@article {1902, title = {Hidden diversity in forest soils: Characterization and comparison of terrestrial flatworm{\textquoteright}s communities in two national parks in Spain}, journal = {Ecology and EvolutionEcology and EvolutionEcol Evol}, volume = {8}, year = {2018}, note = {doi: 10.1002/ece3.4178}, month = {2018/08/01}, pages = {7386 - 7400}, publisher = {John Wiley \& Sons, Ltd}, abstract = {Abstract Terrestrial flatworms (Platyhelminthes, Tricladida, and Geoplanidae) belong to what is known as cryptic soil fauna of humid forests and are animals not easily found or captured in traps. Nonetheless, they have been demonstrated to be good indicators of the conservation status of their habitat as well as a good model to reconstruct the recent and old events affecting biodiversity. This is mainly due to their delicate constitution, their dependence on the integrity of their habitat, and their very low dispersal capacity. At present, little is known about their communities, except for some studies performed in Brazil. In this work, we analyze for the first time in Europe terrestrial flatworm communities. We have selected two protected areas belonging to the Red Espa{\~n}ola de Parques Nacionales. Our aims include performing a first study of the species richness and community structure for European terrestrial planarian species at regional and local scale. We evaluate the effect of type of forests in the community composition and flatworms? abundance, but also have into account the phylogenetic framework (never considered in previous studies) analyzed based on molecular data. We find differences in the species composition among parks, with an astonishingly high diversity of endemic species in the Parque Nacional de Picos de Europa and an extremely low diversity of species in the Parque Nacional de Ordesa y Monte Perdido. These divergent patterns cannot be attributed to differences in physical variables, and in addition, the analyses of their phylogenetic relationships and, for a few species, their genetic structure, point to a more probable historical explanation.}, keywords = {Last Glacial Maximum, Microplana, molecular phylogenetics, refugia, soil ecology, species diversity}, isbn = {2045-7758}, url = {https://doi.org/10.1002/ece3.4178}, author = {{\'A}lvarez-Presas, Marta and Mateos, Eduardo and Riutort, Marta} } @inbook {1901, title = {Planarian Diversity and Phylogeny}, booktitle = {Planarian Regeneration: Methods and Protocols}, year = {2018}, month = {2018//}, pages = {1 - 56}, publisher = {Springer New York}, organization = {Springer New York}, address = {New York, NY}, abstract = {Hundreds of planarian species exist worldwide, representing a rich phenotypic diversity. This chapter presents an overview of the morphology and anatomy of various taxonomic groups of planarian flatworms, focusing on features enabling recognition and identification of the animals. The most recent view on the phylogenetic relationships of the planarians is presented, together with geographic distribution patterns of major groups of triclads. The chapter concludes with a brief methodological section outlining species identification on basis of anatomical features. In conjunction with the established laboratory model species, the phenotypic diversity of planarians provides rich opportunities for comparative studies, which this chapter aims to inspire.}, isbn = {978-1-4939-7802-1}, url = {https://doi.org/10.1007/978-1-4939-7802-1_1}, author = {Sluys, Ronald and Riutort, Marta}, editor = {Rink, Jochen C.} } @article {doi:10.1111/jzs.12214, title = {Diversification and biogeographic history of the Western Palearctic freshwater flatworm genus Schmidtea (Tricladida: Dugesiidae), with a redescription of Schmidtea nova}, journal = {Journal of Zoological Systematics and Evolutionary Research}, volume = {56}, number = {3}, year = {2018}, pages = {335-351}, abstract = {Abstract The freshwater flatworm genus Schmidtea is endemic in the Western Palearctic region, where it is represented by only four species, thus contrasting with the high species diversity of the closely related genus Dugesia within Europe. Although containing an important model species in developmental and regeneration research, viz. Schmidtea mediterranea, no evolutionary studies on the genus Schmidtea have been undertaken. For the first time, we present a well-resolved molecular phylogenetic tree of the four species of the genus, inferred on the basis of two molecular markers, and provide also the first detailed morphological account of Schmidtea nova. The phylogenetic tree generated corroborates an earlier speciation hypothesis based on karyological data and points to chromosomal rearrangements as the main drivers of speciation in this genus. The high genetic divergence between the four species, in combination with previous dating studies and their current geographic distribution, suggests that Schmidtea could have originated in Laurasia but lost most of its diversity during the Oligocene. Thus, its present distribution pattern may be the result of the expansion of three of its four relictual species over Europe, probably after the Pleistocene glaciations. Our detailed morphological study of S.~nova revealed that it shows a number of remarkable features: interconnected testis follicles, parovaria, an ejaculatory duct exiting into the primary as well as the secondary seminal vesicle by means of a nipple, and the wall of the distal section of the ejaculatory duct being sclerotic or chitinized.}, keywords = {Biogeography, molecular phylogeny, Schmidtea, speciation, taxonomy}, doi = {10.1111/jzs.12214}, url = {https://onlinelibrary.wiley.com/doi/abs/10.1111/jzs.12214}, author = {Leria, Laia and Sluys, Ronald and Riutort, Marta} } @article {1698, title = {First report of the land planarian Diversibipalium multilineatum (Makino \& Shirasawa, 1983) (Platyhelminthes, Tricladida, Continenticola) in Europe.}, journal = {Zootaxa}, volume = {4067}, year = {2016}, month = {2016 Jan 26}, pages = {577-80}, abstract = {

Introduction of alien species may significantly affect soil ecosystems, through predation or disruption of components of native ecosystems (Winsor et al. 2004; \Álvarez-Presas et al. 2014; Justine et al. 2014). Land planarians have been reported as alien species in soils throughout the world and, among those, some species are considered to be successful invaders, e.g. Platydemus manokwari de Beauchamp, 1963, Arthurdendyus triangulatus (Dendy, 1894), Bipalium adventitium Hyman, 1943, Bipalium kewense Moseley, 1878 and Dolichoplana striata Moseley, 1877 (Winsor et al. 2004; \Álvarez-Presas et al. 2014; Justine et al. 2014, 2015). Soil moisture status seems to be an important element for their successful invasion (Fraser \& Boag 1998). In Europe at least 18 species of alien land planarians have been recorded since now and some of them are considered as invasive ones, e.g. P. manokwari (cf. Justine et al. 2014). Although the alien land planarian B. kewense has been reported to occur in many greenhouses in Italy (Bello et al. 1995), no data are available on its establishment and/or impact on natural environments. On 28th September 2014, 20 specimens (~1 individual/m2) of the land planarian Diversibipalium multilineatum (Makino \& Shirasawa, 1983) (Fig. 1), native to Japan, were collected under pots, branches and plastic materials in a private garden located in the center of Bologna (Emilia Romagna, Central Italy), near the urban park Giardini Margherita (44\°29\&$\#$39; N, 11\°21\&$\#$39; E; WGS84). Thirty plant species (both indigenous and alien), mainly cultivated as bonsai (e.g. Lagerstroemia indica L., Juniperus procumbens (Siebold ex Endl.) Miquel), were present in this shady, wet garden (25 m2). Between March 2014 and June 2015, 70 more specimens of D. multilineatum were collected at the same site, mainly at dusk and dawn after rain. Reproduction by fission and regeneration processes were observed in several of those specimens, which were kept for some time in captivity. A specimen of D. multilineatum was also collected in a garden in L\éguevin (Haute-Garonne, France), which will be described in a forthcoming paper by Justine et al. (in prep.) (see also Kawakatsu et al. 2014). Specimens without a genital pore were initially ascribed to D. multilineatum on the basis of their external appearance: the dorsal surface was brownish yellow and presented five longitudinal stripes at the head plate and the neck, showing the typical appearance of the species. The middorsal stripe was widened at its anterior end, on the head plate, and at the pharynx level. The ventral pattern of the animals at the pharyngeal region was also characteristic, with the middorsal stripe widened at this level. The Italian Diversibipalium specimens used for the molecular analysis were fixed and preserved in absolute ethanol. Fragments of the mitochondrial gene COI and 28S ribosomal RNA nuclear gene (GenBank Acc. Numbers KU245358 and KU245357, respectively) were obtained using the procedure and COI primers described in \Álvarez-Presas et al. (2008) and Sol\à et al. (2013). The French specimen\&$\#$39;s COI (Specimen MNHN JL177, GenBank Acc. Number KT922162) was obtained as described in Justine et al. (2015). 28S sequences of 14 Bipaliinae specimens and four Microplana species (outgroup) retrieved from GenBank were included in the phylogenetic analyses (Fig. 2). Sequence alignment was obtained by using the online software MAFFT version 7 (Katoh \& Standley 2013), while ambiguously aligned positions were removed using the program Gblocks (Talavera \& Castresana 2007) with default settings, excepting the minimum number of sequences for a flank position at the minimum value (set at 10) and with half of the allowed gap positions. The final alignment had a length of 1589 bp. We used two phylogenetic inference approaches: maximum likelihood (ML), using the RaxML 8.2.3 software (Stamatakis 2014), and Bayesian inferences (BI), using MrBayes 3.2.4 (Ronquist et al. 2012). The evolutionary model used, GTR+I+G, was estimated to be the best with the software jModeltest 2.1.7 (Darriba et al. 2012; Guindon \& Gascuel 2003), using the Akaike Information Criterion (AIC). MrBayes analyses were performed for 10-milion generation with sampling parameters every 103 and a 25\% default burn-in value for the final trees. Convergence of the two runs (average standard deviation of split frequencies \<\< 0.01) and likelihood stationarity were checked. The maximum likelihood analyses were performed under 1000 bootstrap pseudoreplicates. The phylogenetic results show a close and highly supported relationship of the Italian Diversibipalium specimens with those from Japan and South Korea that have been identified as D. multilineatum (Fig. 2). Diversibipalium multilineatum is the sister-group of B. nobile Kawakatsu \& Makino, 1982, but with low support. The COI sequences of the French (MNHN JL177) and the Italian Diversibipalium specimens were compared in Geneious v. 8.0.5 (http://www.geneious.com, Kearse et al. 2012) and were found to be identical. These results indicate that the species introduced in both countries is the same, and most probably concerns the species D. multilineatum. The pathways of introduction of D. multilineatum are currently unknown, although a relationship between the horticultural trade and the introduction of alien land planarians is well known (\Álvarez-Presas et al. 2014 and references therein). Here we report the first occurrence of individuals of D. multilineatum outside Asia. The GenBank sequence of D. multilineatum from South Korea is not yet supported by a published description of the specimen, while it is debatable whether South Korea should be considered part of the natural range of D. multilineatum, which only seems to include Japan. In the present paper, we consider the South Korean animal to be an introduced specimen. Soil moisture status, temperature, and food availability are considered to be the main factors determining the presence of terrestrial planarians (Boag et al. 1998); the microclimatic conditions of the Italian garden were similar to plant nurseries and greenhouses, while an abundance of food was available, such as isopods [Porcellionides pruinosus (Brandt, 1833)], oligochaetes [Dendrobaena attemsi (Michaelsen, 1902) and several juveniles of Lumbricus spp.] and gastropods [Cernuella cisalpina (Rossmassler, 1837), Cornu aspersum (O.F. M\üller 1774), Deroceras reticulatum (O.F. M\üller, 1774), Discus rotundatus (O.F. M\üller, 1774), Limacus flavus (Linnaeus, 1758), Milax nigricans (Philippi, 1836), Papillifera papillaris (Linnaeus, 1758), Pomatias elegans (O.F. M\üller, 1774)]. Moreover, winter 2014 reached the highest temperatures and rainfall of the last two decades (source: CNR-ISAC, Bologna), thus favouring establishment and spread of D. multilineatum. The potential environmental impacts of some invasive flatworms are well documented (\Álvarez-Presas et al. 2014; Justine et al. 2014) and, even if these effects have not yet been assessed for D. multilineatum, the adoption of precautionary measures and of early intervention is here strongly recommended (Genovesi \& Shine 2004). Finally, knowledge of the introduction pathway(s), together with the analysis of prey preference and possible impact on the invertebrate fauna, will be essential to halt or at least to limit the spread of this introduced land flatworm.

}, keywords = {Animals, Biological Evolution, Body Size, Europe, phylogeny, planarians}, issn = {1175-5334}, doi = {10.11646/zootaxa.4067.5.4}, author = {Mazza, Giuseppe and Menchetti, Mattia and Sluys, Ronald and Sol{\`a}, Eduard and Riutort, Marta and Tricarico, Elena and Justine, Jean-Lou and Cavigioli, Luca and Mori, Emiliano} } @article {1681, title = {Towards a comprehensive, integrative analysis of the diversity of European microplaninid land flatworms (Platyhelminthes, Tricladida, Microplaninae), with the description of two peculiar new species}, journal = {Systematics and BiodiversitySystematics and Biodiversity}, volume = {14}, year = {2016}, note = {doi: 10.1080/14772000.2015.1103323}, month = {2016/01/02}, pages = {9 - 31}, publisher = {Taylor \& Francis}, abstract = {

The present study forms a first and major step towards a comprehensive morphological and molecular analysis of the species diversity of European microplaninid land planarians by presenting a molecular phylogenetic tree on the basis of alignments of the mitochondrial Cox1 gene from 158 specimens as well as a concatenated phylogeny (Cox1 and 18S genes) on the basis of 41 sequences for nine Microplana species included in this study. Genetic distances between and within known and new species were calculated. Combined morphological and molecular results facilitated an integrative delimitation of new species as well as the diagnosis of new populations of already known species. An integrative account is provided of two new and aberrant species from the Iberian Peninsula and southern France. Through the molecular approach a Confirmed Candidate Species was detected among the newly sampled populations. Further we document samples from new localities for five already known species as well as new sites for the Confirmed Candidate Species. The new data considerably expand the European range of several already known species included in the present study. The results of an in-depth study of the taxonomic literature, as well as original material, are documented since this was required for appropriate identification of the new materials. This part of the study resulted in a re-evaluation of the taxonomic status of several nominal species and in the following taxonomic conclusions: Rhynchodemus pyrenaicus is a species of Microplana and not Rhynchodemus; Rhynchodemus attemsi, R. peneckei, and R. henrici should be transferred to the genus Microplana; Microplana attemsi and M. peneckei are not junior synonyms of M. henrici; Microplana styriaca is not a synonym of M. terrestris; Rhynchodemus howesi is not a junior synonym of M. pyrenaica and belongs to the genus Rhynchodemus and not to Microplana; Microplana richardi is not a synonym of M. terrestris. The current diagnosis of the genus Microplana should be amended such that it refers to the situation that the species generally have two small eyes but occasionally may have multiple eyes.http://zoobank.org/urn:lsid:zoobank.org:pub:22B437AA-9D41-4DA0-AE3C-BD8F636CB96DThe present study forms a first and major step towards a comprehensive morphological and molecular analysis of the species diversity of European microplaninid land planarians by presenting a molecular phylogenetic tree on the basis of alignments of the mitochondrial Cox1 gene from 158 specimens as well as a concatenated phylogeny (Cox1 and 18S genes) on the basis of 41 sequences for nine Microplana species included in this study. Genetic distances between and within known and new species were calculated. Combined morphological and molecular results facilitated an integrative delimitation of new species as well as the diagnosis of new populations of already known species. An integrative account is provided of two new and aberrant species from the Iberian Peninsula and southern France. Through the molecular approach a Confirmed Candidate Species was detected among the newly sampled populations. Further we document samples from new localities for five already known species as well as new sites for the Confirmed Candidate Species. The new data considerably expand the European range of several already known species included in the present study. The results of an in-depth study of the taxonomic literature, as well as original material, are documented since this was required for appropriate identification of the new materials. This part of the study resulted in a re-evaluation of the taxonomic status of several nominal species and in the following taxonomic conclusions: Rhynchodemus pyrenaicus is a species of Microplana and not Rhynchodemus; Rhynchodemus attemsi, R. peneckei, and R. henrici should be transferred to the genus Microplana; Microplana attemsi and M. peneckei are not junior synonyms of M. henrici; Microplana styriaca is not a synonym of M. terrestris; Rhynchodemus howesi is not a junior synonym of M. pyrenaica and belongs to the genus Rhynchodemus and not to Microplana; Microplana richardi is not a synonym of M. terrestris. The current diagnosis of the genus Microplana should be amended such that it refers to the situation that the species generally have two small eyes but occasionally may have multiple eyes.http://zoobank.org/urn:lsid:zoobank.org:pub:22B437AA-9D41-4DA0-AE3C-BD8F636CB96D

}, isbn = {1477-2000}, url = {http://dx.doi.org/10.1080/14772000.2015.1103323}, author = {Sluys, Ronald and Mateos, Eduardo and Riutort, Marta and {\'A}lvarez-Presas, Marta} } @article {ZOJ:ZOJ12358, title = {The true identity of Obama (Platyhelminthes: Geoplanidae) flatworm spreading across Europe}, journal = {Zoological Journal of the Linnean Society}, volume = {177}, number = {1}, year = {2016}, pages = {5{\textendash}28}, abstract = {

Since 2008 there have been many records in Europe (British Isles, Spain, France, Italy) of a large terrestrial planarian morphologically very similar to the Brazilian species Obama marmorata. Sequences of mitochondrial (Cox1) and nuclear (18S, 28S, ITS-1 and EF) genes from European specimens and some from Brazil indicate that they belong to a species different from that of other specimens also collected in Brazil. Moreover, the phylogenetic results show that they are not sister-species. Histological sections of both Brazilian and European specimens reveal subtle morphological differences between the two species. Obama marmorata is confined to Brazil, and the second, herein described new species, Obama nungara sp. nov., is found in Brazil and Europe. These cryptic species may be syntopic in areas in Brazil. The new species occurs in human-modified environments both in Brazil and in Europe. We also conclude that the specimens from Spain and Argentina identified previously as Obama marmorata belong to the new species.

}, keywords = {BPP, Continenticola, Cox I gene, Geoplana, GMYC, histology, morphology, phylogeny, planarians}, issn = {1096-3642}, doi = {10.1111/zoj.12358}, url = {http://dx.doi.org/10.1111/zoj.12358}, author = {Carbayo, Fernando and {\'A}lvarez-Presas, Marta and Jones, Hugh D. and Riutort, Marta} } @article {1540, title = {Focus on the details: morphological evidence supports new cryptic land flatworm (Platyhelminthes) species revealed with molecules}, journal = {Organisms Diversity \& Evolution}, volume = {15}, year = {2015}, pages = {379-403}, author = {{\'A}lvarez-Presas, Marta and Amaral, Silvana and Carbayo, Fernando and Leal-Zanchet, Ana Maria and Riutort, Marta} } @article {1493, title = {Evolutionary Analysis of Mitogenomes from Parasitic and Free-Living Flatworms}, journal = {PLoS ONE}, volume = {10}, year = {2015}, month = {2015}, pages = {e0120081}, publisher = {Public Library of Science}, address = {San Francisco, CA USA}, abstract = {

Mitochondrial genomes (mitogenomes) are useful and relatively accessible sources of molecular data to explore and understand the evolutionary history and relationships of eukaryotic organisms across diverse taxonomic levels. The availability of complete mitogenomes from Platyhelminthes is limited; of the 40 or so published most are from parasitic flatworms (Neodermata). Here, we present the mitogenomes of two free-living flatworms (Tricladida): the complete genome of the freshwater species Crenobia alpina (Planariidae) and a nearly complete genome of the land planarian Obama sp. (Geoplanidae). Moreover, we have reanotated the published mitogenome of the species Dugesia japonica (Dugesiidae). This contribution almost doubles the total number of mtDNAs published for Tricladida, a species-rich group including model organisms and economically important invasive species. We took the opportunity to conduct comparative mitogenomic analyses between available free-living and selected parasitic flatworms in order to gain insights into the putative effect of life cycle on nucleotide composition through mutation and natural selection. Unexpectedly, we did not find any molecular hallmark of a selective relaxation in mitogenomes of parasitic flatworms; on the contrary, three out of the four studied free-living triclad mitogenomes exhibit higher A+T content and selective relaxation levels. Additionally, we provide new and valuable molecular data to develop markers for future phylogenetic studies on planariids and geoplanids.

}, isbn = {1932-6203}, url = {http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4368550/}, author = {Sol{\`a}, Eduard and {\'A}lvarez-Presas, Marta and Fr{\'\i}as-L{\'o}pez, Cristina and Littlewood, D Timothy J and Rozas, Julio and Riutort, Marta}, editor = {Escriva, Hector} } @article {1492, title = {The taxonomic status of Dugesia biblica from Israel and Turkey (Platyhelminthes, Tricladida, Dugesiidae)}, journal = {ZooKeys}, year = {2015}, month = {2015}, pages = {1 - 12}, publisher = {Pensoft Publishers}, abstract = {

The taxonomic status of Dugesia biblica (Platyhelminthes, Tricladida, Dugesiidae) from Israel and Turkey is problematic due to its morphological similarity with Dugesia sicula since these nominal species present overlapping characters. In this study we analyzed histological preparations of specimens of these two nominal species and also compared mitochondrial COI gene sequences from Israeli populations to the already known haplotype composition of Dugesia sicula. We concluded that these animals belong to the same species and therefore we consider Dugesia biblica to be a junior synonym of Dugesia sicula. This implies that the distribution range of Dugesia sicula is even wider than previously thought, and that the species is present all around the Mediterranean Basin and on many of its islands.

}, isbn = {1313-29891313-2970}, url = {http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4467178/}, author = {Sol{\`a}, Eduard and Sluys, Ronald and Segev, Ori and Blaustein, Leon and Riutort, Marta} }