Exposure to pollutants present in Iguaçu River Southern Brazil affect the health of Oreochromis niloticus (Linnaeus, 1758): Assessment histological, genotoxic and biochemical.

Urban sewage is a source of major contamination in aquatic systems and contributes to environmental and human health disturbances. This study investigates the effects of sewage-polluted waters from Iguaçu River on the health of juvenile Oreochromis niloticus. Two hundred four specimens were exposed to riverine water in four groups: no diluted, 25 and 50 % diluted water and a control group without tested water for 72 days. Biological samples were obtained for histopathological, neurotoxicity, antioxidant defenses, genotoxicity, metallothionines expression and polycyclic aromatic hydrocarbons (PAHs) metabolites. The results showed histopathological alterations in liver and gills, genotoxic alteration in erythrocytes, reduction of acetylcholinesterase activity in brain and muscle, activation of antioxidant defenses in the liver, recruitment of metals by metallothionein and the detection of PAHs metabolites in bile. These results demonstrate that juveniles of O. niloticus are susceptible to Iguaçu River exposure water and they can be used as indicator of water quality.

Comparative Assessment of Genotoxic Impacts Induced by Zinc Bulk- and Nano-Particles in Nile tilapia, Oreochromis niloticus.

Fish were separately exposed to 1/2 LC50/96 h values of bulk-Zn and nano-Zn for 7, 14, and 28 days. The induction of micronuclei (MN) and other eight nuclear abnormalities in erythrocytes showed marked time and size dependence. The frequencies of all nuclear anomalies were progressively elevated (p < 0.05) with increasing the time of exposure to both bulk-Zn and nano-Zn. Throughout the study periods, fish exposed to nano-Zn showed the maximum elevation in all studied nuclear anomalies. Based on the fragmented DNA values, both Zn forms induced tissue-specific DNA damage as following gills > liver > muscles. Moreover, nano-Zn exposed groups revealed a maximum percentage of DNA damage among all studied groups, especially after 14 days. The percentage of DNA damage was decreased in all tissues on the 28th day, which reflected the presence of an effective repair mechanism. Finally, nano-Zn exhibited more genotoxic effects than that of its bulk counterparts.

Health risk assessment of heavy metal accumulation in the Buriganga and Turag River systems for Puntius ticto, Heteropneustes fossilis, and Channa punctatus.

This study aimed to assess the effects of major ecotoxic heavy metals accumulated in the Buriganga and Turag River systems on the liver, kidney, intestine, and muscle of common edible fish species Puntius ticto, Heteropneustes fossilis, and Channa punctatus and determine the associated health risks. K was the predominant and reported as a major element. A large concentration of Zn was detected in diverse organs of the three edible fishes compared with other metals. Overall, trace metal analysis indicated that all organs (especially the liver and kidney) were under extreme threat because the maximum permissible limit set by different international health organizations was exceeded. The target hazard quotient and target cancer risk due to the trace metal content were the largest for P. ticto. Thus, excessive intake of P. ticto from the rivers Buriganga and Turag could result in chronic risks associated with long-term exposure to contaminants. Histopathological investigations revealed the first detectable indicators of infection and findings of long-term injury in cells, tissues, and organs. Histopathological changes in various tissue structures of fish functioned as key pointers of connection to pollutants, and definite infections and lesion types were established based on biotic pointers of toxic/carcinogenic effects. The analysis of histopathological alterations is a controlling integrative device used to assess pollutants in the environment.

Assessment of the heavy metal accumulation in the Blue Swimmer Crab (Portunus pelagicus), northern Bay of Bengal: Role of salinity.

This is the first report to assay the heavy metal accumulation in Blue Swimmer Crab in the northern Bay of Bengal. The present study finds that copper (Cu) concentration though has been the highest compared to the other metals, particularly in the gill. Cadmium (Cd) and lead (Pb) have high affinity to be concentrated in Hepatopancreas all through the year. All the metal concentrations were higher during monsoon season (comparatively low salinity) than non-monsoon (high salinity) in the different body parts of Blue swimmer crab i.e. the inverse relationship between accumulation of heavy metals and sea water salinity has been shown. Present study revealed that salinity having the role to accumulate metal in different body parts of Blue Swimmer Crab. In future, microcosm experiments should be done to test the effect of frequent fluctuations of ambient salinity, such as is usual in different seasons in the present study area.

Severe and protracted cholestasis in 44 young men taking bodybuilding supplements: assessment of genetic, clinical and chemical risk factors.

BACKGROUND: Bodybuilding supplements can cause a profound cholestatic syndrome. AIM: To describe the drug-Induced liver injury network's experience with liver injury due to bodybuilding supplements. METHODS: Liver injury pattern, severity and outcomes, potential genetic associations, and exposure to anabolic steroids by product analysis were analysed in prospectively enrolled subjects with bodybuilding supplement-induced liver injury with causality scores of probable or higher. RESULTS: Forty-four males (mean age 33 years) developed liver injury with a median latency of 73 days. Forty-one per cent presented with hepatocellular pattern of liver injury as defined by the R > 5 ([Fold elevation of ALT] ÷ [Fold elevation of Alk Phos] (mean, range = 6.4, 0.5-31.4, n = 42) despite all presenting with clinical features of cholestatic liver injury (100% with jaundice and 84% with pruritus). Liver biopsy (59% of subjects) demonstrated a mild hepatitis and profound cholestasis in most without bile duct injury, loss or fibrosis. Seventy-one per cent were hospitalised, and none died or required liver transplantation. In some, chemical analysis revealed anabolic steroid controlled substances not listed on the label. No enrichment of genetic variants associated with cholestatic syndromes was found, although mutations in ABCB11 (present in up to 20%) were significantly different than in ethnically matched controls. CONCLUSIONS: Patients with bodybuilding supplements liver injury uniformly presented with cholestatic injury, which slowly resolved. The ingested products often contained anabolic steroids not identified on the label, and no enrichment in genetic variants was found, indicating a need for additional studies.

Assessment of the acute toxicity of chlorpyrifos and cypermethrin to Heteropneustes fossilis and their impact on acetylcholinesterase activity.

In the present study, the acute toxicity of chlorpyrifos (an organophosphate, OP) and cypermethrin (a pyrethroid) pesticides was estimated for 96 h in Heteropneustes fossilis. The LC50 for chlorpyrifos (CPF) and cypermethrin was found to be 1.90 mg/L and 0.085 mg/L, respectively. The acetylcholinesterase (AChE, EC 3.1.1.7) activity in Heteropneustes fossilis exposed to both the insecticides was assayed in brain, muscle and gills. In general, tissue specific as well as dose-dependent decrease in the AChE activity was exhibited by both pesticides. In response to the increasing concentrations of chlorpyrifos and cypermethrin as well, a significant decrease in the activity of AChE was found in brain while muscle and gills exhibited lesser inhibition. Thus, the brain was the main target organ for both insecticides, followed by muscle and gills. Between the two pesticides chlorpyrifos acted as more potent AChE inhibitor than cypermethrin since more intense changes in behavioral pattern was observed with the chlorpyrifos. These changes indicate that the effects of these pesticides are at neural as well as neuromuscular level.

Enteric nervous system analyses: New biomarkers for environmental quality assessment.

The gastrointestinal tract (GIT) of fish is a target of contaminants since it can absorb these substances. We evaluated the morphophysiological alterations in the GIT of Sphoeroides testudineus collected in two estuaries presenting differences in their environmental quality (NIA and IA). The intestine was analyzed for histological and neuronal changes; liver and gills for biochemical markers; muscle tissues for neurotoxicity and peripheral blood for genotoxic damage. The results showed alterations in the GIT of the animals collected in the IA, such as muscle tunica and goblet cell density reduction, increased intraepithelial lymphocytes density and changes in neuronal density. Furthermore, changes were observed in MTs and LPO in the gills. Thus, we suggest that TGI is functioning as a barrier that responds to ingested contaminants, in order to reduce their absorption and translocation. Thus, alterations in morphophysiological and enteric neurons in S. testudineus can be used as biomarkers of environmental contamination.

Assessment of arsenic trioxide toxicity on cock muscular tissue: alterations of oxidative damage parameters, inflammatory cytokines and heat shock proteins.

To evaluate the toxicity of arsenic trioxide (As2O3) in the muscular tissues (wing, thigh and pectoral) of birds, 72 one-day-old Hy-line cocks were selected and randomly divided into four groups. They were fed either a commercial diet or an arsenic-supplemented diet containing 7.5, 15 or 30 mg/kg As2O3. The experiment lasted for 90 days and the samples of muscular tissues were collected at 30, 60 and 90 days. The results showed that As2O3 exposure significantly lowered the activities of antioxidant enzymes (catalase (CAT), glutathione peroxidase (GSH-Px)) and inhibition ability of hydroxyl radicals (OH) and increased the malondialdehyde (MDA) contents. Furthermore, the mRNA levels of inflammatory cytokines (tumor necrosis factor-α (TNF-α), nuclear factor-kappa B (NF-κB), cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS), prostaglandin E synthase (PTGEs)) and heat shock proteins (HSPs) in muscular tissue were significantly upregulated in the As2O3 exposure groups. The results indicated that As2O3 exposure resulted in oxidative damage, induced the inflammatory response, and influenced the mRNA levels of HSPs in muscular tissue of cocks. Additionally, the results suggested that HSPs possibly resisted due to the As2O3 exposure-induced oxidative stress and inflammatory response, which provided a favorable environment and played protective roles in the muscular tissues of cocks. The information presented in this study is helpful to understand the mechanism of As2O3 toxicity in bird muscular tissues.

Assessment of immune status of yellowfin seabream (Acanthopagrus latus) during short term exposure to phenanthrene.

The aim of the present investigation was to assess the immune status in yellowfin seabream (Acanthopagrus latus) exposed to different concentrations of phenanthrene (Phe) for 14days. In addition, the Phe accumulation in the fish muscle was measured during the experiment. Fish were injected with different concentrations (0, 2, 20 and 40mg/kg) of Phe and samples were taken from tissue and blood of fish 1, 4, 7 and 14days after injection. Exposure of fish to Phe caused a significant decrease in white blood cells, C3 and C4 levels, lysosomal membrane stability, lysozyme activity after 4days and antibacterial activity after 7days of the experiment. In contrast, cortisol level significantly increased after 4days. The concentration of Phe in fish muscle increased rapidly after 4days. The main tissue changes observed in the head kidney including increase in melanomacrophage centers (MMCs), empty spaces between cells and hemorrhage. The degree of tissue changes ranged from normal to moderate in Phe-treated fish. The size and number of MMCs in treated fish were significantly higher than control. In conclusion, Phe toxicity in yellowfin seabream can induce increased cortisol level, tissue changes and immune suppression.

The Growth-Promoting Effect of Dietary Nucleotides in Fish Is Associated with an Intestinal Microbiota-Mediated Reduction in Energy Expenditure.

Background: Nucleotides have been used as functional nutrients to improve the growth and health of animals, including fish. The mechanism involved in the growth-promotion effect of nucleotides is still unclear.Objective: We investigated the bioenergetic mechanism underlying the growth-promotion effect of nucleotides in zebrafish and the associated roles played by the intestinal microbiota.Methods: Larval zebrafish were fed a control or a 0.1% mixed nucleotides-supplemented diet for 2 wk. Standard metabolic rate, the minimal rate of energy expenditure by animals at rest, was evaluated by oxygen consumption with the use of a respirometer. The expressions of fasting-induced adipose factor (Fiaf), inflammatory cytokines, and genes involved in fatty acid (FA) oxidation were tested by quantitative reverse transcriptase-polymerase chain reaction. The intestinal microbiota from the nucleotide-fed fish (NT fish) or control fish was transferred to 3-d postfertilization germ-free zebrafish in which oxygen consumption and expression of cytokines and fiaf were evaluated.Results: Compared with controls, nucleotide supplementation at 0.1% increased the weight and energy gains of zebrafish by 10% and 25%, respectively (P < 0.01). Standard metabolic rate was 28% lower in NT fish than in controls (P < 0.001). Nucleotide supplementation downregulated the inflammatory tone in the head kidney of the fish. Moreover, NT fish had a 51% lower intestinal expression of fiaf than did controls (P < 0.05), which was consistent with decreased expression of key genes involved in FA oxidation [carnitine:palmitoyl transferase 1a (cpt1a) and medium-chain acyl coenzyme A dehydrogenase (mcad)] in liver and muscle. Germ-free zebrafish colonized with microbiota from NT fish had a 25% lower standard metabolic rate than did those colonized by control microbiota (P < 0.01), whereas direct nucleotide feeding of germ-free zebrafish did not affect standard metabolic rate relative to germ-free controls that were not fed nucleotides. Furthermore, germ-free zebrafish colonized with nucleotide microbiota exhibited downregulated inflammatory tone and 33% lower fiaf expression compared with their control microbiota-colonized counterparts.Conclusions: The growth-promoting effect of dietary nucleotides in zebrafish involves 2 intestinal microbiota-mediated mechanisms that result in reduced standard metabolic rate: 1) lower inflammatory tone and 2) reduced FA oxidation associated with increased microbial suppression of intestinal fiaf.

Assessment of chlorpyrifos toxic effects in zebrafish (Danio rerio) metabolism.

In this work the effect of chlorpyrifos exposure on metabolic profiles of zebrafish muscle was evaluated by liquid chromatography coupled to high resolution mass spectrometry. Different chemometric tools based on the selection of Regions of Interest and on Multivariate Curve-Resolution-Alternating Least Squares are proposed for the analysis of the complex data sets generated in the different exposure experiments. Analysis of Variance Simultaneous Component Analysis of changes on metabolite peak profile areas showed significant chlorpyrifos concentration and exposure time-dependent changes, clearly differentiating between exposed and non-exposed samples and between short (2 h) and long exposure times (6 h or 24 h). The changes observed in the concentrations of 50 muscle metabolites are indicative of induction of oxidative stress, of a general disruption of neurotransmitter metabolism, and of muscle exhaustion. These three effects are intimately related to the toxicity of chlorpyrifos.

The levels of mercury, methylmercury and selenium and the selenium health benefit value in grey-eel catfish (Plotosus canius) and giant mudskipper (Periophthalmodon schlosseri) from the Strait of Malacca.

The present study examined the concentrations of mercury (Hg), methylmercury (MeHg), and selenium (Se) in the multiple tissues of the Plotosus canius and Periophthalmodon schlosseri collected from the Strait of Malacca. The mean value in mg kg(-1) of Hg (P. canius: 0.34 ± 0.19; P. schlosseri: 0.32 ± 0.18) and MeHg in muscle (P. canius: 0.14 ± 0.11; P. schlosseri: 0.17 ± 0.11) were below the Codex general standard for contaminants and toxins in food and feed (CODEX STAN 193-1995), the Malaysian Food Regulation 1985 and the Japan Food Sanitation Law. For P. canius, the liver contained the highest concentrations of Hg (0.48 ± 0.07 mg kg(-1)) and MeHg (0.21 ± 0.00 mg kg(-1)), whereas for P. schlosseri, the gill contained the highest concentrations of Hg (0.36 ± 0.06 mg kg(-1)) and MeHg (0.21 ± 0.05 mg kg(-1)). The highest concentration of (80)Se (mg kg(-1)) was observed in the liver of P. canius (20.34 ± 5.68) and in the gastrointestinal tract (3.18 ± 0.42) of P. schlosseri. The selenium:mercury (Se:Hg) molar ratios were above 1 and the positive selenium health benefit value (HBVSe) suggesting the possible protective effects of Se against Hg toxicity. The estimate weekly intakes (EWIs) in µg kg(-1) body weight (bw) week(-1) of Hg (P. canius: 0.27; P. schlosseri: 0.15) and MeHg (P. canius: 0.11; P. schlosseri: 0.08) were found to be lower than the provisional tolerable weekly intake established by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Based on the calculated EWIs, P. canius, and P. schlosseri were found to be unlikely to cause mercury toxicity in human consumption.

Analysis of mercury and methylmercury concentrations, and selenium:mercury molar ratios for a toxicological assessment of sperm whales (Physeter macrocephalus) in the most recent stranding event along the Adriatic coast (Southern Italy, Mediterranean Sea).

Mass stranding of sperm whales (Physeter macrocephalus) is a rare event in the Mediterranean Sea. In September 2014, a pod of seven sperm whales became stranded along the Adriatic coast of Southern Italy. This is the seventh occurrence of this type since 1555 in this sea basin. Total concentrations of mercury (T-Hg), methylmercury (MeHg) and selenium (Se) were measured from brain, muscle, liver and kidney of three female sperm whales, which died in this event. Analyses showed considerable, age-dependent variations in Hg and Se concentrations in the different organs. The contamination levels of T-Hg in the liver (up to 200 mg kg(-1)) and brain (up to 21 mg kg(-1)) samples were markedly higher than those in the kidney and muscle samples. The liver and brain also showed the highest Se levels. Se:Hg molar ratios ⩾1 were observed in all the organs of the three sperm whales, suggesting that Se could protect the animals from Hg toxicity. The risk of Hg-associated neurotoxicity was assessed by comparing our values to thresholds set for neurotoxicity in mammals, and the role of Se in the detoxification process of T-Hg/MeHg is discussed herein.

Ecotoxicological assessment of tannery effluent using guppy fish (Poecilia reticulata) as an experimental model: a biomarker study.

Tannery wastewater in the East Calcutta Wetlands (a Ramsar site of West Bengal; number 1208) exerts adverse effects on commercial fish production and subsequently affects humans. The present study was conducted to investigate acute and chronic toxicity of tannery effluent on a fish biosystem by examining oxidative stress enzyme expression in different organs including liver, gills, and muscle following exposure. Phosphatases, both alkaline phosphatase and acid phosphatase, and antioxidant superoxide dismutase and catalase enzyme activities were determined in guppy fish (Poecilia reticulata) exposed to sublethal concentrations of composite tannery effluent. Data demonstrated that tannery effluent was capable of interfering with metabolic processes of fish by altering stress enzyme activities in fish organs, resulting in cellular injury. Data suggest that elevated activities of stress enzymes in fish upon exposure to environmental pollutants may serve as important biomarkers for oxidative stress.

Toxicity of cadmium and lead in Gallus gallus domesticus assessment of body weight and metal content in tissues after metal dietary supplements.

The influence of dietary cadmium on the accumulation and effects of dietary lead, examined in chicken. This experiment was conducted to investigate the toxic effects of dietary Cd and Pb on chick's body weight and organ, content of the tissues of these two metals was also detected. One day age chicks of Gallus gallus domesticus fed diet supplemented with 25, 50, 100 ppm of Cd, second group exposure to 300, 500, 1000 ppm of Pb in feed daily during 4 weeks. The control groups were fed without supplementation of metals. The concentrations of Cd and Pb resulted in increased of Cd and Pb content in liver, gizzard and muscle. While Cd 100 ppm and Pb 1000 ppm were increased metals content in feather. Body weight of chicks was not influenced by Cd treatment. In contrary Pb treatment was significantly (p < 0.05) decreased body weight of chicks after dietary treatment. On the other hand, Liver weigh in chicks was significantly (p < 0.05) decreased after Cd and Pb treatments.

Assessment of the contact toxicity of methyl palmitate on Tetranychus viennensis (Acari: Tetranychidae).

Previous work demonstrated that methyl palmitate possesses acaricidal activity against Tetranychus viennensis Boisduval (Acari: Tetranychidae) via an unknown mechanism. Here, the symptoms of methyl palmitate toxicity to T. viennensi were studied to determine the acaricidal mechanism of action of this fatty acid methyl ester. Methyl palmitate caused concentration-dependent mortality of T. viennensis, with a moderate concentration (5 mg/ml) eliciting excitement and premature oviposition without spinning shortly after exposure. Tremors of the appendages were subsequently observed, followed by quiescence after approximately 5 h. Mites developed dorsal fluid exosmosis at 15-20 h posttreatment with reduced egg production, followed shortly thereafter by death. Some typical neurotoxic symptoms such as excitement and convulsions were observed in methyl palmitate-exposed mites, suggesting that methyl palmitate may be a neurotoxin. Compared with other neurotoxic acaricides, methyl palmitate poisoning is a slow process in mites. Transmission electron microscopy revealed serious ultrastructural damage in response to 5 mg/ml methyl palmitate exposure. Autolysis of membranous structures was also observed, especially in the mitochondria, suggesting a novel mode of action for methyl palmitate-induced toxicity.

Assessment of neuromuscular dysfunction during poisoning by organophosphorus compounds.

Dysfunction of respiratory muscles is a life-threatening complication in poisoning by organophosphorus compounds (OPs). It is both of central and peripheral origin due to impaired cholinergic signalling upon inhibition of acetylcholinesterase (AChE). The dysfunction at neuromuscular synapses is not amenable to anticholinergics and remains a therapeutic challenge. Thus, a clear understanding of the distinct mechanisms occurring at neuromuscular synapses is decisive for the development and improvement of therapeutic strategies, particularly with nerve agent poisoning, where clinical studies are prevented by ethical considerations. Using red blood cell AChE, the kinetics of OP induced inhibition, aging, and spontaneous and oxime-induced reactivation have been elucidated. In a dynamically working in vitro model with real-time determination of membrane-bound AChE, it was shown that the kinetic constants derived from erythrocyte AChE are comparable to muscle AChE in a given species. To assess, whether kinetic considerations of AChE activity are relevant for the neuromuscular function, organotypic spinal cord-skeletal muscle cocultures have been established. In this model neostigmine and VX affected neuromuscular transmission as anticipated from their known actions on AChE. Also oxime-induced restoration of the neuromuscular transmission was observed. These findings were confirmed by functional studies on diaphragm muscles of various species with determination of muscle force generation upon phrenic nerve or indirect electrical field stimulation techniques. Investigations with human intercostal muscles are in progress to assess the conditions in human tissue. The results obtained with paraoxon favourably correlate with data from clinical findings of parathion-poisoned patients where the correlation of neuromuscular transmission with the activity of erythrocyte AChE could be established. In conclusion, a variety of methods are available to follow the microscopic reactions occurring at the synaptic level. Due to the lack of clinical data with different OPs, e.g. nerve agents, well designed animal experiments, reflecting the human situation as close as possible, are indispensable for the development of new drugs against the deleterious OP effects.

Botulinum neurotoxin: the ugly duckling.

This review presents a brief account of the most significant biological effects and clinical applications of botulinum neurotoxins, in a way comprehensive even for casual readers who are not familiar with the subject. The most toxic known substances in botulinum neurotoxins are polypeptides naturally synthesized by bacteria of the genus Clostridium. These polypeptides inhibit acetylcholine release at neuromuscular junctions, thus causing muscle paralysis involving both somatic and autonomic innervation. There is substantial evidence that this muscle-paralyzing feature of botulinum neurotoxins is useful for their beneficial influence on more than 50 pathological conditions such as spastic paralysis, cerebral palsy, focal dystonia, essential tremor, headache, incontinence and a variety of cosmetic interventions. Injection of adequate quantities of botulinum toxins in spastic muscles is considered as a highly hopeful procedure for the treatment of people who suffer from dystonia, cerebral palsy or have experienced a stroke. So far, numerous and reliable studies have established the safety and efficacy of botulinum neurotoxins and advocate wider clinical therapeutic and cosmetic applications.

Assessment of oxidative stress and histopathology in juvenile northern pike (Esox lucius) inhabiting lakes downstream of a uranium mill.

Lakes receiving effluent from the Key Lake uranium mill in northern Saskatchewan contain elevated trace metals, some of which are associated with increased reactive oxygen species (ROS) in cells and tissues causing oxidative stress. The potential for oxidative stress was assessed in juvenile (age 1+) northern pike (Esox lucius) collected from two exposure (high and low) and one reference lake near the Key Lake operation. The concentrations of total, reduced and oxidized glutathione and the ratio of oxidized to reduced glutathione in liver and kidney did not differ significantly among pike collected from exposure and reference lakes, with the exception of low exposure pike kidney that had significantly greater oxidized glutathione and ratio of oxidized to reduced glutathione. The concentrations of by-products of lipid peroxidation (malondialdehyde and 4-hydroxyalkenal) were significantly greater in kidney of pike collected from the reference lake compared to both exposure lakes. The activity of the antioxidant enzyme glutathione peroxidase in liver was greater in pike collected from the high exposure lake compared to the reference lake. Histopathological evaluations revealed greater pathology in reference lake pike as indicated by a greater number of pyknotic and fragmented nuclei and dilated tubules as well as a thickening of Bowman's capsule in kidney, and as a thickening of the primary filament epithelial padding in gills. In liver, hepatocyte morphology, including transsectional area and degree of vacuolation, differed among lakes without any clear signs of pathology. Trace metal analyses of muscle showed that eight elements (arsenic, cobalt, copper, iron, molybdenum, selenium, thallium, and uranium) were significantly elevated in pike collected from both exposure lakes compared to reference. These results provide only limited evidence of oxidative stress in exposure pike tissues and no evidence of histopathology despite indications that trace metals, most notably arsenic and selenium, were bioaccumulating in tissue.