The Impact of Highly Weathered Oil from the Most Extensive Oil Spill in Tropical Oceans (Brazil) on the Microbiome of the Coral Mussismilia harttii.

In 2019, the largest oil spill ever recorded in tropical oceans in terms of extent occurred in Brazil. The oil from the spill was collected directly from the environment and used in an exposure experiment with the endangered reef-building coral Mussismilia harttii. The treatments of the experiment were control (without oil), 1% oil, 2.5% oil, and direct contact of coral with oil. The most abundant hydrocarbon in the seawater of the experiment was phenatrene, which is toxic to corals. However, overall, the concentration of PAHs was not very high. The analysis of the maximum photosynthetic capacity of Symbiodiniaceae dinoflagellates showed a small impact of oil on corals, mainly on the contact treatment. However, coral microbiomes were affected in all oil treatments, with the contact treatment showing the most pronounced impact. A greater number and abundance of stress-indicating and potentially pathogenic bacteria were found in all oil treatments. Finally, this highly weathered oil that had lain in the ocean for a long time was carrying potentially coral-pathogenic bacteria within the Vibrionaceae family and was able to transmit some of these bacteria to corals. Bacteria within Vibrionaceae are the main causes of disease in different species of corals and other marine organisms.

Consequences of prenatal exposure to contaminants in elasmobranchs: Biochemical outcomes during the embryonic development of Pseudobatos horkelii.

Coastal elasmobranchs are vulnerable to chemicals mostly due to their k-strategic life history characteristics and high trophic positions. Embryos might be particularly exposed through the maternal offloading of contaminants, possibly leading to disruptions during critical developmental phases. Yet, knowledge on biochemical outcomes of prenatal exposure in elasmobranchs is notably limited. Therefore, we aimed to investigate the effects of prenatal exposure to contaminants in embryos of the critically endangered Brazilian guitarfish, Pseudobatos horkelii. Polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), pharmaceuticals and personal care products, and metals were determined in embryos. Additionally, glutathione S-transferase activity (GST), glutathione (GSH), and metallothionein levels (MT) were analyzed. Finally, lipid peroxidation levels (LPO) and protein carbonyl groups (PCO) were assessed. Embryonic exposure depended on yolk consumption, which was conspicuous in earlier development. We observed a dilution effect of contaminants levels, potentially related to biotransformation of these compounds throughout the embryonic development. Nevertheless, GST was not correlated to contaminant concentrations. The multivariate relationship between antioxidant components (GSH and GST) and LPO and PCO was negative, suggesting the lack of efficient defense of these biomarkers in early development, leading to oxidative damage. In this context, our results indicate that prenatal exposure to contaminants might impact the redox status in embryos of P. horkelii, leading to oxidative damage. Furthermore, metal concentrations influenced MT levels, suggesting this as a potential detoxification pathway in this species.

Combined physiological and behavioral approaches as tools to evaluate environmental risk assessment of the water accommodated-fraction of diesel oil.

There is an increasing concern related to the toxic effects of the soluble portion of diesel oil on aquatic ecosystems and the organisms living in them. In this context, the aim of this study was to analyze the effects of diesel water accommodated-fraction (WAF) on behavioral and biochemical responses of mussels Perna perna. Animals were exposed to 5 and 20% of WAF for 96 h. Prior to the beginning of the experiments, Hall effect sensors and magnets were attached to the valves of the mussels. Valve gaping behavior was continuously recorded for 12 h of exposure and tissues (gills and digestive gland) were separated after 96 h of exposure. Overall, both behavior and biochemical biomarkers were altered due to WAF exposure. Animals exposed to WAF reduced the average amplitude of the valves and the fraction of time opened, and presented greater transition frequency, demonstrating avoidance behavior over the 12 h period. Furthermore, the biochemical biomarkers (GSH, GST, SOD and CAT) were altered following the 96 h of exposure to WAF. Considering the results presented, this study demonstrates the toxic potential of WAF in both shorter and longer exposure periods.

Health condition of Chelonia mydas from a foraging area affected by the tailings of a collapsed dam in southeast Brazil.

In 2015, the failure of the Fundão dam caused the release of 43 million m3 of tailings into the Doce River Basin, in southeast Brazil. It was considered the largest environmental disaster of the world mining industry. The tailings, composed mostly of heavy metals, caused massive destruction of the Doce River ecosystem endangering the organisms that live in the coastal zone where the mud reached the ocean. Among the exposed species are the sea turtles that use the region for food. The aim of this study was to evaluate the effect of contaminants on the health status of juvenile green sea turtles that feed in a coastal area exposed to ore mud (Santa Cruz) and to compare them with animals from an area not directly affected (Coroa Vermelha). A physical examination was performed to determine the health status. Blood samples were analyzed for hematological and biochemical parameters, and metal concentrations (As, Cd, Cr, Cu, Fe, Hg, Mn, Pb, and Zn). Santa Cruz sea turtles had more ectoparasites and a higher incidence of fibropapillomatosis. Statistically significant differences between sites were found for levels of calcium, phosphorus, glucose, protein, albumin, globulin, cholesterol, triglycerides, urea, CPK, ALT, and AST. The count of leukocytes, thrombocytes, and heterophils, as well as the concentrations of As and Cu were higher in Santa Cruz turtles. Together the results show a worse nutritional status and a greater degree of liver and kidney damage in animals affected by the tailings. The health status may indicate a physiological deficit that can affect their immune system and behavior, which is supported by the higher fibropapillomatosis tumor score and ectoparasite load in these animals. These results support the need for long-term monitoring of the exposed area to quantify the direct and indirect influence of the heavy metals levels on sea turtles and how this reflects the environmental health.

Metal Accumulation and Ion Regulation in the Fish Hyphessobrycon luetkenii Living in a Site Chronically Contaminated by Copper: Insights from Translocation Experiments.

Fish living in the João Dias creek (southern Brazil) have to deal with trace-metal contamination in the long-term basis, as this aquatic environment has been historically impacted by copper mining activities. In order to survive in this harsh environment, the local biota had to develop adaptations related to pollution tolerance. The aim of this study was to test if biochemical mechanisms related to osmoregulation were among these adaptations, using translocation experiments. Water ionic and trace-metal compositions were measured in a nonmetal impacted site (NMIS) and in a metal impacted site (MIS) of this creek. Also, whole-body metal accumulation, ion concentration and branchial enzyme activity (Na,K-ATPase and carbonic anhydrase) were evaluated in Hyphessobrycon luetkenii. In both NMIS and MIS, fish were collected and immediately stored, kept caged or translocated from sites. The result shows that waterborne Cu was 3.4-fold higher at the MIS. Accordingly, animals that had contact with this site showed elevated whole-body Cu levels. Moreover, both translocated groups showed elevated Na,K-ATPase activity. Additionally, fish translocated from the NMIS to the MIS showed lower carbonic anhydrase activity. These findings indicate that H. luetkenii chronically or acutely exposed to naturally elevated waterborne Cu showed a rapid Cu bioaccumulation but was unable to readily excrete it. Moreover, classic Cu osmoregulatory toxicity related to Na,K-ATPase inhibition was not observed. Conversely, impacts in ammonia excretion related to carbonic anhydrase inhibition may have occurred.

Response of biomarkers to metals, hydrocarbons and organochlorine pesticides contamination in crabs (Callinectes ornatus and C. bocourti) from two tropical estuaries (São José and São Marcos bays) of the Maranhão State (northeastern Brazil).

Response of biomarkers to chemical contamination was evaluated in crabs of the Callinectes genus (Callinectes ornatus and C. bocourti) from two tropical estuaries (São José and São Marcos bays) of the Maranhão State (northeastern Brazil). Biomarkers evaluated included hepatopancreatic metallothionein-like proteins (MTLP) and lipid peroxidation (LPO), as well as muscle acetylcholinesterase (AChE). Tissue concentrations of metals (pereiopod muscle and hepatopancreas), hydrocarbons (hepatopancreas) and organochlorine pesticides (hepatopancreas) were also evaluated. Crab samples were collected in three sites of each estuary (São Marcos Bay and São José Bay). Sampling was performed in August/2012 (dry season), January/2013 (rainy season), August/2013 (dry season), and January/2014 (rainy season). Concentrations of chemical contaminants and responses of biomarkers showed significant spatial (São Marcos Bay and São José Bay) and/or seasonal (dry and rainy seasons) and annual (2012-2014) variability. However, a general higher Zn concentration was observed in hepatopancreas of crabs from São José Bay. In turn, a general higher Cd concentration paralleled by oxidative damage (LPO) was observed in hepatopancreas of crabs from São Marcos Bay. As expected, these findings support the idea that this bay is more intensively or chronically impacted by industrial activities while the São José Bay is likely more affected by domestic activities. Interestingly, LPO level in crab hepatopancreas showed to be the most reliable and adequate biomarker to distinguish the two bays.

Aluminum bioconcentration in female Nile tilapia Oreochromis niloticus (Perciformes: Cichlidae) and the effects on pituitary gonadotropins.

In this study, we measured aluminum (Al) bioconcentration in the brain, ovaries, and liver of Oreochromis niloticus females, and analyzed the effects of exposure to Al and acidic pH on the gene expression of follicle-stimulating hormone (ßfsh) and luteinizing hormone (ßlh) in these animals. Mature females were divided into 4 groups, thus being maintained for 96 h in one of the following conditions: control at neutral pH (Ctr); Al at neutral pH (Al); acidic pH (Ac), and Al at acidic pH (Al-Ac). pH alone did not influence Al bioconcentration in the brain. The animals from the Al-Ac group bioconcentrated more Al in the ovaries than those from the Al group, while no differences were observed in the liver. Aluminum bioconcentration was higher in the brain than in the liver and ovaries in Al-exposed animals (Al and Al-Ac), and higher in the brain than in the ovaries in the Ctr and Ac groups. The liver bioconcentrates more Al than the ovaries in the females from the Ctr and Ac groups. Aluminum and/or acidic pH did not alter ßfsh gene expression, while ßlh gene expression decreased in females from the Al group. Aluminum acted as an endocrine disruptor, suggesting deleterious effects in reproduction that could result in ovulation failure. Aluminum can act directly and/or indirectly in the pituitary, affecting ovarian steroidogenesis and altering the reproductive endocrine axis of mature O. niloticus females in an acute period of exposure.

Elevated Temperature and Exposure to Copper Leads to Changes in the Antioxidant Defense System of the Reef-Building Coral Mussismilia harttii.

The frequency and severity of coral bleaching events have increased in recent years. Global warming and contamination are primarily responsible for triggering these responses in corals. Thus, the objective of this study was to evaluate the isolated and combined effects of elevated temperature and exposure to copper (Cu) on responses of the antioxidant defense system of coral Mussismilia harttii. In a marine mesocosm, fragments of the coral were exposed to three temperatures (25.0, 26.6, and 27.3°C) and three concentrations of Cu (2.9, 5.4, and 8.6 µg/L) for up to 12 days. Levels of reduced glutathione (GSH) and the activity of enzymes, such as superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and glutamate cysteine ligase (GCL), were evaluated on the corals and symbionts. The short exposure to isolated and combined stressors caused a reduction in GSH levels and inhibition of the activity of antioxidant enzymes. After prolonged exposure, the combination of stressors continued to reduce GSH levels and SOD, CAT, and GCL activity in symbionts and GST activity in host corals. GCL activity was the parameter most affected by stressors, remaining inhibited after 12-days exposure. Interesting that long-term exposure to stressors stimulated antioxidant defense proteins in M. harttii, demonstrating a counteracting response that may beneficiate the oxidative state. These results, combined with other studies already published suggest that the antioxidant system should be further studied in order to understand the mechanisms of tolerance of South Atlantic reefs.

Arsenic, lead and cadmium concentrations in caudal crests of the yacare caiman (Caiman yacare) from Brazilian Pantanal.

Wetlands are environments of extreme importance due to their high biodiversity and invaluable ecosystem services. Nevertheless, wetlands worldwide are under the increasing threat of the effects of contaminants, which put at risk the biota and the ecosystems. Herein the concentrations of non-essential and toxic elements arsenic (As), lead (Pb) and cadmium (Cd) in caudal crests of the yacare caiman (Caiman yacare) from the Brazilian Pantanal, one of the largest wetlands in the world, were investigated aiming to compare concentrations in individuals from a large pristine area (Southern Pantanal) to animals from an area close to potential sources of anthropogenic emissions (Northern Pantanal). Levels above the limit of detection were found for all the elements in the majority of the analysed samples. The highest mean concentration was detected for As in samples from both sites, followed by Pb and Cd in samples from Southern Pantanal, and Cd and Pb in samples from Northern Pantanal. Significant negative correlations were found between As and Cd concentrations and the size of the individuals from the Southern Pantanal. Surprisingly, concentrations of all three elements were higher in pristine Southern Pantanal, but with significant differences only for Cd. This result suggests that natural sources and processes may be acting for the mobilisation and availability of As, Pb and Cd for Southern Pantanal biota, which reflected in the contamination of C. yacare. Data from the literature showed higher levels of As, Pb and Cd in abiotic compartments in Southern Pantanal and adjacent plateaus in comparison to Northern Pantanal, corroborating the higher concentrations detected in biotic samples from the south. Thus, natural sources seem to play a relevant role in the contamination of biota by As, Pb and Cd in Southern Pantanal, characterising an intriguing pattern that could be tested for other species.

Effects of sublethal Cd, Zn, and mixture exposures on antioxidant defense and oxidative stress parameters in early life stages of the purple sea urchin Strongylocentrotus purpuratus.

Oxidative stress parameters were evaluated during the first 72 h of embryonic development of purple sea urchin Strongylocentrotus purpuratus continuously exposed to control conditions, to cadmium alone (Cd, 30 µg/L), to zinc alone (Zn, 9 µg/L) or to a Cd (28 µg/L) plus Zn (9 µg/L) mixture. These sublethal concentrations represent ∼ 10% of the acute EC50. Bioaccumulation, antioxidant capacity against peroxyl radicals (ACAP), total glutathione (GSH) level, glutathione-S-transferase (GST), glucose-6-phosphate dehydrogenase (G6PDH) and superoxide dismutase (SOD) activity, and lipid peroxidation (LPO) were analyzed at 24 h (blastula), 48 h (gastrula), and 72 h (pluteus) stages of development. Zinc (an essential metal) was well-regulated, whereas Cd (non-essential) bioaccumulated and whole-body [Cd] increased from blastula to pluteus stage in sea urchin larvae. In controls, ACAP progressively declined from 24 h to 72 h, while LPO reciprocally increased, but other parameters did not change. Cd alone was more potent than Zn alone as a pro-oxidant, with the major effects being decreases in SOD activity and parallel increases in LPO throughout development; GST activity also increased at 24 h. Zn alone caused only biphasic disturbances of ACAP. In all cases, the simultaneous presence of the other metal prevented the effects, and there was no instance where the oxidative stress response in the presence of the Cd/Zn mixture was greater than in the presence of either Cd or Zn alone. Therefore the sublethal effects of joint exposures were always less than additive or even protective, in agreement with classical toxicity data. Furthermore, our results indicate that SOD and Zn can play important roles in protecting sea urchin embryos against Cd-induced lipid peroxidation.

Energy metabolism enzymes inhibition by the combined effects of increasing temperature and copper exposure in the coral Mussismilia harttii.

The combined effects of exposure to increasing temperature and copper (Cu) concentrations were evaluated in the zooxanthellate scleractinian coral Mussismilia harttii. Endpoints analyzed included activity of enzymes involved in glycolysis (pyruvate kinase, PK; lactate dehydrogenase, LDH), Krebs cycle (citrate synthase, CS; isocitrate dehydrogenase; IDH), electron transport chain (electron transport system, ETS) and pentose phosphate pathway (glucose-6-phosphate dehydrogenase, G6PDH). Coral polyps were kept under control conditions (25.0 ±â€¯0.1 °C; 2.9 ±â€¯0.7 µg/L Cu) or exposed to combined treatments of increasing temperature (26.6 ±â€¯0.1 °C and 27.3 ±â€¯0.1 °C) and concentrations of dissolved Cu (5.4 ±â€¯0.9 and 8.6 ±â€¯0.3 µg/L) for 4 and 12 days using a mesocosm system. PK activity was not affected by stressors. LDH, CS, IDH, ETS and G6PDH activities were temporally inhibited by stressors alone. CS, ETS and G6PDH activities remained inhibited by the combination of stressors after 12 days. Furthermore, all combinations between increasing temperature and exposure Cu were synergistic after prolonged exposure. Taken together, stressors applied alone led to temporary inhibitory effects on energy metabolism enzymes of the coral M. harttii, however, prolonged exposure reveals strong deleterious effects over the metabolism of corals due to the combination of stressors. The present study is the first one to give insights into the combined effects of increasing temperature and Cu exposure in the energy metabolism enzymes of a scleractinian coral. Findings suggest that moderate Cu contamination in future increasing temperature scenarios can be worrying for aerobic and oxidative metabolism of M. harttii.

Life-time exposure to waterborne copper II: Patterns of tissue accumulation and gene expression of the metal-transport proteins ctr1 and atp7b in the killifish Poecilia vivipara.

The involvement of transporting proteins on copper (Cu) bioaccumulation was evaluated in the killifish Poecilia vivipara chronically exposed to environmentally relevant concentrations of waterborne Cu. Fish (<24 h-old) were maintained under control condition or exposed to different waterborne Cu concentrations (5, 9 and 20 µg/L) for 28 and 345 days in saltwater. Following exposure periods, Cu accumulation and the expression of genes encoding for the high affinity Cu-transporter (ctr1) and the P-type Cu-ATPase (atp7b) were evaluated. Whole-body metal accumulation and gene expression were evaluated in fish exposed to 28 days. Similarly, in fish exposed to 345 days, liver, gills and gut were also evaluated. No fish survival was observed after exposure to 20 µg/L for 345 days. Whole-body Cu accumulation was significantly higher in fish exposed to 20 µg/L Cu for 28 days and in fish exposed to 9 µg/L for 345 days in comparison to control animals. Similarly, tissue Cu accumulation was significantly higher in fish exposed to 9 µg/L for 345 days in comparison to control animal. However, no significant accumulation was observed in fish muscle. Following exposure for 28 days, whole-body ctr1 expression was slightly induced in fish exposed to 9 µg/L. In turn, no significant change in ctr1 expression was observed following exposure to Cu for 345 days. Differently, whole-body atp7b expression was markedly up-regulated in the whole-body of fish exposed Cu for 28 days and in tissues of fish exposed to Cu for 345 days. These findings indicate the expression of atp7b is more responsive to Cu accumulation in P. vivipara than ctr1 expression and, therefore, more suitable to be used as a biomarker of exposure to this metal. Also, we argue that the expression of atp7b is sustained at elevated levels for as much time as fish are maintained in Cu contaminated water.

Disturbance in Na+ regulation in cells rich in mitochondria isolated from gills of the yellow clam Mesodesma mactroides exposed to copper under different osmotic conditions.

Cells rich in mitochondria were isolated from gills of the seawater clam Mesodesma mactroides, incubated in isosmotic saline solution (840 mOsmol/kg H2O), and exposed (3 h) to environmentally realistic Cu concentrations (nominally: 0, 5, 9 and 20 µg/L). In cells exposed to 20 µg Cu/L, Cu accumulation, Na+ content reduction and carbonic anhydrase (CA) activity inhibition were observed, without significant changes in cell viability and Na+,K+-ATPase (NKA) activity. In the absence of Cu, cell viability and Cu content were reduced in hyposmotic media respect with the control, without changes in Na+ content and enzyme (CA and NKA) activities. In the presence of 5 or 9 µg/L Cu, cell Cu content was increased, especially at 670 mOsmol/kg H2O. Cell Na+ content and NKA activity were reduced after exposure to 20 µg/L Cu at 670 mOsmol/kg H2O. In turn, CA activity was dependent on Cu concentration, being significantly reduced in cells exposed to 9 and 20 µg/L Cu in both hyposmotic conditions. These findings indicate that Cu also negatively affects Na+ regulation in gill cells of the seawater clam M. mactroides, with Cu toxicity increasing at hyposmotic conditions. Also, they indicate that physiology is more important than water chemistry in predicting Cu toxicity in environments of changing salinity, pointing out CA activity as a potential biomarker of Cu exposure.

Metal accumulation and expression of genes encoding for metallothionein and copper transporters in a chronically exposed wild population of the fish Hyphessobrycon luetkenii.

In the present study, metal (As, Cd, Cu, Fe, Mn, Pb and Zn) accumulation and expression of genes involved in metal metabolism (metallothioneins, ATP7A and CTR1) were evaluated in gills and muscle of the fish Hyphessobrycon luetkenii living in the João Dias creek, a site historically (~1870-1996) contaminated with a metal mixture associated with copper mining (Minas do Camaquã, southern Brazil). Fish were collected in a metal impacted site of the João Dias creek and kept in a cage at this site (PP fish) or translocated to a non-metal impacted reach of this creek (PC fish). Gill metal concentrations and metallothionein gene expression were lower in PC fish than in PP fish at any experimental time (24, 48 and 72 h). In muscle, no significant changes were observed. These findings indicate that metal accumulated in gills of wild fish chronically exposed to the metal mixture are more easily excreted than those accumulated in the muscle. In this case, expression of gene encoding for metallothionein is shown to play a key role in the regulation of metal accumulation in gills of H. luetkenii living in an area historically contaminated with a metal mixture associated with copper mining.

Cadmium in tissues of green turtles (Chelonia mydas): A global perspective for marine biota.

Cadmium (Cd) is a metal of toxicological interest because of its potential high toxicity to organisms and ability to biomagnify. Evaluating concentrations of Cd in organisms on a large spatial scale can provide insights to its global distribution. This study examined Cd concentrations in kidney and liver tissues of 137 specimens of green turtles (Chelonia mydas) collected in Australia, Brazil, Hawaii, Japan, and the continental United States (Gulf of Mexico). We used comparative analyses of kidney and liver of 35 individuals, of which seven turtles from each locality belong to the same size class for comparison purposes between their ocean of origin. Cd was detected in all samples, with the highest bioconcentration in kidneys. Specimens originating from the Pacific Ocean had significantly higher mean Cd levels in liver (13.24 µg/g) and kidney (34.17 µg/g) than the specimens collected in the Atlantic Ocean with lower mean values in liver (1.00 µg/g) and kidney (4.04 µg/g). Furthermore, Cd concentrations in turtle tissues were generally greater than concentrations found in other marine organisms, for example dolphins. This result was unexpected because dolphins occupy a higher trophic level than green turtles which are only herbivorous. A possible explanation is a change in feeding habits of green turtles, in which juveniles feed in near shore habitats, potentially resulting in greater Cd accumulation in juveniles compared to adults. This global distribution trend has also been observed in other marine organisms (e.g., insects, birds, and mammals) and indicates that global factors may be more important than regional factors in determining Cd concentrations of marine organisms. CAPSULE: Global factors are more relevant than local factors in the distribution of cadmium in biota, using green turtle as a sentinel species.

Biochemical and physiological effects of nickel in the euryhaline crab Neohelice granulata (Dana, 1851) acclimated to different salinities.

The estuarine crab Neohelice granulata was maintained under control condition or exposed to sublethal concentrations of dissolved Ni (measured: 128 and 1010µg/L) for 96h at different salinities (2 and 30ppt). After metal exposure, whole-body oxygen consumption was measured and tissue (hemolymph, gills, hepatopancreas and muscle) samples were collected. Control crabs acclimated to 2ppt salinity showed lower hemolymph concentrations of Na+ (33%), Mg2+ (19%) and K+ (30%), as well as increased LPO levels in anterior gills (379%), posterior gills (457%) and hepatopancreas (35%) with respect to those acclimated to 30ppt salinity. In crabs acclimated to 2ppt salinity, Ni exposure increased whole-body oxygen consumption (75%), hemolymph K+ concentration (52%), hemolymph (135%) and hepatopancreas (62%) LDH activity. Also, it reduced hemolymph Cl- concentration (16%) and muscle LDH activity (33%). In crabs acclimated to 30ppt salinity, Ni exposure increased LDH activity in hemolymph (195%), hepatopancreas (126%) and muscle (53%), as well as hemolymph osmolality (10%), Cl- (26%) and Ca2+ (20%) concentration. It also reduced hepatopancreas lipid peroxidation (20%) and hemolymph Mg2+ (29%) and K+ (31%) concentration. These findings indicate that N. granulata is hyper-osmoregulating in 2ppt salinity and hypo-regulating in 30ppt salinity, showing adjustments of hemolymph ionic composition and metabolic rates, with consequent higher oxidative damage to lipids in low salinity (2ppt). Ni effects are associated with metabolic (aerobic and anaerobic) disturbances in crabs acclimated to 2ppt salinity, while osmotic and ionoregulatory disturbances were more evident in crabs acclimated to 30ppt salinity.

Contrasting effects of a classic Nrf2 activator, tert-butylhydroquinone, on the glutathione-related antioxidant defenses in Pacific oysters, Crassostrea gigas.

Nrf2 is a well-known transcription factor controlling a number of antioxidant defense-related genes, which is understudied in bivalves. In this study, oysters Crassostrea gigas were exposed for 24, 48 and 96 h to 10 or 30 µM tert-butylhydroquinone (tBHQ), a classic Nrf2 activator. At 96 h, a clear induction of GSH-related antioxidant defenses was observed in gills of tBHQ-exposed animals, including GSH, glutathione S-transferase (GST), glutathione peroxidase (GPx) and glutathione reductase (GR). Unexpectedly, the activities of GST, GPx and GR were significantly decreased 24 h after tBHQ treatment, suggesting a possible inhibition, which was supported by in vitro experiments. GR mRNA (24 h) and protein levels (24 and 96 h) were increased by tBHQ treatment, confirming its induction, possibly by the Nrf2 pathway. The conserved domains at C. gigas Keap1 and Nrf2 proteins and the clear induction of GSH-related antioxidant defenses by tBHQ, a classical Nrf2 inducer, support the idea of a functional Nrf2/Keap1 pathway in bivalves. tBHQ also proved to be a tool to explore redox regulatory mechanisms in bivalves.

Copper effects on biomarkers associated with photosynthesis, oxidative status and calcification in the Brazilian coral Mussismilia harttii (Scleractinia, Mussidae).

Seawater contamination with metals, such as copper (Cu), is a notable local impact threatening coral reefs. Cu effects on biomarkers associated with photosynthesis, oxidative status and calcification were evaluated in the Brazilian coral Mussismilia harttii using a marine mesocosm facility. Polyps were kept under control conditions (1.9 µg L-1 Cu) or exposed to dissolved Cu (3.0, 4.8, and 6.7 µg L-1) for 12 days. Photochemical efficiency of the photosystem II of symbiotic algae (zooxanthellae) was measured and polyps were analyzed for antioxidant capacity, lipid peroxidation, DNA damage, and carbonic anhydrase Ca-ATPase, Mg-ATPase and (Ca,Mg)-ATPase activities after 12 days. Results highlighted the effects of Cu exposure, leading corals to an oxidative stress condition [increased total antioxidant capacity (TAC) and DNA damage] and a possible reduced calcification ability [decreased (Ca,Mg)-ATPase activity]. Therefore, biomarkers associated with oxidative status (TAC and DNA damage) and calcification [(Ca, Mg)-ATPase] are indicated as good predictors of corals health.

Effects of increasing temperature on antioxidant defense system and oxidative stress parameters in the Antarctic fish Notothenia coriiceps and Notothenia rossii.

Antioxidant defense system (ADS) and oxidative stress parameters were evaluated in the Antarctic fish Notothenia rossii and N. coriiceps exposed to increasing temperature. Acclimated fish were kept at 0°C or exposed to 4°C for 1day (N. rossii) or to 2 and 4°C for 1 and 6 days (N. coriiceps). Measurements were assessed in brain, gills, liver, white muscle and erythrocytes. Parameters analyzed included antioxidant capacity against peroxyl radicals (ACAP); reduced glutathione (GSH) and metallothionein-like proteins (MTLP) concentration; superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and glutamate-cysteine ligase (GCL) activity; lipid peroxidation (LPO) level and protein carbonyl (PC) concentration. Increased liver GST activity was observed in N. rossii exposed to 4°C for 1day. Increased muscle GPx activity was observed in N. coriiceps after exposure to 2°C for 1day. Reduced gill GPx activity and increased liver SOD activity were observed after exposure to 4°C for 1day. In N. coriiceps, increased gill GCL activity and reduced gill GPx activity, as well as reduced liver MTLP were observed after exposure to 2°C for 6 days. Reduced brain SOD activity and increased brain LPO; reduced gill ACAP, GSH concentration and GPx activity, as well as increased gill GCL activity; reduced liver ACAP, MTLP, SOD activity, GST activity and increased liver and erythrocytes LPO were observed after exposure to 4°C for 6 days. These findings indicate that ADS is more responsive to short-term increasing temperature in the sluggish N. coriiceps than in the active N. rossii. However, responses of N. coriiceps to long-term increasing temperature were transient and did not prevent tissue oxidative damage. Considering the predicted increase in temperature in the Southern Ocean over the next decades, our findings suggest that Antarctic fishes are sensitive to ocean warming, displaying tissue oxidative damage associated with the thermal stress.

Antioxidant defense system and oxidative status in Antarctic fishes: The sluggish rockcod Notothenia coriiceps versus the active marbled notothen Notothenia rossii.

Adaptive responses of antioxidant defense systems (ADS) to changes in increased levels of activity are critical, especially in Antarctic fishes. The benthopelagic marbled notothen (Notothenia rossii) shows higher spontaneous activity than the benthonic and sluggish rockcod (N. coriiceps). Therefore, we hypothesize that species-related responses of ADS would occur to counteract different rates of reactive oxygen species formation in these two Antarctic fish. Here we evaluated ADS and oxidative damage in tissues (brain, gills, liver and white muscle) of the two Antarctic fish. Despite no significant differences in lipid and protein oxidative damage were observed, we actually found species- and tissue-specific differences in ADS. Gill metallothionein-like proteins (MTLP) and liver reduced glutathione (GSH) concentrations were higher in N. coriiceps than in N. rossii. Brain and gill antioxidant capacity against peroxyl radicals (ACAP); gill enzyme [glutamate-cysteine ligase (GSL), superoxide dismutase (SOD) and catalase (CAT)] activity; liver GCL and SOD activity; and white muscle CAT activity were higher in N. rossii than in N. coriiceps. Therefore, the more active fish (N. rossii) maintains higher activities of enzymes involved in superoxide ions (O2.-) detoxification and GSH production in peripheral tissues (gills, liver and white muscle). This allows the more active fish (N. rossii) to keep levels of lipid and protein oxidative damage similar to those observed in the sluggish fish (N. coriiceps). It is worth noting that the more active fish also shows a higher brain antioxidant capacity, which could involve other non-enzymatic antioxidants like vitamins C and E. In contrast, N. coriiceps shows lower consumption of non-enzymatic antioxidants in peripheral tissues than N. coriiceps. As hypothesized, our results indicate that differences in ADS profiles between fish species are likely related to their habits and metabolic rates. This would imply in different fish abilities to deal with oxidative stress associated with increasing seawater temperature.