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The aim of this study was to investigate the presence of alien phytoplankton species transported through ballast water of ships that docked on the Amazon coast. Phytoplankton samples were collected from 25 ships between 2012 and 2014, revealing 215 identified species, mostly comprising oceanic planktonic marine species. However, several coastal and freshwater species not yet documented on the Maranhão coast were also observed. The identification of several coastal and freshwater species not yet recorded for Amazonian environments in the ballast water of the Ponta da Madeira Maritime Terminal (TMPM), as well as toxic microalgae genera such as the dinoflagellates Alexandrium and Gymnodinium and of some diatom species from the genus Pseudo-nitzchia, raises concerns regarding the possibility of introducing species. This indicates that ballast water can be responsible for the introduction of alien species in Amazonian aquatic environments, thereby highlighting the TMPM as a critical hotspot in the Amazonian region.

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eDNA metabarcoding has been increasingly employed in the monitoring of marine invertebrate non-indigenous species (NIS), in particular using filtered seawater. However, comprehensive detection of all NIS may require a diversity of sampling substrates. To assess the effectiveness of 5 sample types (hard and artificial substrates, water, zooplankton) on the recovery of invertebrates' diversity, two marinas were monitored over three time points, using COI and 18S rRNA genes as DNA metabarcoding markers. We detected a total of 628 species and 23 NIS, with only up to 9% species and 17% of NIS detected by all sample types. Hard and artificial substrates were similar to each other but displayed the most significant difference in invertebrate recovery when compared to water eDNA and zooplankton. Five NIS are potential first records for Portugal. No NIS were detected in all sample types and seasons, highlighting the need for varied sampling approaches, and consideration of temporal variation for comprehensive marine NIS surveillance.

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Based on a search of publications in the scientific literature as well as international reports available online, I draw up a list of 25 documents which include cross-references to the terms offshore wind farms (OFW), and non-indigenous species (NIS). This review shows that no relationship has yet been clearly established between the implementation of OFWs and the colonization of NIS on turbine foundations and scour protections. Evidence for such an effect needs to be documented and confirmed in the future.

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Buttonweed (Cotula coronopifolia) is native to South Africa but invasive in wetlands in Europe, North America, and Australasia, where it excludes native plants. Despite being dry-fruited, field studies suggest migratory waterbirds can disperse its seeds via gut passage (endozoochory), aiding its expansion. To explore the potential for endozoochory in different regions and habitats, we collected seeds from six populations in Spain, Sweden, and the UK. Germination was tested under different salinity levels (0, 5, 10, 15 g/L) and simulated gut passage treatments: scarification, acidification, or both. No germination occurred at 15 g/L. Higher salinity reduced and delayed germination, but full gut passage treatment (i.e., both scarification and acidification) increased germinability and accelerated germination. Scarification or acid treatment alone resulted in intermediate germination patterns. There were significant salinity × population and gut passage × population interactions on germinability. The acceleration effect of gut passage on germination was stronger at 5-10 g/L than at 0 g/L. This study highlights how migratory birds can facilitate the spread of alien plants introduced by humans. Endozoochory by waterbirds is an understudied mechanism for the long-distance dispersal of dry-fruited alien plants. Further research on C. coronopifolia, including population genetics, is necessary to understand dispersal mechanisms and facilitate management strategies.

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In the past few years, non-native blue crab has been expanding its range in the Adriatic and Mediterranean Seas. However, when non-indigenous species establish a lasting presence in a novel ecosystem and actively expand into other regions, posing potential risks to local biodiversity and economic harm, they are classified as invasive species. For this study, two different batches of blue crabs were collected: one from the Mediterranean Sea and the other from the Adriatic coast area. Considering the negative ecological impact that blue crab has had and continues to have on the Italian coasts, this work was aimed to evaluate the characteristics of the Adriatic Sea "variant"; another goal was to propose the potential use of blue crab as a commercially profitable source due to its organoleptic characteristics. Data obtained revealed a high protein content, a good lipid profile, a low number of plastic particles, and the absence of toxic metals and pathogenic microorganisms. A deep statistical analysis was conducted to compare different portions of blue crab from the Adriatic and Mediterranean Seas. The results suggest that Callinectes sapidus is beneficial for human consumption and represents a valuable seafood, and its carapace could be used as compliant feed for aquaculture due to its metal composition and microbiological content.

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Despite the Baltic Sea being one of the most intensive shipping regions in the world the potential magnitude of the biofouled hulls in this region is unknown. This study estimated the biofouling load to Baltic Sea Region (BSR) based on the wetted surface area (WSA) method with regard to country, ship type and donor bioregion. WSA flux reached 656 km2, of which 86% was associated with ships operating inside and 14% was WSA flux brought by ships from outside of the Baltic Sea. Most of the WSA was transported to Swedish, Finnish and Danish ports as well. The highest WSA flux was assigned to roll-on/roll-off, passenger and general cargo ships. The high biofouling potential in BSR indicates a potential high risk to the environment and, therefore there is an urgent need for appropriate guidelines to be introduced into daily use by the commercial shipping community.

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Marine artificial structures provide substrates on which organisms can settle and grow. These structures facilitate establishment and spread of non-indigenous species, in part due to their distinct physical features (substrate material, movement, orientation) compared to natural habitat analogues such as rocky shores, and because following construction, they have abundant resources (space) for species to colonise. Despite the perceived importance of these habitat features, few studies have directly compared distributions of native and non-indigenous species or considered how functional identity and associated environmental preferences drive associations. We undertook a meta-analysis to investigate whether colonisation of native and non-indigenous species varies between artificial structures with features most closely resembling natural habitats (natural substrates, fixed structures, surfaces oriented upwards) and those least resembling natural habitats (artificial materials, floating structures, downfacing or vertical surfaces), or whether functional identity is the primary driver of differences. Analyses were done at global and more local (SE Australia) scales to investigate if patterns held regardless of scale. Our results suggest that functional group (i.e., algae, ascidians. barnacles, bryozoans, polychaetes) rather than species classification (i.e., native or non-indigenous) are the main drivers of differences in communities between different types of artificial structures. Specifically, there were differences in the abundance of ascidians, barnacles, and polychaetes between (1) upfacing and downfacing/vertical surfaces, and (2) floating and fixed substrates. When differences were detected, taxa were most abundant on features least resembling natural habitats. Results varied between global and SE Australian analyses, potentially due to reduced variability across studies in the SE Australian dataset. Thus, the functional group and associated preferences of the highest threat NIS in the area should be considered in design strategies (e.g., ecological engineering) to limit their establishment on newly built infrastructure.

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Plastic debris is a significant and rapidly developing ecological issue in coastal marine ecosystems, especially in areas where it accumulates. This study introduces "plasticlusters", a new form of floating debris agglomeration found in the Yasmine Hammamet marina (Tunisia, North-Africa), loosely attached to pontoon ropes around the water surface level. The analysis of two samples revealed that they were formed primarily by average 2.11 mm polystyrene fragments, 3.43 mm fibers, 104 mm polypropylene and polyethylene sheets, and 122 mm decomposing seagrass leaves. They were inhabited by several taxa, including at least 2 cryptogenic and 5 non-indigenous species (NIS). Unlike other plastic formations, plasticlusters provide a novel and potentially temporal microhabitat to fouling assemblages due to their loose and unconsolidated structure which, combined with marinas being NIS hubs, could enhance NIS dispersion. The results of this study raise concerns about the combined ecological effects of debris accumulation and biocontamination inside marinas.

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The recreational boating sector is a major vector for the introduction of non-indigenous species (NIS) via biofouling. Despite applying control measures to prevent the growth of fouling communities, most vessels are NIS carriers. This study assessed the effectiveness of different antifouling strategies in a manipulative experiment by testing two common coating typologies (biocide-based and foul-release coatings), accompanied with simulated maintenance practices. The experiment was carried out in the Gulf of La Spezia (Italy) and samples were collected at two different periods. Results showed significant differences among antifouling treatments regarding community structure, diversity, coverage and biovolume of the sessile component, alongside a significant decrease in the performance of biocide-based coating with time. Interestingly, peracarid NIS/native species ratio was higher for biocide-based treatments, suggesting potential biocide resistance. This study highlights the urgent need to develop common and feasible biofouling management plans and provides insights towards identification of best practices for recreational vessels.

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Molecular biomonitoring programs increasingly use environmental DNA (eDNA) for detecting targeted species such as marine non-indigenous species (NIS) or endangered species. However, the current molecular detection workflow is cumbersome and time-demanding, and thereby can hinder management efforts and restrict the "opportunity window" for rapid management responses. Here, we describe a direct droplet digital PCR (direct-ddPCR) approach to detect species-specific free-floating extra-cellular eDNA (free-eDNA) signals, i.e., detection of species-specific eDNA without the need for filtration or DNA extraction, with seawater samples. This first proof-of-concept aquarium study was conducted with three distinct marine species: the Mediterranean fanworm Sabella spallanzanii, the ascidian clubbed tunicate Styela clava, and the brown bryozoan Bugula neritina to evaluate the detectability of free-eDNA in seawater. The detectability of targeted free-eDNA was assessed by directly analysing aquarium marine water samples using an optimized species-specific ddPCR assay. The results demonstrated the consistent detection of S. spallanzanii and B. neritina free-eDNA when these organisms were present in high abundance. Once organisms were removed, the free-eDNA signal exponentially declined, noting that free-eDNA persisted between 24-72 h. Results indicate that organism biomass, specimen characteristics (e.g., stress and viability), and species-specific biological differences may influence free-eDNA detectability. This study represents the first step in assessing the feasibility of direct-ddPCR technology for the detection of marine species. Our results provide information that could aid in the development of new technology, such as a field development of ddPCR systems, which could allow for automated continuous monitoring of targeted marine species, enabling point-of-need detection and rapid management responses.

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Zooplankton are key components of estuarine trophic networks. However, routine monitoring is hindered by the difficulty of morphology-based identification. DNA-based methods allow us to circumvent some of these hurdles, providing precise species identifications regardless of the taxonomic expertise of the investigator or the developmental stage of the specimens. However, the process is dependent on the completeness of the reference libraries. In this study, we sought to evaluate the potential of DNA metabarcoding to assess the seasonal (summer, autumn, and early spring) and spatial dynamics of zooplankton (four locations spanning ca. 6 km) in the Lima estuary (NW Portugal). Two genetic markers were used: the cytochrome c oxidase subunit I and the V4 hypervariable region of the ribosomal 18S rRNA genes. Overall, 327 species were recovered, and both markers displayed minute overlap (7% were detected with both markers). Species richness, composition, and taxonomic distinctness were majorly influenced by the season, with a declining tendency from summer (highest number of exclusive species, n = 74) to spring. Second to season, the taxa composition was influenced by spatial variation where the most downstream site displayed the highest number of exclusive species, n = 53. A total of 16 non-indigenous species were detected using metabarcoding, but only one (Austrominus modestus) has been documented out in the estuary. In conclusion, both the seasonal and spatial gradients influenced the recovered richness, composition, and taxonomic distinctness, confirming the great aptitude of DNA metabarcoding for providing higher density monitoring and shedding new light on the composition and dynamics of complex zooplankton communities.

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This article documents, through a quantitative approach, the negative effect of the highly invasive species Amphistegina lobifera Larsen, 1976 on native benthic foraminiferal assemblages of coastal areas in the Sicily Channel (Central Mediterranean). A nested sampling design was applied through the comparison of benthic foraminiferal community structure across three areas that are known to be at different stages of invasion (i.e. Maltese Islands - advanced, southern Sicily - medium, and eastern Sicily - early). Results suggested that both diversity and richness of benthic foraminiferal community from the Maltese Islands were strongly modified by increased abundances of A. lobifera. In contrast, this phenomenon is less evident in southern and eastern Sicily, where the invader displayed lower abundances and the community structure was more diversified. Collected data also allowed for predicting what could happen in the near future in the whole Sicily Channel, as well as in the rest of the Mediterranean Sea.

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Recent arrivals of tar balls have been observed in several tropical beaches associated with the oceanic circulation that flows to the Brazilian continental shelf. Between August and September 2022, tar balls were collected in the northeastern coast of Brazil and analyzed. Nearly 90 % of the oils were colonized by barnacles, polychaetes, decapods, and algae. Most rafting organisms were Lepas anserifera with capitulum measuring 0.32 to 22.21 mm. Based on the growth rate of barnacles and the speed of the SEC it was estimated that tar balls were floating since July and August 2022 and traveled a maximum of 1938.82 km. The organisms and tar balls' possible origin is in the international waters, near to the meso-Atlantic ridge, known for oil tanker traffic. The tar balls, in addition to the oil-related impacts, can act as a vector of long-distance species dispersion, and it needs to raise an alert, considering the possible ecological impacts.

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An advanced characterization of the trophic niche of non-indigenous species (NIS) may provide useful information on their ecological impact on invaded communities. Here, we used carbon and nitrogen stable isotopes to estimate pairwise niche overlaps between non-indigenous and native consumers in the winter food web of Lake Trasimeno (central Italy). Overall, a relatively low pairwise overlap of isotopic niches was observed between NIS and native species. The only exception was the Louisiana crayfish Procambarus clarkii, which showed a relatively high and diffuse overlap with other native invertebrates. Our findings highlighted a high niche divergence between non-indigenous and native species in Lake Trasimeno, suggesting a potentially low degree of interspecific competition that may facilitate coexistence and, in turn, limit the strength of impacts. The divergent results obtained for the Louisiana crayfish indicate that additional control measures for this invasive species are needed to mitigate its impact on the Lake Trasimeno system.

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The introduction-rate of non-indigenous species (NIS) to coastal water bodies has accelerated over the last century. We present a model study assessing the fate of NIS released in likely point sources of the Danish Wadden Sea. We show that NIS-particles released in the deep North Sea are generally transported away from the Wadden Sea, while those released in the coastal North Sea and the Wadden Sea show large variability in track pattern and settlement location. Consequently, the introduction of NIS from ships entering the port of Esbjerg pose a threat to the Wadden Sea through primary and secondary spreading, while transport of species from sources in the south likely causes a slow and steady settling of NIS in the Wadden Sea and coastal North Sea. The study points to the importance of enforcing an efficient monitoring system to ensure early detection of changes to the species composition of the Wadden Sea.

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Biological invasions are a human-induced environmental disturbance that can cause major changes in ecosystem structure and functioning. Located in the northeastern Mediterranean basin, the Aegean Sea is a hotspot of biological invasions. Although the presence of alien species in the Aegean has been studied and monitored, no assessment has been conducted on their cumulative impacts on native biodiversity. To address this gap, we applied the CIMPAL index, a framework developed for mapping the cumulative impacts of invasive species, to identify the most affected areas and habitat types and determine the most invasive species in the region. Coastal areas showed stronger impacts than the open sea. The highest CIMPAL scores were four times more frequent in the South than in the North Aegean. Shallow (0-60 m) hard substrates were the most heavily impacted habitat type, followed by shallow soft substrates and seagrass meadows. We identified Caulerpa cylindracea, Lophocladia lallemandii, Siganus luridus, Siganus rivulatus, and Womersleyella setacea as the most impactful species across their range of occurrence in the Aegean but rankings varied depending on the habitat type and impact indicator applied. Our assessment can support marine managers in prioritizing decisions and actions to control biological invasions and mitigate their impacts.

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The shrimp Penaeus aztecus, native to the western Atlantic, was first reported in the eastern Mediterranean Sea in 2010. New records, from different Mediterranean localities, multiplied in the following years. The accurate search of the literature on non-indigenous species discovered it was misidentified more than once as another alien shrimp, P. semisulcatus, native to the Indo-Pacific region, with the result that its earlier presence in the Black Sea went unnoticed. Morphological characteristics allowing the identification of these two species, the autochthonous P. kerathurus and two other alien Penaeus species present in the Mediterranean, are reprised. The present distribution of P. aztecus based on literature records and surveys carried out in the northern and central Adriatic between 2016 and 2021 is mapped. The unintentional transport of larvae carried in ballast water by transoceanic vessels departing from the U.S. East Coast is suggested as the most probable introduction pathway. The significance of the correct identification of non-indigenous species, a "Descriptor" adopted in the Marine Strategy Framework Directive for determining the good environmental status of marine waters in the European States, is emphasized.

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In 2017, aquaculture producers of the Albufeira lagoon, Portugal, reported an invasion of tunicates that was disrupting mussel production, particularly the tunicate Styela plicata (Lesueur, 1823). A totally effective eradication method still does not exist, particularly for S. plicata, and the effects of the eradication treatments on bivalves' performance are also poorly understood. Our study examined the effectiveness of eradication treatments using three laboratory trials and five treatments (air exposure, freshwater immersion, sodium hypochlorite, hypersaline solution and acetic acid) for S. plicata, as well as their effects on survival and growth of blue mussel Mytilus edulis Linnaeus, 1758. While air exposure and freshwater immersion caused a 27% mortality rate in S. plicata, the acetic acid treatment was the most effective in eliminating this species (>90% mortality). However, a 33-40% mortality rate was registered in mussels. Both species were not affected by the hypersaline treatment in the last trial, but the sodium hypochlorite treatment led to a 57% mortality rate in mussels. Differences in mussels' growth rates were not detected. These trials represent a step forward in responding to the needs of aquaculture producers. However, further studies are needed to investigate the susceptibility of tunicates to treatments according to sexual maturation, as well as to ensure minimum mussel mortality in the most effective treatments, and to better understand the effects on mussel physiological performance in the long-term.

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The environmental similarity scores between source and recipient locations are essential in ballast water risk assessment (BWRA) models used to estimate the potential for non-indigenous species (NIS) introduction, survival, and establishment, and to guide management strategies aiming to minimize biodiversity loss and economic impacts. Previous BWRA models incorporate annual-scale environmental data, which may overlook seasonal variability. In this study, temporal variation in sea surface temperature and salinity data were examined at global ports, and the influence of this variation on environmental distance calculations (and corresponding risk of NIS) was examined for ballast water discharges in Canada by comparing outputs from monthly and annual scale assessments in a BWRA model. Except for some outliers in the Pacific region, the environmental distances based on monthly scale data generally become smaller in all regions, demonstrating that the model using annual decadal average environmental data to inform environmental matching can underestimate risk of NIS survival and establishment in comparison to monthly data. The results of this study suggest future evaluations incorporating the date of ballast water uptake and discharge can provide a more sensitive assessment of risk reflecting seasonal variability compared to an annual average risk model.

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Cassiopea andromeda (Forsskål, 1775), commonly found across the Indo-Pacific Ocean, the Red Sea, and now also in the warmest areas of the Mediterranean Sea, is a scyphozoan jellyfish that hosts autotrophic dinoflagellate symbionts (family Symbiodiniaceae). Besides supplying photosynthates to their host, these microalgae are known to produce bioactive compounds as long-chain unsaturated fatty acids, polyphenols, and pigments, including carotenoids, with antioxidant properties and other beneficial biological activities. By the present study, a fractionation method was applied on the hydroalcoholic extract from two main body parts (oral arms and umbrella) of the jellyfish holobiont to obtain an improved biochemical characterization of the obtained fractions from the two body parts. The composition of each fraction (i.e., proteins, phenols, fatty acids, and pigments) as well as the associated antioxidant activity were analyzed. The oral arms proved richer in zooxanthellae and pigments than the umbrella. The applied fractionation method was effective in separating pigments and fatty acids into a lipophilic fraction from proteins and pigment-protein complexes. Therefore, the C. andromeda-dinoflagellate holobiont might be considered as a promising natural source of multiple bioactive compounds produced through mixotrophic metabolism, which are of interest for a wide range of biotechnological applications.

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