Sea Anemone Species Research Articles

Diversity of biological activities of crude venom extracted from five species of South China Sea anemones

Developing novel, efficient, and safe peptide drugs from sea anemones has aroused great interest in countries around the world today. Sea anemones contain complex protein and peptide toxins, which determine the diversity of their biological activities. In this study, a variety of activities were assessed for crude venom extracted from five species of South China Sea anemones, including hemolytic, enzyme inhibition, anticancer, insecticidal, analgesic and lethal activities. The most toxic sea anemone was found to be Heteractis magnifica, which has high lethal activity in mice with an LD50 of 11.0 mg/kg. The crude venom of H. magnifica also exhibited a range of the most potent activities, including hemolytic, trypsin inhibitory, cytotoxic activity against U251 and A549 cells, insecticidal and analgesic activities. In addition, the crude venom of Stichodactyla haddoni was the most effective inhibitor of pepsin, and the crude venom of Heteractis crispa was extremely strong toxicity to HepG2 cells. These findings are of great significance for exploring the potential and application of South China Sea anemone resources, and are expected to provide new directions and possibilities for the development of novel anticancer drugs, analgesics and biopesticides.

Two new records of sea anemones (Cnidaria, Anthozoa, Actiniaria) from the Eastern Pacific coast of El Salvador

Two species of the genus Actinostella Duchassaing, 1850 are documented for the first time on the coasts of El Salvador: A. bradleyi (Verrill, 1869) and A. californica (McMurrich, 1893). Both species are briefly described, and images of live specimens showing their main external features are provided, as well as images of histological sections showing some taxonomic characteristics of their internal anatomy. These two species are distinguished from each other primarily by the number of tentacles, the number of mesentery pairs, the arrangement of gametogenic tissue, and some differences in their cnidom. The record of these two species in El Salvador increases the number of sea anemones recorded for the country to 12. Additionally, an updated list of sea anemone species documented in El Salvador is provided.

Molecular Insights into the Low Complexity Secreted Venom of Calliactis polypus.

Sea anemones are venomous animals that rely on their venom for prey capture, defense against predators, and intraspecific competition. Currently, comprehensive molecular and evolutionary analyses of the toxin repertoire for sea anemones are limited by a lack of proteomic data for most species. In this study, proteo-transcriptomic analysis was used to expand our knowledge of the proteinaceous components of sea anemone venom by determining the secreted venom proteome of Calliactis polypus. Electromechanical stimulation was used to obtain the secreted venom of C. polypus. We identified a low complexity proteome that was dominated by toxins with similarity to known neurotoxins, as well as six novel toxin candidates. The novel putative toxin candidates were found to be taxonomically restricted to species from the superfamily Metridioidea. Furthermore, the secreted venom of C. polypus had only three putative toxins in common with the venom of acontia from the same species and little similarity with the secreted venom of closely related species. Overall, this demonstrates that regionalized and lineage-specific variability in toxin abundance is common among sea anemone species. Moreover, the limited complexity of the toxin repertoire found in C. polypus supports the idea that peptide neurotoxins make up the dominant toxin arsenal found in the venom of sea anemones.

Social predation by a nudibranch mollusc.

Social predation is a common strategy used by predators to subdue and consume prey. Animals that use this strategy have many ways of finding each other, organizing behaviors and consuming prey. There is wide variation in the extent to which these behaviors are coordinated and the stability of individual roles. This study characterizes social predation by the nudibranch mollusc, Berghia stephanieae, which is a specialist predator that eats only the sea anemone, Exaiptasia diaphana. A combination of experimental and modeling approaches showed that B. stephanieae does predate upon E. diaphana in groups. The extent of social feeding was not altered by length of food deprivation, suggesting that animals are not shifting strategies based on internal state. It was unclear what cues the individual Berghia used to find each other; choice assays testing whether they followed slime trails, were attracted to injured anemones, or preferred conspecifics feeding did not reveal any cues. Individuals did not exhibit stable roles, such as leader or follower, rather the population exhibited fission-fusion dynamics with temporary roles during predation. Thus, the Berghia provides an example of a specialist predator of dangerous prey that loosely organizes social feeding, which persists across hunger states and uses temporary individual roles; however, the cues that it uses for aggregation are unknown.

Regulation of Green Fluorescent Proteins by Sea Anemones (Anthopleura spp.) in Response to Light.

Green Fluorescent Proteins (GFPs) are a family of proteins with a disjunct systematic distribution; their biological functions remain speculative for the most part. Here we report studies of 3 closely related species of green sea anemones (Anthopleura) that express GFPs throughout their ectoderm. Individuals of these species maintain facultative symbiosis with zooxanthellae in their endoderm and inhabit the rocky intertidal or shallow subtidal. Thus, they depend on exposure to light to maintain photosynthesis of their symbionts, and simultaneously need to manage stresses associated with this exposure. We present experimental evidence that these sea anemones regulate the amount of GFP in their bodies in response to the surrounding light environment: they increase or reduce GFP when exposed to brighter or dimmer light, respectively, yet they maintain some GFP while in darkness, for surprisingly long periods.

Towards the Exploration and Evolution of Insulin-like Venoms in Actiniaria (Sea anemones).

Recent studies have elucidated the diversity of genes encoding venom in Sea anemones. However, most of those genes are yet to be explored in an evolutionary context. Insulin is a common peptide across metazoans and has been coopted into a predatory venom in many venomous lineages. In this study, we focus on the diversity of insulin-derived venoms in Sea anemones and on elucidating their evolutionary history. We sourced data for 34 species of Sea anemones and found sequences belonging to two venom families which have Insulin PFAM annotations. Our findings show that both families have undergone duplication events. Members of each of the independently evolving clades have consistent predicted protein structures and distinct dN/dS values. Our work also shows that sequences allied with VP302 are part of a multidomain venom contig and have experienced a secondary gain into the venom system of cuticulate Sea anemones.

Evolutionary Analysis of Cnidaria Small Cysteine-Rich Proteins (SCRiPs), an Enigmatic Neurotoxin Family from Stony Corals and Sea Anemones (Anthozoa: Hexacorallia).

Cnidarians (corals, sea anemones, and jellyfish) produce toxins that play central roles in key ecological processes, including predation, defense, and competition, being the oldest extant venomous animal lineage. Cnidaria small cysteine-rich proteins (SCRiPs) were the first family of neurotoxins detected in stony corals, one of the ocean's most crucial foundation species. Yet, their molecular evolution remains poorly understood. Moreover, the lack of a clear classification system has hindered the establishment of an accurate and phylogenetically informed nomenclature. In this study, we extensively surveyed 117 genomes and 103 transcriptomes of cnidarians to identify orthologous SCRiP gene sequences. We annotated a total of 168 novel putative SCRiPs from over 36 species of stony corals and 12 species of sea anemones. Phylogenetic reconstruction identified four distinct SCRiP subfamilies, according to strict discrimination criteria based on well-supported monophyly with a high percentage of nucleotide and amino acids' identity. Although there is a high prevalence of purifying selection for most SCRiP subfamilies, with few positively selected sites detected, a subset of Acroporidae sequences is influenced by diversifying positive selection, suggesting potential neofunctionalizations related to the fine-tuning of toxin potency. We propose a new nomenclature classification system relying on the phylogenetic distribution and evolution of SCRiPs across Anthozoa, which will further assist future proteomic and functional research efforts.

First Record of Sea Anemone Anthopleura handi Dunn, 1978 (Actiniaria: Actinidae) from Coast across the Indian Mainland

Anthopleura handi Dunn, 1978 was previously recorded from Andaman Islands. Here we report Anthopleura handi Dunn, 1978 for the first time from coast across the Indian mainland. The sea anemone specimen was collected from intertidal sandy area of Talasari Beach (21°36'11"N, 87°24'27"E), Balasore, Odisha. This paper also reports the species for the first time from eastern coast of Indian Mainland as well as from Odisha state with its diagnostic characters. In India there are total thirteen species of genus Anthopleura have been reported without proper taxonomic study. Thus, this paper also deals with a comparative analysis of various sea anemone species of genus Anthopleura reported from Indian water.

Isolation and cDNA cloning of four peptide toxins from the sea anemone Heteractis aurora.

Sea anemones are well known to contain multiple peptide toxins. However, of more than 1100 species of sea anemones distributed worldwide, only a little over 50 have been studied for peptide toxins. Therefore, innumerable unique and novel peptide toxins remain to be discovered in unstudied sea anemones. Isolation of peptide toxins in the sea anemone Heteractis aurora was attempted by gel filtration and reverse-phase high performance liquid chromatography, using the toxicity to crabs as an index. The amino acid sequences of the isolated four toxins (Hau I-IV) and their precursors were determined using a combination of protein sequencing and cDNA cloning. Hau I and IV were potently lethal to crabs, whereas Hau II and III were only paralytic. The precursor proteins of the four toxins were commonly composed of a signal peptide, a propart, and the remaining region including a mature peptide. Interestingly, four and two copies of the mature peptide were present in the precursor proteins of Hau II and III, respectively. Homology searches revealed that Hau I (30 amino acid residues) is a novel peptide toxin, although it has the same cysteine pattern CXXC-C-C as the boundless β-hairpin (BBH) family. Hau II (27 amino acid residues) and III (28 amino acid residues) were homologous with the BBH family, whereas Hau IV (49 amino acid residues) was a new member of the well-known type 1 sodium channel toxin family. This study showed that a novel class of toxin (Hau I), two BBH family toxins (Hau II and III), and a type 1 sodium channel toxin (Hau IV) are present in the toxin of the sea anemone H. aurora.

Notes on the morphology, distribution and bionomics of Phyllodiscus semoni (Anthozoa: Actiniaria: Aliciidae)

Phyllodiscus semoni Kwietniewski, 1897, a little-known species of sea anemones of the family Aliciidae, is quite widely distributed in the tropics but has not been recorded from the Spratly Islands so far. The reason that the range of this species is poorly studied lies not only in the fact that many tropical areas are still poorly explored but also in the peculiar appearance of this animal in the sea. It rarely extends its tentacles during daylight hours and is not readily recognised by divers and researchers as an actinian. Our underwater observations, accompanied by high-resolution photography and specimen collection, evidence the presence of this species along the continental coast of Vietnam and in the waters of the Spratly Archipelago. During the dives, some observations were made on the behavior of the polyps and symbiotic shrimp living on their bodies. This species of sea anemones can cause severe skin burns in humans, which is confirmed by our experience. Summarised literature data on the distribution of Ph. semoni is provided.

Mitochondrial and nuclear gene sequencing confirms the presence of the invasive sea anemone Diadumene lineata (Verrill, 1869) (Cnidaria: Actiniaria) in Argentina.

Diadumene lineata is one of the most widespread sea anemone species worldwide. Although this species has been reported a few times on the Argentine coast since 2004, its identification has traditionally been based on external morphological characteristics, and in most cases no voucher specimens are available to support previous records. In this study, we obtained DNA sequences of two mitochondrial markers (12S and 16S) and two nuclear markers (18S and 28S) from specimens of D. lineata collected in two locations on the Argentine coast separated by almost 800 km. Additionally, we conducted an analysis of the morphology, as well as the types and size ranges of cnidae, using specimens collected at three different locations along the Argentine coast. Furthermore, since introduced populations of D. lineata are presumably ephemeral and only reproduce asexually outside their native range, we examined the internal anatomy of representatives from the Argentine coast for gametogenic tissue as an indication of whether they might be capable of sexual reproduction. DNA data support our morphological identification, including cnidae analyses, of the specimens as D. lineata. Furthermore, all specimens examined were determined to be sterile. Genetic sequence comparisons, phylogenetic reconstruction, and cnidae data support the identification of individuals of D. lineata from Mar Chiquita and Garipe Beach, confirming the presence of the species on the Argentine coast using both morphological and molecular tools. The absence of fertile specimens suggests that each sampled population is likely reproducing only by asexual reproduction and possibly composed of clones. The presence of an additional category of longer p-mastigophores B2a in the actinopharynx and filaments, as well as holotrichs in the column, is also reported. For the first time, we have confirmed the presence of D. lineata in Argentina through molecular data. Additionally, our findings indicate that the analyzed specimens are sterile, suggesting that this species is not engaging in sexual reproduction in the studied localities. It is crucial to continue monitoring the populations of D. lineata along the Argentine coast to assess whether they establish sexual reproduction, expand their distribution range or disappear, or potentially cause any harm to local species or alterations in benthic communities.

Bellactis lux n. sp. (Cnidaria: Anthozoa: Actiniaria: Aiptasiidae), a new sea anemone from the Gulf of Mexico.

Here we describe a new species of sea anemone from the family Aiptasiidae based on specimens collected from the Gulf of Mexico (USA: Florida & Alabama). Accounts of this species have been known since the early 1990s, primarily from an underwater field guide and hobbyist aquarium literature under the name Lightbulb Anemone. We describe it as a new species from the genus Bellactis based on anatomy, histology, and cnidom. Members of this species are small in size, with a smooth, typically contracted column divided into regions based on color and bearing rows of two or three elevated cinclides in the mid column. Their tentacles are distinctive, translucent, distally inflated and can be bulbous in shape, with sub annular rings. This description synthesizes information about Bellactis and contextualizes what is known about its diversity in light of other members of the Aiptasiidae.

Litophellia enoplosa sp. nov., a new species of sea anemone (Actiniaria, Metridioidea) from cold-water coral environments in the North-western Mediterranean Sea

A new species of sea anemone (Actiniaria, Metridioidea) is described and illustrated from cold-water coral environments at 600 m depth at the Blanes Canyon in the north-western Mediterranean Sea. Litophellia enoplosa sp. nov. is characterized by a column divisible into scapus and scapulus, the former with cuticle forming tenaculi, a mesogleal marginal sphincter muscle, more mesenteries proximally than distally arranged octomerously, and a single pair of directive mesenteries; it has strong retractor and parietobasilar muscles, and acontia with basitrichs and p-mastigophores B2a. Litophellia enoplosa sp. nov. is the second species described in the genus and it can be distinguished from the other species by the cuticle in the column, size ranges of the cnidae, and geographic and bathymetric distribution.

Establishment of a New Model Sea Anemone for Comparative Studies on Cnidarian-Algal Symbiosis.

Frequent coral bleaching has drawn attention to the mechanisms of coral dinoflagellate endosymbiosis. Owing to the difficulty of rearing corals in the laboratory, model symbiosis systems are desired. The sea anemone Exaiptasia diaphana, hosting clade B1 of the genus Breviolum, has long been studied as a model system; however, a single species is insufficient for comparative studies and thus provides only limited resources for symbiosis research, especially regarding the specificity of host-symbiont associations. We established a clonal strain of the sea anemone Anthopleura atodai, whose symbiont was identified as a novel subclade of Symbiodinium (clade A) using a novel feeding method. We also developed a method to efficiently bleach various sea anemone species using a quinoclamine-based herbicide. Bleached A. atodai polyps were vital and able to reproduce asexually, exhibiting no signs of harmful effects of the drug treatment. Pilot studies have suggested that host-symbiont specificity is influenced by multiple steps differently in A. atodai and E. diaphana. RNAseq analyses of A. atodai showed that multiple NPC2 genes were expressed in the symbiotic state, which have been suggested to function in the transport of sterols from symbionts to host cells. These results reveal the usefulness of A. atodai in comparative studies of cnidarian-algal symbiosis.

Multiomic Approach for Bioprospection: Investigation of Toxins and Peptides of Brazilian Sea Anemone Bunodosoma caissarum.

Sea anemones are sessile invertebrates of the phylum Cnidaria and their survival and evolutive success are highly related to the ability to produce and quickly inoculate venom, with the presence of potent toxins. In this study, a multi-omics approach was applied to characterize the protein composition of the tentacles and mucus of Bunodosoma caissarum, a species of sea anemone from the Brazilian coast. The tentacles transcriptome resulted in 23,444 annotated genes, of which 1% showed similarity with toxins or proteins related to toxin activity. In the proteome analysis, 430 polypeptides were consistently identified: 316 of them were more abundant in the tentacles while 114 were enriched in the mucus. Tentacle proteins were mostly enzymes, followed by DNA- and RNA-associated proteins, while in the mucus most proteins were toxins. In addition, peptidomics allowed the identification of large and small fragments of mature toxins, neuropeptides, and intracellular peptides. In conclusion, integrated omics identified previously unknown or uncharacterized genes in addition to 23 toxin-like proteins of therapeutic potential, improving the understanding of tentacle and mucus composition of sea anemones.

Cloning and structural basis of fluorescent protein color variants from identical species of sea anemone, Diadumene lineata

Diadumene lineata is a colorful sea anemone with orange stripe tissue of the body column and plain tentacles with red lines. We subjected Diadumene lineata to expression cloning and obtained genes encoding orange (OFP: DiLiFP561) and red fluorescent proteins (RFPs: DiLiFP570 and DiLiFP571). These proteins formed obligatory tetramers. All three proteins showed bright fluorescence with the brightness of 58.3 mM−1·cm−1 (DiLiFP561), 43.9 mM−1·cm−1 (DiLiFP570), and 31.2 mM−1·cm−1 (DiLiFP571), which were equivalent to that of commonly used red fluorescent proteins. Amplitude-weighted average fluorescence lifetimes of DiLiFP561, DiLiFP570 and DiLiFP571 were determined as 3.7, 3.6 and 3.0 ns. We determined a crystal structure of DiLiFP570 at 1.63 Å resolution. The crystal structure of DiLiFP570 revealed that the chromophore has an extended π-conjugated structure similar to that of DsRed. Most of the amino acid residues surrounding the chromophore were common between DiLiFP570 and DiLiFP561, except M159 of DiLiFP570 (Lysine in DiLiFP561), which is located close to the chromophore hydroxyl group. Interestingly, a similar K-to-M substitution has been reported in a red-shifted variant of DsRed (mRFP1). It is a striking observation that the naturally evolved color-change variants are consistent with the mutation induced via protein engineering processes. The newly cloned proteins are promising as orange and red fluorescent markers for imaging with long fluorescence lifetime.Graphical abstract

Single-cell atavism reveals an ancient mechanism of cell type diversification in a sea anemone

Cnidocytes are the explosive stinging cells unique to cnidarians (corals, jellyfish, etc). Specialized for prey capture and defense, cnidocytes comprise a group of over 30 morphologically and functionally distinct cell types. These unusual cells are iconic examples of biological novelty but the developmental mechanisms driving diversity of the stinging apparatus are poorly characterized, making it challenging to understand the evolutionary history of stinging cells. Using CRISPR/Cas9-mediated genome editing in the sea anemone Nematostella vectensis, we show that a single transcription factor (NvSox2) acts as a binary switch between two alternative stinging cell fates. Knockout of NvSox2 causes a transformation of piercing cells into ensnaring cells, which are common in other species of sea anemone but appear to have been silenced in N. vectensis. These results reveal an unusual case of single-cell atavism and expand our understanding of the diversification of cell type identity.

First In Situ Observation of Sperm Release in Corynactis carnea (Anthozoa: Corallimorpharia) from Patagonia, Argentina

Jewel sea anemones constitute a relatively small group of solitary cnidarians, a sister group of scleractinian corals. In the southwest Atlantic Ocean off Argentina, two species of jewel sea anemones have been found: Corynactis carnea and Corallimorphus rigidus. Corynactis carnea is a common and abundant species in shallow water of northern Atlantic Patagonia, but reproductive data on this species is scarce; the species is known to reproduce asexually. During a SCUBA diving survey in an Atlantic rocky reefs (20 m depth) in Patagonia, we observed for the first time specimens of C. carnea releasing sperm, eight days after the full moon and during the summer season while spawning has been previously observed in at least three species of Corynactis, but all were recorded as occurring in winter.

First record of the epiphyte sea anemone Bunodeopsis pelagica in the Mexican Atlantic

Abstract Bunodeopsis pelagica is one of the few species of sea anemones known to be found in the floating algae of Sargassum spp. It has been reported in the North Atlantic, particularly in the Canary Islands and the coast of Jamaica. The present study is the first to report the occurrence of B. pelagica in the Mexican Atlantic, found as epiphyte on Sargassum natans off Alacranes reef (southern Gulf of Mexico) and in Puerto Morelos reef (Mexican Caribbean), revealing that this species is much more widespread than has been reported. Bunodeopsis pelagica is mainly distinguished from other species of Bunodeopsis by having simple minute protuberances on its column, while its congeners have larger and much more developed vesicular projections. With the record of B. pelagica , the number of known species of actiniarian sea anemones from the Mexican Atlantic increases to 23.

Sea anemone microhabitats enhance the diversity and biomass of mobile invertebrates on temperate rocky shores

Sea anemones of the genus Anthopleura are abundant in rocky intertidal communities of the eastern Pacific, but their role as habitat and facilitators of biodiversity in this system has been overlooked. We provide evidence for a previously undescribed mechanism of facilitation associated with these cnidarians on temperate rocky shores. Sea anemones in the genus Anthopleura slowly release stored water at low tide, which helps them maintain ideal temperature conditions via evaporative cooling. This behavior results in leakage of water to surrounding intertidal habitats, producing unique microhabitats in the upper intertidal zone. We hypothesized that mobile invertebrate communities would use this habitat during low tides to ameliorate abiotic stress. In an in situ experiment spanning 3 sites along the coast of California, USA, it was found that maximum temperatures and relative desiccation rates were lower in sea anemone-associated microhabitats as compared to adjacent rock habitats. Observational surveys revealed higher richness and biomass of mobile invertebrates in sea anemone microhabitats, and these differences were supported by manipulative experiments that included both adding and removing sea anemones. Our results support a novel community-wide facilitation driven by an adaptation of intertidal sea anemones. The effects of sea anemone microhabitat on upper shore biodiversity were associated with site and sea anemone species and may suggest a facilitative role of a temperate cnidarian-algal mutualism in mitigating stress and enhancing biodiversity.