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.

Organic contamination and multi-biomarker assessment in watersheds of the southern Brazil: an integrated approach using fish from the Astyanax genus.

We aimed to examine the responses of pollution biomarkers in feral fish from Astyanax genus collected at three hydrographic regions in southern Brazil and the capacity of these tools to differentiate between various levels of contamination. To achieve this, levels of organochlorine pesticides (liver), as well as the biomarkers AChE (muscle and brain), TBARS (liver), and EROD (liver) were assessed. Collections were conducted in four municipalities (Alegrete, Caraá, Lavras, and Santa Vitória) during 1 year, encompassing winter and summer. Fish from Alegrete were the most contaminated overall, but animals sampled in Caraá, and Lavras also displayed elevated levels of current-use pesticides. Elevated levels of endosulfans, DDTs, HCHs, and current-use pesticides were accompanied by elevated levels of TBARS in the liver. Conversely, fish from Santa Vitória exhibited the highest levels of PAHs, accompanied by elevated levels of EROD in the liver and reduced levels of AChE in muscle and brain. TBARS proved to be a reliable biomarker for assessing impacts arising from pesticide accumulation, while EROD and AChE served as valuable indicators of impacts resulting from PAHs accumulation. Ultimately, the results obtained in this study demonstrate the reliable use of the proposed biomarkers for tracking biological impacts stemming from aquatic pollution using feral Astyanax as biomonitoring species.

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.

Chlorothalonil as a potential endocrine disruptor in male zebrafish (Danio rerio): Impact on the hypothalamus-pituitary-gonad axis and sperm quality.

Chlorothalonil is a broad-spectrum organochlorine fungicide widely employed in agriculture to control fungal foliar diseases. This fungicide enters aquatic environments through the leaching process, leading to toxicity in non-target organisms. Organic contaminants can impact organism reproduction as they have the potential to interact with the neuroendocrine system. Although there are reports of toxic effects of chlorothalonil, information regarding its impact on reproduction is limited. The aim of the present study was to evaluate the influence of chlorothalonil on male reproductive physiology using the zebrafish (Danio rerio) as ecotoxicological model. Zebrafish were exposed for 7 days to two concentrations of chlorothalonil (0.1 and 10 µg/L) along with a control group (with DMSO - 0.001%). Gene expression of hypothalamus-pituitary-gonad axis components (gnrh2, gnrh3, lhr, fshr, star, hsd17b1, hsd17b3, and cyp19a1), as well as hepatic vitellogenin concentration were assessed. In sperm cells, reactive oxygen species (ROS) content, lipid peroxidation (LPO), mitochondrial functionality, and membrane integrity and fluidity were evaluated. Results indicate that exposure to the higher concentration of chlorothalonil led to a reduction in brain gnr2 expression. In gonads, mRNA levels of lhr, star, and hsd17b1 were decreased at both chlorothalonil concentrations tested. Similarly, hepatic vitellogenin concentration was reduced. Regarding sperm cells, a decreased ROS level was observed, without significant difference in LPO level. Additionally, a higher mitochondrial potential and lower membrane fluidity were observed in zebrafish exposed to chlorothalonil. These findings demonstrate that chlorothalonil acts as an endocrine disruptor, influencing reproductive control mechanisms, as evidenced by changes in expression of genes HPG axis, as well as hepatic vitellogenin concentration. Furthermore, our findings reveal that exposure to this contaminant may compromise the reproductive success of the species, as it affected sperm quality parameters.

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.

Assessment of oxidative stress biomarkers in the threatened annual killifish Austrolebias charrua exposed to Roundup.

This study aimed to analyze the toxic effects of Roundup Transorb® on the endangered Neotropical annual killifish Austrolebias charrua through the assessment of molecular and biochemical biomarkers. The fish were collected in temporary ponds and exposed to environmentally realistic concentrations of the herbicide (5 mg.L-1 for 96 h). The production of ROS, lipid peroxidation, DNA damage, and membrane fluidity were evaluated in the blood cells by flow cytometry. The mRNA expression of the antioxidant-related genes sod2, cat, gstα, atp1a1, gclc, and ucp1 across the brain, liver, and gills was quantified. The acute exposure of annual killifish to Roundup significantly increased ROS production, lipid peroxidation, and DNA damage in their erythrocytes. Likewise, Roundup Transorb® decreased membrane fluidity in the blood cells of the exposed fish. Gene expression analysis revealed that Roundup exposure alters the relative expression of genes associated with oxidative stress and antioxidant defense. Our results give rise to new insights into adaptive mechanisms of A. charrua in response to Roundup. Since Brazilian annual killifishes strongly risk extinction, this study paves the way for developing novel biotechnologies applied to environmental monitoring and aquatic toxicology assessment.

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.

Bioaccumulation and potential impacts of persistent organic pollutants and contaminants of emerging concern in guitarfishes and angelsharks from Southeastern Brazil.

The present study aimed to determine persistent organic pollutants (POPs) and contaminants of emerging concern in endangered angelshark and guitarfish species from southeastern Brazil, and to investigate potential impacts of these compounds on morphometric indexes. Pesticides of emerging concern, pharmaceutical and personal care products (PPCPs), polycyclic aromatic hydrocarbons (PAHs), and polybrominated diphenyl ethers (PBDEs) were determined in hepatic and muscular tissues of Pseudobatos horkelii, P. percellens, Squatina guggenheim, and Zapteryx brevirostris obtained from artisanal and industrial fisheries operating in southeastern Brazil. Accumulation profiles and effects of contaminants on condition factor and hepatosomatic index were investigated. No differences were detected regarding species contaminant concentration, potentially due to similarities in habits, occurrence and trophic position shared by guitarfishes and angelsharks. Polycyclic aromatic hydrocarbons (23.2-495.3 ng g -1), and PPCPs such as diclofenac (

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.

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.

Pantropical distribution of zinc in green turtles (Chelonia mydas): marine vertebrates as sentiel species.

Pollution is one of the biggest threats to marine life and trace elements are among the most toxic pollutants in this environment. Zn is an essential trace element for biota but becomes toxic at high concentrations. Sea turtles are good bioindicators of trace element pollution, due to their longevity and cosmopolitan distribution that allow bioaccumulation for years in their tissues. Determining and comparing Zn concentrations in sea turtles from faraway places is relevant for conservation due to the lack of knowledge of geographically broader distribution patterns of Zn in vertebrates. In this study, comparative analyses of bioaccumulation in the liver, kidney, and muscles of 35 C. mydas from Brazil, Hawaii, the USA (Texas), Japan, and Australia of statistically equal sizes were performed. Zn was found in all specimens, with the highest levels in the liver and kidneys. Specimens from Australia (30.58 µg g-1), Hawaii (31.91 µg g-1), Japan (29.99 µg g-1), and the USA (33.79 µg g-1) showed statistically equal means in the liver. Kidney levels were the same in Japan (35.09 µg g-1) and the USA (37.29 µg g-1) and the same in Australia (23.06 µg g-1) and Hawaii (23.31 µg/g). Specimens from Brazil had the lowest means in both organs (12.17 µg g-1 in the liver and 9.39 µg g-1 in the kidney). The pattern of equal Zn values for most specimens in the liver is an important finding, demonstrating that there are pantropical patterns in the distribution of this metal even in regions so far from each other. A possible explanation is due to the essential nature of this metal linked to metabolic regulation, in addition to the bioavailability for biological absorption in marine environments, such as RS in Brazil, with a lower standard bioavailability also found in other organisms. Therefore, factors such as metabolic regulation and bioavailability indicate that there is a pantropical distribution of Zn in marine biota and green turtles can be a useful model as a sentinel species.

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.

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.

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.

Ecotoxicological impacts of the Fundão dam failure in freshwater fish community: Metal bioaccumulation, biochemical, genetic and histopathological effects.

This study investigates the ecotoxicological impacts of the Fundão dam rupture, one of the major environmental disaster that occurred in Brazil and in the world mining industry history, through multi-biomarkers responses and metals bioaccumulation in the fish community of different trophic levels. Specimens of the fishes (omnivorous/herbivorous and carnivorous) were collected along the Doce River channel and its affluent Guandú River, and in different lakes and coastal lagoons adjacent to the river channel, in the Espirito Santo State, Southeast of Brazil. Four sampling collections were carried out over two years (2018 to 2020, during dry and rainy seasons). For both trophic groups the biomarkers responses indicated physiological alterations related to metals exposure and showed strong seasonal variations. The principal component analysis and integrated biomarker response index showed that DNA damage and lipid peroxidation were more associated with dry season 2 (Sep/Oct 2019) and the oxidative damage in proteins, metallothioneins concentration and the activity of superoxide dismutase in the gills showed a greater association with rainy season 2 (Jan/Feb 2020). On the other hand, the enzymes of energy metabolism, catalase and histological damage in the liver and the gills, were more associated with the dry and rainy campaigns of the first year of monitoring. The multivariate approach also suggested a temporal intensification in the bioaccumulation of metals and biological effects in the lacustrine environments. Thus, these results demonstrate that the release of mineral residues from the rupture of the Fundão mine dam affects the health status of the fish from the Doce River basin, provoking metals bioaccumulation, hepatic and branchial damage in the fish besides inducing of enzyme activity related to metal contamination, even four years after the rupture.

Marine shrimps as biomonitors of the Fundão (Brazil) mine dam disaster: A multi-biomarker approach.

The disruption of the Fundão dam released 43 million m3 of mine tailings into the Doce River until it flowed into the ocean through the estuary. The mine tailing changed the composition of metals in water and sediment, creating a challenging scenario for the local biota. We used multivariate analyzes and the integrated biomarker response index (IBR) to assess the impact of mine tailings on the bioaccumulation profile (As, Cd, Cr, Cu, Fe, Mn, Pb and Zn) as well as the biomarkers response in gills, hepatopancreas and muscle of shrimps sampled from different sectors during two dry seasons (dry1 and dry2) (Sep/Oct 2018; 2019) and two wet seasons (wet1 and wet2) (Jan/feb 2019; 2020). There was seasonal and local effect under bioaccumulation and biomarker response revealing that the pattern responses seen in each sector sampled changed according to the season. The greater IBR added to the strong association among the most metals tissue content (Cd, Cr, Cu and Mn) and sectors sampled during dry 1 suggests greater bioavailability of these metals to the environment in this period. Estuarine sectors stand out for high Fe bioavailability, especially during wet1, which seems to be associated with greater metallothionein content in hepatopancreas of shrimps. Native species of marine shrimps proved to be successful indicators of sediment quality besides being sensitive to water contamination by metals. The multi-biomarkers approach added to multivariate analysis supports the temporal and seasonal effects, signalizing the importance of continuous monitoring of the estuarine region to better know about the bioavailability of these metals, mainly Fe, and their long-term effects on the local biota.

Impacts of tailings of Fundão dam (Brazil) rupture on marine fish: Metals bioaccumulation and physiological responses.

This study evaluated the impacts of the mining tailings after the rupture of the Fundão dam on fish communities on the Atlantic Ocean southeast coast. Four sample collections were carried out over two years (2018-2020), in seasonal periods. Omnivorous/herbivorous and carnivorous fish were collected for analysis of metal bioaccumulation, multibiomarkers of environmental contamination and histopathology. Metal bioaccumulation was stronger correlated in carnivorous fish in the dry-2018 collection, besides higher activity of antioxidant enzymes, energy metabolism and higher morphological damage; however, there was less oxidative damage and less metallothioneins induction, and these variables were strongly associated with the wet-2020 collection. In a temporal view, it was possible to observe a reduction in metal levels in fish, except in the mouth of the Doce River. These events can be explained by seasonal natural events, which tend the resuspension and boost metal levels, mainly in the mouth region during the rainy season.

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.

Assessing multigenerational exposure to metals in elasmobranchs: Maternal transfer of contaminants in a yolk-sac viviparous species.

This study aimed to assess the maternal offloading of metals in the Brazilian guitarfishPseudobatos horkelii through determining essential (Cr, Cu, Fe) and non-essential (Cd, Hg, Pb) metal concentrations along two generations of this species: pregnant females and offspring. The maximum transfer capacity (ECER) and offspring/mother ratio were calculated to estimate the extent of offloading, as well as the proportion of contaminants presented in offspring related to the maternal concentrations. Transfer efficiency was element-dependent. Chromium had the highest ECER (99.7%), followed by Hg (67.7%). Other essential metals were less transferred (9.6-35.6%) and Cd and Pb were not detected in uterine content samples. The relationships between maternal length and concentration, as well as transfer capacity were not significant, indicating that females might be continuously exposed, and that offloading might not excrete metals efficiently. On the other hand, embryos are exposed to these elements which could impair embryonic development.

Metal accumulation induces oxidative stress and alters carbonic anhydrase activity in corals and symbionts from the largest reef complex in the South Atlantic ocean.

We evaluated the influence of metal accumulation on the oxidative status [lipid peroxidation (LPO) and total antioxidant capacity (TAC)] and carbonic anhydrase (CA) activity in host and symbionts of the coral Mussismilia harttii and the hydrocoral Millepora alcicornis collected in Abrolhos Reef Banks (Northeast Brazil), potentially impacted by a major mine dam rupture. Considering metal levels measured in reefs worldwide, Abrolhos corals had higher Fe and Mn levels than expected for preserved offshore reefs. Increasing concentrations of arsenic (As), chromium (Cr) and manganese (Mn) drove inhibition of CA and increased oxidative damage in the hydrocoral M. alcicornis. The impairment of enzymatic activity in the symbiotic algae of M. alcicornis may be related to the oxidative stress condition. The hydrocoral M. alcicornis was more affected by metals than the coral M. harttii, which did not show the expected CA inhibition after metal exposure. Our results suggest that CA activity can be applied as a complementary biomarker to evaluate the physiological impacts of environmental metal contamination in reefs. Also, the metal levels and biochemical biomarkers reported in the present study may provide reference data to monitor the health of reefs impacted by a relevant dam rupture.