Ascidian Species Research Articles

Harnessing Ascidians as Model Organisms for Environmental Risk Assessment

Environmental Risk Assessment (ERA) often relies on a restricted set of species as bio-indicators, introducing uncertainty when modeling complex environmental variables. This may lead to oversimplified or erroneous risk assessments. Ascidians, marine filter-feeding sessile chordates, are valuable models for scientific research in various biological fields such as stem cell biology, embryogenesis, regeneration, innate immunity, and developmental biology. Their global distribution, sensitivity to pollutants, high abundance, mass sexual reproduction, and habitation in coastal areas impacted by anthropogenic pollution make them excellent indicators for monitoring marine pollution and global environmental changes, including biological invasions and species diversity diminution cases. Despite their potential as environmental bioindicators, ascidians remain underutilized in ERAs (≤0.13% of ERA studies), particularly in the field of chemical pollution impact assessment, primarily due to a lack of standardization. This underrepresentation poses a challenge for accurate modeling, especially in models relying on a broad range of species (e.g., Species Sensitivity Distributions). Given these constraints, expanding the use of ascidians in ERAs could improve the comprehension and precision of environmental changes and their assessments. This underscores the necessity for future research to establish standardized testing protocols and choose the most suitable ascidian species for inclusion in ERAs.

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.

Do DOM quality and origin affect the uptake and accumulation of a lipid-soluble contaminant in a filter feeding ascidian species (Ciona) that can target small particle size classes?

The widely reported increase of terrestrial dissolved organic matter (terrDOM) in northern latitude coastal areas (“coastal darkening”) can impact contaminant dynamics in affected systems. One potential impact is based on differences in chemical adsorption processes of the molecularly larger terrDOM compared to marine DOM (marDOM) that leads to increased emulsification of lipophilic contaminants with terrDOM. Filter feeders filter large amounts of water and DOM daily and thus are directly exposed to associated contaminants through both respiration and feeding activity. Thus, increased exposure to terrDOM could potentially lead to an increase in bioaccumulation of lipid soluble contaminants in filter feeders. To assess the effect of DOM on bioaccumulation in filter feeders, we exposed the mucous based filter feeding ascidian Ciona intestinalis (formerly known as Ciona intestinalis Type B), to the lipophilic veterinary drug teflubenzuron (log KOW: 5.39) in combination with four DOM treatments: TerrDOM, marDOM, a mix of the two called mixDOM, and seawater without DOM addition. The exposure lasted for 15 days, after which the individuals in all DOM treatments showed a trend towards higher bioaccumulation of Teflubenzuron than those in the seawater control. However, there was considerable overlap in posterior distributions. Against our expectations, marDOM resulted in the highest bioaccumulation factor (BAF), followed by mixDOM, with terrDOM resulting in the lowest BAF except for seawater (kinetic BAF L/kg median, 2.5 %–97.5 % percentile marDOM 94, 74–118; mixDOM 82, 63–104; terrDOM 79; 61-99; seawater 61, 44–79). All BAFs were below the level of concern according to the EU REACH regulation (BAF < 2000 L / kg) and, therefore, likely not environmentally problematic in the examined context. However, the results show that DOM can act as a dietary vector; thus, different combinations of contaminants, DOM, and filter feeding organisms should be tested further.

A Dicopper(II)‐Based Carbonic Anhydrase Model—Quantum‐Chemical Evaluation of the Mechanistic Pathway

AbstractThe cyanobacterium Prochloron didemni, an obligate symbiont of different species of colonial ascidians, occurring in the Pacific and Indian Oceans, produces a variety of cyclic peptides. These patellamide‐type macrocycles lead to relatively stable dicopper(II) complexes that are extremely efficient carbonic anhydrase mimics, the most active model systems known so far. Importantly, it recently was shown that copper(II) is coordinated to patellamide derivatives in Prochloron cells. An interesting question therefore is, whether the biological function of patellamide‐type macrocycles is related to the catalytic activity in CO2 hydration or its reverse. Here, we present a computational study to evaluate the energetics of the catalytic cycle in search of a possible answer to these questions and compare the computed energy barriers with the experimental kinetic data. It emerges that release of the bridging carbonate is a critical step and that the catalysis product inhibits catalysis at pH values above approx. 7. Therefore, carbonate transport rather than CO2 hydrolysis is proposed as the biological function of copper(II)‐patellamide complexes in the Prochloron‐Ascidian symbiosis.

A Dicopper(II)-Based Carbonic Anhydrase Model-Quantum-Chemical Evaluation of the Mechanistic Pathway.

The cyanobacterium Prochloron didemni, an obligate symbiont of different species of colonial ascidians, occurring in the Pacific and Indian Oceans, produces a variety of cyclic peptides. These patellamide-type macrocycles lead to relatively stable dicopper(II) complexes that are extremely efficient carbonic anhydrase mimics, the most active model systems known so far. Importantly, it recently was shown that copper(II) is coordinated to patellamide derivatives in Prochloron cells. An interesting question therefore is, whether the biological function of patellamide-type macrocycles is related to the catalytic activity in CO2 hydration or its reverse. Here, we present a computational study to evaluate the energetics of the catalytic cycle in search of a possible answer to these questions and compare the computed energy barriers with the experimental kinetic data. It emerges that release of the bridging carbonate is a critical step and that the catalysis product inhibits catalysis at pH values above approx. 7. Therefore, carbonate transport rather than CO2 hydrolysis is proposed as the biological function of copper(II)-patellamide complexes in the Prochloron-Ascidian symbiosis.

Cymric, a Maternal and Zygotic HTK-16-Like SHARK Family Tyrosine Kinase Gene, Is Disrupted in Molgula occulta, a Tailless Ascidian.

AbstractWe describe the cloning and expression of a nonreceptor tyrosine kinase, cymric (Uro-1), a HTK-16-like (HydraTyrosineKinase-16) gene, identified in a subtractive screen for maternal ascidian cDNAs in Molgula oculata, an ascidian species with a tadpole larva. The cymric gene encodes a 4-kb mRNA expressed in gonads, eggs, and embryos in the tailed M. oculata but is not detected in eggs or embryos of the closely related tailless species Molgula occulta. There is a large insertion in cymric in the M. occulta genome, as shown by transcriptome and genome analyses, resulting in it becoming a pseudogene. The cymric amino acid sequence encodes a nonreceptor tyrosine kinase with an N-terminal region containing two SH2 domains and five ankyrin repeats, similar to the HTK-16-like gene found in other ascidians. Thus, the ascidian cymric genes are members of the SHARK (Src-homology ankyrin-repeat containing tyrosine kinase) family of nonreceptor tyrosine kinases, which are found throughout invertebrates and missing from vertebrates. We show that cymric is lacking the tyrosine kinase domain in the tailless M. occulta, although the truncated mRNA is still expressed in transcriptome data. This maternal and zygotic HTK-16-like tyrosine kinase is another described pseudogene from M. occulta and appears not to be necessary for adult development.

Molecular control of cellulosic fin morphogenesis in ascidians

BackgroundThe tunicates form a group of filter-feeding marine animals closely related to vertebrates. They share with them a number of features such as a notochord and a dorsal neural tube in the tadpole larvae of ascidians, one of the three groups that make tunicates. However, a number of typical chordate characters have been lost in different branches of tunicates, a diverse and fast-evolving phylum. Consequently, the tunic, a sort of exoskeleton made of extracellular material including cellulose secreted by the epidermis, is the unifying character defining the tunicate phylum. In the larva of ascidians, the tunic differentiates in the tail into a median fin (with dorsal and ventral extended blades) and a caudal fin.ResultsHere we have performed experiments in the ascidian Phallusia mammillata to address the molecular control of tunic 3D morphogenesis. We have demonstrated that the tail epidermis medio-lateral patterning essential for peripheral nervous system specification also controls tunic elongation into fins. More specifically, when tail epidermis midline identity was abolished by BMP signaling inhibition, or CRISPR/Cas9 inactivation of the transcription factor coding genes Msx or Klf1/2/4/17, median fin did not form. We postulated that this genetic program should regulate effectors of tunic secretion. We thus analyzed the expression and regulation in different ascidian species of two genes acquired by horizontal gene transfer (HGT) from bacteria, CesA coding for a cellulose synthase and Gh6 coding for a cellulase. We have uncovered an unexpected dynamic history of these genes in tunicates and high levels of variability in gene expression and regulation among ascidians. Although, in Phallusia, Gh6 has a regionalized expression in the epidermis compatible with an involvement in fin elongation, our functional studies indicate a minor function during caudal fin formation only.ConclusionsOur study constitutes an important step in the study of the integration of HGT-acquired genes into developmental networks and a cellulose-based morphogenesis of extracellular material in animals.

Temporospatial hierarchy and allele-specific expression of zygotic genome activation revealed by distant interspecific urochordate hybrids

Zygotic genome activation (ZGA) is a universal process in early embryogenesis of metazoan, when the quiescent zygotic nucleus initiates global transcription. However, the mechanisms related to massive genome activation and allele-specific expression (ASE) remain not well understood. Here, we develop hybrids from two deeply diverged (120 Mya) ascidian species to symmetrically document the dynamics of ZGA. We identify two coordinated ZGA waves represent early developmental and housekeeping gene reactivation, respectively. Single-cell RNA sequencing reveals that the major expression wave exhibits spatial heterogeneity and significantly correlates with cell fate. Moreover, allele-specific expression occurs in a species- rather than parent-related manner, demonstrating the divergence of cis-regulatory elements between the two species. These findings provide insights into ZGA in chordates.

Salinity-mediated limitation of asexual reproduction in the colonial ascidian Polyandrocarpa zorritensis

Ascidians are among the most common invasive marine invertebrates worldwide. Many species of non-indigenous ascidians (NIAs) have successfully colonized the Mediterranean Sea, notably within anthropized coastal lagoons and harbors. Although invasive species are generally characterized by their broad ecological tolerance, different ascidian species exhibit varied responses to biotic and abiotic environmental stressors, including temperature and salinity. Acquiring a better understanding about of the impact of such parameters on ascidian life history is crucial for predicting the invasive potential of NIAs. In this study, we investigated the impact of various salinities on the reproduction of the colonial ascidian Polyandorcarpa zorritensis, a species indigenous to Peru and a thriving invader. P. zorritensis undergoes asexual reproduction via a peculiar form of budding named vasal budding and produces resistant spherules, which likely facilitated its dissemination over long distances. Despite its widespread distribution along the Pacific and Atlantic coasts, it is only found in a few Mediterranean coastal areas with a low salinity. We tested the impact of different salinity conditions on the sexual and asexual reproduction rates of P. zorritensis in a controlled laboratory setting. Our experiments showed that the rate of asexual reproduction in colonies bred at 29 or 36 ppt salinity levels, corresponding to the natural range inhabited by P. zorritensis, was higher than those grown in 40 ppt salinity, commonly found in Mediterranean marinas and harbors. The results suggest that, although P. zorritensis has been present in the Mediterranean for several decades, its potential for invasion could be constrained by an intolerance to high salinity.

Graveyards of Giant Pandas at the Bottom of the Sea? A Strange-Looking New Species of Colonial Ascidians in the Genus &lt;i&gt;Clavelina&lt;/i&gt; (Tunicata: Ascidiacea)

Graveyards of Giant Pandas at the Bottom of the Sea? A Strange-Looking New Species of Colonial Ascidians in the Genus &lt;i&gt;Clavelina&lt;/i&gt; (Tunicata: Ascidiacea)

Supplemental re-description of a deep-sea ascidian, Fimbrora calsubia (Ascidiacea, Enterogona), with an inference of its phylogenetic position

Fimbrora Monniot &amp; Monniot, 1991, a macrophagous ascidian genus within the family Ascidiidae Adams &amp; Adams, 1858, is currently monotypic, represented by F. calsubia Monniot &amp; Monniot, 1991, a species previously recorded from the bottom of the South Pacific at depths of 1000–1860 m. The taxonomic status of Fimbrora has remained ambiguous because characteristics in its branchial papillae and neural-gland opening are incompletely known in previous studies, while these traits are essential for distinguishing other ascidiid genera. So far, no nucleotide sequence representing F. calsubia is available. In this study, we collected a single specimen of F. calsubia at a depth of 2027 m, about 400 km off the Pacific coast of Honshu, Japan. This is the deepest record, as well as the first report from the North Pacific, for the species. Our examination indicates that Fimbrora is morphologically similar to another ascidiid genus, Psammascidia Monniot, 1962, by having only secondary branchial papillae in the pharynx. Our phylogenetic analysis, based on the 18S ribosomal RNA and cytochrome c oxidase subunit I genes, along with those of 27 ascidian species available in public databases, showed that F. calsubia was more closely related to Ascidia zara Oka, 1935, Phallusia fumigata (Grube, 1864) and Phallusia mammilata (Cuvier, 1815) than to Ascidia ceratodes (Huntsman, 1912), Ascidiella aspersa (Müller, 1776) and Ascidiella scabra (Müller, 1776). Our results also indicated that acquisitions of macrophagous feeding by deep-sea members happened independently at least three times in the evolutionary history of the entire Ascidiacea.

The distribution of the red-throated ascidian Herdmania momus shifts northwards in association with ocean warming in the Korean Peninsula

Abstract Climate change poses a major threat to marine ecosystems, with its effects felt worldwide. A major effect of climate change on marine ecosystems is the rise in water temperature, leading to a northward expansion of habitats for marine organisms. Herdmania momus, a species of ascidians (sea squirts), originally found in tropical and subtropical regions, was introduced to the Korean Peninsula. In this study, we examined the habitat of H. momus along the southeastern coast of the Korean Peninsula between 2016 and 2022. We found that H. momus settlements were observed across the entire survey area, with confirmed habitation in Busan in 2016, Ulsan in 2021, and Gyeongju (the northernmost location) in 2022. The observed habitation trend indicates a rapid geographical expansion, occurring approximately 79 years earlier than previously predicted. These observations demonstrate that marine organisms are undergoing a more rapid geographical expansion than previously projected. These unexpected findings should inform government policies related to proactive measures and strategies for managing the impact of climate change on marine ecosystems.

Major ascidian species with negative impacts on bivalve aquaculture: Current knowledge and future research aims

Major ascidian species with negative impacts on bivalve aquaculture: Current knowledge and future research aims

Seven ascidian (Tunicata) species from the North Eastern Mediterranean

Members of the tunicates, a subphylum of marine filter-feeder chordates, inhabit all marine and oceanic habitats from the subtidal to the abyssal. Considered the closest relatives to the vertebrates, the tunicates are widely used as model organisms for evo-devo, allorecognition, senescence, and whole-body regeneration studies. However, species boundaries are poorly understood due to the high morphological and genetic plasticity that characterizes many tunicate taxa. Here, we present findings on seven tunicate species (Botrylloides israeliense, Botrylloides sp., Botryllus humilis, Botryllus schlosseri, Symplegma brakenhielmi, Polyclinum constellatum and Didemnum perlucidum) sampled from six Turkish sites at the North Eastern Mediterranean Sea and employed the mitochondrial barcoding marker (COI) for evaluating the relationships among geographically restricted and widely spread ascidian species. Species delimitation was conducted using sequences generated in the current study in addition to sequences obtained from GenBank. General morphological features and colors of colonies were recorded at sampling sites. Then, all Styelidae colonies were attached and cultured on slides in an aquaculture room, enabling the study of other features, such as zooid distributions and sizes, oral tentacle numbers, and life cycles, using stereo and light microscopes. The spicules of formalin-fixed Didemnum perlucidum samples were examined under a light microscope. Then, scientific names were assigned to all species based on the results of the species delimitation and on comparisons of the obtained COI sequences with GenBank sequences. A putative new Botrylloides species (Botrylloides sp.) from the Antalya region was revealed, with a 99% match with the COI gene of another specimen from Saudi Arabia; further waiting for detailed traditional taxonomy.

Ascidian diversity and abundance in North Carolina seagrass meadows

AbstractAscidians are sessile marine invertebrates found all over the world in a variety of natural and artificial habitats. The objective of this study was to provide the first inventory of ascidian diversity and abundance in North Carolina (NC) seagrass meadows. Eight sites along the NC coast were surveyed in May and June 2021 and at each site, 20 quadrats were deployed. All ascidian species within the quadrats were counted and identified based on morphological characterization and sequencing of the barcoding gene. Seagrass percent cover, biomass, and shoot density were also quantified. Ascidians were found in six sites and four species were recorded: the solitary species Molgula manhattensis, Styela plicata, and Bostrichobranchus sp., and the colonial Didemnum lutarium. Colonial specimens were generally attached to hard substrate, while solitary species were often found attached to seagrass rhizomes and leaves. S. plicata and M. manhattensis haplotype diversity were equivalent to previous descriptions from individuals collected on artificial substrates and other locations. Ascidian abundance was positively correlated with seagrass biomass but not with seagrass percent cover. A fifth species, the solitary Styela canopus, appeared in a site revisited in October 2021. Temperate ascidians are known to have seasonal cycles, thus further research should include monthly surveys to assess ascidian diversity and abundance over time.

Seasonal distribution of solitary and colonial benthic ascidians (Chordata) with ecological gradients in and around Andaman and Nicobar Islands, India

AbstractAscidians are rapidly emerging biofouling organisms that generally occur on the natural and artificial substratum of the marine environment. However, very few studies have been reported on their species diversity and distribution patterns. This present investigation focuses on the seasonal impact on ascidians diversity. The distributions of ascidians were recorded by the collection of samples in six sampling stations from Andaman and Nicobar Islands. A total of 43 species of ascidians (32 colonial, 11 solitary) were recorded and most of the species are under the Didemnidae family. Various diversity indexes were utilized to examine the diversity, richness and evenness of species and to look at the assorted variety among tests and their likeness regarding species composition. The greatest (3.227) individuals were observed during summer season in Pigeon Island and the lowest (2.063) number of species were recorded during the monsoon season in Haddowharf harbour. The Margalef richness (d) fluctuated between in the range of 2.033 and 5.53 during the monsoon in Haddowharf harbour and Carbyns Cove beach in summer seasons. The Pielou's evenness record (J′) changed from 0.7409 to 0.9713. Diversity indexes were recorded as followed the order of 0.9642, 0.9552, 0.9482 and 0.9292 at Carbyns Cove shoreline, Burmanella, Pigeon Island and Campbell Bay individually during summer season. Present findings in the cluster examination uncovered that the species at various stations have framed four gatherings. Most elevated species rates were spread in summer. Field observation shows that species distribution is able to tolerate temperature ranges of 26.9–30.3°C and salinity of 32.1–34.64 ppt. Occasional examples of space occupation by colonial ascidians corresponded with seasonal changes in seawater temperature and salinity.

Topological properties and shape of proliferative and nonproliferative cell monolayers.

During embryonic development, structures with complex geometry can emerge from planar epithelial monolayers; studying these shape transitions is of key importance for revealing the biophysical laws involved in the morphogenesis of biological systems. Here, using the example of normal proliferative monkey kidney (COS) cell monolayers, we investigate global and local topological characteristics of this model system in dependence on its shape. The obtained distributions of cells by their valence demonstrate a difference between the spherical and planar monolayers. In addition, in both spherical and planar monolayers, the probability of observing a pair of neighboring cells with certain valences depends on the topological charge of the pair. The zero topological charge of the cell pair can increase the probability for the cells to be the nearest neighbors. We then test and confirm that analogous relationships take place in a more ordered spherical system with a larger fraction of 6-valent cells, namely, in the nonproliferative epithelium (follicular system) of ascidian species oocytes. However, unlike spherical COS cell monolayers, ascidian monolayers are prone to nonrandom agglomeration of 6-valent cells and have linear topological defects called scars and pleats. The reasons for this difference in morphology are discussed. The morphological peculiarities found are compared with predictions of the widely used vertex model of epithelium.

Comparative Transcriptomic Analysis Reveals the Functionally Segmented Intestine in Tunicate Ascidian.

The vertebrate intestinal system consists of separate segments that remarkably differ in morphology and function. However, the origin of intestinal segmentation remains unclear. In this study, we investigated the segmentation of the intestine in a tunicate ascidian species, Ciona savignyi, by performing RNA sequencing. The gene expression profiles showed that the whole intestine was separated into three segments. Digestion, ion transport and signal transduction, and immune-related pathway genes were enriched in the proximal, middle, and distal parts of the intestine, respectively, implying that digestion, absorption, and immune function appear to be regional specializations in the ascidian intestine. We further performed a multi-species comparison analysis and found that the Ciona intestine showed a similar gene expression pattern to vertebrates, indicating tunicates and vertebrates might share the conserved intestinal functions. Intriguingly, vertebrate pancreatic homologous genes were expressed in the digestive segment of the Ciona intestine, suggesting that the proximal intestine might play the part of pancreatic functions in C. savignyi. Our results demonstrate that the tunicate intestine can be functionally separated into three distinct segments, which are comparable to the corresponding regions of the vertebrate intestinal system, offering insights into the functional evolution of the digestive system in chordates.

The impact of ascidian biofouling on the farmed Mediterranean mussel Mytilus galloprovincialis physiology and welfare, revealed by stress biomarkers

In biofouling communities, ascidians are among the most damaging species, presenting severe threats, such as depressed growth rates and decreased chances of lower survival, to shellfish aquaculture. However, little is known concerning the fouled shellfish physiology. In an effort to obtain information for the magnitude of stress caused by ascidians to farmed Mytilus galloprovincialis, five seasonal samplings took place in a mussel aquaculture farm suffering from ascidian biofoulants, in Vistonicos Bay, Greece. The dominant ascidian species were recorded and several stress biomarkers, including Hsp gene expression at both mRNA and protein levels, as well as MAPKs levels, and enzymatic activities of intermediate metabolism were examined. Almost all investigated biomarkers revealed elevated stress levels in fouled mussels compared to non-fouled. This enhanced physiological stress seems to be season-independent and can be attributed to the oxidative stress and/or feed deprivation caused by ascidian biofouling, thus illuminating the biological impact of this phenomenon.

Phylogeographic and Morphological Analysis of Botrylloides niger Herdman, 1886 from the Northeastern Mediterranean Sea

Botrylloides niger (class Ascidiacea) is an invasive marine filter-feeding invertebrate that is believed to originate from the West Atlantic region. This species of colonial tunicate has been observed in several locations along the coasts of Israel and around the Suez Canal, but it has not yet been reported on the coasts of the Northeastern Mediterranean Sea (NEMS), suggesting an ongoing Lessepsian migration. However, the extent of this invasion might be concealed by reports of other potentially misidentified species of Botrylloides, given that the strong morphological similarities within this genus renders taxonomical identification particularly challenging. In this study, we performed a phylogeographic and morphological analysis of B. niger in the NEMS. We collected 238 samples from 8 sampling stations covering 824 km of the coastlines of NEMS. We reported 14 different morphotypes, of which the orange-brown, orange, and brown-striped morphs were the most abundant. Using the mitochondrial cytochrome C oxidase I (COI) as a DNA barcode marker, we identified 4 haplotypes. The COI haplotypes clustered with the reference B. niger sequences from GenBank and differed significantly from the sister Botrylloides species. We confirmed our identification using three additional barcoding markers (Histone 3, 18S rRNA, and 28S rRNA), which all matched with over 99% similarity to reference sequences. In addition, we monitored a station for a year and conducted a temporal analysis of the collected colonies. The colonies were absent during the winter and spring, while new colonies were established in the summer and expanded during autumn. We performed demographic population analysis on our spatial data that identified a possible population subdivision at a sampling site, which might have been caused by local freshwater input. Herein, we present the first report on the presence of Botrylloides niger in the NEMS. This study represents a key step toward understanding the diversity and the propagation of this highly invasive species of colonial ascidians, both within the Mediterranean basin as well as globally.