Volume X-Ray Micro-Computed Tomography Analysis of the Early Cephalized Central Nervous System in a Marine Flatworm, Stylochoplana pusilla.

Platyhelminthes are a phylum of simple bilaterian invertebrates with prototypic body systems. Compared with non-bilaterians such as cnidarians, the bilaterians are likely to exhibit integrated free-moving behaviors, which require a concentrated nervous system "brain" rather than the distributed nervous system of radiatans. Marine flatworms have an early cephalized 'central' nervous system compared not only with non-bilaterians but also with parasitic flatworms or freshwater planarians. In this study, we used the marine flatworm Stylochoplana pusilla as an excellent model organism in Platyhelminthes because of the early cephalized central nervous system. Here, we investigated the three-dimensional structures of the flatworm central nervous system by the use of X-ray micro-computed tomography (micro-CT) in a synchrotron radiation facility. We found that the obtained tomographic images were sufficient to discriminate some characteristic structures of the nervous system, including nerve cords around the cephalic ganglion, mushroom body-like structures, and putative optic nerves forming an optic commissure-like structure. Through the micro-CT imaging, we could obtain undistorted serial section images, permitting us to visualize precise spatial relationships of neuronal subpopulations and nerve tracts. 3-D micro-CT is very effective in the volume analysis of the nervous system at the cellular level; the methodology is straightforward and could be applied to many other non-model organisms.

Two new Neotetraonchus species (Dactylogyridea, Dactylogyridae) parasitising the Peruvian sea catfish Galeichthys peruvianus (Siluriformes, Ariidae), including molecular data.

As part of a parasitological survey, several specimens of two new monopisthocotylean species, Neotetraonchus celsomanueli sp. nov. and N.peruvianus sp. nov. (Dactylogyridea, Dactylogyridae), were collected from the gill filaments of the Peruvian sea catfish Galeichthys peruvianus (Siluriformes, Ariidae) off Puerto Pizarro, Tumbes region, Peru. Neotetraonchus celsomanueli sp. nov. is characterised by an MCO with a T-shaped distal end and an accessory piece that is ribbed and expanded proximally with a worm-shaped termination. Neotetraonchus peruvianus sp. nov. is typified by its MCO, which has a sledgehammer-shaped distal end and an accessory piece with a claw-shaped distal end. Additionally, N.peruvianus sp. nov. is characterised by its jellyfish-shaped onchium. A partial 28S rDNA sequence was obtained from N.celsomanueli sp. nov., and a phylogenetic analysis was conducted. This analysis revealed the phylogenetic position of Neotetraonchus celsomanueli sp. nov. within a clade comprising monopisthocotylean parasites of diadromous and marine ariid catfishes, including Hamatopeduncularia spp., Chauhanellus spp., Thysanotohaptor Kritsky, Shameem, Kumari & Krishnaveni, , and Neocalceostomoides spinivaginalis Lim, 1995. This finding brings the number of known Neotetraonchus species to seven and represents the first described Neotetraonchus species infecting marine catfishes from Peru.

Species of Dactylogyridae (Monogenoidea, Polyonchoinea) from the gills of haemulid and lutjanid fishes (Perciformes) from the Pacific coast of Mexico.

Examination of the gill lamellae of the Panama grunt Rhencus panamensis (Steindachner) (Haemulidae), golden snapper Lutjanus inermis (Peters), and yellow snapper Lutjanus argentiventris (Peters) (Lutjanidae) (Perciformes) from the coast of the Guerrero State (eastern Tropical Pacific) of Mexico revealed 5 species of Monogenoidea: Euryhaliotrema disparum n. sp. on R. panamensis; Haliotrematoides uagroi n. sp. on L. inermis; and E. anecorhizion Kritsky & Mendoza-Franco, 2012, E. fastigatum (Zhukov, 1976) Kritsky & Boeger, 2002, and E. paracanthi (Zhukov, 1976) Kritsky & Boeger, 2002 on L. argentiventris. Specimens found on R. panamensis were assigned within Euryhaliotrema as a new species possessing the atypical morphology of the male copulatory organ (i.e., a coiled tube with clockwise rings). Haliotrematoides uagroi n. sp. differs from Haliotrematoides striatohamus (Zhukov, 1981) Mendoza-Franco, Reyes-Lizama & Gonzalez-Solis, 2009 from Haemulon spp. (Haemulidae) from the Caribbean Sea (Mexico) in having inner blades on the distal shafts of the ventral and dorsal anchors. The present paper represents the first finding of a species of Euryhaliotrema (E. disparum n. sp.) on a species of Rhencus and the second species on a haemulid host, and H. uagroi n. sp. as the first monogenoidean species described on L. inermis. Euryhaliotrema anecorhizion, E. fastigatum, and E. paracanthi on L. argentiventris in the Pacific coast of Mexico represent new geographical records.

Explosive networking: The role of adaptive host radiations and ecological opportunity in a species-rich host-parasite assembly.

Many species-rich ecological communities emerge from adaptive radiation events. Yet the effects of adaptive radiation on community assembly remain poorly understood. Here, we explore the well-documented radiations of African cichlid fishes and their interactions with the flatworm gill parasites Cichlidogyrus spp., including 10,529 reported infections and 477 different host-parasite combinations collected through a survey of peer-reviewed literature. We assess how evolutionary, ecological, and morphological parameters determine host-parasite meta-communities affected by adaptive radiation events through network metrics, host repertoire measures, and network link prediction. The hosts' evolutionary history mostly determined host repertoires of the parasites. Ecological and evolutionary parameters predicted host-parasite interactions. Generally, ecological opportunity and fitting have shaped cichlid-Cichlidogyrus meta-communities suggesting an invasive potential for hosts used in aquaculture. Meta-communities affected by adaptive radiations are increasingly specialised with higher environmental stability. These trends should be verified across other systems to infer generalities in the evolution of species-rich host-parasite networks.

Two New Brackish-Water Species of Macrostomum (Platyhelminthes: Macrostomorpha) from China and Their Phylogenetic Positions.

In this paper, two new brackish-water species of the macrostomid turbellarian genus Macrostomum, Macrostomum pseudosinense sp. nov. and Macrostomum taurinum sp. nov., collected from coastal water at Shenzhen, Guangdong Province, China, are described based on morphological, histological, and molecular phylogenetic analyses. Macrostomum pseudosinense sp. nov. differs from similar species within the genus in the length of the stylet (152 ± 15.0 µm), diameter of stylet opening (20 ± 4.0 µm proximally; 7 ± 0.5 µm distally), two bends of the stylet, and the non-spiral end of the stylet. Macrostomum taurinum sp. nov. differs from its congeners in the length of the stylet (81 ± 7.4 µm), the stylet bending position and angle (50% and 60°), diameter of stylet proximal opening (15 ± 3.0 µm), sperm with bristles and brush, and the smooth-walled ovaries. Phylogenetic analyses inferred from nuclear 18S and 28S rRNA genes support the establishments of these two new species. In addition, reciprocal mating behavior of M. pseudosinense sp. nov. was observed and documented.

RNA-Seq of three free-living flatworm species suggests rapid evolution of reproduction-related genes.

BACKGROUND: The genus Macrostomum consists of small free-living flatworms and contains Macrostomum lignano, which has been used in investigations of ageing, stem cell biology, bioadhesion, karyology, and sexual selection in hermaphrodites. Two types of mating behaviour occur within this genus. Some species, including M. lignano, mate via reciprocal copulation, where, in a single mating, both partners insert their male copulatory organ into the female storage organ and simultaneously donate and receive sperm. Other species mate via hypodermic insemination, where worms use a needle-like copulatory organ to inject sperm into the tissue of the partner. These contrasting mating behaviours are associated with striking differences in sperm and copulatory organ morphology. Here we expand the genomic resources within the genus to representatives of both behaviour types and investigate whether genes vary in their rate of evolution depending on their putative function. RESULTS: We present de novo assembled transcriptomes of three Macrostomum species, namely M. hystrix, a close relative of M. lignano that mates via hypodermic insemination, M. spirale, a more distantly related species that mates via reciprocal copulation, and finally M. pusillum, which represents a clade that is only distantly related to the other three species and also mates via hypodermic insemination. We infer 23,764 sets of homologous genes and annotate them using experimental evidence from M. lignano. Across the genus, we identify 521 gene families with conserved patterns of differential expression between juvenile vs. adult worms and 185 gene families with a putative expression in the testes that are restricted to the two reciprocally mating species. Further, we show that homologs of putative reproduction-related genes have a higher protein divergence across the four species than genes lacking such annotations and that they are more difficult to identify across the four species, indicating that these genes evolve more rapidly, while genes involved in neoblast function are more conserved. CONCLUSIONS: This study improves the genus Macrostomum as a model system, by providing resources for the targeted investigation of gene function in a broad range of species. And we, for the first time, show that reproduction-related genes evolve at an accelerated rate in flatworms.

Geographical Distribution and Genetic Diversity of Praesagittifera naikaiensis (Acoelomorpha) in the Seto Inland Sea, Japan.

Acoel flatworms are simple bilaterians that lack digestive lumens and coelomic cavities. Although they are a significant taxon for evaluating the evolution of metazoans, suitable species for biological experiments are not available in Japan. We recently focused on Praesagittifera naikaiensis, which inhabits the sandy shores of intertidal zones in the Seto Inland Sea in Japan, as a candidate for a representative acoel species to be used in experiments. However, reports on its distribution range remain limited. Here, we surveyed the habitats of P. naikaiensis on 108 beaches along the Seto Inland Sea. Praesagittifera naikaiensis is reported here from 37 sites (six previously known and 31 newly discovered sites) spread over a wide area of the Seto Inland Sea, from Awaji Island in Hyogo Prefecture to Fukuoka Prefecture (364 km direct distance). Based on the mitochondrial cytochrome oxidase subunit I (COI) gene haplotypes, we evaluated the genetic diversity of 145 individuals collected from 33 sites. Out of 42 COI haplotypes, 13 haplotypes were shared by multiple individuals. The most frequent haplotype was observed in 67 individuals collected from 31 sites. Eight other haplotypes were detected at geographically distant locations (maximum of 299 km direct distance). Multiple haplotypes were found at 32 sites. These results demonstrate that sufficient genetic flow exists among P. naikaiensis populations throughout the Seto Inland Sea. Molecular phylogenetic analysis of the COI haplotypes of P. naikaiensis revealed that all specimens were grouped into one clade. The genetic homogeneity of the animals in this area favors their use as an experimental animal.

On Temnocephala axenos Monticelli, 1898 (Platyhelminthes, Temnocephalida): taxonomic status and designation of a neotype.

Temnocephala axenos Monticelli, 1898 was described based on specimens from an unidentified host collected in Blumenau, Santa Catarina, Brazil. Information about type locality was imprecise and the host was later identified as Aegla laevis (Latreille, 1818). However, it is known that A. laevis is not present on the eastern side of the Andes. Also, only histological preparations from one specimen studied by Monticelli are currently available in the Museum für Naturkunde Berlin, but it showed none of the taxonomic characters needed for the characterization of the species. Although the updated description of the species based on Uruguayan specimens, neither the author nor the several previous studies about the species showed a search for the type material, a resolution for the misidentification of the type host or the imprecise type locality due to the subsequent geographical division of the municipality cited in the description. The Uruguayan specimens were not even geographically close to the type locality and a neotype was not designed to validate the species' taxonomic status again. Specimens from Santa Catarina and Paraná States, Brazil, were studied, as well as restudied Argentinean specimens. The new data were compared with the update description of the species. The historical background and the discussion about geographical origins and hosts of the species, as well as a designation of a neotype, allow comparative material of the type locality and type host to exist, eliminating doubts about the identification of T. axenos.

Rapid divergence, molecular evolution, and morphological diversification of coastal host-parasite systems from southern Brazil.

This study assessed the role of historical processes on the geographic isolation, molecular evolution, and morphological diversification of host-parasite populations from the southern Brazilian coast. Adult specimens of Scleromystax barbatus and Scleromystax macropterus were collected from the sub-basin of the Nhundiaquara River and the sub-basin of the Paranaguá Bay, state of Paraná, Brazil. Four species of Gyrodactylus were recovered from the body surface of both host species. Morphometric analysis of Gyrodactylus spp. and Scleromystax spp. indicated that subpopulations of parasites and hosts could be distinguished from different sub-basins and locations, but the degree of morphological differentiation seems to be little related to geographic distance between subpopulations. Phylogenetic relationships based on DNA sequences of Gyrodactylus spp. and Scleromystax spp. allowed distinguishing lineages of parasites and hosts from different sub-basins. However, the level of genetic structuring of parasites was higher in comparison to host species. Evidence of positive selection in mtDNA sequences is likely associated with local adaptation of lineages of parasites and hosts. A historical demographic analysis revealed that populations of Gyrodactylus and Scleromystax have expanded in the last 250 000 years. The genetic variation of parasites and hosts is consistent with population-specific selection, population expansions, and recent evolutionary co-divergence.

Rhinoxenus (Dactylogyridae) parasitizing piranhas (Serrasalmidae) at its southernmost limit of distribution (Paraná River, Argentina), with the description of two new species.

Three piranha species, Serrasalmus maculatus, S. marginatus and Pygocentrus nattereri, living sympatrically in the lower Paraná River (Argentina) were examined searching for nasal monogeneans to know its diversity and distribution. Four species of monogeneans belonging to Rhinoxenus were found parasitizing the nasal cavities. Two new species are described, and new morphological data of 2 previously described species is provided. Rhinoxenus argentinensis n. sp. is characterized by having the male copulatory organ (MCO) as a coiled tube with a reel shaped-base; an elongated accessory piece articulated to base of MCO; a sinistral vagina with a sclerotized vestibule, and a sclerotized cap of the ventral anchor laterally modified forming a triangular expansion. Rhinoxenus paranaensis n. sp. is characterized by having a MCO as a coiled tube with a reel-shaped base; an accessory piece with an elongate proximal portion, a dilated distal portion with digitiform projections articulated to base of MCO; a sinistral vagina with 5-6 cuticular spine-shaped processes, and a sclerotized cap of the ventral anchor modified laterally forming a bilobate expansion. Additionally, multivariate discriminant analyses showed significant morphometric differences in the ventral anchors among Rhinoxenus species parasitizing 'piranhas '.

The first ultrastructural observations of the egg-forming complex of Calicotyle affinis, an endoparasitic monocotylid monogenean which lacks a uterus.

This ultrastructural study of the female reproductive system of Calicotyle affinis, a monogenean without a uterus, demonstrates, for the first time in a monogenean, the presence of accessory cells located in the peripheral region of the ovary and a single large cell whose surface is penetrated by deep invaginations which restrict the passage of oocytes through the lumen of the distal extremity of the ovary. The cytoarchitecture of the epithelial lining of the two vaginae is a syncytium formed by an anucleate epithelial lining with sunken epithelial perykaria. The wall of the seminal receptacle is an enlarged prolongation of the vaginal epithelial lining. Fertilization occurs in the fertilization chamber and fertilized oocytes retain cortical granules within their cytoplasm, a characteristic known to occur in free-living flatworms. Our study also highlights concentrations of two distinct groups of Mehlis' gland cell ducts on either side of the proximal end of the ootype distinguished by ultrastructural characteristics of their secretory granules. The epithelial wall of the ootype is formed by a single layer of regular, columnar, glandular epithelial cells; these cells are closely adjacent, conjoined towards their apical region by septate junctions and produce rounded, electron-dense granules which are discharged into the ootype lumen via a merocrine or holocrine mechanism. Released granules concentrate around the eggshell and form an additional fibrous coat. The morphological diversity observed in the female reproductive system of the Monogenea is commented on it relation to clarifying patterns in monogenean evolution and for understanding the phylogeny of the Neodermata.

Supplemental diagnosis of Gyrodactylus fairporti Van Cleave, 1921 (Monogenea) from 1 month old black bullhead (Ameiurus melas) in riverside pools in Wisconsin.

Samples of Gyrodactylus fairporti Van Cleave, 1921 from young-of-the-year black bullhead (Ameiurus melas) stranded in riverside pools of the Black River (La Crosse County,) WI, USA, are used to supplement the species diagnosis, including new details on the marginal hook sickles, the male copulatory organ (MCO), and 18S rRNA gene sequence data. The anchors of G. fairporti are relatively long and thin, 58.2 ± 1.2 µm in length; roots 15.5 ± 1.0 µm; shaft 38.1 ± 1.5 µm; point 31.3 ± 1.5 µm. The ventral bar is 19.4 ± 0.4 µm wide and 5.7 ± 0.9 µm long, with small anterolateral processes, 2.0 ± 0.6 µm long, and an almost rectangular posterior shield 15.5 ± 1.1 µm in length. The marginal hooks are 29.2 ± 1.0 µm long, with the handle 23.9 ± 1.2 µm in length. These measurements are similar to those reported from Iowa and Alabama, with the additional observation of the anchor point bending outwardly halfway along its length. The marginal hook sickle blade leaves the base angled ventrally away from the longitudinal axis; the sickle point is short; the toe has a rounded shelf and the heel is small, thin, and rounded. The MCO has eight small spines, two large ones laterally and the others of various lengths, with two of the smallest spines being slightly out-of-line compared with all the others. The taxonomy of G. fairporti is compared to Gyrodactylus ictaluri Rogers, 1967 and G. nebulosus Kritsky & Mizelle, 1967, the other two species known from captive and wild ictalurids endemic to North America. The three species all have a relatively compact ventral bar with short anterolateral processes, a short almost rectangular ventral bar membrane, an MCO with up to eight small spines of varying length, and a hook sickle angled ventrally. Diagnostically, the species are readily identified by the total length and shape of the anchors. G. fairporti bears the longest (53-65 µm) and most slender anchors of the trio, G. nebulosus intermediate (49-51 µm) and G. ictaluri with the shortest and stoutest (40-45 µm) of these species. A BLAST search of a partial (413 bp) 18S rRNA gene showed the highest similarity with Gyrodactylus sp. reported from Ameiurus nebulosus (Siluriformes) in Ontario.

Dactylogyrids (Platyhelminthes, Monogenoidea) from the gills of Hoplias malabaricus (Characiformes: Erythrinidae) from coastal rivers of the Oriental Amazon Basin: species of Urocleidoides and Constrictoanchoratus n. gen.

Five species of Urocleidoides (one new) and two new species of Constrictoanchoratus n. gen. are described in this study. All were collected from the gills of Hoplias malabaricus (Characiformes: Erythrinidae) captured in six localities of coastal rivers of the north-eastern sector the State of Pará (Oriental Amazon): Urocleidoides brasiliensis Rosim, Mendoza-Franco & Luque, 2011; Urocleidoides bulbophallus n. sp.; Urocleidoides cuiabai Rosim, Mendoza-Franco & Luque, 2011; Urocleidoides eremitus Kritsky, Thatcher & Boeger, 1986; Urocleidoides malabaricusi Rosim, Mendoza-Franco & Luque, 2011; Constrictoanchoratus lemmyi n. gen. n. sp.; and Constrictoanchoratus ptilonophallus n. gen. n. sp. This is the first reported occurrence of the four previously described species of Urocleidoides parasitizing H. malabaricus from streams in the Oriental Amazon Basin. The analysis of voucher specimens of U. eremitus parasitizing the gills of H. malabaricus from the Upper Paraná River floodplain in the limits of States of Paraná and Mato Grosso do Sul, Brazil, indicates that these specimens are members of a new species of Urocleidoides, described here as Urocleidoides paranae n. sp. Constrictoanchoratus n. gen. is proposed for the species with a male copulatory organ sclerotized, coiled, clockwise; ventral anchor with elongate superficial root, inconspicuous deep root; dorsal anchor with inconspicuous roots, and a constriction at the intersection between the shaft and the point. The host-parasite diversity scenario and host specificity of the species of Constrictoanchoratus n. gen. and Urocleidoides from the gills of H. malabaricus are also discussed in this study.

A new species of Tereancistrum (Monogenea, Dactylogyridae) from the gills of three Leporinus species (Characiformes, Anostomidae) and a revised description of Tereancistrum parvus.

The present study describes Tereancistrum flabellum n. sp. (Dactylogyridae, Ancyrocephalinae) from the gills of the anostomid fishes Leporinus friderici, Leporinus amblyrhynchus and Leporinus elongatus from two freshwater ecosystems in the south east of Brazil. This new species is mainly characterized by the morphology of the copulatory complex (such as the MCO base formed by two fan-shaped structures, and accessory piece flattened, curved, rigid and channeled), a dorsal anchor with a well-developed superficial and inconspicuous deep root, and the shape of the accessory anchor sclerite with small spathulate termination. Tereancistrum flabellum n. sp. is the first record of a dactylogyrid from L. amblyrhynchus. The description of Tereancistrum parvus is also emended to correct and complement previous descriptions and the species is reported for the first time in Schizodon nasutus.

New evidence of a lateral transfer of monogenean parasite between distant fish hosts in Lake Ossa, South Cameroon: the case of Quadriacanthus euzeti n. sp.

Species of the monogenean genus Quadriacanthus mainly infect fish belonging to the Siluriformes, especially the genera Clarias, Heterobranchus or Bagrus, and their host specificity is strict (oioxenous) or narrow (stenoxenous). An examination of the gills of 19 Papyrocranus afer from Lake Ossa, South Cameroon, revealed for the first time the presence of a species of Quadriacanthus from a fish host belonging to the Notopteridae. The morphology and the size of sclerotized parts of haptor and the male and female copulatory complexes suggest that this monogenean is a new species named Quadriacanthus euzeti n. sp. The fish genus Papyrocranus differs taxonomically from the usual fish hosts of Quadriacanthus and hence the presence of a species belonging to this genus on the gills of this host suggests the occurrence of a lateral transfer of Quadriacanthus from species belonging to Clarias or Bagrus which live sympatrically with P. afer in Lake Ossa.

Morphology and molecular analysis of Mizelleus indicus Jain () and M. longicirrus (Tripathi, ) Venkatanarasaiah & Kulkarni 1981 (Monogenea, Dactylogyridae) from the freshwater shark Wallago attu in the Ganga River, India.

Species of the genus Mizelleus Jain (1957) have always been controversial regarding identification and validity. Members of this group of species differ from each other in the morphology of their hard parts, which can be misleading and subject to differing interpretation among scientists. Therefore, the main objective of present study was to identify Mizelleus worms by morphological methods and molecular analysis on the basis of 18S ribosomal DNA to clarify their phylogenetic status. In this study, specimens were isolated from the gill filaments of Wallago attu (Siluriformes) and studied morphologically. In accordance with morphological characters, the specimens were found to be Mizelleus indicus and Mizelleus longicirrus. Partial sequences of nuclear 18S rDNA of these two species were amplified. The results confirm the phylogenetic relationships and taxonomic validation of M. indicus and M. longicirrus in India.

Elucidating the phylogenetic position of Gnathostomulida and first mitochondrial genomes of Gnathostomulida, Gastrotricha and Polycladida (Platyhelminthes).

Gnathostomulida is a taxon of small marine worms, which exclusively inhabit the interstitium. The evolution of Gnathostomulida has been discussed for decades. Originally regarded as primitive animals with affinities to flatworms, the phylogenetic position of Gnathostomulida has been debated. Given the lack of an anus a close relationship to Platyhelminthes has been maintained (i.e., Plathelminthomorpha hypothesis). Alternative hypotheses proposed Gnathostomulida as being close to Gastrotricha due to the presence of a monociliary epidermis (i.e., Monokonta/Neotrichozoa hypothesis) or to Syndermata based on the complicated jaw apparatus (i.e., Gnathifera hypothesis). Molecular analyses using only few genes were inconclusive. Recent phylogenomic studies brought some progress by placing Gnathostomulida as sister to Syndermata, but support for this relationship was low and depended on the analytical strategy. Herein we present the first data of complete or nearly complete mitochondrial genomes for two gnathostomulids (Gnathostomula paradoxa &G. armata), one gastrotrich (Lepidodermella squamata) and one polyclad flatworm (Stylochoplana maculata) to address the uncertain phylogenetic affinity of Gnathostomulida. Our analyses found Gnathostomulida as sister to Syndermata (Gnathifera hypothesis). Thorough sensitivity analyses addressing taxon instability, branch length heterogeneity (also known as long branch attraction) and base composition heterogeneity showed that the position of Gnathostomulida is consistent across the different analyses and, hence, independent of potential misleading biases. Moreover, by ameliorating these different biases nodal support values could be increased to maximum values. Thus, our data support the hypothesis that the different jaw apparatuses of Syndermata and Gnathostomulida are indeed homologous structures as proposed by the Gnathifera hypothesis.

The first multi-gene phylogeny of the Macrostomorpha sheds light on the evolution of sexual and asexual reproduction in basal Platyhelminthes.

The Macrostomorpha-an early branching and species-rich clade of free-living flatworms-is attracting interest because it contains Macrostomum lignano, a versatile model organism increasingly used in evolutionary, developmental, and molecular biology. We elucidate the macrostomorphan molecular phylogeny inferred from both nuclear (18S and 28S rDNA) and mitochondrial (16S rDNA and COI) marker genes from 40 representatives. Although our phylogeny does not recover the Macrostomorpha as a statistically supported monophyletic grouping, it (i) confirms many taxa previously proposed based on morphological evidence, (ii) permits the first placement of many families and genera, and (iii) reveals a number of unexpected placements. Specifically, Myozona and Bradynectes are outside the three classic families (Macrostomidae, Microstomidae and Dolichomacrostomidae) and the asexually fissioning Myomacrostomum belongs to a new subfamily, the Myozonariinae nov. subfam. (Dolichomacrostomidae), rather than diverging early. While this represents the first evidence for asexuality among the Dolichomacrostomidae, we show that fissioning also occurs in another Myozonariinae, Myozonaria fissipara nov. sp. Together with the placement of the (also fissioning) Microstomidae, namely as the sister taxon of Dolichomacrostomidae, this suggests that fissioning is not basal within the Macrostomorpha, but rather restricted to the new taxon Dolichomicrostomida (Dolichomacrostomidae+Microstomidae). Furthermore, our phylogeny allows new insights into the evolution of the reproductive system, as ancestral state reconstructions reveal convergent evolution of gonads, and male and female genitalia. Finally, the convergent evolution of sperm storage organs in the female genitalia appears to be linked to the widespread occurrence of hypodermic insemination among the Macrostomorpha.

The nervous system of Xenacoelomorpha: a genomic perspective.

Xenacoelomorpha is, most probably, a monophyletic group that includes three clades: Acoela, Nemertodermatida and Xenoturbellida. The group still has contentious phylogenetic affinities; though most authors place it as the sister group of the remaining bilaterians, some would include it as a fourth phylum within the Deuterostomia. Over the past few years, our group, along with others, has undertaken a systematic study of the microscopic anatomy of these worms; our main aim is to understand the structure and development of the nervous system. This research plan has been aided by the use of molecular/developmental tools, the most important of which has been the sequencing of the complete genomes and transcriptomes of different members of the three clades. The data obtained has been used to analyse the evolutionary history of gene families and to study their expression patterns during development, in both space and time. A major focus of our research is the origin of 'cephalized' (centralized) nervous systems. How complex brains are assembled from simpler neuronal arrays has been a matter of intense debate for at least 100 years. We are now tackling this issue using Xenacoelomorpha models. These represent an ideal system for this work because the members of the three clades have nervous systems with different degrees of cephalization; from the relatively simple sub-epithelial net of Xenoturbella to the compact brain of acoels. How this process of 'progressive' cephalization is reflected in the genomes or transcriptomes of these three groups of animals is the subject of this paper.

A Flatworm from the Genus Waminoa (Acoela: Convolutidae) Associated with Bleached Corals in Western Australia.

A flatworm isolated from bleached colonies of the coral Coscinaraea marshae at Rottnest Island, Western Australia, is described using a combination of morphological and molecular systematics. This flatworm shares morphological features characteristic of the genus Waminoa (Acoelomorpha: Acoela), including the presence of two algal symbionts, but appears to have genital regions different from those of other described species of Waminoa. The design of new oligonucleotide primers enabled the amplification of partial 18S rDNA of the Rottnest Island acoel specimens, and phylogenetic analysis positioned them within Waminoa, confirming their placement in the genus. Furthermore, Waminoa specimens from Rottnest Island grouped into a sister clade to Waminoa brickneri, indicating that the morphological and genetic differences observed are most likely intraspecific and due to geographic variation. As such, we name these Rottnest Island specimens W. cf. brickneri, but highlight that key differences warrant further exploration before assignment to this species can be confirmed. This is the first acoel flatworm described from Western Australia and contributes to our understanding of the diversity and evolutionary relationship of the Acoela.