
are the only free-living Platyhelminthes that do not live in an aquatic habitat. However,
they have not developed the capacity to prevent water loss and are thus strongly dependent
on environmental moisture levels (Froehlich, 1956; McDonald & Jones, 2007). They seem
to withstand this limitation through behavioral strategies such as hiding in damp refuges
during the day and becoming active during the night. Due to these characteristics, these
animals are considered to have a low capacity to disperse. In fact, in their areas of origin,
although a few species are well-adapted to open and human-transformed lands (Baptista &
Leal-Zanchet, 2010), most species are restricted to humid forest areas.
A total of 36 species of terrestrial planarians are known to have been introduced in
different countries around the globe. Most of these species have a big effect on terrestrial
´
Alvarez-Presas et al. (2014), PeerJ, DOI 10.7717/peerj.430 2/35

Figure 2 Distribution map of the terrestrial flatworms. (A) Subfamily Microplaninae. (B) Subfamily
Rhynchodeminae. Information from http://turbellaria.umaine.edu.
ecosystem processes because they prey on soil invertebrates (see references in Winsor,
Johns & Barker, 2004). So far, five of these species are considered to be either invasive and
cause problems with local biodiversity (Platydemus manokwari De Beauchamp, 1963),
or horticultural pests (Arthurdendyus triangulatus (Dendy, 1894)) or earthworm farm
pests (Bipalium adventitium Hyman, 1943; Bipalium kewense Moseley, 1878; Dolichoplana
striata Moseley, 1877, see Winsor, Johns & Barker, 2004).
In Europe there is evidence of the presence of at least 18 introduced terrestrial planarians
(Minelli, 1977; Ball & Reynoldson, 1981; Jones, 1988; Jones, 1998; Mateos, Giribet &
Carranza, 1998; Faubel, 2004; Jones et al., 2008; Vila-Farr
´
e et al., 2008; Vila-Farr
´
e et al.,
2011; Justine et al., 2014). In the Iberian Peninsula (IP) there are only a few published
records of introduced terrestrial planarians, and the only species cited are Bipalium
kewense in Barcelona (Filella-Subir
`
a, 1983), Platydemus sp. in M
´
alaga (Vila-Farr
´
e et
al., 2011), Obama sp. in Asturias (Fern
´
andez et al., 2013) and Rhynchodemus R02 and
Caenoplana coerulea Moseley, 1877 in Girona (Mateos et al., 2013). The last species has also
been cited in Menorca (Breugelmans et al., 2012).
After receiving multiple reports from non-scientists on the presence of “large and
colored” terrestrial flatworms in several localities in the IP, and given their observed
locations, particularly in private gardens, we decided to analyze their presence in garden
centers and plant nurseries.
´
Alvarez-Presas et al. (2014), PeerJ, DOI 10.7717/peerj.430 3/35

The aims of this work were to: (1) estimate the number of terrestrial flatworm species
introduced in the IP, and find their region of origin; (2) check whether plant nurseries
and garden centers are acting as entrance gates and reservoirs; (3) estimate the invasive
potential of some introduced species by considering their diet and by using Species
Distribution Modeling (SDM); (4) propose measures to prevent their becoming invasive
and to prevent further introductions and spread.
MATERIAL AND METHODS
Specimen collection
Specimens were sampled from four sources (Tables 1 and 2): (1) gardens, (2) nurseries and
plantations, (3) semi natural areas, and (4) from other countries (either the original area
of distribution or other invaded areas). Specimens from sources 1 and 2 were either sent
by people who knew our work through the information in social networks, or sampled
by us (all the localities reported by non-scientist collaborators correspond to gardens).
Specimens from source 3 were sampled by us. Specimens from source 4 were sent by
colleagues, specialists of the group, to whom we requested material for comparison with
the Iberian populations.
Data from a total of 13 domestic gardens, seven nurseries, two plantations (all confined,
humanized locations), and three semi natural areas (humanized environments that are
not confined and in direct contact with agricultural and forest areas) have been analyzed
(Table 1). The three “semi natural areas”, located in North-eastern Iberian Peninsula, were:
(1) Cal Tet, Parc Natural Delta del Llobregat, Barcelona (Fig. 3, Loc-code O); (2) Can
Cabanyes, Granollers, Barcelona (Fig. 3, Loc-code M); (3) Viaducte de Rubi
`
o, Vall d’en
Bas, Girona (Fig. 3, Loc-code P). In all three places recent habitat restoration activities
have been performed, including the transplantation of autochthonous plant species from
commercial nurseries.
Amateur collaborators photographed the animals alive and fixed them in absolute
ethanol. Specimens we collected were also photographed and external morphological
characters recorded. Subsequently, animals were subjected to two different procedures to
proceed to the species identification: (1) specimens for molecular analyses were fixed in
100% ethanol and (2) specimens for histological studies were killed with boiling water,
fixed with 10% formalin for 24 h, and then preserved in 70% ethanol.
Morphological studies
Preserved specimens were examined under a stereo microscope and notes of their
dimensions, appearance, color (though this is affected by preservation), eyes, any stripes
or pattern, the position of the pharyngeal aperture (mouth) and gonopore, if present,
were taken. Specimens with no visible gonopore were considered to be immature. It
was possible to identify some specimens, even immature ones, to species level without
further examination. For unrecognized specimens, or where identity was uncertain and
required confirmation, a mature specimen (evidenced by an open gonopore) was selected
and divided into various portions, being embedded in wax. The copulatory apparatus
´
Alvarez-Presas et al. (2014), PeerJ, DOI 10.7717/peerj.430 4/35

Table 1 Localities where introduced species have been found/recorded in the Iberian Peninsula. Data organized chronologically. Sampling code:
(fs), specimens from field surveys conducted by us in gardens, nurseries and semi natural areas; (sbp), specimens sent by people who knew our work
through the information in social networks; (bd), bibliographic data. Date in format yyyy/mm/dd. Collectors: AG, Alberto Gayoso; AL,
´
Alvaro Leal;
AT,
`
Angels Tud
´
o; CC, Cristina Cabrera; CI, C
´
esar de In
´
es; CS, Carmen Soler; EM, Eduardo Mateos; GG, Georgina Gratac
´
os; IV, Iv
´
an Salvia; JM,
Jacobo Mart
´
ın; MR, Marta Riutort; RS, Roberto S
´
aez; VS, Vicent Sancho; Montilivi-WEB, http//www.iesmontilivi.net/WebProfes/jbarbara/web/
Galeria/Imatges/Invertebrats/cuc.htm; XB, Xavier B
´
ejar.
Sampling
code
Loc
code
Locality Position Habitat Species Date Collector/Ref
bd A Caldes d’Estrac
(Barcelona)
N41.569467
E2.526316
garden Bipalium kewense 1983 Filella-Subir
`
a, 1983
fs B Barcelona
(Barcelona)
N41.398539
E2.142162
garden Bipalium kewense 1995 MR
sbp C Lourizan
(Pontevedra)
N42.410111
W8.667716
nursery Bipalium kewense 1990 AG
bd D Girona
(Girona)
N41.964541
E2.827842
garden Bipalium kewense 1994 Montilivi-WEB
sbp E Villamalea
(Albacete)
N39.362159
W1.601281
nursery Bipalium kewense 1998 VS
sbp F B
´
etera
(Val
`
encia)
N39.604153
W0.507864
garden Bipalium kewense 1999 VS
bd G Benarmargosa
(M
´
alaga)
N36.8248
W4.1809
mango
plantation
Rhynchodemini Ri1
G
2007/12/25 Vila-Farr
´
e et al., 2011
as Platydemus sp
sbp H Badalona
(Barcelona)
N41.460177
E2.243985
garden Caenoplana Ca1
G
2008 RS
bd I Menorca
(Balearic Islands)
N39.95000
E3.850000
orchard Caenoplana coerulea 2009/04 Breugelmans et al.,
2012
fs J Torruella de Fluvi
`
a
(Girona)
N42.17559
E3.03953
garden Obama sp.6
G
2010/04/04 MR
sbp K
`
Oliva
(Valencia)
N38.910550
W0.073200
garden Caenoplana Ca1
G
2010/11/08 VS
sbp L Ames
(A Coru
˜
na)
N42.857955
W8.653278
garden Caenoplana Ca1 2010/12/10 AG
fs M Granollers
(Barcelona)
N41.570240
E2.270532
semi natural Caenoplana Ca1
G
Kontikia ventrolineata
G
2011/02/28
2012/10/12
CS
EM
sbp N Boadilla del Campo
(Madrid)
N40.405270
W3.877014
garden Caenoplana Ca1 2011/10/15 JM
fs O El Prat de Llobregat
(Barcelona)
N41.309519
E2.120887
semi natural Caenoplana Ca1
G,M
2011/11/05 EM & CC
fs P Vall d’en Bas
(Girona)
N42.125939
E2.433678
semi natural Caenoplana Ca1
G
Rhynchodemus Rs1
G,M
2011/11/12
2011/11/26
EM & XB
EM & MR
fs Q Gav
`
a-1
(Barcelona)
N41.288100
E2.006233
nursery Obama sp 2012/03/13 AT & MR
fs R Gav
`
a-2
(Barcelona)
N41.293222
E2.017583
nursery Obama sp
G
2012/03/14 AT & MR
fs S Vilassar de Mar
(Barcelona)
N41.497084
E2.376178
nursery Obama sp
G
2012/03/28 AT & MR
fs T Tortosa
(Tarragona)
N40.767329
E0.556963
nursery Obama sp
G
2012/04/04 AT
(continued on next page)
´
Alvarez-Presas et al. (2014), PeerJ, DOI 10.7717/peerj.430 5/35

Table 1 (continued)
Sampling
code
Loc
code
Locality Position Habitat Species Date Collector/Ref
sbp U Treto
(Cantabria)
N43.392385
W3.470387
garden Obama sp
G
Bipalium kewense
G
2012/06/27 CI
fs V Bordils
(Girona)
N42.034804
E2.898615
nursery Caenoplana Ca1
G
Caenoplana Ca2
G,M
Caenoplana bicolor
G
Obama sp
G
Dolychoplana striata
G
Bipalium kewense
2012/10/22 EM
sbp W Girona
(Griona)
N42.009800
E2.825554
garden Caenoplana coerulea 2013/09/11 GG
sbp X Polop
(Alicante)
N38.622149
W0.126626
garden Caenoplana coerulea 2014/02/01 AL
sbp Y C
´
artama
(M
´
alaga)
N36.748333
W4.586944
garden Obama sp 2014/03/01 IV
Notes.
G
Species with genetic sequences.
M
Species sectioned for internal anatomy study (see Table 2).
(gonopore) and a small anterior region were sagittally and transversely sectioned at 10 or
15 µm, respectively, stained in Harris’ haematoxylin and eosin and mounted in Canada
balsam.
DNA extraction, gene amplification and sequencing
A small piece of tissue fixed in absolute alcohol was digested with Wizard Genomic DNA
Purification lysis Buffer (Promega, Madison, WI, USA) and Proteinase K overnight at
37
◦
C, following manufacturer’s instructions. The rest of the tissue is kept as voucher in the
Genetics Department (Universitat de Barcelona).
We amplified an approximately 1 kb fragment of the mitochondrial cytochrome c
oxidase I (Cox1 gene) and a fragment of approximately 1,500 bp of the 28S rRNA gene
(28S) by PCR reaction. PCRs were carried out in a volume reaction mixture of 25 µl. For
Cox1 we used primers BarS (
´
Alvarez-Presas et al., 2011) and COIR (L
´
azaro et al., 2009)
and conditions were as in
´
Alvarez-Presas et al. (2011); 28S rDNA gene was amplified in
two different overlapping fragments using the primers 28S1F, 28S4R, 28S2F and 28S6R,
and conditions as in
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort (2008). Amplification products
were purified with a vacuum manifold (Multiscreen HTS Vacuum Manifold; Millipore
Corporation, Billerica, MA, USA). DNA sequences were determined from both strands
using Big-Dye Terminator (3.1, Applied Biosystems, Foster City, CA, USA) and the
reaction products were separated on the ABI Prism 3730 automated sequencer (Unitat
de Gen
`
omica dels Centres Cient
´
ıfics i Tecnol
`
ogics de la UB).
PCR products of the 28S gene for some individuals, that yielded double bands in
the direct sequences, were cloned using HTP TOPO TA Cloning Kit for Sequencing
(Invitrogen) in order to be sure that only one type of sequence was recovered (since the
existence of a duplication of the ribosomal cluster is known in terrestrial planarians,
Carranza et al., 1996). The sequences of the clones showed that these bands corresponded
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Alvarez-Presas et al. (2014), PeerJ, DOI 10.7717/peerj.430 6/35

Table 2 Sequenced specimens. To each new sequence a three digit numeric code was assigned. Sequences from the GenBank database do not have specimen code
numbers, only when there are more specimens from the same species in the same locality was a specimen code assigned (three letters + one number). Loc codes are as
described in Table 1. Collector: DB, Dani Boix; EM, Eduardo Mateos; HJ, Hugh Jones; KA, Miquel Arnedo; LL, Laia Leria; LW, L Winsor; MA, Marta
´
Alvarez-Presas; SG,
S Graham; MV, Miquel Vila.
Species/morphotype Code Locality/ref. or Loc code or collector and position GenBank Code
28S COI
Family Geoplanidae
Subfamily Bipaliinae
Bipalium sp. Japan/Bagu
˜
n
`
a et al., 2001–
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort, 2008 DQ665959
a
AF178307
a,c
B. adventitium Leignston (USA)/Bagu
˜
n
`
a et al., 2001–
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort,
2008
DQ665956
a
AF178306
a,c
B. kewense 894 Ponta Delgada, Illa S
˜
ao Miguel (Ac¸ores, Portugal)/
DB N37.745196 W25.667408
*
KJ599731
a
KJ659612
*,a,c
621 Treto (Cantabria, Spain)/U
*
KJ659703
*,a
KJ659609
*,a,c
623 KJ659610
*,a,c
666 Bordils (Girona, Spain)/V
*
KJ659611
*,a,c
B. multilineatum South Korea /GenBank Direct submission2010-may-18 HM346600
c
B. nobile HM346598
c
Novibipalium venosum Japan/
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort, 2008 DQ666048
c
Subfamily Microplaninae
Microplana nana Vall d’en Bas (Girona, Spain)/Mateos et al., 2009 KJ599722
a
FJ969947
a,c
M. groga Canyamars (Barcelona, Spain)/Mateos et al., 2009 KJ599721
a
FJ969964
a,c
M. terrestris Pontedeume (A Coru
˜
na, Spain)/Mateos et al., 2009 KJ599724
a
FJ969952
a,c
Subfamily Rhynchodeminae
Tribe Caenoplanini
Arthurdendyus triangulatus Wainui Barrys Bay (NewZealand)/
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort, 2008 DQ665953
a
DQ666027
a,b,d
Artioposthia sp. Australia/Bagu
˜
n
`
a et al. 2001–
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort, 2008 DQ665954
a
AF178325
a,b,d
A. testacea Malborough (Australia)/Bagu
˜
n
`
a et al. 2001–
´
Alvarez-Presas, Bagu
˜
n
`
a &
Riutort, 2008
DQ665952
a
AF178305
a,b
Australoplana sp. New Plymouth, Omata (NewZealand)/
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort,
2008
DQ665955
a
DQ666028
a,b,d
Caenoplana sp. 1 -/
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort, 2008 DQ665964
a
DQ666031
a,b,d
Caenoplana sp. 4 -/
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort, 2008 DQ666032
a,b
Caenoplana bicolor 654 Bordils (Girona, Spain)/V
*
KJ659705
*,a
KJ659648
*,a,b
Caenoplana coerulea s.l. New Plymouth, Omata (NewZealand)/
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort,
2008
DQ665961
a
DQ666030
a,b
Menorca (Balearic islands, Spain)/(Breugelmans et al., 2012) JQ514564
a,b
(continued on next page)
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Table 2 (continued)
Species/morphotype Code Locality/ref. or Loc code or collector and position GenBank Code
28S COI
Tal1 Tallaganda (Australia)/Sunnucks et al., 2006 DQ227621
b
Tal2 DQ227625
b
Tal3 DQ227627
b
Tal4 DQ227629
b
Tal5 DQ227631
b
Tal6 DQ227632
b
Tal7 DQ227633
a,b
Tal8 DQ227635
a,b
Vic1 Victoria (Australia)/Sunnucks et al., 2006 DQ465372
a,b
Caenoplana morph Ca1 399 El Prat de Llobregat (Barcelona, Spain)/O
*
KJ659613
*,a,b
400 KJ659614
*,a,b
402 KJ659615
*,b
403 KJ659616
*,b
404 KJ659617
*,b
415 Vall de’n Bas (Girona, Spain)/P
*
KJ659618
*,a,b
416 KJ659619
*,b
417 KJ659620
*,b
418 KJ659621
*,b
419 KJ659622
*,b
420 KJ659623
*,b
421 KJ659624
*,b
422 KJ659625
*,b
423 KJ659626
*,b
424 KJ659627
*,b
443 Badalona (Barcelona, Spain)/H
*
KJ659700
*,a
KJ659633
*,a,b
444 KJ659634
*,a,b
445 KJ659635
*,b
446 KJ659636
*,b
450 KJ659637
*,b
451 KJ659638
*,b
452 KJ659639
*,b
453 KJ659640
*,b
454
`
Oliva (Val
`
encia, Spain)/K
*
KJ659641
*,a,b
(continued on next page)
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Table 2 (continued)
Species/morphotype Code Locality/ref. or Loc code or collector and position GenBank Code
28S COI
601 Garden, Adelaide (Australia)/SG S34.988611 E138.599722
*
KJ659702
*,a
KJ659642
*,a,b
603 Garden in Townsville (Palmetum, Australia)/
LW S19.260277 E146.822222
*
KJ659643
*,a,b
605 KJ659644
*,a,b
634 Nursery in Liverpool (UK)/HJ N53.3525 W2.902777
*
KJ659645
*,a,b
649 Granollers (Barcelona, Spain)/M
*
KJ659646
*,a,b
650 KJ659647
*,a,b
735 El Prat de Llobregat (Barcelona, Spain)/O
*
KJ659651
*,a,b
Caenoplana morph Ca2 426 Bordils (Girona, Spain)/V
*
KJ659628
*,b
427 KJ659629
*,b
428 KJ659630
*,b
430 KJ659699
*,a
KJ659631
*,a,b
431 KJ659632
*,a,b
657 KJ659649
*,a,b
658 KJ659650
*,a,b
Tribe Rhynchodemini
Dolichoplana sp. -/
´
Alvarez-Presas, Bagu
˜
n
`
a & Riutort, 2008 DQ665971
a
DQ666037
a,b,d
D. striata Igreginha (Brazil)/Carbayo et al., 2013 KC608341
a
KC608226
a,b,d
425 Bordils (Girona, Spain)/V
*
KJ659698
*,a
KJ659679
*,a,d
660 KJ659683
*,a,d
661 KJ659684
*,a,d
Kontikia ventrolineata 638 Granollers (Barcelona, Spain)/M
*
KJ659681
*,a,d
639 KJ659704
*,a
KJ659682
*,a,d
734 Nursery in Liverpool (UK)/HJ N53.3525 W2.902777
*
KJ659687
*,a,d
739 Saint P
´
ee sur Nivelle (France)/MA N43.34235 W1.52650
*
KJ599732
a
KJ659688
*,a,d
Platydemus manokwari Townsville (Australia)/Bagu
˜
n
`
a et al., 2001 -
´
Alvarez-Presas, Bagu
˜
n
`
a &
Riutort, 2008
DQ665986
a
AF178320
a,bd
Rhynchodemus morph Rs1 411 Vall de’n Bas (Girona, Spain)/P
*
KJ659697
*,a
KJ659676
*,a,d
412 KJ659677
*,a,d
414 KJ659678
*,a,d
(continued on next page)
´
Alvarez-Presas et al. (2014), PeerJ, DOI 10.7717/peerj.430 9/35

Table 2 (continued)
Species/morphotype Code Locality/ref. or Loc code or collector and position GenBank Code
28S COI
908 Luarca, Asturias (Spain)/LL 43
◦
32
′
30.81
′′
N6
◦
32
′
7.42
′′
O
*
KJ659696
*,d
Rhynchodemus cf. sylvaticus Canyamars (Barcelona, Spain)/Mateos et al., 2009 FJ969946
a,bd
091 Canyamars (Barcelona, Spain)/EM N41.598317 E2.44302
*
KJ659672
*,d
219 Montju
¨
ıc (Barcelona, Spain)/Vila-Farr
´
e et al., 2008
*
KJ659673
*,d
569 Underbarrow (UK)/N54.31776 W2.80783
*
KJ659680
*,d
670 Pont en Royans (France)/N45.037875 E5.377033
*
KJ659685
*,d
675 KJ659706
*,a
KJ659686
*,d
905 Benamargosa (M
´
alaga, Spain)/Vila-Farr
´
e et al., 2011
*
KJ659694
*,d
900 Sueiro, Asturias (Spain)/LL N43.527130 W6.877329
*
KJ659689
*,d
901 Aljezur (Portugal)/LL N 37.316146 W8.803392
*
KJ659690
*,d
906 Benamargosa (M
´
alaga, Spain)/Vila-Farr
´
e et al., 2011
*
KJ659695
*,d
Rhynchodemini morph Ri1 902 Benamargosa (M
´
alaga, Spain)/Vila-Farr
´
e et al., 2011
*
KJ659691
*,d
903 Benamargosa (M
´
alaga, Spain)/Vila-Farr
´
e et al., 2011
*
KJ659692
*,d
904 Benamargosa (M
´
alaga, Spain)/Vila-Farr
´
e et al., 2011
*
KJ659693
*,d
Rhynchodemini sp2 262 Int. Park la Amistad, Pila (Panam
´
a)/KA N8.524944 W82.618777
*
KJ659674
*,d
Rhynchodemini sp3 264 KJ659675
*,d
Subfamily Geoplaninae
Cratera crioula S
˜
ao Paulo (Brazil)/Carbayo et al., 2013 KC608441
a
KC608324
a,e
C. tamoia Teres
´
opolis (Brazil)/Carbayo et al., 2013 KC608369
a
KC608254
a,e
Obama sp.6 Bra1 Paulo Lopes (Brazil)/Carbayo et al., 2013 KC608425
a
KC608308
a,e
Bra2 KC608426
a
KC608309
a,e
Obama sp. 434 Bordils (Girona, Spain)/V
*
KJ659652
*,a,e
437 KJ659653
*,a,e
438 KJ659654
*,a,e
667 KJ659663
*,a,e
668 KJ659664
*,a,e
458 Gav
`
a (Barcelona, Spain)/Q
*
KJ659655
*,a,e
459 KJ659656
*,a,e
593 Vilassar de Mar (Barcelona, Spain)/S
*
KJ659657
*,a,e
594 KJ659658
*,a,e
595 KJ659701
*,a
KJ659659
*,a,e
596 La S
´
ınia (Tarragona, Spain)/T
*
KJ659660
*,a,e
610 Treto (Cantabria, Spain)/U
*
KJ659661
*,a,e
611 KJ659662
*,a,e
(continued on next page)
´
Alvarez-Presas et al. (2014), PeerJ, DOI 10.7717/peerj.430 10/35