Transcriptomic profiles and diagnostic biomarkers in the Mediterranean seagrasses Posidonia oceanica and Cymodocea nodosa reveal mechanistic insights of adaptative strategies upon desalination brine stress.

Seawater desalination by reverse osmosis is growing exponentially due to water scarcity. Byproducts of this process (e.g. brines), are generally discharged directly into the coastal ecosystem, causing detrimental effects, on benthic organisms. Understanding the cellular stress response of these organisms (biomarkers), could be crucial for establishing appropriate salinity thresholds for discharged brines. Early stress biomarkers can serve as valuable tools for monitoring the health status of brine-impacted organisms, enabling the prediction of long-term irreversible damage caused by the desalination industry. In this study, we conducted laboratory-controlled experiments to assess cellular and molecular biomarkers against brine exposure in two salinity-sensitive Mediterranean seagrasses: Posidonia oceanica and Cymodocea nodosa. Treatments involved exposure to 39, 41, and 43 psu, for 6 h and 7 days. Results indicated that photosynthetic performance remained unaffected across all treatments. However, under 43 psu, P. oceanica and C. nodosa exhibited lipid oxidative damage, which occurred earlier in P. oceanica. Additionally, P. oceanica displayed an antioxidant response at higher salinities by accumulating phenolic compounds within 6 h and ascorbate within 7 d; whereas for C. nodosa the predominant antioxidant mechanisms were phenolic compounds accumulation and total radical scavenging activity, which was evident after 7 d of brines exposure. Finally, transcriptomic analyses in P. oceanica exposed to 43 psu for 7 days revealed a poor up-regulation of genes associated with brassinosteroid response and abiotic stress response, while a high down-regulation of genes related to primary metabolism was detected. In C. nodosa, up-regulated genes were involved in DNA repair, cell cycle regulation, and reproduction, while down-regulated genes were mainly associated with photosynthesis and ribosome assembly. Overall, these findings suggest that 43 psu is a critical salinity-damage threshold for both seagrasses; and despite the moderate overexpression of several transcripts that could confer salt tolerance, genes involved in essential biological processes were severely downregulated.

Desalination brine effects beyond excess salinity: Unravelling specific stress signaling and tolerance responses in the seagrass Posidonia oceanica.

Desalination has been proposed as a global strategy for tackling freshwater shortage in the climate change era. However, there is a concern regarding the environmental effects of high salinity brines discharged from desalination plants on benthic communities. In this context, seagrasses such as the Mediterranean endemic and ecologically important Posidonia oceanica have shown high vulnerability to elevated salinities. Most ecotoxicological studies regarding desalination effects are based on salinity increments using artificial sea salts, although it has been postulated that certain additives within the industrial process of desalination may exacerbate a negative impact beyond just the increased salinities of the brine. To assess the potential effect of whole effluent brines on P. oceanica, mesocosm experiments were conducted within 10 days, simulating salinity increment with either artificial sea salts or brines from a desalination plant (at 43 psµ, 6 psµ over the natural 37 psµ). Morphometrical (growth and necrosis), photochemical (PSII chlorophyll a fluorometry), metabolic, such as hydrogen peroxide (H2O2), thiobarbituric reactive substances (TBARS) and ascorbate/dehydroascorbate (ASC/DHA), and molecular (expression of key tolerance genes) responses were analyzed in each different treatment. Although with a still positive leaf growth, associated parameters decreased similarly for both artificial sea salt and brine treatments. Photochemical parameters did not show general patterns, although only P. oceanica under brines demonstrated greater energy release through heat (NPQ). Lipid peroxidation and upregulation of genes related to oxidative stress (GR, MnSOD, and FeSOD) or ion exclusion (SOS3 and AKT2/3) were similarly incremented on both hypersalinity treatments. Conversely, the ASC/DHA ratio was significantly lower, and the expression of SOS1, CAT, and STRK1 was increased under brine influence. This study revealed that although metabolic and photochemical differences occurred under both hypersalinity treatments, growth (the last sign of physiological detriment) was similarly compromised, suggesting that the potential effects of desalination are mainly caused by brine-associated salinities and are not particularly related to other industrial additives.

A risk assessment on Zostera chilensis, the last relict of marine angiosperms in the South-East Pacific Ocean, due to the development of the desalination industry in Chile.

Seagrasses, which are considered among the most ecologically valuable and endangered coastal ecosystems, have a narrowly limited distribution in the south-east Pacific, where Zostera chilensis is the only remaining relict. Due to water scarcity, desalination industry has grown in the last decades in the central-north coasts of Chile, which may be relevant to address in terms of potential impacts on benthic communities due to their associated high-salinity brine discharges to subtidal ecosystems. In this work, we assessed ecophysiological and cellular responses to desalination-extrapolable hypersalinity conditions on Z. chilensis. Mesocosms experiments were performed for 10 days, where plants were exposed to 3 different salinity treatments: 34 psu (control), 37 psu and 40 psu. Photosynthetic performance, H2O2 accumulation, and ascorbate content (reduced and oxidized) were measured, as well as relative gene expression of enzymes related to osmotic regulation and oxidative stress; these, at 1, 3, 6 and 10 days. Z. chilensis showed a decrease in photosynthetic parameters such as electron transport rate (ETRmax) and saturation irradiance (EkETR) under hypersalinity treatments, while non-photochemical quenching (NPQmax) presented an initial increment and a subsequent decline at 40 psu. H2O2 levels increased with hypersalinity, while ascorbate and dehydroascorbate only increased under 37 psu, although decreased along the experimental period. Increased salinities also triggered the expression of genes related to ion transport and osmolyte syntheses, but salinity-dependent up-regulated genes were mostly those related to the reactive oxygen species metabolism. The relict seagrass Z. chilensis has shown to withstand increased salinities that may be extrapolable to desalination effects in the short-term. As the latter is not fully clear in the long-term, and considering the restricted distribution and ecological importance, direct brine discharges to Z. chilensis meadows may not be recommended.

Posidonia oceanica L. (Delile) meadows regression: Long-term affection may be induced by multiple impacts.

Coastal development has an undeniable impact on marine ecosystems resulting in the detriment of the more sensible communities. Posidonia oceanica meadows are climax communities which offer a wide variety of ecosystem services both ecological and socio-economic. Human-derived impact on these habitats has been widely assessed although conclusions may vary depending on the area. P. oceanica meadow regression next to the city of Alicante (SE Spain) was analyzed on the long term (1984-2014) using bionomic cartographies and side-scan sonar images and, during the last two decades (2003-2021), using cover percentage and shoot density descriptors in the remaining meadow. Results showed a 25% colonized area reduction since 1984, this process being more rapid during the 1984-1994 period and decreasing with time. Cover and density have suffered a significant decrease in the last 20 years, mainly in the upper limit of the meadow. Dead matte cover was also assessed and have shown a significant increase in the same period following an inverse trend with the other metrics. There are several coastal impacts which have co-occurred in the area in the last few decades (port enlargement, brine and sewage discharges, industrial activity) thus resulting in the regression of the meadow. The existing negative trend of the measured descriptors indicate the necessity of implementing management actions which focus on the present sources of impact and actively reduce their effect on P. oceanica beds.