The interrelationships of the flatworms (phylum Platyhelminthes) are poorly resolved despite decades of morphological and molecular phylogenetic studies. The earliest-branching clades (Catenulida, Macrostomorpha, and Polycladida) share spiral cleavage and entolecithal eggs with other lophotrochozoans. Lecithoepitheliata have primitive spiral cleavage but derived ectolecithal eggs. Other orders (Rhabdocoela, Proseriata, Tricladida and relatives, and Bothrioplanida) all have derived ectolecithal eggs but have uncertain affinities to one another. The orders of parasitic Neodermata emerge from an uncertain position from within these ectolecithal classes. To tackle these problems, we have sequenced transcriptomes from 18 flatworms and 5 other metazoan groups. The addition of published data produces an alignment of >107,000 amino acids with less than 28% missing data from 27 flatworm taxa in 11 orders covering all major clades. Our phylogenetic analyses show that Platyhelminthes consist of the two clades Catenulida and Rhabditophora. Within Rhabditophora, we show the earliest-emerging branch is Macrostomorpha, not Polycladida. We show Lecithoepitheliata are not members of Neoophora but are sister group of Polycladida, implying independent origins of the ectolecithal eggs found in Lecithoepitheliata and Neoophora. We resolve Rhabdocoela as the most basally branching euneoophoran taxon. Tricladida, Bothrioplanida, and Neodermata constitute a group that appears to have lost both spiral cleavage and centrosomes. We identify Bothrioplanida as the long-sought closest free-living sister group of the parasitic Neodermata. Among parasitic orders, we show that Cestoda are closer to Trematoda than to Monogenea, rejecting the concept of the Cercomeromorpha. Our results have important implications for understanding the evolution of this major phylum.

Two conflicting morphological approaches to polyclad systematics highlight the relevance of molecular data for resolving the interrelationships of Polycladida. In the present study, phylogenetic trees were reconstructed based on a short alignment of the 28S rDNA marker gene with 118 polyclad terminals (24 new) including 100 different polyclad species from 44 genera and 22 families, as well as on a combined dataset using 18S and 28S rDNA genes with 27 polyclad terminals (19 new) covering 26 different polyclad species. In both approaches, Theamatidae and Cestoplanidae were included, two families that have previously been shown to switch from Acotylea to Cotylea. Three different alignment methods were used, both with and without alignment curation by Gblocks, and all alignments were subjected to Bayesian inference and maximum likelihood tree calculations. Over all trees of the combined dataset, an extended majority-rule consensus tree had weak support for Theamatidae and Cestoplanidae as acotyleans, and also the cotylean generaBoninia,ChromyellaandPericelisappeared as acotyleans. With the most inclusive short 28S dataset, on the other hand, there is good support for the aforementioned taxa as cotyleans. Especially with the short 28S matrix, taxon sampling, outgroup selection, alignment method and curation, as well as model choice were all decisive for tree topology. Well-supported parts of the phylogeny over all trees include Pseudocerotoidea, Prosthiostomoidea, Stylochoidea, Leptoplanoidea and Cryptoceloidea, the latter three with new definitions. Unstable positions in the tree were found not only for Theamatidae, Cestoplanidae,Boninia,ChromyellaandPericelis, but also forAnonymus,ChromoplanaandCycloporus.

The systematics and distribution of the order Polycladida within the Macaronesian archipelagos are analysed. New species (Marcusia alba sp. nov., Prostheceraeus crisostomum sp. nov., Parviplana sodade sp. nov., Euplana claridade sp. nov., Stylochus salis sp. nov. and Distylochus fundae sp. nov.), new variety (Pseudoceros rawlinsonae var. galaxy), new records and records of shared species among different archipelagos are studied to compare the marine flatworm biodiversity of each island. The complex of archipelagos known as Macaronesia (including Madeira, Selvagens Islands, Canary Islands, Azores and Cape Verde) share a volcanic origin and European political influence. The five archipelagos are located along the eastern coast of the Atlantic Ocean and are subject to similar trade winds, streams (like the Gulf Stream) and cold currents. The term Macaronesia has suffered several changes throughout the years and it still is a topic of discussion in present times. The new delimitation of Macaronesia is mainly based on systematic studies on the invertebrate fauna of the islands. The resulting analyses shed new light on the differences and similarities among these archipelagos. In addition, molecular analyses employing 28S nuclear gene sequences are compared to verify relationships among anatomically similar species of marine polyclads.

A study of polyclad fauna of the Atlantic coast of the Iberian Peninsula was carried out from 2010 to 2013. The paper reports nine new records belonging to three Cotylean families: the family Euryleptidae Lang, 1884, Pseudocerotidae Lang, 1884 and the family Prosthiostomidae Lang, 1884, and describes one new species, Euryleptodes galikias sp. n.