Due to multiple impacts, Cystoseira forests are experiencing a significant decline, which is affecting the ecosystem services they provide. Despite conservation efforts, there is an urgent need to develop best practices and large-scale restoration strategies. To implement restoration actions, we developed an ex situ protocol for the cultivation of Cystoseira. amentacea var. stricta, aimed at reducing the time needed for laboratory culture, thus avoiding prolonged maintenance and minimizing costs. Specifically, we tested the effects of temperature, light and substratum on settlement and growth of early life stages using a factorial experiment. Temperature (20 and 24°C) and photoperiod (15L:9D) were selected to reflect the conditions experienced in the field during the reproductive period. Two light intensities (125 and 250 μmol photons m−2s−1) were selected to mimic the condition experienced in the absence of canopy (i.e. barren—higher light intensity) or in the understory (lower light intensity) during gamete release. The tested substrata were flat polished pebbles and rough clay tiles. The release of gametes and the successive survival and development of embryo and germlings were followed for two weeks. Regardless of the culture conditions, rougher tiles showed higher zygote settlement, but the substrata did not affect the successive development. Zygote mortality after one week averaged 50% and at the end of the second week, embryonic survival was higher under lower light and temperature conditions, which also determined the growth of larger embryos.

In the Mediterranean Sea, brown algae belonging to the Cystoseira genus play a valuable role as foundation species. Due to evidences of regression/loss of the habitats of these species caused by the interplay of human and climatic disturbances, active restoration measures have been encouraged by EU regulations. In particular, nondestructive restoration techniques, which avoid the depletion of threatened species in donor populations, are strongly recommended. In the framework of the EU project ROCPOP-Life, the first ex situ outplanting experience of Cystoseira amentacea var. stricta has been implemented in the Cinque Terre Marine Protected Area (northwestern Mediterranean). A total of 400 clay tiles, hosting approximately three mm-long germlings of C. amentacea, were fixed to the rocky shore with screws: the tiles were monitored for the next 2 months by photographic sampling, and survival (presence/absence of juveniles on the tiles), cover and growth were assessed. Additional sampling was performed 6 months after tile deployment, after which an unprecedented storm surge severely affected the restoration performance. After 2 months, over 40% of the tiles were covered with Cystoseira juveniles, which reached approximately eight mm in total length. The tiles that survived the storm hosted three to six cm-long juveniles. The high cover (≥25%), assuring moisture and shading, and the appropriate size of the juveniles, to avert micro-grazing, at time of deployment were key to the survival and growth of the outplanted juveniles, increasing the potential for restoration success. Our findings show that outplanting of midlittoral canopy-forming species is a feasible approach for restoration efforts, with particular attention given to the early phases: (i) laboratory culture, (ii) transport, and (iii) juvenile densities. These results are strongly encouraging for the implementation of restoration actions for C. amentacea on a large scale, in light of EU guidelines.

1. The worldwide decline of marine forests, due to human impacts and climate change, emphasizes the need to develop and implement effective and sustainable solutions to restore these endangered habitats and to re-establish the services they provide.2. In this study, the ex situ restoration of Gongolaria barbata (=Treptacantha barbata), a Mediterranean subtidal habitat-forming species of brown seaweed, was for the first time implemented in a marine protected area in the Adriatic Sea. Two restoration efforts were performed in 2019. The first was started in winter, after a marine heatwave that triggered early fertility, and the second in spring, when the species usually reproduces.3. This study aimed to evaluate: (i) the disruptive effects of a thermal anomaly on the reproductive biology and performance in culture of G. barbata; and (ii) the impact of the grazing pressure on juveniles after the outplanting. 4. The first cultivation was more productive than the second, in terms of zygote release and germling growth. To mitigate the low efficiency of the second culture and to avoid prolonged highly demanding maintenance in the mesocosms, the cultivation period was extended outdoors using a structure suspended in the water column.5. The modular frames conceived for outplanting G. barbata proved to be effective because of their easy operability and low cost. Controlling for herbivorous fish had significant positive effects on both juvenile survival and growth.6. The outcomes highlighted that an unpredictable climatic event and fish grazing were major threats that impaired the restoration process of G. barbata. These stressors should be considered when developing plans to implement effective large-scale restoration of canopy-forming macroalgae.

Inflammation and oxidative stress are part of the complex biological responses of body tissues to harmful stimuli. In recent years, due to the increased understanding that oxidative stress is implicated in several diseases, pharmaceutical industries have invested in the research and development of new antioxidant compounds, especially from marine environment sources. Marine seaweeds have shown the presence of many bioactive secondary metabolites, with great potentialities from both the nutraceutical and the biomedical point of view. In this study, 50%-ethanolic and DMSO extracts from the species C. amentacea var. stricta were obtained for the first time from seaweeds collected in the Ligurian Sea (north-western Mediterranean). The bioactive properties of these extracts were then investigated, in terms of quantification of specific antioxidant activities by relevant ROS scavenging spectrophotometric tests, and of anti-inflammatory properties in LPS-stimulated macrophages by evaluation of inhibition of inflammatory cytokines and mediators. The data obtained in this study demonstrate a strong anti-inflammatory effect of both C. amentacea extracts (DMSO and ethanolic). The extracts showed a very low grade of toxicity on RAW 264.7 macrophages and L929 fibroblasts and a plethora of antioxidant and anti-inflammatory effects that were for the first time thoroughly investigated. The two extracts were able to scavenge OH and NO radicals (OH EC50 between 392 and 454 μg/mL; NO EC50 between 546 and 1293 μg/mL), to partially rescue H2O2-induced RAW 264.7 macrophages cell death, to abate intracellular ROS production in H2O2-stimulated macrophages and fibroblasts and to strongly inhibit LPS-induced inflammatory mediators, such as NO production and IL-1α, IL-6, cyclooxygenase-2 and inducible NO synthase gene expression in RAW 264.7 macrophages. These results pave the way, for the future use of C. amentacea metabolites, as an example, as antioxidant food additives in antiaging formulations as well as in cosmetic lenitive lotions for inflamed and/or damaged skin.

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