Metazoans of redoxcline sediments in Mediterranean deep-sea hypersaline anoxic basins.

BACKGROUND: The deep-sea hypersaline anoxic basins (DHABs) of the Mediterranean (water depth ~3500 m) are some of the most extreme oceanic habitats known. Brines of DHABs are nearly saturated with salt, leading many to suspect they are uninhabitable for eukaryotes. While diverse bacterial and protistan communities are reported from some DHAB haloclines and brines, loriciferans are the only metazoan reported to inhabit the anoxic DHAB brines. Our goal was to further investigate metazoan communities in DHAB haloclines and brines. RESULTS: We report observations from sediments of three DHAB (Urania, Discovery, L'Atalante) haloclines, comparing these to observations from sediments underlying normoxic waters of typical Mediterranean salinity. Due to technical difficulties, sampling of the brines was not possible. Morphotype analysis indicates nematodes are the most abundant taxon; crustaceans, loriciferans and bryozoans were also noted. Among nematodes, Daptonema was the most abundant genus; three morphotypes were noted with a degree of endemicity. The majority of rRNA sequences were from planktonic taxa, suggesting that at least some individual metazoans were preserved and inactive. Nematode abundance data, in some cases determined from direct counts of sediments incubated in situ with CellTracker(TM) Green, was patchy but generally indicates the highest abundances in either normoxic control samples or in upper halocline samples; nematodes were absent or very rare in lower halocline samples. Ultrastructural analysis indicates the nematodes in L'Atalante normoxic control sediments were fit, while specimens from L'Atalante upper halocline were healthy or had only recently died and those from the lower halocline had no identifiable organelles. Loriciferans, which were only rarely encountered, were found in both normoxic control samples as well as in Discovery and L'Atalante haloclines. It is not clear how a metazoan taxon could remain viable under this wide range of conditions. CONCLUSIONS: We document a community of living nematodes in normoxic, normal saline deep-sea Mediterranean sediments and in the upper halocline portions of the DHABs. Occurrences of nematodes in mid-halocline and lower halocline samples did not provide compelling evidence of a living community in those zones. The possibility of a viable metazoan community in brines of DHABs is not supported by our data at this time.

Molecular phylogenetic position and description of a new genus and species of freshwater Chaetonotidae (Gastrotricha: Chaetonotida: Paucitubulatina), and the annotation of its mitochondrial genome.

Chaetonotidae is the most diversified family of the entire phylum Gastrotricha; it comprises ~430 species distributed across 16 genera. The current classification, established mainly on morphological traits, has been challenged in recent years by phylogenetic studies, indicating that the cuticular ornamentations used to discriminate among species may be misleading when used to identify groupings, which has been the practice until now. Therefore, a consensus is developing toward implementing novel approaches to better define species identity and affiliation at a higher taxonomic ranking. Using an integrative morphological and molecular approach, including annotation of the mitogenome, we report on some freshwater gastrotrichs characterised by a mixture of two types of cuticular scales diagnostic of the genera Aspidiophorus and Heterolepidoderma . Our specimens' overall anatomical characteristics find no correspondence in the taxa of these two genera, calling for their affiliation to a new species. Phylogenetic analyses based on the sequence of the ribosomal RNA genes of 96 taxa consistently found the new species unrelated to Aspidiophorus or Heterolepidoderma but allied with Chaetonotus aff. subtilis, as a subset of a larger clade, including mostly planktonic species. Morphological uniqueness and position along the non-monophyletic Chaetonotidae branch advocate erecting a new genus to accommodate the current specimens; consequently, the name Litigonotus ghinii gen. nov., sp. nov. is proposed. The complete mitochondrial genome of the new taxon resulted in a single circular molecule 14,384 bp long, including 13 protein-coding genes, 17 tRNA genes and 2 rRNAs genes, showing a perfect synteny and collinearity with the only other gastrotrich mitogenome available, a possible hint of a high level of conservation in the mitochondria of Chaetonotidae. ZooBank: urn:lsid:zoobank.org:pub:9803F659-306F-4EC3-A73B-8C704069F24A.

An annotated checklist of the chaetonotidan Gastrotricha from India.

India has a long history of research on freshwater and marine Gastrotricha. In more than 110 years of study on Order Chaetonotida, two families consisting of 11 genera and 39 species have been described. Thirty of these species are taxa originally described from other continents, while only nine species (7 freshwater, 2 marine) are only known from India. The large percentage (77%) of so-called cosmopolitan species in India has contributed to the phenomenon known as the meiofauna paradox. However, a careful review of the pertaining literature provides a different biogeographical picture of the chaetonotidan fauna of India. Herein we show that the high incidence of European and North American species reported from India is mainly due to a mixture of misidentification and species lumping. In fact, for only 12 species there are enough data that would make the Indian specimens morphological similar to taxa previously reported from Europe and/or North America. However, without the appropriate molecular sequence data for comparison, there is no way to rule out the possibility of cryptic speciation.We conclude that further sampling throughout India and the use of more powerful microscopical techniques (e.g., DIC optics) and molecular sequencing will reveal more species and improve the quality of re-descriptions of those (9 spp.) that so far appear to be endemic to the subcontinent. Here we recommend six species to be excluded from the fauna of India while another 11 species (non endemic to India) should be considered dubitatively present in the Indian fauna.

The curious and neglected soft-bodied meiofauna: Rouphozoa (Gastrotricha and Platyhelminthes).

Gastrotricha and Platyhelminthes form a clade called Rouphozoa. Representatives of both taxa are main components of meiofaunal communities, but their role in the trophic ecology of marine and freshwater communities is not sufficiently studied. Traditional collection methods for meiofauna are optimized for Ecdysozoa, and include the use of fixatives or flotation techniques that are unsuitable for the preservation and identification of soft-bodied meiofauna. As a result, rouphozoans are usually underestimated in conventional biodiversity surveys and ecological studies. Here, we give an updated outline of their diversity and taxonomy, with some phylogenetic considerations. We describe successfully tested techniques for their recovery and study, and emphasize current knowledge on the ecology, distribution and dispersal of freshwater gastrotrichs and microturbellarians. We also discuss the opportunities and pitfalls of (meta)barcoding studies as a means of overcoming the taxonomic impediment. Finally, we discuss the importance of rouphozoans in aquatic ecosystems and provide future research directions to fill in crucial gaps in the biology of these organisms needed for understanding their basic role in the ecology of benthos and their place in the trophic networks linking micro-, meio- and macrofauna of freshwater ecosystems.

Human access impacts biodiversity of microscopic animals in sandy beaches.

Whereas most work to understand impacts of humans on biodiversity on coastal areas has focused on large, conspicuous organisms, we highlight effects of tourist access on the diversity of microscopic marine animals (meiofauna). We used a DNA metabarcoding approach with an iterative and phylogeny-based approach for the taxonomic assignment of meiofauna and relate diversity patterns to the numbers of tourists accessing sandy beaches on an otherwise un-impacted island National Park. Tourist frequentation, independently of differences in sediment granulometry, beach length, and other potential confounding factors, affected meiofaunal diversity in the shallow "swash" zone right at the mean water mark; the impacts declined with water depth (up to 2 m). The indicated negative effect on meiofauna may have a consequence on all the biota including the higher trophic levels. Thus, we claim that it is important to consider restricting access to beaches in touristic areas, in order to preserve biodiversity.

An annotated checklist of the macrodasyidan Gastrotricha from India.

We present a checklist of the macrodasyidan Gastrotricha fauna of India derived from original articles and comprehensive monographies regarding the Indian Fauna. Taxa and specimens described from India are discussed with special attention to their status, especially those species previously known from distant geographic areas. A total of 43 species have been recorded from 4 States and 2 Union territories. A careful taxonomic assessment indicated that many species are in need of re-description as the reported anatomical traits are not sufficient to grant a modern, robust identification. The situation is particularly poor for two species (Macrodasys indicus Govindankutty Nair, 1969 and Paradasys lineatus Rao, 1980), which consequently, are considered here to be species inquirendae; moreover, the Indian records of Tetranchyroderma hystrix and T. massiliense reported from Kerala are doubtful and consequently the two species should not be included among the fauna of the Indian region until proper evidence is found. It is necessary to unravel the true identities of many Indian species through a comparison to topotypical material. We recommend that future comparisons between related species and conspecifics from different populations to include molecular sequence data.

Integrated data analysis allows the establishment of a new, cosmopolitan genus of marine Macrodasyida (Gastrotricha).

Macrodasyida (phylum Gastrotricha) comprises 365 species distributed across 34 genera and 10 families. However, current classification is under revision due to the contradictory results of molecular and morphological cladistic analyses. Studies aimed at bridging the gaps took advantage of supplementary assessments of poorly known species and particularly from observations of new taxa showing original traits that could help to identify plesiomorphic character states in these anatomically diverse micrometazoa. We follow this path by describing three new interesting macrodasyidan species respectively from Italy, Brazil and Sweden. In many respects, the new species resemble most closely species of the genus Macrodasys; however, details of the external morphology, in combination with the different lay-out of the reproductive system and the tiny spermatozoa lacking a visible flagellum, suggest they belong to a new genus, possibly in the family Macrodasyidae. These hypotheses are supported by the phylogenetic relationships of 47 taxa inferred from analyses of the 18S rRNA gene, which found the new species clustering with Thaidasys tongiorgii in a subset of a larger clade containing Macrodasys. Accordingly, the establishment of the following taxa is proposed: Kryptodasys gen. nov., K. marcocurinii sp. nov., K. carlosrochai sp. nov. and K. ulfjondeliusi sp. nov.

A new species of Turbanellidae (Gastrotricha, Macrodasyida) from Jamaica, with a key to species of Paraturbanella.

The study falls within the framework of a wider research programme aimed at investigating the gastrotrich diversity of the Tropical North-Western Atlantic (TNWA). A new macrodasyidan gastrotrich is described from fine-medium sand collected at Duncans Bay, Jamaica. The description is based on observations carried out on living specimens using differential interference contrast microscopy. Paraturbanella xaymacanasp. n., the third gastrotrich taxon reported from Jamaica, is a mid-sized species, up to 564 µm long, with a feeble peribuccal swelling. The most obvious autapomorphic traits pertain to the testes and the male pore, both of which are located approximately at mid body, rather than at- or near the pharyngo-intestinal junction as occur in the other species of the genus. Additional differences with congeners are discussed and a key to the Paraturbanella species is provided, in the hope it will be useful to both gastrotrich experts and marine ecologists who discover these microscopic metazoans during their research.

Erratum: Author Correction: Biodiversity estimates and ecological interpretations of meiofaunal communities are biased by the taxonomic approach.

[This corrects the article DOI: 10.1038/s42003-018-0119-2.].

Biodiversity estimates and ecological interpretations of meiofaunal communities are biased by the taxonomic approach.

Accurate assessments of biodiversity are crucial to advising ecosystem-monitoring programs and understanding ecosystem function. Nevertheless, a standard operating procedure to assess biodiversity accurately and consistently has not been established. This is especially true for meiofauna, a diverse community (>20 phyla) of small benthic invertebrates that have fundamental ecological roles. Recent studies show that metabarcoding is a cost-effective and time-effective method to estimate meiofauna biodiversity, in contrast to morphological-based taxonomy. Here, we compare biodiversity assessments of a diverse meiofaunal community derived by applying multiple taxonomic methods based on comparative morphology, molecular phylogenetic analysis, DNA barcoding of individual specimens, and metabarcoding of environmental DNA. We show that biodiversity estimates are strongly biased across taxonomic methods and phyla. Such biases affect understanding of community structures and ecological interpretations. This study supports the urgency of improving aspects of environmental high-throughput sequencing and the value of taxonomists in correctly understanding biodiversity estimates.

Two new marine Gastrotricha from the Indian Ocean coast of South Africa.

The study is part of a larger research programme aimed at shedding light on the gastrotrich communities of the subtropical east coast province of KwaZulu-Natal, South Africa. In previous papers, faunistic and preliminary taxonomic data on marine and freshwater gastrotrichs found in the iSimangaliso Wetland Park, were reported. Here two new interesting marine macrodasyidan species in the families Dactylopodolidae and Thaumastodermatidae are described based on observations carried out on living specimens and using differential interference contrast microscopy. The two novel species are named in honor of two great South African icons recently deceased: Nadine Gordimer and Nelson Mandela. Dactylopodola nadine sp. n. is the third species in the genus to bear red eye-spots; it can easily be distinguished from the closely-related red-eyed D. baltica and D. roscovita by its smaller size (Total length = 230 µm vs 275 µm vs 450 µm, respectively) and the lower number of adhesive tubes of the anterior, lateral and posterior series (on each side: 3, 4 and 4 vs 5, 6 and 8 vs 2, 9 and 12-15). Pseudostomella mandela sp. n. is a fairly large species (up to 481 µm in length), with a cuticular covering made up of tetrancres and relatively long caudal pedicles (up to 44 µm in length ). The most evident autoapomorphic trait of the new species is the presence of 7 pairs of 'cirrata' tubes, two emerging in a lateral position along the pharyngeal region and five from the dorsolateral sides of the trunk. Additional relevant taxonomic characters include: 4 tubes of the anterior series, 11 tubes of the ventrolateral series and 3 tubes of the posterior series per side, 5 papillae on the dorsal margin and 6 papillae on the ventral margin of the oral palps. The high number of putative new species discovered among the South African gastrotrich fauna during our relatively short survey, highlights the relevance of this region with regard to the diversity of this group and stresses once again the importance of investigating new geographic areas in order to improve our understanding of global gastrotrich biodiversity and species richness. 

An Integrated Morphological and Molecular Approach to the Description and Systematisation of a Novel Genus and Species of Macrodasyida (Gastrotricha).

BACKGROUND: Gastrotricha systematics is in a state of flux mainly due to the conflicts between cladistic studies base on molecular markers and the classical systematisation based on morphological traits. In sandy samples from Thailand, we found numerous macrodasyidan gastrotrichs belonging to an undescribed species of difficult taxonomic affiliation. The abundance and original nature of the specimens prompted us to undertake a deep survey of both morphological and molecular traits aiming at a reliable systematisation of the new taxon. METHODOLOGY/PRINCIPAL FINDINGS: Using several microscopical techniques we investigated the external and internal anatomy, including the muscular and nervous systems of the new species. Additional specimens were used to obtain the 18S rRNA gene sequence; molecular data was analysed cladistically in conjunction with data from additional species belonging to the near complete Macrodasyida taxonomic spectrum. Specimens are vermiform, up to 806 µm in total length, and show a well-defined head equipped with peculiar leaf-like sensorial organs and a single-lobed posterior end. The adhesive apparatus includes anterior, ventrolateral, dorsal and posterior tubes. Pharynx is about 1/4 of the total length and shows pores at its posterior 3/4. Adult specimens exhibit maturing eggs and a bulky, muscular caudal organ, but do not show sperm nor the frontal organ. Musculature and nervous system organisation resemble the usual macrodasyidan plan; however, the somatic circular muscles of the intestinal region surround all other muscular components and a third FMRFamide-IR commissure ventral to the pharyngo-intestinal junction appear to be an autoapomorphic traits of the new species. CONCLUSIONS/SIGNIFICANCE: While the anatomical characteristics of the Asian specimens appear so unique to grant the establishment of a new taxon, for which the name Thaidasys tongiorgii gen. et sp. nov. is proposed, the result of phylogenetic analyses based on the 18S rRNA gene unites the new genus with the family Macrodasyidae.

Fauna Europaea: Gastrotricha.

Fauna Europaea provides a public web-service with an index of scientific names (including important synonyms) of all living European land and freshwater animals, their geographical distribution at country level (up to the Urals, excluding the Caucasus region), and some additional information. The Fauna Europaea project covers about 230,000 taxonomic names, including 130,000 accepted species and 14,000 accepted subspecies, which is much more than the originally projected number of 100,000 species. This represents a huge effort by more than 400 contributing specialists throughout Europe and is a unique (standard) reference suitable for many users in science, government, industry, nature conservation and education. Gastrotricha are a meiobenthic phylum composed of 813 species known so far (2 orders, 17 families) of free-living microinvertebrates commonly present and actively moving on and into sediments of aquatic ecosystems, 339 of which live in fresh and brackish waters. The Fauna Europaea database includes 214 species of Chaetonotida (4 families) plus a single species of Macrodasyida incertae sedis. This paper deals with the 224 European freshwater species known so far, 9 of which, all of Chaetonotida, have been described subsequently and will be included in the next database version. Basic information on their biology and ecology are summarized, and a list of selected, main references is given. As a general conclusion the gastrotrich fauna from Europe is the best known compared with that of other continents, but shows some important gaps of knowledge in Eastern and Southern regions.

A transcriptomic-phylogenomic analysis of the evolutionary relationships of flatworms.

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.

A new non-naked species of Ptychostomella (Gastrotricha) from Brazil.

A new species of marine Gastrotricha from Brazil is described and discussed. Ptychostomella lamelliphora sp. n. is one of the several new taxa that were found during an extensive survey of the gastrotrich fauna carried out in 2002 and 2003 along the coastline of the State of São Paulo. The new species is unique in that it possesses cuticular ornamentations in the form of plate-like structures (scales) along the lateral borders of the body and two massive clusters of densely packed adhesive tubes on the ventral surface, near the ano-genital opening. Both these features appear to be adaptations to challenge the high energy waters that characterize the species' microhabitat: the coarse sublittoral sand in the channel between the mainland and the largest island in the State, Ihlabela. Additionally, a key to the described Ptychostomella species of the world is provided.

Neogosseidae (Gastrotricha, Chaetonotida) from the iSimangaliso Wetland Park, KwaZulu-Natal, South Africa.

Among the mostly benthic gastrotrichs, the Neogosseidae (Gastrotricha, Chaetonotida) are particularly interesting from an evolutionary point of view in virtue of their planktonic lifestyle; yet, they are poorly known and uncertainties concerning morphological traits hamper accurate in-group systematics. During a recent survey of meiofauna in the iSimangaliso Wetland Park, South Africa, two species of Neogosseidae were found in a freshwater pond near Charter's Creek on the Western Shores of Lake St Lucia. Based on morphological traits, one species has been identified as Neogossea acanthocolla, originally described from Brazil, while the other, affiliated to the genus Kijanebalola, is proposed as new to science. Using a combination of differential interference contrast and scanning electron microscopy, fine anatomical details were observed and are here discussed in a larger taxonomic framework, especially regarding Kijanebalola devestiva sp. n. Results have also provided reasons for a revision of the diagnostic traits of Kijanebalola, Neogossea and the whole Family Neogosseidae. Besides expanding awareness about the biodiversity hosted by South Africa's first UNESCO World Heritage Site, our study will be beneficial to future phylogenetic studies of the Gastrotricha based on morphology, by allowing the selection and/or a more precise character coding of traits of phylogenetic relevance.

A new marine gastrotrich from the State of São Paulo (Brazil), with a key to species of Pseudostomella (Gastrotricha, Thaumastodermatidae).

In previous papers, faunistic and preliminary taxonomic data on the gastrotrich communities along the coastline of the Brazilian states of São Paulo and Rio de Janeiro were reported; among the over 40 records, the occurrence of several species new to science was highlighted. One of such new taxa is described here based on observation carried out on living and SEM prepared specimens. Pseudostomella dolichopodasp. n. (Gastrotricha: Thaumastodermatidae) is the only species in the genus that attains 420 µm in total length, is covered by pentancres and possesses, among others, caudal pedicles up to 45 µm in length. Additional differences with co-generic taxa characterized by a pentancrous covering are discussed. Furthermore, a key to the described Pseudostomella species of the world based on easily discernible traits, visible in both living and formalin-fixed specimens, is provided.

Gastrotricha: a marine sister for a freshwater puzzle.

BACKGROUND: Within an evolutionary framework of Gastrotricha Marinellina flagellata and Redudasys fornerise bear special interest, as they are the only Macrodasyida that inhabit freshwater ecosystems. Notwithstanding, these rare animals are poorly known; found only once (Austria and Brazil), they are currently systematised as incertae sedis. Here we report on the rediscovery of Redudasys fornerise, provide an account on morphological novelties and present a hypothesis on its phylogenetic relationship based on molecular data. METHODOLOGY/PRINCIPAL FINDINGS: Specimens were surveyed using DIC microscopy and SEM, and used to obtain the 18 S rRNA gene sequence; molecular data was analyzed cladistically in conjunction with data from 42 additional species belonging to the near complete Macrodasyida taxonomic spectrum. Morphological analysis, while providing new information on taxonomically relevant traits (adhesive tubes, protonephridia and sensorial bristles), failed to detect elements of the male system, thus stressing the parthenogenetic nature of the Brazilian species. Phylogenetic analysis, carried out with ML, MP and Bayesian approaches, yielded topologies with strong nodal support and highly congruent with each other. Among the supported groups is the previously undocumented clade showing the alliance between Redudasys fornerise and Dactylopodola agadasys; other strongly sustained clades include the densely sampled families Thaumastodermatidae and Turbanellidae and most genera. CONCLUSIONS/SIGNIFICANCE: A reconsideration of the morphological traits of Dactylopodola agadasys in light of the new information on Redudasys fornerise makes the alliance between these two taxa very likely. As a result, we create Anandrodasys gen. nov. to contain members of the previously described D. agadasys and erect Redudasyidae fam. nov. to reflect this novel relationship between Anandrodasys and Redudasys. From an ecological perspective, the derived position of Redudasys, which is deeply nested within the Macrodasyida clade, unequivocally demonstrates that invasion of freshwater by gastrotrichs has taken place at least twice, in contrast with the single event hypothesis recently put forward.

Patterns of diversity in soft-bodied meiofauna: dispersal ability and body size matter.

BACKGROUND: Biogeographical and macroecological principles are derived from patterns of distribution in large organisms, whereas microscopic ones have often been considered uninteresting, because of their supposed wide distribution. Here, after reporting the results of an intensive faunistic survey of marine microscopic animals (meiofauna) in Northern Sardinia, we test for the effect of body size, dispersal ability, and habitat features on the patterns of distribution of several groups. METHODOLOGY/PRINCIPAL FINDINGS: As a dataset we use the results of a workshop held at La Maddalena (Sardinia, Italy) in September 2010, aimed at studying selected taxa of soft-bodied meiofauna (Acoela, Annelida, Gastrotricha, Nemertodermatida, Platyhelminthes and Rotifera), in conjunction with data on the same taxa obtained during a previous workshop hosted at Tjärnö (Western Sweden) in September 2007. Using linear mixed effects models and model averaging while accounting for sampling bias and potential pseudoreplication, we found evidence that: (1) meiofaunal groups with more restricted distribution are the ones with low dispersal potential; (2) meiofaunal groups with higher probability of finding new species for science are the ones with low dispersal potential; (3) the proportion of the global species pool of each meiofaunal group present in each area at the regional scale is negatively related to body size, and positively related to their occurrence in the endobenthic habitat. CONCLUSION/SIGNIFICANCE: Our macroecological analysis of meiofauna, in the framework of the ubiquity hypothesis for microscopic organisms, indicates that not only body size but mostly dispersal ability and also occurrence in the endobenthic habitat are important correlates of diversity for these understudied animals, with different importance at different spatial scales. Furthermore, since the Western Mediterranean is one of the best-studied areas in the world, the large number of undescribed species (37%) highlights that the census of marine meiofauna is still very far from being complete.

Phylogeny of Thaumastodermatidae (Gastrotricha: Macrodasyida) inferred from nuclear and mitochondrial sequence data.

BACKGROUND: Phylogenetic relationships within Gastrotricha are poorly known. Attempts to shed light on this subject using morphological traits have led to hypotheses lacking satisfactory statistical support; it seemed therefore that a different approach was needed. METHODOLOGY/PRINCIPAL FINDINGS: In this paper we attempt to elucidate the relationships within the taxonomically vast family Thaumastodermatidae (Macrodasyida) using molecular sequence data. The study includes representatives of all the extant genera of the family and for the first time uses a multi-gene approach to infer evolutionary liaisons within Gastrotricha. The final data set comprises sequences of three genes (18S, 28S rDNA and COI mtDNA) from 41 species, including 29 thaumastodermatids, 11 non-thaumastodermatid macrodasyidans and a single chaetonotidan. Molecular data was analyzed as a combined set of 3 genes and as individual genes, using Bayesian and maximum likelihood approaches. Two different outgroups were used: Xenotrichula intermedia (Chaetonotida) and members of the putative basal Dactylopodola (Macrodasyida). Thaumastodermatidae and all other sampled macrodasyidan families were found monophyletic except for Cephalodasyidae. Within Thaumastodermatidae Diplodasyinae and Thaumastodermatinae are monophyletic and so are most genera. Oregodasys turns out to be the most basal group within Thaumastodermatinae in analyses of the concatenated data set as well as in analyses of the nuclear genes. Thaumastoderma appears as the sister taxon to the remaining species. Surprisingly, Tetranchyroderma is non-monophyletic in our analyses as one group of species clusters with Ptychostomella while another appears as the sister group of Pseudostomella. CONCLUSIONS/SIGNIFICANCE: Results in general agree with the current classification; however, a revision of the more derived thaumastodermatid taxa seems necessary. We also found that the ostensible COI sequences from several species do not conform to the general invertebrate or any other published mitochondrial genetic code; they may be mitochondrially derived nuclear genes (numts), or one or more modifications of the mitochondrial genetic code within Gastrotricha.