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Pollution associated to marine plastic litter is raising increasing concerns due to its potential harmful effects on human health, biota, and coastal ecosystems. However, limited information is available on the degradation behavior of plastics, especially biodegradable ones, in dune habitats. Moreover, the effects of plastics on dune plant growth and ability to withstand environmental stresses and invasion by non-native plants have been largely neglected. This is a particularly relevant issue since biological invasions are major threats to dune ecosystems. In this 18-month study, we examined the degradation behavior of two plastic bags, non-biodegradable (NBP) or biodegradable/compostable (BP), in the dune environment by visual observations and analytical techniques. Concomitantly, we investigated the individual and combined effects of bag type and sand burial (no burial vs. partial burial) on the performance of a native dune plant (Thinopyrum junceum) and an invasive plant (Carpobrotus sp.) and on their interaction. NBP did not show relevant degradation signs over the experimental period as expected. BP exhibited gradual surface modifications and changes in chemical functionality and were almost disintegrated after 18 months. Bags and burial reduced independently T. junceum survival and growth, and most plants died within 8 months of plastic exposure. Bags and burial did not affect Carpobrotus survival. However, burial decreased Carpobrotus growth while NBP increased it. Both plastics increased Carpobrotus competitive ability, and no T. junceum plants survived to co-occurrent Carpobrotus, BP, and burial. These findings indicate that removing all littered plastics from beach-dune systems not only is critical to reduce plastic pollution but also to prevent further spread of invasive species in coastal dunes.

Beach litter can affect public health and economic activities worldwide forcing local authorities to expensive beach cleaning. Understanding the key mechanisms affecting the accumulation of this waste on beaches, such as sea state and proximity to entry points, is critical to plan effective management strategies. In this one-year study, we estimated the impact of storm events and waterways runoff on litter abundance and local economy using as a model a managed, peri-urban beach facing a north-western sector of the Mediterranean Sea. We also investigated the relationship between litter composition/density and beach proximity to major/closest harbors/rivers at regional scale by combining our data with those on litter density available in literature. Autumn/winter storms caused larger litter depositions than spring/summer ones in the peri-urban beach. No preferential accumulation occurred near to waterway mouths. Litter mainly consisted of plastic, and its composition in terms of micro-categories varied over seasons. In total, 367,070 items were deposited along 4.7 km of beach over one year, and the cost for the removal of this waste amounted to approximately 27,600 euros per km/year. At regional scale, beach litter density was positively correlated to the proximity of major harbors while its composition was related to the proximity to both major harbors and rivers. Results indicate that autumn/winter storms are important drivers of marine litter deposition. They also suggest that beaches in front of the convergence zone of littoral currents and close to major harbors can be particularly subjected to this kind of pollutant. To increase their effectiveness, litter mitigation/cleaning activities should be planned based on predictions of major storm events and performed at spatial scales encompassing at least coastal regional sectors.

Plants play a fundamental role in maintaining coastal dunes but also accumulate littered microplastics (MPs). Migration tests suggest that naturally weathered MPs can leach out a broader range of potentially phytotoxic chemicals than virgin MPs. Thus, assessing MPs effects on plants using beached-collected particles rather than virgin ones is critically important. Here, the effects on plants of leachates from two pools of beach-collected and virgin MPs, high-density polyethylene (HDPE) and polypropylene (PP), and their mixture, were explored combining toxicity tests and chemical analyses. Phytotoxicity of MP leachates at different dilutions was evaluated under standard laboratory conditions using test species and under environmentally realistic conditions using the dune species Thinopyrum junceum. Leachates from beached PP and HDPE adversely affected all species, and the extent of these effects varied according to polymer type, concentration, and species. Virgin MPs had weaker effects than beached ones. Several potentially phytotoxic oxidized compounds were detected in water by GC/MS analysis, and their amount estimated. Results indicate that the molecular species leaching from beached MPs - at ppm concentration levels for the individual chemical species - can inhibit plant growth, and the effects of leachates from mixtures of degraded MPs can differ from those from individual polymers, highlighting the need for further investigation of MPs consequences for coastal ecosystems.

To assess the exposure of beachgoers to viruses, a study on seawater, sand, and beach-stranded material was carried out, searching for human viruses, fecal indicator organisms, and total fungi. Moreover, for the first time, the genome persistence and infectivity of two model viruses was studied in laboratory-spiked sand and seawater samples during a one-week experiment. Viral genome was detected in 13.6 % of the environmental samples, but it was not infectious (Human Adenovirus - HAdV, and enterovirus). Norovirus and SARS-CoV-2 were not detected. The most contaminated samples were from sand and close to riverine discharges. In lab-scale experiments, the infectivity of HAdV5 decreased by ~1.5-Log

A large fraction of plastic litter found in natural environments is constituted by conventional not biodegradable plastic bags, and their adverse effects via ingestion or entanglement on terrestrial and marine organisms are largely documented. Biodegradable and compostable shoppers have been recently developed as alternative to traditional ones. These bags are specifically designed to degrade in composting facilities and generate a product devoid of toxicity to soils and crops. However, very little is known on the effects of bag leaching, i.e. the transfer of chemicals from plastic into natural environments, on vegetation. Some plant species are highly sensitive to a variety of chemicals, and seedling growth is generally the most affected life history stage. In this study we assessed the potential effects of conventional (high-density polyethylene, HDPE) and compostable (Mater-bi®, MB) bags, when left in natural environments, on water quality and plant development. To this end, seeds of Lepidium sativum L., a terrestrial plant commonly used in phytotoxicity standard tests, were exposed to leachates obtained from different amount of HDPE and MB bags, simulating various pollution degrees occurring in nature, for 72 h. Both not-exposed (or virgin) bags and natural weathering exposed bags were used. Variations of chemical-physical characteristics of extracts were used as indicative of water quality deterioration, while alterations of seed gemination and seedling radicle and hypocotyl length were considered as indicative of phytotoxicity. A chemical qualitative screening of the leachates was also performed to identify the compounds with potential phytotoxicity. Both types of bags affected water characteristics (pH, salinity and total dissolved solids) relevant to plants, and released into water intentionally added chemicals, such as the noxious bisphenol A, and other phytotoxic substances probably generated during bag manufacturing. Leachates from both bag types did not affect seed germination. But, a significant number of seedlings showed developmental abnormalities or reduced seedling growth. The hypocotyl was the most sensible seedling organ to HDPE bag leachates while the radicle was the most vulnerable to MB ones. These findings indicate that plastic bags, including those that meet biodegradability and compostability standards, represent a potential threat to plants, if left in natural environments. Therefore, people and managers should be adequately informed about the potential environmental impact of an incorrect bag disposal. Simple, rapid standard phytotoxicity tests, such as the L. sativum bioassay, applied to bag leachates could be used in the future to select not noxious additives so to develop more eco-friendly bags.

Plastics are found in marine environments worldwide, and their effects on macrophytes (seagrasses and macroalgae) colonizing sandy bottoms are still poorly known. Seagrass meadows are valuable but declining ecosystems due to local and global-change related stressors, including sediment disturbance and introduced macroalgae. Understanding whether plastics pose a further threat to seagrasses is critically important. In two simultaneous additive experiments performed in an aquaculture tank, we examined the individual and combined effects of macroplastics (non-biodegradable high-density polyethylene and biodegradable starch-based) and sedimentation (no and repeated sedimentation) on the performance (in terms of biomass and architectural variables) of a native Mediterranean seagrass (Cymodocea nodosa) and an introduced macroalga (Caulerpa cylindracea), and on the intensity of their interactions. Macroplastics were still present in sediments after 18 months. Cymodocea nodosa produced a greater biomass and longer horizontal rhizome internodes forming clones with more spaced shoots probably to escape from plastics. Plastics prevented C. nodosa to react to sedimentation by increasing vertical rhizome growth. Under C. cylindracea invasion, C. nodosa allocated more biomass to roots, particularly to fine roots. In the presence of C. nodosa, C. cylindracea performance was reduced. High-density polyethylene (HDPE) plastic and sedimentation shifted species interactions from competitive to neutral. These results suggest that both HDPE and biodegradable starch-based macroplastics, if deposited on marine bottoms, could make seagrasses vulnerable to sedimentation and reduce plant cover within meadows. HDPE plastic and sedimentation could contribute to the decline of seagrass habitats by facilitating the spread of non-native macroalgae within meadows. Overall, the study highlights the urgent need to implement more effective post-marketing management actions to prevent a further entering of plastics in natural environments in the future, as well as to establish to conservation measures specifically tailored to protect seagrass habitats from plastic pollution.

Marine litter, that is solid materials deliberately or accidentally released into marine and coastal environments, is a growing issue both at local and global scale. In particular, litter accumulation on beaches represents a relevant problem in touristic coastal areas. The DPSIR framework, already applied to tackle other environmental issues, can help in analysing the origin, impact, and possible solution of material stranded on the beach allowing a holistic approach in the “one health” perspective. It was applied in a popular sandy bathing location in the North-Western Tuscany (Italy). Information and data on origin and fate of beach-stranded material as well as existing local strategies were retrieved from a stakeholders’ network. In summer 2020, a monitoring campaign was performed to evaluate the sanitary state of the shoreline and the impact of this material on coastal habitat. In a 7-year period (2014-2020), a mean amount of 1.548,3 tons of beach-stranded material have been collected and treated to separate sand (55%), organic green fraction (31%), and anthropogenic debris (2%), while 12% weight-loss was attributable to evaporation. Analysis of historical environmental data revealed that shoreline accumulation of this material was associated with low atmospheric pressure, medium-strong winds, and storms. Preliminary monitoring data showed that the green fraction was mainly represented by wood and terrestrial plants with little quantity of the seagrass Posidonia oceanica, while most of the anthropogenic debris was rubber and metal. Microbial pollution of beached material was at least one order of magnitude higher than the surrounding environment (sand and seawater) for total coliforms, Escherichia coli, intestinal enterococci, and total fungal load. Overall, our results indicate a potential impact of beach-stranded material in this coastal area, with possible health risk for human population. The instruments already in place for the management of beached material, although consistent, could be further improved.

Following the “Letter to the Editor regarding the article “Evaluation of the phytotoxicity of conventional and biodegradable plastic bags using seed germination tests” by Balestri et al. (2019) published on Ecological Indicators 102(2019):569–580” by Degli-Innocenti et al. (2019), we replied to the concerns expressed by Degli Innocenti and co-authors in this letter regarding our article.

Marine plastic litter is a global concern. Carrier bags manufactured from non-biodegradable polymers constitute a large component of this litter. Because of their adverse impact on marine life, non-biodegradable bags have recently been replaced by biodegradable ones. However, growing evidence shows that these latter are not readily degradable in marine sediments and can alter benthic assemblages. The potential impact of biodegradable bags on seagrasses inhabiting sandy bottoms, which are the most widespread and productive ecosystems of the coastal zones, has been ignored. Mesocosm experiments were conducted to assess the effect of a commercialized biodegradable bag on a common seagrass species of the Mediterranean, Cymodocea nodosa, both at the level of individual plant (clonal growth) and of plant community (plant-plant relationships), under three culture regimes (plant alone, in combination with a neighbour of the same species or of the co-existing seagrass Zostera noltei) simulating different natural conditions (bare substrate, monospecific meadows or mixed meadows). The bag behaviour in marine sediment and sediment physical/chemical variables were also examined. After six months of sediment exposure, the bag retained considerable mass (85% initial weight) and reduced sediment pore-water oxygen concentration and pH. In the presence of bag, C. nodosa root spread and vegetative recruitment increased compared to controls, both intra- and interspecific interactions shifted from neutral to competitive, and the growth form changed from guerrilla (loosely arranged group of widely spaced ramets) to phalanx form (compact structure of closed spaced ramets) but only with Z. noltei. These findings suggest that biodegradable bags altering sediment geochemistry could promote the spatial segregation of seagrass clones and influence species coexistence.

Non-biodegradable and compostable plastic bags on beaches negatively affect the development of coastal dune vegetation by drastically reducing seedling emergence and growth.

The presence of plastic bags on coastal dunes worldwide is well documented. Plastic bags contain additives that during rainfall events can leach out from bags into sand dune and be absorbed by seeds and roots of plants. Dune plants play a fundamental role in dune system formation, yet the possible impact of bag leaching on their establishment and development has been neglected. We assessed in laboratory whether (i) not biodegradable bags (high-density polyethylene, PE) and new generation of compostable bags (Mater-bi®, MB) would influence via leaching water chemical/physical properties and (ii) leachates would affect germination and seedling growth using Thinopyrum junceum and Glaucium flavum as models. Leachates were obtained from different amounts of not-exposed and bags exposed to beach or marine conditions simulating various pollution degrees (none, low, intermediate and high pollution). All water variables were affected by leaching. The magnitude of these alterations depended on bag type and environmental exposure. Seeds of T. junceum treated with the high concentration of marine-exposed MB bag leachate germinated later than controls while those of G. flavum treated with the remaining leachates germinated earlier. For both species, leachates from the low concentration of PE and MB marine-exposed bags increased seed germinability. A short radicle was observed in T. junceum seedlings treated with not-exposed MB bag leachates. Glaucium flavum seedlings treated with beach- and marine-exposed PE bags and not-exposed MB bags leachates showed a greater below-aboveground length ratio and those grown with the low concentration of not-exposed PE bag leachate had a longer hypocotyl compared to controls. Leachates from the high concentration of PE and MB bag caused seedling anomalies in both species. These findings indicate that not biodegradable and compostable bags may interact with abiotic/biotic factors and affect via leaching germination phenology, seedling establishment and plant interactions with consequences on dune community structure.

Propagule pressure is considered a major driver of plant invasion success. Great propagule pressure would enable invasive species to colonize new areas overcoming the resistance of native species. Many highly invasive aquatic macrophytes regenerate from vegetative propagules, but few studies have experimentally investigated the importance of propagule pressure and biotic resistance, and their interaction, in determining invasion success. By manipulating both recipient habitat and the input of vegetative propagules of the invasive seaweed Caulerpa cylindracea in mesocosm, we examined whether higher propagule pressure would overcome the resistance of a native congeneric (Caulerpa prolifera) and influence its performance. With the native, C. cylindracea population frond number decreased irrespectively of pressure level. High propagule pressure did not increase stolon length and single plant size decreased due to the effects of intra- and interspecific competition. Native biomass decreased with increasing C. cylindracea propagule pressure. These results indicate that higher propagule pressure may fail in enhancing C. cylindracea invasion success in habitats colonized by the native species, and they suggest that biotic resistance and propagule pressure co-regulate the invasion process. These findings emphasize the need to preserve/restore native seaweed populations and may help to design effective management actions to prevent further C. cylindracea spread.

Most researches on the plastisphere in coastal environments deal with plastics floating in seawater. Comparatively smaller attention has been devoted to the plastisphere of plastics buried in marine sediments, and very little is known on that of plastics on coastal sand dunes. Yet, limited information is available on the impact of plastics, especially biodegradable plastics, on microbial organisms in their surroundings. Nevertheless, a large amount of plastics sink on the seabed or is deposited on beach-dune systems. We investigated the succession of plastisphere microbial community on two biodegradable composites based on poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and seagrass fibres (PHBV/PO), buried in seabed and dune sediments over a 27 months period in mesocosm. PHBV is regarded as a valuable alternative to conventional plastics and PHBV/PO has recently been designed for applications in coastal habitat restoration. We also examined the degradation rate and impact of these plastics on the microbial communities of surrounding sediments. Microbial communities of the surface of PHBV and PHBV/PO in seabed and dune sand differ from those of surrounding sediments, displaying a lower richness. Plastics colonization occurs largely from bacteria present in surrounding sediments, although the contribution from the water column bacterial pool could be not negligible for plastics in the seabed. No significant differences were detected between the communities of the two plastics and no significant impact of plastics on microbial community of the surrounding sediments was detected. The exceptional long duration of this study allowed to gain information on the succession of a plastisphere community over a previously unexplored time scale. Succession appears highly dynamic in dune sand even after two years, while the community structure in seabed seems to reach stability after one year. These findings highlight the importance of performing long-term studies when trying to characterize composition and dynamics of plastisphere bacterial communities.

Background: The aim of this study was to provide an Italian version of the Haemophilia Activities List (HAL) and check its reliability in Italian medical centers. Methods: The Italian version of this assessment was administered to 80 patients (aged 18-65 years) affected by haemophilia A and B (moderate or severe). The validation was accomplished by comparing it to the revised and expanded Arthritis Impact Measurement Scales (AIMS2). Results: The internal consistency of the Italian version of the HAL had statistically high results: Cronbach's α 0.957-0.579. The highest internal consistency was measured in the domains ‘leg functionality' and in the overall points of the HAL questionnaire. The correlation between the AIMS2, which has been translated into Italian, and the version of the HAL questionnaire that we proposed, yielded good results for the following correlations: AIMS2 all and HAL overall (r = 0.64), AIMS2 physical function and HAL overall (r = 0.66), AIMS2 pain and HAL overall (r = 0.66). Conclusion: The Italian version of the HAL questionnaire presents both internal coherence and convergent validity. It can be used in addition to other functional tests to measure outcomes in moderate and severe haemophiliac diseases or to determine the quality of life as observed in the everyday life of patients.

Traditional revegetation techniques employed to restore seagrass meadows and coastal dunes have recently been criticized for their impact on donor populations as well as for the installation of plant anchoring structures made of non-biodegradable or not natural materials in recipient habitats. To improve the ecological sustainability of restoration practices, a novel plantable biodegradable container made of beach-cast seagrass wrack and a bio-based polymer was produced. The long-term performance of two seagrasses, Cymodocea nodosa and Zostera noltei, and two dune plants, Euphorbia paralias and Thinopyrum junceum, grown in nurseries from seeds using the bio-container or a non-biodegradable container of equal size/form made of a conventional plastic (control) was also examined. In addition, the development of bio-container-raised C. nodosa plants in the field was compared to that of plants removed from control containers at the installation and anchored with a traditional system. The bio-container degraded slowly in seawater and in sand and lost its functionality after about three years. In nurseries, all the tested species grown in bio-containers performed as well as, or better than, those raised in non-biodegradable ones. Six months after transplanting into the field, 80% of the C. nodosa nursery-raised plants installed with their bio-container have colonized the surrounding substrate while most of those planted with the traditional system was lost. These results indicate that the new bio-container may support plant growth, and it may also provide protection and anchorage to plants in the field. The use of this bio-container in combination with nursery techniques could improve the environmental sustainability of coastal restoration interventions by providing large plant stocks from seed, thus reducing the impact of collection on donor populations. This approach would also limit the introduction of extraneous materials in recipient habitats and offer an opportunity for valorizing seagrass beach-cast material.

In order to produce sustainable, bio-based and highly biodegradable materials, composites based on poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and fibers of Posidonia oceanica (PO), a dominant Mediterranean seagrass, were produced by simple melt mixing and characterized in terms of thermal stability, morphology and rheological/mechanical properties. In view of their potential application in marine environments, degradation of the developed composites was evaluated under simulated and real marine environmental conditions for 1 year. Using 10 wt % of acetyl tributyl citrate (ATBC) as a plasticizer, smooth processing was achieved for up to 30 wt % of PO fibers, despite the reduction of the melt fluidity observed with increasing fiber loading. The tensile modulus slightly increased (from 2 to 2.4 GPa) while the tensile strength and the elongation decreased (from 23.6 to 21.5 MPa and from 3.2 to 1.9%, respectively) by increasing the PO fiber content from 0 to 30 wt %. Interestingly, the impact resistance of the composites increased with the increasing of the PO content: the Charpy’s impact energy increased from 3.6 (without fiber) to 4.4 kJ/m2 for the composite with 30 wt %. The results of the aerobic biodegradation under simulated marine conditions showed that the presence of PO fibers favored the physical disintegration of the composite increasing the biodegradation rate of the polymeric matrix: after 216 days, the composite with 20 wt % PO fibers showed a biodegradability of about 30% compared to 20% of the composite without fibers. Under real marine conditions, the specimens containing PO fibers showed higher weight losses and deterioration of tensile properties compared to those without fibers. Presumably, biodegradation occurred after colonization of the specimen, and the specimens with 20 wt % PO fibers showed well-developed biofilm consisting of bacteria and fungi on the surface after only 3 months of incubation in marine sediments, unlike the no-fiber specimens. Consequently, the persistence of an adequate mechanical performance for a relatively long period (1 year), due to a moderate rate of biodegradation in the marine environment, make the developed PHBV/PO composites particularly suitable for the production of relatively low-cost and biodegradable items which are usable in the sea and/or sand dunes, increasing the market opportunities for biopolymers such as PHBV and, at the same time, finding an eco-sustainable valorization for the PO fibrous residues accumulated in large quantities on Mediterranean beaches, which represents a problem for coastal municipalities.

Root morphology and root system architecture of young Posidonia oceanica plants established on two contrasting substrate types, sand and rock, were examined to provide insights into the strategy of adaptation of seagrasses to their environment. After germination, seedlings were planted on sandy patches and on rock within the same area, and survived plants were collected five years later for measurements of the size of the entire root complex and analysis of individual morphological and architectural root traits. Collected plants exhibited up to nine highly intermingled root systems and approx. 2.5 m of total root length. Maximum horizontal extension, total biomass and total length of roots were not significantly affected by substrate. However, on sand roots grew vertically reaching up to 13 cm, while on rock they extended more horizontally and did not penetrate deeper than 5–7 cm leading to the formation of a shallow, densely packed root complex. On rock, the number and the length of second-order laterals on an individual root system were reduced and the topological index higher than on sand (0.8 vs. 0.7) reflecting a more simple (herringbone) branching pattern. Again, root diameter was greater than on sand. The results suggest that P. oceanica can adjust root traits early during plant development according to substrate typology to maximize anchorage and substrate exploration efficiency. This plasticity enables the species to establish and persist also on rocky bottoms which generally prevent establishment of the majority of seagrasses.

The effects of climate change are likely to be dependent on local settings. Nonetheless, the compounded effects of global and regional stressors remain poorly understood. Here, we used CO2 vents to assess how the effects of ocean acidification on the seagrass, Posidonia oceanica, and the associated epiphytic community can be modified by enhanced nutrient loading. P. oceanica at ambient and low pH sites was exposed to three nutrient levels for 16 months. The response of P. oceanica to experimental conditions was assessed by combining analyses of gene expression, plant growth, photosynthetic pigments and epiphyte loading. At low pH, nutrient addition fostered plant growth and the synthesis of photosynthetic pigments. Overexpression of nitrogen transporter genes following nutrient additions at low pH suggests enhanced nutrient uptake by the plant. In addition, enhanced nutrient levels reduced the expression of selected antioxidant genes in plants exposed to low pH and increased epiphyte cover at both ambient and low pH. Our results show that the effects of ocean acidification on P. oceanica depend upon local nutrient concentration. More generally, our findings suggest that taking into account local environmental settings will be crucial to advance our understanding of the effects of global stressors on marine systems.

Seagrasses are declining globally, and deeper understanding is needed on the recruitment potential and distribution of new populations for many threatened species to support conservation planning in the face of climate change. Recruitment of Posidonia oceanica, a threatened seagrass endemic to the Mediterranean, has long been considered rare due to infrequent flowering, but mounting evidence demonstrates that the species is responding to a changing climate through greater reproductive effort. Due to the fragmentary information on recruit occurrence and distribution, little is known about reproductive success in the species and its contribution to persistence. We assembled P. oceanica recruitment data from published and unpublished sources, including our own, to examine the frequency and extent of recruitment events (establishment of seedlings in a site), seedling growth potential and habitat characteristics at recruitment sites. Results show that at least one recruitment event has occurred about every three years, and eighteen localities were colonized at least one time since the first seedling record in 1986. Notably, consistently high seedling inputs were observed in four localities of the Western Mediterranean. Seedlings established mainly on unoccupied substrate areas along the coasts of islands, in sheltered sites and at shallower depths (

Bio-composites based on polyhydroxyalkanoates (PHAs) and fibres of Posidonia oceanica (PO) were investigated to assess their processability by extrusion, mechanical properties, and potential biodegradability in a natural marine environment. PHAs were successfully compounded with PO fibres up to 20 wt % while, at 30 wt % of fibres, the addition of 10 wt % of polyethylene glycol (PEG 400) was necessary to improve their processability. Thermal, rheological, mechanical, and morphological characterizations of the developed composites were conducted and the degradation of composite films in a natural marine habitat was evaluated in a mesocosm by weight loss measure during an incubation period of six months. The addition of PO fibres led to an increase in stiffness of the composites with tensile modulus values about 80% higher for composites with 30 wt % fibre (2.3 GPa) compared to unfilled material (1.24 GPa). Furthermore, the impact energy markedly increased with the addition of the PO fibres, from 1.63 (unfilled material) to 3.8 kJ/m2 for the composites with 30 wt % PO. The rate of degradation was markedly influenced by seawater temperature and significantly promoted by the presence of PO fibres leading to the total degradation of the film with 30 wt % PO in less than six months. The obtained results showed that the developed composites can be suitable to manufacture items usable in marine environments, for example, in natural engineering interventions, and represent an interesting valorisation of the PO fibrous wastes accumulated in large amounts on coastal beaches.

Este artículo contiene 16 páginas, 5 figurtas, 6 tablas, 3 apéndices con 3 tablas., The Biocoenosis of Well Sorted Fine Sands (WSFS) (SFBC, Sables Fins Bien Calibr es in French) is a Mediterranean community very well delimited by bathymetry (2e25 m) and sedimentology (>90% of fine sand) occurring in zones with relatively strong hydrodynamics. In this study focused on sites located along the Algerian, French, Italian and Spanish coasts of the Western Basin of the Mediterranean Sea (WBMS) we aim to compare the structure, ecological status and diversity of the macrofauna of the WSFS and examine the effects of recent human pressures on the state of this shallow macrobenthic community. We assess the ecological status and functioning of these WSFS using three categories of benthic indices: a) five indices based on classification of species into ecological groups, AMBI, BO2A, BPOFA, IQ and IP, b) the ITI index based on classification of species in trophic groups, and c) the Shannon H’ index, and the Biological Traits Analysis (BTA), which is an alternative method to relative taxon composition analysis and integrative indices. Cluster analyses show that each zone show a particular taxonomic richness and dominant species. The seven benthic indices reveal that the macrobenthos of the WSFS of the four coastal zones show good or high Quality Status, except for one location on the Algerian coast (the Djendjen site) in 1997. BTA highlights the presence of three groups of species: 1) typical characteristic species; 2) indicator species of enrichment of fine particles and organic matter, and 3) coarse sand species which are accessorily found on fine sand. Finally, the WSFS which are naturally subject to regular natural physical perturbations show a high resilience after human pressures but are very sensitive to changes in the input of organic matter., The authors thank the MEDISCAN programme for the Camargue Data, the Italian Research was funded by University of Pisa (Lardicci 308/2004) with the technical help of the Environmental Protection Regional Agency of Tuscany (ARPA Toscana).

Sediment fertilization is recommended for improving seagrass restoration efforts, but few studies have evaluated the efficacy of such practice. Increasing storm frequency due to global change could lead to greater sediment mobilization. Understanding how this alteration will interact with fertilization to affect transplants is essential for future restoration planning. We examined the individual and combined effects of nutrients (ambient vs. repeated addition) and burial (control vs. increased frequency and intensity) on the performance and biomass partitioning of transplants of the seagrass Cymodocea nodosa at two sites within a north-western Mediterranean meadow. Fertilization stimulated the production of shoots, total biomass, and branching. Burial increased leaf sheath length in one site while reduced shoot number, leaf number, leaf sheath length, total biomass, net shoot gain, and root-to-shoot ratio in the other site. Regardless of the site, fertilization and burial interaction reduced the length of vertical internodes and horizontal rhizomes, and the net shoot gain. Our research demonstrates that sediment fertilization ensures rapid colonization of restoration sites, providing C. nodosa plants up to eight times larger than controls in one growing season. However, it also indicates that interaction of increased burial and nutrients reduced the gain in terms of vegetative expansion and depressed vertical growth, making plants more vulnerable to subsequent disturbances. Therefore, seagrass restoration practitioners should account for changes in sediment elevation at transplanting sites when planning restoration programs and carefully evaluate the opportunity of applying fertilizers in sites subjected to greater sediment accumulation to avoid failure.

M.M. van Katwijk et al., In coastal and estuarine systems, foundation species like seagrasses, mangroves, saltmarshes or corals provide important ecosystem services. Seagrasses are globally declining and their reintroduction has been shown to restore ecosystem functions. However, seagrass restoration is often challenging, given the dynamic and stressful environment that seagrasses often grow in. From our world-wide meta-analysis of seagrass restoration trials (1786 trials), we describe general features and best practice for seagrass restoration. We confirm that removal of threats is important prior to replanting. Reduced water quality (mainly eutrophication), and construction activities led to poorer restoration success than, for instance, dredging, local direct impact and natural causes. Proximity to and recovery of donor beds were positively correlated with trial performance. Planting techniques can influence restoration success. The meta-analysis shows that both trial survival and seagrass population growth rate in trials that survived are positively affected by the number of plants or seeds initially transplanted. This relationship between restoration scale and restoration success was not related to trial characteristics of the initial restoration. The majority of the seagrass restoration trials have been very small, which may explain the low overall trial survival rate (i.e. estimated 37%). Successful regrowth of the foundation seagrass species appears to require crossing a minimum threshold of reintroduced individuals. Our study provides the first global field evidence for the requirement of a critical mass for recovery, which may also hold for other foundation species showing strong positive feedback to a dynamic environment. Synthesis and applications. For effective restoration of seagrass foundation species in its typically dynamic, stressful environment, introduction of large numbers is seen to be beneficial and probably serves two purposes. First, a large-scale planting increases trial survival - large numbers ensure the spread of risks, which is needed to overcome high natural variability. Secondly, a large-scale trial increases population growth rate by enhancing self-sustaining feedback, which is generally found in foundation species in stressful environments such as seagrass beds. Thus, by careful site selection and applying appropriate techniques, spreading of risks and enhancing self-sustaining feedback in concert increase success of seagrass restoration. For effective restoration of seagrass foundation species in its typically dynamic, stressful environment, introduction of large numbers is seen to be beneficial and probably serves two purposes. First, a large-scale planting increases trial survival - large numbers ensure the spread of risks, which is needed to overcome high natural variability. Secondly, a large-scale trial increases population growth rate by enhancing self-sustaining feedback, which is generally found in foundation species in stressful environments such as seagrass beds. Thus, by careful site selection and applying appropriate techniques, spreading of risks and enhancing self-sustaining feedback in concert increase success of seagrass restoration. Journal of Applied Ecology, A.T. was funded by Greater Caribbean Energy and Environment Foundation grants. N.M. was supported by a Gledden Fellowship from the Institute of Advanced Studies of the University of Western Australia. N.M. C.M.D and A.C. were supported by Biomares contract number LIFE06 NAT/PT/000192. N.M. and C.M.D. were supported by Opera (FP7, contract number 308393). C.P. and the Cornell Cooperative Extension Marine Program are funded in part by County Executive Steve Bellone and the Suffolk County Legislature, Hauppauge, New York. E.B. and C.L. were funded by University of Pisa (Lardicci 308/ex60%2010). M.L.C and G.A.K were supported by ARC Linkage Grants (LP130100155, LP0454138)

Exogenous application of plant growth regulators (PGRs) may be an effective technique for increasing the rooting ability and the growth of vegetative fragments (cuttings) of plants used in dune restoration programs. Various concentrations (0, 50 and 100 mg l −1 ) of two auxins, alpha-naphtaleneacetic acid (NAA) and indole-3-butyric acid (IBA), and two cytokinins, 6-furfurylaminopurine (Kinetin) and 6-benzylaminopurine (BAP), were applied separately to cuttings of two widely used species for restoration, Ammophila arenaria and Sporobuls virginicus . Root development and production of new buds in cuttings were examined under laboratory conditions one month after application. Cuttings were also examined one year after transplanting into a sandy substratum under natural conditions, to test for possible long term effects of PGRs on plant establishment success and growth. The response of the two study species to PGRs differed substantially. In A. arenaria the auxin NAA at 100 mg l −1 reduced the time for root initiation and increased the rooting capacity of cuttings, while the cytokinin Kinetin at 50 mg l −1 facilitated root growth. No auxin had effect on rooting or growth of S. virginicus cuttings, but treatment with 100 mg l −1 Kinetin resulted in higher rooting success than the control. One year after planting, the cuttings of A. arenaria treated with 100 mg l −1 NAA showed a higher establishment success (90% vs. 55%) and produced more culms and longer roots than the control; those treated with cytokinins did not differ in the establishment success from the control, but had longer roots, more culms and rhizomes. On the other hand, the cuttings of S. virginicus treated with 100 mg l −1 Kinetin showed a higher establishment success (75% vs. 35%) and had more culms than the control. Therefore, in restoration activities that involved A. arenaria, a pre-treatment of cuttings with NAA would be beneficial, as it allows the production of a higher number of well-developed plants with high survival potential and greater area cover. In contrast, a pre-treatment of cuttings of S. virginicus with Kinetin would achieve more acceptable plant survival rates. This easy and low cost-effective technique may be extended to other dune plant species and applied on a large scale to improve the chance of dune restoration success.

[Purpose] The aim of the present study was to establish up-to-date data regarding the lung function of cystic fibrosis (CF) patients. [Subjects and Methods] Forty-eight patients of both genders, with a diagnosis of CF, were recruited. As a result our sample presented, according to the GOLD criteria, 23 patients with mild lung obstruction (FEV1%pred: 89.86), 16 patients with moderate lung obstruction (FEV1%pred: 56.1) and 9 patients with severe obstruction (FEV1%pred: 32.1). [Results] All patients presented normal total lung capacity followed by an important residual volume increase. [Conclusion] Our results were important to illustrate the CF patient’s lung functional status and to improve the health system strategy in treating such individuals.

The use of fragments detached naturally from Posidonia oceanica , a threatened seagrass protected by European legislation, as non-destructive alternative to removing cuttings from donor meadows for restoration efforts has been recently proposed. A 4-year study was performed to investigate whether storm-generated fragments deposited on beaches can be salvaged and used as transplant source. Specifically, we examined in outdoor tanks (i) how long fragments survive after detachment from parents, (ii) whether fragment survival and regeneration ability are influenced by morphological characteristics, i.e., initial fragment growth form and size, and (iii) whether a single fragment can be divided artificially to obtain more transplants. Finally, we monitored the first year survival of fragments lodged between rubbles of a submersed artificial reef 3 years after storage in tanks to test for their capacity to re-establish in the field. Four months after collection, fragment survival was high (80–96%). Half of the fragments regenerated and survived as long as 3 years in tanks. Initial growth form and size were not predictors of fragment survival or regeneration probability. Division of fragments allowed doubling transplant number. One year after transplanting, 50% of the fragments on the artificial reef were still present and ca. half of them produced new shoots. This indicates that fragments retain the capacity to re-establish once reintroduced into the field and thus could represent a potentially important transplant source. The use of this material could have major advantages over traditional restoration techniques, including large availability with zero impact on existing populations and low collection efforts.

In this study the occurrence of twin-like embryos and abnormal embryos in Posidonia oceanica , a rare phenomenon in seagrasses, was documented. The ability of additional embryos to develop seedlings was also demonstrated for the first time in seagrasses. Approximately 1750 fruits collected at three localities of the north-western Mediterranean on 2 years (1994 and 2004) were screened for abnormal embryo morphology. The frequency of embryo anomalies varied from 1.9 to 7.1% among localities, and no differences between years were detected within the same locality. Twin-like embryos were usually in contact and germinated to produce complete twin seedlings that could be separated. Abnormal embryos showed additional plumules but had a common hypocotyl and a single primary root; these embryos germinated and grew like “Siamese” seedlings.

A recruitment event of Posidonia oceanica was recorded in the Gulf of Ventilegne (Corsica, Western Mediterranean) in June 2005. One-year seedlings (alive and dead) were found at shallow depth (2–3 m) on a mix of rock and sand substrates. Pattern in the distribution of seedlings was examined over spatial scales ranging from centimetres to hundreds of metres. We also examined (a) whether establishment was preferentially associated with a specific substrate type; (b) the relation between proportional mortality and the density of established seedlings; and (c) the growth characteristics of seedlings. The average density of seedlings varied from 19 to 189 seedlings m−2, with a maximum of 480 seedlings m−2. While significant variation was detected at the scale of area (i.e., tens of metres), no difference was found in abundance of seedlings at the scale of sites (i.e., hundreds of metres). Most of the overall variability was, however, at on the smallest scale examined (i.e., among sampling units tens of centimetres apart). No differences were also observed in the establishment probability and seedling abundance between sand and rock substrates. The fine scale analysis of spatial patterns on rock and sand substrates showed a clumped distribution of seedlings. These results indicated that the spatial distribution of seedlings was not random, but organized at small spatial scales within the study sites. Hydrodynamics and micro-scale bottom features appeared to account for such patterning. Proportional seedling mortality was highly variable (ranging from 0.3 to 0.8), and tended to increase with increasing settlement density. A total of 128,000 seedlings were recoded over the 1600 m2 surface area investigated. Such event reflected conspicuous seed supply and particularly favourable local conditions. This finding contradicts the current statement of low recruitment ability of P. oceanica and indicates that the species can recruit both on consolidated and unconsolidated substrates, at least in the study area.

The intensive reconstructive sampling (1957-2004, 39 localities), a systematic direct observation (1992-2004, 1 locality) and particular direct observations (66 localities) of Posidonia oceanica meadows were analysed together with temporal series of flowering available in the literature (19 localities). This allowed the examination of temporal and spatial variability in annual flowering prevalence (FP, the fraction of meadows flowering in a given year) and of annual meadow flowering intensity (FI, number of inflorescences per shoot) for the period 1979-2004 across the Western Mediterranean, as well as spatial variability of flowering frequency (FF, the fraction of years that a given meadow has flowered) and shoot flowering probability (Pf, fraction of flowering stalks appeared per annual segment). Each year, on an average 17% of the investigated meadows flowered, ranging from 3 to 86% of meadows among the years. The highest annual FP and FI values were obtained in 2003 (FP = 0.86 and mean FI = 0.23 ± 0.03 inflorescences shoot-1). A secondary peak of FP and mean FI occurred 9 years earlier, in 1994 (FP = 0.44 and mean FI = 0.08 ± 0.02). Both peaks of flowering occurred after hot summers. Flowering synchrony in particular years across the Western Mediterranean and clines of increased meadow flowering frequency towards the North and East, suggests the existence of large-scale environmental mechanisms controlling the floral induction. On the other hand, meadow FF and Pf were highly heterogeneous among and within the meadows, indicating that local factors also may play a significant role in flowering induction. When flowering, the Western Mediterranean meadows showed an average 0.11 ± 0.02 inflorescences shoot-1, but FI greatly varied among and along the series (from 0.002 to 0.54 inflorescences shoot-1) and decreased significantly with depth but was independent of meadow shoot density and meadow latitude or longitude. The shoot flowering probability was quite low (0.007 ± 0.002 inflorescences shoot-1 year-1) and exponentially increased with shoot age. © Springer-Verlag 2005.

The seagrass Posidonia oceanica (L.) Delile can produce plagiotropic (horizontally oriented) and orthotropic (vertically oriented) axes. These axes differ in various morphological and growth characteristics and ecological functions. Only orthotropic shoots are likely to support flowering and seed production. To asses whether flowering may occur on plagiotropic shoots, we examined plagiotropic axes that grew at the edge of a meadow during the flowering of 2003. We identified and analysed 10 rhizome fragments that bore at least one reproductive plagiotropic shoot. We then investigated whether reproductive plagiotropic shoots differed from orthotropic ones in morphological characteristics and investment in flowering (i.e., number of hermaphroditic flowers per inflorescence). Multivariate analysis did not show any significant difference between plagiotropic and orthotropic shoots with regard to the characteristics examined and investment in flowering. Further studies are necessary to ascertain if the observed pattern of flowering is a common or an exceptional event in P. oceanica.

Urban and industrial wastes have been claimed to negatively affect Posidonia oceanica meadows, but few studies have addressed this issue by comparing disturbed locations with replicated reference locations. Here, we examined the general proposition that patterns of growth and morphology of P. oceanica exposed to urban and industrial effluents were different from those observed in reference meadows. Hypotheses were both on differences in mean values of response variables and on variation of these measures at a hierarchy of spatial scales (from centimetres to hundreds of metres). Results indicated a significant reduction in mean number of intermediate leaves at the outfall compared to reference locations, whereas the opposite pattern occurred for juvenile leaves. There were significant, though temporally variable differences in growth of rhizomes between disturbed and reference locations, with reduced growth at the outfall in 2 out of 3 years analysed. Measures of spatial variance in number of juvenile leaves and length of adult leaves at the scale of shoots were significantly larger at the outfall compared to reference locations. At the same scale, measures of spatial variance in length of juvenile and intermediate leaves were significantly lower at the putatively impacted location. Spatial variance in number of intermediate leaves was reduced at the outfall compared to reference locations at the scale of quadrats. Past values of spatial variance in number of leaves per shoot were lower at the outfall than at the two reference locations at the scale of shoots, whereas the opposite occurred at the scale of areas. None of the structural variables examined showed any difference between the putatively impacted location and the two reference locations, either in terms of mean response or as changes in spatial variance. These results indicated that integrating methods to examine present and past events of disturbance, including analyses to detect changes in spatial variance of response variables, may provide a powerful approach to the analysis of environmental impacts on P. oceanica.

There is a scarcity of data on the seedling establishment success of plants inhabiting the sand coasts of the Mediterranean, despite on increasing interest in the conservation and restoration of dune habitats. A field experiment, in conjunction with a series of laboratory germination tests, was conducted to (1) determine the recruitment capacity of Pancratium maritimum, a perennial dune plant which is declining along the north-western Italian coasts, by natural seed dispersal and (2) test the ability of seeds to tolerate different environmental stress. We planted seeds at different burial depths and found that seeds from greater depths (6, 10 and 15 cm) showed higher emergence (35.0–46.6%) than seeds from a shallower depth (2 cm; 11.6%). On average, less than 18% of seeds remained dormant at the end of the germination period. None of the seedlings that emerged in autumn survived over winter, and 88% of those that emerged in spring died by the start of the second reproductive season as a result of...

To identify the factors that might contribute to limiting the sexual reproduction of Posidonia oceanica we examined: (1) flowering and fruiting phenology; (2) variability in flowering frequency, seed production and reproductive success (i.e. the proportion of flowers setting mature fruits per unit area) over a 2-year period; and (3) losses of potential seed production to fruit abortion and/or pre-dispersal seed predation. The flowering frequency of P. oceanica varied among years, ranging between 7.9 and 19.8%. Despite the large number of flowers and ovules, few of these produced mature fruits owing to abortion. Moreover, about 84% of developing inflorescences were damaged by herbivores. Fruit production varied among years, ranging from 4.7 to 13.5 fruits per m2, but reproductive success remained constant (2.0–2.4%). Exclusion of herbivores showed that abortion alone was reponsible for the loss of about 87% of the reproductive potential. Since reduced fertilization did not completely explain the observed frequency of abortion, it was hypothesized that post-fertilization factors could affect seed formation. The higher reproductive success of protected plants (11.2%) compared to herbivore-exposed plants (3.1%) indicated that predation significantly reduced the number of seeds available for establishment. We concluded that pre-dispersal seed losses to abortion and predation may seriously reduce the reproductive success of the species.

In seagrasses, elongation of horizontal rhizomes, and hence clone expansion, is supported by physiological integration. However, horizontal rhizomes are subjected to breakage by physical disturbances and to burial, often simultaneously. Little is known about the combined impact of such factors on the growth of apical rhizome regions. We factorially manipulated connections on runners of Cymodocea nodosa and sediment height for 1 mo at 2 sites within a meadow to test the hypothesis that breakage of interconnections close (10 cm) to the rhizome apex causing premature loss of physiological integration would negatively affect the performance of runners and reduce their ability to tolerate burial (60% of shoot height). Shoot survival, length of vertical internodes and horizontal rhizome network, biomass production, partitioning, and costs were unaffected by the investigated factors. Rhizome breakage interacted with burial for some characteristics. Breakage reduced branch length and rhizome diameter only in unburied runners, while burial reduced branch length only in intact runners. Moreover, burial stimulated leaf sheath elongation and greater branching, irrespective of the status of connections. These findings indicate that breakage only delays lateral spread and increases the risk of fragmentation by subsequent disturbances, and, contrary to our hypothesis, has no substantial influence on the capacity of runners to withstand transitory and moderate burial. The findings also reveal that runners may escape from burial not only vertically, but also horizontally, changing growth pattern and direction from predominantly unidirectional to multidirectional. This additional strategy could allow buried runners to accelerate space occupation even if disconnected from clones.

A method for the isolation of protoplasts from the seagrasses Posidonia oceanica and Cymodocea nodosa is described. Isolation of protoplasts was achieved using a combination of cellulase Onozuka R-10, hemicellulase and pectinase. Purification was carried out by Ficoll gradient centrifugation. A yield of 1.87 (± 0.16 SD) × 10 6 protoplasts per gram of fresh tissue was obtained from mesophyll cells of P. oceanica. The viability of isolated protoplasts was 82.5% (± 10.6) as confirmed by fluorescein diacetate staining. A high percentage of protoplasts of P. oceanica (61.5 ± 14.8%) regenerated the cell wall within 7 days as confirmed by staining with calcofluor white, but only a few protoplasts were able to divide. Callus-like structures were noticed after 20–30 days in culture. A lower yield of protoplasts, 6.9 (± 3.8) × 10 5 protoplasts per gram of fresh tissue, was obtained from mesophyll of C. nodosa. Viability of these protoplasts was 67.9% (±12.6) after isolation. Some possible applications of the method are discussed. © 2001 Elsevier Science B.V. All rights reserved.

Two pilot transplant experiments were conducted in a damaged coastal area of the Ligurian Sea (Italy) to examine the feasibility of a large-scale restoration project. Two sites were selected in the area and two-month-old aquarium-grown Posidonia oceanica (L.) Delile seedlings were transplanted at each site on different substrata. Survival and development of seedlings were followed every month. The effects of substratum type (`matte' vs. pebbles) and herbivore exclosure on seedling success were tested approximately after three years. Survival and growth of natural P. oceanica seeds germinated in the study area were also investigated. Sites did not differ significantly in the survival and development of seedlings. The highest survival and development was observed for seedlings growing on `matte', with survival rates of about 70% for transplanted and 66% for natural seedlings. Growth characteristics of natural and transplanted seedling populations were similar. Herbivores did not affect the survival and development of transplanted seedlings. These results indicate that P. oceanica seedlings may be successfully used in the study area for restoration purposes. Furthermore, our data support the hypothesis that sexual recruitment may occur in P. oceanica but could be limited by postsettlement factors relating to substratum type, in the study area.

The effects of culture media and fruit storage on germination, development and survival of seedlings of Posidonia oceanica (L.) Delile (Posidoniaceae) were investigated under axenic conditions. Seeds germinated within 15–18 days both on solid media and in liquid over solid media as well as on filter-paper bridges over liquid media. The results showed a significant effect of the culture medium on germination and survival of seedlings among the solid media and the liquid over solid media tested. The highest mean germination (96.2%) was found in the solid artificial seawater medium (ASW). Only solid or liquid/solid ASW medium, however, allowed continuous growth of seedlings. Addition of nutrients and inclusion of the plant growth regulator 2,4-dichlorophenoxyacetic acid (2,4-D) inhibited development of seedlings. After 10 months, the highest mean survival (75%) was reached on solidified ASW following addition of liquid medium. No significant effect of the culture medium on seed germination and seedling survival was seen using the paper-filter bridge technique. With the latter method, germination and survival rates ranged from 54 to 85% and from 31 to 58%, respectively. All seedlings showed continuous growth. Addition of nutrients, however, reduced the production of adventitious roots. The results also showed significant effect of fruit storage treatment on the fruit surface-sterilization success, but not on seed germination or seedling survival. In vitro raised seedlings of P. oceanica appear to be useful for conservation and propagation of this species.

SummaryThe genome of Posidonia oceanica (Potamogetonaceae) was studied at cytophotometric and biochemical level. P. oceanica is an important species for food web in the Mediterranean basin where it occupies very large prairies. It is also an important biological indicator of mercury contamination. Cytophotometric analyses showed that this species has a basic (4C) genome size of 11.16 pg DNA. Biochemical analyses were made to establish genome composition in terms of highly and medium repeated and unique sequences. Melting profiles and CsCl density gradient of DNA extracted from leaves indicated the presence of a large A-T rich satellite DNA.