Discovery and characterization of a transient chaetal gland during the development of Capitella teleta (Sedentaria: Annelida).

Chaetae are among the most extensively studied structures in polychaetes, serving as a defining morphological trait for annelids. Capitella teleta stands out as one of the few established annelid models for developmental and morphological studies, thus receiving significant scholarly attention. In this study, we unveil a previously unnoticed glandular structure associated with chaetae within the larvae of C. teleta. Our investigations demonstrate the absence of comparable structures in the chaetal follicles of adults and juveniles (older than 1 week), as well as during active chaetogenesis, underscoring the transient nature of these glands. This indicates that larval chaetal follicles transform into a gland that later disappears. Utilizing histology and transmission electron microscopy, we characterized these glands. Our findings underscore the diversity of chaetal ultrastructure in annelids and show that, even in well-studied species, novel morphological details can be found. We emphasize the importance of examining various life-history stages to capture such transient morphological features. This work lays a crucial morphological foundation and deepens our understanding of chaetae and chaetogenesis in C. teleta, paving the way for more accurate interpretations of future experimental studies on chaetogenesis in this species.

Morphometric and taxonomic approach to describe Heterospio variabilis (Annelida, Longosomatidae), a new species with three size-dependent morphotypes, from the Gulf of California, Eastern Pacific.

The Longosomatidae, a poorly known polychaete family, includes only 23 recognized species; in this study, based on morphometric and taxonomic analyses, we describe a new species with three morphotypes: Heterospio variabilis from the Gulf of California, Mexico. The specimens examined exhibit large morphological variations but were clearly separated from close species due to a unique combination of morphological characters: chaetiger 9 as the first elongated chaetiger, four to eight branchial pairs; chaetae from chaetiger 10 forming rings in two rows, posterior row with thin and robust capillaries, anterior row with subuluncini, aristate spines, acicular spines and thick acicular spines. With the discriminant analysis, carried out on 11 morphometric characters, the presence of three morphological groups were recognized (Wilks' lambda= 0.093, p = 0.0001). However, the variables selected to discriminate the specimens (partial Wilks' lambda > 0.57) were correlated to their size: number of branchiae, body width, prostomium width, rate length CH9/CH1-CH8, length CH1-CH8 and length CH9 (r > 0.5). So, we concluded that they belong to a single species with three morphotypes: morpho A with eight branchial pairs, morpho B with 5-6-7 pairs and morpho C with 4 pairs. No correlations between the distribution of the distinct morphotypes along the eastern gulf shelf and the environmental conditions where they settle were detected.

Is there potential for estradiol receptor signaling in lophotrochozoans?

Estrogen receptors (ERs) are thought to be the ancestor of all steroid receptors and are present in most lophotrochozoans studied to date, including molluscs, annelids, and rotifers. A number of studies have investigated the functional role of estrogen receptors in invertebrate species, although most are in molluscs, where the receptor is constitutively active. In vitro experiments provided evidence for ligand-activated estrogen receptors in annelids, raising important questions about the role of estrogen signalling in lophotrochozoan lineages. Here, we review the concordant and discordant evidence of estradiol receptor signalling in lophotrochozoans, with a focus on annelids and rotifers. We explore the de novo synthesis of estrogens, the evolution and expression of estrogen receptors, and physiological responses to activation of estrogen receptors in the lophotrochozoan phyla Annelida and Rotifera. Key data are missing to determine if de novo biosynthesis of estradiol in non-molluscan lophotrochozoans is likely. For example, an ortholog for the CYP11 gene is present, but confirmation of substrate conversion and measured tissue products is lacking. Orthologs CYP17 and CYP19 are lacking, yet intermediates or products (e.g. estradiol) in tissues have been measured. Estrogen receptors are present in multiple species, and for a limited number, in vitro data show agonist binding of estradiol and/or transcriptional activation. The expression patterns of the lophotrochozoan ERs suggest developmental, reproductive, and digestive roles but are highly species dependent. E2 exposures suggest that lophotrochozoan ERs may play a role in reproduction, but no strong dose-response relationship has been established. Therefore, we expect most lophotrochozoan species, outside of perhaps platyhelminths, to have an ER but their physiological role remains elusive. Mining genomes for orthologs gene families responsible for steroidogenesis, coupled with in vitro and in vivo studies of the steroid pathway are needed to better assess whether lophotrochozoans are capable of estradiol biosynthesis. One major challenge is that much of the data are divided across a diversity of species. We propose that the polychaetes Capitella teleta or Platyneris dumerilii, and rotifer Brachionus manjavacas may be strong species choices for studies of estrogen receptor signalling, because of available genomic data, established laboratory culture techniques, and gene knockout potential.

Highly Concentrated Linear Guanidine Amides from the Marine Sipunculid Phascolosoma granulatum.

The chemical diversity of annelids, particularly those belonging to the class Sipuncula, remains largely unexplored. However, as part of a Marine Biodiscovery program in Ireland, the peanut worm Phascolosoma granulatum emerged as a promising source of unique metabolites. The purification of the MeOH/CH2Cl2 extract of this species led to the isolation of six new linear guanidine amides, named phascolosomines A-F (1-6). NMR analysis allowed for the elucidation of their structures, all of which feature a terminal guanidine, central amide linkage, and a terminal isobutyl group. Notably, these guanidine amides were present in unusually high concentrations, comprising ∼3% of the dry mass of the organism. The primary concentration of the phascolosomines in the viscera is similar to that previously identified in linear amides from sipunculid worms and marine fireworms. The compounds from sipunculid worms have been hypothesized to be toxins, while those from fireworms are reported to be defensive irritants. However, screening of the newly isolated compounds for inhibitory bioactivity showed no significant inhibition in any of the assays conducted.

Coinfection of slime feather duster worms (Annelida, Myxicola) by different gregarine apicomplexans (Selenidium) and astome ciliates reflects spatial niche partitioning and host specificity.

Individual organisms can host multiple species of parasites (or symbionts), and one species of parasite can infect different host species, creating complex interactions among multiple hosts and parasites. When multiple parasite species coexist in a host, they may compete or use strategies, such as spatial niche partitioning, to reduce competition. Here, we present a host­symbiont system with two species of Selenidium (Apicomplexa, Gregarinida) and one species of astome ciliate co-infecting two different species of slime feather duster worms (Annelida, Sabellidae, Myxicola) living in neighbouring habitats. We examined the morphology of the endosymbionts with light and scanning electron microscopy (SEM) and inferred their phylogenetic interrelationships using small subunit (SSU) rDNA sequences. In the host 'Myxicola sp. Quadra', we found two distinct species of Selenidium; S. cf. mesnili exclusively inhabited the foregut, and S. elongatum n. sp. inhabited the mid to hindgut, reflecting spatial niche partitioning. Selenidium elongatum n. sp. was also present in the host M. aesthetica, which harboured the astome ciliate Pennarella elegantia n. gen. et sp. Selenidium cf. mesnili and P. elegantia n. gen. et sp. were absent in the other host species, indicating host specificity. This system offers an intriguing opportunity to explore diverse aspects of host­endosymbiont interactions and competition among endosymbionts.

Four new records of Sipuncula (Annelida) from Northern Brazil.

There are very few taxonomic studies of Brazilian sipunculan worms, and the species occurring along the northern coasts are largely understudied. We report herein the occurrence of four shallow water species along the intertidal zones of Bahia and Pernambuco states. Antillesoma antillarum (Grbe & Oersted, 1858) occurs from underneath beach rocks or within sandstone reefs and may reach high densities. The large species Sipunculus (Sipunculus) nudus Linnaeus, 1766, Sipunculus polymyotus Fisher, 1947, and Xenosiphon branchiatus (Fischer, 1895) were collected in sandy-mud, low-energy intertidal environments. For the first time, we report that sipunculans are collected and used as fishing bait by local fishermen in the Western Atlantic. All species are fully described, including notes on their external and internal anatomy.

First record of Oligobregma Kudenov & Blake, 1975 (Annelida: Polychaeta: Scalibregmatidae Malmgren, 1867) from Brazil with the description of three new species.

Oligobregma represents the most speciose genus of Scalibregmatidae with 17 valid species. Most of them occur at great depths and are found living on soft bottoms. Here, we present the descriptions of O. nonatoi sp. nov., O. cruzae sp. nov., and O. bakkeni sp. nov., sampled from the Brazilian continental shelf and slope, and a key to all species of Oligobregma. O. nonatoi sp. nov. differs from its congeners by the presence of a triangular prostomium with short rounded horns, acicular spines on notopodia of chaetigers 14 and on neuropodia of chaetigers 13. Oligobregma cruzae sp. nov. can be distinguished by the presence of a quadrangular prostomium with two short, rounded horns, acicular spines on chaetigers 14, and lyrate chaetae from chaetiger 5. Finally, O. bakkeni sp. nov. is unique among its congeners by the presence of acicular spines with rounded tips. This is the first record of this genus along the Brazilian coast and the specimens were collected on Esprito Santo, Campos and Santos sedimentary basins, both located in Brazilian southeast region. The sampling campaigns were carried out in scientific expeditions related to two research projects: Santos Project Santos Basin Environmental Characterization and Ambes Project Esprito Santo Basin Assessment Project, both coordinated by CENPES/PETROBRAS.

A reevaluation and redescription of the branchiobdellidan Sathodrilus tetrodonta (Pierantoni, 1906) new comb. (Annelida: Clitellata).

The questionable status of Branchiobdella tetrodonta Pierantoni, 1906 is resolved and the species is transferred to the correct genus. The original description was made from specimens removed from signal crayfish collected in California, USA, unfortunately, Pierantoni (1906c) did not designate any type specimens nor where the preparations were deposited; they are now presumed lost. Holt (1967) believed B. tetrodonta possessed unique penial hooks and a chitinous sheath which was due to his mistranslation of the original Italian description. As a result, Sathodrilus attenuatus Holt, 1981, was described even though specimens came from the same area and host and possessed very similar jaw characteristics to B. tetrodonta. In addition, the jaw characteristics of both endemic species are unique in the Pacific Ocean drainage of the USA. A reassessment of the literature and re-examination of Holts type specimens has resulted in the species name becoming a new combination, Sathodrilus tetrodonta (Pierantoni, 1906), with Sathodrilus attenuatus Holt, 1981, as its junior synonym.

A remarkable new deep-sea nereidid (Annelida: Nereididae) with gills.

Nereidid polychaetes are well known from shallow marine habitats, but their diversity in the deep sea is poorly known. Here we describe an unusual new nereidid species found at methane seeps off the Pacific coast of Costa Rica. Specimens of Pectinereis strickrotti gen. nov., sp. nov. had been observed dating back to 2009 swimming just above the seafloor at ~1,000 m depth but were not successfully captured until 2018. Male epitokes were collected as well as a fragment of an infaunal female found in a pushcore sample. The specimens were all confirmed as the same species based on mitochondrial COI. Phylogenetic analyses, including one based on available whole mitochondrial genomes for nereidids, revealed no close relative, allowing for the placement of the new species in its own genus within the subfamily Nereidinae. This was supported by the unusual non-reproductive and epitokous morphology, including parapodial cirrostyles as pectinate gills, hooked aciculae, elfin-shoe-shaped ventral cirrophores, and elongate, fusiform dorsal ligules emerging sub-medially to enlarged cirrophores. Additionally, the gill-bearing subfamily Dendronereidinae, generally regarded as a junior synonym of Gymnonereidinae, is reviewed and it is here reinstated and as a monogeneric taxon.

The development of the adult nervous system in the annelid Owenia fusiformis.

BACKGROUND: The evolutionary origins of animal nervous systems remain contentious because we still have a limited understanding of neural development in most major animal clades. Annelids - a species-rich group with centralised nervous systems - have played central roles in hypotheses about the origins of animal nervous systems. However, most studies have focused on adults of deeply nested species in the annelid tree. Recently, Owenia fusiformis has emerged as an informative species to reconstruct ancestral traits in Annelida, given its phylogenetic position within the sister clade to all remaining annelids. METHODS: Combining immunohistochemistry of the conserved neuropeptides FVamide-lir, RYamide-lir, RGWamide-lir and MIP-lir with gene expression, we comprehensively characterise neural development from larva to adulthood in Owenia fusiformis. RESULTS: The early larval nervous system comprises a neuropeptide-rich apical organ connected through peripheral nerves to a prototroch ring and the chaetal sac. There are seven sensory neurons in the prototroch. A bilobed brain forms below the apical organ and connects to the ventral nerve cord of the developing juvenile. During metamorphosis, the brain compresses, becoming ring-shaped, and the trunk nervous system develops several longitudinal cords and segmented lateral nerves. CONCLUSIONS: Our findings reveal the formation and reorganisation of the nervous system during the life cycle of O. fusiformis, an early-branching annelid. Despite its apparent neuroanatomical simplicity, this species has a diverse peptidergic nervous system, exhibiting morphological similarities with other annelids, particularly at the larval stages. Our work supports the importance of neuropeptides in animal nervous systems and highlights how neuropeptides are differentially used throughout development.

The Mitochondrial Genomes of Siboglinum plumosum and Oligobrachia dogieli (Annelida: Siboglinidae) and Their Phylogenetic Analysis.

Frenulates are a group of sedentary Annelida within the family Siboglinidae that inhabit the ocean floor and present a unique challenge for comprehensive molecular and phylogenetic investigations. In this study, we focused on the frenulates, specifically assembling the mitochondrial genomes of Siboglinum plumosum and Oligobrachia dogieli. The phylogenetic reconstruction placed S. plumosum as a sister taxon to S. ekmani, and O. dogieli as a sister taxon to S. fiordicum, supporting the non-monophyletic nature of the genus Siboglinum. Overall, this study supports the phylogeny of the family Siboglinidae while highlighting the need for additional molecular data within frenulates.

Discovery and characterization of the α-amylases cDNAs from Enchytraeus albidus shed light on the evolution of "Enchytraeus-Eisenia type" Amy homologs in Annelida.

Although enchytraeids have gained popularity in scientific research, fundamental questions regarding their feeding ecology and biology remain largely unexplored. This study investigates α-amylases, major digestive enzymes responsible for hydrolyzing starch and similar polysaccharides into sugars, in Enchytraeus albidus. Genetic data related to α-amylases is currently lacking for the family Enchytraeidae but also for the entire Annelida. To detect and identify coding sequences of the expressed α-amylase genes in COI-monohaplotype culture (PL-A strain) of E. albidus, we used classical "gene fishing" and transcriptomic approaches. We also compared coding sequence variants of α-amylase retrieved from transcriptomic data related to freeze-tolerant strains. Our results reveal that E. albidus possesses two distinct α-amylase genes (Amy I and Amy II) that are homologs to earthworm Eisenia fetida Ef-Amy genes. Different strains of E. albidus possess distinctive alleles of α-amylases with unique SNP patterns specific to a particular strain. Unlike Amy II, Amy I seems to be a highly polymorphic and multicopy gene. The domain architecture of the putative Amy proteins was found the same as for classical animal α-amylases with ABC-domains. A characteristic feature of Amy II is the lack of GHGA motif in the flexible loop region, similarly to many insect amylases. We identified "Enchytraeus-Eisenia type" α-amylase homologs in other clitellates and polychaetes, indicating the ancestral origin of Amy I/II proteins in Annelida. This study provides the first insight into the endogenous non-proteolytic digestive enzyme genes in potworms, discusses the evolution of Amy α-amylases in Annelida, and explores phylogenetic implications.

New species of Pseudoscalibregma Ashworth, 1901 (Annelida: Scalibregmatidae Malmgren, 1867) from Brazil.

Pseudoscalibregma Ashworth, 1901 is a small genus of the family Scalibregmatidae Malmgren, 1867. Currently, a total of eight valid species are accepted. Of these, the majority are found at great depths. This work contains the first report of Pseudoscalibregma to the Brazilian coast, with the description of three new species: Pseuscalibregma ermindae sp. nov., P. magalhaesi sp. nov. and P. parapari sp. nov. The material was sampled from the Brazilian southeast region, on Campos and Santos sedimentary basins, during scientific expeditions called Santos ProjectSantos Basin Environmental Characterization and Ambes ProjectEsprito Santo Basin Assessment Project, both coordinated by PETROBRAS/CENPES. We also include an identification key to Pseudoscalibregma species.

A single coelomic cell type is involved in both immune and respiratory functions of the coastal bioindicator annelid: Capitella C-Channel1 from the English Channel.

The polychaete Capitella is a typical member of the 'thiobiome', and is commonly used as an eutrophication indicator species in environmental assessment studies. To deal with a sulfide-rich and poisonous surrounding, cells in close contact with the environment, and thus able to play a major role in detoxication and survival, are circulating cells. This work aimed to morpho-functionally describe the circulating coelomic cells of Capitella from the English Channel inhabiting the sulfide-rich mud in Roscoff Harbor. In general, worms have three types of circulating cells, granulocytes involved in bacterial clearance and defense against microorganisms, eleocytes with an essentially trophic role and elimination of cellular waste, and erythrocytes which play a role in detoxification and respiration via their intracellular hemoglobin. By combining diverse microscopic and cellular approaches, we provide evidence that Capitella does not possess granulocytes and eleocytes, but rather a single abundant rounded cell type with the morphological characteristics of erythrocytes i.e. small size and production of intracellular hemoglobin. Surprisingly, our data show that in addition to their respiratory function, these red cells could exert phagocytic activities, and produce an antimicrobial peptide. This latter immune role is usually supported by granulocytes. Our data highlight that the erythrocytes of Capitella from the English Channel differ in morphology and bear more functions than the erythrocytes of other annelids. The simplicity of this multi-task (or polyvalent) single-cell type makes Capitella an interesting model for studies of the impact of the environment on the immunity of this bioindicator species.

Identification and expression analysis of Oxfibrillin gene involved in the regeneration process of Ophryotrocha xiamen (Annelida, Dorcilleidae).

Regeneration of lost body parts is a widespread phenomenon across annelids. However, the molecular inducers of the cell sources for this reparative morphogenesis have not been identified. We have identified a regeneration-related gene Oxfibrillin from the transcriptome analysis of a polychaeta, Ophryotrocha xiamen, which is found to be a well-suited model to study the mechanisms of regeneration. Fibrillins are large glycoproteins that assemble to form the microfibrils and regulate growth factors or other transfer processes. Here, we obtained the 31,274 bp genomic DNA sequences of Oxfibrillin. The coding sequence length was 5784 bp encoding 1927 amino acids with a VWD domain, EGF/cb-EGF domains, a TR domain, and a transmembrane domain. Oxfibrillin was positioned within the subgroup of invertebrates and showed low scores for homology to mammalian fibrillin. In gene expression analysis, Oxfibrillin genes were constantly upregulated during the early regeneration process and then remained stable until the formation of the complete tail which indicated that it might be a vital factor to affect posterior regeneration process. Therefore, the Oxfibrillin of O. xiamen might play important roles in the regeneration process.

Buried treasure in a public repository: Mining mitochondrial genes of 32 annelid species from sequence reads deposited in the Sequence Read Archive (SRA).

Background: The mitochondrial genomes (mitogenomes) of metazoans generally include the same set of protein-coding genes, which ensures the homology of mitochondrial genes between species. The mitochondrial genes are often used as reference data for species identification based on genetic data (DNA barcoding). The need for such reference data has been increasing due to the application of environmental DNA (eDNA) analysis for environmental assessments. Recently, the number of publicly available sequence reads obtained with next-generation sequencing (NGS) has been increasing in the public database (the NCBI Sequence Read Archive, SRA). Such freely available NGS reads would be promising sources for assembling mitochondrial protein-coding genes (mPCGs) of organisms whose mitochondrial genes are not available in GenBank. The present study aimed to assemble annelid mPCGs from raw data deposited in the SRA. Methods: The recent progress in the classification of Annelida was briefly introduced. In the present study, the mPCGs of 32 annelid species of 19 families in clitellates and allies in Sedentaria (echiurans and polychaetes) were newly assembled from the reads deposited in the SRA. Assembly was performed with a recently published pipeline mitoRNA, which includes cycles of Bowtie2 mapping and Trinity assembly. Assembled mPCGs were deposited in GenBank as Third Party Data (TPA) data. A phylogenetic tree was reconstructed with maximum likelihood (ML) analysis, together with other mPCGs deposited in GenBank. Results and Discussion: mPCG assembly was largely successful except for Travisia forbesii; only four genes were detected from the assembled contigs of the species probably due to the reads targeting its parasite. Most genes were largely successfully obtained, whereas atp8, nad2, and nad4l were only successful in 22-24 species. The high nucleotide substitution rates of these genes might be relevant to the failure in the assembly although nad6, which showed a similarly high substitution rate, was successfully assembled. Although the phylogenetic positions of several lineages were not resolved in the present study, the phylogenetic relationships of some polychaetes and leeches that were not inferred by transcriptomes were well resolved probably due to a more dense taxon sampling than previous phylogenetic analyses based on transcriptomes. Although NGS data are generally better sources for resolving phylogenetic relationships of both higher and lower classifications, there are ensuring needs for specific loci of the mitochondrial genes for analyses that do not require high resolutions, such as DNA barcoding, eDNA, and phylogenetic analysis among lower taxa. Assembly from publicly available NGS reads would help design specific primers for the mitochondrial gene sequences of species, whose mitochondrial genes are hard to amplify by Sanger sequencing using universal primers.

Discovery and Genetic Characterization of a Vestimentiferan, Lamellibrachia satsuma, from the Submarine Volcano Omuro Dashi in the Izu-Ogasawara Arc.

Vestimentiferan tube worms (Annelida: Siboglinidae) were discovered in a hydrothermal field at a depth of 195 m in the crater of the submarine volcano Omuro Dashi in the Izu-Ogasawara Arc. Based on the nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene in individuals sampled in 2022, they were identified as Lamellibrachia satsuma Miura, Tsukahara & Hashimoto, 1997. STRUCTURE analysis and discriminant analysis of principal components (DAPC) based on 14 microsatellite markers showed a large genetic deviation of the population of Omuro Dashi from those of Kagoshima Bay and the north Mariana Arc (the Nikko and Daikoku Seamounts), whereas the population of Omuro Dashi did not show significant genetic deviation from that of the Nikko Seamount based on the COI gene. All analyses showed that individuals of a methane seep area on the Kanasu-No-Se Bank, the Nankai Trough, which were collected only in 1994, were more closely related to those of Omuro Dashi than to those of other habitats. These results suggest that the ancestors of the Omuro Dashi and Nankai Trough populations originated from migrants from the north Mariana Arc and that there might be undiscovered source population(s) of L. satsuma around the Nankai Trough.

Disjunct habitat of cryptic Terebellides (Annelida, Trichobranchidae) species shows a phylogenetic link between polychaetes from the White and the North Seas.

Understanding the distribution and biodiversity of marine species is crucial for developing effective conservation strategies and maintaining the health of global ecosystems. Advancements in molecular data utilization have significantly improved our understanding of biodiversity within the genus Terebellides. In this study, we conducted a phylogenetic analysis on polychaete samples from the Kandalaksha Bay, White Sea, revealing their affiliation with a putative undescribed species of the genus Terebellides found in two locations of the North Sea. Interestingly, this species was not detected in the Norwegian and Barents Seas, leading us to propose a disjunct distribution scenario for this Terebellides species. This unique distribution pattern might be attributed to the succession of polychaetes by new species, facilitated by the Gulf Stream and a climate change role in driving shifts in species' ranges and altering marine ecosystem dynamics.

Fast cycling culture of the annelid model Platynereis dumerilii.

Platynereis dumerilii, a marine annelid, is a model animal that has gained popularity in various fields such as developmental biology, biological rhythms, nervous system organization and physiology, behaviour, reproductive biology, and epigenetic regulation. The transparency of P. dumerilii tissues at all developmental stages makes it easy to perform live microscopic imaging of all cell types. In addition, the slow-evolving genome of P. dumerilii and its phylogenetic position as a representative of the vast branch of Lophotrochozoans add to its evolutionary significance. Although P. dumerilii is amenable to transgenesis and CRISPR-Cas9 knockouts, its relatively long and indefinite life cycle, as well as its semelparous reproduction have been hindrances to its adoption as a reverse genetics model. To overcome this limitation, an adapted culturing method has been developed allowing much faster life cycling, with median reproductive age at 13-14 weeks instead of 25-35 weeks using the traditional protocol. A low worm density in boxes and a strictly controlled feeding regime are important factors for the rapid growth and health of the worms. This culture method has several advantages, such as being much more compact, not requiring air bubbling or an artificial moonlight regime for synchronized sexual maturation and necessitating only limited water change. A full protocol for worm care and handling is provided.