Temporal changes in metal and arsenic concentrations in blood and feathers of tropical seabirds after one of the largest environmental disasters associated with mining.

Monitoring of contaminant levels in wildlife over time is a tool for assessing the presence and persistence of environmental impacts at ecosystem, community and population levels. Tropical seabirds breeding in the Abrolhos Archipelago, 70 km off the Brazilian coast, forage in areas under the influence of the Doce River discharge. In 2015, the Fundão Dam collapsed and released ca 60 million tons of iron ore tailings into the ocean. In the present study, red-billed tropicbirds Phaethon aethereus and brown boobies Sula leucogaster breeding in Abrolhos were monitored over four years (2019-2022) for metal (Fe, Mn, Zn, Cu, Cr, Hg, Pb, Cd) and metalloid (As) concentrations in blood and feathers. Over six sampling events, metal (loid) concentrations showed strong temporal variation in both tissues. Overall, feathers showed greater element concentrations than blood, with stronger correlations between elements, especially Mn and the nonessential As, Cd, Hg and Pb. Mn is one of the major chemical markers of the Fundão Dam tailings. Metal (loid) concentrations in the tropical seabirds evaluated were above suggested threshold levels for most nonessential elements (As, Cd and Pb), especially in February 2021, when metal (loid) concentrations peaked in feathers. In this case, values were orders of magnitude higher than those observed in other sampling events. This occurred one year after a major rainy season in the Doce River basin, which increased river discharge of contaminated mud into the ocean, where contaminants are further remobilized by winds and currents, resulting in transference through the marine food web. This finding is consistent to what has been observed for other ecosystem compartments monitored in the region under the influence of the Doce River. Our findings highlight the utility of using tropical seabirds as sentinels of marine pollution, revealing strong temporal patterns in metal (loid) concentrations associated to bottom-up climatic processes.

Potency and specificity of amiloride and its analogues on branchial sodium fluxes in freshwater trout and goldfish.

There is a consensus that electroneutral Na+/H+ exchangers (NHEs) are important in branchial Na+ uptake in freshwater fish. There is also widespread belief, based on mammalian data, that EIPA [5-(N-ethyl-N-isopropyl)-amiloride]], and HMA [5-(N,N-hexamethylene)-amiloride)] are more potent and specific in blocking Na+ uptake than amiloride. We evaluated this idea by testing the three drugs at 10-7 to 10-4 M, i.e. 0.1 to 100 µM in two model species, rainbow trout (Oncorhynchus mykiss) and goldfish (Carassius auratus), using 22Na+ to measure unidirectional Na+ influx and efflux rates. In both species, the potency order for inhibiting unidirectional Na+ influx was HMA > amiloride > EIPA (IC50 values in the 10-70 µM range), very different from in mammals. At 100 µM, all three drugs inhibited Na+ influx by >90% in both species, except for amiloride in goldfish (65%). However, at 60-100 µM, all three drugs also stimulated unidirectional Na+ efflux rates, indicating non-specific effects. In trout, HMA and EIPA caused significant increases (2.1- to 2.3-fold) in efflux rates, whereas in goldfish, significant efflux elevations were greater (3.1- to 7.2-fold) with all three drugs. We conclude that the inhibitory potency profile established in mammals does not apply to the NHEs in fish gills, that non-specific effects on Na+ efflux rates are a serious concern, and that EIPA and HMA offer no clear benefits in terms of potency or specificity. Considering its much lower cost, we recommend amiloride as the drug of choice for in vivo experiments on freshwater fishes.

Validation of qPCR reference genes in the endangered annual killifish Austrolebias charrua considering different tissues, gender and environmental conditions.

The annual killifish Austrolebias charrua is an endangered species, endemic to the southern region of South America, which inhabits temporary ponds that emerges in the rainy season. The main anthropogenic threat driving the extinction of A. charrua stems from extensive agriculture, primarily due to the widrespread use of glyphosate-based herbicides near their habitats. Annual killifishes have been used as models for ecotoxicological studies but, up to now, there are no studies about reference genes in any Austrolebias species. This represents an obstacle to the use of qPCR-based technologies, the standard method for gene expression quantification. The present study aimed to select and validate potential reference genes for qPCR normalization in the annual killifish Austrolebias charrua considering different tissues, gender and environmental conditions. The candidate reference genes 18 s, actb, gapdh, ef1a, shox, eif3g, and the control gene atp1a1 were evaluated in male and female individuals in three different tissues (brain, liver, and gills) under two experimental conditions (control and acute exposition to Roundup Transorb®). The collected tissues were submitted to RNA extraction, followed by cDNA synthesis, cloning, sequencing, and qPCR. Overall, 18 s was the most stable reference gene, and 18 s and ef1a were the most stable combination. Otherwise, considering all variables, gapdh and shox were the least stable candidate genes. Foremost, suitable reference genes were validated in A. charrua, facilitating accurate mRNA quantification in this species, which might be useful for developing molecular tools of ecotoxicological assessment based on gene expression analysis for environmental monitoring of annual killifish.

A Pilot Study of Gene Expression Modulation from Antioxidant System of Killifish Austrolebias charrua After Exposure to Roundup Transorb®.

Roundup Transorb® (RDT) is the most popular glyphosate-based herbicide (GHB) used in agriculture, and its impact extends to non-target organisms. The annual killifish Austrolebias charrua is an endangered species endemic to southern South America and inhabits temporary ponds. This study evaluates the effects of RDT concentrations (0.065 and 5 mg/L GAE) on A. charrua exposed for 96 h. Gene expression of cat, sod2, gstα, gclc, and ucp1 was evaluated on the liver and gills. Highlighting that even at low concentrations permitted by Brazilian legislation, the RDT can have adverse effects on A. charrua.

The microbial profile of rivers and lagoons three years after the impact of the world's largest mining disaster (Fundão dam, Brazil).

The collapse of the Fundão tailings dam (Minas Gerais, Brazil) was the largest environmental disaster in Brazil's history and in the world mining industry. This disaster carried approximately 55 million m3 of iron ore tailings along the rivers and the lagoons of the Doce river basin. Although multiple studies assessed the impact on microbial communities in those rivers and lagoons right after the dam rupture, it is not known whether the microbiome in those environments remains impacted years after the disaster. Assessing the microbiome is very important to evaluate impacts and evaluate the health of the environment, due to the several ecological roles played by microorganisms. Here, we evaluated the impact of the dam failure on water and sediment bacteriome and archaeome by high-throughput next-generation sequencing. Samples were taken from two rivers and six lagoons during the dry and rainy seasons approximately three years post disturbance. The results showed a large number and abundance of microbial groups associated with the presence of heavy metals and mine tailings sediments. Some of these microorganisms were also reported in large abundance in the impacted rivers shortly after the Fundão dam rupture. Among the most abundant microorganisms in the Doce River, we can highlight the bacteria hgcI clade and the archaea Nitrososphera sp. in the water, and the bacteria Anaerolineaceae sp. in the sediment. These results suggest that the microbiome of the rivers and the lagoons in the Doce river basin remains severely impacted by the Fundão tailings dam failure even three years after the disaster. The presence of those microorganisms can also help to assess the occurrence of the Fundão dam sediment in other environments.

Oxidative Stress in a Wild Population of the Freshwater Fish Hyphessobrycon Luetkenii Chronically Exposed to a Copper Mining Area: New Insights into Copper Toxicology.

In this study, we examine markers of oxidative stress in the tetra Hyphessobrycon luetkenii collected from two locations in the copper contaminated João Dias creek (southern Brazil). Also, specimens were translocated from a clean reference section of the creek to a polluted stretch and vice-versa. Fish were held at in submerged cages for 96 h and then sacrificed. Nuclear abnormalities in erythrocytes and total antioxidant capacity, lipid peroxidation and protein carbonylation in gills, brain, liver and muscle displayed similar trends in both groups. Lipid peroxidation increased in all tissues of individuals translocated to the polluted site but only in liver and muscle of those translocated to the reference site. Increased protein carbonylation was also observed in gills of individuals translocated to the reference location. These results suggest similar oxidative stress among fish from the reference and polluted locations and that long-term metals exposure may require adaptations toward oxidative stress responses.

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.

The bioaccumulation of waterborne zinc in tissues of silver catfish (Rhamdia quelen) and its effect on biochemical parameters.

Silver catfish (Rhamdia quelen) is a fish species with neotropical distribution, and is a potential model organism to study polluted environment. The aim of this study is to analyze the response of silver catfish to environmental concentrations of waterborne zinc (Zn) over 96 h. Significant metal accumulation was seen in gill, intestine and liver tissues. No significant accumulation was seen in muscle tissue. Lipid peroxidation increased in the brain, and decreased in the muscle and liver at all levels of exposure. Zinc exposure led to decreased protein carbonyl levels in the brain and increased levels in the liver. The activity of catalase in the liver was reduced for all exposed groups. Glutathione S-transferase activity decreased in the brain at the highest level of exposure and in the liver at all Zn concentrations tested. Non-protein thiols increased in the muscle and in the gills after exposure. Ascorbic acid levels increased in the brain and in the gills. Exposure to Zn also altered the metabolic parameters, causing decreased lactate and ammonia levels in the muscle, and decreased glycogen in the liver. Zinc exposure increased ammonia and amino acid levels in the liver, and increase glycogen and amino acid levels in muscle tissue. Our results demonstrate that exposure to environmentally relevant concentrations of Zn led to accumulation of metals in the tissues of silver catfish, with significant changes in biochemical parameters.

Transcriptional effects in the estuarine guppy Poecilia vivipara exposed to sanitary sewage in laboratory and in situ.

The urban growth has increased sanitary sewage discharges in coastal ecosystems, negatively affecting the aquatic biota. Mangroves, one of the most human-affected coastal biomes, are areas for reproduction and nursing of several species. In order to evaluate the effects of sanitary sewage effluents in mangrove species, this study assessed the hepatic transcriptional responses of guppy fish Poecilia vivipara exposed to sanitary sewage 33% (v:v), using suppressive subtraction hybridization (SSH), high throughput sequencing of RNA (Ion-proton) and quantification of transcript levels by qPCR of some identified genes in fish kept in a sewage-contaminated environment. Genes identified are related predominantly to xenobiotic biotransformation, immune system and sexual differentiation. The qPCR results confirmed the induction of cytochrome P450 1A (CYP1A), glutathione S transferase A-like (GST A-like) methyltransferase (MET) and UDP glycosyltransferase 1A (UDPGT1A), and repression of complement component C3 (C3), doublesex and mab-3 related transcription factor 1 (DMRT1), and transferrin (TF) in the laboratory experiment. In the field exposure, the transcript levels of CYP1A, DMRT1, MET, GST A-like and UDPGT1A were higher in fishes exposed at the contaminated sites compared to the reference site. Chemical analysis in fish from the laboratory and in situ experiments, and surface sediment from the sewage-contaminated sites revealed relevant levels of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyl (PCBs) and linear alkylbenzenes (LABs). These data reinforce the use of P. vivipara as a sentinel for monitoring environmental contamination in coastal regions.

Carbonic Anhydrase as a Biomarker of Global and Local Impacts: Insights from Calcifying Animals.

The emission of greenhouse gases has grown in unprecedented levels since the beginning of the industrial era. As a result, global climate changes, such as heightened global temperature and ocean acidification, are expected to negatively impact populations. Similarly, industrial and urban unsustainable development are also expected to impose local impacts of their own, such as environmental pollution with organic and inorganic chemicals. As an answer, biomarkers can be used in environmental programs to assess these impacts. These tools are based in the quantification of biochemical and cellular responses of target species that are known to respond in a sensitive and specific way to such stresses. In this context, carbonic anhydrase has shown to be a promising biomarker candidate for the assessment of global and local impacts in biomonitoring programs, especially in marine zones, such as coral reefs, considering the pivotal role of this enzyme in the calcification process. Therefore, the aim of this review is to show the recent advances in the carbonic anhydrase research and the reasons why it can be considered as a promising biomarker to be used for calcifying organisms.

Roundup® Herbicide Decreases Quality Parameters of Spermatozoa of Silversides Odontesthes Humensis.

The silverside (Odontesthes humensis) is a very interesting model for toxicological studies due its high sensitivity and need for good water quality. The aim of this study was to evaluate the effects of Roundup on spermatozoa of O. humensis, after acute exposure. The fish were exposed to 0 and 7.8 mg L-1 (a.e.) of glyphosate, respectively. Through computer-assisted sperm analysis, a significant decrease in concentration, total and progressive motility, average path distance, straight line distance, path average velocity, curved line velocity, straight line velocity linearity, wobble, amplitude of lateral head displacement, cross beat frequency, and motility period of silverside spermatozoa exposed to Roundup was observed. Also, increase in membrane fluidity, ROS production and lipid peroxidation and a decrease in the mitochondrial functionality was observed in spermatozoa of Roundup exposed silversides. It was demonstrated that Roundup exposure in a concentration that can be achieve in natural water bodies soon after its application in fields is able to cause losses in several sperm quality parameters, consequently decreasing the fertilization potential of O. humensis spermatozoa.

Effects of Experimental Lead Exposure on Testis of the Chestnut Capped Blackbird Chrysomus ruficapillus.

Lead (Pb) effects on testis histology, as well as sperm quality and oxidative status were evaluated in male Chestnut Capped Blackbird (Chrysomus ruficapillus). Wild blackbirds were captured, immediately sampled (field group) or kept in captivity and treated with a single intraperitoneal injection of saline solution (control) or saline solution with Pb acetate (50 or 100 mg/kg Pb). Seven days after injection, whole blood, ductus deferens and testis samples were collected. Increased Pb concentrations were observed in whole blood and testis of Pb-exposed blackbirds with respect to those from field and control blackbirds. Sperm cells of Pb-exposed blackbirds showed loss of membrane integrity, mitochondrial functionality, and DNA integrity. Also, oxidative damage was observed in testis of blackbirds injected with 100 mg/kg Pb. These findings indicate that Pb is accumulated in testis of C. ruficapillus, inducing severe morphological and biochemical injury that can compromise the reproductive performance of male blackbirds. Although the exposure scenario (Pb acetate, high dosage and intraperitoneal injection) tested in the present study would likely not occur in the wild, it was adequate to show potential and relevant toxic effects of Pb in wild birds.

The Effects of Acute Copper and Ammonia Challenges on Ammonia and Urea Excretion by the Blue Crab Callinectes sapidus.

Copper (Cu) is a persistent environmental contaminant that elicits several physiological disturbances in aquatic organisms, including a disruption in ammonia regulation. We hypothesized that exposure to Cu in a model crustacean (blue crab, Callinectes sapidus) acclimated to brackish water (2 ppt) would lead to hyperammonemia by stimulating an increase in ammonia production and/or by inhibiting ammonia excretion. We further hypothesized that urea production would represent an ammonia detoxification strategy in response to Cu. In a pilot experiment, exposure to 0, 100, and 200 µg/L Cu for 6 h caused significant concentration-dependent increases in ammonia excretion (J amm). Based on these results, an acute 24-h 100 µg/L Cu exposure was conducted and this similarly caused an overall stimulation of J amm during the 24-h period, indicative of an increase in ammonia production. Terminal haemolymph total ammonia content (T amm) was unchanged, suggesting that while ammonia production was increased, there was no inhibition of the excretion mechanism. In support of our second hypothesis, urea excretion (J urea) increased in response to Cu exposure; haemolymph [urea] was unaffected. This suggested that urea production also was increased. To further test the hypothesis that J urea increased to prevent hyperammonemia during Cu exposure, crabs were exposed to high environmental ammonia (HEA; 2.5 mmol/L NH4HCO3) for 12 h in a separate experiment. This led to a fourfold increase in haemolymph T amm, whereas J urea increased only transiently and haemolymph [urea] was unchanged, indicating that urea production likely does not contribute to the attenuation of hyperammonemia in blue crabs. Overall, Cu exposure in blue crabs led to increased ammonia and urea production, which were both eliminated by excretion. These results may have important implications in aquaculture systems where crabs may be exposed to elevated Cu and/or ammonia.

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.

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.

Effects of sodium chloride exposure on ion regulation in larvae (glochidia) of the freshwater mussel Lampsilis fasciola.

The salinization of freshwater can have negative effects on ecosystem health, with heightened effects in salt-sensitive biota such as glochidia, the larvae of freshwater mussels. However, the toxicological mechanism underlying this sensitivity is unknown. Therefore, Lampsilis fasciola glochidia were exposed to NaCl (nominally 0.25 and 1.0 g/L) prepared in reconstituted moderately-hard water (control), as well as to a dilution of that water (1:4) with ultrapure reference water (diluted control). Unidirectional Na(+) influx (measured with (22)Na) was evaluated after 1, 3 and 48 h of exposure. In addition, unidirectional Cl(-) influx (measured with (36)Cl), whole-body ion (Cl(-) and Na(+)) concentrations, and glochidia viability (measured as the ability to close valves) were assessed after 48 h of exposure. Significantly reduced glochidia viability (56%) was observed after exposure to 1.0 g/L NaCl. Na(+) influx was significantly higher in glochidia exposed to both 0.25 and 1.0 g/L NaCl for 1h than in those kept under control conditions. After 3 and 48 h of exposure, differences in Na(+) influx rate between salt-exposed and control glochidia were generally reduced, indicating that larvae may be able to, at least temporarily, recover their ability to regulate Na(+) influx when exposed to elevated NaCl concentration. Compared to the moderately-hard water control, whole-body Na(+) and Cl(-) concentrations were relatively unchanged in glochidia exposed to 0.25 g/L NaCl, but were significantly elevated in glochidia exposed to 1.0 g/L NaCl and the diluted control. While Na(+) influx rate had recovered to the control level after 48 h of exposure to 1.0 g/L NaCl, Cl(-) influx rate remained elevated, being ~7-fold higher than the Na(+) influx rate. These findings suggest that the loss of viability observed when glochidia were exposed to a high NaCl concentration (1.0 g/L) could be caused by ionoregulatory disturbances mainly associated with an elevated Cl(-) influx.

Responses of biomarkers in wild freshwater mussels chronically exposed to complex contaminant mixtures.

Subcellular biochemical biomarkers are valuable early warning indicators of environmental contaminant effects. Thus, the present study evaluated several biomarkers and the relationships among them in wild freshwater mussels (Lasmigona costata) from a gradient of metal exposure and differential levels of other urban-related influences in the Grand River (ON, Canada). The biomarkers examined are related to metal exposure [gill ion and metal concentrations (Na, K, Ca, Mg, Cd, Cu, Ni, Pb and Zn)], oxidative status [reactive oxygen species (ROS), catalase (CAT), superoxide dismutase (SOD), antioxidant capacity (ACAP)], sulfhydryl (SH) metabolism [glutathione (GSH), protein sulfhydryl groups (SH protein), glutathione S-transferase (GST), glutathione reductase (GR)], and lipid peroxidation. Gill metal concentration increased proportionally to waterborne metal concentration and disturbances in osmotic and divalent cations (Ca and Mg) concentrations were observed. This suggests that the observed effects are associated with metal exposure, although simultaneous relationships with other contaminants are also possible. Oxidative status biomarkers (ROS, SOD, CAT and ACAP) were more sensitive to urban-influences than gill metal concentration. In contrast, biomarkers involving SH metabolism (GSH, SH protein, total SH, GR and GST) were more correlated with gill metal concentration. Oxidative damage occurred when both metal and urban-related influences were high. Mechanistically, the way of dealing with oxidative stress changed when mussels were exposed to high levels of contaminants. The reduction in ROS content, SOD and CAT activity, and ACAP accompanying the stimulation of detoxification metabolism via SH (GSH and SH protein contents, GST and GR activities) and their association with gill metal concentration are discussed.

The effects of copper and nickel on the embryonic life stages of the purple sea urchin (Strongylocentrotus purpuratus).

The aim of this research was to generate data on the mechanisms of toxicity of copper [Cu (4-12 µg/L)] and nickel [Ni (33-40 µg/L)] during continuous sublethal exposure in seawater (32 ppt, 15 °C) in a sensitive test organism (Strongylocentrotus purpuratus) at its most sensitive life stage (developing embryo). Whole-body ions [calcium (Ca), sodium (Na), potassium (K), and magnesium (Mg)], metal burdens, Ca uptake, and Ca ATPase activity were measured every 12 h during the first 72-84 h of development. Ionoregulatory disruption was clearly an important mechanism of toxicity for both metals and occurred with minimal metal bioaccumulation. Most noteworthy was a significant disruption of Ca homeostasis, which was evident from an inhibition of unidirectional Ca uptake rates, whole-body Ca accumulation, and Ca ATPase activity intermittently during 72-84 h of development. At various times, Cu- and Ni-exposed embryos also displayed lower levels of K and increased levels of Na suggesting inhibition of Na/K ATPase activity. Greater levels of Mg during initial stages of development in Cu-exposed embryos were also observed and were considered a possible compensatory mechanism for disruptions to Ca homeostasis because both of these ions are important constituents of the developing spicule. Notably, most of these effects occurred during the initial stages of development but were reversed by 72-84 h. We therefore propose that it is of value to study the toxic impacts of contaminants periodically during development before the traditional end point of 48-72 h.

Arsenic and metal levels in snake tissues from Lagoa Santa Karst, Brazil.

Concentrations of one metalloid (As) and eight metals (Cd, Cr, Cu, Hg, Mn, Ni, Pb, and Zn) were determined in tissues (muscle, liver, and kidney) of eight snake species (Bothrops neuwiedi, Crotalus durissus, Dipsas mikanii, Epicrates crassus, Helicops modestus, Micrurus carvalhoi, Oxyrhopus guibei, and Oxyrhopus trigeminus) from Lagoa Santa Karst. Except for Cu and Zn, all other analyzed elements were detected in concentrations within the ranges previously reported for snakes inhabiting polluted areas, emphasizing Hg (specific Hg mean concentrations varied from 0.87 to 9.76 µg g-1 d.w). The highest mean concentrations of all elements except Zn were found in muscle samples of the false corals O. guibei (means ranged from 2.01 [Pb] to 9.76 [Hg]). The highest Zn mean concentration (13.77 µg g-1 d.w) was detected in the kidney of the water snake H. modestus. No significant correlation was found between element concentrations and body size for all species. Significant interorgan differences were observed for As, Cr, Cu, Hg, Mn, Pb, and Zn concentrations in the three tissues in H. modestus. Significant interspecific differences were found in at least one organ for all elements. Significant pairwise differences were found between diet specialist species and between these species and broader diet species, while no significant difference was found between the broader diet species. The bioaccumulation of As and metals in snakes from Lagoa Santa Karst could be associated with natural rock dissolution and erosion processes but also with the wide-scale mining in the region and the increased agriculture and urbanization.

Distribution of copper in the Atlantic and Pacific Oceans using green turtles (Chelonia mydas) as a bioindicator.

Marine pollution by trace elements is a global concern due to potential toxicity to species and ecosystems. Copper is a fundamental trace element for many organisms; however, it becomes toxic at certain concentrations. The green turtle (Chelonia mydas) is a good sentinel species, due to its circumglobal distribution, long life cycle, coastal habits when juvenile, and is subject to environmental pollution. Quantifying and comparing copper levels makes it possible to understand the availability of this trace element in nature. During this research, comparisons were made between the levels of copper found in the liver, kidneys, and muscles of 35 turtles, from the United States (Hawaii and Texas), Brazil, and Japan. Copper was found in all specimens. In the liver, animals from Hawaii (91.08 µg g-1), Texas (46.11 µg g-1), and Japan (65.18 µg g-1) had statistically equal means, while those from Brazil (16. 79 µg g-1) had the lowest means. For the kidney, copper means were statistically equal for all Hawaii (3.71 µg g-1), Texas (4.83 µg g-1), Japan (2.47 µg g-1), and Brazil (1.89 µg g-1). In muscle, the means between Texas (0.75 µg g-1) and Japan (0.75 µg g-1) were the same, and the mean for Brazil (0.13 µg g-1) was the lowest. Among the organs, the highest levels of copper were found in the liver (28.33 µg g-1) followed by the kidney (2.25 µg g-1) and with the lowest levels in the muscle (0.33 µg g-1). This is the first study of copper levels among marine vertebrates in distant parts of the globe using similar comparative filters between different locations. Similar levels in turtles from such distant locations may indicate that there is a pantropical pattern of copper distribution in the biota, and that these animals are subject to the process of bioavailability of this metal in the environment and metabolic regulation.