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Este artículo contiene 10 páginas, 5 figuras, 1 tabla., The process of site selection and spatial planning has received scarce attention in the scientific literature dealing with marine restoration, suggesting the need to better address how spatial planning tools could guide restoration interventions. In this study, for the first time, the consequences of adopting different restoration targets and criteria on spatial restoration prioritization have been assessed at a regional scale, including the consideration of climate changes. We applied the decision-support tool Marxan, widely used in systematic conservation planning on Mediterranean macroalgal forests. The loss of this habitat has been largely documented, with limited evidences of natural recovery. Spatial priorities were identified under six planning scenarios, considering three main restoration targets to reflect the objectives of the EU Biodiversity Strategy for 2030. Results show that the number of suitable sites for restoration is very limited at basin scale, and targets are only achieved when the recovery of 10% of regressing and extinct macroalgal forests is planned. Increasing targets translates into including unsuitable areas for restoration in Marxan solutions, amplifying the risk of ineffective interventions. Our analysis supports macroalgal forests restoration and provides guiding principles and criteria to strengthen the effectiveness of restoration actions across habitats. The constraints in finding suitable areas for restoration are discussed, and recommendations to guide planning to support future restoration interventions are also included., This study was funded by the EASME–EMFF (Sustainable Blue Economy) Project AFRIMED (http://afrimed-project.eu/, grant agreement N. 789059), supported by the European Community.

To enrich spatio-temporal information on the distribution of alien, cryptogenic, and neonative species in the Mediterranean and the Black Sea, a collective effort by 173 marine scientists was made to provide unpublished records and make them open access to the scientific community. Through this effort, we collected and harmonized a dataset of 12, 649 records. It includes 247 taxa, of which 217 are Animalia, 25 Plantae and 5 Chromista, from 23 countries surrounding the Mediterranean and the Black Sea. Chordata was the most abundant taxonomic group, followed by Arthropoda, Mollusca, and Annelida. In terms of species records, Siganus luridus, Siganus rivulatus, Saurida lessepsianus, Pterois miles, Upeneus moluccensis, Charybdis (Archias) longicollis, and Caulerpa cylindracea were the most numerous. The temporal distribution of the records ranges from 1973 to 2022, with 44% of the records in 2020–2021. Lethrinus borbonicus is reported for the first time in the Mediterranean Sea, while Pomatoschistus quagga, Caulerpa cylindracea, Grateloupia turuturu, and Misophria pallida are first records for the Black Sea ; Kapraunia schneideri is recorded for the second time in the Mediterranean and for the first time in Israel ; Prionospio depauperata and Pseudonereis anomala are reported for the first time from the Sea of Marmara. Many first country records are also included, namely: Amathia verticillata (Montenegro), Ampithoe valida (Italy), Antithamnion amphigeneum (Greece), Clavelina oblonga (Tunisia and Slovenia), Dendostrea cf. folium (Syria), Epinephelus fasciatus (Tunisia), Ganonema farinosum (Montenegro), Macrorhynchia philippina (Tunisia), Marenzelleria neglecta (Romania), Paratapes textilis (Tunisia), and Botrylloides diegensis (Tunisia).

Habitat degradation and loss are severely affecting macroalgal forests worldwide, and their successful mitigation depends on the identification of the drivers of loss and the implementation of effective conservation and restoration actions. We made an extensive literature review 1- to document the historical (1789–1999) and recent (2000−2020) occurrence of the genus Cystoseira, Ericaria and Gongolaria reported in the literature along the 8000 km of the coasts of Italy, 2- to assess their decline and patterns of extinction, 3- to ascertain the drivers responsible for these changes, 4- to highlight the existence of success stories in their conservation and natural recovery. In the last twenty years, overall information on the distribution of Cystoseira s.l. exponentially increased, although research focused almost exclusively on intertidal reefs. Despite the lack of systematic monitoring programs, the local extinction of 371 populations of 19 different species of Cystoseira s.l. was documented across several regions, since 2000. Coastal engineering and poor quality of waters due to urban, agricultural or industrial activities were often documented as leading causes of habitat loss. However, the drivers of extinction were actually unknown for the majority of the populations and cause-effects relationships are scarcely documented. Although the proportion of protected populations increased to 77.8%, Marine Protected Areas are unlikely to guarantee adequate conservation efficacy, possibly also for the widespread lack of management and monitoring plans dealing specifically with Cystoseira s.l. species, and few evidences of natural recovery were observed. Our review shows the dramatic lack of baseline information for macroalgal forests, highlighting the urgent need for the monitoring of less accessible habitats, the collection of long-term data to unveil drivers of loss, and an updated reporting about the conservation status of the species of interest to plan future interventions.

This study aimed to assess the combined effects of ocean acidification (OA) and pollution to the polychaete Syllis prolifera inhabiting the CO

Fixatives are widespread in biological and medical research because they allow preserving specimens for a long time. Historically, formaldehyde has been the most used fixative so far, but new solutions are needed because of its carcinogenicity. In this study, we tested alternative fixative methods to find a harmless, economic, and simple-to-use methodology to fix samples for larval morphological analysis in Paracentrotus lividus. In two separate experiments, P. lividus embryos were fixed after 48 h post-fertilization by adding Formalin Free Tissue AccustainTM, NaOH-buffered Formalin Free Tissue AccustainTM, glacial ethanol and denatured ethanol at different concentrations (from 10 % to 70 %) and by submerging the vials containing the larvae in seawater at 0 °C and maintained at 4 °C for 144 h. Our results suggested that all the alternative fixatives tested do not guarantee a good quality of larvae for morphological purposes, while larvae that faced the thermal shock and were kept at 4 °C did not show any evidence of damage throughout time. The results of this study candidate this method as a good and safe substitute of formalin in studies that require morphological and taxonomic recognition and shed light on its use in other kinds of studies as well., Invertebrate Survival Journal, Vol 18 No 1 (2021)

In marinas and harbours, the accumulation of pollutants in sediments, combined with poor exchange of water with the open sea, poses a major environmental threat. The presence of photosynthetic organisms and the related oxygen production, however, may alleviate the negative effects of environmental contamination on heterotrophic organisms, enhancing their physiological defences. Furthermore, possible transgenerational buffer effects may increase the ability of natural populations to face environmental stress. Here we tested the occurrence of transgenerational effects on larvae of the sea urchin Paracentrotus lividus, whose parents were exposed, during the gametogenesis, to contaminated sediments subject to two temporal patterns of water re-suspension events and normal- (90%) vs. super-saturated (200%) levels of O2. The study site was Bagnoli-Coroglio (Gulf of Naples, southern Tyrrhenian Sea), a historically polluted brownfield and Site of National Interest for which environmental restoration options are currently under exploration. Larvae from different adult populations were significantly, although not linearly, affected by the interaction of all factors to which parents were exposed, at both 24h and 48h post fertilization. Specifically, the exposure of larvae to elutriates from contaminated sediments determined a developmental delay, a reduction in size and an increased percentage of abnormalities in all larval populations independently of their parental exposure. On the contrary, larvae from parents exposed to contaminated sediments, when reared in clean filtered sea water, succeeded in developing until the echinopluteus stage after 48h, with size and abundance comparable to those of larvae from control parents. Pre-exposure of parents to contaminated sediments did not successfully buffer the negative effects of elutriates on their offspring, and no positive effects of ‘super-saturated’ levels of O2 in response to contaminants were observed, suggesting that the Bagnoli-Coroglio area is currently not suitable for the re-stocking or re-introduction of this species.

A re-examination of marine alien species or Non Indigenous Species (NIS) reported in Italian Seas, until December 2018, is provided, focusing on establishment success, year of first record, origin, potential invasiveness, and likely pathways, in particular. Furthermore, their distribution is assessed according to the marine subregions outlined by the European Union (EU) Marine Strategy Framework Directive: Adriatic Sea (ADRIA), Ionian Sea and Central Mediterranean Sea (CMED), and Western Mediterranean Sea (WMED). In Italy, 265 NIS have been detected with the highest number of species being recorded in the CMED (154 species) and the WMED (151 species) subregions, followed by the ADRIA (143) subregion. Most of these species were recorded in more than one subregion. One hundred and eighty (180 or 68%) NIS have established stable populations in Italian Seas among which 26 have exhibited invasive traits. As regards the taxa involved, Macrophyta rank first with 65 taxa. Fifty-five of them are established in at least one subregion, mostly in the ADRIA and the CMED. Crustacea rank second with 48 taxa, followed by Polychaeta with 43 taxa, Mollusca with 29 taxa, and Fishes with 28 taxa, which were mainly reported from the CMED. In the period 2012-2017, 44 new alien species were recorded, resulting in approximately one new entry every two months. Approximately half of the NIS (~52%) recorded in Italy have most likely arrived through the transport-stowaway pathway related to shipping traffic (~28% as biofoulers, ~22% in ballast waters, and ~2% as hitchhikers). The second most common pathway is the unaided movement with currents (~19%), followed by the transport-contaminant on farmed shellfishes pathway (~18%). "Unaided" is the most common pathway for alien Fishes, especially in the CMED; escapes from confinement account for ~3% and release in nature for ~2%. The present NIS distribution hotspots for new introductions were defined at the first recipient area/location in Italy. In the ADRIA, the hotspot, Venice, accounts for the highest number of alien taxa introduced in Italy, with 50 newly recorded taxa. In the CMED subregion, the hotspots of introduction are the Taranto and Catania Gulfs, hosting 21 first records each. The Strait of Sicily represents a crossroad between alien taxa from the Atlantic Ocean and the Indo-Pacific area. In the WMED, bioinvasion hotspots include the Gulfs of Naples, Genoa and Livorno. This review can serve as an updated baseline for future coordination and harmonization of monitoring initiatives under international, EU and regional policies, for the compilation of new data from established monitoring programs, and for rapid assessment surveys.

The brown macroalgae of the genus Cystoseira are important habitat forming species along the rocky coasts all over the Mediterranean Sea. However, their decline at basin and local scale has been documented in many studies. We have characterized malacofauna associated with Cystoseira amentacea, C. compressa and C. crinita along the coasts of Ischia Island (Gulf of Naples). Samples were collected by snorkeling in the infralittoral belt. The surface within 20 x 20 cm frames was scraped off and collected in three replicates each sites. The diversity and structure of community were described by number of species, the exponential Shannon and the reciprocal Simpson’s indexes of diversity. The patterns of diversity at spatial scale were assessed by alpha, beta and gamma diversity. A total of 53 species of molluscs were identified in those associations. Gastropoda were the most species-rich class followed by Bivalvia and Polyplacophora. Bivalves were dominant in terms of number of individuals because of the mussel Mytilus galloprovincialis. The species M. galloprovincialis was the most frequent and top dominant one inhabiting Cystoseira associations along the coasts of Ischia Island (96.6 % of the total abundance). Most of the identified molluscs species belonged to two feeding guilds: micrograzers and filter feeders (29 and 13 species respectively). Only juveniles were found providing the importance of Cystoseira associations as nursery for molluscs recruitment. Differences in composition and structure of molluscs assemblages were found within the three algal associations and seem to correspond both to different morphology and habitat in which these algal species live.

The Mediterranean fruitflyCeratitis capitata(medfly) is an invasive agricultural pest of high economical impact and has become an emerging model for developing new genetic control strategies as alternative to insecticides. Here, we report the successful adaptation of CRISPR-Cas9-based gene disruption in the medfly by injectingin vitropre-assembled, solubilized Cas9 ribonucleoprotein complexes (RNPs) loaded with gene-specific sgRNAs into early embryos. When targeting the eye pigmentation genewhite eye(we), we observed a high rate of somatic mosaicism in surviving G0 adults. Germline transmission of mutatedwealleles by G0 animals was on average above 70%, with individual cases achieving a transmission rate of nearly 100%. We further recovered large deletions in thewegene when two sites were simultaneously targeted by two sgRNAs. CRISPR-Cas9 targeting of theCeratitisortholog of theDrosophilasegmentationpairedgene (Ccprd) caused segmental malformations in late embryos and in hatched larvae. Mutant phenotypes correlate with repair by non-homologous end joining (NHEJ) lesions in the two targeted genes. This simple and highly effective Cas9 RNP-based gene editing to introduce mutations inCeratitis capitatawill significantly advance the design and development of new effective strategies for pest control management.

The Mediterranean fruitfly Ceratitis capitata (medfly) is an invasive agricultural pest of high economic impact and has become an emerging model for developing new genetic control strategies as an alternative to insecticides. Here, we report the successful adaptation of CRISPR-Cas9-based gene disruption in the medfly by injecting in vitro pre-assembled, solubilized Cas9 ribonucleoprotein complexes (RNPs) loaded with gene-specific single guide RNAs (sgRNA) into early embryos. When targeting the eye pigmentation gene white eye (we), a high rate of somatic mosaicism in surviving G0 adults was observed. Germline transmission rate of mutated we alleles by G0 animals was on average above 52%, with individual cases achieving nearly 100%. We further recovered large deletions in the we gene when two sites were simultaneously targeted by two sgRNAs. CRISPR-Cas9 targeting of the Ceratitis ortholog of the Drosophila segmentation paired gene (Ccprd) caused segmental malformations in late embryos and in hatched larvae. Mutant phenotypes correlate with repair by non-homologous end-joining (NHEJ) lesions in the two targeted genes. This simple and highly effective Cas9 RNP-based gene editing to introduce mutations in C. capitata will significantly advance the design and development of new effective strategies for pest control management.

A sulfated polysaccharide was successfully isolated from Cymodocea nodosa (CNSP). This is the first report that indicates the chemical composition, structural characterization, functional and antihypertensive properties of this polysaccharide. The CNSP consisted mainly of sulfate (23.17%), total sugars (54.90%), galactose (44.89%), mannose (17.30%), arabinose (12.05%), xylose (9.18%), maltose (1.07%) and uronic acid (11.03%) with low water activity (0.49). CNSP had an XRD pattern that was typical for a semi-crystalline polymer with homogeneous structure. It also displayed an important anti-hypertensive activity (IC50 = 0.43 mg ml) with a dose-dependent manner using a synthetic substrate, N-hippuryl-His-Leu hydrate salt (HHL). Overall, the results indicate that CNSP have attractive chemical, functional and biological properties, with a preliminary structural may have a backbone of branched 6-O-sulfated (1 → 4) galactosidic linkages, which can be considered in the future as alternative additive in various foods, cosmetic and pharmaceutical preparations.

Brown macroalgae belonging to the genus Cystoseira (Fucales: Sargassaceae) are canopy-forming organisms the recent decline of which at a basin and local scale has been widely documented, which urgently calls for research to fill knowledge gaps and support new and effective measures for protection. We, hereby, characterised the molluscan assemblages associated with three Cystoseira taxa (C. amentacea, C. compressa and C. crinita) from Ischia Island (Italy, Tyrrhenian Sea), and tested whether different congeneric taxa may syntopically support a different biota. In particular, these assemblages were compared among three Cystoseira species, between two times of sampling (June–July 2015 and June–July 2016), and among six sites in terms of multivariate structure (identity and relative abundances of constituting taxa combined, and presence–absence composition), as well as for synthetic measures of diversity, including the total richness of taxa, the exponential Shannon index and the reciprocal Simpson index. In total, 24736 molluscan individuals were collected, overall belonging to 52 taxa. The majority of the identified species included micrograzers and filter feeders, which is in agreement with similar previous studies. The composition of associated molluscan assemblages, which was mainly represented by juvenile individuals, differed among the three Cystoseira species, suggesting that even congeneric taxa do not support an analogous benthic fauna. The present findings have shed light on the molluscan biota associated with Cystoseira taxa in the Gulf of Naples and strengthened the importance of such habitat-forming macroalgae in structuring the local infralittoral invertebrate biodiversity and as a nursery for species-specific associated molluscs.