Post-Embryonic Development and Genital-Complex Formation in Three Species of Polyclad Flatworms.

Without the establishment of effective culturing systems, little can be known about the late developmental stages of polyclad flatworms. Here, we report a laboratory culturing system for three polyclad species: Comoplana pusilla, Notocomplana koreana, and Pseudostylochus obscurus, and we describe changes in their morphology from hatching to reproductive maturity. These species hatch out as lobe-less larvae with four eyespots, but the number of eyespots increases in later development. Cross-like and triangularly shaped larvae are observed in N. koreana and P. obscurus, respectively. After settlement, a pale area appears on the body of juveniles and then develops into the copulatory complexes. All three species could be successfully reared on brine shrimp, but only C. pusilla and N. koreana achieved reproductive maturation in such a culturing system. In P. obscurus, switching the food to the gastropod Monodonta labio induced sexual maturation.

Nervous system and musculature of the parasitic turbellarian Notentera ivanovi (Plathelminthes, Fecampiida).

The article reports the first description of the architecture of the musculature and cholinergic nervous system of the parasitic turbellarian Notentera ivanovi from the White Sea using histochemistry and confocal scanning laser microscopy. It has been demonstrated that the body wall is composed of layers of circular and longitudinal muscles between which there are diagonal muscles described here for the first time. The nervous system is of the regular closely spaced orthogon type with pronounced radiality. In order to make it clear whether the orthogon of this type is a phylogenetic characteristic of this group of worms or its shape is associated with the flat rounded body, the study of other representatives of the Fecampiida group is required.

Syndesmis aethopharynx (Umagillidae, Rhabdocoela, Platyhelminthes) from the sea urchin Paracentrotus lividus: First record from the Eastern Mediterranean, phylogenetic position and intraspecific morphological variation.

Specimens of Syndesmis aethopharynx Westervelt & Kozloff, 1990 (Umagillidae, Rhabdocoela, Platyhelminthes) were collected from the intestine of several specimens of the sea urchin Paracentrotus lividus (Lamarck, 1816) [2], Hansson, 2001 at the Greek coast. This represents the first report of a species of Syndesmis from Greece. Our study has revealed several previously-unreported morphological details and intraspecific variation, which are added to the species description. The position of S. aethopharynx within Umagillidae is confirmed for the first time through molecular data (based on nuclear 18S rDNA), using both Bayesian and maximum likelihood analyses.

Contribution to the knowledge of cotylean flatworms (Turbellaria, Polycladida) from Iranian coasts: Introducing a new species, with remarks on new records.

Very little information is available regarding marine free-living flatworms not only from Iran, but throughout the countries surrounding the Persian Gulf and the Gulf of Oman. The present study first introduces a new euryleptid species, and then reports four pseudocerotid polyclads which inhabit Iranian shallow rocky shores of the Persian Gulf and the Gulf of Oman. Maritigrella makranica sp. nov. is characterized dorsally by a medial cream or white reticulated appearance containing pale orange spots in a honeycomb pattern, a distinct orange submarginal band around the entire body margin and between the marginal tentacles, black spots scattered around mid-dorsal surface, becoming more sparse on raised median region and towards body margin, surrounded by a dark-grey halo around the body midline and orange-black halo towards margin. Three of the four pseudocerotids species belonging to the genera Pseudobiceros Faubel, 1984; Pseudoceros Lang, 1884; and Thysanozoon Grube, 1840, are new records for the studied areas, while the other has been reported in the Persian Gulf previously. Comments on Iranian species are provided and associations of flatworms with ascidians and sponges were observed. 

A new Imogine species (Turbellaria: Polycladida: Stylochidae) associated with rock oysters (Saccostrea cucullata) from the Persian Gulf, with a review of the genus.

A new species of acotylean polyclad, Imogine qeshmensis sp. nov. is described from Qeshm Island, Persian Gulf, Iran. I. qeshmensis is characterized by the number and arrangement of the tentacular, cerebral and frontal eyes, a body margin with opaque white bands and eyespots and a male genital complex with an highly muscular seminal vesicle equal in size to the prostatic vesicle.

How the worm got its pharynx: phylogeny, classification and Bayesian assessment of character evolution in Acoela.

Acoela are marine microscopic worms currently thought to be the sister taxon of all other bilaterians. Acoels have long been used as models in evolutionary scenarios, and generalized conclusions about acoel and bilaterian ancestral features are frequently drawn from studies of single acoel species. There is no extensive phylogenetic study of Acoela and the taxonomy of the 380 species is chaotic. Here we use two nuclear ribosomal genes and one mitochondrial gene in combination with 37 morphological characters in an analysis of 126 acoel terminals (about one-third of the described species) to estimate the phylogeny and character evolution of Acoela. We present an estimate of posterior probabilities for ancestral character states at 31 control nodes in the phylogeny. The overall reconstruction signal based on the shape of the posterior distribution of character states was computed for all morphological characters and control nodes to assess how well these were reconstructed. The body-wall musculature appears more clearly reconstructed than the reproductive organs. Posterior similarity to the root was calculated by averaging the divergence between the posterior distributions at the nodes and the root over all morphological characters. Diopisthoporidae is the sister group to all other acoels and has the highest posterior similarity to the root. Convolutidae, including several "model" acoels, is most divergent. Finally, we present a phylogenetic classification of Acoela down to the family level where six previous family level taxa are synonymized.

Morphology of the muscle articulation joint between the hooks of a flatworm (Kalyptorhynchia, Cheliplana sp.).

Schizorhynch kalyptorhynchs are meiofaunal turbellarian predators that possess an eversible proboscis that can be armed with two stout hooks. The hooks grasp and manipulate prey using a wide range of rotations and translations. These diverse motions are possible because the hook supports may function as a muscle articulation type joint-that is, a joint formed of muscle and connective tissue that connects, separates, and moves the microscopic hooks. We analyze the morphology of the flexible joint in a species of Cheliplana by using three types of microscopy: light, laser scanning confocal, and transmission electron. Radial myofilament bundles are present in the core of the hook supports, and lateral divaricator muscle fibers are located on their lateral surfaces. We develop a novel model for movements of the proboscis and describe the tensile function of the basement membrane that surrounds each hook support's medial glandular region. Contraction of divaricator muscle fibers antagonized by contraction of radial myofilaments causes the lateral bending of the hook supports and opening of the hook apparatus. Relaxation of the divaricator fibers and maximal contraction of the radial myofilaments, which put the medial basement membranes in tension, may cause medial bending in the hook supports and closing of the hook apparatus. During proboscis retraction, closure may also be aided by the compression of the hook apparatus as the proboscis is drawn through the rostral pore. The study provides new insights into the principles of support and movement in muscle articulations.

Bryoplana xerophila n. g. n. sp., a new limnoterrestrial microturbellarian (Platyhelminthes, Typhloplanidae, Protoplanellinae) from epilithic mosses, with notes on its ecology.

Bryoplana xerophila, a new genus and species of limnoterrestrial protoplanelline platyhelminth, was found in moss and soil covering a concrete wall in northern Alabama, USA. Bryoplana xerophila is the first taxon of limnoterrestrial Protoplanellinae recorded from North America and is one of the few rhabdocoels known from dry habitats. It is unique within Protoplanellinae in lacking rhabdites, having a pharynx rosulatus in the frontal half of the body, and lacking sclerotized parts in the male system. Notes on encystment, reproduction and feeding behavior are given. An updated identification key to all known genera of Protoplanellinae is presented.

Sex allocation predicts mating rate in a simultaneous hermaphrodite.

Sexual selection theory for separate-sexed animals predicts that the sexes differ in the benefit they can obtain from multiple mating. Conventional sex roles assume that the relationship between the number of mates and the fitness of an individual is steeper in males compared with females. Under these conditions, males are expected to be more eager to mate, whereas females are expected to be choosier. Here we hypothesize that the sex allocation, i.e. the reproductive investment devoted to the male versus female function, can be an important predictor of the mating strategy in simultaneous hermaphrodites. We argue that within-species variation in sex allocation can cause differences in the proportional fitness gain derived through each sex function. Individuals should therefore adjust their mating strategy in a way that is more beneficial to the sex function that is relatively more pronounced. To test this, we experimentally manipulated the sex allocation in a simultaneously hermaphroditic flatworm and investigated whether this affects the mating behaviour. The results demonstrate that individuals with a more male-biased sex allocation (i.e. relatively large testes and small ovaries) are more eager to mate compared with individuals with a more female-biased sex allocation (i.e. relatively small testes and large ovaries). We argue that this pattern is comparable to conventional gender roles in separate-sexed organisms.

Sex allocation and sexual conflict in simultaneously hermaphroditic animals.

Links between sex allocation (SA) and sexual conflict in simultaneous hermaphrodites have been evident since Charnov's landmark paper published 30 years ago. We discuss two links, namely the potential for sexual conflict over SA between sperm donor and recipient, and the importance of post-copulatory sexual selection and the resulting sexual conflict for the evolution of SA. We cover the little empirical and theoretical work exploring these links, and present an experimental test of one theoretical prediction. The link between SA and sexual conflict is an interesting field for future empirical and theoretical research.

[The phylogenetic role of ontogenies (recapitulation and morphogenesis)].

Different approaches to evolutionary interpretation of ontogenies are compared, with special emphasis on the evolutionary role of morphogenetic mechanisms (construction technologies) substantially affecting the structure of definitive forms: they largely determine the structural characteristics of organs, types of anatomical and histological systems, and specificity of symmetry of organisms and their parts. The role of cellular morphogenesis inherited from protozoic ancestors in the morphogenesis of multicellular organisms is demonstrated. Two main ways of improving morphogeneses are considered, based on epithelial morphogenesis and early determined few-celled primordial. On the one hand, the phylogenetic role of archallaxes and deviations is emphasized, these events often switching evolution to a fundamentally new direction. On the other hand, many characteristics of developmental stages are explainable by rationalization of morphogeneses and do not recapitulate ancestral forms, which should be taken into consideration in phylogenetic interpretation of embryogeneses; in particular, this applies to interpretation of axial relationships.

Determinants of mating and sperm-transfer success in a simultaneous hermaphrodite.

The number of mating partners an individual has within a population is a crucial parameter in sex allocation theory for simultaneous hermaphrodites because it is predicted to be one of the main parameters influencing sex allocation. However, little is known about the factors that determine the number of mates in simultaneous hermaphrodites. Furthermore, in order to understand the benefits obtained by resource allocation into the male function it is important to identify the factors that predict sperm-transfer success, i.e. the number of sperm a donor manages to store in a mate. In this study we experimentally tested how social group size (i.e. the number of all potential mates within a population) and density affect the number of mates and sperm-transfer success in the outcrossing hermaphroditic flatworm Macrostomum lignano. In addition, we assessed whether these parameters covary with morphological traits, such as body size, testis size and genital morphology. For this we used a method, which allows tracking sperm of a labelled donor in an unlabelled mate. We found considerable variation in the number of mates and sperm-transfer success between individuals. The number of mates increased with social group size, and was higher in worms with larger testes, but there was no effect of density. Similarly, sperm-transfer success was affected by social group size and testis size, but in addition this parameter was influenced by genital morphology. Our study demonstrates for the first time that the social context and the morphology of sperm donors are important predictors of the number of mates and sperm-transfer success in a simultaneous hermaphrodite. Based on these findings, we hypothesize that sex allocation influences the mating behaviour and outcome of sperm competition.

Anatomy and ultrastructure of the female reproductive system of Pleioplana atomata (Platyhelminthes: Polycladida).

The ultrastructure of the female reproductive system of the polyclad flatworm Pleioplana atomata is described. Numerous ovaries are scattered throughout the entire body but are mainly concentrated on the dorsal side. Within an ovary, a germinative zone with oogonia and prefolicular cells is located in the dorsal part of the ovary. The remaining part of the gonad is filled with previtellogenic and early vitellogenic oocytes enwrapped by follicular cells. During previtellogenesis, oocytes produce numerous eggshell globules, which are distributed into the cortical area of the cell in later stages. Eventually, these globules release their contents into the space between the eggshell cover and oolemma. Similar types of globules are also found in others flatworms, and may represent useful phylogenetic characters. Entolecital, vitellogenic oocytes pass to paired uteri, where vitellogenesis is completed. The remainder of the female reproductive system consists of paired thin uterine ducts that join a vagina. The distal part of the long, curved vagina forms a large Lang's vesicle, while the proximal part is connected to a female atrium leading to a female gonopore. We hypothesize that Lang's vesicle functions in the digestion of excess sperm received. Two kinds of different shell (cement) glands that release their secretion into the vagina are identified. Both are unicellular glands and each gland cell connects to the lumen of the vagina via an individual canal. Similar glands in other acotylean polyclads have been implicated in the formation of eggshell covers.

Structure and evolution of the pharynx simplex in acoel flatworms (Acoela).

The homology of pharynges within the mostly pharynx-less Acoela has been a matter of discussion for decades and even the basic question of whether a pharynx is a primitive trait within the Acoela and homologous to the pharynx of platyhelminth turbellarians is open. By using fluorescence staining of musculature, as well as conventional histological techniques and transmission electron microscopy, the present study sets focus on the mouth and pharynx (where present) of seven species of Acoela within Paratomellidae, Solenofilomorphidae, Hofsteniidae, Proporidae, and Convolutidae, as well as one species of Nemertodermatida and Catenulida, respectively. It is shown that among the investigated families of acoels there is a great variability in muscle systems associated with the mouth and pharynx and that pharynx histology and ultrastructural characters are widely diverse. There are no close similarities between the acoel pharynges and the catenulid pharynx but there is a general resemblance of the musculature associated with the mouth in the representatives of Paratomellidae and Nemertodermatida. On the basis of the profound differences in pharynx morphology, three major conclusions are drawn: 1) the pharynges as present in Recent acoels are not homologous to the pharynx simplex characteristic for Catenulida and Macrostomida within the Platyhelminthes; 2) the different muscular pharynx types of acoels are not homologous between higher taxa and thus a single acoel-type pharynx simplex cannot be defined; 3) the presence of a muscular pharynx most likely does not represent the ancestral state.

Redescription and designation of a neotype of Temnocephala talicei Dioni, 1967 (Platyhelminthes: Temnocephalida).

Temnocephala talicei was described from Uruguayan material found mainly on Aegla prado. Its original description was based principally on its distinctive penial stylet, and, therefore, many aspects of the species anatomy that are now important in the taxonomy of the genus were not contemplated; type specimens were never designated; and the material used to describe the species has been lost. The present work provides a redescription of the species based upon material collected from the type host and in the type locality. Given the possibility of confusion in its identification, and in order to preserve its taxonomic stability, a neotype of T. talicei is designated. The closest species to T. talicei is Temnocephala mertoni, from which it can be separated by the presence of conspicuous intestinal septa, a markedly asymmetrical sphincter in the vagina, markedly lobed testes, a straight penial stylet with a non-sinuous distal portion of the shaft, a characteristic large, 'mouthpiece'-shaped introvert without discrete thickenings, and approximately 10 crowns of well-developed spines of decreasing length placed throughout the whole extension of the introvert. A comparison of material from Uruguay, Argentina, and Paraguay shows that there is little variation in the morphometry of the species. Finally, a comparison of the post-tentacular syncytia of T. talicei and T. mertoni shows that this is not always a species-specific character.

First report of the introduction of an Australian temnocephalidan into the New World.

One of the preferred freshwater crayfish species in astaciculture is the large parastacid Cherax quadricarinatus, a native to Australia. This crayfish yields good production in warm water, and, therefore, it has been translocated from Australia to other countries. One such country is Uruguay, where the initial attempt in astaciculture is being conducted. In a Uruguayan crayfish farm, a large number of Diceratocephala boschmai (Platyhelminthes, Temnocephalida) were found on the body surface and branchial chamber of C. quadricarinatus. This is the first report of an introduced temnocephalidan in the New World, and of D. boschmai in a locality outside Australia/New Guinea. The original description of D. boschmai was based on a few characters that separated this species from the other Temnocephalida; therefore, a detailed description of the Uruguayan population of D. boschmai is also provided. The conditions in which D. boschmai is found in Uruguay are suitable for its development and reproduction. This raises concern about the effect that its release into the environment might have.

Acoel development indicates the independent evolution of the bilaterian mouth and anus.

Most bilaterian animals possess a through gut with a separate mouth and anus. It is commonly believed that during the transition from radial to bilateral symmetry, both openings evolved simultaneously by the lateral closure of a slit-like blastopore. Molecular phylogenies however, place the acoel flatworms, which have only one opening to their digestive system, as the sister group to all remaining Bilateria. To address how this single body opening is related to the mouth and anus of the protostomes and deuterostomes, we studied the expression of genes involved in bilaterian foregut and hindgut patterning during the development of the acoel Convolutriloba longifissura. Here we show that the genes brachyury and goosecoid are expressed in association with the acoel mouth, suggesting that this single opening is homologous to the mouth of other bilaterians. In addition, we find that the genes caudal, orthopedia and brachyury-which are expressed in various bilaterian hindguts-are expressed in a small region at the posterior end of the animal, separated from the anterior oral brachyury-expressing region by a dorsal domain of ectodermal bmp2/4 expression. These results contradict the hypothesis that the bilaterian mouth and anus evolved simultaneously from a common blastoporal opening, and suggest that a through gut might have evolved independently in different animal lineages.

Resource-dependent sex-allocation in a simultaneous hermaphrodite.

Most sex allocation theory is based on the relationship between the resource investment into male and female reproduction and the consequent fitness returns (often called fitness-gain curves). Here we investigate the effects of resource availability on the sex allocation of a simultaneously hermaphroditic animal, the free-living flatworm Macrostomum lignano. We kept the worms under different resource levels and determined the size of their testes and ovaries over a period of time. At higher resource levels, worms allocated relatively more into the female function, suggesting a saturating male fitness-gain curve for this species. A large part of the observed effect was due to a correlated increase in body size, showing size-dependent sex allocation in M. lignano. However, a significant part of the overall effect was independent of body size, and therefore likely due to the differences in resource availability. Moreover, in accordance with a saturating male fitness-gain curve, the worms developed the male gonads first. As the group size was kept constant, our results contrast with expectations from sex allocation models that deal with local mate competition alone, and with previous experiments that test these models.

A new Paravortex (Platyhelminthes, Dalyellioida) endoparasite of Mesodesma mactroides (Bivalvia, Mesodesmatidae) from Uruguay.

Many species of turbellarians (Platyhelminthes) are known to live associated with other organisms, especially invertebrates, as commensals or parasites. The family Graffillidae (Rhabdocoela) includes two genera that parasitize mollusks, Graffílla and Paravortex. Within the latter genus, six species were described as associated with mollusks. In other instances, unnamed Paravortex species were mentioned as parasites of other bivalves and of the body surface of fishes. In the present work, a new Paravortex species that was found in the intestine of Mesodesma mactroides from the Atlantic coast of Uruguay is described. In addition, a bibliographical revision of the known Paravortex species with their respective hosts, location, and distribution is made. Paravortex nicolli, described by Szidat for the Argentinean coast, is mentioned for the first time after the original description, and the authorship and date of description of Paravortex tapetis Noury-Sraïri 1989 are elucidated.

Morphology and ultrastructure of the pharynx in Solenofilomorphidae (Acoela).

The homology of pharynges within the mostly pharynx-less Acoela has been a matter of discussion for decades. Here, we analyze the pharynges of three members of the Solenofilomorphidae, Myopea sp. and two species of the genus Solenofilomorpha, by means of light and transmission electron microscopy. Special focus is placed on the ultrastructure of the pharyngeal musculature, epidermis surrounding the mouth, pharyngeal epithelium, and junction with the digestive parenchyma. The main goal of this study was to evaluate the usefulness of certain characters for broader comparisons within the Acoela. Among the three species, characters relating to position of the mouth, presence and elaboration of sphincter muscles, presence of pharyngeal glands, and ultrastructure of epitheliosomes proved to be variously species- and genus-specific. The arrangement of pharyngeal muscles and their connection with body wall musculature, ultrastructure of receptor cells, and morphology of a nonciliated glandular region in the posterior pharynx, in contrast, appear to be characteristic of the family Solenofilomorphidae and thus of predominant interest for comparisons with other acoel families.