Sipuncula Research Articles

Unusual polyunsaturated fatty acids in edible marine worms identified by covalent adduct chemical ionization mass spectrometry

Non-traditional seafood, such as spoon worms (Urechis unicinctus) and peanut worms (Sipunculus nudus), serves as both delicacies and potential solutions to the global food insecurity crisis. Despite being consumed primarily in parts of China, Korea, and Japan, the nutritional values especially the complex fatty acid compositions of these marine worms are difficult to characterize. To overcome this obstacle, we employed covalent adduct chemical ionization (CACI) tandem mass spectrometry for the de novo identification of their unusual polyunsaturated fatty acids (PUFA). Through this method, we identified several PUFA with polymethylene-interrupted (PMI) double bond configurations, including 22:3(7Z,13Z,16Z), a novel PUFA derived from sciadonic acid. U. unicinctus exhibits an exceptionally low n-6/n-3 PUFA ratio of 0.15, making it a potential functional food to counterbalance the n-6/n-3 imbalance in modern diets. S. nudus boasts notably high concentrations (∼3 %, wt/wt) of branched chain fatty acids (BCFA), exceeding typical levels found in dairy products.

Phascolion kamchatikus sp. nov., the first interstitial sipunculan (Sipuncula, Phascolionidae) from the Pacific Ocean

A new species of meiobenthic sipunculan, Phascolion kamchatikus sp. nov. collected from coarse sand and shell hash in shallow waters in the northwestern Pacific, is described and illustrated using light and electron microscopy. This new species is well distinguished from all other congeners by a unique combination of main taxonomic characters. This is the only sipunculan having only two primary tentacles in both juvenile and sexually mature worms. There are no hooks and holdfast papillae in the adult stages. Because of the presence of three nearly equal in size retractor muscles forming a retractor column divided for most of its length, this species is assigned to the subgenus Isomya of the genus Phascolion. Unlike most other species of the genus Phascolion, P. kamchatikus sp. nov. has an interstitial mode of life moving among the coarse sand grains and shell hash accumulated between rocks and stones in shallow water areas alongside the Pacific coast of Kamchatka, Aleutian Islands and Alaska Peninsula. Phascolion kamchatikus sp. nov. constitutes the only third interstitial species of the phylum Sipuncula. This is also the ninth species of Phascolion in the northwestern Pacific and the first interstitial representative of this genus in the Pacific Ocean.

HIGHLY BIOERODED RHODOLITHS FROM THE VITÓRIA-TRINDADE SEAMOUNT CHAIN (BRAZIL, SOUTHWESTERN ATLANTIC)

Abstract Rhodoliths are nodules mainly composed of crustose coralline algae with subordinate encrusting organisms, formed by successive overlapping encrustation. The subspheroidal rhodoliths from the Vitória-Trindade Seamount Chain (Jaseur Seamount and Trindade Island shelf; Brazil, southwestern Atlantic), sampled at water depths from 65 to 74 m, were built by crustose coralline algae (Harveylithon, Lithophyllum, Lithothamnion, Mesophyllum, Roseolithon, and Sporolithon) and subordinated encrusting foraminifera (agglutinated, unidentified hyaline, and Homotrema rubrum), bryozoans, serpulids, and balanids. Successive taphonomic phases of bioerosion, boring filling, and cement precipitation modified the original rhodolith inner structures resulting in a complex structureless mass of coralline algal fragments, encrusting organisms, borings, lithified fine-grained sediment, and carbonate cement. Borings include the ichnogenera Entobia (produced by etching sponges), Gastrochaenolites (boring bivalves) and Trypanites (polychaetes and sipunculid worms). The lithification of the material filling the borings (micrite and carbonate cements) created new substrates for subsequent bioerosion. Radiocarbon dating of selectively sampled invertebrate and algal skeletons in the rhodolith interior yielded calibrated ages of hundreds of years (up to 912 ± 152 years cal. BP on the Trindade insular shelf and up to 763 ± 131 years cal. BP on Jaseur Seamount). These values indicate growth rates from 0.1 to 0.5 mm/year, which are significantly higher than those recorded from rhodoliths at similar depths on the Brazilian shelf. Low sedimentation settings and high productivity at the tops of the seamounts and island shelf promoted the growth of nodule frame builders, both autotrophs and heterotrophs, and favored continuous activity of bioeroders.

Peanut worms as unreported fishery resources in Brazil

Peanut worms as unreported fishery resources in Brazil

Influence of sipunculan (peanut worm) activity on orifice formation in scleractinian Heterocyathus for adaptation to soft substrates

Mutualism profoundly affects the morphology and ecological evolution of both hosts and symbionts involved. Heterocyathus is a solitary scleractinian coral that lives on soft substrata, and sipunculan worms live symbiotically in the tube-like cavities (orifice) inside the coral skeletons. This habitat provides protection to the sipunculan worms against predators and—owing to the mobility of the worms—prevents the coral from being buried with sediments. The orifice growth is closely related to the symbiont sipunculan worms; however, this has not been previously elucidated. Here, we clarified the growth process of scleractinian coral orifices and the influence of sipunculan activity on this. The orifices were originally formed by rapid accretion deposits. The coral soft tissue enveloping the growth edge of the orifice repeatedly retreated to the outer side due to direct damage to the soft part and/or excessive stress caused by the rubbing of the sipunculan through locomotion, excretion, and feeding behaviour. This resulted in a toppled-domino microskeletal structure appearance and maintenance of the orifice growth. These outcomes demonstrate the first example of the direct influence of symbionts on the skeletal morphogenesis of scleractinian corals. The mutualism between the two organisms is maintained by the beneficial confrontation in forming orifices.

Rapid Separation and Display of Active Fibrinogenolytic Agents in Sipunculus nudus through Fibrinogen-Polyacrylamide Gel Electrophoresis.

Fibrinogenolytic agents that can dissolve fibrinogen directly have been widely used in anti-coagulation treatment. Generally, identifying new fibrinogenolytic agents requires the separation of each component first and then checking their fibrinogenolytic activities. Currently, polyacrylamide gel electrophoresis (PAGE) and chromatography are mostly used in the separating stage. Meanwhile, the fibrinogen plate assay and reaction products based PAGE are usually adopted to display their fibrinogenolytic activities. However, because of the spatiotemporal separation of those two stages, it is impossible to separate and display the active fibrinogenolytic agents with the same gel. To simplify the separating and displaying processes of fibrinogenolytic agent identification, we constructed a new fibrinogen-PAGE method to rapidly separate and display the fibrinogenolytic agents of peanut worms (Sipunculus nudus)in this study. This method includes fibrinogen-PAGE preparation, electrophoresis, renaturation, incubation, staining, and decolorization. The fibrinogenolytic activity and molecular weight of the protein can be detected simultaneously. According to this method, we successfully detected more than one active fibrinogenolytic agent of peanut wormhomogenate within 6 h. Moreover, this fibrinogen-PAGE method is time and cost-friendly. Furthermore, this method could be used to study the fibrinogenolytic agents of the other organisms.

Sipuncula (Peanut Worms) in Indonesia Waters: A Review

Sipunculans, or peanut worms, are unsegmented worm-like marine organisms with distinctive trunk-like bodies. They play important roles in marine ecosystems, such as bioerosion, bioturbation, and food sources. Sipunculans are also harvested as human food in some regions worldwide. Despite their importance, a comprehensive understanding of the Sipuncula in Indonesia is still limited. This work aimed to compile a study of the distribution, ecology, and potency of Sipuncula species in Indonesian waters. The distribution of Sipuncula species in Indonesian waters was recorded from the northernmost to the easternmost part of Indonesia. So far, nineteen species have been found in Indonesian waters and are dominated by Sipunculus nudus. In Indonesia, Sipuncula exhibits a diverse range of habitats, they can be found in seagrass, mangrove and coral reef ecosystems, in both sandy and muddy sediments. Sipunculans found in Indonesia have ecosystem function as bioturbator, shell-utilizer, and coral-symbiotic species. Sipuncula also has potency as a food and nutraceutical for human health maintenance because it contains highly nutritious such as protein, carbohydrates, ash, lipids, moisture, minerals, amino acids, and fatty acids. In some areas in Indonesia, local communities use Sipuncula as a traditional food source and fishing bait. Given its significance for marine ecosystems and human livelihoods in Indonesia, future management strategies should include regulations on catch size and management practices, data collection, promotion of sustainable fishing practices, and research on Sipuncula biology and ecology. Addressing these knowledge gaps will help to ensure the sustainable use and conservation of Sipuncula in Indonesia.

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.

Novel cytoskeletal traits in the intestinal parasites (Squirmida, Platyproteum vivax) of Pacific peanut worms (Sipuncula, Phascolosoma agassizii).

The cytoskeletal organization of a squirmid, namely Platyproteum vivax, was investigated with confocal laser scanning microscopy (CLSM) to refine inferences about convergent evolution among intestinal parasites of marine invertebrates. Platyproteum inhabits Pacific peanut worms (Phascolosoma agassizii) and has traits that are similar to other lineages of myzozoan parasites, namely gregarine apicomplexans within Selenidium, such as conspicuous feeding stages, called "trophozoites," capable of dynamic undulations. SEM and CLSM of P. vivax revealed an inconspicuous flagellar apparatus and a uniform array of longitudinal microtubules organized in bundles (LMBs). Extreme flattening of the trophozoites and a consistently oblique morphology of the anterior end provided a reliable way to distinguish dorsal and ventral surfaces. CLSM revealed a novel system of microtubules oriented in the flattened dorsoventral plane. Most of these dorsoventral microtubule bundles (DVMBs) had a punctate distribution and were evenly spaced along a curved line spanning the longitudinal axis of the trophozoites. This configuration of microtubules is inferred to function in maintaining the flattened shape of the trophozoites and facilitate dynamic undulations. The novel traits in Platyproteum are consistent with phylogenomic data showing that this lineage is only distantly related to Selenidium and other marine gregarine apicomplexans with dynamic intestinal trophozoites.

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.

Variasi Spasial Kepadatan Cacing Kacang di Pantai Toronipa Sulawesi Tenggara

The density of peanut worms is concentrated in the subtidal zone and is closely related to the substrate where they live. This research aims to determine spatial variations in peanut worm density. This research was carried out at Toronipa Beach, Southeast Sulawesi from June 2020 to February 2021. Peanut worms were collected using a hand scoop to a depth of ±10 cm in a 10x10 m2 quadrat transect at each station. Data were analyzed using standard formulas and tested using Mann-Whitney and linear regression. The results showed that the highest density of peanut worms was found at station II with a mean value of 12±104 ind/10m2 which was significantly different from stations I and III with densities of 9±52 ind/10m2 and 7±124 ind/10m2 respectively. Five types of seagrass were identified, namely Cymodocea rotundata, Enhalus acoroides, Halodule uninervis, Halophila minor, and Thalassia hemprichii. The number of seagrass species found at stations I, II, and III were 1, 2, and 5, respectively. Peanut worm density was positively correlated with seagrass density with a coefficient of determination of 86.45%. Water quality tends to be the same at all stations, except that organic matter is higher than at stations I and II. The dominant fine sand fraction was found at stations I and III, while very coarse sand was dominantly found at station II. The density of peanut worms on Toronipa Beach is determined by a combination of seagrass density, high levels of organic matter, and the dominant texture of the coarse sand substrate.
 
 Keywords: peanut worm, spatial density, Sulawesi, water quality

Characterization of acid-soluble collagen from peanut worm (Siphonosoma australe) and its inhibitory activity of cyclooxygenase-2

The aim of this study was to determine the effect of acetic acid extraction time on the physicochemical characteristics of peanut worm collagen and its ability to inhibit cyclooxygenase-2 (COX-2). The peanut worms were subjected to extraction using 0.5 M acetic acid for varying durations, followed by analysis for various physicochemical properties, including yield, protein content, pH, viscosity, molecular weight, amino acid composition, FTIR, and COX-2 inhibitory activity. The results showed that 72 h of extraction time was optimal for collagen production. The peanut worm collagen yields, protein, and pH were 3.55 ± 0.20%, 85.58 ± 0.10%, and 3.61, respectively. The relative viscosity of collagen decreased with increased temperature by 51.35%. Collagen consists of α and β chains with high levels of amino acids such as glutamic acid, arginine, and aspartic acid. SDS-PAGE analysis revealed that peanut worm collagen contained type I collagen. FTIR confirmed the collagen’s triple-helical structure. The COX-2 inhibitor assay showed that collagen inhibited the COX-2 enzyme by 41.12% ± 0.58% (IC50 = 59.9 μg mL–1). Therefore, this peanut worm collagen could serve as a potential substitute for terrestrial mammalian collagen as an ingredient in food, cosmetics, and other industries. In addition, peanut worm collagen can be used as a nutraceutical and medicine to prevent and treat inflammatory diseases.

Macronutrient Content of Sipunculus nudus as an Antistunting Candidate

Consuming balanced nutritious food with macronutrient content including carbohydrates, protein and fat for pregnant and breastfeeding mothers as well as children under five during the growth and development period is one of the efforts to prevent and treat anti-stunting, up to the age of 2 years. Waters and beaches are one of the habitats for marine biota such as bivalve shells, gastropods, various fish and other marine biota whose names and uses are not yet popular but are widely consumed by coastal communities, including Sipunculus nudus. Marine biota from the phylum Sipuncula in some areas are called sea worms, are one of the marine biota whose habitat is in the sand, and are the favorite food of coastal communities. Information about the nutritional and mineral content of this marine biota species has not been widely reported. This research aims to determine the macronutrient content of Sipunculus nudus collected from the coastal waters of Samboang, South Sulawesi. Determination of macronutrients was carried out using several methods, including for protein using the Kjeldahl method, fat using extraction with n-hexane solvent, water and ash content using an oven and furnace. The results of measuring the macronutrient content of Sipunculus nudus showed a water content of 72%, ash content of 1.19%, protein content of 6.5%, fat content of 2.07% and carbohydrate content using difference calculations was obtained at 18.24%.

Anti-Inflammatory Effects of Bioactive Compounds from Seaweeds, Bryozoans, Jellyfish, Shellfish and Peanut Worms.

Inflammation is a defense mechanism of the body in response to harmful stimuli such as pathogens, damaged cells, toxic compounds or radiation. However, chronic inflammation plays an important role in the pathogenesis of a variety of diseases. Multiple anti-inflammatory drugs are currently available for the treatment of inflammation, but all exhibit less efficacy. This drives the search for new anti-inflammatory compounds focusing on natural resources. Marine organisms produce a broad spectrum of bioactive compounds with anti-inflammatory activities. Several are considered as lead compounds for development into drugs. Anti-inflammatory compounds have been extracted from algae, corals, seaweeds and other marine organisms. We previously reviewed anti-inflammatory compounds, as well as crude extracts isolated from echinoderms such as sea cucumbers, sea urchins and starfish. In the present review, we evaluate the anti-inflammatory effects of compounds from other marine organisms, including macroalgae (seaweeds), marine angiosperms (seagrasses), medusozoa (jellyfish), bryozoans (moss animals), mollusks (shellfish) and peanut worms. We also present a review of the molecular mechanisms of the anti-inflammatory activity of these compounds. Our objective in this review is to provide an overview of the current state of research on anti-inflammatory compounds from marine sources and the prospects for their translation into novel anti-inflammatory drugs.

Transcriptome Responses to Different Environments in Intertidal Zones in the Peanut Worm Sipunculus nudus.

The peanut worm (Sipunculus nudus) is an important intertidal species worldwide. Species living in the same aquaculture area might suffer different environmental impacts. To increase knowledge of the molecular mechanisms underlying the response to environmental fluctuations, we performed a transcriptome analysis of S. nudus from different intertidal zones using a combination of the SMRT platform and the Illumina sequencing platform. (1) A total of 105,259 unigenes were assembled, and 23,063 unigenes were perfectly annotated. The results of the PacBio Iso-Seq and IIIumina RNA-Seq enriched the genetic database of S. nudus. (2) A total of 830 DEGs were detected in S. nudus from the different groups. In particular, 33 DEGs had differential expression in the top nine KEGG pathways related to pathogens, protein synthesis, and cellular immune response and signaling. The results indicate that S. nudus from different zones experience different environmental stresses. (3) Several DEGs (HSPA1, NFKBIA, eEF1A, etc.) in pathways related to pathogens (influenza A, legionellosis, measles, and toxoplasmosis) had higher expression in groups M and L. HSPA1 was clearly enriched in most of the pathways, followed by NFKBIA. The results show that the peanut worms from the M and L tidal flats might have suffered more severe environmental conditions. (4) Some DEGs (MKP, MRAS, and HSPB1) were upregulated in peanut worms from the H tidal flat, and these DEGs were mainly involved in the MAPK signaling pathway. These results indicate that the MAPK pathway may play a vital role in the immune response of the peanut worm to the effects of different intertidal flats. This study provides a valuable starting point for further studies to elucidate the molecular basis of the response to different environmental stresses in S. nudus.

The first high-quality chromosome-level genome of the Sipuncula Sipunculus nudus using HiFi and Hi-C data

Sipuncula is a class of exocoelomic unsegmented animals whose evolutionary relationships are unresolved. The peanut worm Sipunculus nudus is a globally distributed, economically important species belonging to the class Sipuncula. Herein, we present the first high-quality chromosome-level assembly of S. nudus based on HiFi reads and high-resolution chromosome conformation capture (Hi-C) data. The assembled genome was 1,427 Mb, with a contig N50 length of 29.46 Mb and scaffold N50 length of 80.87 Mb. Approximately 97.91% of the genome sequence was anchored to 17 chromosomes. A BUSCO assessment showed that 97.7% of the expectedly conserved genes were present in the genome assembly. The genome was composed of 47.91% repetitive sequences, and 28,749 protein-coding genes were predicted. A phylogenetic tree demonstrated that Sipuncula belongs to Annelida and diverged from the common ancestor of Polychaeta. The high-quality chromosome-level genome of S. nudus will serve as a valuable reference for studies of the genetic diversity and evolution of Lophotrochozoa.

Symbiont-Bearing Colonial Corals and Gastropods: An Odd Couple of the Shallow Seas

In order to investigate the serendipitous find of a gastropod encrusted by the symbiont-bearing colonial coral Oculina patagonica, we examined several specimens of cnidarian-encrusted gastropods, ranging in age from the Pliocene to the Recent, and characterized in detail their sclerobiont cover. The results of our analysis suggest that gastropod shells can be encrusted by symbiont-bearing colonial corals at various times: (1) when the gastropod is alive; (2) when the shell is being used by a secondary inhabitant (e.g., hermit crabs or sipunculid worms); (3) when the shell is discarded but yet to be buried. The relationship between the symbiont-bearing coral and the inhabitant(s) of the encrusted shell is an example of facultative mutualism, i.e., it is non-obligate yet beneficial for both ends as the former obtains the capability to move, and the latter improves the resistance and resilience of its armor, thus obtaining extra protection from predators. Being able to move could prove particularly useful for a symbiont-bearing coral because, in addition to removing the risk of being smothered by sediment, it would also favor the photosynthetic activity of its algal endosymbionts by allowing the coral to be always clean of sedimentary particles. Although the resulting epibiotic association would be limited in size by the ability of either the gastropod or the secondary inhabitant of the shell to move at the seafloor, these small and easy-to-miss benthic islands might become the seeds that allow sessile carbonate producers such as hermatypic colonial corals to colonize unconsolidated substrates.

Insights into the bacterial community compositions of peanut worm (Sipunculus nudus) and their association with the surrounding environment

The peanut worm (Sipunculus nudus) is an important economic and ecological fishery resource in China. To determine how S. nudus interacts with its culture environment, in this study, the diversity, composition, and interrelationships of bacterial communities in the intestine, coelomic fluid, and culture environment of S. nudus through high-throughput sequencing using the Illumina HiSeq sequencing platform. Alpha diversity analysis showed that the highest bacterial community richness and diversity were found in the sediments. Species annotation revealed that Proteobacteria dominated in all samples (48.92%–58.36%), and the other dominant phyla were highly variable, indicating a certain independence between the environment and the composition of the bacterial community of S. nudus. The coelomic fluid of S. nudus contained high concentrations of Bacillus and could be a source of potential probiotic bacteria for isolation and culture. Cluster and operational taxonomic unit Venn diagram analyses showed that the bacterial community composition in the intestine was more similar to that in the sediments compared with water. The predicted functional analysis of bacterial communities indicated that the functions of bacterial communities in the different surrounding environment were highly similar and involved in various aspects, such as membrane transport, amino acid metabolism, carbohydrate metabolism, and replication and repair. Our findings have important implications on the establishment of sustainable microecological regulation and management strategies for S. nudus culture and provide a reference for the development and utilization of probiotic bacteria.

Changes in the feeding habits of the bat ray Myliobatis californica (Gill 1865) during climatic anomalies off the west coast of the Baja California Peninsula, Mexico

The Mexican Pacific was influenced by “La Mancha” and “El Niño”, from 2014 and until 2016. The increase in sea surface temperature influenced the feeding habits of the bat ray (Myliobatis californica) in the northwest of Baja California Sur, Mexico. To evaluate possible changes in the diet, stomach content analysis and analysis of stable isotopes of carbon (δ13C) and nitrogen (δ15N) in muscle was performed during normal (2012 and 2013) and anomalous years (2014, 2015, and 2016). During the normal years, the main prey was the crab Dynomene spp. (% Prey Specific Relative Importance Index (PSIRI) = 29.3) and the stomatopod Hemisquilla californiensis (% PSIRI = 10.6). In contrast, during the anomalous years, these preys were replaced by the pelagic red crab Pleuroncodes planipes (% PSIRI = 28.5) and peanut worms Sipunculus spp. (% PSIRI = 7.9). During normal years the median isotopic values recorded were: δ13C = −16.2‰ and δ15N = 15.2‰. During the anomalous years, δ13C was −16.3‰ and δ15N was 15.1‰. Between the different periods no trophic (p (probability) > 0.05) or isotopic overlaps (p > 0.3) were found. The change in the diet of M. californica during the anomalous years is an adaptive response to the increase in water temperature caused by “La Mancha” and “El Niño”. The massive presence of P. planipes on the northwest coast of Baja California Sur is associated with the increase in water temperature, which makes P. planipes a food source for M. californica.

High-resolution imaging sheds new light on a multi-tier symbiotic partnership between a "walking" solitary coral, a sipunculan, and a bivalve from East Africa.

Marine symbioses are integral to the persistence of ecosystem functioning in coral reefs. Solitary corals of the species Heteropsammia cochlea and Heterocyathus aequicostatus have been observed to live in symbiosis with the sipunculan worm Aspidosiphon muelleri muelleri, which inhabits a cavity within the coral, in Zanzibar (Tanzania). The symbiosis of these photosymbiotic corals enables the coral holobiont to move, in fine to coarse unconsolidated substrata, a process termed as “walking.” This allows the coral to escape sediment cover in turbid conditions which is crucial for these light‐dependent species. An additional commensalistic symbiosis of this coral‐worm holobiont is found between the Aspidosiphon worm and the cryptoendolithic bivalve Jousseaumiella sp., which resides within the cavity of the coral skeleton. To understand the morphological alterations caused by these symbioses, interspecific relationships, with respect to the carbonate structures between these three organisms, are documented using high‐resolution imaging techniques (scanning electron microscopy and µCT scanning). Documenting multi‐layered symbioses can shed light on how morphological plasticity interacts with environmental conditions to contribute to species persistence.