Marine mesocosm system: A reliable tool for testing bioaccumulation and effects of seawater enrichment with dissolved iron in reef organisms.

In 2015, a marine mesocosm facility was designed and implemented by the Coral Vivo Project in its research station (Porto Seguro, Bahia State, Brazil) to initially study the effects of global impacts, especially ocean warming and acidification, on coral reefs. However, local impacts, including seawater contamination with metal(loid)s, are considered as a major threat to coral reefs. Also, in 2015, the largest disaster involving a mining dam occurred in Brazil. Iron (Fe) mining tailings originated from the dam failure affected not only freshwater ecosystems (rivers, lakes and lagoons), but also adjacent beaches, mangroves, restingas, reefs and other marine systems. Seawater, sediments and biota were contaminated with metal(loid)s, especially Fe, arsenic (As), mercury (Hg) and manganese (Mn). Therefore, we aimed to adapt the marine mesocosm facility of the Coral Vivo Project to evaluate the bioaccumulation and biological impacts of increasing concentrations of dissolved Fe on a diversity of reef organisms. Results obtained indicate a great versatility and reliability of the marine mesocosm system for application in biological and ecological studies on the isolated effect of seawater dissolved Fe on reef organisms of different functional groups simultaneously.•Studies involving seawater enrichment with dissolved Fe can be performed using a marine mesocosm system.•The marine mesocosm is a reliable tool to study the isolated effects of metal(loid)s on reef organisms.

Sex-biased response of pollution biomarkers in fish: Insights from the killifish Poecilia vivipara.

Copper ions (Cu) are one of the most frequent trace-contaminants found in Brazilian waters and, although considered as an essential element, in high concentrations can accumulate and induce toxicity. Biomarkers are important tools that can be used to assess these impacts, but to be considered trustworthy, they have to be previously tested in target organisms through laboratory studies under controlled conditions. However, many of these experiments are conducted using only males, as it is believed that the hormonal variation of females can bias the results, increasing data variability. Notwithstanding, few studies have actually tested this hypothesis, highlighting the importance of considering and measuring the role of sex in ecotoxicological studies. The aim this study was to evaluate the influence of sex on biomarkers classically used in environmental monitoring programs using the fish Poecilia vivipara as model. For this, females and males were exposed for 96 h to two Cu concentrations (9 and 20 µg/L) and a control group. In liver and gills, Cu accumulation, total antioxidant capacity (TAC) and lipid peroxidation (LPO) were evaluated. In addition, samples of peripheral blood were used for neutrophil to lymphocyte ratio determination, a measure of the onset of secondary stress. Results show that Cu hepatic accumulation did not differ between females and males, but higher levels of this metal were observed in exposed animals compared to control fish. Additionally, interactive effects were observed for hepatic LPO, as males showed elevated oxidative damage in comparison to females. Moreover, Cu exposure elevated hepatic LPO relative to control only in males, but this increase in oxidative damage was not accompanied by changes in liver TAC. On the other hand, differences in branchial Cu accumulation and LPO were not observed. Conversely, control females showed elevated TAC in comparison to control males, but Cu exposure eliminated this difference. Cu exposure also induced an increase in the N:L ratio, indicating the presence of a secondary stress response unrelated to sex. Ultimately, the findings of this study demonstrate that sex can influence the response of biomarkers that are typically used in ecotoxicological investigations in a multifaceted manner. As a result, using animals from a singular sex in such studies may result in consequential outcomes, potentially leading to underestimation or overestimation of results.

Peroxynitrite Generation and Increased Heterotrophic Capacity Are Linked to the Disruption of the Coral-Dinoflagellate Symbiosis in a Scleractinian and Hydrocoral Species.

Ocean warming is one of the greatest global threats to coral reef ecosystems; it leads to the disruption of the coral-dinoflagellate symbiosis (bleaching) and to nutrient starvation, because corals mostly rely on autotrophy (i.e., the supply of photosynthates from the dinoflagellate symbionts) for their energy requirements. Although coral bleaching has been well studied, the early warning signs of bleaching, as well as the capacity of corals to shift from autotrophy to heterotrophy, are still under investigation. In this study, we evaluated the bleaching occurrence of the scleractinian coral Mussismillia harttii and the hydrocoral Millepora alcicornis during a natural thermal stress event, under the 2015-2016 El Niño influence in three reef sites of the South Atlantic. We focused on the link between peroxynitrite (ONOO-) generation and coral bleaching, as ONOO- has been very poorly investigated in corals and never during a natural bleaching event. We also investigated the natural trophic plasticity of the two corals through the use of new lipid biomarkers. The results obtained first demonstrate that ONOO- is linked to the onset and intensity of bleaching in both scleractinian corals and hydrocorals. Indeed, ONOO- concentrations were correlated with bleaching intensity, with the highest levels preceding the highest bleaching intensity. The time lag between bleaching and ONOO- peak was, however, species-specific. In addition, we observed that elevated temperatures forced heterotrophy in scleractinian corals, as Mu. harttii presented high heterotrophic activity 15 to 30 days prior bleaching occurrence. On the contrary, a lower heterotrophic activity was monitored for the hydrocoral Mi. alicornis, which also experienced higher bleaching levels compared to Mu. hartii. Overall, we showed that the levels of ONOO- in coral tissue, combined to the heterotrophic capacity, are two good proxies explaining the intensity of coral bleaching.

Carbonic anhydrase activity as a potential biomarker for acute exposure to copper in corals.

Coral reefs are subjected to climate change and are severely impacted by human activities, with copper (Cu) being a relevant physiological stressor for corals at local scale. The ecological relevance of parameters measured at biochemical or cellular level is now considered an extremely important feature in environmental studies, and can be used as early warning signs of environmental degradation. In this context, the effects of acute exposure (96 h) to Cu were assessed on the maximum photochemical efficiency of zooxanthellae (Fv/Fm) and on the activity of key enzymes [carbonic anhydrase (CA) and Ca-ATPase] involved in coral physiology using the scleractinian coral Mussismilia harttii as a biological model. Corals were exposed to different concentrations of dissolved Cu (4.6-19.4 µg/L) using two different experimental approaches: a laboratory closed system and a marine mesocosm system. Fv/Fm values and Ca - ATPase activity were not affect by exposure to Cu in any of the exposure systems. However, a significant reduction in CA activity was observed in corals exposed to 11.9 and 19.4 µg Cu/L in the laboratory and at all concentrations of Cu tested in the mesocosm system (4.6, 6.0 and 8.5 µg/L). Based on the sensitivity of this enzyme to the short period of exposure to sublethal concentrations of Cu in both experimental approaches, the present study suggests the use of CA activity as a potential biomarker to be used in biomarker-based environmental monitoring programs in coral reefs.

Effects of increasing temperature alone and combined with copper exposure on biochemical and physiological parameters in the zooxanthellate scleractinian coral Mussismilia harttii.

Effects of increasing temperature alone and in combination with exposure to dissolved copper (Cu) were evaluated in the zooxanthellate scleractinian coral Mussismilia harttii using a marine mesocosm system. Endpoints analyzed included parameters involved in metabolism [maximum photosynthetic capacity of zooxanthellae (Fv/Fm), chlorophyll a and ATP concentrations], calcification [carbonic anhydrase (CA) and Ca2+-Mg2+-ATPase activity], and oxidative status [antioxidant capacity against peroxyl radicals (ACAP) and lipid peroxidation (LPO)]. Coral polyps were collected, acclimated and exposed to three increasing temperature conditions [25.0±0.1°C (control; average temperature of local seawater), 26.6±0.1°C and 27.3±0.1°C] using a marine mesocosm system. They were tested alone and in combination with four environmentally relevant concentrations of dissolved Cu in seawater [2.9±0.7 (control; average concentration in local seawater), 3.8±0.8, 5.4±0.9 and 8.6±0.3µg/L] for 4, 8 and 12days. Fv/Fm reduced over the experimental period with increasing temperature. Combination of increasing temperature with Cu exposure enhanced this effect. CA and Ca2+-Mg2+-ATPase activities increased up to 8days of exposure, but recovered back after 12days of experiment. Short-term exposure to increasing temperature or long-term exposure to the combination of stressors reduced LPO, suggesting the occurrence of a remodeling process in the lipid composition of biological membranes. ACAP, ATP and chlorophyll a were not significantly affected by the stressors. These findings indicate that increasing temperature combined with exposure to dissolved Cu increase susceptibility to bleaching and reduce growth in the zooxanthellate scleractinian coral M. harttii.