Structural and elemental analysis of the freshwater, low-Mg calcite coralline alga <i>pneophyllum cetinaensis</i>

Coralline algae are one of the most diversified groups of red algae and represent a major component of marine benthic habitats from the poles to the tropics. This group was believed to be exclusively marine until 2016, when the first freshwater coralline algae Pneophyllum cetinaensis was discovered in the Cetina River, southern Croatia. While several studies investigated the element compositions of marine coralline algal thalli, no information is yet available for the freshwater species. Using XRD, LA-ICP-MS and nano indentation, this study presents the first living low-Mg calcite coralline algae with Mg concentrations ten times lower than is common for the average marine species. Despite the lower Mg concentrations, hardness and elastic modulus (1.71 ± 1.58 GPa and 29.7 ± 18.0 GPa, respectively) are in the same range as other marine coralline algae, possibly due to other biogenic impurities. When compared to marine species, Ba/Ca values were unusually low, even though Ba concentrations are generally higher in rivers than in seawater. These low values might be linked to different physical and chemical characteristics of the Cetina River

First ex situ outplanting of the habitatforming seaweed Cystoseira amentacea var. stricta from a restoration perspective

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

Macrophytobenthic colonization of artificial reefs in the Ligurian Sea (Loano-Savona)

Lo studio della composizione del popolamento macrofitobentonico su strutture artificiali sommerse ha una storia decennale nella zona di Loano (Savona). Il presente lavoro mira a presentare le osservazioni preliminari di una nuova campagna in corso di svolgimento nella medesima zona. La flora e la struttura della comunità macrofitobentonica viene analizzata sulle strutture artificiali (piramidi) posizionate nella zona di Loano ad una profondità di 18 m. Il confronto con i dati storici presenti in letteratura fornirà un quadro dell'evoluzione del popolamento su tali substrati

Addressing reproductive stochasticity and grazing impacts in the restoration of a canopy-forming brown alga by implementing mitigation solutions

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 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 one was started in winter, after a marine heatwave that triggered early fertility, the second one in spring, when the species usually reproduces. 3. This study aimed to evaluate: 1) the disruptive effects of a thermal anomaly on the reproductive biology and performance in culture of T. barbata; and 2) the impact of the grazing pressure on juveniles after the outplanting. 4. The first cultivation was more productive than the second one, 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 T. 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 T. barbata. These stressors should be considered when developing plans to implement effective large-scale restoration of canopy-forming macroalgae

Microbe-assisted seedling crop improvement by a seaweed extract to address fucalean forest restoration

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

Rhodolith Beds Heterogeneity along the Apulian Continental Shelf (Mediterranean Sea)

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

Spatial patterns and drivers of benthic community structure on the northern Adriatic biogenic reefs

The northern Adriatic Sea (NAS) hosts numerous biogenic subtidal reefs that are considered biodiversity hotspots. Several studies have already investigated the origin and biodiversity of these reefs. However, many of them are still unexplored and further knowledge is needed for their conservation. Here, the spatial variability, epibenthic community structure, and environmental features that characterize these habitats were investigated. Fifteen randomly selected reefs were sampled between 2013 and 2017, including some remote sites that have never been studied before. A fuzzy k-means clustering method and redundancy analysis were used to find similarities among sites in terms of epibenthic assemblages and to model relationships with abiotic variables. The results showed that these reefs are highly heterogeneous in terms of species composition and geomorphological features. The results were also consistent with previous studies and highlighted three main types of benthic assemblages defined by the dominance of different organisms, mainly reflecting the coastal-offshore gradient: nearshore reefs, generally dominated by stress-tolerant species; reefs at a middle distance from the coast, characterized by sponges, non-calcareous encrusting algae and ascidians; offshore reefs, dominated by reef builders. However, distance from the coast was not the only factor affecting species distribution, as other local factors and environmental characteristics also played a role. This kind of biogenic reefs in temperate seas are still poorly known. The present work contributed to shed further light on these habitats, by complementing the results of previous studies on their natural diversity, highlighting the specificity of the epibenthic communities of NAS reefs and the need to improve current, still inadequate, conservation measures

NAMBER: A biotic index for assessing the ecological quality of mesophotic biogenic reefs in the northern Adriatic Sea

1. The aim of the present study was to propose a biotic index (North Adriatic Mesophotic BiogEnic Reefs, NAMBER) suitable for assessing the ecological quality of the mesophotic biogenic reefs of the northern Adriatic continental shelf based on photographic sampling. 2. At each of the 20 study sites, the degree of bioconstruction (expressed as percentage cover of crustose coralline algae), the α-diversity (expressed as the mean number of taxa), and the degree of sensitivity to human disturbance and climate change (based on literature data and expert judgement) of the benthic assemblages were selected as descriptors and combined in the NAMBER index, using the best values that the three metrics can currently achieve in the studied region as a reference. 3. The study highlighted that there was large spatial heterogeneity among reefs and high variability in the ecological quality values obtained by NAMBER, ranging from bad to high. The index indicates that reefs lying furthest from the coast, under substantially lower anthropogenic pressure, have a generally higher status of environmental quality. However, a clear geographical pattern did not emerge, as reefs close together often had different ecological qualities. 4. The NAMBER index, which combines three ecological descriptors, in accordance with the requirements of the European Marine Strategy Framework Directive, represents a specific adaptation to the northern Adriatic Sea of a multimetric index previously developed for the north-western Mediterranean Sea, capitalizing on previous knowledge and research efforts. 5. This multimetric biotic index provides an effective standardized tool for monitoring programmes and environmental impact assessments in the northern Adriatic mesophotic biogenic reefs and lays the foundation for ecosystem-based management and conservation in this basin

First freshwater coralline alga and the role of local features in a major biome transition

Coralline red algae are significant components of sea bottom and up to now considered as exclusively marine species. Here we presentthe first coralline alga from a freshwater environment, found in theCetina River (Adriatic Sea watershed).The alga is fully adapted to freshwater, as attested by reproductive structures, sporelings, and an inability to survive brackish conditions. Morphological and molecular phylogenetic analyses revealthe species belongs to Pneophyllum and is described as P. cetinaensis sp. nov.The marine-freshwatertransition most probably occurred during the last glaciation. The brackish-water ancestor was preadapted to osmotic stress and rapid changes in water salinity and temperature.The particular characteristics ofthe karst Cetina River, such as hard water enriched with dissolved calcium carbonate and a pH similarto the marine environment, favoured colonization ofthe river by a marine species.The upstream advance and dispersal is facilitated by exceptionally pronounced zoochory by freshwater gastropods. Pneophyllum cetinaensis defies the paradigm of Corallinales as an exclusively marine group