Comparative evaluation of multiple protein extraction procedures from three species of the genus Caulerpa

AbstractThe aim of this study was to define the simplest and least expensive protocol for total protein extraction for three different macroalgae of the genus Caulerpa (the invasive C. taxifolia and C. cylindracea and the autochthonous C. prolifera). Five multi-step protein extraction procedures, set up for other macroalgal species, were tested. For each of them, different pre-treatment and extraction conditions were simultaneously examined, according to a factorial design, considering the starting material, the solvent-to-biomass ratio, and the incubation temperature. Protein yield in the obtained extracts was estimated with the Bradford method. Further, polyacrylamide gel electrophoresis (SDS-PAGE) was used to resolve proteins, assessing their quality and integrity. Significant differences in protein yield were observed among the extraction protocols and the conditions tested, also in relation to the considered species. Profiles having an acceptable quality were obtained for C. prolifera and C. cylindracea, and from the obtained results, the best method to obtain high yield and quality protein extracts for the two above-mentioned species appears to require the use of a primary TCA/acetone extraction buffer followed by a lysis buffer with NaCl, KCl, urea, Triton, SDS and a protease inhibitor. The best results, in particular, were obtained starting from fresh pulped material with a buffer-to-biomass ratio of 10:1 and an incubation temperature of 4°C. For C. taxifolia, instead, none of the tested protocols produced satisfactory results and further studies will be required

Untangling natural variability of macrofaunal populations from protection effects

Marine protected areas are essential for conservation purposes. Remarkably, few studies have assessed the responses of small macrofaunal species to different protection levels in the Mediterranean Sea. Using a hierarchical sampling design spanning four orders of magnitude (1, 10, 100 and 1000 m), we investigated whether a marine protected area had an effect on crustacean (amphipod) and mollusc (mainly gastropod) assemblages associated with Posidonia oceanica seagrass meadows. Based on this investigation, we report spatial and temporal variability patterns of these assemblages in four different protection levels. We also discuss potential confounding effects on these areas, such as different habitat features. The structure of amphipod and gastropod assemblages based on density data was patchy at small and large spatial scales, and differed markedly among protection levels. Multiscale analyses showed that lower densities and/or biomasses of several taxa occurred within fully protected areas when compared to partially protected areas. Moreover, the seagrass meadows accounted for a low proportion of the total variability of the studied macrofaunal assemblages. We suggest therefore that the observed patchiness is likely to occur for multiple and interrelated reasons, ranging from the ecological and behavioural traits (e.g., dispersion and mobility) of macrofaunal species to protection-dependent factors such as fish predation. Multiscale spatial and temporal monitoring of macrofaunal assemblages in a long-term perspective, as well as experimental manipulations, are needed to better understand the protection effects

Consumer depletion alters seagrass resistance to an invasive macroalga

Few field studies have investigated how changes at one trophic level can affect the invasibility of other trophic levels. We examined the hypothesis that the spread of an introduced alga in disturbed seagrass beds with degraded canopies depends on the depletion of large consumers. We mimicked the degradation of seagrass canopies by clipping shoot density and reducing leaf length, simulating natural and anthropogenic stressors such as fish overgrazing and water quality. Caulerpa racemosa was transplanted into each plot and large consumers were excluded from half of them using cages. Potential cage artifacts were assessed by measuring irradiance, scouring by leaf movement, water flow, and sedimentation. Algal invasion of the seagrass bed differed based on the size of consumers. The alga had higher cover and size under the cages, where the seagrass was characterized by reduced shoot density and canopy height. Furthermore, canopy height had a significant effect depending on canopy density. The alteration of seagrass canopies increased the spread of C. racemosa only when large consumers were absent. Our results suggest that protecting declining habitats and/or restoring fish populations will limit the expansion of C. racemosa. Because MPAs also enhance the abundance and size of fish consuming seagrass they can indirectly promote algal invasion. The effects of MPAs on invasive species are context dependent and require balancing opposing forces, such as the conservation of seagrass canopy structure and the protection of fish grazing the seagrass

Using complementary visual approaches to investigate residency, site fidelity and movement patterns of the dusky grouper (Epinephelus marginatus) in a Mediterranean marine protected area

AbstractAssessing individuals' abundance, residency (presence at a site within a certain period) and site fidelity (tendency to return to the same site in subsequent seasons or years) is crucial for evaluating and improving the effectiveness of spatial conservation/management measures regarding ecologically and socio-economically valuable species. Using underwater visual census (UVC) and photo-identification (photo-ID) techniques, we estimated the abundance, residency and site fidelity of the dusky grouper, Epinephelus marginatus, at two protected sites within the Tavolara-Punta Coda Cavallo Marine Protected Area (Sardinia, Italy) in the summers of 2017–2018. The scope and spatio-temporal resolution of the study was extended by involving volunteer recreational divers in the photo collection. Grouper mean densities varied significantly across sampling dates, with a significant variability between the 2 years and the two investigated sites. At least 94 grouper visited the study sites in the summers of 2017–2018 based on the analysis of 968 high-quality photos using a semi-automated software to photo-identify individuals. Overall, the most frequently sighted grouper was recorded on 32 different days and 21 individuals (22%) identified in 2017 were re-sighted in 2018. The participation of volunteer recreational divers helped detect the inter-site (3.5–4 km apart) movements of a female and a male, supporting previous findings regarding the occurrence of reproduction-related movements. This study provides novel insights into the residency and site-fidelity patterns of the dusky grouper, and its small-scale movements probably related to reproduction. Specifically, we provide indications that effective protection from fishing should encompass the entire area used by grouper for reproductive movements

Large-scale assessment of Mediterranean marine protected areas effects on fish assemblages

Marine protected areas (MPAs) were acknowledged globally as effective tools to mitigate the threats to oceans caused by fishing. Several studies assessed the effectiveness of individual MPAs in protecting fish assemblages, but regional assessments of multiple MPAs are scarce. Moreover, empirical evidence on the role of MPAs in contrasting the propagation of non-indigenous-species (NIS) and thermophilic species (ThS) is missing. We simultaneously investigated here the role of MPAs in reversing the effects of overfishing and in limiting the spread of NIS and ThS. The Mediterranean Sea was selected as study area as it is a region where 1) MPAs are numerous, 2) fishing has affected species and ecosystems, and 3) the arrival of NIS and the northward expansion of ThS took place. Fish surveys were done in well-enforced no-take MPAs (HP), partially-protected MPAs (IP) and fished areas (F) at 30 locations across the Mediterranean. Significantly higher fish biomass was found in HP compared to IP MPAs and F. Along a recovery trajectory from F to HP MPAs, IP were similar to F, showing that just well enforced MPAs triggers an effective recovery. Within HP MPAs, trophic structure of fish assemblages resembled a top-heavy biomass pyramid. Although the functional structure of fish assemblages was consistent among HP MPAs, species driving the recovery in HP MPAs differed among locations: this suggests that the recovery trajectories in HP MPAs are likely to be functionally similar (i.e., represented by predictable changes in trophic groups, especially fish predators), but the specific composition of the resulting assemblages may depend on local conditions. Our study did not show any effect of MPAs on NIS and ThS. These results may help provide more robust expectations, at proper regional scale, about the effects of new MPAs that may be established in the Mediterranean Sea and other ecoregions worldwide

Large-Scale Assessment of Mediterranean Marine Protected Areas Effects on Fish Assemblages

Marine protected areas (MPAs) were acknowledged globally as effective tools to mitigate the threats to oceans caused by fishing. Several studies assessed the effectiveness of individual MPAs in protecting fish assemblages, but regional assessments of multiple MPAs are scarce. Moreover, empirical evidence on the role of MPAs in contrasting the propagation of non-indigenous-species (NIS) and thermophilic species (ThS) is missing. We simultaneously investigated here the role of MPAs in reversing the effects of overfishing and in limiting the spread of NIS and ThS. The Mediterranean Sea was selected as study area as it is a region where 1) MPAs are numerous, 2) fishing has affected species and ecosystems, and 3) the arrival of NIS and the northward expansion of ThS took place. Fish surveys were done in well-enforced no-take MPAs (HP), partially-protected MPAs (IP) and fished areas (F) at 30 locations across the Mediterranean. Significantly higher fish biomass was found in HP compared to IP MPAs and F. Along a recovery trajectory from F to HP MPAs, IP were similar to F, showing that just well enforced MPAs triggers an effective recovery. Within HP MPAs, trophic structure of fish assemblages resembled a top-heavy biomass pyramid. Although the functional structure of fish assemblages was consistent among HP MPAs, species driving the recovery in HP MPAs differed among locations: this suggests that the recovery trajectories in HP MPAs are likely to be functionally similar (i.e., represented by predictable changes in trophic groups, especially fish predators), but the specific composition of the resulting assemblages may depend on local conditions. Our study did not show any effect of MPAs on NIS and ThS. These results may help provide more robust expectations, at proper regional scale, about the effects of new MPAs that may be established in the Mediterranean Sea and other ecoregions worldwide

The interactive effect of herbivory, nutrient enrichment and mucilage on shallow rocky macroalgal communities

This paper focuses on the interactive short and long-term effect of three different stressors on a macroalgal assemblage. Three stressors are considered: herbivory, nutrients and mucilage. The experiment was conducted in Tavolara Punta Coda Cavallo Marine Protected Area (Mediterranean Sea) during a bloom of the benthic mucilage-producing microalga Chrysophaeum taylorii (Pelagophyceae); this microalga is recently spreading in the Mediterranean Sea. On a rocky substratum, 36 plots 20 × 20 cm in size were prepared. Factorial combinations of three experimental treatments were applied in triplicate, including three grazing levels crossed with two nutrient enrichment and two mucilage removal treatments. Significant differences were observed among treatments 8 weeks later, at the end of summer. In particular, dark filamentous algae were more abundant in all enriched plots, especially where mucilage and macroalgae had been removed; a higher percent cover of crustose coralline algae was instead observed where nutrients had been increased and no grazing pressure acted. Furthermore, the abundance of Dictyota spp. and Laurencia spp. was significantly higher in enriched mucilage-free plots where the grazing pressure was null or low. However, the effects of the treatments on the overall assemblage of the macroalgal community were not long persistent (36 weeks later). These results illustrate the capacity of a shallow-water macroalgal community to quickly recover from the simultaneous impacts of herbivory, nutrient enrichment, and mucilage

ClimateFish: A Collaborative Database to Track the Abundance of Selected Coastal Fish Species as Candidate Indicators of Climate Change in the Mediterranean Sea

Under the effects of global warming, many animals and plants are undergoing rapid distribution shifts. These changes can be particularly rapid in marine fishes, and many species have responded markedly to recent increases in sea temperature. ClimateFish is an open-access database, which collates abundance data for 7 Mediterranean indigenous and 8 non-indigenous fishes, proposed as candidate indicators of climate change. These species have been selected by a network of Mediterranean scientists based on their wide distribution, responsiveness to temperature conditions and easy identification. Data are periodically collected according to a standard visual census protocol in four different depth layers. At present, the database collates data on a total number of 101'771 observed individuals belonging to the 15 target species. Counts were realized along 3142 transects carried out in 7 Mediterranean countries between 2009 and 2021. This database, associated with climate data, offers new opportunities to investigate spatiotemporal effects of climate change and to test the effectiveness of each selected indicator. Data are available at https://doi.org/10.17882/86784.The Mediterranean ClimateFish initiative was initially conceived by the international basin wide monitoring program CIESM Tropical Signals (funded by the Albert II of Monaco Foundation) and subsequently supported by the Interreg Med Programme (Projects: MPA-ADAPT, grant number 1MED15_3.2_M2_337 and MPA Engage, grant number 5MED18_3.2_M23_007), 85% co funded by the European Regional Development Fund

The Management of the Beach-Cast Seagrass Wracks—A Numerical Modelling Approach

Seagrass wrack are commonly found on the beach face of the sandy shore all around the world and often persists in situ during the whole year, favouring the emergence of conflicts for the use of the sandy coasts for bathing or for other recreational purposes. As a consequence, these deposits are often removed from the beach during the summer months, temporary stocked, and relocated on the shore face in the next autumn or winter season. The selection of the sites on the shoreline where the leaves should be released before the storms season is often an issue, considering the optimization needs between the transportation costs and the oceanographic features of the dumping site. In this study, a numerical approach was proposed to identify the most suitable areas for the autumnal repositioning of the seagrass wracks for two beaches of Sardinia, an island located in the Western Mediterranean Sea where Posidonia oceanica (L. Delile, 1813) is the most widespread seagrass species. The method is based on the use of hydrodynamic, wave, and particle tracking models and provides important indications useful for the management of this type of practice that can be extended to all different type of beaches along the Mediterranean coasts