Ascidian Styela Plicata Research Articles

Effects of biological filtration by ascidians on microplastic composition in the water column

Plastic pollution, a widespread environmental challenge, significantly impacts marine ecosystems. The degradation of plastic under environmental conditions results in the generation of microplastic (MP; <5 mm) fragments, frequently ingested by marine life, including filter-feeders such as ascidians (Chordata, Ascidiacea). These organisms are integral to benthic-pelagic coupling, transporting MP from the water column through marine food web.Here, we explored the effect of filtration and digestion by the solitary ascidian Styela plicata on the composition of MP in the water column and on the sinking rates of faecal matter, focusing on differences between two distinct plastics, polystyrene (PS) and the biodegradable polylactic acid (PLA). The ascidians efficiently removed 2–5 μm particles within 2 h of filtration. Following digestion and secretion process, PS concentrations in water increased while PLA concentration remained stable. Some particles were egested into the water column repackaged inside faecal pellets, which significantly increased the pellets' drag force and sinking velocity. Raman spectral analysis of digested MP revealed distinct spectrum alterations due to coating by organic substances. These findings highlight the role of ascidians — and other filter-feeders— in modifying the structure of MP in their environment. Research into such modifications is crucial for understanding the MP cycle and its consequences in marine environments.

The past, the recent, and the ongoing evolutionary processes of the worldwide invasive ascidian Styela plicata.

Invasive species are one of the main threats to global biodiversity and, within marine ecosystems, tunicates feature some prominent examples. Styela plicata is an ascidian species inhabiting harbours in all temperate oceans and seas, thus being considered a thriving invasive species. However, this species' adaptive mechanisms, introduction history, and population structure have never been completely elucidated. Here, by genotyping 87 S. plicata individuals from 18 localities worldwide with 2b-RADseq, we confirm the global presence of four chromosome inversions, demonstrate population structuring on this species, detect local adaptation signals, and infer historical demographic events. We show that North Carolina individuals constitute an unrelated population, Atlanto-Mediterranean and Pacific localities form their own genetic clusters with substructuring, being the most evident the split between northern and southern Atlantic localities. The locality of South Carolina presents an intermediate genetic position between North Carolina and the other two groups pointing to a hybrid origin with recurrent gene flow. We generate and test demographic models, providing evidence of two independent introduction events to the Atlantic and Pacific, and an admixture that originated the population of South Carolina. Finally, we identify candidate loci for adaptation, with functions involved with cell processes, metabolism, development, and ion transport, among others. Overall, this study highlights the complex historical processes of S. plicata, which have led this species to its current distribution, population structure, and local adaptation footprint in oceans worldwide.

Identification of astrocyte-like cells in an adult ascidian during regeneration of the central nervous system.

The mechanisms underlying regeneration of the central nervous system (CNS) following lesions have been studied extensively in both vertebrate and invertebrate models. To shed light on regeneration, ascidians, a sister group of vertebrates and with remarkable ability to regenerate their brains, constitute an appropriate model system. Glial cells have been implicated in regeneration in vertebrates; however, their role in the adult ascidian CNS regeneration is unknown. A model of degeneration and regeneration using the neurotoxin 3-acetylpyridine (3AP) in the brain of the ascidian Styela plicata was used to identify astrocyte-like cells and investigate their role. We studied the CNS of control ascidians (injected with artificial sea water) and of ascidians whose CNS was regenerating (1 and 10 days after the injection with 3AP). Our results show that the mRNA of the ortholog of glutamine synthetase (GS), a glial-cell marker in vertebrates, is increased during the early stages of regeneration. Confirming the identity of GS, the protein was identified via immunostaining in a cell population during the same regeneration stage. Last, a single ortholog of GS (GSII) is present in ascidian and amphioxus genomes, while two types exist in fungi, some invertebrates, and vertebrates, suggesting that ascidians have lost the GSI type. Taken together, our findings revealed that a cell population expressing glial-cell markers may play a role in regeneration in adult ascidians. This is the first report of astrocyte-like cells in the adult ascidian CNS, and contributes to understanding of the evolution of glial cells among metazoans.

Lake faro (Central Mediterranean): A potential short-term reservoir for Pinna nobilis

The noble fan shell, Pinna nobilis, is an endemic Mediterranean species included in the IUCN Red List because it is at risk of extinction after the global mass mortality event (MME) first detected in late 2016 in the Spanish waters. Throughout the whole Mediterranean Sea, measures to evaluate reproduction and recruitment in relict populations have been undertaken, in view of a wished species recovery. In this study we highlight the potential resilience of the Lake Faro P. nobilis population (north-eastern Sicily), which partially survived to the MME affection. Results of both monitoring and installation of larval collectors during a three-year period (from 2021 to 2023) are reported. Despite spawning events show that the population is reproductively active, a poor natural recruitment was detected. Each sign of potential resilience, moreover, contrasts with the failure of collectors in capturing P. nobilis larvae, while they were highly colonized by more competitive organisms, including non-native species, such as the ascidian Styela plicata and the bivalves Pinctada radiata, Isognomon bicolor and Anadara transversa.

Multidimensional variability of the microbiome of an invasive ascidian species

Animals, including invasive species, are complex entities consisting of a host and its associated symbionts (holobiont). The interaction between the holobiont components is crucial for the host's survival. However, our understanding of how microbiomes of invasive species change across different tissues, localities, and ontogenetic stages, is limited. In the introduced ascidian Styela plicata, we found that its microbiome is highly distinct and specialized among compartments (tunic, gill, and gut). Smaller but significant differences were also found across harbors, suggesting local adaptation, and between juveniles and adults. Furthermore, we found a correlation between the microbiome and environmental trace element concentrations, especially in adults. Functional analyses showed that adult microbiomes possess specific metabolic pathways that may enhance fitness during the introduction process. These findings highlight the importance of integrated approaches in studying the interplay between animals and microbiomes, as a first step toward understanding how it can affect the species' invasive success.

Can stand the heat – ecology of the potentially invasive ascidian Styela plicata along the Mediterranean coast of Israel

IntroductionBoth colonial and solitary ascidians (Chordata, Ascidiacea) have been gathering attention due to their impact on coastal environments. The solitary ascidian Styela plicata was first observed along the coast of Israel in 2014. It is considered a cosmopolitan species of unknown origin, and is well-known for its ability to create large aggregations on artificial substrates in the western Mediterranean Sea, North-Pacific Ocean and the Atlantic. The extreme environmental conditions that characterize the Mediterranean coast of Israel, with a salinity of 40 psu and sea-surface temperature reaching 33°C, may however present a barrier to its future spread.MethodsWe examined the seasonal reproduction patterns and population structure of three populations over two years, in addition to spatial surveys and monitoring oocyte release under laboratory conditions.ResultsHistological sections revealed significant differences in reproduction patterns between seasons and sites, with a higher proportion of the larger oocytes size group observed during the winter for all sites. Along the Israeli coast, with the exception of one site, S. plicata is currently restricted to marinas and ports.DiscussionWith an impressive ability to survive the extreme environmental conditions and sustain reproductive populations. Acquiring data on its spread and establishment is essential for modeling S. plicata’s distribution under the current and predicted global climate changes, and for developing tools for its mitigation and control.

Morpho-structural adaptations of the integument in different aquatic organisms

The integument acts as a barrier to protect the body from harmful pathogenic infectious agents, parasites, UV rays, trauma, and germs. The integument of invertebrates and vertebrates are structurally different: while invertebrates usually have a simple monolayer epidermis frequently covered by mucus, cuticles, or mineralized structures, vertebrates possess a multilayered epidermis with several specialized cells. This study aims to describe by morphological, histological, and immunohistochemical analyses, the morpho-structural adaptations throughout evolution of the integument of gastropod Aplysia depilans (Gmelin, 1791), ascidian Styela plicata (Lesuer, 1823), myxine hagfish Eptatretus cirrhatus (Forster, 1801) and teleost Heteropneustes fossilis (Bloch, 1794) for the first time, with special reference to sensory epidermal cells. Different types of cells could be identified that varied according to the species; including mucous cells, serous glandular cells, clavate cells, club cells, thread cells, and support cells. In all integuments of the specimens analyzed, sensory solitary cells were identified in the epidermis, immunoreactive to serotonin and calbindin. Our study provided an essential comparison of integuments, adding new information about sensory epidermal cells phylogenetic conservation and on the structural changes that invertebrates and vertebrates have undergone during evolution.

Fouling Community on Pinna nobilis Larval Collectors in the Adriatic—Impact of Invasive Species

In the last few years, the endemic Mediterranean bivalve Pinna nobilis has been exposed to dramatic mortality in its entire area, which could lead to the extinction of the species. Throughout the Mediterranean, a lot of effort is being put into finding ways of preserving it. One of the methods used to monitor recruitment and juveniles’ survival is the installation of collectors for bivalve larvae. We installed collectors at two locations: in Brijuni National Park (North Adriatic) and Luka Cove (central Adriatic). Our aim was to compare the fouling community on the collectors in two consecutive years (2019 and 2020), especially because the installation of collectors in 2020 coincided with mass mortality events of P. nobilis in the area. The number and size of juvenile P. nobilis and the qualitative and quantitative composition of the fouling communities were determined. The results show a reduction in the number and size of juvenile P. nobilis and an explosion of the invasive bivalve Anadara transversa population on collectors in the second year. In Luka Cove, another invasive species—the ascidian Styela plicata—also seriously affected other organisms on the collectors to the point of preventing analysis of the fouling community.

Facilitation of non-indigenous ascidian by marine eco-engineering interventions at an urban site

Marine artificial structures often support lower native species diversity and more non-indigenous species (NIS), but adding complex habitat and using bioreceptive materials have the potential to mitigate these impacts. Here, the interacting effects of structural complexity (flat, complex with pits) and concrete mixture (standard, or with oyster shell or vermiculite aggregate) on recruitment were assessed at two intertidal levels at an urban site. Complex tiles had less green algal cover, oyster shell mixtures had less brown (Ralfsia sp.) algal cover. At a low tidal elevation, the non-indigenous ascidian Styela plicata dominated complex tiles. Additionally, mixtures with oyster shell supported higher total cover of sessile species, and a higher cover of S. plicata. There were no effects of complexity or mixture on biofilm communities and native and NIS richness. Overall, these results suggest that habitat complexity and some bioreceptive materials may facilitate colonisation by a dominant invertebrate invader on artificial structures.

Effects of Dermatan Sulfate from Marine Invertebrate Styela plicata in the Wound Healing Pathway: A Natural Resource Applied to Regenerative Therapy.

Acute and chronic dermatological injuries need rapid tissue repair due to the susceptibility to infections. To effectively promote cutaneous wound recovery, it is essential to develop safe, low-cost, and affordable regenerative tools. Therefore, we aimed to identify the biological mechanisms involved in the wound healing properties of the glycosaminoglycan dermatan sulfate (DS), obtained from ascidian Styela plicata, a marine invertebrate, which in preliminary work from our group showed no toxicity and promoted a remarkable fibroblast proliferation and migration. In this study, 2,4-DS (50 µg/mL)-treated and control groups had the relative gene expression of 84 genes participating in the healing pathway evaluated. The results showed that 57% of the genes were overexpressed during treatment, 16% were underexpressed, and 9.52% were not detected. In silico analysis of metabolic interactions exhibited overexpression of genes related to: extracellular matrix organization, hemostasis, secretion of inflammatory mediators, and regulation of insulin-like growth factor transport and uptake. Furthermore, in C57BL/6 mice subjected to experimental wounds treated with 0.25% 2,4-DS, the histological parameters demonstrated a great capacity for vascular recovery. Additionally, this study confirmed that DS is a potent inducer of wound-healing cellular pathways and a promoter of neovascularization, being a natural ally in the tissue regeneration strategy.

Effect of Predation on Fouling Communities in an Italian Hotspot of Non-Indigenous Species

The role of native predators in controlling or facilitating non-indigenous species (NIS) growing in a fouling assemblage was investigated with a 70-day caging experiment in a marina inside the Gulf of La Spezia (Italy). Submerged PVC panels were divided into three experimental treatment levels: “control panels”, “open-caged panels” and “caged panels”, to assess the predation effect on the fouling assemblages growing on the panels. Predation effect was observed for two investigated variables. The Shannon Index was significantly higher in caged panels in comparison with controls, while an opposite trend was observed for the coverage area of native species. When testing single NIS abundance, both facilitation and limitation of individual NIS settlement has been observed. Mean coverage of the sabellid Branchiomma luctuosum was significantly higher in caged than in uncaged or open-caged panels, while the ascidian Styela plicata was prevalent in open-caged panels. In general, a predation effect was observed when considering the whole fouling assemblage, including different trends in NIS coverage among treatment levels. The results of this first attempt performed in the Mediterranean Sea encourage further experiments to investigate the biotic resistance provided by native predators against NIS.

Central nervous system regeneration in ascidians: cell migration and differentiation.

Adult ascidians have the capacity to regenerate the central nervous system (CNS) and are therefore excellent models for studies on neuroregeneration. The possibility that undifferentiated blood cells are involved in adult neuroregeneration merits investigation. We analyzed the migration, circulation, and role of hemocytes of the ascidian Styela plicata in neuroregeneration. Hemocytes were removed and incubated with superparamagnetic iron oxide nanoparticles (SPION), and these SPION-labeled hemocytes were injected back into the animals (autologous transplant), followed by neurodegeneration with the neurotoxin 3-acetylpyridine (3AP). Magnetic resonance imaging showed that 1, 5, and 10days after injury, hemocytes migrated to the intestinal region, siphons, and CNS. Immunohistochemistry revealed that the hemocytes that migrated to the CNS were putative stem cells (P-element-induced wimpy testis + or PIWI + cells). In the cortex of the neural ganglion, migrated hemocytes started to lose their PIWI labeling 5days after injury, and 10days later started to show β-III tubulin labeling. In the neural gland, however, the hemocytes remained undifferentiated during the entire experimental period. Transmission electron microscopy revealed regions in the neural gland with characteristics of neurogenic niches, not previously reported in ascidians. These results showed that migration of hemocytes to the hematopoietic tissue and to the 3AP-neurodegenerated region is central to the complex mechanism of neuroregeneration.

Invasive species fouling Perna perna (Bivalvia: Mytilidae) mussel farms

Invasive, fouling species increase management costs and reduce mussel growth, which jeopardizes mariculture. We studied the distribution of eight invasive species in Santa Catarina, the leading mussel producer in Brazil. Our goals were to determine their spatial distribution and prevalence on farm structures (buoys, long lines, and mussel socks), as well as understand the relevance of propagule pressure (recruitment), port distance, and area of the farm in this distribution. Although present in all sites, adult and recruits distribution were spatially restricted, showing that species might have a metapopulation structure. The most prevalent species were the ascidian Styela plicata, the barnacle Megabalanus coccopoma, the bryozoan Schizoporella errata, and the polychaete Branchiomma luctuosum. Recruitment was the main driver of three species distribution while distance to port explained only one species distribution. Based on those results, we discuss policy options, management, and regulation enforcement, that can be used in the mussel aquaculture elsewhere.

Bi-Hemispheric Distribution and Ecology of the Commensal Amphipod Leucothoe nagatai Ishimaru, 1985 (Crustacea: Leucothoidae)1

Reports on the taxonomy and distribution of the amphipod crustacean Leucothoe nagatai Ishimaru, 1985, are discussed including significant range extensions from the type locality in the Sea of Japan. This research was based on existing museum specimens, and in situ collections of new material. Originally described from the branchial chamber of the solitary ascidian Styela plicata (Lesueur, 1823), several new ascidian and sponges host species from California and New Zealand are documented for L. nagatai. Molecular studies confirm that northern and southern hemisphere populations are genetically indistinguishable. Color patterns in live and freshly preserved specimens of L. nagatai are diagnostic across its geographic range aiding in identification by nontaxonomists. Feeding habits of leucothoids within their invertebrate hosts are elucidated and discussed. Figures and plates of known invertebrate hosts, color patterns, and taxonomic structures are included.

Anthropogenic Sound in the Sea: Are Ascidians Affected?

Sound pollution in the marine environment has been increasing largely due to anthropogenic sources such as vessel traffic, coastal development, fossil fuel extraction, and military exercises. Studies determining the effects of anthropogenic sound on marine organisms have mostly focused on vertebrates, namely fish and marine mammals; however, less research has been done to study the effects of sound on marine invertebrates. The goal of this research is to examine the effect of anthropogenic sound on the ascidian Styela plicata (Lesueur, 1823) in Tampa Bay, FL. A total of 48 ascidians were collected from 2 sites with differing amounts of boat traffic and thus different anthropogenic soundscapes. Ascidians were individually exposed to 3 separate stimuli: a recording of a boat motor, a song recording, and a water current to simulate turbulence. Ascidian reactions were recorded as the frequency of siphon closing events and the length of time the siphons remained closed after disturbance. Ascidians from both sites increased the frequency and longevity of siphon closure events in response to anthropogenic stimuli but showed only a minor difference in response between sample sites. Research into the effect of anthropogenic sound on invertebrates such as ascidians and their symbionts may provide a better understanding of larger scale ecological impacts from such disturbances.

Mast cells and complement system: Ancient interactions between components of innate immunity.

The emergence and evolution of the complement system and mast cells (MCs) can be traced back to sea urchins and the ascidian Styela plicata, respectively. Acting as a cascade of enzymatic reactions, complement is activated through the classical (CP), the alternative (AP), and the lectin pathway (LP) based on the recognized molecules. The system's main biological functions include lysis, opsonization, and recruitment of phagocytes. MCs, beyond their classic role as master cells of allergic reactions, play a role in other settings, as well. Thus, MCs are considered as extrahepatic producers of complement proteins. They express various complement receptors, including those for C3a and C5a. C3a and C5a not only activate the C3aR and C5aR expressing MCs but also act as chemoattractants for MCs derived from different anatomic sites, such as from the bone marrow, human umbilical cord blood, or skin in vitro. Cross talk between MCs and complement is facilitated by the production of complement proteins by MCs and their activation by the MC tryptase. The coordinated activity between MCs and the complement system plays a key role, for example, in a number of allergic, cutaneous, and vascular diseases. At a molecular level, MCs and complement system interactions are based on the production of several complement zymogens by MCs and their activation by MC-released proteases. Additionally, at a cellular level, MCs act as potent effector cells of complement activation by expressing receptors for C3a and C5a through which their chemoattraction and activation are mediated by anaphylatoxins in a paracrine and autocrine fashion.

Putative stem cells in the hemolymph and in the intestinal submucosa of the solitary ascidian Styela plicata

BackgroundIn various ascidian species, circulating stem cells have been documented to be involved in asexual reproduction and whole-body regeneration. Studies of these cell population(s) are mainly restricted to colonial species. Here, we investigate the occurrence of circulating stem cells in the solitary Styela plicata, a member of the Styelidae, a family with at least two independent origins of coloniality.ResultsUsing flow cytometry, we characterized a population of circulating putative stem cells (CPSCs) in S. plicata and determined two gates likely enriched with CPSCs based on morphology and aldehyde dehydrogenase (ALDH) activity. We found an ALDH + cell population with low granularity, suggesting a stem-like state. In an attempt to uncover putative CPSCs niches in S. plicata, we performed a histological survey for hemoblast-like cells, followed by immunohistochemistry with stem cell and proliferation markers. The intestinal submucosa (IS) showed high cellular proliferation levels and high frequency of undifferentiated cells and histological and ultrastructural analyses revealed the presence of hemoblast aggregations in the IS suggesting a possible niche. Finally, we document the first ontogenetic appearance of distinct metamorphic circulatory mesenchyme cells, which precedes the emergence of juvenile hemocytes.ConclusionsWe find CPSCs in the hemolymph of the solitary ascidian Styela plicata, presumably involved in the regenerative capacity of this species. The presence of proliferating and undifferentiated mesenchymal cells suggests IS as a possible niche.

Exotic species dominate marinas between the two most populated regions in the southwestern Atlantic Ocean

Human occupation of coastal areas promotes the establishment of non-native species but information on bioinvasions is usually biased toward the Northern Hemisphere. We assessed non-native species' importance in sessile communities at six marinas along the most urbanized area of the Southwestern Atlantic coastline. We found 67 species, of which 19 are exotic. The most frequent species was the exotic polychaete Branchiomma luctuosum, while the most abundant was the exotic bryozoan Schizoporella errata that monopolized the substrata in three marinas. Along with S. errata, the exotic polychaete Hydroides elegans and ascidian Styela plicata dominated space in the three remaining marinas, while native species were in general rare. We show that communities associated with artificial substrata along this Brazilian urbanized area are dominated by exotic species and that using abundance data along with species identity can improve our understanding of the importance of exotic species for the dynamics of biological communities.

Dynamics of a biofouling community in finfish aquaculture: a case study from the South Adriatic Sea

Biofouling is one of the challenges that can strongly affect the finfish farm economy. Although several studies on biofouling in aquaculture have been conducted in the Mediterranean Sea, they focused on specific taxa or were limited to a particular period of sampling. The present study investigated for the first time the development, composition and variation in a biofouling community in a finfish farm with immersion time, season and depth. The results indicate that all these factors influence biofouling succession and recruitment. Moreover, the species that had a crucial role in structuring the community and in the farm cleaning activities were the ascidian Styela plicata and the bivalve Mytilus galloprovincialis. Compared with the literature data, the results highlight the heterogeneity in the composition of the biofouling present in the Mediterranean Sea. Moreover, such knowledge of the biofouling community could provide important information about management efforts and the costs that farmers will face when siting new fish farms.

Acute exposure to water-soluble fractions of marine diesel oil: Evaluation of apoptosis and oxidative stress in an ascidian

To understand the mechanisms involved in organisms’ responses to toxicity from oil pollution, we studied the effect of acute exposure (24 h) to the marine water-soluble fraction of diesel oil (WFDO) on the ascidian Styela plicata. We evaluated the mortality and behavior by means of the siphon reflex, and the response of blood cells (hemocytes) contained in the pharynx, by means of the production of nitric oxide (NO) and reactive oxygen species (ROS), in addition to the activity of the antioxidant enzyme catalase (CAT). We also correlated oxidative stress with the activation of apoptotic pathways. No mortality occurred 24 h after the ascidians were exposed to 5% and 10% marine WFDO; however, the siphon reflex, a behavioral test based on the time that the animals took to close their siphons, increased. We also observed an inflammatory response, as estimated by the increase in the number of hemocytes in the pharynx. NO and ROS production and CAT activity were reduced, whereas caspase-3, a signaling molecule involved in apoptosis, was activated. This suggests that in ascidians acutely exposed to oil, another mechanism can occur in addition to oxidative stress. Another possibility is that WFDO may directly interact with cellular macromolecules and activate caspase-3, independently of generating oxidative stress. The results showed that components of diesel oil affected a marine organism, which showed reduced ROS production in the pharynx cells, including hemocytes, and activation of apoptotic pathways.