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<mods:title id="32AE27250305856AFD8053333D0F6591">First record of freshwater planarian Girardia igrina (Platyhelminthes, Tricladida, Continenticola) in Eastern Europe</mods:title>
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<treatment id="6A510E28F463FF90FF18B97DFF048D52" ID-GBIF-Taxon="158635420" LSID="urn:lsid:plazi:treatment:6A510E28F463FF90FF18B97DFF048D52" httpUri="http://treatment.plazi.org/id/6A510E28F463FF90FF18B97DFF048D52" lastPageId="2" lastPageNumber="599" pageId="0" pageNumber="597">
<paragraph id="E247BF3EF463FF92FF18B97DFCA08B59" blockId="0.[151,1437,696,2043]" pageId="0" pageNumber="597">
<subSubSection id="AAE2ECB5F463FF92FF18B97DFDC98CEE" box="[199,531,874,899]" pageId="0" pageNumber="597" type="nomenclature">
<taxonomicName id="25F8C4BDF463FF92FF18B97DFDC98CEE" authority="(Girard, 1850)" baseAuthorityName="Girard" baseAuthorityYear="1850" box="[199,531,874,899]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF463FF92FF18B97DFEAE8CEE" box="[199,372,875,899]" italics="true" pageId="0" pageNumber="597">Girardia tigrina</emphasis>
(
<bibRefCitation id="8669C2CFF463FF92FE5EB97CFDD68CEE" author="Girard, C. F." box="[385,524,874,898]" pageId="0" pageNumber="597" pagination="264 - 265" refId="ref1608" refString="Girard, C. F. (1850) A brief account of the fresh water species of Planariae inhabiting the United States. Proceedings of the Boston Society of Natural History, 3, 264 - 265. [1848 - 1851]" type="journal article" year="1850">Girard, 1850</bibRefCitation>
)
</taxonomicName>
</subSubSection>
<subSubSection id="AAE2ECB5F463FF92FDC6B97CFCA08B59" pageId="0" pageNumber="597" type="distribution">
have been recorded in thirteen countries so far. It is commonly accepted that this species is native to North America, where it is widely distributed, forming a polytypic group (
<bibRefCitation id="8669C2CFF463FF92FB83B998FB0C8CCA" author="Kenk, R." box="[1116,1238,910,934]" pageId="0" pageNumber="597" pagination="1 - 90" refId="ref1874" refString="Kenk, R. (1974) Index of the Genera and Species of the Freshwater Triclads (Turbellaria) of the World. Smithsonian Contributions to Zoology, 183, 1 - 90. https: // doi. org / 10.5479 / si. 00810282.183" type="journal article" year="1974">Kenk 1974</bibRefCitation>
,
<bibRefCitation id="8669C2CFF463FF92FB3EB998FAC38CCA" author="Kenk, R." box="[1249,1305,910,934]" pageId="0" pageNumber="597" pagination="1 - 10" refId="ref1921" refString="Kenk, R. (1989) Revised list of the North American freshwater Planarians (Platyhelminthes: Tricladida: Paludicola). Smithsonian Contributions to Zoology, 476, 1 - 10. https: // doi. org / 10.5479 / si. 00810282.476" type="journal article" year="1989">1989</bibRefCitation>
;
<bibRefCitation id="8669C2CFF463FF92FAFAB998FF158CA6" author="Sluys, R. &amp; Joffe, B. &amp; Cannon, L. R. G." pageId="0" pageNumber="597" pagination="642" refId="ref2189" refString="Sluys, R., Joffe, B. &amp; Cannon, L. R. G. (1995) An alien flatworm in Australian waters. Memoirs of the Queensland Museum, 38, 642." type="journal article" year="1995">
Sluys
<emphasis id="D08C632CF463FF92FAB8B999FA478CCA" box="[1383,1437,910,934]" italics="true" pageId="0" pageNumber="597">et al.</emphasis>
1995
</bibRefCitation>
;
<bibRefCitation id="8669C2CFF463FF92FF05B9A4FE1B8CA6" author="Vila-Farre, M. &amp; Sluys, R. &amp; Almagro, I. &amp; Handberg-Thorsager, M. &amp; Romero, R." box="[218,449,946,970]" pageId="0" pageNumber="597" pagination="1 - 57" refId="ref2466" refString="Vila-Farre, M., Sluys, R., Almagro, I., Handberg-Thorsager, M. &amp; Romero, R. (2011) Freshwater planarians (Platyhelminthes, Tricladida) from the Iberian Peninsula and Greece: diversity and notes on ecology. Zootaxa, 2779 (1), 1 - 57. https: // doi. org / 10.11646 / zootaxa. 2779.1.1" type="journal article" year="2011">
Vila-Farré
<emphasis id="D08C632CF463FF92FE92B9A5FE598CA6" box="[333,387,946,970]" italics="true" pageId="0" pageNumber="597">et al.</emphasis>
2011
</bibRefCitation>
). In the 1920s, it was introduced in Europe (
<bibRefCitation id="8669C2CFF463FF92FC4AB9A4FBE88CA6" author="Meinken, H." box="[917,1074,946,970]" pageId="0" pageNumber="597" pagination="94 - 96" refId="ref2019" refString="Meinken, H. (1925) Ein verkannter Laichrauber. Wochenschrift fur Aquarien und Terrarienkunde, 22, 94 - 96." type="journal article" year="1925">Meinken 1925</bibRefCitation>
) and after that it started to expand southwards and eastwards during the next decades reaching southern
<collectingCountry id="9AEFFFAEF463FF92FC5DB9C0FC178C81" box="[898,973,982,1005]" name="France" pageId="0" pageNumber="597">France</collectingCountry>
,
<collectingCountry id="9AEFFFAEF463FF92FC06B9C0FBCC8C82" box="[985,1046,982,1006]" name="Spain" pageId="0" pageNumber="597">Spain</collectingCountry>
(including the
<collectingRegion id="203C71DCF463FF92FB65B9C0FABD8C82" box="[1210,1383,982,1006]" country="Spain" name="Illes Baleares" pageId="0" pageNumber="597">Balearic Islands</collectingRegion>
) and
<collectingCountry id="9AEFFFAEF463FF92FF48B9EFFF128B7D" box="[151,200,1017,1041]" name="Italy" pageId="0" pageNumber="597">Italy</collectingCountry>
in the late 1960s to early 1970s (
<bibRefCitation id="8669C2CFF463FF92FDE8B9ECFCD18B7E" author="Baguna, J. &amp; Salo, E. &amp; Romero, R." box="[567,779,1017,1042]" pageId="0" pageNumber="597" pagination="15 - 30" refId="ref1508" refString="Baguna, J., Salo, E. &amp; Romero, R. (1980) Les planaries d'aigues dolces a Catalunya i les illes Balears. Butlleti de la Institucio Catalana d'Historia Natural, 45, 15 - 30." type="journal article" year="1980">
Baguñà
<emphasis id="D08C632CF463FF92FD4DB9ECFD108B7D" box="[658,714,1017,1041]" italics="true" pageId="0" pageNumber="597">et al.</emphasis>
1980
</bibRefCitation>
;
<bibRefCitation id="8669C2CFF463FF92FCC8B9EFFC1E8B7D" author="Gourbault, N." box="[791,964,1017,1041]" pageId="0" pageNumber="597" pagination="3 - 7" refId="ref1652" refString="Gourbault, N. (1969) Expansion de Dugesia tigrina (Girard), planaire Americaine introduite en Europe. Annales de Limnologie, 5 (1), 3 - 7. https: // doi. org / 10.1051 / limn / 1969006" type="journal article" year="1969">Gourbault 1969</bibRefCitation>
). There are also records from
<collectingCountry id="9AEFFFAEF463FF92FAD3B9EFFAB48B7D" box="[1292,1390,1017,1041]" name="Australia" pageId="0" pageNumber="597">Australia</collectingCountry>
and
<collectingCountry id="9AEFFFAEF463FF92FF48BE08FF0E8B59" box="[151,212,1054,1077]" name="Japan" pageId="0" pageNumber="597">Japan</collectingCountry>
(
<bibRefCitation id="8669C2CFF463FF92FF3DBE0BFE0F8B59" author="Kawakatsu, M. &amp; Tamura, S. &amp; Takai, M. &amp; Yamamoto, K. &amp; Ueno, R. &amp; Oki, I." box="[226,469,1053,1077]" pageId="0" pageNumber="597" pagination="5 - 11" refId="ref1797" refString="Kawakatsu, M., Tamura, S., Takai, M., Yamamoto, K., Ueno, R. &amp; Oki, I. (1993) The first record of occurence of a naturalized population of Dugesia tigrina (Girard, 1850) at Nagasaki, Kyushu, Japan (Turbellaria, Tricladida, Paludicola). Bulletin of the Biogeographical Society of Japan, 48, 5 - 11." type="journal article" year="1993">
Kawakatsu
<emphasis id="D08C632CF463FF92FEBEBE08FE4C8B59" box="[353,406,1053,1077]" italics="true" pageId="0" pageNumber="597">et al.</emphasis>
1993
</bibRefCitation>
;
<bibRefCitation id="8669C2CFF463FF92FE3FBE0BFD4C8B59" author="Sluys, R. &amp; Joffe, B. &amp; Cannon, L. R. G." box="[480,662,1053,1077]" pageId="0" pageNumber="597" pagination="642" refId="ref2189" refString="Sluys, R., Joffe, B. &amp; Cannon, L. R. G. (1995) An alien flatworm in Australian waters. Memoirs of the Queensland Museum, 38, 642." type="journal article" year="1995">
Sluys
<emphasis id="D08C632CF463FF92FDFDBE08FD828B59" box="[546,600,1053,1077]" italics="true" pageId="0" pageNumber="597">et al.</emphasis>
1995
</bibRefCitation>
;
<bibRefCitation id="8669C2CFF463FF92FD7EBE08FCB48B59" author="Tamura, S. &amp; Oki, I. &amp; Kawakatsu, M. &amp; Ninagawa, M. &amp; Matsusato, T. &amp; Suzuki, H." box="[673,878,1053,1077]" pageId="0" pageNumber="597" pagination="133 - 137" refId="ref2336" refString="Tamura, S., Oki, I., Kawakatsu, M., Ninagawa, M., Matsusato, T. &amp; Suzuki, H. (1985) A note on an introduced species of freshwater planarian, Dugesia tigrina (Girard, 1850), found in culture ponds of australian crayfish in Kagoshima prefecture, Japan. Bulletin of Fuji Women`s College, 23, 133 - 137." type="journal article" year="1985">
Tamura
<emphasis id="D08C632CF463FF92FD26BE08FCF48B59" box="[761,814,1053,1077]" italics="true" pageId="0" pageNumber="597">et al.</emphasis>
1985
</bibRefCitation>
).
</subSubSection>
</paragraph>
<paragraph id="E247BF3EF463FF92FF18BE57FBDA8BCC" blockId="0.[151,1437,696,2043]" pageId="0" pageNumber="597">
In this paper, we present the first record of this species in Eastern Europe. We identified morphologically freshwater planarians found in the Desna River (
<collectingCountry id="9AEFFFAEF463FF92FDEBBE73FD578B11" box="[564,653,1125,1149]" name="Ukraine" pageId="0" pageNumber="597">Ukraine</collectingCountry>
) as
<taxonomicName id="25F8C4BDF463FF92FD62BE73FCB78B10" baseAuthorityName="Girard" baseAuthorityYear="1850" box="[701,877,1125,1149]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF463FF92FD62BE73FCB78B10" box="[701,877,1125,1149]" italics="true" pageId="0" pageNumber="597">Girardia tigrina</emphasis>
</taxonomicName>
. The identity of these planarians was subsequently confirmed by molecular analysis of
<emphasis id="D08C632CF463FF92FDC7BE9EFCDD8BCC" box="[536,775,1160,1184]" italics="true" pageId="0" pageNumber="597">Cytochrome Oxidase I</emphasis>
gene (COI) sequences.
</paragraph>
<subSubSection id="AAE2ECB5F463FF92FF18BEBAFC4F8A1B" pageId="0" pageNumber="597" type="materials_examined">
<paragraph id="E247BF3EF463FF92FF18BEBAFC4F8A1B" blockId="0.[151,1437,696,2043]" pageId="0" pageNumber="597">
Specimens were found in
<materialsCitation id="5290B563F463FF92FE00BEBBFE108A3F" ID-GBIF-Occurrence="2274832805" collectedFrom="with sand and silt at the bottom" collectingDate="2015-06" collectingMethod="collected by hand" collectionCode="Laboratory of the Department of Genetics, Microbiology and Statistics, University of Barcelona, Barcelona, Spain" collectorName="A. V. Martynov" country="Ukraine" latitude="52.18564" location="Desna River (vicinity of Pushkari village, Novhorod-Siverskyi district" longLatPrecision="1" longitude="33.30578" pageId="0" pageNumber="597" specimenCode="MR0472-01, MR0472-11" specimenCount="1" stateProvince="Chernihiv region">
<location id="E727E9E5F463FF92FE00BEBBFB0E8BA8" LSID="urn:lsid:plazi:treatment:6A510E28F463FF90FF18B97DFF048D52:E727E9E5F463FF92FE00BEBBFB0E8BA8" box="[479,1236,1196,1220]" country="Ukraine" latitude="52.18564" longLatPrecision="1" longitude="33.30578" name="Desna River (vicinity of Pushkari village, Novhorod-Siverskyi district" pageId="0" pageNumber="597" stateProvince="Chernihiv region">Desna River (vicinity of Pushkari village, Novhorod-Siverskyi district</location>
,
<collectingRegion id="203C71DCF463FF92FB00BEBAFA428BA8" box="[1247,1432,1196,1220]" country="Ukraine" name="Chernihiv" pageId="0" pageNumber="597">Chernihiv region</collectingRegion>
,
<collectingCountry id="9AEFFFAEF463FF92FF48BEC6FF2A8B84" box="[151,240,1232,1256]" name="Ukraine" pageId="0" pageNumber="597">Ukraine</collectingCountry>
,
<geoCoordinate id="87CCD9F9F463FF92FF25BEC6FE558B84" box="[250,399,1232,1256]" degrees="52" direction="north" minutes="11" orientation="latitude" pageId="0" pageNumber="597" precision="1" seconds="08.3" value="52.18564">52°1108.3”N</geoCoordinate>
<geoCoordinate id="87CCD9F9F463FF92FE4ABEC6FDF68B84" box="[405,556,1232,1256]" degrees="33" direction="east" minutes="18" orientation="longitude" pageId="0" pageNumber="597" precision="1" seconds="20.8" value="33.30578">33°1820.8”E</geoCoordinate>
,
<collectingDate id="86026016F463FF92FDE9BEC6FD7F8B84" box="[566,677,1232,1256]" pageId="0" pageNumber="597" value="2015-06">
<date id="964699FEF463FF92FDE9BEC6FD7F8B84" box="[566,677,1232,1256]" pageId="0" pageNumber="597" value="2015-06">June 2015</date>
</collectingDate>
,
<emphasis id="D08C632CF463FF92FD70BEC6FD048B84" box="[687,734,1232,1256]" italics="true" pageId="0" pageNumber="597">coll.</emphasis>
<collectorName id="4F0DDAE8F463FF92FD3DBEC6FC5F8B84" box="[738,901,1232,1256]" pageId="0" pageNumber="597">A.V. Martynov</collectorName>
),
<collectedFrom id="51E4FCC3F463FF92FC48BEC6FB3C8B84" box="[919,1254,1232,1256]" pageId="0" pageNumber="597">with sand and silt at the bottom</collectedFrom>
. Specimens were
<collectingMethod id="3BB9C729F463FF92FF48BEE2FE8D8A60" box="[151,343,1268,1292]" pageId="0" pageNumber="597">collected by hand</collectingMethod>
. They were preserved in 96% ethanol and are deposited in the collections of the
<collectionCode id="84E927FBF463FF92FB6CBEE2FBAF8A43" pageId="0" pageNumber="597">Laboratory of the Department of Genetics, Microbiology and Statistics, University of Barcelona, Barcelona, Spain</collectionCode>
, under registration numbers
<specimenCode id="B25E1745F463FF92FF48BF2DFEC08A3F" box="[151,282,1339,1363]" pageId="0" pageNumber="597">MR0472-01</specimenCode>
to
<specimenCode id="B25E1745F463FF92FE9EBF2DFE1C8A3F" box="[321,454,1339,1363]" pageId="0" pageNumber="597">MR0472-11</specimenCode>
.
</materialsCitation>
Photographs of fixed animals were taken for identification using camera Leica M165 C. Histological anatomical investigations were not undertaken at this time.
</paragraph>
</subSubSection>
<subSubSection id="AAE2ECB5F463FF90FF18BF95FF048D52" lastPageId="2" lastPageNumber="599" pageId="0" pageNumber="597" type="discussion">
<paragraph id="E247BF3EF463FF92FF18BF95FADE8896" blockId="0.[151,1437,696,2043]" pageId="0" pageNumber="597">
A small piece of the body was taken from the ethanol-fixed individuals for molecular analysis. Genomic DNA was extracted using Wizard Genomic DNA Purification Kit (Promega, Madison, WI,
<collectingCountry id="9AEFFFAEF463FF92FC21BFB0FBE28AD2" box="[1022,1080,1446,1470]" name="United States of America" pageId="0" pageNumber="597">USA</collectingCountry>
). We amplified a fragment of approximately 600 bp from COI gene by polymerase chain reaction (PCR) using the primers BarS (GTTATGCCTGTAAT- GATTG) (
<bibRefCitation id="8669C2CFF463FF92FED9BFFAFDF9896A" author="Alvarez-Presas, M. &amp; Carbayo, F. &amp; Rozas, J. &amp; Riutort, M." box="[262,547,1516,1542]" pageId="0" pageNumber="597" pagination="887 - 896" refId="ref1434" refString="Alvarez-Presas, M., Carbayo, F., Rozas, J. &amp; Riutort, M. (2011) Land planarians (Platyhelminthes) as a model organism for fine-scale phylogeographic studies: understanding patterns of biodiversity in the Brazilian Atlantic Forest hotspot. Journal of Evolutionary Biology, 24, 887 - 896. https: // doi. org / 10.1111 / j. 1420 - 9101.2010.02220. x" type="journal article" year="2011">
Álvarez-Presas
<emphasis id="D08C632CF463FF92FE6FBFF9FE3C896A" box="[432,486,1518,1542]" italics="true" pageId="0" pageNumber="597">et al.</emphasis>
2011
</bibRefCitation>
) and PlatRGi (CATCCTGAGGTTTATATWTTGATT). The PCR reaction was performed in 25 μl, containing 1 μl of DNA, 1× PCR buffer, 2.5 mM MgCl
<subScript id="7E7CBD7BF463FF92FC47BC37FC7A8943" attach="left" box="[920,928,1569,1583]" fontSize="6" pageId="0" pageNumber="597">2</subScript>
, 0.1 mM of each dNTP, 0.5 μM of each primer, and 0.15 units of Taq DNA polymerase (Qiagen). The amplification protocol was: 2 at 95°C, followed by 30 cycles of 94°C for 50, 44°C for 30, 72°C for 50, with a final extension at 72°C for 5. Amplification products were purified using a vacuum manifold (Multi-Well Plate Vacuum Manifold, Pall Corporation, MI,
<collectingCountry id="9AEFFFAEF463FF92FC2BBC6BFBF189F9" box="[1012,1067,1661,1685]" name="United States of America" pageId="0" pageNumber="597">USA</collectingCountry>
) and sequenced in both directions on a DNA Analyzer 96-capillary sequencer at Macrogen Inc. (Amsterdam,
<collectingCountry id="9AEFFFAEF463FF92FC61BCB6FBAF89D4" box="[958,1141,1696,1720]" name="Netherlands" pageId="0" pageNumber="597">The Netherlands</collectingCountry>
). A contig was obtained using Genious
<date id="964699FEF463FF92FEFDBCD3FE8D89B0" box="[290,343,1732,1756]" pageId="0" pageNumber="597" value="2010-05">v. 10</date>
(https://www.geneious.com), the resulting sequence had been deposited in GenBank under accession number
<accessionNumber id="FDAB22DDF463FF92FF2EBCFEFEB5886C" box="[241,367,1768,1792]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/MN092348" pageId="0" pageNumber="597">
MN
<date id="964699FEF463FF92FEC2BCFEFEB5886C" box="[285,367,1768,1792]" pageId="0" pageNumber="597" value="48-09-23">092348</date>
</accessionNumber>
. The sequence obtained was compared to the GenBank database content using nBLAST. The best hits, with similarities between 89-90% and e-values &lt;
<date id="964699FEF463FF92FD09BD1AFD2D884F" box="[726,759,1804,1827]" pageId="0" pageNumber="597">7 x</date>
e
<superScript id="158D1276F463FF92FCD0BD1EFCF6887B" attach="right" box="[783,812,1800,1815]" fontSize="6" pageId="0" pageNumber="597">-159</superScript>
, were to three
<taxonomicName id="25F8C4BDF463FF92FC17BD1AFBFC8848" authorityName="Ball" authorityYear="1974" box="[968,1062,1804,1828]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="genus">
<emphasis id="D08C632CF463FF92FC17BD1AFBFC8848" box="[968,1062,1804,1828]" italics="true" pageId="0" pageNumber="597">Girardia</emphasis>
</taxonomicName>
species (
<taxonomicName id="25F8C4BDF463FF92FB57BD1AFB26884F" authorityName="Chen &amp; Wang" authorityYear="2015" box="[1160,1276,1804,1827]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="sinensis">
<emphasis id="D08C632CF463FF92FB57BD1AFB26884F" box="[1160,1276,1804,1827]" italics="true" pageId="0" pageNumber="597">G. sinensis</emphasis>
</taxonomicName>
<accessionNumber id="FDAB22DDF463FF92FADCBD1AFAA28848" box="[1283,1400,1804,1828]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/KP091892" pageId="0" pageNumber="597">
KP
<date id="964699FEF463FF92FAFABD1AFAA28848" box="[1317,1400,1804,1828]" pageId="0" pageNumber="597" value="92-09-18">091892</date>
</accessionNumber>
;
<taxonomicName id="25F8C4BDF463FF92FA5DBD1AFF3A882B" baseAuthorityName="Girard" baseAuthorityYear="1850" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF463FF92FA5DBD1AFF3A882B" italics="true" pageId="0" pageNumber="597">G. tigrina</emphasis>
</taxonomicName>
<accessionNumber id="FDAB22DDF463FF92FF3ABD39FEBA882B" box="[229,352,1839,1863]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/KM200930" pageId="0" pageNumber="597">
KM
<date id="964699FEF463FF92FECFBD39FEBA882B" box="[272,352,1839,1863]" pageId="0" pageNumber="597" value="30-09-20">200930</date>
</accessionNumber>
and
<taxonomicName id="25F8C4BDF463FF92FE4EBD26FD96882B" baseAuthorityName="Woodworth" baseAuthorityYear="1897" box="[401,588,1839,1863]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="dorotocephala">
<emphasis id="D08C632CF463FF92FE4EBD26FD96882B" box="[401,588,1839,1863]" italics="true" pageId="0" pageNumber="597">G. dorotocephala</emphasis>
</taxonomicName>
<accessionNumber id="FDAB22DDF463FF92FD8EBD39FD0B882B" box="[593,721,1839,1863]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/KM200929" pageId="0" pageNumber="597">
KM
<date id="964699FEF463FF92FDA2BD39FD0B882B" box="[637,721,1839,1863]" pageId="0" pageNumber="597" value="29-09-20">200929</date>
</accessionNumber>
). We then downloaded the COI sequences from these three
<taxonomicName id="25F8C4BDF463FF92FAE1BD39FA46882B" authorityName="Ball" authorityYear="1974" box="[1342,1436,1839,1863]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="genus">
<emphasis id="D08C632CF463FF92FAE1BD39FA46882B" box="[1342,1436,1839,1863]" italics="true" pageId="0" pageNumber="597">Girardia</emphasis>
</taxonomicName>
species from GenBank plus that of
<taxonomicName id="25F8C4BDF463FF92FDCCBD42FD408807" baseAuthorityName="Marcus" baseAuthorityYear="1946" box="[531,666,1875,1899]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="schubarti">
<emphasis id="D08C632CF463FF92FDCCBD42FD408807" box="[531,666,1875,1899]" italics="true" pageId="0" pageNumber="597">G. schubarti</emphasis>
</taxonomicName>
(
<accessionNumber id="FDAB22DDF463FF92FD77BD45FCF88807" box="[680,802,1875,1899]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/DQ666041" pageId="0" pageNumber="597">DQ666041</accessionNumber>
) and obtained an alignment including as well sequences of two
<taxonomicName id="25F8C4BDF463FF92FF18BD61FEFF88E3" authorityName="Ball" authorityYear="1974" box="[199,293,1911,1935]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="genus">
<emphasis id="D08C632CF463FF92FF18BD61FEFF88E3" box="[199,293,1911,1935]" italics="true" pageId="0" pageNumber="597">Girardia</emphasis>
</taxonomicName>
sister genera, two
<taxonomicName id="25F8C4BDF463FF92FE2DBD6EFD9188E3" authorityName="Girard" authorityYear="1850" box="[498,587,1912,1935]" class="Rhabditophora" family="Dugesiidae" genus="Dugesia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="genus">
<emphasis id="D08C632CF463FF92FE2DBD6EFD9188E3" box="[498,587,1912,1935]" italics="true" pageId="0" pageNumber="597">Dugesia</emphasis>
</taxonomicName>
(
<taxonomicName id="25F8C4BDF463FF92FD83BD6EFD3388E3" authorityName="Stocchino &amp; Sluys" authorityYear="2017" box="[604,745,1911,1935]" class="Rhabditophora" family="Dugesiidae" genus="Dugesia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="sigmoides">
<emphasis id="D08C632CF463FF92FD83BD6EFD3388E3" box="[604,745,1911,1935]" italics="true" pageId="0" pageNumber="597">D. sigmoides</emphasis>
</taxonomicName>
<accessionNumber id="FDAB22DDF463FF92FD2FBD61FCBD88E3" box="[752,871,1911,1935]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/KY498849" pageId="0" pageNumber="597">KY498849</accessionNumber>
and
<taxonomicName id="25F8C4BDF463FF92FC42BD6EFB9388E3" baseAuthorityName="Duges" baseAuthorityYear="1830" box="[925,1097,1911,1935]" class="Rhabditophora" family="Dugesiidae" genus="Dugesia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="gonocephala">
<emphasis id="D08C632CF463FF92FC42BD6EFB9388E3" box="[925,1097,1911,1935]" italics="true" pageId="0" pageNumber="597">D. gonocephala</emphasis>
</taxonomicName>
<accessionNumber id="FDAB22DDF463FF92FB8EBD61FB6188E3" box="[1105,1211,1911,1935]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/FJ646985" pageId="0" pageNumber="597">FJ646985</accessionNumber>
) and four
<taxonomicName id="25F8C4BDF463FF92FAF2BD61FA4688E3" authorityName="Ball" authorityYear="1974" box="[1325,1436,1911,1935]" class="Rhabditophora" family="Dugesiidae" genus="Schmidtea" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="genus">
<emphasis id="D08C632CF463FF92FAF2BD61FA4688E3" box="[1325,1436,1911,1935]" italics="true" pageId="0" pageNumber="597">Schmidtea</emphasis>
</taxonomicName>
(
<accessionNumber id="FDAB22DDF463FF92FF40BD8DFEC488DF" box="[159,286,1947,1971]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/KM821047" pageId="0" pageNumber="597">KM821047</accessionNumber>
,
<accessionNumber id="FDAB22DDF463FF92FEF5BD8CFE4988DE" box="[298,403,1946,1970]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/JF837062" pageId="0" pageNumber="597">JF837062</accessionNumber>
<emphasis id="D08C632CF463FF92FE44BD8DFD8788DF" box="[411,605,1947,1971]" italics="true" pageId="0" pageNumber="597">S. mediterranea-1</emphasis>
and
<emphasis id="D08C632CF463FF92FD4BBD8DFD7B88DF" box="[660,673,1947,1971]" italics="true" pageId="0" pageNumber="597">2</emphasis>
respecively;
<accessionNumber id="FDAB22DDF463FF92FCEDBD8CFC7788DE" box="[818,941,1946,1970]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/MG457275" pageId="0" pageNumber="597">MG457275</accessionNumber>
<taxonomicName id="25F8C4BDF463FF92FC6ABD8DFBF088DF" authorityName="Schmidt" authorityYear="1861" box="[949,1066,1947,1971]" class="Rhabditophora" family="Dugesiidae" genus="Schmidtea" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="lugubris">
<emphasis id="D08C632CF463FF92FC6ABD8DFBF088DF" box="[949,1066,1947,1971]" italics="true" pageId="0" pageNumber="597">S. lugubris</emphasis>
</taxonomicName>
;
<accessionNumber id="FDAB22DDF463FF92FBE5BD8CFB6F88DE" box="[1082,1205,1946,1970]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/MG457277" pageId="0" pageNumber="597">MG457277</accessionNumber>
<emphasis id="D08C632CF463FF92FB62BD8DFAD788DE" box="[1213,1293,1947,1970]" italics="true" pageId="0" pageNumber="597">S. nova</emphasis>
). Finally, we included COI sequences of two Planarioidea (
<taxonomicName id="25F8C4BDF463FF92FD50BDA9FC8288BA" authorityName="Hyman" authorityYear="1931" box="[655,856,1983,2007]" class="Rhabditophora" family="Planariidae" genus="Polycelis" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="coronata">
<emphasis id="D08C632CF463FF92FD50BDA9FC8288BA" box="[655,856,1983,2007]" italics="true" pageId="0" pageNumber="597">Polycelis coronata</emphasis>
</taxonomicName>
<accessionNumber id="FDAB22DDF463FF92FCBEBDA8FC0288BA" box="[865,984,1982,2006]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/KY057186" pageId="0" pageNumber="597">KY057186</accessionNumber>
and
<taxonomicName id="25F8C4BDF463FF92FBCFBDA9FB1B88BB" baseAuthorityName="Dana" baseAuthorityYear="1766" box="[1040,1217,1983,2007]" class="Rhabditophora" family="Planariidae" genus="Crenobia" kingdom="Animalia" order="Seriata" pageId="0" pageNumber="597" phylum="Platyhelminthes" rank="species" species="alpina">
<emphasis id="D08C632CF463FF92FBCFBDA9FB1B88BB" box="[1040,1217,1983,2007]" italics="true" pageId="0" pageNumber="597">Crenobia alpina</emphasis>
</taxonomicName>
<accessionNumber id="FDAB22DDF463FF92FB15BDA8FA9888BA" box="[1226,1346,1982,2006]" httpUri="https://www.ebi.ac.uk/ena/browser/api/embl/KY569153" pageId="0" pageNumber="597">KY569153</accessionNumber>
) as outgroups. We inferred a phylogenetic tree by maximum likelihood using RAxML v 7.2.6 (
<bibRefCitation id="8669C2CFF463FF92FB9EBDF4FB238896" author="Stamatakis, A." box="[1089,1273,2018,2042]" pageId="0" pageNumber="597" pagination="2688 - 2690" refId="ref2294" refString="Stamatakis, A. (2006) RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics, 22, 2688 - 2690. https: // doi. org / 10.1093 / bioinformatics / btl 446" type="journal article" year="2006">Stamatakis 2006</bibRefCitation>
).
</paragraph>
<caption id="B687EFB6F462FF93FF48BEB5FE7B8BB0" httpUri="https://zenodo.org/record/3335850/files/figure.png" pageId="1" pageNumber="598" startId="1.[151,244,1187,1209]" targetBox="[157,1430,194,1166]" targetPageId="1">
<paragraph id="E247BF3EF462FF93FF48BEB5FE7B8BB0" pageId="1" pageNumber="598">
<emphasis id="D08C632CF462FF93FF48BEB5FED78BD4" bold="true" box="[151,269,1187,1209]" pageId="1" pageNumber="598">FIGURE 1.</emphasis>
Maximum-Likelihood tree of COI gene, the scale indicates the branch length in substitutions per site. Numbers at nodes indicate bootstrap supports.
</paragraph>
</caption>
<paragraph id="E247BF3EF462FF90FF18BF1BFE168E72" blockId="1.[151,1437,1293,2037]" lastBlockId="2.[151,1437,154,574]" lastPageId="2" lastPageNumber="599" pageId="1" pageNumber="598">
The phylogenetic tree (
<figureCitation id="7AC3A3BBF462FF93FE60BF1BFE248A49" box="[447,510,1293,1317]" captionStart="FIGURE 1" captionStartId="1.[151,244,1187,1209]" captionTargetBox="[157,1430,194,1166]" captionTargetId="graphics@1.[157,1318,194,1103]" captionTargetPageId="1" captionText="FIGURE 1. Maximum-Likelihood tree of COI gene, the scale indicates the branch length in substitutions per site. Numbers at nodes indicate bootstrap supports." httpUri="https://zenodo.org/record/3335850/files/figure.png" pageId="1" pageNumber="598">Fig. 1</figureCitation>
) shows that the sequence obtained in the present work forms a highly supported clade with
<taxonomicName id="25F8C4BDF462FF93FF0FBF24FEE78A25" baseAuthorityName="Girard" baseAuthorityYear="1850" box="[208,317,1330,1353]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF462FF93FF0FBF24FEE78A25" box="[208,317,1330,1353]" italics="true" pageId="1" pageNumber="598">G. tigrina</emphasis>
</taxonomicName>
,
<taxonomicName id="25F8C4BDF462FF93FE92BF24FDD18A25" baseAuthorityName="Woodworth" baseAuthorityYear="1897" box="[333,523,1329,1353]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="dorotocephala">
<emphasis id="D08C632CF462FF93FE92BF24FDD18A25" box="[333,523,1329,1353]" italics="true" pageId="1" pageNumber="598">G. dorotocephala</emphasis>
</taxonomicName>
and
<taxonomicName id="25F8C4BDF462FF93FD9ABF24FD668A25" authorityName="Chen &amp; Wang" authorityYear="2015" box="[581,700,1330,1353]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="sinensis">
<emphasis id="D08C632CF462FF93FD9ABF24FD668A25" box="[581,700,1330,1353]" italics="true" pageId="1" pageNumber="598">G. sinensis</emphasis>
</taxonomicName>
, that together constitute the sister group of
<taxonomicName id="25F8C4BDF462FF93FB7ABF24FAF78A25" baseAuthorityName="Marcus" baseAuthorityYear="1946" box="[1189,1325,1329,1353]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="schubarti">
<emphasis id="D08C632CF462FF93FB7ABF24FAF78A25" box="[1189,1325,1329,1353]" italics="true" pageId="1" pageNumber="598">G. schubarti</emphasis>
</taxonomicName>
. The new sequence hence clearly belongs to the genus
<taxonomicName id="25F8C4BDF462FF93FD5BBF43FD388A01" authorityName="Ball" authorityYear="1974" box="[644,738,1365,1389]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="genus">
<emphasis id="D08C632CF462FF93FD5BBF43FD388A01" box="[644,738,1365,1389]" italics="true" pageId="1" pageNumber="598">Girardia</emphasis>
</taxonomicName>
, it does not show a closest relationship to any of the three species, probably due to the variability of the COI gene. Having only one sequence per species does not allow to have all the variability of the gene within each species represented and, on the other hand, the three species are quite similar for their COI (compare the short branches separating
<taxonomicName id="25F8C4BDF462FF93FD6EBFD4FCFC8AB5" authorityName="Chen &amp; Wang" authorityYear="2015" box="[689,806,1474,1497]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="sinensis">
<emphasis id="D08C632CF462FF93FD6EBFD4FCFC8AB5" box="[689,806,1474,1497]" italics="true" pageId="1" pageNumber="598">G. sinensis</emphasis>
</taxonomicName>
and
<taxonomicName id="25F8C4BDF462FF93FC83BFD4FC1C8AB5" baseAuthorityName="Girard" baseAuthorityYear="1850" box="[860,966,1474,1497]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF462FF93FC83BFD4FC1C8AB5" box="[860,966,1474,1497]" italics="true" pageId="1" pageNumber="598">G. tigrina</emphasis>
</taxonomicName>
with the ones separating two individuals of
<taxonomicName id="25F8C4BDF462FF93FF48BFF0FE988A91" box="[151,322,1509,1533]" class="Rhabditophora" family="Dugesiidae" genus="Schmidtea" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="mediterranea">
<emphasis id="D08C632CF462FF93FF48BFF0FE988A91" box="[151,322,1509,1533]" italics="true" pageId="1" pageNumber="598">S. mediterranea</emphasis>
</taxonomicName>
). Although
<taxonomicName id="25F8C4BDF462FF93FE1BBFF0FDE38A91" authorityName="Chen &amp; Wang" authorityYear="2015" box="[452,569,1510,1533]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="sinensis">
<emphasis id="D08C632CF462FF93FE1BBFF0FDE38A91" box="[452,569,1510,1533]" italics="true" pageId="1" pageNumber="598">G. sinensis</emphasis>
</taxonomicName>
was described from
<collectingCountry id="9AEFFFAEF462FF93FCC5BFF3FC808A91" box="[794,858,1509,1533]" name="China" pageId="1" pageNumber="598">China</collectingCountry>
(
<bibRefCitation id="8669C2CFF462FF93FCB7BFF3FBC58A91" author="Chen, Y. - H. &amp; Chen, X. - M. &amp; Wu, C. - C. &amp; Wang, A. - T." box="[872,1055,1509,1533]" pageId="1" pageNumber="598" pagination="166 - 178" refId="ref1550" refString="Chen, Y. - H., Chen, X. - M., Wu, C. - C. &amp; Wang, A. - T. (2015) A new species of the genus Girardia (Tricladida: Dugesiidae) from China. Zoological Systematics, 40, 166 - 178." type="journal article" year="2015">
Chen
<emphasis id="D08C632CF462FF93FC76BFF0FC3A8A91" box="[937,992,1509,1533]" italics="true" pageId="1" pageNumber="598">et al.</emphasis>
2015
</bibRefCitation>
), its close relationship to
<taxonomicName id="25F8C4BDF462FF93FAEDBFF0FA478A91" baseAuthorityName="Girard" baseAuthorityYear="1850" box="[1330,1437,1510,1533]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF462FF93FAEDBFF0FA478A91" box="[1330,1437,1510,1533]" italics="true" pageId="1" pageNumber="598">G. tigrina</emphasis>
</taxonomicName>
and
<taxonomicName id="25F8C4BDF462FF93FF1BBC1CFEA5894D" baseAuthorityName="Woodworth" baseAuthorityYear="1897" box="[196,383,1545,1569]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="dorotocephala">
<emphasis id="D08C632CF462FF93FF1BBC1CFEA5894D" box="[196,383,1545,1569]" italics="true" pageId="1" pageNumber="598">G. dorotocephala</emphasis>
</taxonomicName>
in the phylogenetic tree most probably indicates that it is also a case of a recent introduction in that country coming from North America. Sequences from more individuals of all these species will be needed to molecularly identify the sequence we obtained from the Ukrainian specimen, and also to clarify the relationships among the three species and even their species status. From their external appearance nonetheless, the specimens (
<figureCitation id="7AC3A3BBF462FF93FB75BC63FB3089E1" box="[1194,1258,1653,1677]" captionStart="FIGURE 2" captionStartId="2.[151,244,1764,1786]" captionTargetBox="[151,1435,597,1744]" captionTargetId="figure@2.[151,1435,597,1744]" captionText="FIGURE 2. A preserved specimen of Girardia tigrina (Girard, 1850) collected from Desna River near Pushkari village (Ukraine). A. Ventral view. B. Anterior part in dorsal view, posterior part in ventral view. C. Dorsal view. ph—pharynx, m—mouth, e—eye." httpUri="https://zenodo.org/record/3335852/files/figure.png" pageId="1" pageNumber="598">Fig. 2</figureCitation>
) collected in the studied locality fall within
<typeStatus id="3D43019CF462FF93FE64BC8CFE3389DE" box="[443,489,1690,1714]" pageId="1" pageNumber="598">type</typeStatus>
I (sensu
<bibRefCitation id="8669C2CFF462FF93FD92BC8CFD0089DD" author="Hyman, L. H." box="[589,730,1689,1714]" pageId="1" pageNumber="598" pagination="419 - 439" refId="ref1698" refString="Hyman, L. H. (1939) New species of flatworms from North, Central, and South America. Proceedings of the United States National Museum, 86, 419 - 439. https: // doi. org / 10.5479 / si. 00963801.86 - 3055.419" type="journal article" year="1939">Hyman 1939</bibRefCitation>
) and class A of
<taxonomicName id="25F8C4BDF462FF93FC56BC8FFBED89DD" baseAuthorityName="Girard" baseAuthorityYear="1850" box="[905,1079,1689,1713]" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF462FF93FC56BC8FFBED89DD" box="[905,1079,1689,1713]" italics="true" pageId="1" pageNumber="598">Girardia tigrina</emphasis>
</taxonomicName>
according to
<bibRefCitation id="8669C2CFF462FF93FB13BC8FFA4389DD" author="Ribas, M. &amp; Riutort, M. &amp; Baguna, J." box="[1228,1433,1689,1713]" pageId="1" pageNumber="598" pagination="609 - 626" refId="ref2042" refString="Ribas, M., Riutort, M. &amp; Baguna, J. (1989) Morphological and biochemical variation in populations of Dugesia (G.) tigrina (Turbellaria, Tricladida, Paludicola) from the western Mediterranean: biogeographical and taxonomical implications. Journal of Zoology, 218, 609 - 626. https: // doi. org / 10.1111 / j. 1469 - 7998.1989. tb 05003. x" type="journal article" year="1989">
Ribas
<emphasis id="D08C632CF462FF93FACFBC8CFA9D89DD" box="[1296,1351,1689,1713]" italics="true" pageId="1" pageNumber="598">et al.</emphasis>
(1989)
</bibRefCitation>
. These are fissiparous specimens, externally spotted (numerous small black spots intermingled with bigger white spots) on a brownish background and with a pharynx clearly pigmented. The previous reports on distributions of European populations show that the
<typeStatus id="3D43019CF462FF93FEB1BD10FE468872" box="[366,412,1798,1822]" pageId="1" pageNumber="598">type</typeStatus>
I-class A populations are the commonest
<typeStatus id="3D43019CF462FF93FCB8BD10FC4F8872" box="[871,917,1798,1822]" pageId="1" pageNumber="598">type</typeStatus>
found. These fissiparous populations are mixoploids, presenting diploid (2n = 16) and triploid (3n = 24) cells, while the sexuals are always diploids (
<bibRefCitation id="8669C2CFF462FF93FB08BD3FFA4A882D" author="Ribas, M. &amp; Riutort, M. &amp; Baguna, J." box="[1239,1424,1833,1857]" pageId="1" pageNumber="598" pagination="609 - 626" refId="ref2042" refString="Ribas, M., Riutort, M. &amp; Baguna, J. (1989) Morphological and biochemical variation in populations of Dugesia (G.) tigrina (Turbellaria, Tricladida, Paludicola) from the western Mediterranean: biogeographical and taxonomical implications. Journal of Zoology, 218, 609 - 626. https: // doi. org / 10.1111 / j. 1469 - 7998.1989. tb 05003. x" type="journal article" year="1989">
Ribas
<emphasis id="D08C632CF462FF93FAC3BD3CFA8B882D" box="[1308,1361,1833,1857]" italics="true" pageId="1" pageNumber="598">et al.</emphasis>
1989
</bibRefCitation>
). After studying the populations of the western Mediterranean,
<bibRefCitation id="8669C2CFF462FF93FCF0BD5BFC228809" author="Ribas, M. &amp; Riutort, M. &amp; Baguna, J." box="[815,1016,1869,1893]" pageId="1" pageNumber="598" pagination="609 - 626" refId="ref2042" refString="Ribas, M., Riutort, M. &amp; Baguna, J. (1989) Morphological and biochemical variation in populations of Dugesia (G.) tigrina (Turbellaria, Tricladida, Paludicola) from the western Mediterranean: biogeographical and taxonomical implications. Journal of Zoology, 218, 609 - 626. https: // doi. org / 10.1111 / j. 1469 - 7998.1989. tb 05003. x" type="journal article" year="1989">
Ribas
<emphasis id="D08C632CF462FF93FCACBD58FC708809" box="[883,938,1869,1893]" italics="true" pageId="1" pageNumber="598">et al.</emphasis>
(1989)
</bibRefCitation>
suggested that several introductions of different stocks, races, or subspecies, living within the area of origin, have occurred since populations in Europe differ from the commonly described fissiparous population both in pigmentation patterns and reproductive biology.
<taxonomicName id="25F8C4BDF462FF93FAE1BD83FF3A88BD" baseAuthorityName="Girard" baseAuthorityYear="1850" class="Rhabditophora" family="Dugesiidae" genus="Girardia" kingdom="Animalia" order="Seriata" pageId="1" pageNumber="598" phylum="Platyhelminthes" rank="species" species="tigrina">
<emphasis id="D08C632CF462FF93FAE1BD83FF3A88BD" italics="true" pageId="1" pageNumber="598">Girardia tigrina</emphasis>
</taxonomicName>
has also been recorded from
<collectingCountry id="9AEFFFAEF462FF93FDC1BDAFFDB888BD" box="[542,610,1977,2001]" name="Brazil" pageId="1" pageNumber="598">Brazil</collectingCountry>
,
<bibRefCitation id="8669C2CFF462FF93FDB2BDACFCD688BD" author="Marcus, E." box="[621,780,1977,2001]" pageId="1" pageNumber="598" pagination="5 - 254" refId="ref1967" refString="Marcus, E. (1946) Sobre Turbellaria Brasileiros. Boletins da Faculdade de Philosophia, Sciencias e Letras, Universidade de Sao Paulo. Zoologia, 11, 5 - 254. https: // doi. org / 10.11606 / issn. 2526 - 4877. bsffclzoologia. 1946.125301" type="journal article" year="1946">Marcus (1946)</bibRefCitation>
considered the species to be indigenous in
<collectingCountry id="9AEFFFAEF462FF93FB3FBDAFFAFB88BD" box="[1248,1313,1977,2001]" name="Brazil" pageId="1" pageNumber="598">Brazil</collectingCountry>
rather than descended from North America.
<bibRefCitation id="8669C2CFF462FF93FE28BDCBFC4B8899" author="Kawakatsu, M. &amp; Ponce de Leon, R." box="[503,913,2013,2037]" pageId="1" pageNumber="598" pagination="5 - 14" refId="ref1748" refString="Kawakatsu, M. &amp; Ponce de Leon, R. (1990) The occurrence of Dugesia tigrina (Girard, 1850) (Turbellaria: Tricladida: Paludicola) in Uruguay. Proceedings of the Japanese Society of Systematic Zoology, 41, 5 - 14." type="journal article" year="1990">Kawakatsu and Ponce de León (1990)</bibRefCitation>
did not exclude the possibility that the Brazilian populations of the species were introduced from North America. Finally, Sluys
<emphasis id="D08C632CF461FF90FC24BA8DFBE98FDE" box="[1019,1075,154,178]" italics="true" pageId="2" pageNumber="599">et al.</emphasis>
(2005) pointed slight anatomical differences between North and South American specimens of
<emphasis id="D08C632CF461FF90FCF5BAA8FC498FB9" box="[810,915,190,213]" italics="true" pageId="2" pageNumber="599">G. tigrina</emphasis>
and suggested that it might well indicate that the North and South American forms concerned sibling species. Though, they refrained from coining a new species name for the South American animals.
</paragraph>
<paragraph id="E247BF3EF461FF90FF18BB3CFF048D52" blockId="2.[151,1437,154,574]" pageId="2" pageNumber="599">
In conclusion, we report the first case of a
<emphasis id="D08C632CF461FF90FD47BB3CFD2C8E2E" box="[664,758,298,322]" italics="true" pageId="2" pageNumber="599">Girardia</emphasis>
species,
<emphasis id="D08C632CF461FF90FC86BB3CFC1E8E2D" box="[857,964,298,321]" italics="true" pageId="2" pageNumber="599">G. tigrina</emphasis>
, in Ukraine and Eastern Europe. Moreover, our phylogenetic results showing the tight relationship between
<emphasis id="D08C632CF461FF90FC8FBB58FC668E09" box="[848,956,334,357]" italics="true" pageId="2" pageNumber="599">G. tigrina</emphasis>
,
<emphasis id="D08C632CF461FF90FC14BB58FB528E0A" box="[971,1160,334,358]" italics="true" pageId="2" pageNumber="599">G. dorotocephala</emphasis>
and
<emphasis id="D08C632CF461FF90FB60BB58FAEF8E09" box="[1215,1333,334,357]" italics="true" pageId="2" pageNumber="599">G. sinensis</emphasis>
, together with the existing literature on these species, point to the need of a thorough revision for the whole genus. This situation derives from a poor sampling and systematic study of
<emphasis id="D08C632CF461FF90FD05BB80FCE28EC2" box="[730,824,406,430]" italics="true" pageId="2" pageNumber="599">Girardia</emphasis>
in its area of origin, America. As well, whether single, or multiple and independent introductions of
<emphasis id="D08C632CF461FF90FDBFBBACFD648EBE" box="[608,702,442,466]" italics="true" pageId="2" pageNumber="599">Girardia</emphasis>
have occurred in Europe is still an open question. These riddles can only be solved by a comprehensive comparison of distantly located European populations to the original lineages from North America. Therefore, more sampling efforts and an in-depth systematic study should be conducted to clarify all these issues.
</paragraph>
</subSubSection>
</treatment>
</document>