Acute exposure and biomarkers assessment in rainbow trout exposed to selected pharmaceuticals.

Pharmaceuticals released from municipal effluents discharges pose a risk to aquatic organisms. The toxicity of 5 pharmaceuticals with distinct therapeutic actions were assessed in rainbow trout: olanzapine (antipsychotic), erythromycin (antibiotic), mycophenoate (immunosuppression), pinaverium (anti-inflammatory) and trazodone (sedative). Juveniles were exposed to these drugs for 96 h at concentrations between 64 µg/L up to 40 mg/L to reach lethality. Survival was determined and a suite of biomarkers was analyzed for drug biotransformation, oxidative stress/damage and metabolic activity at sublethal concentrations. The data revealed the following toxicity: olanzapine >trazodone>mycophenolate>pinaverium∼erythromycin based on mortality. The data also revealed that toxicity was associated to mass, pKa and hydrophobicity and the following sublethal effects: GST, LPO and DNA strand breaks. Pharmaceuticals with lower molecular weight, physiological pKa, moderate hydrophobicity, low biotransformation and DNA strand breaks were generally more toxic to fish. However, this should be considered as a general guide in identifying toxic pharmaceuticals in non-target organisms.

Bioconcentration Assessment of Three Cationic Surfactants in Permanent Fish Cell Lines.

Cationic surfactants are used in many industrial processes and in consumer products with concurrent release into the aquatic environment, where they may accumulate in aquatic organisms to regulatoryly relevant thresholds. Here, we aimed to better understand the bioconcentration behavior of three selected cationic surfactants, namely N,N-dimethyldecylamine (T10), N-methyldodecylamine (S12), and N,N,N-trimethyltetradecylammonium cation (Q14), in the cells of fish liver (RTL-W1) and gill (RTgill-W1) cell lines. We conducted full mass balances for bioconcentration tests with the cell cultures, in which the medium, the cell surface, the cells themselves, and the plastic compartment were sampled and quantified for each surfactant by HPLC MS/MS. Accumulation in/to cells correlated with the surfactants' alkyl chain lengths and their membrane lipid-water partitioning coefficient, DMLW. Cell-derived bioconcentration factors (BCF) of T10 and S12 were within a factor of 3.5 to in vivo BCF obtained from the literature, while the cell-derived BCF values for Q14 were >100 times higher than the in vivo BCF. From our experiments, rainbow trout cell lines appear as a suitable conservative in vitro screening method for bioconcentration assessment of cationic surfactants and are promising for further testing.

Expanding non-invasive approaches for fish-health monitoring: A survey of the epidermal mucous metabolomes of phylogenetically diverse freshwater fish species.

There is a pressing need for more-holistic approaches to fisheries assessments along with growing demand to reduce the health impacts of sample collections. Metabolomic tools enable the use of sample matrices that can be collected with minimal impact on the organism (e.g., blood, urine, and mucus) and provide high-throughput, untargeted biochemical information without the requirement of a sequenced genome. These qualities make metabolomics ideal for monitoring a wide range of fish species, particularly those under protected status. In the current study, we surveyed the relative abundances of 120 endogenous metabolites in epidermal mucus across eight freshwater fish species belonging to seven phylogenetic orders. Principal component analysis was used to provide an overview of the data set, revealing strong interspecies relationships in the epidermal mucous metabolome. Normalized relative abundances of individual endogenous metabolites were then used to identify commonalities across multiple species, as well as those metabolites that showed notable species specificity. For example, taurine was measured in high relative abundance in the epidermal mucus of common carp (Cyprinus carpio), northern pike (Esox lucius), golden shiner (Notemigonus crysoleucas), rainbow trout (Oncorhynchus mykiss), and rainbow smelt (Osmerus mordax), whereas γ-amino butyric acid (GABA) exhibited a uniquely high relative abundance in flathead catfish (Pylodictis olivaris). Finally, hierarchical cluster analysis was used to evaluate species relatedness as characterized by both the epidermal mucous metabolome (phenotype) and genetic phylogeny (genotype). This comparison revealed species for which relatedness in the epidermal mucous metabolome composition closely aligns with phylogenetic relatedness (e.g., N. crysoleucas and C. carpio), as well as species for which these two measures are not well aligned (e.g., P. olivaris and Polyodon spathula). These, and other findings reported here, highlight novel areas for future research with fish, including development of epidermal mucous-based markers for non-invasive health monitoring, sex determination, and hypoxia tolerance.

In vitro toxicity assessment of polyethylene terephthalate and polyvinyl chloride microplastics using three cell lines from rainbow trout (Oncorhynchus mykiss).

The RTgill-W1 (gill), RTG-2 (gonad), and RTL-W1 (liver) cell lines derived from a freshwater fish rainbow trout (Oncorhynchus mykiss), were used to assess the toxicity of polyethylene terephthalate (PET) and two forms of polyvinyl chloride (PVC). Two size fractions (25-µm and 90-µm particles) were tested for all materials. The highest tested concentration was 1 mg/ml, corresponding to from 70 000 ± 9000 to 620 000 ± 57 000 particles/ml for 25-µm particles and from 2300 ± 100 to 11 000 ± 1000 particles/ml for 90-µm particles (depending on the material). Toxicity differences between commercial PVC dry blend powder and secondary microplastics created from a processed PVC were newly described. After a 24-h exposure, the cells were analyzed for changes in viability, 7-ethoxyresorufin-O-deethylase (EROD) activity, and reactive oxygen species (ROS) generation. In addition to the microplastic suspensions, leachates and particles remaining after leaching resuspended in fresh exposure medium were tested. The particles were subjected to leaching for 1, 8, and 15 days. The PVC dry blend (25 µm and 90 µm) and processed PVC (25 µm) increased ROS generation, to which leached chemicals appeared to be the major contributor. PVC dry blend caused substantially higher ROS induction than processed PVC, showing that the former is not suitable for toxicity testing, as it can produce different results from those of secondary PVC. The 90-µm PVC dry blend increased ROS generation only after prolonged leaching. PET did not induce any changes in ROS generation, and none of the tested polymers had any effect on viability or EROD activity. The importance of choosing realistic extraction procedures for microplastic toxicity experiments was emphasized. Conducting long-term experiments is crucial to detect possible environmentally relevant effects. In conclusion, the tested materials showed no acute toxicity to the cell lines.

Fish feeds supplemented with calcium-based buffering minerals decrease stomach acidity, increase the blood alkaline tide and cost more to digest.

Predatory fish in the wild consume whole prey including hard skeletal parts like shell and bone. Shell and bone are made up of the buffering minerals calcium carbonate (CaCO3) and calcium phosphate (Ca3(PO4)2). These minerals resist changes in pH, meaning they could have physiological consequences for gastric acidity, digestion and metabolism in fish. Using isocaloric diets supplemented with either CaCO3, Ca3(PO4)2 or CaCl2 as non-buffering control, we investigated the impacts of dietary buffering on the energetic cost of digestion (i.e. specific dynamic action or SDA), gastric pH, the postprandial blood alkalosis (the "alkaline tide") and growth in juvenile rainbow trout (Oncorhynchus mykiss). Increases in dietary buffering were significantly associated with increased stomach chyme pH, postprandial blood HCO3-, net base excretion, the total SDA and peak SDA but did not influence growth efficiency in a 21 day trial. This result shows that aspects of a meal that have no nutritional value can influence the physiological and energetic costs associated with digestion in fish, but that a reduction in the SDA will not always lead to improvements in growth efficiency. We discuss the broader implications of these findings for the gastrointestinal physiology of fishes, trade-offs in prey choice in the wild, anthropogenic warming and feed formulation in aquaculture.

Factors Affecting the Binding of Diltiazem to Rainbow Trout Plasma: Implications for the Risk Assessment of Pharmaceuticals in Aquatic Systems.

The accumulation of organic toxicants in fish plasma, and how they partition between the bound and unbound fraction once absorbed, are important metrics in models that seek to predict the risk of such contaminants in aquatic settings. Rapid equilibrium dialysis of diltiazem, an ionizable weak base and important human pharmaceutical contaminant of freshwaters, was conducted with rainbow trout (Oncorhynchus mykiss) plasma. The effect of fed state, fish sex, fish strain/size, and dialysis buffer pH on the binding of radiolabeled diltiazem (9 ng ml-1 ) was assessed. In fed fish, 24.6%-29.5% of diltiazem was free, unbound to plasma proteins. Although starvation of fish resulted in a decrease in plasma protein, the bound fraction of diltiazem remained relatively constant. Consequently, the protein-bound concentration of diltiazem increased with length of starvation. In general, rainbow trout strain was a significant factor affecting plasma binding, although the two strains tested also differed markedly in size. Dialysis buffer pH significantly influenced plasma binding, with a higher unbound diltiazem fraction at pH 6.8 than pH 8.0. These data indicate that empirical measures of plasma binding in fish are important for accurate risk assessment and that the physiological status of a fish is likely to impact its sensitivity to toxicants such as diltiazem. Environ Toxicol Chem 2022;41:3125-3133. © 2022 SETAC.

Investigation of long-term bisphenol A exposure on rainbow trout (Oncorhynchus mykiss): Hematological parameters, biochemical indicator, antioxidant activity, and histopathological examination.

As an industrial synthetic chemical, bisphenol A (BPA) has the potential to have physiologically and histologically adverse effects on aquatic organisms. BPA causes the reproductive disrupting of all vertebrates due to its degradation on endocrine system. Therefore, the effect of BPA on fish with high economic value is an important issue. This study focused weekly on long-term BPA exposure on rainbow trout (Oncorhynchus mykiss). Hematological, biochemical, antioxidant activity and histopathological examinations were performed on O. mykiss exposed to 1000 µg/L BPA concentration. Mortality was observed in the BPA group during the first three weeks. As a result of hematological studies, leukocyte count and hemoglobin in the BPA group were significantly higher in the first three weeks compared to the control group. Plasma cortisol level as a biochemical indicator showed a similar trend to leukocyte and hemoglobin. There was no significant difference between BPA and control groups in terms of superoxide dismutase and catalase. However, glutathione peroxidase activity in the BPA group was significantly lower than in the control group for four weeks. At the end of the study, many alterations were observed histologically in the gill tissues. While more intense hyperplasia and epithelial lifting were observed in the tissues in the BPA group compared to the control group, vacuolization, necrosis and hypertrophy were observed infrequently. In conclusion, this study argues that BPA causes negative effects on O. mykiss in terms of fish welfare and future study should be focused on its environmental adaptation with color preference patterns.

Integrative multi-biomarker approach on caged rainbow trout: A biomonitoring tool for wastewater treatment plant effluents toxicity assessment.

The complex mixtures of contaminants released in wastewater treatment plant (WWTP) effluents are a major source of pollution for aquatic ecosystems. The present work aimed to assess the environmental risk posed by WWTP effluents by applying a multi-biomarker approach on caged rainbow trout (Oncorhynchus mykiss) juveniles. Fish were caged upstream and downstream of a WWTP for 21 days. To evaluate fish health, biomarkers representing immune, reproductive, nervous, detoxification, and antioxidant functions were assayed. Biomarker responses were then synthesized using an Integrated Biomarker Response (IBR) index. The IBR highlighted similar response patterns for the upstream and downstream sites. Caged juvenile females showed increased activities of innate immune parameters (lysozyme and complement), histological lesions and reduced glycogen content in the hepatic tissue, and higher muscle cholinergic metabolism. However, the intensity of the observed effects was more severe downstream of the WWTP. The present results suggest that the constitutive pollution level of the Meuse River measured upstream from the studied WWTP can have deleterious effects on fish health condition, which are exacerbated by the exposure to WWTP effluents. Our results infer that the application of IBR index is a promising tool to apply with active biomonitoring approaches as it provides comprehensive information about the biological effects caused by point source pollution such as WWTP, but also by the constitutive pollutions levels encountered in the receiving environment.

P-glycoprotein induction and its energetic costs in rainbow trout (Oncorhynchus mykiss).

Biological organisms are constantly challenged by xenobiotics and have evolved mechanisms to reduce, neutralize, or repair toxic outcomes. The various chemical defenses all utilize energy, but their specific costs and impacts on energy budgets are currently unknown. In this study, the energetic costs associated with the induction and substrate transport of the efflux transporter P-glycoprotein (P-gp [ABCB1, MDR1]) were examined in rainbow trout. An intraperitoneal injection of the P-gp inducer clotrimazole (0, 0.1, 1.0, and 10 mg/kg) increased P-gp activity (as measured by a competitive rhodamine 123 transport assay in hepatocytes) in a dose-dependent manner reaching a maximum induction of 2.8-fold. Maximum P-gp induction occurred at 50 h post-administration with the highest dose; significant induction of P-gp activity remained elevated over constitutive values until the last sampling time point (168 h). In vitro measurements of hepatocyte respiration indicated that basal P-gp activity transporting R123 as a substrate did not significantly increase respiration rates (range 18.0 to 23.2 ng O2/min/106 cells); however, following the induction of P-gp by clotrimazole and exposure to the P-gp substrate R123, respiration rates increased significantly (3.52-fold) over baseline values. Using whole animal respirometry, it was shown that respiration rates in fish exposed to R123 only or induced with clotrimazole were not different from controls (range 1.2 to 2.1 mg O2/kg/min); however, respiration rates were significantly increased in fish with induced P-gp levels and also exposed to R123. This work indicates that basal and induced levels of P-gp activity do not incur significant energetic costs to fish; however, upon induction of P-gp and concomitant substrate exposures, energetic costs can increase and could pose challenges to organisms facing limited energy resources.

Assessment of oxidative DNA damage, oxidative stress responses and histopathological alterations in gill and liver tissues of Oncorhynchus mykiss treated with linuron.

We investigated changes in 8-hydroxy-2-deoxyguanosine (8-OHdG) activity which is a product of oxidative DNA damage, histopathological changes and antioxidant responses in liver and gill tissues of rainbow trout, following a 21-day exposure to three different concentrations of linuron (30 µg/L, 120 µg/L and 240 µg/L). Our results indicated that linuron concentrations caused an increase in LPO levels of liver and gill tissues (p < 0.05). While linuron induced both increases and decreases in GSH levels and SOD activity, CAT activity was decreased by all concentrations of linuron (p < 0.05). The immunopositivity of 8-OHdG was detected in the hepatocytes of liver and in the epithelial and chloride cells of the secondary lamellae of the gill tissues. Our results suggested that linuron could cause oxidative DNA damage by causing an increase in 8-OHdG activity in tissues, and it induces histopathological damage and alterations in the antioxidant parameters of the tissues.

Comparison of Alternative Methods for Bioaccumulation Assessment: Scope and Limitations of In Vitro Depletion Assays with Rainbow Trout and Bioconcentration Tests in the Freshwater Amphipod Hyalella azteca.

Bioaccumulation assessment predominantly relies on the bioconcentration factor (BCF) as the sole decisive metric. The test guideline 305 by the Organisation for Economic Co-operation and Development (OECD) provides the standard procedure for deriving this in vivo fish BCF, which is not only expensive and labor-intensive, but also requires many animals. Accordingly, there is a great need for and interest in alternative methods that can help to reduce, replace, and refine vertebrate tests, as described in the 3R principles. Two alternative approaches have been developed: the bioconcentration test with the freshwater amphipod Hyalella azteca and the OECD test guideline 319 which provides a method to determine experimentally derived in vitro metabolism rates that can then be incorporated into in silico prediction models for rainbow trout BCF calculation. In the present study both alternative methods were applied to 5 substances of different physicochemical characteristics. The results were compared with literature values of fish in vivo BCFs and additional BCFs obtained with the alternative methods, if available. Potential differences between the results of the test methods are discussed utilizing information such as in vivo metabolism rates. The currently available data set suggests that these 2 alternative methods pose promising alternatives to predict bioaccumulation in fish, although defined applicability domains have yet to be determined. Environ Toxicol Chem 2020;39:1813-1825. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

The concentration of potentially toxic elements (PTEs) in muscle tissue of farmed Iranian rainbow trout (Oncorhynchus mykiss), feed, and water samples collected from the west of Iran: a risk assessment study.

The pollution of the environment by potentially toxic elements (PTEs) is one of the most important raised concerns. Therefore, the current investigation was devoted to measuring the concentration of lead (Pb), cadmium (Cd), elemental mercury (Hg), nickel (Ni), iron (Fe), zinc (Zn), and copper (Cu) in muscle tissue of farmed rainbow trout (n = 30) as well as their feed (n = 15) and water (n = 15) samples collected from farms (Hamadan Province, Iran) by the aid of an inductively coupled plasma atomic emission spectrometer (ICP-OES). Also, the associated risk for human and biomagnification factor (BMF) and bioconcentration factor (BCF) for PTEs in the fish muscle through feed and water were calculated. The mean concentration of Pb, Cd, Hg, Ni, Fe, Zn, and Cu in rainbow trout muscle was reported as 0.056 ± 0.040 µg g-1 wet weight,

Toxicity assessment of pollutants sorbed on environmental sample microplastics collected on beaches: Part I-adverse effects on fish cell line.

Microplastics (MPs), are tiny plastic fragments from 1 µm to 5 mm generally found in the aquatic environment which can be easily ingested by organisms and may cause chronic physical but also toxicological effects. Toxicological assays on fish cell lines are commonly used as an alternative tool to provide fast and reliable assessment of the toxic and ecotoxic properties of chemicals or mixtures. Rainbow trout liver cell line (RTLW-1) was used to evaluate the toxicity of pollutants sorbed to MPs sampled in sandy beaches from different islands around the world during the first Race for Water Odyssey in 2015. The collected MPs were analyzed for polymer composition and associated persistent organic pollutants: polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT). In addition, DMSO-extracts from virgin MPs, MPs artificially coated with B[a]P and environmental MPs were analyzed with different bioassays: MTT reduction assay (MTT), ethoxyresorufin-O-deethylase (EROD) assay and comet assay. Microplastics from sand beaches were dominated by polyethylene, followed by polypropylene fragments with variable proportions. Organic pollutants found on plastic from beach sampling was PAHs (2-71 ng g-1). Samples from Bermuda (Somerset Long Bay) and Hawaii (Makapu'u) showed the highest concentration of PAHs and DDT respectively. No toxicity was observed for virgin microplastics. No cytotoxicity was observed on cells exposed to MP extract. However, EROD activity was induced and differently modulated depending on the MPs locations suggesting presence of different pollutants or additives in extract. DNA damage was observed after exposure to four microplastics samples on the six tested. Modification of EROD activity level and DNA damage rate highlight MPs extract toxicity on fish cell line.

Proactive coping style in early emerging rainbow trout carries a metabolic cost with no apparent return.

The timing with which salmonid larvae emerge from their gravel nests is thought to be correlated with a particular suite of behavioural and physiological traits that correspond to the stress coping style of the individual. Among these traits, aggressiveness, dominance and resilience to stress, are potentially interesting to exploit in aquaculture production. In the present study a series of experiments were performed, with the purpose of characterising behavioural, metabolic and production related traits in rainbow trout juveniles from different emergence fractions. Newly hatched rainbow trout were sorted according to their emergence time from an artificial redd. The early, middle, and late fractions were retained and assessed for their physiological response to stress, growth performance, metabolism, fasting tolerance, and potential for compensatory growth. The early emerging fraction showed proactive behavioural traits; they were faster to reappear following startling, showed a reduced cortisol response following stress, and a reduced metabolic cost of recovery. Emergence time was not correlated with any differences in standard or maximum metabolic rates, but was however, correlated with higher routine metabolic rates, as demonstrated by significantly bigger weight losses during fasting in the early emerging group. Growth rates and feed conversion efficiencies were not significantly different when fish were co-habitated under a restrictive feeding regime, suggesting that early emerging fish are not able to monopolise food resources. The intermediate emerging group, which makes up the bulk of a population and is often ignored, appears to possess the best growth performance traits, possibly because they do not expend excessive energy on dominance behaviour such as the early emerging group, while they are also not overly timid or stress prone such as the late emerging group.

Concentration dependence of in vitro biotransformation rates of hydrophobic organic sunscreen agents in rainbow trout S9 fractions: Implications for bioaccumulation assessment.

In vitro biotransformation studies were performed to support the bioaccumulation assessment of 3 hydrophobic organic ultraviolet filters (UVFs), 4-methylbenzylidene camphor (4-MBC), 2-ethylhexyl-4-methoxycinnamate (EHMC), and octocrylene. In vitro depletion rate constants (kdep ) were determined for each UVF using rainbow trout liver S9 fractions. Incubations performed with and without added cofactors showed complete (4-MBC) or partial (EHMC and octocrylene) dependence of kdep on addition of the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH), suggesting that hydrolysis of EHMC and octocrylene by NADPH-independent enzymes (e.g., carboxylesterases) is an important metabolic route. The concentration dependence of kdep was then evaluated to estimate Michaelis-Menten parameters (KM and Vmax ) for each UVF. Measured kdep values were then extrapolated to apparent whole-body biotransformation rate constants using an in vitro-in vivo extrapolation (IVIVE) model. Bioconcentration factors (BCFs) calculated from kdep values measured at concentrations well below KM were closer to empirical BCFs than those calculated from kdep measured at higher test concentrations. Modeled BCFs were sensitive to in vitro binding assumptions employed in the IVIVE model, highlighting the need for further characterization of chemical binding effects on hepatic clearance. The results suggest that the tested UVFs are unlikely to accumulate to levels exceeding the European Union Registration, Evaluation, Authorisation, and Restriction regulation criterion for bioaccumulative substances (BCF > 2000 L kg-1 ). However, consideration of appropriate in vitro test concentrations and binding correction factors are important when IVIVE methods are used to refine modeled BCFs. Environ Toxicol Chem 2019;38:548-560. © 2018 SETAC.

Assessment of acute and chronic toxicity of unweathered and weathered diluted bitumen to freshwater fish and invertebrates.

This paper presents the results of two different studies investigating the acute and chronic toxicity of two blends of diluted bitumens ("dilbit") and weathered dilbit on freshwater fish and invertebrates after exposure to different concentrations of physically-dispersed (water accommodated fraction; WAF) and chemically-dispersed (chemically-enhanced WAF; CEWAF). The first study evaluated the acute and chronic toxicity of weathered, unweathered and dispersed Access Western Blend (AWB) dilbit on Fathead minnow (Pimephales promelas). In the second study, acute and chronic toxicity of weathered and unweathered Cold Lake Blend (CLB) dilbit was assessed on Rainbow trout (Oncorhynchus mykiss), and two invertebrate species, Daphnia (Daphnia magna) and Ceriodaphnia (Ceriodaphnia dubia). For Fathead minnow, unweathered AWB WAF demonstrated a significantly higher acute toxicity (LC50-96 h = 0.628 g/L) compared to the weathered AWB WAF (LC50-96 h = 2.06 g/L). Chronic toxicity tests showed that Fathead minnow lethality was also higher for unweathered AWB (LC50-7 d = 0.593 g/L) compared to the weathered AWB (LC50-7 d = 1.31 g/L) whereas larval growth toxicity was lower for unweathered AWB (IC25-7 d=0.312 g/L) compared to the weathered dilbit (IC25-7 d = 0.096 g/L). Rainbow trout exposed to unweathered CLB demonstrated a significantly higher toxicity (LC50-96 h = 5.66 g/L) compared to the weathered CLB (LC50 > 18 g/L). Lethality (LC50 = 6.43 g/L) was observed in Ceriodaphnia exposed to the CLB WAF while no mortality was observed with the weathered CLB. The reproductive effects on Ceriodaphnia were greater with the CLB (IC25 <1.0) than with the weathered CLB (IC25 = 3.99 g/L). Volatile organic compounds (VOC), polycyclic aromatic hydrocarbons (PAH) and total petroleum hydrocarbons (TPH) increased as the dilbit CLB and AWB WAF concentrations increased. The total VOC and the total PAHs (including the alkylated PAHs) presented similar concentrations in the weathered and unweathered AWB WAF. These compounds seem therefore not be affected by the AWB WAF weathering process. However, VOC and PAH concentrations decreased significantly in the Rainbow trout and Ceriodaphnia toxicity tests using the weathered CLB WAF. VOC and PAH concentrations were also lower in the weathered AWB CEWAF used for Fathead minnow chronic study.

Formation of PFAAs in fish through biotransformation: A PBPK approach.

A physiologically-based pharmacokinetic (PBPK) model for perfluorinated alkyl acids (PFAAs) in rainbow trout has been updated to include formation of perfluorooctanoic acid (PFOA) from the biotransformation of 8:2 fluorotelomer carboxylic acid (8:2 FTCA). The updated model is dynamic and simulates both uptake and depuration phases. Two empirical studies are used to parameterize and test the model. In the first case, parameters related to fecal elimination and protein binding were optimized. In the second case, parameters were sourced either from literature or from optimized values based on the first study to test model performance. Optimization of parameters resulted in a decrease in the difference between experimental data and simulation results by 57 and 23 percent for the first and the second case, respectively, compared to the original case. Sensitivity analysis was performed to identify important parameters, and uncertainty in model prediction propagated by these parameters was assessed using Monte Carlo analysis. For each case, 80 and 89 percent, respectively, of median predicted values were within the limits of experimental error when comparing simulated and experimental data. This is the first toxicokinetic model that incorporates biotransformation of PFAA precursors and simultaneously predicts the distribution of the precursor and metabolite in different tissues. The model is mechanistic, and could be applied to simulate a variety of scenarios by using the organism-specific physiological properties compiled here with other chemical-specific parameters (e.g. protein interactions).

Assessment of 8-hydroxy-2-deoxyguanosine activity, gene expression and antioxidant enzyme activity on rainbow trout (Oncorhynchus mykiss) tissues exposed to biopesticide.

The goal of this study was to determinate toxicity mechanism of biopesticide with antioxidant enzymes parameters such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) and malondialdehyde (MDA) levels, oxidative DNA damage (8-hydroxy-2-deoxyguanosine (8-OHdG)), transcriptional changes of heat shock protein 70 (HSP70), and cytochromes P4501A (CYP1A), sod, cat, and gpx in liver and gill tissues of Oncorhynchus mykiss. For this aim, plant-based (natural pesticides, azadirachtin (AZA)) and synthetic pesticides (deltamethrin (DLM)) were exposed on the fish at different concentrations (0.0005 and 0.00025ppm of DLM; 0.24 and 0.12ppm of AZA) for 21 days. According to the results of the study, the activity of SOD, CAT and GPx decreased, but malondialdehyde (MDA) level and activity of 8-OHdG increased in the gill and liver of rainbow trout (p<0.05). Additionally sod, cat and gpx were down regulated; HSP70 and CYP1A were up regulated for transcriptional observation. The downwards regulation of antioxidant (sod, cat and gpx) and the upregulation of HSP70 and CYP1A was obvious with doses of AZA or DLM (p<0.05). The findings of this study suggest that biopesticide can cause biochemical and physiological effects in the fish gill and liver by causing enzyme inhibition, an increase in 8-OHdG levels and changes in both transcriptional parameters (sod, cat, gpx, HSP70 and CYP1A). We found that excessive doses of plant-based pesticide are nearly as toxic as chemical ones for aquatic organisms. Moreover, 8-OHdG, HSP70 and CYP1A used as a biomarker to determinate toxicity mechanism of biopesticide in aquatic environment.

Comparison of heavy metal levels of farmed and escaped farmed rainbow trout and health risk assessment associated with their consumption.

In this study, levels of ten metals (arsenic, cadmium, cobalt, chromium, copper, iron, manganese, nickel, lead, and zinc) in muscles of farmed and escaped farmed rainbow trout (Oncorhynchus mykiss) in the Keban Dam Reservoir (Turkey) were determined. Also, human health risks associated with their consumption were assessed. Of ten metals, only Co and Fe levels in escaped rainbow trout were significantly higher than those in farmed rainbow trout. The metal levels in farmed and escaped rainbow trout were below the maximum permissible limits. The estimated daily intake (EDI) of each metal in both farmed and escaped farmed rainbow trout was much lower than the respective tolerable daily intake (TDI). The target hazard quotient (THQ) values for individual metal and the total THQ values for combined metals were lower than 1 in both farmed and escaped rainbow trout, indicating no health risk for humans. The cancer risk (CR) values estimated for inorganic As in both farmed and escaped rainbow trout indicated low carcinogenic risk to the consumers. According to the maximum allowable monthly consumption limits (CRmm), adults may safely consume 24 meals of farmed rainbow trout per month or 39 meals of escaped rainbow trout per month, with minimal adverse carcinogenic and non-carcinogenic health effects. This study revealed that the risk from consuming farmed and escaped farmed rainbow trout in the Keban Dam Reservoir due to these trace elements is minimal.

Increased mitochondrial coupling and anaerobic capacity minimizes aerobic costs of trout in the sea.

Anadromy is a distinctive life-history strategy in fishes that has evolved independently many times. In an evolutionary context, the benefits of anadromy for a species or population must outweigh the costs and risks associated with the habitat switch. The migration of fish across the freshwater-ocean boundary coincides with potentially energetically costly osmoregulatory modifications occurring at numerous levels of biological organization. By integrating whole animal and sub-cellular metabolic measurements, this study presents significant findings demonstrating how an anadromous salmonid (i.e. rainbow trout, Oncorhynchus mykiss) is able to transform from a hyper- to hypo-osmoregulatory state without incurring significant increases in whole animal oxygen consumption rate. Instead, underlying metabolic mechanisms that fuel the osmoregulatory machinery at the organ level (i.e. intestine) are modulated, as mitochondrial coupling and anaerobic metabolism are increased to satisfy the elevated energetic demands. This may have positive implications for the relative fitness of the migrating individual, as aerobic capacity may be maintained for locomotion (i.e. foraging and predator avoidance) and growth. Furthermore, the ability to modulate mitochondrial metabolism in order to maintain osmotic balance suggests that mitochondria of anadromous fish may have been a key target for natural selection, driving species adaptations to different aquatic environments.