Myzostomida: a link between trochozoans and flatworms?

Myzostomids are obligate symbiotic invertebrates associated with echinoderms with a fossil record that extends to the Ordovician period. Due to their long history as host-specific symbionts, myzostomids have acquired a unique anatomy that obscures their phylogenetic affinities to other metazoans: they are incompletely segmented, parenchymous, acoelomate organisms with chaetae and a trochophore larva. Today, they are most often classified within annelids either as an aberrant family of polychaetes or as a separate class. We inferred the phylogenetic position of the Myzostomida by analysing the DNA sequences of two slowly evolving nuclear genes: the small subunit ribosomal RNA and elongation factor-1alpha. All our analyses congruently indicated that myzostomids are not annelids but suggested instead that they are more closely related to flatworms than to any trochozoan taxon. These results, together with recent analyses of the myzostomidan ultrastructure, have significant implications for understanding the evolution of metazoan body plans, as major characters (segmentation, coeloms, chaetae and trochophore larvae) might have been independently lost or gained in different animal phyla.     

Mitochondrial genome and nuclear sequence data support myzostomida as part of the annelid radiation

The echinoderm symbionts Myzostomida are marine worms that show an enigmatic lophotrochozoan body plan. Historically, their phylogenetic origins were obscured due to disagreement about which morphological features are evolutionarily conserved, but now most morphological evidence points to annelid origins. In contrast, recent phylogenetic analyses using different molecular markers produced variable results regarding the position of myzostomids, but all suggested these worms are not derived annelids. To reexamine this issue, we analyzed data from nuclear genes (18S rDNA, 28S rDNA, Myosin II, and Elongation Factor-1alpha), and a nearly complete myzostomid mitochondrial genome. Here, we show that the molecular data are in agreement with the morphological evidence that myzostomids are part of the annelid radiation. This result is robustly supported by mitochondrial (gene order and sequence data) and nuclear data, as well as by recent ultrastructural investigations. Using Bayes factor comparison, alternative hypotheses are shown to lack support. Thus, myzostomids probably evolved from a segmented ancestor and gained a derived anatomy during their long evolutionary history as echinoderm symbionts.     

Evidence for cospeciation events in the host–symbiont system involving crinoids (Echinodermata) and their obligate associates,the myzostomids (Myzostomida,Annelida)

Although molecular-based phylogenetic studies of hosts and their associates are increasingly common in the literature, no study to date has examined the hypothesis of coevolutionary process between hosts and commensals in the marine environment. The present work investigates the phylogenetic relationships among 16 species of obligate symbiont marine worms (Myzostomida) and their echinoderm hosts (Crinoidea) in order to estimate the phylogenetic congruence existing between the two lineages. The combination of a high species diversity in myzostomids, their host specificity, their wide variety of lifestyles and body shapes, and millions years of association, raises many questions about the underlying mechanisms triggering their diversification. The phylogenetic relationships, inferred using a three-genes dataset (18S rDNA, 16S rDNA, and COI) and two-genes dataset (18S rDNA, and COI) for the myzostomids and crinoids, respectively, were congruent with the literature. The overall congruence between the two phylogenies was statistically significant according to topology-based, distance-based, and data-based approaches: a significant pattern of cophylogeny was found, though not perfect probably resulting from occasional host switches, duplications or extinction events. A minimum of 8 cospeciation events was estimated, which is significantly higher than it would have been expected due to chance alone.     

Locomotion and fine structure of parapodia in <Emphasis Type="Italic">Myzostoma cirriferum</Emphasis> (Myzostomida)

Most myzostomids are ectocommensals of crinoids on which they move freely. Their locomotion is ensured by five pairs of parapodia located laterally below their trunk. Each parapodium in Myzostoma cirriferum is a conical structure that includes a hook-like chaeta, replacement chaetae and an aciculum. Structure and ultrastructure of the myzostomid chaetae are similar to those of polychaetes: they are formed by a chaetoblast, which gives rise to microvilli where chaetal material is assembled on the outer surface. Myzostoma cirriferum walks on its host. It moves the anterior part, the posterior part or the lateral parts forwards but is able to rotate of 180° on itself. Its locomotion entirely depends on parapodial motions and not on trunk movements. Three pairs of muscles are involved in parapodial motions: parapodium flexor and parapodium extensor, aciculum protractor and aciculum retractor, and hook protractor with conjunctor. A functional model is proposed for explaining the global motion of a parapodium in M. cirriferum that may be extended to all ectocommensal myzostomids.     

Myoanatomy of Myzostoma cirriferum (Annelida,Myzostomida): Implications for the evolution of the myzostomid body plan

Studies of rare genomic marker systems suggest that Myzostomida are a subgroup of Annelida and phylogenomic analyses indicate an early divergence of this taxon within annelids. However, adult myzostomids show a highly specialized body plan, which lacks typical annelid features, such as external body annulation, coelomic cavities with metanephridia, and segmental ganglia of the nervous system. The putative loss of these features might be due to the parasitic/symbiotic lifestyle of myzostomids associated with echinoderms. In contrast, the larval anatomy and adult locomotory system resemble those of annelids. To clarify whether the myoanatomy of myzostomids reflects their relationship to annelids, we analyzed the distribution of f‐actin, a common component of muscle fibers, in specimens of Myzostoma cirriferum using phalloidin‐rhodamine labeling in conjunction with confocal laser‐scanning microscopy. Our data reveal that the musculature of the myzostomid body comprises an outer circular layer, an inner longitudinal layer, numerous dorsoventral muscles, and prominent muscles of the parapodial complex. These features correspond well with the common organization of the muscular system in Annelida. In contrast to other annelids, however, several elements of the muscular system in M. cirriferum, including the musculature of the body wall, and the parapodial flexor muscles, exhibit radial symmetry overlaying a bilateral body plan. These findings are in line with the annelid affinity of myzostomids and suggest that the apparent partial radial symmetry of M. cirriferum arose secondarily in this species. Based on our data, we provide a scenario on the rearrangements of muscle fibers that might have taken place in the lineage leading to this species. J. Morphol., 2013. © 2012 Wiley Periodicals, Inc.     

Molecular phylogenetic analyses indicate multiple independent emergences of parasitism in Myzostomida (Protostomia)

The fossil record indicates that Myzostomida, an enigmatic group of marine worms, traditionally considered as annelids, have exhibited a symbiotic relationship with echinoderms, especially crinoids, for nearly 350 million years. All known extant myzostomids are associated with echinoderms and infest their integument, gonads, celom, or digestive system. Using nuclear (18S rDNA) and mitochondrial (16S and COI) DNA sequence data from 37 myzostomid species representing nine genera, we report here the first molecular phylogeny of the Myzostomida and investigate the evolution of their various symbiotic associations. Our analyses indicate that the two orders Proboscidea and Pharyngidea do not constitute natural groupings. Character reconstruction analyses strongly suggest that (1) the ancestor of all extant myzostomids was an ectocommensal that first infested crinoids, and then asteroids and ophiuroids, and (2) parasitism in myzostomids emerged multiple times independently.     

PHYLOGENETIC ANALYSIS OF REPRODUCTIVE TRAITS OF MANIRAPTORAN THEROPODS AND ITS IMPLICATIONS FOR EGG PARATAXONOMY

Abstract:  A phylogenetic analysis of reproductive and oological (egg) traits of theropod taxa allows determination of the sequence in which these traits evolved in Theropoda. Our results indicate that several avian reproductive traits, such as adults sitting on eggs, asymmetrical eggs, unornamented eggshell surface, and complex eggshell ultrastructure, were already present in non-avian maniraptorans, and could have evolved in more basal theropods. In addition, non-avian maniraptorans laid two eggs at a time and orientated their eggs subvertically or subhorizontally in their nests, features not retained by neornithine birds. Based on our cladistic analysis it is also possible to infer the phylogenetic affinity of ootaxa of unknown parentage: Protoceratopsidovum was laid by a maniraptoran more derived than oviraptorids, and Parvoolithus probably belonged to a Cretaceous bird. Finally, our analysis reveals that many of the high-level categories of egg parataxonomy (morphotypes and basic types) are unnatural groupings (i.e. non-monophyletic). We recommend that these high-level categories be abandoned because oofamilies are sufficient to categorize egg taxa.     

Male reproductive system of the red brocade hermit crab Dardanus insignis (Diogenidae) and its relationship to other family members

The reproductive system of hermit crabs shows species-specific morphology, which can be used in phylogenetic analysis. Here, we describe the male reproductive system of the hermit crab Dardanus insignis, including morphological and biometric analyses of the spermatophore, the gonopore, and sperm ultrastructure. The morphological analyses were based on 15 selected specimens and carried out by means of light and electron microscopy. Our results indicate a reproductive system composed of lobular testes attached to a simple straight vas deferens connected to the exterior via ventral gonopores. The gonopores are ovoid, surrounded by dense serrulate setae, and covered by a membranous operculum. The spermatophores exhibit a tripartite structure, with an elongate ovoid ampulla, a long narrow stalk, and a proximal foot. The spermatozoal ultrastructure shows three main regions: an ovoid-oblong acrosomal vesicle, a nucleus, and cytoplasm with three armlike extensions. Some of these characteristics can also be found in other species of Diogenidae within the genus Dardanus and in members of Coenobitidae, a closely related family. The available information on spermatophore and spermatozoal structure may indicate a closer similarity between the genus Dardanus and the Coenobitidae, compared with other members of Diogenidae.     

Loss of nervous system complexity – Morphological analyses shed light on the neuronal evolution in Myzostomida (Annelida)

Myzostomida are putatively nested within the errant Annelida and exhibit a unique morphology. The latter fact might possibly be related to their long host-dependent radiation. Hence, an incomplete segmentation, lack of prominent sensory structures in adults and a dorso-ventrally flattened body are just some examples. Although numerous investigations of the nervous system exist for myzostomids, detailed ultrastructural as well as histological examinations of neuronal structures are lacking so far. Therefore, we investigate the nervous system of Myzostoma cirriferum Leuckart, 1836 using a comparative approach including paraffin histology, serial semi-thin sections, immunohistochemistry and ultrastructural investigations. Our analyses reveal a lack of complexity within the anterior-most neuronal condensation (herein called brain) of adult specimens. Hence, prominent tracts or glomeruli are absent, and a glial layer surrounding the brain or radial-like glial cells are lacking. Nevertheless, the ultrastructure of the ventral nervous system is comparable to other Errantia. Therefore, our investigations hint towards a reduction of neuronal complexity in Myzostomida. Additionally, an ontogenetic simplification seems plausible, although further investigations are necessary to verify such a hypothesis. A simplification of neuronal structures due to a drastic change in lifestyle was so far mainly observed for basally branching annelid clades.     

Testing phylogenetic implications of eggshell characters in side-necked turtles (Testudines: Pleurodira)

Amniote egg and eggshell morphology is a rich source of characters to link aspects of reproductive biology with systematics. Extensive work concerning both anatomy and phylogenetic assignability has been done on fossil bird and dinosaur eggs, but little is known for extant sauropsids. The utility of eggshell characters for phylogenetic analyses is tested and discussed for extant side-necked turtles (Pleurodira), and the diversity of egg ultrastructure is examined in several species. Egg gross morphology and eggshell ultrastructure of 12 species of extant side-necked turtles was documented using scanning electron microscopy. Thirteen eggshell characters were scored and mapped on a composite phylogeny and ancestral character states were reconstructed. Many of the characters do not show a phylogenetic signal according to a test comparing the number of steps on the chosen phylogeny with that on randomly generated trees. The presence of conservative, clade-supporting features could be demonstrated, and the following clades are supported by several characters: the Elseya-Emydura entity, short-necked Australasian chelids, is backed by two characters, and two additional characters could potentially support this group. Three characters support the monophyly of South American chelids, whereas two characters argue for the exclusion of Hydromedusa, a long-necked form resembling Australian chelids rather than South American forms, from this clade.     

Anatomy and ultrastructure of the male reproductive system in Pleioplana atomata (Platyhelminthes: Polycladida)

Abstract. The ultrastructure of the male reproductive system in the polyclad flatworm Pleioplana atomata is described. Numerous testes are scattered throughout the entire body but are heavily concentrated on the ventral side. All stages of differentiating sperm cells are present in all testes follicles. Intercellular bridges connect spermatocytes and spermatids derived from a single spermatogonium. In the distal part of spermatids, a zone of differentiation develops with a row of microtubules beneath the plasmalemma. Adjacent to these microtubules, an intercentriolar body is flanked by two basal bodies that give rise to two axonemes (each with a 9+“1” microtubular pattern) that face in opposite directions. The Golgi complex appears in the central portion of the spermatid and produces numerous small and large electron-dense bodies. The small bodies surround the nucleus, whereas the large bodies cluster along with the mitochondria in the central part of the spermatid. Development of the spermatid leads to cell elongation and formation of a filiform, biflagellate mature spermatozoon with cortical microtubules all along the sperm shaft. The male canal system consists of paired vasa deferentia that separately enter a single seminal vesicle. A single prostatic canal connects the seminal vesicle to the prostatic vesicle. Ultrastructurally, the seminal vesicle and prostatic canal are very similar, and along with the prostatic vesicle and stylet pocket, are lined by a ciliated epithelium. The ultrastructure of the prostatic vesicle indicates that it probably produces a large volume of seminal fluid that, along with spermatozoa, is transferred to the mating partner through a stylet. Some of the findings, particularly on sperm ultrastructure, may provide characters useful for phylogenetic analysis.     

角马蜂贮精囊及精子的形态学观察

昆虫贮精囊和精子的形态多样性是重要的分类和系统发育分析特征之一, 然而在马蜂亚科乃至整个胡蜂科中却鲜有涉及。本文首次解剖了角马蜂Polistes chinensis antennalis Pérez的雄性生殖系统, 着重对其贮精囊的超微结构进行描述, 并简要报道了精子的外部形态。角马蜂的贮精囊由输精管亚前端膨大而成, 有一层发达的柱状上皮细胞贴在基底膜内壁： 细胞核位于柱状细胞基部, 上皮细胞端半部线粒体密集, 顶膜特化成微绒毛。角马蜂精子头长21.4 μm, 体长94 μm, 是已报道胡蜂科精子中长度最短、 相对头长最长的种类。研究结果为胡蜂科昆虫系统发育以及繁殖生理提供理论依据。     

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. J. Morphol. 2009. © 2008 Wiley‐Liss, Inc.     

Humidity levels drive reproductive modes and phylogenetic diversity of amphibians in the Brazilian Atlantic Forest

Aim The diversity of reproductive modes among amphibians constitutes a striking example of how differences in the biology of species provide important explanations for species distribution patterns on a broad scale. We hypothesize that sites with a higher humidity level will support more modes of reproduction than drier sites and will consequently exhibit a higher phylogenetic diversity. Furthermore, if there is a gradient in the tolerance of reproductive modes to desiccation, there will be a nested pattern in the composition of reproductive modes among sites. Location Twenty‐seven forest sites in the Brazilian Atlantic Forest. Methods Through a path analysis approach, we evaluated the direct and indirect effects of the humidity level on the number of reproductive modes, as well as the relative importance of both variables on amphibian phylogenetic diversity. A nestedness analysis was used to quantify the extent to which the compositions of both species and reproductive modes in drier sites correspond to subsets of those in sites with higher annual precipitation. Results We found that the reproductive modes present in drier sites are non‐random subsets of those present in sites with higher humidity levels. Because reproductive modes are phylogenetically conserved among amphibians, sites with a greater number of reproductive modes supported greater phylogenetic diversity. Sites with high precipitation throughout the year provided suitable environmental conditions for a larger number of reproductive modes, whereas sites with low precipitation and typical seasonal climates supported only those reproductive modes specialized to resist desiccation. Main conclusions Our results show that humidity‐related variables are key environmental factors related to both the richness of reproductive modes and phylogenetic diversity. Our results support the hypothesis that the higher phylogenetic diversity found in moister sites reflects differences in the tolerance to desiccation among different reproductive modes. Given that reproductive modes are associated with susceptibility to desiccation, their incorporation into explanatory models may trigger a significant advance in the understanding of the mechanisms regulating the species richness and composition of amphibian communities.     

Chloroplast structure of diatoms of different classes

The diversity of chloroplast forms, and their number and cellular location, as well as pyrenoid structure, distinguishes diatoms from other groups of heterokont algae. The fine chloroplast structure is considered to be informative for taxonomic and phylogenetic studies of diatoms. Six species of diatoms belonging to different classes have been examined with transmission electron microscopy. New data on the chloroplast structure have been obtained. Characteristics of the pyrenoid ultrastructure of diatoms belonging to various phylogenetic clades have been defined more precisely. The results specify the ultrastructure of pyrenoids for different phylogenetic clades of diatoms and contribute to the previously obtained data.     

Comparative sperm ultrastructure in Nemertea

Although the monophyly of Nemertea is strongly supported by unique morphological characters and results of molecular phylogenetic studies, their ingroup relationships are largely unresolved. To contribute solving this problem we studied sperm ultrastructure of 12 nemertean species that belong to different subtaxa representing the commonly recognized major monophyletic groups. The study yielded a set of 26 characters with an unexpected variation among species of the same genus (Tubulanus and Procephalothrix species), whereas other species varied in metric values or only one character state (Ramphogordius). In some species, the sperm nucleus has grooves (Zygonemertes virescens, Amphiporus imparispinosus) that may be twisted and give a spiral shape to the sperm head (Paranemertes peregrina, Emplectonema gracile). To make the characters from sperm ultrastructure accessible for further phylogenetic analyses, they were coded in a character matrix. Published data for eight species turned out to be sufficiently detailed to be included. Comparative evaluation of available information on the sperm ultrastructure suggests that subtaxa of Heteronemertea and Hoplonemertea are supported as monophyletic by sperm morphology. However, the data do not provide information on the existing contradictions regarding the internal relationships of “Palaeonemertea.” Nevertheless, our study provides evidence that sperm ultrastructure yields numerous potentially informative characters that will be included in upcoming phylogenetic analyses. J. Morphol. 2010. © 2010 Wiley‐Liss, Inc.     

Morphology and ultrastructure of the testes and spermatozoa of the South African leopard (Panthera pardus)

The survival of South African leopard (Panthera pardus) populations is at risk due to the decrease in natural habitat caused by increasing human populations, development projects and industry. The use of ex situ conservation methods such as assisted reproductive techniques (ARTs) in captive breeding programmes can be used to conserve and protect this species. However, a good understanding of the reproductive system of the species is imperative in the development of successful ARTs. This study serves to provide information on the morphology and ultrastructure of the spermatozoa and testes of P. pardus that can be used to help in the development, testing and optimization of reproductive techniques. The objectives of this study were (i) to assess the morphology and ultrastructure of the spermatozoa by means of fluorescence and electron microscopy respectively, and (ii) to examine the morphology and ultrastructure of the testicular tissue as well as of the stages of spermatogenesis using light and electron microscopy. In this study, it was noted that the morphology and ultrastructure of the spermatozoa are similar to those reported in other feline and mammalian species and that a high amount of morphologically abnormal ejaculated sperm was found to be a common occurrence in all samples obtained.     

Morphology and function of reproductive organs in Neodasys chaetonotoideus (Gastrotricha: Neodasys) with a phylogenetic assessment of the reproductive system in Gastrotricha

The reproductive system of the important basal gastrotrich Neodasys chaetonotoideus is described and reconstructed on the basis of light microscopy, serial ultrathin sections (ultrastructure) and scanning electron microscopy. Starting frontally, the hermaphroditic reproductive system consists of paired and tube shaped lateral testes that do not possess elongated seminal ducts but most likely open directly via paired ventral pores. The unpaired, medio‐dorsal ovary region contains early oogenic stages that mature caudally towards the uterus region, where the most mature egg is positioned laterally to the midgut. The ovary region is not covered with an epithelial lining whereas the uterus region possesses a distinct epithelial wall. Between ovary and uterus region, we have detected a conspicuous section of the female gonad, the vitellogenic oviduct that consists of a thick epithelial wall which forms cellular protuberances into the developing oocytes passing the oviduct. We interpret this as a special, hitherto undescribed mode of vitellogenesis in Gastrotricha. Further caudally, the uterus continues with the fronto‐caudal organ, a complex of two substructures that are apparently homologous to the frontal organ and the caudal organ of many species of the Gastrotricha Macrodasyida. Neodasys chaetonotoideus obviously engages in spermatophore formation and transfer. In this study we develop a morpho‐functional scenario for the gonads and accessory organs in terms of spermatophore production, exchange and oviposition. We compare our newly obtained data with already published results on the reproductive organs of several species of Gastrotricha by means of a species‐character matrix and provide a computer aided evaluation by a parsimonious character optimization. A reconstruction of the reproductive system of the stem species of Gastrotricha on the basis of three recent phylogenetic analyses is presented. These reconstructions give support for a Neodasys‐like reproductive system in the ground pattern of Gastrotricha with slight morphological differences and direct transfer of spermatozoa rather than spermatophore transfer. The evolution of selected characters is traced thus revealing some incidents of convergent evolution as well as the evolutionary replacement of the ancestral frontal organ by the derived frontal sac in at least two separated lineages.