Nemertean Research Articles

Levels and Profile of Tetrodotoxins in Spawning Cephalothrix mokievskii (Palaeonemertea, Nemertea): Assessing the Potential Toxic Pressure on Marine Ecosystems.

The ribbon worms of the closely related species Cephalothrix simula, Cephalothrix cf. simula, and Cephalothrix mokievskii, representing the C. simula species complex, possess high concentrations of tetrodotoxin (TTX) and its analogues in all developmental stages from eggs to adults. It has recently been suggested that the eggs and larvae of these animals can be a source of tetrodotoxins (TTXs) for other aquatic organisms. In the current study, TTXs in mature and post-spawning individuals and in the eggs of C. mokievskii were identified using high-performance liquid chromatography-tandem mass spectrometry. For the first time, the quantity and profile of TTXs that these nemerteans released into the environment during spawning were estimated. We showed that the spawning C. mokievskii females released significant amounts of TTX and 5,6,11-trideoxyTTX with their eggs; these levels were sufficient for the potential toxification of marine bioresources. The issues surrounding the monitoring of TTXs in commercial marine animals, and collecting at the sites of the spawning of nemerteans from the C. simula species complex, are discussed.

Proteotranscriptomic Profiling of the Toxic Mucus of Kulikovia alborostrata (Pilidiophora, Nemertea).

Nemertea is a phylum of bilaterally symmetrical, coelomate, unsegmented worms, also known as ribbon worms. Most species of the phylum Nemertea are marine predators that contain toxins in the single-celled glands of the proboscis and/or integument. Recent transcriptomic studies have shown that nemerteans from all taxonomic groups possess a wide range of putative protein and peptide toxins, while the proteomic data for these animals are highly limited. In this study, proteotranscriptomic analysis was used to investigate the major protein components of the poison of the nemertean Kulikovia alborostrata. We identified 146 transcripts of putative toxins in the transcriptome of K. alborostrata and five putative toxins among the secreted proteins and peptides of the mucus of the animal. The expression levels of cysteine-rich peptides found in the mucus with similarity to known toxins were evaluated in different parts of the body of the worm by quantitative real-time PCR. The high level of expression of investigated peptides in the integument indicate the protective function of these toxins. Overall, this supports the idea that the mucus of nemerteans is a valuable source of peptide and protein toxins.

Identification of Six Novel Proteins Containing a ZP Module from Nemertean Species.

During fertilization, a series of reactions between the eggs and spermatozoa proceed predominantly in a species-specific manner. The molecules mediating these species-specific reactions remain unknown except in a few organisms. In this study, we focused on two species belonging to the phylum Nemertea, Kulikovia alborostrata and K. fulva, and explored molecules involved in species-specific interactions between gametes. Orthologs of molecules known to be involved in species-specific reactions were not expressed in the ovaries of these two species. In contrast, we identified six novel proteins, named NeZPL1-NeZPL6, containing a ZP module. Among these, we found that NeZPL6 is located on the surface of an unfertilized egg and is suggested to be involved in its interaction with spermatozoa. Furthermore, we found an indel of three amino acids in the EGF-like domain of NeZPL6, which possibly confers species specificity to this interaction. Our results suggested the existence of a novel system for species recognition in animal gametes.

Investigating Non-Native Ribbon Worm Cephalothrix simula as a Potential Source of Tetrodotoxin in British Bivalve Shellfish.

Tetrodotoxin (TTX) is a potent marine neurotoxin found in several phylogenetically diverse organisms, some of which are sought as seafood. Since 2015, TTX has been reported in bivalve shellfish from several estuarine locations along the Mediterranean and European Atlantic coasts, posing an emerging food safety concern. Although reports on spatial and temporal distribution have increased in recent years, processes leading to TTX accumulation in European bivalves are yet to be described. Here, we explored the hypothesis that the ribbon worm species Cephalothrix simula, known to contain high levels of TTX, could play a role in the trophic transfer of the toxin into shellfish. During a field study at a single location in southern England, we confirmed C. simula DNA in seawater adjacent to trestle-farmed Pacific oysters Magallana gigas (formerly Crassostrea gigas) with a history of TTX occurrence. C. simula DNA in seawater was significantly higher in June and July during the active phase of toxin accumulation compared to periods of either no or continually decreasing TTX concentrations in M. gigas. In addition, C. simula DNA was detected in oyster digestive glands collected on 15 June 2021, the day with the highest recorded C. simula DNA abundance in seawater. These findings show evidence of a relationship between C. simula and TTX occurrence, providing support for the hypothesis that bivalves may acquire TTX through filter-feeding on microscopic life forms of C. simula present in the water column at particular periods each year. Although further evidence is needed to confirm such feeding activity, this study significantly contributes to discussions about the biological source of TTX in European bivalve shellfish.

Network analyses on photographic surveys reveal that invertebrate predators do not structure epibenthos in the deep (~2000m) rocky Powell Basin, Weddell Sea, Antarctica

Predator-prey interactions in marine ecosystems control population sizes, maintain species richness, and provide intermediate disturbance. Such ecosystem structuring interactions may be rare in Antarctic epibenthic communities, which are unique among marine ecosystems worldwide for their dominance of soft bodied fauna (sponges, soft and hard corals, and echinoderms) and a simultaneous paucity of shell crushing predators (sharks, rays and durophagous decapods). In the shallow benthos, instead of durophagy, important Antarctic predators such as starfish, pycnogonids (sea spiders), nemertean worms, and nudibranchs employ grazing, scavenging, or sucking strategies. Far less is known about deep sea (>1000 m) Antarctic benthic communities due to the challenging nature of polar data collection, so that photographic surveys provide one of the only means of making in situ observations of these deep sea communities. We used seabed photographs of the deep (~2000m) slope of the Powell Basin, northwest Weddell Sea, taken by the Ocean Floor Observation and Bathymetry System on board the RV Polarstern (PS118, April 2019) to investigate the epibenthic community composition, and Bayesian Network Inference (BNI) to determine the ecological network, namely the ecological associations, including potential invertebrate predator-prey relationships between taxa. Photographs show that the rocky substrates of the basin slope support between 10-22 morphotaxa per photo, and highly abundant communities (density between 106 to 553 individuals/m2). BNI results reveal a network of associations between the sessile and mobile suspension and filter feeding organisms and their physical environment. However, associations between invertebrate predators like starfish, and other organisms, were not detected in the network. This lack of inclusion within the network suggests that, despite the presence of these normally important mobile predators, invertebrate predator-prey interactions on the rocky Powell Basin slope do not have the same ecosystem-regulating impact that they do on shallow Antarctic epibenthic communities.

First Chromosome-Level Genome Assembly of a Ribbon Worm from the Hoplonemertea Clade, Emplectonema gracile, and Its Structural Annotation.

Genome-wide information has so far been unavailable for ribbon worms of the clade Hoplonemertea, the most species-rich class within the phylum Nemertea. While species within Pilidiophora, the sister clade of Hoplonemertea, possess a pilidium larval stage and lack stylets on their proboscis, Hoplonemertea species have a planuliform larva and are armed with stylets employed for the injection of toxins into their prey. To further compare these developmental, physiological, and behavioral differences from a genomic perspective, the availability of a reference genome for a Hoplonemertea species is crucial. Such data will be highly useful for future investigations toward a better understanding of molecular ecology, venom evolution, and regeneration not only in Nemertea but also in other marine invertebrate phyla. To this end, we herein present the annotated chromosome-level genome assembly for Emplectonema gracile (Nemertea; Hoplonemertea; Monostilifera; Emplectonematidae), an easily collected nemertean well suited for laboratory experimentation. The genome has an assembly size of 157.9 Mb. Hi-C scaffolding yielded chromosome-level scaffolds, with a scaffold N50 of 10.0 Mb and a score of 95.1% for complete BUSCO genes found as a single copy. Annotation predicted 20,684 protein-coding genes. The high-quality reference genome reaches an Earth BioGenome standard level of 7.C.Q50.

Ribbon worms (phylum Nemertea) from Bodega Bay, California: A largely undescribed diversity.

The diversity of nemerteans along the Pacific coast of the United States is regarded as well characterized, but there remain many cryptic, undescribed, and "orphan" species (those known only in their larval form). Recent sampling of nemerteans in Oregon and Washington has begun to fill in these taxonomic gaps, but nemertean diversity in California has received relatively little attention over the past 60 years. During the summers of 2019 and 2020, nemertean specimens were collected from 20 locations in the Bodega Bay region of northern California, USA, including rocky intertidal shores, sandy beaches, mudflats, and other habitats. Based on morphological assessment and DNA sequence analysis (partial Cytochrome Oxidase I and 16S rRNA genes), our surveys identified 34 nemertean species. Only 13 of these (38%) can be confidently assigned to described species. Another 11 represent species that are new to science, including members of the genera Riserius, Nipponnemertes, Poseidonemertes, Zygonemertes, Nemertellina, Oerstedia, and three species of uncertain affiliation. The remaining ten species include undescribed or cryptic species of uncertain status that have been found previously along the Pacific Coast of North America. Our surveys also document extensions of known geographic ranges for multiple species, including the first records in California of Antarctonemertesphyllospadicola, Cephalothrixhermaphroditica, and Maculauraoregonensis. This is the first report of the genus Nemertellina in the northeast Pacific and Riserius in California. Overall, our findings highlight how much remains to be learned about the diversity and distribution of nemerteans in the northeast Pacific.

Postembryonic development and lifestyle shift in the commensal ribbon worm

BackgroundVarious morphological adaptations are associated with symbiotic relationships between organisms. One such adaptation is seen in the nemertean genus Malacobdella. All species in the genus are commensals of molluscan hosts, attaching to the surface of host mantles with a terminal sucker. Malacobdella possesses several unique characteristics within the order Monostilifera, exhibiting the terminal sucker and the absence of eyes and apical/cerebral organs, which are related to their adaptation to a commensal lifestyle. Nevertheless, the developmental processes that give rise to these morphological characteristics during their transition from free-living larvae to commensal adults remain uncertain.ResultsIn the present study, therefore, we visualized the developmental processes of the internal morphologies during postembryonic larval stages using fluorescent molecular markers. We demonstrated the developmental processes, including the formation of the sucker primordium and the functional sucker. Furthermore, our data revealed that sensory organs, including apical/cerebral organs, formed in embryonic and early postembryonic stages but degenerated in the late postembryonic stage prior to settlement within their host using a terminal sucker.ConclusionsThis study reveals the formation of the terminal sucker through tissue invagination, shedding light on its adhesion mechanism. Sucker muscle development likely originates from body wall muscles. Notably, M. japonica exhibits negative phototaxis despite lacking larval ocelli. This observation suggests a potential role for other sensory mechanisms, such as the apical and cerebral organs identified in the larvae, in facilitating settlement and adhesive behaviors. The loss of sensory organs during larval development might reflect a transition from planktonic feeding to a stable, host-associated lifestyle. This study also emphasizes the need for further studies to explore the phylogenetic relationships within the infraorder Amphiporiina and investigate the postembryonic development of neuromuscular systems in closely related taxa to gain a more comprehensive understanding of ecological adaptations in Nemertea.

Peptide Toxins from Antarctica: The Nemertean Predator and Scavenger Parborlasia corrugatus (McIntosh, 1876).

Peptide toxins from marine invertebrates have found use as drugs and in biotechnological applications. Many marine habitats, however, remain underexplored for natural products, and the Southern Ocean is among them. Here, we report toxins from one of the top predators in Antarctic waters: the nemertean worm Parborlasia corrugatus (McIntosh, 1876). Transcriptome mining revealed a total of ten putative toxins with a cysteine pattern similar to that of alpha nemertides, four nemertide-beta-type sequences, and two novel full-length parborlysins. Nemertean worms express toxins in the epidermal mucus. Here, the expression was determined by liquid chromatography combined with mass spectrometry. The findings include a new type of nemertide, 8750 Da, containing eight cysteines. In addition, we report the presence of six cysteine-containing peptides. The toxicity of tissue extracts and mucus fractions was tested in an Artemia assay. Notably, significant activity was observed both in tissue and the high-molecular-weight mucus fraction, as well as in a parborlysin fraction. Membrane permeabilization experiments display the membranolytic activity of some peptides, most prominently the parborlysin fraction, with an estimated EC50 of 70 nM.

Tetrodotoxins in Tissues and Cells of Different Body Regions of Ribbon Worms Kulikovia alborostrata and K. manchenkoi from Spokoynaya Bay, Sea of Japan.

Nemerteans, or ribbon worms, possess tetrodotoxin and its analogues (TTXs), neurotoxins of bacterial origin, which they presumably use for capturing prey and self-defense. Most TTXs-containing nemertean species have low levels of these toxins and, therefore, have usually been neglected in studies of TTXs functions and accumulation. In the present study, Kulikovia alborostrata and K. manchenkoi, two closely related species, were analyzed for TTXs distribution in the body using the HPLC-MS/MS and fluorescence microscopy methods. The abundance of TTXs-positive cells was determined in the proboscis, integument, and digestive system epithelium. As a result, six TTXs-positive cell types were identified in each species; however, only four were common. Moreover, the proportions of the toxins in different body parts were estimated. According to the HPLC-MS/MS analysis, the TTXs concentrations in K. alborostrata varied from 0.91 ng/g in the proboscis to 5.52 ng/g in the precerebral region; in K. manchenkoi, the concentrations ranged from 7.47 ng/g in the proboscis to 72.32 ng/g in the posterior body region. The differences observed between the two nemerteans in the distribution of the TTXs were consistent with the differences in the localization of TTXs-positive cells. In addition, TTXs-positive glandular cell types were found in the intestine and characterized for the first time. TTXs in the new cell types were assumed to play a unique physiological role for nemerteans.

New Pyridyl and Dihydroisoquinoline Alkaloids Isolated from the Chevron Nemertean Amphiporus angulatus.

Nemertean worms contain toxins that are used to paralyze their prey and to deter potential predators. Hoplonemerteans often contain pyridyl alkaloids like anabaseine that act through nicotinic acetylcholine receptors and crustacean chemoreceptors. The chemical reactivity of anabaseine, the first nemertean alkaloid to be identified, has been exploited to make drug candidates selective for alpha7 subtype nAChRs. GTS-21, a drug candidate based on the anabaseine scaffold, has pro-cognitive and anti-inflammatory actions in animal models. The circumpolar chevron hoplonemertean Amphiporus angulatus contains a multitude of pyridyl compounds with neurotoxic, anti-feeding, and anti-fouling activities. Here, we report the isolation and structural identification of five new compounds, doubling the number of pyridyl alkaloids known to occur in this species. One compound is an isomer of the tobacco alkaloid anatabine, another is a unique dihydroisoquinoline, and three are analogs of the tetrapyridyl nemertelline. The structural characteristics of these ten compounds suggest several possible pathways for their biosynthesis.

Comparative development of the serotonin- and FMRFamide-immunoreactive components of the nervous system in two distantly related ribbon worm species (Nemertea, Spiralia).

Neurodevelopment in larval stages of non-model organisms, with a focus on the serotonin- and FMRFamide-immunoreactive components, has been in the focus of research in the recent past. However, some taxonomic groups remain understudied. Nemertea (ribbon worms) represent such an understudied clade with only few reports on nervous system development mostly from phylogenetically or developmentally derived species. It would be insightful to explore neurodevelopment in additional species to be able to document the diversity and deduce common patterns to trace the evolution of nervous system development. Fluorescent immunohistochemical labeling with polyclonal primary antibodies against serotonin and FMRF-amide and a monoclonal antibody against synapsin performed on series of fixed larval stages of two nemertean species Cephalothrix rufifrons (Archinemertea, Palaeonemertea) and Emplectonema gracile (Monostilifera, Hoplonemertea) were analyzed with confocal laser scanning microscopy. This contribution gives detailed accounts on the development of the serotonin- and FMRFamide-immunoreactive subsets of the nervous system in two nemertean species from the first appearance of the respective signals. Additionally, data on synapsin-like immunoreactivity illustrates the general structure of neuropil components. Events common to both investigated species are the appearance of serotonin-like immunoreactive signals before the appearance of FMRF-like immunoreactive signals and the strict progression of the development of the lateral nerve cords from the anteriorly located, ring-shaped brain toward the posterior pole of the larva. Notable differences are (1) the proboscis nervous system that is developing much earlier in investigated larval stages of E. gracile and (2) distinct early, but apparently transient, serotonergic neurons on the frontal and caudal pole of the larva in E. gracile that seem to be absent in C. rufifrons. According to the results from this investigation and in line with previously published accounts on nervous system development, the hypothetical last common ancestor of Nemertea had a ring-shaped brain arranged around the proboscis opening, from which a pair of ventro-lateral nerve cords develops in anterior to posterior progression. Early frontal and caudal serotonergic neurons that later degenerate or cease to express serotonin are an ancestral character of Nemertea that they share with several other spiralian clades.

Tetrodotoxin and Its Analogues (TTXs) in the Food-Capture and Defense Organs of the Palaeonemertean Cephalothrix cf. simula.

Tetrodotoxin (TTX), an extremely potent low-molecular-weight neurotoxin, is widespread among marine animals including ribbon worms (Nemertea). Previously, studies on the highly toxic palaeonemertean Cephalothrix cf. simula showed that toxin-positive structures are present all over its body and are mainly associated with glandular cells and epithelial tissues. The highest TTXs concentrations were detected in a total extract from the intestine of the anterior part of the body and also in a total extract from the proboscis. However, many questions as to the TTXs distribution in the organs of the anterior part of the worm's body and the functions of the toxins in these organs are still unanswered. In the present report, we provide additional results of a detailed and comprehensive analysis of TTXs distribution in the nemertean's proboscis, buccal cavity, and cephalic gland using an integrated approach including high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), confocal laser scanning microscopy with anti-TTX antibodies, light and electron microscopies, and observations of feeding behavior. For the proboscis, we have found a TTXs profile different from that characteristic of other organs and tissues. We have also shown for the first time that the major amount of TTXs is localized in the anterior part of the proboscis that is mainly involved in hunting. TTX-containing glandular cells, which can be involved in the prey immobilization, have been found in the buccal cavities of the nemerteans. A significant contribution of the cephalic gland to the toxicity of this animal has been shown for the first time, and the role of the gland is hypothesized to be involved not only in protection against potential enemies but also in immobilizing prey. The data obtained have made it possible to extend the understanding of the role and features of the use of TTXs in the organs of the anterior part of nemertean's body.

Upside-down behaviour of certain ribbon worms (phylum Nemertea)

Abstract Ribbon worms in the genus Balionemertes from Vietnam, the Philippines, Australia, and Guam—as well as Cephalothrix suni from Vietnam—were examined. Our observations indicate that the worms crawl mostly with their ventral surface upwards (the ventral surface being where the mouth opens), a behaviour that has not been documented in previous literature. Like many other worm species with colour patterns, they have a darker-coloured and/or more intensely patterned behavioural dorsal surface (= anatomical ventral surface in Balionemertes and C. suni) than the other side. This type of behavioural dorsoventral body-axis inversion among vermiform benthos seems to be rare—not having hitherto been known at least in the phylum Nemertea—and may be related to their feeding strategy, which should be observed in future studies.

Prevalence, intensity of infestation, and biomarker potential of the nemertean worm, Carcinonemertes carcinophila (Kölliker, 1845), on ovigerous blue crabs, Callinectes sapidus Rathbun, 1896 (Decapoda: Brachyura: Portunidae), in Chesapeake Bay, USA

Abstract The effect of individual and population-level egg mortality is important to quantify to maintain sustainable crustacean fisheries. The nemertean worm Carcinonemertes carcinophila (Kölliker, 1845) is an egg predator of the Atlantic blue crab, Callinectes sapidusRathbun, 1896; however, little is known about the impact this nemertean has on the reproduction of the blue crab. We assessed the prevalence and intensity of the infestation of nemerteans in ovigerous blue crabs using a fishery-independent trawl survey. During the primary spawning period of the crab, May–September 2022, 126 ovigerous females were collected and analyzed for worms. Prevalence over this time was 66.6% and mean brood infestation was 53.9 worms per infested crab host. Nemertean egg consumption was quantified with a six-day microcosm experiment. Of the 48 worms in the experiment, 71% actively fed on crab eggs and their consumption ranged 0.16–4.5 eggs day–1. Consumption rates were used to estimate population-level impact of nemertean feeding on crab brood mortality. Modeled proportions of brood loss per crab ranged 0–0.0044%. At the current prevalence and intensity of infestation, egg consumption by nemerteans has a negligible effect on blue crab reproductive output and batch fecundity in Chesapeake Bay. We also investigated the use of mature nemertean worms as a biomarker for establishing the spawning history of ovigerous female blue crabs and determined that the presence of worms in the clutch and in the gills can be used to indicate parity in ovigerous female crabs.

Venomous Noodles: The Evolution of Toxins in Nemertea through Positive Selection and Gene Duplication.

Some, probably most and perhaps all, members of the phylum Nemertea are poisonous, documented so far from marine and benthic specimens. Although the toxicity of these animals has been long known, systematic studies on the characterization of toxins, mechanisms of toxicity, and toxin evolution for this group are scarce. Here, we present the first investigation of the molecular evolution of toxins in Nemertea. Using a proteo-transcriptomic approach, we described toxins in the body and poisonous mucus of the pilidiophoran Lineus sanguineus and the hoplonemertean Nemertopsis pamelaroeae. Using these new and publicly available transcriptomes, we investigated the molecular evolution of six selected toxin gene families. In addition, we also characterized in silico the toxin genes found in the interstitial hoplonemertean, Ototyphlonemertes erneba, a meiofaunal taxa. We successfully identified over 200 toxin transcripts in each of these species. Evidence of positive selection and gene duplication was observed in all investigated toxin genes. We hypothesized that the increased rates of gene duplications observed for Pilidiophora could be involved with the expansion of toxin genes. Studies concerning the natural history of Nemertea are still needed to understand the evolution of their toxins. Nevertheless, our results show evolutionary mechanisms similar to other venomous groups.

Seven new species of Tetranemertes Chernyshev, 1992 (Monostilifera, Hoplonemertea, Nemertea) from the Caribbean Sea, western Pacific, and Arabian Sea, and revision of the genus.

The marine ribbon worm genus Tetranemertes Chernyshev, 1992 currently includes three species: the type species T.antonina (Quatrefages, 1846) from the Mediterranean Sea, T.rubrolineata (Kirsteuer, 1965) from Madagascar, and T.hermaphroditica (Gibson, 1982) from Australia. Seven new species are described: T.bifrostsp. nov., T.ocelatasp. nov., T.majinbuuisp. nov., and T.pastafariensissp. nov. from the Caribbean Sea (Panamá), and three species, T.unistriatasp. nov., T.paulayisp. nov., and T.arabicasp. nov., from the Indo-West Pacific (Japan and Oman). As a result, an amended morphological diagnosis of the genus is offered. To improve nomenclatural stability, a neotype of Tetranemertesantonina is designated from the Mediterranean. The newly described species, each characterized by features of external appearance and stylet apparatus, as well as by DNA-barcodes, form a well-supported clade with T.antonina on a molecular phylogeny of monostiliferan hoplonemerteans based on partial sequences of COI, 16S rRNA, 18S rRNA, and 28S rRNA. Six of the seven newly described species, as well as T.rubrolineata, possess the unusual character of having a central stylet basis slightly bilobed to deeply forked posteriorly in fully grown individuals, a possible morphological synapomorphy of the genus. In addition, an undescribed species of Tetranemertes is reported from the Eastern Tropical Pacific (Panamá), increasing the total number of known species in the genus to eleven.

Tetrodotoxins in Larval Development of Ribbon Worm Cephalothrix cf. simula (Palaeonemertea, Nemertea).

The toxic ribbon worm, Cephalothrix cf. simula (Palaeonemertea, Nemertea), possesses extremely high concentrations of tetrodotoxin (TTX). Although TTX has been found in the eggs of this species, the fate of the toxin in the ontogeny of the animal has not been explored. Here, using high performance liquid chromatography with tandem mass spectrometry and immunohistochemistry with anti-TTX antibodies, we examined levels, profile, and localization of TTX and its analogues (TTXs) in larvae of C. cf. simula throughout 41days post-fertilization. A detailed investigation of cells in sites of TTX-accumulation was performed with light and electron microscopy. Newly hatched larvae possessed weak TTX-like immunoreactivity in all cells. With subsequent development, intensity of TTX-labeling in the ectodermal structures, mesodermal cells and apical cylinder of the apical gland increased. In the ectodermal structures, an intense TTX-labeling was observed in the multiciliated, type II granular, type I mucoid, and basal cells of the epidermis, and in the type III granular cells of the mouth gland. In the mesoderm, TTX was localized in the muscle and unigranular parenchyma-like cells. Eggs and larvae of C. cf. simula contained five TTXs, with two major toxins - TTX and 5,6,11-trideoxyTTX. Level and relative proportion of TTXs did not differ significantly among developmental stages, confirming that larvae obtained toxins from maternal eggs and were able to retain it. The results of this study provide insights into the formation of TTX-bearing apparatus of C. cf. simula through the larval development.

The effect of the egg-predator Carcinonemertes conanobrieni on the reproductive performance of the Caribbean spiny lobster Panulirus argus

BackgroundThe Caribbean spiny lobster Panulirus argus is heavily fished throughout its Greater Caribbean and Gulf of Mexico distribution, suggesting a heightened susceptibility to a fisheries collapse. In 2017, a nemertean worm, Carcinonemertes conanobrieni was described from ovigerous females of P. argus in Florida, USA. A year later, the presence of the same egg predator was recorded along the southern Caribbean coast (Colombia). The effect of this egg predator on the reproductive performance, including fecundity, embryo mortality, and reproductive output, of its host is unknown. This study tested whether C. conanobrieni affects embryo mortality, fecundity, and reproductive output in brooding females of P. argus.ResultsArtisan fishers caught 90 ovigerous lobsters near Pueblo Viejo, Magdalena, Colombia. Each ovigerous female was examined for the presence/absence of the egg predator. Lobster egg mortality (%), fecundity (nº eggs female−1), and reproductive output (%) were estimated. Prevalence of C. conanobrieni in the studied population was 87.78%. The mean intensity of C. conanobrieni (all life stages) in the population was 11.68 (± 1.98) egg predators per brood mass sample. Infected females brooding late-stage embryos exhibited lower fecundity, lower reproductive performance values, and higher embryo mortality compared to infected females brooding early-stage embryos. Embryo stage and worm infection level negatively impacted fecundity and reproductive output. Worm infection level and the number of adult nemertean worms also negatively affected embryo mortality.ConclusionsThese results demonstrate an adverse effect of C. conanobrieni on the reproductive performance of P. argus. The interactive impact of this egg predator, natural stressors, and anthropogenic stressors on individual P. argus reproductive performance could facilitate losses at large-scale fisheries levels.

At the edge of the sea: the supralittoral nemertean, Acteonemertes orientalis sp. nov. (Nemertea: Eumonostilifera: Plectonemertidae) from Japan

Ribbon worms (phylum Nemertea) are found in oceans worldwide; however, only a few inhabit terrestrial, semiterrestrial and freshwater environments. In our study, we describe Acteonemertes orientalis sp. nov., a new species of Plectonemertidae Gibson, 1990 and the first plectonemertid discovered in Japan. The species was found in the supralittoral zone, from which nemerteans have rarely been reported and inhabits areas under rocks, wave-dissipating concrete blocks, and fallen leaves and stocks, along the coast of the Sea of Japan. Multi-locus phylogenetic analyses based on two mitochondrial (16S rRNA and cytochrome c oxidase subunit I) and two nuclear gene markers (18S rRNA and 28S rRNA) revealed that A. orientalis sp. nov. formed a clade with Leptonemertes cf. chalicophora collected from Florida. Although the external and internal morphology of A. orientalis sp. nov. is largely consistent with the generic diagnosis, this species differs in having a large proboscis. Based on the molecular and morphological data, we propose to amend the diagnosis of Acteonemertes by excluding proboscis size. ZooBank: urn:lsid:zoobank.org:pub:BBBE9549-5DCE-424F-9A39-930F8243C28B