Trajectories of trace element accumulation in seagrass (Posidonia oceanica) over a decade reveal the footprint of fish farming.

To evaluate the effect of trace element (TE) release from fish farms on seagrass Posidonia oceanica, we compared TE concentrations (As, Cd, Co, Cu, Mn, Mo, Ni, Pb, V, Zn) in shoots near fish cages (Station 'Cage') with those away from them (Station 'Control') in two fish farm facilities (Site 1 and Site 2, North Aegean Sea, Greece). We assessed the present (i.e., 2021, year of sampling) and past (reconstructed period 2012-2020) accumulation of TEs using the living compartments (leaf blades, sheaths, rhizomes, roots, epiphytes) and the dead sheaths, respectively. We also assessed possible seagrass degradation by reconstructing past rhizome production. P. oceanica rhizome production at the 'Cage' stations was up to 50% lower than at the 'Control' stations. Most TE concentrations were higher at 'Cage' stations, but the differences often depended on the seagrass living compartment. Significant differentiation between 'Cage' and 'Control' stations was observed based on the TE concentrations of the dead sheaths during 2012-2020. The contamination level at the 'Cage' stations was mostly moderate in Site 1 and low in Site 2, during the reconstructed period, while an increasing contamination trend was found for certain potential phytotoxic TEs (As, Cu, Cd, Mo, V). Our results emphasize the need for the aquaculture industry to work towards a more ecologically aware approach.

Holothurians play an important role in mitigating the impacts of aquaculture on sediment conditions.

As deposit feeders contribute to bioremediation and nutrient recycling in sediments, positively impacting water and sediment quality, holothurians are candidate organisms for multitrophic aquaculture. This study aimed to investigate the potential of Holothuria poli to reduce the environmental footprint of fish farms through a benthocosm experiment. The experimental setup included four benthocosms with holothurians(H+) and four without (H-). The 58-day experiment included two phases: constant organic enrichment and recovery. In order to simulate the organic enrichment sediment conditions under a typical fish farm, a mixture of fish feces and fish feed pellets was added. Results showed that holothurians effectively reduced organic matter and H2S and increased redox, RPD depth and sediment oxygenation, thereby preventing anoxic conditions. Also, during the recovery phase, holothurians facilitated rapid sediment recovery, while the sediments without holothurians remained organic-enriched until the end of the experiment. The study emphasizes the significance of holothurians in mitigating the impacts of aquaculture on sediment conditions and so promoting environmental sustainability.

Transcriptomic Responses of a Lightly Calcified Echinoderm to Experimental Seawater Acidification and Warming during Early Development.

Ocean acidification (OA) and ocean warming (OW) are potential obstacles to the survival and growth of marine organisms, particularly those that rely on calcification. This study investigated the single and joint effects of OA and OW on sea cucumber Apostichopus japonicus larvae raised under combinations of two temperatures (19 °C or 22 °C) and two pCO2 levels (400 or 1000 µatm) that reflect the current and end-of-21st-century projected ocean scenarios. The investigation focused on assessing larval development and identifying differences in gene expression patterns at four crucial embryo-larval stages (blastula, gastrula, auricularia, and doliolaria) of sea cucumbers, using RNA-seq. Results showed the detrimental effect of OA on the early development and body growth of A. japonicus larvae and a reduction in the expression of genes associated with biomineralization, skeletogenesis, and ion homeostasis. This effect was particularly pronounced during the doliolaria stage, indicating the presence of bottlenecks in larval development at this transition phase between the larval and megalopa stages in response to OA. OW accelerated the larval development across four stages of A. japonicus, especially at the blastula and doliolaria stages, but resulted in a widespread upregulation of genes related to heat shock proteins, antioxidant defense, and immune response. Significantly, the negative effects of elevated pCO2 on the developmental process of larvae appeared to be mitigated when accompanied by increased temperatures at the expense of reduced immune resilience and increased system fragility. These findings suggest that alterations in gene expression within the larvae of A. japonicus provide a mechanism to adapt to stressors arising from a rapidly changing oceanic environment.

Contribution of holothuroids to the bioremediation and stimulation of the benthic microbial metabolism in aquaculture.

The implementation of sustainable system designs in aquaculture comprises the biological treatment of pollutants. In order to test the potential of Holothuria (Roweothuria) poli as a candidate for bioremediation of aquaculture wastes, the effect of this species on the benthic microbial activity was studied. Two experimental treatments, with and without holothuroids, were allocated in two benthocosms, each one containing four replicate tanks with undisturbed sediment and a volume of water above it. It was anticipated that H. poli could affect the overall assimilative capacity of the aquaculture effluent system either through the consumption of particulate organic matter or through the stimulation of benthic microbial metabolism. Indeed, the results indicated that the microbial activity related to the mineralization of the N and C was significantly different between the treatments verifying our hypothesis that deposit-feeders that move above or inside the sediment matrix can accelerate the depletion of organic matter through bioturbation.

Metals in tissues of marine fish from the Thermaikos Gulf, Eastern Mediterranean Sea: Detection of changes with trophic level.

Concentrations of 27 metals and elements were investigated in the tissues of 28 demersal and pelagic marine fish species from the Eastern Mediterranean Sea. The analysis indicated highest concentrations of accumulation for the majority of elements in the gills, liver, and scales; whereas, lowest assimilation of elements was reported in the muscle. Pelagic fish accumulated higher concentrations of elements than demersal fish. An inverse correlation due to the growth dilution effect was revealed among element concentrations in the gills, liver, muscle, and skin tissues, and fish size. Patterns of biomagnification for Cs solely were recorded in the muscle and patterns of biodilution for most elements were evident in the gills, liver, muscle, and skin tissues. A positive correlation was determined between trophic level and element concentrations in the scales which could not only be associated with fish diet, but also with metal concentrations in the ambient environment and scales size.

Viral Metagenomic Content Reflects Seawater Ecological Quality in the Coastal Zone.

Viruses interfere with their host's metabolism through the expression of auxiliary metabolic genes (AMGs) that, until now, are mostly studied under large physicochemical gradients. Here, we focus on coastal marine ecosystems and we sequence the viral metagenome (virome) of samples with discrete levels of human-driven disturbances. We aim to describe the relevance of viromics with respect to ecological quality status, defined by the classic seawater trophic index (TRIX). Neither viral (family level) nor bacterial (family level, based on 16S rRNA sequencing) community structure correlated with TRIX. AMGs involved in the Calvin and tricarboxylic acid cycles were found at stations with poor ecological quality, supporting viral lysis by modifying the host's energy supply. AMGs involved in "non-traditional" energy-production pathways (3HP, sulfur oxidation) were found irrespective of ecological quality, highlighting the importance of recognizing the prevalent metabolic paths and their intermediate byproducts. Various AMGs explained the variability between stations with poor vs. good ecological quality. Our study confirms the pivotal role of the virome content in ecosystem functioning, acting as a "pool" of available functions that may be transferred to the hosts. Further, it suggests that AMGs could be used as an ultra-sensitive metric of energy-production pathways with relevance in the vulnerable coastal zone and its ecological quality.

Assessing large-scale macrobenthic community shifts in the Aegean Sea using novel beta diversity modelling methods. Ramifications on environmental assessment.

The effects of large-scale environmental gradients on the spatial patterns of macrobenthic communities used in marine health assessment were evaluated using beta diversity methods. In this work, beta diversity partitioning and relevant ecological modelling methods revealed clear patterns between the northern and the southern parts of the Aegean Sea (Greece). A community turnover point was observed in the Evoikos Gulf marking the transition between the northern and the southern communities. The increased beta diversity in this work was due to species replacement driven mainly by the latitudinal gradients of bottom temperature and salinity whereas species richness did not present substantial differences between the Aegean macrobenthic communities. These findings are attributed to the ability of beta diversity methods to detect the environmental filtering that occurs in these marine provinces through the assessment of biotic interactions in respect to geographic distance. We propose a new standpoint of using beta diversity measures for benthic environmental assessment on a large spatial scale and in marine areas characterised by environmental gradients.

Prokaryotic and eukaryotic microbial community responses to N and P nutrient addition in oligotrophic Mediterranean coastal waters: Novel insights from DNA metabarcoding and network analysis.

The effects of the abrupt input of high quantities of dissolved inorganic nitrogen and phosphorus on prokaryotic and eukaryotic microbial plankton were investigated in an attempt to simulate the nutrient disturbances caused by eutrophication and climate change. Two nutrient levels were created through the addition of different quantities of dissolved nutrients in a mesocosm experiment. During the developed blooms, compositional differences were found within bacteria and microbial eukaryotes, and communities progressed towards species of faster metabolisms. Regarding the different nutrient concentrations, different microbial species were associated with each nutrient treatment and community changes spanned from the phylum to the operational taxonomic unit (OTU) level. Network analyses revealed important differences in the biotic connections developed: more competitive relationships were established in the more intense nutrient disturbance and networks of contrasting complexity were formed around species of different ecological strategies. This work highlights that sudden disturbances in water column chemistry lead to the development of entirely different microbial food webs with distinct ecological characteristics.

Modelling the effects of climate change on the distribution of benthic indicator species in the Eastern Mediterranean Sea.

The potential effects of climate change on the distribution of benthic species commonly used in marine ecological quality assessment were investigated using a spatial modelling approach. In this work, the relevance of the ecological groups that macrofaunal molluscs are assigned according to their sensitivity or tolerance to environmental disturbance was examined under the scope of the RCP 8.5 severe emissions scenario. The effects of climate change were more profound on species that are indicative of a specific suite of climatic conditions regarding temperature and salinity. Significant loss of habitat suitability was observed for the tolerant species Corbula gibba and Abra prismatica whereas the sensitive species Moerella donacina was least affected. In contrast, an overall expansion of the distributional potential was observed for the sensitive species Flexopecten hyalinus as newly suitable habitats are formed. As hypothesised, the current ecological grouping that depicts the sensitivity of a benthic species to an environmental stressor is irrelevant when assessing the effects of climate change. We propose a new standpoint of using benthic species as biotic tools based on their ecological niche requirements.

Can intensive fish farming for 20 years induce changes in benthic ecosystems on a scale of waterbody? An assessment from Cephalonia bay (Ionian Sea).

The environmental impacts of fish farming on benthic ecosystems beneath the fish cages have been widely addressed the past decades. However, the chronic release of nutrients can cause a shift in local primary productivity and a chronic increase in the sedimentation of organic material at a large spatial scale which could be reflected in benthic ecosystems. In this context, the indirect effects of aquaculture on the benthic ecosystem were studied in a semi-closed bay (Cephalonia, Ionian Sea) where a relatively large fish farm has been operating since 1982. Results from the present sampling were compared to historical data obtained in 1996 and 2001, in order to detect if nutrient release that could impact phytoplankton dynamics in the bay could indirectly alter benthic communities, as well. Macrofaunal communities have not shown deterioration but rather a small, yet statistically significant, improvement in diversity indices and ecological status indicators, and no significant change regarding bioturbation potential. This indicated that processes involved in nutrient consumption and transfer are highly effective in such an oligotrophic environment.

Molluscan indicator species and their potential use in ecological status assessment using species distribution modeling.

Marine habitat assessment using indicator species through Species Distribution Modeling (SDM) was investigated. The bivalves: Corbula gibba and Flexopecten hyalinus were the indicator species characterizing disturbed and undisturbed areas respectively in terms of chlorophyll a concentration in Greece. The habitat suitability maps of these species reflected the overall ecological status of the area. The C. gibba model successfully predicted the occurrence of this species in areas with increased physical disturbance driven by chlorophyll a concentration, whereas the habitat map for F. hyalinus showed an increased probability of occurrence in chlorophyll-poor areas, affected mainly by salinity. We advocate the use of C. gibba as a proxy for eutrophication and the incorporation of this species in monitoring studies through SDM methods. For the Mediterranean Sea we suggest the use of F. hyalinus in SDM as an indicator of environmental stability and a possible forecasting tool for salinity fluctuations.

Species-specific response to sulfide intrusion in native and exotic Mediterranean seagrasses under stress.

We explored the sulfur dynamics and the relationships between sediment sulfur and nutrient pools, seagrass structural and physiological variables and sulfide intrusion in native (Posidonia oceanica, Cymodocea nodosa) and exotic (Halophila stipulacea) Mediterranean seagrasses at six sites affected by cumulative anthropogenic pressures to understand the factors controlling sulfide intrusion in seagrass. Sensitive indicators of seagrass stress (leaf TN, δ15N, TS, Fsulfide) were increased at several sites, implying that seagrasses are under pressure. Sulfide intrusion was not related to sediment TOC but it was negatively related to shoot size and below-ground biomass. Sulfide intrusion in seagrass tissue was high in P. oceanica (12-17%) and considerably higher in C. nodosa (27-35%). Intrusion was particularly high in H. stipulacea (30-50%), suggesting that its possible biogeographical expansion due to warming of the Mediterranean may result in accumulation of sulfides in the sediments and hypoxia/anoxia with further implications in ecosystem function.

Benthic pelagic coupling in a mesocosm experiment: Delayed sediment responses and regime shifts.

A mesocosm experiment was performed to study benthic-pelagic coupling under a eutrophication gradient. Nine mesocosms were deployed in the facilities of the Hellenic Center for Marine Research in Crete, in the Eastern Mediterranean. The mesocosms were 4m deep, containing 1.5m3 of coastal water and, at the bottom, they included 85l of undisturbed sediment, collected from a semi-impacted area in the port of Heraklion, Crete. A eutrophication gradient was created by adding nutrients in the water column (Low and High) and the experiment lasted 58days. Water column and sediment environmental variables were measured at regular intervals. The results indicate that sedimentation caused by eutrophication in the water column affected sediment geochemical variables but in most cases a time lag was observed between the trophic status of the water column and the response of the sediment. Additionally, in the High eutrophication treatment, several fluctuations were observed and the system did not recover within the experimental duration, as opposed to the Low treatment which showed fewer fluctuations and signs of recovery.

Long-term (1998-2010) large-scale comparison of the ecological quality status of gulf of lions (NW Mediterranean) benthic habitats.

A comprehensive Mediterranean data set has been used to address 3 questions associated with the use of sensitivity/tolerance based biotic indices to infer the Ecological Quality status (EcoQs) of benthic habitats. Our results showed: (1) a significant effect of the reference database on derived sensitivity/tolerance measure (ES500.05) as well as associated Benthic Quality Index values and derived EcoQs; (2) a lack of correlation neither between BQI and AZTI Marine Biotic Index values nor between BQI and Multivariate-AZTI Marine Biotic Index values; (3) a lack of correlation between the values of the Benthic Habitat Quality Index (index derived from Sediment Profile Imagery) and those of either of the 3 tested biotic indices; and (4) a general agreement between the 3 tested biotic indices in describing the lack of global trend for the EcoQs of the Gulf of Lions despite the occurrence of significant changes in benthic macrofauna composition between 1998 and 2010.

One Step forward: Benthic Pelagic Coupling and Indicators for Environmental Status.

A large data set from the Eastern Mediterranean was analyzed to explore the relationship between seawater column variables and benthic community status. Our results showed a strong quantitative link between the seawater column variables (Chlorophyll a and Eutrophication Index) and various indicators describing benthic diversity and community composition. The percentage of benthic opportunistic species increased significantly in the stations with high trophic status of the seawater column and so did the strength of the coupling between values of seawater column and benthic indicators. The Eutrophication Index threshold level of 0.85, separating the "Bad and Poor" from "Moderate to High" conditions could serve as an acceptable critical value above which there is a readily observable change in benthic community composition.

Mussel farming in Maliakos Gulf and quality indicators of the marine environment: Good benthic below poor pelagic ecological status.

Biological and geochemical variables in the water column and sediments were monitored along a transect of a mussel farm located in a transitional environment in Maliakos Gulf, a semi-enclosed gulf in eastern Mediterranean. Analyses of water, sediment and macrofauna samples were used to calculate ecological status indicators in the context of the European Water Framework Directive. The water column ecological status was "Poor" or "Bad" showing little change with distance from the farm, but the ecological status of the benthic communities was found to be "Good," although there were quantitative changes in macrofaunal indices with distance from the farm.

The pattern of change in the abundances of specific bacterioplankton groups is consistent across different nutrient-enriched habitats in Crete.

A common source of disturbance for coastal aquatic habitats is nutrient enrichment through anthropogenic activities. Although the water column bacterioplankton communities in these environments have been characterized in some cases, changes in α-diversity and/or the abundances of specific taxonomic groups across enriched habitats remain unclear. Here, we investigated the bacterial community changes at three different nutrient-enriched and adjacent undisturbed habitats along the north coast of Crete, Greece: a fish farm, a closed bay within a town with low water renewal rates, and a city port where the level of nutrient enrichment and the trophic status of the habitat were different. Even though changes in α-diversity were different at each site, we observed across the sites a common change pattern accounting for most of the community variation for five of the most abundant bacterial groups: a decrease in the abundance of the Pelagibacteraceae and SAR86 and an increase in the abundance of the Alteromonadaceae, Rhodobacteraceae, and Cryomorphaceae in the impacted sites. The abundances of the groups that increased and decreased in the impacted sites were significantly correlated (positively and negatively, respectively) with the total heterotrophic bacterial counts and the concentrations of dissolved organic carbon and/or dissolved nitrogen and chlorophyll α, indicating that the common change pattern was associated with nutrient enrichment. Our results provide an in situ indication concerning the association of specific bacterioplankton groups with nutrient enrichment. These groups could potentially be used as indicators for nutrient enrichment if the pattern is confirmed over a broader spatial and temporal scale by future studies.

Metals and other elements in tissues of wild fish from fish farms and comparison with farmed species in sites with oxic and anoxic sediments.

Farmed fish and wild fish aggregating in the vicinity of four Mediterranean fish farms with different environmental conditions were sampled. Levels of metals (including As and Se) were measured in the muscle, liver, gills, bone and intestine. The wild fish from sites with anoxic substrata accumulate metals (including As and Se) from the ambient habitat in their gills whereas those from sites with oxic substrata concentrate these elements through their diet in their intestine. Tissues of wild fish aggregating around farm cages accumulate a greater number of these elements and with higher concentrations than farmed fish. Habitat, diet, ecological needs, fat content of fish, and protein expression may play an important role in these element differences between fish species. Fe in flathead grey mullet, As in surmullet, rainbow wrasse, grey gurnard and picarel and Hg in bogue may pose a risk for human health. Farmed and wild fish are good sources of P, K, Cr and Se while flathead grey mullet, picarel and comber are excellent sources of Ca and Se.

PAHs and n-alkanes in Mediterranean coastal marine sediments: aquaculture as a significant point source.

The occurrence of polycyclic aromatic and aliphatic hydrocarbons in fish feed, sediment trap material and marine sediments was examined at two fish farms in the eastern Mediterranean. The average (min-max) concentrations of polycyclic aromatic hydrocarbons (PAHs) in fish feed and particulate effluents were 316 (287-351) ng g(-1) DW and 487 (475-499) ng g(-1) DW, respectively. Lower PAH levels were determined in the underlying marine sediments. In the surface sediments under the farms (0 m distance from the edge of the cages) and in the immediate vicinity, the concentration levels of n-alkanes and PAHs were significantly higher than in the surrounding sediments in both sites. PAHs and n-alkanes individual component profiles of fish feed and sinking material were similar with the corresponding profiles of the sediment samples collected in the immediate vicinity around the cages. On a daily basis, the average PAH sedimentation fluxes under the cages was 24.4 microg m(-2) d(-1), which is considerably higher compared with the observed PAH sedimentary fluxes in the open eastern Mediterranean. Our results imply that fish farming is a significant source of these persistent organic pollutants (POPs) in the marine environment and therefore a likely change in the scale of production might introduce new sources of environmental risk. Further work is required in order to develop an appropriate monitoring system for the sustainable development of the aquaculture sector.

Effects of fish farming on the biological and geochemical properties of muddy and sandy sediments in the Mediterranean Sea.

The aim of this paper was to test how benthic fauna and biogeochemical properties of sediment will vary in response to similar levels of organic enrichment (induced by fish farming) as a function of bottom-habitat type (i.e., mud versus seagrass/coarse sediments), distance from the enrichment source and depth. Our results showed that samples from silty sediments in the vicinity of fish farms have higher TOC and TON values, higher oxygen consumption, higher PO(4) release and lower benthic diversity. In this context muddy sites are more likely to be identified as impacted/critical, than coarse sediment ones.