Behavioral and electrophysiological study in Colossoma macropomum treated with different concentrations of Nepeta cataria oil in an immersion bath revealed a therapeutic window for anesthesia.

The purpose of this study was to characterize the activity of essential oils from Nepeta Cataria (EON) at concentrations of 125 µ L  L-1, 150 µ L  L-1, 175 µ L  L-1, and 200 µ L  L-1 on the behavior of loss of the posture reflex and recovery of the posture reflex and electrocardiographic activity and recording of the opercular beat of Colossoma macropomum during immersion bathing for a period of 5 min, in order to obtain a window for safe use during anesthesia. The fish (23.38 ± 3.5 g) were assigned to the following experiments: experiment 1 (latency to loss and recovery of the posture reflex): (a) 125 µ L  L-1, (b) 150 µ L  L-1, (c) 175 µ L  L-1, and (d) 200 µ L  L-1 (n = 9) per group. Experiment 2 (electrocardiographic and heartbeat recordings): (a) control group; (b) vehicle control group (2 ml of alcohol per liter of water), (c) 125 µ L  L-1, (d) 150 µ L  L-1, (e) 175 µ L  L-1, and (f) 200 µ L  L-1 (n = 9), per group. All the concentrations used showed efficacy in inducing loss of the posture reflex and reversibility with recovery of the posture reflex, but the electrocardiographic recordings indicated morphographic changes such as bradycardia during induction and p wave apiculation during recovery at the highest concentrations tested. In this way, we suggest a safe use window for short-term anesthesia with EON in the concentration range of 125 to 150 µ L  L-1 for juvenile Colossoma macropomum.

Multiple anthropogenic influences in the Pará River (Amazonia, Brazil): A spatial-temporal ecotoxicological monitoring in abiotic and biotic compartments.

The mineral wealth of the Amazon region is prone to intense exploration with consequent metals mobilization in ecosystems. Besides that, a number of other anthropogenic activities contribute to the imbalance of this important environment. The Pará River is an important water body in the Amazon basin and is under multiple anthropogenic influences, including disorderly urbanization, port activities and processing of minerals such as bauxite and kaolin. In this study, metals concentrations (Al, Cr, Pb, Ba, Ni, and Mn) in water, sediment and organisms (the fish Cheirocerus goeldii and the shrimp Macrobrachium amazonicum) and biochemical biomarkers (total antioxidant capacity, ACAP and lipid peroxidation, LPO) were analyzed along five points in the Para River with different distances to a center of anthropogenic activity, in three seasonal periods (rainy, transition and dry). Metals concentrations were similar among sites in each seasonal period but Aluminium (Al) presented the highest concentrations among all analyzed metals both in sediment and water considering all sampling points and all seasonal periods. In the dissolved fraction, Al had values above those established by the local environmental agency, especially during the rainy season. In the biotic compartment, both fish and shrimps showed higher concentrations of metals (Al and Ba) in the rainy season compared to the other seasons. Shrimp was more responsive to metal accumulation than fish, showing an adaptive response of biomarkers. Fish showed an increase of LPO in gills for individuals from the point of greater anthropogenic interference during the rainy season, but no differences in metal accumulation. We conclude that there is a seasonal pattern of metals concentration in different environmental compartments. Metal concentration in organisms and biomarkers responses, showed the effect of anthropogenic influences, which was not evident in results from chemical analyses alone, due to the intensive hydrodynamics in the region.

Distribution of metals in different environmental compartments and oxidative stress biomarkers in Bryconops caudomaculatus (Osteichthyes: Characiformes) from a bauxite mining area in the Eastern Amazon.

The Eastern Amazon is rich in bauxite ore. The extraction and processing of bauxite lead to the mobilization of Aluminum (Al) and other metals in environmental. We evaluated the metals (Al, Mn, Ba, and Cr) concentration in tissue, water, and sediment associated with antioxidant and oxidative damage responses in Bryconops caudomaculatus. The samplings were done in two hydrological periods (post-rain and post-dry periods) and at three points, located at two rivers: one in the surroundings of the mining area (P1) and other inside the mining area, upstream (P2), and downstream (P3). Defense antioxidant system biomarkers analyzed were total antioxidant capacity (ACAP) and glutathione-S-transferase (GST) activity. As an oxidative damage biomarker, the lipoperoxidation (LPO) was evaluated. Metals concentrations in the water and sediment were higher in the post-rain period compared to post-dry period. The water samples were acidic, with dissolved Al concentrations above the values established by local legislation at all points. In the gills, the metals accumulation was higher in fish from in the surrounding and upstream sites, and in the liver, was higher in fish from downstream site. Fish from the surrounding had increased antioxidant defenses, with higher ACAP in all tissues and higher GST in the gills. Consequently, they had lower levels of LPO. Fish from the mining area had decreased antioxidant defenses, with lower ACAP in all tissues and lower GST in the gills. Consequently, they had higher levels of LPO, indicating oxidative stress. The fish muscle was not responsive to GST and LPO at all sites. We conclude that the oxidative stress observed in the gills and liver of B. caudomaculatus from the area modified by the mining activity reflected the local anthropogenic impact status.

Cardiac responses in Crassostrea gasar: An experimental approach of how the tidal cycle influences the heart function of the mangrove oyster.

Cardiac physiological studies in oysters are scarce and these kinds of responses can be key issues for understanding behavioral and baseline adaptive responses. In this study we characterized the electrocardiogram (ECG) tracing patterns, wave intervals (RR; QT), and complex duration (QRS) of Crassostrea gasar during immersion followed by air exposure, simulating what occurs in a tide cycle. Initially, the ECG was analyzed in the oysters under immersion for 30 min to evaluate the basal recordings (immersion exposure condition). Then, the same animals were removed from the water and the ECG was analyzed for another 30 min to assess cardiac activity under air exposure (air exposure condition). For this, a technique of cardiac recordings was developed, allowing us to analyze, beyond ECG, other cardiac parameters such as the heart rate (HR) in beats per minute, the spectral power of HR, and the amplitude. The basal cardiac parameters analyzed in oysters in the immerse condition clearly show the waves and intervals (R-R: 11.03 ± 0.63 s, P-Q: 2.09 ± 0.06 s), with a normal and regular electrocardiographic tracing and sinus rhythm without alterations. When in the air exposure condition, oysters maintained the ECG tracing of sinus rhythm, but with changes in intervals and a prolonged isoelectric period. Moreover, in this condition, oysters presented a biphasic response: initially (phase I), heart rate increased (9.83 ± 0.98 BPM), and consequently the R-R and P-Q intervals decreased (5.86 ± 2.01 s e 1.91 ± 0.13 s, respectively); in phase II, heart rate (3.68 ± 0.82 BPM) and spectral power (21.26 ± 5.44 mV2/Hz x 10-3) decreased and consequently, the R-R interval increased (14.83 ± 2.92 s). But, the P-Q interval remained (2.45 ± 0.65 s) in phase II. The QRS complex of oysters in both phases decreased (Phase I: 0.57 ± 0.13 s; Phase II: 0.62 ± 0.05 s) compared to the immersion exposure condition (0.79 ± 0.09 s). We conclude that air exposure affected cardiac function in C. gasar leading to arrhythmia in response to the beginning of air exposure, as a means of maintaining oxygen supply, followed by bradycardia to decrease metabolism as a survival strategy. The basal responses of the mangrove oyster in the physiological modulation against the environmental factors of the tidal regime provide information about the species for possible application as model organisms in studies of toxicological evaluation of chemical products and in conservation and sustainability studies.

Evaluation of cardiotoxicity in Amazonian fish Bryconops caudomaculatus by acute exposure to aluminium in an acidic environment.

Aluminium (Al) is soluble in acidic waters and may become toxic to organisms. In this study, the acute effects of two Al concentrations were evaluated in the Amazonian fish Bryconops caudomaculatus. Antioxidant responses and lipid damage were assessed in gills, liver and muscle, along with the electrocardiography (ECG) and characterization of cardiac complex and wave intervals. Fish were essayed as follows: two control groups at neutral and acidic pH and two exposure groups at acidic pH (0.3 mg/L and 3.0 mg/L Al). Water samples were collected at 0h, 24h and 48h, for chloride (Cl-), fluoride (F-) and sulphate (SO42-) ion analyses, while total Al was quantified in muscle. Concentrations of Cl- and SO42- were constant over time whereas F- was not detected. Total Al concentrations in water and muscle were concentration-dependent. Antioxidant responses, total antioxidant capacity against peroxyl radicals (ACAP) and glutathione S-transferase were not triggered in fish tissues exposed to 0.3 mg/L Al; however, fish exposed to 3.0 mg/L Al presented increased and reduced ACAP in gills and liver, respectively. No changes in lipoperoxidation levels occurred among groups. Fish exposed to 0.3 mg/L Al showed prolonged intervals in ECG as a reflection of low heart rate (HR), with sinus bradycardia. Moreover, there was a marked prolongation of the PQ interval (time between the atrial activity and the start of ventricular activity), indicating interference on the cardiac cell automaticity. Fish exposed to the highest concentration of Al showed reduced wave intervals as a consequence of increased HR, with sinus arrhythmia, while ECG tracings did not present P waves (atrial contraction), indicating an atrioventricular blockade. In conclusion, 48h exposure sufficed to cause cardiotoxicity in B. caudomaculatus at either Al concentration. However, as oxidative stress was not observed, such cardiac alterations seem to be reversible under the experimental conditions established herein.

The legacy of artisanal gold mining and its impact on fish health from Tapajós Amazonian region: A multi-biomarker approach.

Tapajós Region, is an area with intense historical artisanal and small-scale gold mining. Therefore, the core objective of this study was to evaluate the environmental status of different rivers located in this region, using biomarker endpoints in Serrasalmus rhombeus as a tool. Fish and sediment were collected from two rivers, Tropas and Crepori, affluent of Tapajós River, located inside a Federal Protection Area and in a Reference site. Mercury concentration in sediment and fish were traced, and biomarkers in gills and liver were analyzed. Results showed a clear difference between these two rivers compared to the Reference site. Fish tissues presented biomarker responses according to the site of collection. Catalase (CAT) activity was statistically higher in fish gills from Crepori, confirming the capacity of mercury interference with redox equilibrium. High levels of lipid peroxidation were also noted to contribute greatly in incidence of morphological changes in the liver and gills, suggesting that mercury bioaccumulation during continuous exposure promote biological responses in a cumulative manner, from molecules to tissues. This study also indicates adaptation in fish defense mechanisms given the conditions in the Tropas River, as well as a variation in biomarker responses to that of the Crepori river. In summary, Tapajós affluents presented high mercury levels in fish tissues leading to biomarker responses, demonstrating a hazardous signal of a long history of mercury pollution.

Seasonal modulation of oxidative stress biomarkers in mangrove oyster (Crassostrea gasar) from an Amazon estuary.

Estuaries are the final destination of many pollutants derived from anthropogenic activity. Therefore, it is difficult to find this kind of ecosystem in a pristine condition. In this context, biomonitoring studies that characterize the organism's conditions against the environment' s natural variation are essential for future impact analysis due to anthropic activity. The present study aims to characterize the natural modulation of biochemical biomarkers in oysters Crassostrea gasar. The research was conducted in Japerica Bay, an estuary region located in the Eastern Amazon (Pará, Brazil), which has remained in pristine condition for many years. The samplings were carried out throughout one year during the rainy-dry transition period (June/2013), dry period (September/2013), dry-rainy transition period (November / 2013), and rainy period (February / 2014) in the lower and upper estuary. The activity of glutathione-S-transferase (GST) and total antioxidant capacity (ACAP) were evaluated as biomarkers of exposure and lipid peroxidation (LPO) as an effect biomarker. In gills, GST decreased during the rainy season in both sites and increased during the salinity peak (dry-rainy transition period) for the upper estuary's organisms. In this organ, the lowest levels of LPO occurred during the dry season for both points. There was an induction of ACAP in muscle during the rainy-dry transition period compared to the dry and dry-rainy transition periods for the lower estuary's organisms, and there were no differences for GST suggesting low tissue sensitivity. There was an increase in LPO during the rainy season compared to the rainy-dry transition period for the lower estuaries animals. Biomarkers in gills suggest a metabolic challenge to the rainy season and stability during the dry season. The species shows high viability of use in biomonitoring programs. However, these seasonality-induced alterations in biomarkers responses must be taken into account to interpret the results.

Spatial distribution of butyltins and imposex in eastern Brazilian Amazon.

Spatial distribution of Butyltins (BTs) in surface sediments and gastropod tissues were quantified, with samples taken from an estuarine system of the Eastern Amazon, Brazil. The imposex incidence was also assessed using Thaisella coronata (Gastropoda, Mollusca). The sediment sampling was carried out at 19 sites and T. coronata in 6 of those. The highest BTs levels were detected in sediments of a Marine Extractive Reserve (27.1 ng Sn g-1) and in an urban area (19.8 ng Sn g-1). In T. coronata tissues, BTs levels ranged from <5 to 142 ng Sn g-1. Imposex incidence ranged from 0% to 100% and VDSII (penile papillae) was registered in gastropods from 5 out of 6 sampled sites. The results suggest that BTs concentrations are related to sampled area use, but also to the local hydrodynamics, highlighting the importance of an effective control in the use of TBT-based antifouling paints in the Eastern Amazon.

Eugenol induces body immobilization yet evoking an increased neuronal excitability in fish during short-term baths.

Several studies have suggested eugenol as a suitable anaesthetic for fish. However, it has also been regarded as a toxic and aversive substance to several aquatic organisms, including fish. This study sought to assess the eugenol-induced behavioural alterations and its seizurogenic potential to fish. Moreover, a distinctive methodology for an in vivo evaluation of the brain activity was also presented. Prior to the evaluation of eugenol-induced responses, fish were exposed to pentylenetetrazole (PTZ), to characterize any seizure-like patterns. Antagonizing responses to PTZ were assessed in fish receiving diazepam (BDZ) and subsequently exposed to PTZ. Tambaqui fish juveniles, Colossoma macropomum (15.8 ± 2.8 g) were used as models and assayed as follows: (i) fish exposed to PTZ (15 mM) and (ii) fish receiving a dose of BDZ (10 mg Kg-1) and later exposed to PTZ (15 mM) (BDZ-PTZ group). Thereafter, fish were evaluated throughout (iii) eugenol exposure at 65 µL L-1 (ethanolic solution) and recovery. Control fish and a vehicle control group (ethanol at 585 µL L-1) were also established. PTZ baths elicited body immobilization preceded by hyperactivity in a stereotyped seizure-like behaviour with increased EEG wave amplitude and frequency. PTZ effects in the brain were attenuated by a pre-administration of BDZ. Upon eugenol exposure, tambaqui had an intense neuronal excitability, showing a clonus-like seizure behaviour, also corroborated by the EEG patterns, which were consistent with a seizure-like response. Responses of eugenol-exposed fish resembled those of the PZT-exposed animals, with epileptiform discharges. EMG was in line with the EEG modulation, showing increased tracing oscillations and higher mean amplitudes in PTZ-exposed fish whereas in BDZ-PTZ group muscle contraction was less frequent and powerful. Fish exposed to eugenol showed initially some muscle activity followed by a loss of muscle tonus over time. In summary, our results showed that upon eugenol exposure, although a time-dependent body immobilization was attained, fish presented an intense neuronal excitability comparable to that evoked by PTZ. Eugenol failed to promote depression of the CNS and therefore may be not suitable to be used for general anaesthesia of C. macropomum. As eugenol could be implicated in seizurogenesis and be potentially toxic to the fish brain, protocols suggesting the broad use of eugenol for short-term anaesthesia or euthanasia of fish should be carefully revised, as it raises important concerns in terms of ethics and fish welfare.

Evaluation of metal contamination effects in piranhas through biomonitoring and multi biomarkers approach.

The present field study aimed to assess the water quality of the Itacaiúnas River, located at the eastern part of the Brazilian Amazon, using water's physicochemical parameters, ecological risk assessment in sediments, biomarkers and metal bioaccumulation in piranhas at two points: upstream (P1) and downstream (P2), and the type of season (Dry and Rainy). We revealed a significant difference between the points and the seasons. Concerning, the concentration of metals (Cd, Cu, Zn, Cr, and Hg) in sediment and water, we reported significant concentrations of Cd and Cu especially on P2 at dry season. The fish gills collected in P2 showed various types of damages (moderate and severe), furthermore, the Degree of Tissue Change (DTC) reported a significant difference between points, highlighting the dreadful condition in animals' health originated from this point. In terms of the biotransformation enzyme, the GST activity was higher in fishes from P2 in both seasons. The obtained results showed clear signs of stress in fish from the downstream point. Linear correlation analysis exhibited that the biomarkers' response could be linked to the detected metals bioaccumulation. This field investigation provides baseline data on pollution status in this region and the results showed that although the overall potential ecological risks of the metals were considered low at our sampling sites including cadmium, however, Cd posed a noteworthy monomial potential ecological risk factor. Strong evidence of correlation was obtained between Cd in the environment with the gills' damage in fishes from P2. The results also indicated that S. rhombeus could be useful for biomonitoring species for assessing metal contamination.

Electrophysiological responses in Amazonian fish species Bryconops caudomaculatus (Osteichthyes: Characiformes) as biomarkers of xenobiotic toxicity.

Sublethal exposures to environmental pollutants may cause changes in physiological parameters. Thus, knowledge of basal physiological rates of the species and the development of methods to quantify these rates are extremely important. Considering the scarcity of cardiac and muscle physiological studies in native Amazonian fish species and that no evaluation of electrophysiological responses by exposure to a stressor has been reported in Bryconops caudomaculatus, the aim of this study was to develop techniques of electromyographic and electrocardiographic recordings of normal responses, during toxicity induction and short-term recovery. A total of 9 animals were used, divided into two groups: control group (n = 4) and treated group (n = 5), with records lasting 5 min. The results showed that the basal electromyographic records indicate that the studied species has a very intense swimming activity, whereas the basal cardiac parameters clearly showed the patterns in P wave tracing, QRS complex, T wave and Q-T and R-R intervals. During exposure to the stressor, muscle activity ceased presenting intense decrease and myorelaxant effect expected. Electrocardiographic responses confirmed cardiotoxicity with intense bradycardia, ventricular bigeminism, prolongation of QRS complex duration and cardiac arrhythmias, indicating cardiac dysfunction. It was concluded that the electrophysiological responses are excellent biomarkers and showed the susceptibility of the species to the tested substance. In addition, the electrocardiogram and the electromyogram are excellent techniques to reflect the degree of environmental stress when organisms are exposed to toxic substances in the environment.

Methylmercury intoxication and cortical ischemia: Pre-clinical study of their comorbidity.

Stroke is one of the main causes of human disability worldwide. Ischemic stroke is mostly characterized by metabolic collapse and fast tissue damage, followed by secondary damage in adjacent regions not previously affected. Heavy metals intoxication can be associated with stroke incidence, because of their damaging action in the vascular system. Mercury, in particular, possesses a high tropism by metabolically active regions, such as the brain. In the present study we sought to evaluate whether methylmercury (MeHg) intoxication can aggravate the tissue damage caused by an ischemic stroke induced by microinjections of endothelin-1 (ET-1) into the motor cortex of adult rats. Following MeHg intoxication by gavage (0.04 mg/kg/day) during 60 days, the animals were injected with ET-1 (1 µl, 40 pmol/µl) or vehicle (1 µl). After 7 days, all animals were submitted to behavioral tests and then their brains were processed to biochemical and immunohistochemical analyses. We observed that long-term MeHg intoxication promoted a significant Hg deposits in the motor cortex, with concomitant increase of microglial response, followed by reduction of the neuronal population following ischemia and MeHg intoxication, as well as disturbance in the antioxidant defense mechanisms by misbalance of oxidative biochemistry with increase of both lipid peroxidation and nitrite levels, associated to behavioral deficits. MeHg exposure and cortical ischemia demonstrated that both injuries are able of causing significant neurobehavioural impairments in motor coordination and learning accompanied of an exacerbated microglial activation, oxidative stress and neuronal loss in the motor cortex, indicating that MeHg as a source of metabolic disturbance can act as an important increasing factor of ischemic events in the brain.

Influence of seasonality on the natural modulation of oxidative stress biomarkers in mangrove crab Ucides cordatus (Brachyura, Ucididae).

The estuaries are characterized by the formation of salinity gradients and are frequently associated with anthropic activities. Studies that show the condition of organisms against the natural environmental variations are fundamental for future evaluation of environmental change due to anthropic impacts. Biomarkers are useful tools defined as biological changes in low levels of organization induced by stressors and considered as predictive responses of more severe effects. The aim of this study was to characterize the modulation of biochemical biomarkers in Ucides cordatus against the natural variation a pristine mangrove forest in an Amazonic estuary (Japerica Bay). The samplings were carried out throughout one year at rainy-dry transitional period (June/2013), dry period (September/ 2013), dry-rainy transitional period (November/ 2013) and rainy period (February/ 2014) in the lower, middle and upper estuary. The biochemical biomarkers evaluated were glutathione-S-transferase (GST), glutamate cysteine-ligase (GCL), total antioxidant capacity (ACAP), reduced glutathione (GSH) concentration and the lipid peroxidation (LPO). There was a decrease in the activity of both enzymes, GSH concentration and lower LPO in the rainy-dry transitional period for the organisms of all sites, pointing to this period as a season of greater metabolic stability. The organisms, especially those of the middle estuary, presented physiological plasticity against the variations of the environment which they are submitted. Seasonality, more than salinity influence due to position in estuary, is the main modulator of biomarkers. Also, our results indicate that the species presents high viability for biomonitoring studies.

Low doses of methylmercury exposure during adulthood in rats display oxidative stress, neurodegeneration in the motor cortex and lead to impairment of motor skills.

Despite the vast distribution among tissues, the central nervous system (CNS) represents the main target of methylmercury (MeHg) toxicity. However, few studies have evaluated the effects of MeHg exposure on the CNS at equivalent doses to human environmental exposure. In our study, we evaluated the motor cortex, an important area of motor control, in adult rats chronically exposed to MeHg in a concentration equivalent to those found in fish-eating populations exposed to mercury (Hg). The parameters evaluated were total Hg accumulation, oxidative stress, tissue damage, and behavioral assessment in functional actions that involved this cortical region. Our results show in exposed animals a significantly greater level of Hg in the motor cortex; increase of nitrite levels and lipid peroxidation, associated with decreased antioxidant capacity against peroxyl radicals; reduction of neuronal and astrocyte density; and poor coordination and motor learning impairment. Our data showed that chronic exposure at low doses to MeHg is capable of promoting damages to the motor cortex of adult animals, with changes in oxidative biochemistry misbalance, neurodegeneration, and motor function impairment.

Proteomic approach underlying the hippocampal neurodegeneration caused by low doses of methylmercury after long-term exposure in adult rats.

Methylmercury (MeHg) is an important toxicant that causes cognitive dysfunctions in humans. This study aimed to investigate the proteomic and biochemical alterations of the hippocampus associated with behavioural consequences of low doses of MeHg in a long-term exposure model, and to realistically mimic in vivo the result of human exposure to this toxicant. Adult Wistar male rats were exposed to a dose of MeHg at 0.04 mg kg-1 day-1 by gavage for 60 days. Total mercury (Hg) content was significantly increased in the hippocampal parenchyma. The increase in the Hg levels was capable of reducing neuron and astrocyte cell density in the CA1, CA3, hilus and dentate gyrus regions, increasing both malondialdehyde and nitrite levels and decreasing antioxidant capacity against peroxyl radicals. The proteomic analysis detected 1041 proteins with altered expression due to MeHg exposure, including 364 proteins with no expression, 295 proteins with de novo expression and 382 proteins with up- or down-regulated expression. This proteomic approach revealed alterations in pathways related to chemical synapses, metabolism, amino acid transport, cell energy, neurodegenerative processes and myelin maintenance. Therefore, even at low doses of MeHg exposure, it is possible to cause hippocampal damage in adult rats at many organisational levels, triggering oxidative stress and proteome misbalance, featuring a neurodegenerative process and culminating in long- and short-term memory and learning deficits.

Hippocampal Dysfunction Provoked by Mercury Chloride Exposure: Evaluation of Cognitive Impairment, Oxidative Stress, Tissue Injury and Nature of Cell Death.

Mercury (Hg) is a highly toxic metal, which can be found in its inorganic form in the environment. This form presents lower liposolubility and lower absorption in the body. In order to elucidate the possible toxicity of inorganic Hg in the hippocampus, we investigated the potential of low doses of mercury chloride (HgCl2) to promote hippocampal dysfunction by employing a chronic exposure model. For this, 56 rats were exposed to HgCl2 (0.375 mg/kg/day) via the oral route for 45 days. After the exposure period, the animals were submitted to the cognitive test of fear memory. The hippocampus was collected for the measurement of total Hg levels, analysis of oxidative stress, and evaluation of cytotoxicity, apoptosis, and tissue injury. It was observed that chronic exposure to inorganic Hg promotes an increase in mercury levels in this region and damage to short- and long-term memory. Furthermore, we found that this exposure model provoked oxidative stress, which led to cytotoxicity and cell death by apoptosis, affecting astrocytes and neurons in the hippocampus. Our study demonstrated that inorganic Hg, even with its low liposolubility, is able to produce deleterious effects in the central nervous system, resulting in cognitive impairment and hippocampal damage when administered for a long time at low doses in rats.

Metal and metalloid distribution in different environmental compartments of the middle Xingu River in the Amazon, Brazil.

Concentrations of As, Cd, Cu, Cr, Pb, Hg, and Ni were analyzed during rainy and dry seasons in water, sediment, soil, and two fish species. The analysis took place at four points in the Xingu River, one point in the Fresco River, and two mining pits in the southeastern area of the Eastern Amazon, Brazil. In the water, the total concentration of As (>0.14µg/L) was higher than the local reference values at all sampling points and in both seasons. Ordination analysis (PCA) highlighted As and Cu elements in the water. PERMANOVA showed that the metals behaved differently in the water throughout the monitored season and between sampling points. The sites with mining activity were the regions that were the most contaminated by metals. Samples of sediment (Ni>18mg/kg and Cr>37.30mg/kg) and soil (Pb>72mg/kg, Cr>75mg/kg and Ni>30mg/Kg) showed concentrations above the recommended by local legislation. Metal values in the muscle of both fish species were relatively low at all sampling points and in both monitored seasons. Concentrations in water, sediment, and soil showed that some points of the Xingu River, Fresco River and mining pits are contaminated by trace elements, mainly As, Hg, Cr, Pb, and Ni. This was the first study about trace elements in the Middle Xingu River, which leads us to conclude that rainfall and cassiterite mining activities strongly influence the mobilization of metals, especially in abiotic compartments. However, the fish analyzed did not exhibit relevant levels of contamination. This indicates low risk for human consumption. Additionally, results highlight the need to establish local criteria to define contamination limits for different metals while taking into account local geochemistry particularities and biome diversity.

Evaluation of the use of Olivella minuta (Gastropoda, Olividae) and Hastula cinerea (Gastropoda, Terebridae) as TBT sentinels for sandy coastal habitats.

Tributyltin (TBT) contamination is still recorded in the environment even after its ban in antifouling paints. Since most biomonitors of TBT contamination, through imposex evaluation, are hard-bottom gastropods, the identification of soft-bottom sentinels has become useful for regions where rocky shores and coral reefs are absent. Thus, an evaluation of Olivella minuta and Hastula cinerea as monitors of TBT contamination was performed in two sandy beaches located under influence area of São Sebastião harbor (São Paulo state, Brazil), where previous and simultaneous studies have reported environmental contamination by TBT. In addition, the imposex occurrence in H. cinerea was assessed in an area with low marine traffic (Una beach), also located in São Paulo State. A moderate imposex incidence in O. minuta was detected in Pernambuco (% I = 9.36, RPLI = 4.49 and RPLIstand = 4.27) and Barequeçaba (% I = 2.42, RPLI = 0.36 and RPLIstand = 0.81) beaches, indicating TBT contamination. In contrast, more severe levels of imposex were recorded for H. cinerea in Una beach (% I = 12.45) and mainly in Barequeçaba beach (% I = 98.92, RPLI = 26.65). Our results suggest that O. minuta and H. cinerea have good potential as biomonitors for TBT based on their wide geographical distribution, common occurrence in different coastal sediment habitats, easy collection, and association with TBT-contaminated sediments.

Influence of a toxic Microcystis aeruginosa strain on glutathione synthesis and glutathione-S-transferase activity in common carp Cyprinus carpio (Teleostei: Cyprinidae).

We evaluated the effects of aqueous extracts of the cyanobacterium-producing microcystin (MC), Microcystis aeruginosa (strain RST9501), on detoxification capacity and glutathione (GSH) synthesis in liver, brain, gill, and muscle-as well as apoptotic protease (calpain) activity in liver and brain-in the common carp Cyprinus carpio (Teleostei: Cyprinidae). Experimental groups were defined as follows: (1) control (CTR); (2) carp treated with an aqueous extract from the toxic cyanobacteria M. aeruginosa in a final MC concentration of 25 µg/kg (MC 25); and (3) carp treated with an aqueous extract from the toxic cyanobacteria M. aeruginosa in a final MC concentration of 50 µg/kg (MC 50). Carp were gavaged with a cyanobacterial aqueous solution or MilliQ water (CTR group). The experiment was conducted for period of 48 h comprising two gavages with a 24-h interval between them. Some of the parameters analyzed in liver, brain, gill, and muscle included activity of the enzymes glutathione-S-transferase (GST), glutamate cysteine ligase (GCL), glutathione reductase (GR), and GSH concentration. We also evaluated GST pi concentration by Western blot as well as calpain activity in liver and brain samples. In carp liver from the MC 50 group, we observed a decrease in GST and GCL activity, which was accompanied by a decreased GSH concentration. In addition, liver calpain activity was highly induced in carp at both MC doses. Thus, MC ingestion affected the liver antioxidant status through decreasing the GSH concentration and the activity of the enzyme involved in its synthesis (GCL). It also decreased the MC detoxification capacity of the liver because total GST activity decreased, a result that cannot be ascribed to GST pi levels. Because GSH acts as an uncompetitive inhibitor of calpain, its decrease should explain the higher activity of this apoptotic enzyme. The main goal of this study was to show that a decrease in GSH concentration is related to decreased activity of GCL, the limiting enzyme involved in GSH synthesis. Because MCs are phosphatase inhibitors and GCL is allosterically inhibited by phosphorylation, the cellular hyperphosphorylation state induced by MC exposure could act as a modulator factor for antioxidant defenses.

A method to measure total antioxidant capacity against peroxyl radicals in aquatic organisms: application to evaluate microcystins toxicity.

Determination of total antioxidant capacity, instead of the measurements of limited number of antioxidants, is very important for the understanding of how antioxidants interact with reactive oxygen species (ROS). Several techniques already exist with this propose, although some of them are extremely time-consuming. A new methodology is proposed, based on the detection of ROS by fluorometry (ex/em: 485/520 nm) employing 2',7'-dichlorofluorescein diacetate (H(2)DCF-DA) as substrate. Supernatant of homogenized samples from different organs (gill, muscle, liver, and brain) of the teleost fish Jenynsia multidentata (Anaplebidae) were exposed to peroxyl radicals generated by thermal (35 degrees C) decomposition of 2,2'-azobis (2 methylpropionamidine) dihydrochloride (ABAP, 4 mM). Different protein concentrations (0.5, 1, 2 and 8 mg/ml) were assayed to get the best signal and curve fitting of fluorescence data over time (30 min). Total antioxidant capacity against peroxyl radicals was estimated as the difference in ROS area with and without ABAP, relative to the fluorescence registered without ABAP. For application of this methodology, J. multidentata specimens were exposed for 24 h to microcystins, cyanotoxins known to induce oxidative stress. Almost all organs showed a lower antioxidant capacity (p<0.05) in samples with 8 mg proteins/ml, when compared to protein content of 1-2 mg/ml. In liver samples, higher (p<0.05) free iron content was determined in samples with 8 mg proteins/ml. Sensitivity test employing GSH spiked in homogenized samples showed the protocol efficiency in detecting total antioxidant capacity. In the test with microcystins a dose-dependent decrease (p<0.05) of antioxidant competence in gills and brain and an inverse result with liver samples were observed. The use of antioxidant defenses was efficient in avoiding oxidative damage, as the content of oxidized proteins was not altered. Data obtained show the potential of this new methodology to be used in ecotoxicological studies.