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In the Mediterranean, Cystoseira sensu lato (s.l.) (Phaeophyceae) forests have sharply declined and restoration measures are needed to compensate for the loss. Assisted regeneration through the outplanting of seedlings grown ex-situ has proven to be a sustainable option. Optimizing mesocosm culture can maximize survival of the most critical embryonic stages and reduce long-term maintenance costs. Host-microbiome interactions could also play a crucial role in seedling development and welfare. In this context, we aimed to advance a cultivation protocol that stimulates the growth and fitness of Ericaria amentacea (Phaeophyceae) seedlings and identify the associated microbial biofilm communities. Seedlings were cultured in 6 treatments [i.e., filtered seawater (SW, C, Control), von Stoch-enriched SW (VS), VS + algal extract (VSA), algal extract-enriched SW: A1< A2< A3]. After the field, A2 seedlings had the highest cover (1372 ± 53.66 mm2), which was 1.8 and 1.9 times greater than in VS and VSA, respectively. The addition of the algal extract and nutrients significantly affected the structure and composition of the microbial community that shifted over time in each culture medium. We identified a treatment-specific microbial fingerprint. After the mesocosm phase, A2 was characterized by 4 unique taxa: Postechiella, Winogradskyella, Roseovarius and Arenibacter (Bacteria). Given the success of A2 seedlings, we propose the probiotic consortia candidates characterized by the unique treatment-taxa in conjunction with the shared taxon Psychroserpens (Bacteria, present in A1, A2, VSA, VS) and the reminder community. Within the holobiont concept, the effect of algal extract or nutrients on the algae and/or biofilm could have important consequences for tuning the overall interaction networks. Our study has shown that macroalgal restoration could benefit from both the use of commercial algal extract and tailored nutrient enrichment in ex-situ cultures and the identification of probiotic consortia candidates that promote seedling growth.

Several anthropogenic factors are responsible for the decline of Cystoseira sensu lato (hereafter Cystoseira) forests along Mediterranean coasts. Some Cystoseira species are already regionally extinct, and their decline has been widely recorded. Sustainable and efficient techniques for the restoration of Cystoseira are needed. In this context, the objectives of this study were i) to analyse the reproductive traits of three populations of Gongolaria barbata from three nearby donor sites in the northern Adriatic Sea, assessing the differences in their reproductive potential and reproductive success; and ii) to evaluate the outplanting success in terms of the effectiveness of G. barbata restoration, in relation to the different donor and receiving sites (Miramare MPA and in front of the Marine Biology Station Piran - MBSP) and implemented methods (ex situ and hybrid method combining a mesocosm cultivation and a suspended culture in the field). After 2 weeks of cultivation in mesocosms, half of the tiles with germlings were transported to the receiving sites and placed on suspended lantern nets (hybrid method), which were later (after 3 months) transferred to the seabed on concrete plates with protective cages. The remaining tiles were placed on the seabed on concrete plates with protective cages after a 4-week culture (ex situ method). At both sites, lantern nets and plates were randomly placed at 3 m depth. Thallus length was measured monthly in each treatment. Seedlings in suspended culture showed lower performance at the Miramare MPA, most likely due to the unfavourable environmental conditions. The satisfactory results obtained at MBSP demonstrate the efficiency of the hybrid method and confirm its potential to reduce the cost and time required for cultivation. Since unpredictable climatic events pose the greatest threat to restoration performance, these challenges must be considered when establishing new restoration practices. Moreover, herbivore regulation is extremely urgent and should be planned and implemented on a larger regional scale.

Canopy-forming brown algae support highly productive ecosystems whose decline has been attributed to the interplay of several anthropogenic disturbances. Climate change could have disruptive effects on the biology of these species, but the role of temperature in the development of early life stages is poorly understood. The aim of this study was to assess the response of Ericaria giacconei, a winter-reproducing Southern–Mediterranean endemic species, to thermal stress by testing five temperatures (12, 15, 18, 24, and 28°C) on adults and early stages. Chlorophyll a fluorescence of adult plants was measured at 0, 24, 72, and 120 h on nine fronds in each of the three aquaria per treatment. To assess egg release, zygote settlement, and embryo growth rate, approximately 1,200 receptacles were cultured on six Petri dishes per temperature treatment, and 10 random subsections of 2 ×2 mm were examined in three Petri dishes at 0, 20, 44, and 92 h after fertilization. Adult plants showed a plastic physiological response, and thermal stress had no significant effect on PSII efficiency. Embryos fully developed only at 12 and 15°C. Mortality increased at 18 and 24°C, and no zygotes survived at 28°C. In a scenario of further increasing temperatures, the effects of warming could affect the recruitment of E. giacconei and increase its vulnerability to further stresses. These effects on the survival of early stages, which are the bottleneck for the long-term survival of the species, should be taken into account in conservation and restoration measures to maintain canopy-forming macroalgal populations and associated biodiversity and ecosystem services.

Marine Protected Areas (MPAs) are at the core of management and conservation plans aimed at counteracting the ongoing widespread degradation of marine ecosystems. Although the outcomes of protection in enhancing marine biodiversity and restoring populations of exploited species are widely recognized, the putative effect of MPAs in increasing the resilience of marine communities and ecosystems is still underexplored. In this study, long-term data from a no-take area of a Mediterranean MPA and unprotected locations were used to investigate the effects of full protection on the temporal stability of sessile assemblages on shallow subtidal rocky reefs. Results highlighted that assemblages within the no-take area exhibited a more consistent structure over 20 years of study, whereas a significantly larger temporal variability characterized assemblages at the unprotected locations. Reduced levels of human pressure and stabilizing processes related to the re-establishment of the integrity of trophic networks within the MPA could underlie the observed patterns. Assessing whether MPAs may improve the ability of marine communities to absorb disturbance, from local to large scale sources, is essential to ascertain their contribution to future adaptive conservation strategies in the face of global change. This study provided evidence that MPAs, if well managed, can represent an effective tool to smooth fluctuations in the structure of marine communities, thus potentially reducing their vulnerability to natural and human-driven environmental changes.

High complexity habitat traits (i.e. high heterogeneity and/or size of discrete habitat units) often promote larger abundances of fauna. Sandy and rhodolith sea bottoms are typically interspersed as mosaics within coastal landscapes. The aim of this study was to experimentally assess the effect of two complexity attributes of rhodolith nodules (i.e. their heterogeneity and size) on the abundance and structure of vagile epibenthic assemblages. An early-colonization experiment was set up (July 2016), where experimental units containing rhodolith nodules of varying heterogeneity and size were deployed at two adjacent recipient habitats: a sandy bottom and a rhodolith seabed. After one month, the abundance of fauna colonizing the experimental units was similar in both habitats, but the assemblage structure (i.e. composition) notably differed. Importantly, the heterogeneity, rather than the size, of experimental rhodolith nodules influenced patterns of vagile macrofauna (>0.5 mm) colonization, despite the habitat type where these experimental units were deployed considerably influenced colonization patterns. This result reinforces the idea of the importance of rhodolith tri-dimensional structure, as key influencer on faunal communities of nearshore habitats.

Rhodolith beds (RBs) are bioconstructions characterized by coralline algae, which provide habitat for several associated species. Mediterranean RBs are usually located in the mesophotic zone (below 40 m), and thus are frequently remote and unexplored. Recently, the importance and vulnerability of these habitats have been recognized by the European Community and more attention has been drawn to their investigation and conservation. This study reports the results of an extensive monitoring program, carried out within the Marine Strategy Framework Directive (2008/56/EC), in six sites off the Campania coast (Italy, Mediterranean Sea). New insights were given into the distribution, cover, vitality (i.e., live/dead rhodolith ratio), structural complexity, and coralline algae composition of RBs. Remotely operated vehicles (ROV) investigations allowed the description of several RBs, and the discovery of a RB with rhodolith cover &gt, 65% offshore the Capri Island. Only two sites (Secchitiello and Punta Campanella) showed a very low mean cover of live rhodoliths (&lt, 10%), hence, not being classifiable as RBs. The collected rhodoliths were mostly small pralines (~2 cm), spheroidal to ellipsoidal, with growth-forms ranging from encrusting/warty to fruticose/lumpy. Coralline algae identification revealed a high diversity within each bed, with a total of 13 identified taxa. The genus Lithothamnion dominated all sites, and Phymatolithon calcareum and Lithothamnion corallioides, protected by the Habitats Directive (92/43/EEC), were detected in all RBs.

Macroalgal forests are one of the most productive and valuable marine ecosystems, but yet strongly exposed to fragmentation and loss. Detailed large-scale information on their distribution is largely lacking, hindering conservation initiatives. In this study, a systematic effort to combine spatial data on Cystoseira C. Agardh canopies (Fucales, Phaeophyta) was carried out to develop a Habitat Suitability Model (HSM) at Mediterranean scale, providing critical tools to improve site prioritization for their management, restoration and protection. A georeferenced database on the occurrence of 20 Cystoseira species was produced collecting all the available information from published and grey literature, web data portals and co-authors personal data. Data were associated to 55 predictor variable layers in the (ASCII) raster format and were used in order to develop the HSM by means of a Random Forest, a very effective Machine Learning technique. Knowledge about the distribution of Cystoseira canopies was available for about the 14% of the Mediterranean coastline. Absence data were available only for the 2% of the basin. Despite these gaps, our HSM showed high accuracy levels in reproducing Cystoseira distribution so that the first continuous maps of the habitat across the entire basin was produced. Misclassification errors mainly occurred in the eastern and southern part of the basin, where large gaps of knowledge emerged. The most relevant drivers were the geomorphological ones, followed by anthropogenic variables proxies of pollution and urbanization. Our model shows the importance of data sharing to combine a large number of spatial and environmental data, allowing to individuate areas with high probability of Cystoseira occurrence as suitable for its presence. This approach encourages the use of this modelling tool for the prediction of Cystoseira distribution and for supporting and planning conservation and management initiatives. The step forward is to refine the spatial information of presence-absence data about Cystoseira canopies and of environmental predictors in order to address species-specific assessments.

Rhodolith beds represent a key habitat worldwide, from tropical to polar ecosystems. Despite this habitat is considered a hotspot of biodiversity, providing a suite of ecosystem goods and services, still scarce quantitative information is available thus far about rhodolith beds occurrence and ecological role, especially in the Mediterranean Sea. This study reports the composition and patterns of distribution of rhodolith assemblages found in four study areas along ca. 860 km of coast in the Central Mediterranean Sea. These rhodolith beds were studied for the first time and significant differences at all spatial scales have been highlighted, documenting the high variability of this habitat. Rhodolith species composition, morphology and distribution have been discussed considering the potential role of environmental factors in driving these patterns. The need for improving their protection is discussed to complement present conservation and management initiatives, particularly in the frame of the EU Marine Strategy Framework Directive.

Connectivity is a fundamental ecological property affecting stability, resilience and recovery of marine populations, in particular in networks of patchy habitats as the Mesophotic Biogenic Habitats of the Northern Adriatic Sea. Specific information on the dispersal behaviour of many species living in these habitats is lacking, thus the connectivity simulations were performed over different pelagic propagules duration and different reproductive seasons of six consecutive years, covering most of the possible dispersal behaviour variability. The connectivity simulations were analysed with graph analysis methods by characterizing each connectivity graph with several structural measures and by identifying sites with greatest individual and group centrality. We tested the hypothesis that geographic proximity of sites cannot be a proxy for the connectivity and that hydrodynamic connectivity is statistically significantly related to the observed beta diversity among the studied sites. Results showed that species with different pelagic propagules duration follow greatly different dispersal dynamics, and that the network of studied outcrops is only partially connected to coastal benthic populations. Our results also made possible a critical appraisal of the current conservation strategies, evidencing gaps in the existing network of protected Mesophotic Biogenic Habitats in the Northern Adriatic Sea. We suggest possible improvements for an efficient preservation strategy of these unique biodiversity hotspots by including a fundamental ecological process, the dispersal connectivity, into the process of decision making for conservation.

Structure and spatial variability of coralligenous assemblages of the Apulian continental shelf in the southern Adriatic Sea were investigated. The area consists of small coralligenous outcrops distributed between 30 and 100 m of depth on coarse detritic or muddy bottom. Photographic samples, obtained through a Remotely Operated Vehicle, were analyzed to evaluate the abundance of the main taxa or morphological groups of macroalgae and sessile invertebrates, and the deposited sediment. A multifactorial sampling design was used to compare assemblages at two depth ranges, the shallow outcrops (between 35 and 45 m) and the deep outcrops (between 60 and 70 m), and to determine the main scales of spatial variation. Assemblages were dominated by encrusting and erect sponges, encrusting and filamentous algae, scleractinians, encrusting and erect bryozoans and the Zoantharia Parazoanthus axinellae. Deep assemblages were characterized by lower alpha and beta diversity, and by the decrease or disappearance of sensitive organisms, such as coralline macroalgae, scleractinians and erect bryozoans, and by the dominance of encrusting sponges, P. axinellae and hydrozoans. Sedimentation was higher on deep outcrops and was suggested to be the main driver of differences between shallow and deep assemblages. The peculiar composition of assemblages found in the Apulian continental shelf provides evidence that these outcrops represent an uncommon coralligenous habitat.

One of the objectives of the BALMAS project was to conduct Port Baseline Biological Surveys of native and non-indigenous benthic flora in 12 Adriatic ports. Samples of macroalgae growing on vertical artificial substrates were collected in spring and autumn 2014 and/or 2015. A total number of 248 taxa, 152 Rhodophyta, 62 Chlorophyta, and 34 Ochrophyta, were identified. Of these, 13 were non-indigenous seaweeds, mainly filamentous macroalgae, that were probably introduced through hull fouling. Some of these taxa had already been described in the study areas, others were recorded for the first time, a few were no longer detected at sites where they had previously been recorded (e.g. Sargassum muticum). Some other NISS reported for the Adriatic Sea, were not collected at any sampling site (i.e. Caulerpa cylindracea, Codium fragile). Possible reasons for the absence of these species are discussed.

This article gives an up-to-date review of the status of and main changes in benthic algal flora that have occurred in recent decades along the Italian Adriatic coastline. Common traits among the main structural/functional changes observed and their causes are discussed. A synthesis of the challenges to and prospects of filling gaps in the data, ecological knowledge and protection measures are also given.

In the Mediterranean region Cymodocea nodosa is widely distributed throughout shallow sites. Therefore, a correct assessment of the status of its meadows is of great importance for the implementation of the Water Framework Directive (WFD), the Marine Strategy Framework Directive (MSFD), and the Habitat Directive (HD), especially for areas where Posidonia oceanica meadows (the most frequently used indicator in the Mediterranean Sea) are rare or not present. The previously proposed index for the evaluation of the status of C. nodosa meadows (CymoSkew) is based on ln-transformed relative frequencies of photosynthetic leaf lengths, which we believe is statistically questionable. Therefore, we further improved the methodology and developed a new index named MediSkew, where ln-transformation is applied to raw data of leaf lengths. More specifically, the index is a combination of two metrics, both based on C. nodosa leaf length: deviation from the reference median length (Medi-) and skewness of the length frequency distribution (-Skew), though greater importance was assigned to the first. To develop the classification criteria for the assessment of the status, also a Pressure Index for Seagrass Meadows (PISM), for the evaluation of pressureeimpact relationships, was developed. The MediSkew is meant to be a rapid screening method for wide areas, therefore the index should be tested for the assessment of the status of C. nodosa meadows throughout Mediterranean coastal waters.

To evaluate the effect of grazing by sea urchins on the development of vegetation, plastic netting (6545-mm mesh) was stretched over the horizontal surfaces of two out of four cubes of a Loano artificial reef pyramid and held slightly above the surface. The macroalgal population of the horizontal surfaces of the four cubes was seasonally sampled over one and a half years. A qualitative analysis based on a floristic survey of all species present did not reveal significant differences between treatments. However, the quantitative analysis of the algal community based on biomass samples revealed marked differences. Before starting the experiment, fresh and dry weights of the vegetation of all four cubes were of comparable magnitude, while after deployment of the netting, biomass on the two surfaces protected by the net was always significantly higher than in the controls throughout the sampling period. The relative increase seems related to the mechanical effect of the netting in reducing the predatory pressure caused by sea urchins. 2002 International Council for the Exploration of the Sea. Published by Elsevier Science Ltd.

Publisher Summary This chapter presents a multidisciplinary study of the major characteristics of a set of submarine rock outcrops in the Northern Adriatic Sea: the rock occurrence, embedded down to about 1 m from the seafloor in the bottom sediments; numerous gas accumulations in the proximity of the outcrops; small-scale mud volcanoes; and microbial mats around the seepage site. From the biological point of view, a total of 112 macroalgal taxa were recorded. The different number of taxa recorded at the San Pietro and Bardelli sampling sites have been related to the different distance from the coast and to water depth. The data highlights that the seabed is almost flat, locally showing the occurrence of rock outcrops, with a range of sizes, shapes, and spatial orientations. On the side-scan survey (SSS) data, they appear as high backscatter features. The seabed is mainly composed of sands. Some species that characterize these outcrops are acknowledged as important bioconstructors distinctive of the Mediterranean area. The seafloor sedimentary deposits show a gradual increase of the fine fraction from the coast toward the offshore; then a progressive increase in the coarse fraction in the central part of the study area (residual sands of the TST) is recognized. The fragile biological equilibrium of these structures, the slow-growing calcareous organisms considered as nonrenewable resources, and the dredging and bottom trawling pressure that strongly threatens the physical characteristics of the habitat, and negatively affect the associated biota, all motivate the need to protect the Trezze and their surrounding seabed.