Freshwater Fish Carassius Auratus Research Articles

Biotransformation of dietary inorganic arsenic in a freshwater fish Carassius auratus and the unique association between arsenic dimethylation and oxidative damage

The metabolic process and toxicity mechanism of dietary inorganic arsenic (iAs) in freshwater fish remain unclear to date. The present study conducted two iAs [arsenate (As(V)) and arsenite (As(III))] dietary exposures in freshwater fish crucian carp (Carassius auratus). The fish were fed on As supplemented artificial diets at nominal concentrations of 50 and 100 μg As(III) or As(V) g−1 (dry weight) for 10 d and 20 d. We found that the liver, kidney, spleen, and intestine of fish accumulated more As in As(V) feeding group than that in As(III), while the total As levels in muscle were similar between As(V) and As(III) group at the end of exposure. Reduction of As(V) to As(III) and oxidation of As(III) to As(V) occurred in fish fed with As(V) and As(III), respectively, indicating that toxicity of iAs was likely elevated or reduced when iAs was absorbed by fish before entering into human body through diet. Biomethylation to monomethylarsonic acid and dimethylarsinic acid and transformation to arsenocholine and arsenobetaine were also found in the fish. The linear regression analysis showed a positive correlation between secondary methylation index and the malondialdehyde content in tissues, highlighting the vital role of arsenic dimethylation in the oxidative damages in fish.

Responses of antioxidant defense system to polyfluorinated dibenzo-p-dioxins (PFDDs) exposure in liver of freshwater fish Carassius auratus

In this study, we evaluated the toxicity of ten polyfluorinated dibenzo-p-dioxins (PFDDs) congeners to freshwater fish Carassius auratus, by determining the antioxidative responses and lipid peroxidation in the liver after the fish were injected with two different concentrations (10 and 100µmol/kg) of individual PFDDs for 3 and 14 days. The results showed that oxidative stress was obviously induced in some PFDDs-treated groups, as implied by the significantly inhibited antioxidants levels (superoxide dismutase, catalase, reduced glutathione, and glutathione S-transferase) and elevated malondialdehyde content. In addition, the oxidative stress inducing ability was variable for different PFDDs congeners, which was related with the substitution number and position of fluorine atom. Based on the calculated integrated biomarker response (IBR) values, the toxicity was ranked as 2,3,7,8-FDD>Octa-FDD>1,2,3,4,7-FDD>1,3,6,8-FDD>1,2,3,4,6,7-FDD>1,2,6,7-FDD>1,2,7-FDD>DD>2,7-FDD>2-FDD. This study can enhance the general understanding of the PFDDs induced oxidative stress in aquatic organisms.

Hepatic oxidative stress biomarker responses in freshwater fish Carassius auratus exposed to four benzophenone UV filters

Benzophenone (BP) type UV filters are widely used in many personal care products to protect human from UV irradiation. Despite the estrogenic potencies to fish and the environmental occurrences of BP derivatives in aquatic systems, little information is available regarding their effects on the antioxidant defense system in fish. In this work, the oxidative stress induced in livers of Carassius auratus was assessed using four biomarkers. The integrated biomarker response (IBR) was applied to assess the overall antioxidant status in fish. Moreover, liver tissues were also studied histologically. The changes in the activities of antioxidant enzymes and glutathione levels suggested that BPs generates oxidative stress in fish. The IBR index revealed that the hepatic oxidative toxicity followed the order BP-1>BP-2>BP-4>BP-3. The histopathological analysis revealed lesions caused by BPs. This investigation provides essential information for assessing the potential ecological risk of BP-type UV filters.

Behavioral and biochemical responses in freshwater fish Carassius auratus exposed to sertraline

Sertraline is one of the most commonly prescribed selective serotonin reuptake inhibitors and is frequently detected in the aquatic environment. However, knowledge regarding relationships among molecular or biochemical endpoints involved in modes of action (MOAs) of sertraline and ecologically important behavioral responses of fish is insufficient. The present study aimed to investigate the bioconcentration and possible adverse outcomes pathways (AOPs) in crucian carp (Carassius auratus) exposed to various concentrations of sertraline (4.36, 21.3 and 116μgL−1) for 7d. Bioconcentration factor values were in the range of 19.5–626 in liver, 6.94–285 in brain, 4.01–146 in gill and 0.625–43.1 in muscle during the entire period of exposure. Liver superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities and brain acetylcholinesterase (AChE) activity were selected as biochemical endpoints associated with MOAs. Swimming activity, shoaling, feeding rate and food consumption were determined to assess behavioral responses. Fish plasma levels of sertraline exceeding human therapeutic doses were also predicted from external exposure concentrations. Significant enhancements in CAT, GPx, AChE and swimming activities and decreases in shoaling tendency, feeding rate and food consumption were observed when fish plasma levels exceeded human therapeutic thresholds. Shoaling, feeding rate and food consumption were correlated with the activities of SOD, CAT and GST. A significant positive correlation between swimming activity and AChE activity was also observed. As such, our study provides important AOPs linking biochemical responses with ultimate ecologically relevant behavioral endpoints.

Antioxidant status and Na(+), K (+)-ATPase activity in freshwater fish Carassius auratus exposed to different combustion products of Nafion 117 membrane: an integrated biomarker approach.

Nafion 117 membrane (N117), an important polymer electrolyte membrane (PEM), has been widely applied in numerous chemical technologies. Its increasing production and utilization will inevitably lead to the problem of waste disposal, with incineration as an important method. However, toxicity data of its combustion products on aquatic organisms have been seldom reported. The present study was therefore conducted to investigate the antioxidant response and Na(+), K(+)-ATPase activity in liver of Carassius auratus exposed to different combustion products of N117 for 5, 15, and 30 days. The concentrations of fluorine ion (F(-)) in the aquaria among the exposure durations were analyzed using the ion chromatography system. The results showed that these treatments have the capability to induce oxidative stress and suppress Na(+), K(+)-ATPase activity, as indicated by some significant alterations on these measured toxicity end-points in fish liver. According to the integrated biomarker response (IBR) index, the toxicity intensity of these experimental treatments was tentatively ranked. Taken together, these observations provided some preliminary data on the potential toxicity of the combustion products of N117 on aquatic organisms and could fill the information gaps in the toxicity database of the current-use PEM.

Bioaccumulation, distribution and metabolism of BDE-153 in the freshwater fish Carassius auratus after dietary exposure

Polybrominated diphenyl ethers (PBDEs) are of great environmental concern due to bioaccumulation and biomagnification in different food chains. However, significant biotransformation of some congeners via reductive debromination has been observed during in vivo and in vitro laboratory exposures. Little is known about the fate of 2,2′,4,4′,5,5′-hexabromodiphenyl ether (BDE-153) in fish. In the present study, crucian carp (Carassius auratus) were exposed to BDE-153 at a concentration of 10μg/g in food for 28 days. BDE-153 and its metabolites in different tissues were identified and quantified using gas chromatography coupled with tandem mass spectrometry and ultra-high performance liquid chromatography coupled with tandem mass spectrometry. In addition to eight debrominated metabolites, four oxidative metabolites were detected 4′-hydroxy-2,2′,4,5′-tetra-BDE, 6-hydroxy-2,2′,4,4′-tetra-BDE, 2,4-dibromophenol and 2,4,6-tribromophenol. With regard to the concentrations of BDE-153 and the major metabolites, the contribution order of different tissues was bile>brain>liver>gill>muscle. The highest concentrations of BDE-153 and metabolite 2,2′,4,4′-tetrabromodiphenyl ether were detected in bile at 808ng/g and 157ng/g, respectively. Our results suggested that three possible metabolic pathways of BDE-153 occurred in crucian carp via dietary exposure: debromination, hydroxylation and cleavage of the diphenyl ether bond. These findings indicated evidence of the bioavailability of BDE-153 from food in the form of debrominated metabolites and oxidative metabolites in freshwater fish, which is critical to understanding the complete risks associated with PBDE bioaccumulation and metabolism in humans and wildlife.

Metal accumulation and antioxidant defenses in the freshwater fish Carassius auratus in response to single and combined exposure to cadmium and hydroxylated multi-walled carbon nanotubes

The effects of cadmium, hydroxylated multi-walled carbon nanotubes, and their mixture on metal accumulation and antioxidant defenses were studied using the goldfish Carassius auratus as the test organism. The fish were exposed to 0.1mg/L Cd, 0.5mg/L OH-MWCNTs, or 0.1mg/L Cd+0.5mg/L OH-MWCNTs for 3 and 12 days. Then, the Cd concentration was determined in the gill, liver and muscle. Moreover, hepatic antioxidant enzyme activity (superoxide dismutase, catalase and glutathione peroxidase), glutathione level and malondialdehyde content were also measured. A continuous accumulation of Cd was observed throughout the experimental period. Cd accumulation in tissues occurred in the following order: gill>liver>muscle at 3 days and liver>gill>muscle at 12 days. The concentrations of Cd in the livers of fish exposed to the combination of Cd+OH-MWCNTs were significantly higher than those in fish exposed to either single chemical after 12 d of exposure. Meanwhile, the mixture evoked severe oxidative stress in the exposed fish, as indicated by significant inhibition of SOD, CAT and GPx activity, a remarkable decrease in GSH level, and simultaneous elevation of MDA content. These results suggested that the effect of the combined factors on metal accumulation and oxidative stress biomarkers was more obvious than that of single factors at longer exposure durations.

Movement patterns of three freshwater fish species after upstream transportation by fishway in the Jangheung Dam

We investigated the post-passage movement patterns of three freshwater fish species, Carassius auratus, Hemibarbus longirostris, and Silurus asotus, frequently captured in a fishway trap as they were released into the lentic dam reservoir. Acoustic telemetry was used to monitor fish movement. Acoustic tags were attached to 10 individuals (C. auratus, [n = 6], Ca1–6; H. longirostris, [n = 2], Hl1 and Hl2; and S. asotus, [n = 2], Sa1 and Sa2). The individual (Sa1) was particularly monitored to changes of vertical distribution between day and night a depth transmitter. The lentic species, C. auratus and S. asotus, were grouped into those that moved into upper streams (Ca1, Ca2, Ca3, and Sa1) and those that stayed in the reservoir (Ca4, Ca5, Ca6, and Sa2); all individuals utilized almost the entire area of the reservoir. However, H. longirostris, which favor lotic environments, showed different responses; one (Hl1) immediately moved into one of upper the streams, whereas the other (Hl2) utilized the entire dam reservoir area. No significant correlations were observed between size of the C. auratus individuals (total length and total weight) and monitored parameters (total movement distance, movement distance per day, and number of receivers encountered). Moreover, S. asotus used the water at a mean depth of 2.89 ± 1.52 m, and its vertical distribution changed more actively during the night than that during the day. Therefore, the efficacy of the Jangheung Dam fishway could be improved through release of the confluence of the inflow streams at the dam reservoir, which would provide both lentic and lotic environments without extended migration for fish.

Metal accumulation and oxidative stress biomarkers in liver of freshwater fish Carassius auratus following in vivo exposure to waterborne zinc under different pH values

In this study, laboratory experiments were conducted to investigate the combined effect of zinc and pH on metal accumulation and oxidative stress biomarkers in Carassius auratus. Fish were exposed to 0.1 and 1.0mg Zn/L at three pH values (5.0, 7.25, 9.0) for 3, 12, and 30d. After each exposure, the contents of three trace elements (Zn, Fe and Cu) were determined in liver. Generally, longer exposure to zinc (12d and 30d) increased hepatic Zn and Cu deposition, but decreased Fe content. Increasing accumulation of Zn in the tissue was also observed with increasing zinc concentration in the exposure medium. Moreover, hepatic antioxidant enzyme activities including superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), together with the level of glutathione (GSH) were measured to evaluate the oxidative stress status. The decreases in the four measured biochemical parameters after 3d exposure might reflect the failure of the antioxidant defense system in neutralizing the ROS generated during the metabolic process, while the recovery of the antioxidants at days 12 and 30 suggested a possible shift toward a detoxification mechanism. With regard to the influence of pH on zinc toxicity, the general observation was that the living environment became more stressful when the water conditions changed from an acidic state toward a near-neutral or alkaline state.

Comparative antioxidant status in freshwater fish Carassius auratus exposed to eight imidazolium bromide ionic liquids: A combined experimental and theoretical study

Imidazolium bromide ionic liquids such as 1-alkyl-3-methylimidazolium bromides ([AMIm]Br) and 1-alkyl-2,3-dimethylimidazolium bromides ([AMMIm]Br) are common-use organic salts. However, data on comparative toxicological effects of these ILs are lacking for fish. In this study, a combined experimental and theoretical approach was applied to compare and analyze the effects of these ILs on biochemical biomarkers in liver of Carassius auratus treated with different concentrations (2 and 20mg/L) for 3 and 16d. Changes in the activities of superoxide dismutase, catalase, glutathione peroxidase, and in the levels of reduced glutathione and malondialdehyde were detected, indicating that these ILs exhibit potential biotoxicity. The integrated biomarker response (IBR) index suggested that 1-hexyl-3-methylimidazolium bromide ([HMIm]Br), 1-octyl-3-methylimidazolium bromide ([OMIm]Br), 1-hexyl-2,3-dimethylimidazolium bromide ([HMMIm]Br), and 1-octyl-2,3-dimethylimidazolium bromide ([OMMIm]Br) showed the highest biotoxicity under different concentrations or exposure time, while 1-ethyl-3-methylimidazolium bromide ([EMIm]Br) always showed the least stressful power towards the test organism. Quantum chemical calculations (electronic parameters, frontier molecular orbitals, and Wiberg bond order) were also conducted to interpret the experimental results. Notably, some descriptors were correlated with the toxicity order. In addition, theoretical calculations provided some valuable information on metabolic pathways of these ILs, which may help to get better understanding on their environmental behavior and fate. In general, the toxicological determination and analysis of these ILs were performed with a combined experimental and theoretical method, which may contribute to the future ecotoxicological studies.

Bioaccumulation, inter-organ distribution, and retention of waterborne and dietary 241Am in silver crucian carp

The bioaccumulation of actinide 241Am from food and water in bodies of freshwater fish (Carassius auratus gibelio, silver crucian carp) has been investigated in laboratory experiments. Homogenized biomass of submerged macrophytes labeled with 241Am was injected into the fish gullet. Internal organs and tissues of crucian carp could be ranked according to activity concentration of dietary 241Am after depuration of the digestive tract as follows: liver > gonads > bones > muscles. Accumulation of waterborne 241Am in internal organs and tissues of crucian carp mainly occurred via the digestive tract. While the concentration of 241Am in liver of crucian carps decreased during depuration time, its concentration in bones increased indicating slower transfer of 241Am to skeleton compared to muscles and liver. The retention of dietary 241Am in the bodies of crucian carp reached 35%–46% of ingested 241Am: 20%–31% was retained in liver, 0.6%–0.8% in skeleton, and 1.4%–2.0% in muscles. The concentration factor of 241Am from water was 0.4 for the whole body, 0.3 for liver, 0.01 for muscles, and 0.01 for skeleton. Trace amounts of 241Am were recorded in viscera and muscles of a wild population of silver crucian carp inhabiting a radioactively contaminated part of the Yenisei River.

Effect of Sub-Acute Exposure to Nickel on Hematological and Biochemical Indices in Gold Fish (Carassius auratus)

The increasing use of heavy metals in industry in the modern world unfavorably affects the aquatic environment. Acute toxicity of nickel to the fresh water fish carassius auratus was determined using Probite analyze, Fish were exposed to selected concentration of nickel and the mortality data were determined after 24, 48, 72 and 96 hours. LC50 values for 24,48,72 and 96 hours were calculated with the 95% fiducially limits. The 24,48,72 and 96 hours LC50 values of nickel to the fish were 161.78 ± 3.05, 130.58 ± 2.32, 110.19 ± 1.57, 100.39 ± 0.54 ppm respectively. White blood cell (WBC) count, hemoglobin (Hb) and hematocrit (Hct) level were significantly reduced at experimental concentrations (p 0.05). In summary, nickel intoxication resulted indicated that hematological and biochemical parameters can be used as an indicator of nickel related stress in fish on exposure to elevated nickel status.

Bioconcentration, metabolism, and biomarker responses in freshwater fish Carassius auratus exposed to roxithromycin

To investigate the distribution, bioconcentration, metabolism, and biomarker responses of macrolide antibiotic roxithromycin (ROX) in fish, crucian carp (Carassius auratus) were exposed to various concentrations of ROX (4, 20, and 100μgL−1) for 20d. The ROX content in different tissues was quantified using UPLC/MS/MS. The liver exhibited the highest ROX concentration followed by the bile, gills, and muscle tissues. After 15d of exposure to different concentrations of ROX, the bioconcentration factors were 2.15–38.0 in the liver, 0.950–20.7 in the bile, 0.0506–19.7 in the gill, and 0.0439–13.8 in the muscle; these results were comparable to the estimated BCF values. The metabolites formed in the bile were identified based on metabolic identification in human bile. Additionally, the biomarkers, including acetylcholinesterase in the brain, as well as 7-ethoxyresorufin O-deethylase and superoxide dismutase in the liver changed significantly after 5, 10, 15, and 20d of exposure (P<0.05). Our results suggest that ROX can accumulate and be metabolized in fish; therefore, interactions between ROX or its metabolites and the biological systems may induce biochemical disturbances in fish.

Comparative antioxidant status in freshwater fish Carassius auratus exposed to six current-use brominated flame retardants: A combined experimental and theoretical study

Decabromodiphenyl ether (BDE-209) and several non-polybrominated diphenyl ether (PBDE) brominated flame retardants (BFRs), such as tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD), decabromodiphenyl ethane (DBDPE), hexabromobenzene (HBB) and pentabromotoluene (PBT), are persistent halogenated contaminants ubiquitously detected in aquatic systems. However, data on comparative toxicological effects of these BFRs are lacking for fish. In this study, a combined experimental and theoretical approach was used to compare and analyze the effects of these BFRs on biochemical biomarkers in liver of Carassius auratus injected intraperitoneally with different doses (10 and 100mg/kg) for 7, 14 and 30 days. Oxidative stress was evoked evidently for the prolonged exposure, represented by the significantly altered indices (superoxide dismutase, catalase, glutathione peroxidase, reduced glutathione, and malondialdehyde). The integrated biomarker response (IBR) index ranked biotoxicity as: PBT>HBB>HBCD>TBBPA>BDE-209>DBDPE. Quantum chemical calculations (electronic parameters, frontier molecular orbitals, and Wiberg bond order) were performed for theoretical analysis. Notably, some descriptors were correlated with the toxicity order, probably implying the existence of a potential structure–activity relationship when more BFRs were included. Besides, theoretical calculations also provided some valuable information regarding the molecular characteristics and metabolic pathways of these current-use BFRs, which may facilitate the understanding on their environmental behavior and fate. Overall, this study adopted a combined experimental and theoretical method for the toxicological determination and analysis of the BFRs, which may also be considered in future ecotoxicological studies.

Oxidative stress biomarkers in freshwater fish Carassius auratus exposed to decabromodiphenyl ether and ethane, or their mixture

Decabromodiphenyl ether (BDE-209) and its commercial alternative decabromodiphenyl ethane (DBDPE) are two structurally similar brominated flame retardants, with evidence of their ubiquitous existence in aquatic ecosystems. The present study was conducted to investigate the hepatic oxidative stress inducing potential of BDE-209, DBDPE, and their mixture in Carassius auratus after exposure to different doses (10, 50 and 100 mg/kg) for 7, 14 and 30 days. Results showed that oxidative stress was evoked evidently for the experimental groups with longer exposure duration, as indicated by significant inhibition in the antioxidant enzymes activities and decrease in the reduced glutathione level, as well as simultaneous elevation of lipid peroxidation level measured by malondialdehyde content. In addition, it was found that BDE-209 possessed a higher oxidative stress inducing ability than DBDPE. Considering the more pronounced antioxidant responses in combined exposure, the interaction of BDE-209 and DBDPE was presumed to be additive action.

Cytotoxicity of cuprous oxide nanoparticles to fish blood cells: hemolysis and internalization

Cuprous oxide nanoparticles (Cu2O NPs) possess unique physical and chemical properties which are employed in a broad variety of applications. However, little is known about the adverse effects of Cu2O NPs on organisms. In the current study, in vitro cytotoxicity of Cu2O NPs (ca. 60 nm in diameter) to the blood cells of freshwater fish Carassius auratus was evaluated. A concentration-dependent hemolytic activity of Cu2O NPs to red blood cells (RBCs) and the phagocytosis of Cu2O NPs by leukocytes were revealed. The results showed that dosages of Cu2O NPs greater than 40 μg/mL were toxic to blood cells, and could cause serious membrane damage to RBCs. The EC50 value of Cu2O NPs as obtained from RBCs and whole blood exposure was 26 and 63 μg/mL, respectively. The generation of reactive oxygen species and the direct interaction between Cu2O NPs and the cell membrane were suggested as the possible mechanism for cytotoxicity, and the intrinsic hemolytic active of Cu2O NPs was the main contributor to the toxicity rather than solubilized copper ions. The adsorption of plasma proteins on the surfaces of Cu2O NPs led to their aggregation in whole blood, and aggregate formation can significantly alleviate the hemolytic effect and subsequently mediate the phagocytosis of Cu2O NPs by leukocytes.

The Dose–Response Relationships for EROD and GST Induced by Polyaromatic Hydrocarbons in Carassius auratus

Freshwater fish Carassius auratus were chosen as experimental animals, the hepatic biochemical responses to medium-term exposure of five PAHs were measured as ethoxyresorufin O-deethylase (EROD) activity (phase I) and glutathione S-transferase (GST) activity (phase II) to assess sub-lethal effects. The fold increases of EROD and GST activity were calculated and both increased in the order Fluoranthene < Fluorene < Benzo(b)fluoranthene < Benzo(g,h,i)perylene < Indeno(1,2,3-cd)-pyrene. The clear dose-response relationships were found for liver EROD and GST activity induced by PAHs. The enzyme EROD and GST in Carassius auratus were confirmed as useful biomarkers of exposure to both PAH and PAH-like compounds.

Nationwide study of dioxins in the freshwater fish Carassius auratus ( gibelio) langsdorfii (crucian carp) in Japan: Concentrations and estimation of source contribution ratios

We investigated dioxin concentrations in freshwater fish in Japan by standardizing species to detect subtle decreasing trends of dioxin concentrations in the future with the reinforcement of regulations. The fish studied were crucian carp ( Carassius auratus ( gibelio) langsdorfii), an omnivorous species. Fish and sediments were collected from 14 rivers and lakes located in remote areas, agricultural areas, and small and large cities throughout Japan. The total toxic equivalent (TEQ) dioxin concentrations at the three remote sites were about 20% of the concentrations at the other 11 sites, which all had similar concentrations. The average concentrations in fish collected from these 11 sites were 0.69 pg TEQ g −1 wet wt. (95% CI 0.52–0.85) or 57.3 pg TEQ g −1 fat (95% CI 47.9–66.7). There were notable differences in congener profiles of polychlorinated p-dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) between sampling areas. A chemical mass balance method was used to estimate the proportions of dioxin contributions from different sources. The sampling areas were classified into four groups on the basis of the predominant source of dioxin contamination: a combustion group, a chlornitrofen herbicide group, a chlornitrofen and pentachlorphenol herbicide group, and a remote group. The congener profiles of dioxin like polychlorinated biphenyls (DL-PCBs) were almost the same in all areas and close to those in Kanechlor, which indicates that pollution from PCB products has spread throughout Japan. From samples collected during the spawning season, it was determined that about 20% of the dioxins in adult females were transferred to the eggs, resulting in higher dioxin concentrations in adult males than in females. Biota–sediment accumulation factors (BSAFs) of 2,3,7,8-substituted PCDDs/DFs for crucian carp were larger than those of non-2,3,7,8-substituted congeners, and BASFs decreased with increasing number of chlorines. The BSAFs of DL-PCBs were 10 times greater than those of PCDDs/DFs, and BSAFs of mono- ortho type DL-PCBs were higher than those of non- ortho types.

Bioaccumulation, depuration and oxidative stress in fish Carassius auratus under phenanthrene exposure

In this study, laboratory experiment was carried out to determine phenanthrene bioaccumulation, depuration in whole fish and oxidative stress in the liver of freshwater fish Carassius auratus. Fish were exposed to 0.05 mg/l phenanthrene for different periods, while one control group was designated for each exposure group. Some fish after 7 days of exposure were transferred to diluted water. The concentrations of phenanthrene in fish were analyzed by HPLC. Twenty-four hours after the exposure, reactive oxygen species (ROS) were trapped by phenyl- tert-butylnitrone and detected by electron paramagnetic resonance (EPR). The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione-s-transferase (GST) were also determined. The concentrations of phenanthrene in fish increased rapidly shortly after the start of the exposure, reached a maximum level at the 2 days, and then it declined quickly to low-level-steady state. The elimination process of phenanthrene could be divided into two periods—a fast elimination period following a slower loss period. The elimination curve could be fitted mathematically as the sum of two exponential functions according to two-compartment model: C( t) = 2.72e −1.065 t + 0.68e −0.0364 t . The PBN-radical adducts were detected in fish liver samples following the exposure 24 h. The hyperfine splitting constants for the PBN-radical adducts were a N = 13.5 G, a H = 1.77 G and g value was 2.0058, which were consistent with those of PBN/ OH. The results indicated that the hydroxyl radical was probably significantly induced during the exposure of phenanthrene, as compared to the control group. The changes of activities of the antioxidant enzymes also were observed. In addition, after fish were removed from phenanthrene exposure, the recovery status of these antioxidant indices was explored. These results clearly indicated phenanthrene could be accumulated in fish and similar redox cyclings were produced, resulting in the changes of the activities of the antioxidant enzymes and the production of ROS with the oxidative stress.

Electron paramagnetic resonance evidence of hydroxyl radical generation and oxidative damage induced by tetrabromobisphenol A in Carassius auratus

Tetrabromobisphenol A (TBBPA) is one of the most widely used brominated flame retardants (BFRs). To confirm its putative oxidative stress-inducing activity, freshwater fish Carassius auratus were injected intraperitoneally with TBBPA. One experiment lasted 3 h to 28 days after a single injection of 100 mg/kg TBBPA, and the other lasted 24 h after a single injection of 0–300 mg/kg TBBPA. Reactive oxygen species (ROS) were trapped by phenyl- tert-butyl nitrone (PBN) and detected by electron paramagnetic resonance (EPR). Protein carbonyl (PCO) and lipid peroxidation product (LPO) content were also determined. A six-line EPR spectrum was detected in the sample prepared in air, and a multiple one was obtained in nitrogen. The observed spectrum in nitrogen fits the simulation one with PBN/ OCH 3 and PBN/ CH 3 quite well. As compared to the control group, TBBPA significantly induced ROS production marked by the intensity of the prominent spectra in liver and bile. TBBPA (100 mg/kg) also significantly increased PCO content in liver starting 24 h and LPO content 3 days after injection. Either PCO or LPO content showed significant relation with ROS production. Based on the hyperfine constants and shape of the spectrum, ROS induced by TBBPA was determined as OH. The results clearly indicated that TBBPA could induce OH generation and result in oxidative damage in liver of C. auratus.