Liver Of Crucian Carp Research Articles

Immunometabolic interplay in Edwardsiella tarda-infected crucian carp (Carassius auratus) and in vitro identification of the antimicrobial activity of apolipoprotein D (ApoD) by utilization of multiomics analyses

Edwardsiella tarda is an intracellular pathogenic bacteria that can imperil the health of farmed fish. However, the interactive networks of immune regulation and metabolic response in E. tarda-infected fish are still unclear. In this investigation, we aimed to explore immunometabolic interplay in crucian carp after E. tarda infection by utilizing multiomics analyses. Crucian carp (Carassius auratus) receiving E. tarda infection showed increased levels of tissue damage and oxidative injury in liver. Multiomics analyses suggested that carbon and amino acid metabolism may be considered as crucial metabolic pathways in liver of crucian carp following E. tarda infection, while spaglumic acid, isocitric acid and tetrahydrocortisone were the crucial liver biomarkers. After that, a potential antimicrobial peptide (AMP) sequence called apolipoprotein D (ApoD) was identified from omics study. Then, tissue-specific analysis indicated that liver CaApoD showed the highest expression among isolated tissues. After Aeromonas hydrophila stimulated, CaApoD expressions increased sharply in immune-related tissues. Moreover, CaApoD fusion protein could mediate the in vitro binding to A. hydrophila and E. tarda, attenuate bacterial growth as well as diminish bacterial biofilm forming activity. These findings may have a comprehensive implication for understanding immunometabolic response in crucian carp upon infection.

Enrichment and distribution of 3,6-dichlorocarbazole in red crucian carp (Carassius auratus) and its hepatotoxicity

Polyhalogenated carbazoles (PHCZs) are a class of organohalogen compounds where the hydrogen atom on the carbazole ring is replaced by a halogen atom. In recent years, PHCZs have drawn increasing concern due to their persistence, dioxin-like toxicity, bioaccumulation, potential ecological hazards and widespread occurrence in the environment. Current research on the enrichment and depuration of PHCZs in biological tissues and organs is insufficient, and the liver toxicity is unclear. Herein, to understand the enrichment and elimination of 3,6-DCCZ in fish tissues and organs as well as the hepatotoxicity, we exposed the red crucian carp to 20 and 100 μg/L of 3,6-DCCZ for 20 days followed by a depuration period of 10 days. The 3,6-DCCZ enrichment in each organ tissue was classified from high to low: brain > liver, intestine, gill > muscle. For depuration, 3,6-DCCZ was quickly excreted in the various organs of the red crucian carp; however, the liver depuration was slow, with the concentration of 3,6-DCCZ was maintained at 0.25–0.35 μg/g. 3,6-DCCZ exposure at both tested concentrations induced oxidative stress in red crucian carp, causing lipid peroxidation and DNA damage, as well as some histopathological changes in the liver, such as cell vacuolization, nucleus pyknosis, nucleus pleomorphism, no nucleus areas. Additionally, the 3,6-DCCZ exposure at higher concentration (100 μg/L) caused more serious damage and abnormal lipid metabolism in the red crucian carp liver.

Stress response and tolerance mechanisms of NaHCO3 exposure based on biochemical assays and multi-omics approach in the liver of crucian carp (Carassius auratus)

The development and utilization of saline-alkaline water, an important backup resource, has received widespread attention. However, the underuse of saline-alkaline water, threatened by the single species of saline-alkaline aquaculture, seriously affects the development of the fishery economy. In this work, a 30-day NaHCO3 stress experimental study combined with analyses of untargeted metabolomics, transcriptome, and biochemical approaches was conducted on crucian carp to provide a better understanding of the saline-alkaline stress response mechanism in freshwater fish. This work revealed the relationships among the biochemical parameters, endogenous differentially expressed metabolites (DEMs), and differentially expressed genes (DEGs) in the crucian carp livers. The biochemical analysis showed that NaHCO3 exposure changed the levels of several physiological parameters associated with the liver, including antioxidant enzymes (SOD, CAT, GSH-Px), MDA, AKP, and CPS. According to the metabolomics study, 90 DEMs are involved in various metabolic pathways such as ketone synthesis and degradation metabolism, glycerophospholipid metabolism, arachidonic acid metabolism, and linoleic acid metabolism. In addition, transcriptomics data analysis showed that a total of 301 DEGs were screened between the control group and the high NaHCO3 concentration group, of which 129 up-regulated genes and 172 down-regulated genes. Overall, NaHCO3 exposure could cause lipid metabolism disorders and induce energy metabolism imbalance in the crucian carp liver. Simultaneously, crucian carp might regulate its saline-alkaline resistance mechanism by enhancing the synthesis of glycerophospholipid metabolism, ketone bodies, and degradation metabolism, at the same time increasing the vitality of antioxidant enzymes (SOD, CAT, GSH-Px) and nonspecific immune enzyme (AKP). Herein, all results will provide new insights into the molecular mechanisms underlying the stress responses and tolerance to saline-alkaline exposure in crucian carp.

Zinc oxide nanoparticles (ZnO-NPs) exhibit immune toxicity to crucian carp (Carassius carassius) by neutrophil extracellular traps (NETs) release and oxidative stress

Zinc oxide nanoparticles (ZnO-NPs) are widely used in sunscreens, cosmetics, paint, construction materials, and other products. ZnO-NPs released into the environment can harm aquatic creatures and pose a health risk to humans through the food chain. ZnO-NPs are toxic to fish, but there are few reports on its immunotoxicity on crucian carp (Carassius carassius). In this study, ZnO-NPs increased the biochemical indexes of the liver in serum, including aspartate aminotransferase (AST) and alanine aminotransferase (ALT). In histopathological observation, many inflammatory cells were filled in the liver's central vein stimulated by ZnO-NPs. Furthermore, ZnO-NPs could increase malondialdehyde (MDA) level, lessen superoxide dismutase (SOD) level, and elevate the level of neutrophil extracellular traps (NETs). However, deoxyribonuclease I (DNase I) alleviated all biochemical indexes and histopathological changes. Immunofluorescence in vitro confirmed that NETs were composed of citrullinated histone 3, myeloperoxidase, and neutrophil elastase. ZnO-NPs-increased NETs were dependent on reactive oxygen species (ROS) and nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase and were also related to partial processes of glycolysis. Our study confirms that ZnO-NPS has a toxic effect on the liver of crucian carp. DNase I can prevent liver damage caused by ZnO-NPs, which provides a new insight into the immunotoxicity of ZnO-NPs to fish.

Comparison of physicochemical properties of different tissues from China climbing perch Anabas testudineus and crucian carp Carassius auratus.

This study aimed to investigate nutrition in climbing perch Anabas testudineus which is an important nutritious economic freshwater fish in Asia and compare with Carassius auratus (crucian carp). Three kinds of tissues, including muscle, livers, and eggs, were isolated, respectively. Physicochemical properties including moisture, ash, protein, amino acids, fat, vitamins, and calcium contents in those tissues were determined. The results showed climbing perch muscle and liver contained less moisture, but more protein, amino acids, and vitamins than crucian carp muscle and liver. While moisture, ash, protein, and total amino acids contents of climbing perch egg were lower than those of crucian carp egg. Climbing perch egg had more fat, vitamins, and calcium than crucian carp egg. The amino acid profile was also performed, and 16 amino acids were identified and quantified in muscle, liver, and egg. Among tissues, the highest and lowest concentration of total amino acid content was found in crucian carp eggs and livers, respectively. Glutamic acid (Glu) and histidine (His) were the most and least amino acids in climbing perch and crucian carp tissues, respectively. Sixteen amino acids in climbing perch egg were less than those in crucian carp egg while it is an opposite case in muscle and liver, which amino acids of climbing perch tissues were more than those of crucian carp muscle and liver. Vitamin A of climbing perch was more than crucian carp in all three tissues, but vitamin E content in climbing perch liver was lower than that of crucian carp liver. Calcium content of muscle had no difference between two species. The abovementioned comparison of physicochemical properties of different tissues from China climbing perch and crucian carp will provide a necessary supplementary of freshwater muscle nutrition research, also was helpful for application of climbing perch.

Toxicity assessment of municipal sewage treatment plant effluent by an integrated biomarker response in the liver of crucian carp (Carassius auratus).

In this study, crucian carp (Carassius auratus) was exposed to the increasing concentrations of municipal sewage treatment plant effluent (MSTPE) for 15days, and the activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and acetylcholinesterase (AChE), together with the contents of malondialdehyde (MDA) and glutathione (GSH) in the liver of C. auratus were investigated. Moreover, the integrated biomarker response (IBR) approach was applied to assess the adverse effects of MSTPE in freshwater. The aim of the study was to provide an effective biological indicator for evaluating the toxicity effects and ecological risks of MSTPE in the freshwater environment quantitatively. Results showed that MSTPE could cause oxidative damage to the liver of C. auratus, which reflected through the increasing MDA content over the exposure period. MSTPE also led to the biochemical responses of antioxidant defense in C. auratus liver, such as the enhancement of SOD, CAT, and GPx activities, as well as the inhibition of AChE activity and GSH content. It was found that MDA, SOD, GPx, and GSH could be used as the biomarkers for reflecting the adverse effects of MSTPE in the receiving freshwater on the 12th day of exposure. A significant increase of IBR values was observed as the increasing concentration of MSTPE, and the IBR values presented a significant positive correlation (r = 0.891, P < 0.05) with the increasing concentrations of MSTPE, indicating that IBR approach is a promising tool for assessing the toxicity effects of MSTPE in environmental freshwater.

In vivo effects of neomycin sulfate on non-specific immunity, oxidative damage and replication of cyprinid herpesvirus 2 in crucian carp (Carassius auratus gibelio)

Neomycin belongs to the family of 2-deoxystreptamine-containing aminoglycoside antibiotics. It is widely used for bacterial infections, targeting most gram-negative and some gram-positive bacteria. Neomycin has also been reported to show antiviral activity. Here, we evaluated the toxicity of neomycin sulfate, and investigated its effect on non-specific immunity and viral infection in crucian carp (Carassius auratus gibelio). The safe concentration of neomycin sulfate for crucian carp was determined to be 102.9 mg/kg in vivo. In oxidative damage assays, neomycin sulfate increased superoxide dismutase and catalase activity and decreased malondialdehyde in the liver of crucian carp. In non-specific blood immune assays, the white blood cell count and complement 3 content significantly increased after neomycin sulfate treatment, while no significant changes were observed in antibacterial or lysozyme activity. In a challenge test, neomycin sulfate protected the crucian carp from cyprinid herpesvirus 2 (CyHV-2) infection and inhibited CyHV-2 replication. In cytotoxicity assays, low concentrations of neomycin sulfate had no cytotoxicity on cells from the fins of crucian carp. The results of the present study indicate that oral administration of neomycin sulfate reduced oxidative damage, enhanced immunity and provided protection against CyHV-2 in crucian carp.

The occurrence and distribution of hexabromocyclododecanes in freshwater systems, focusing on tissue-specific bioaccumulation in crucian carp

The occurrence and distribution of hexabromocyclododecanes (HBCDs) were investigated in freshwater, sediment, and selected crucian carp (Carassius carassius) tissues (muscle, liver, egg, and blood) to evaluate the potential for HBCDs bioaccumulation. The HBCDs concentration ranged from not detected to 0.35ng/L in freshwater, and from 0.037 to 35.4ng/g-dw in sediment. The highest HBCDs concentration was detected in crucian carp liver (5.14±8.15ng/g-ww), followed by egg (3.88±10.1ng/g-ww), blood (0.61±0.63ng/mL), and muscle (0.38±0.70ng/g-ww). In all crucian carp tissues, α-HBCD was the predominant stereoisomer, and the fraction of α-HBCD as a proportion of the total HBCDs in liver tissue (96%) was higher than that in egg tissue (79%). There was a positive correlation between the HBCDs concentration in crucian carp muscle and body size (p<0.01, Spearman). The biota–sediment accumulation factor (BSAF) (0.14) and bioconcentration factor (BCF) (137,000L/kg) values were estimated in crucian carp muscle using field-based data.

Complex responses to Si quantum dots accumulation in carp liver tissue: Beyond oxidative stress

The use of quantum dots (QDs) in biomedical applications is limited due to their inherent toxicity caused by the heavy metal core of the particles. Consequently, silicon-based QDs are expected to display diminished toxicity. We investigated the in vivo effects induced by Si/SiO2 QDs intraperitoneally injected in crucian carp liver. The QDs contained a crystalline Si core encased in a SiO2 shell, with a size between 2.75 and 11.25nm and possess intrinsic fluorescence (Ex 325nm/Em ∼690nm). Tissue fluorescence microscopy analysis revealed the presence of QDs in the liver for at least 2weeks after injection. Although protein and lipid oxidative stress markers showed the onset of oxidative stress, the hepatic tissue exhibited significant antioxidant adaptations (increase of antioxidant enzymes, recovery of glutathione levels), sustained by the activation of Hsp30 and Hsp70 chaperoning proteins. The increased activity of cyclooxigenase-2 (COX-2) and matrix metalloproteinases (MMPs) support the idea that Si/SiO2 QDs have a potential to induce inflammatory response, a scenario also indicated by the profile of Hsp60 and Hsp90 heat shock proteins. MMPs profile and the recovery of oxidative stress markers suggested a tissue remodelation phase after 3weeks from QDs administration.

Biological fate and effects of propranolol in an experimental aquatic food chain

The aim of this study was to evaluate the trophic transfer of the β-blocker propranolol (PRP) in an experimental aquatic food chain involving the green algae Scenedesmus obliquus, the water flea Daphnia magna and the crucian carp Carassius auratus, as well as the metabolism and effects of PRP in the liver of crucian carp. After a 48h PRP aqueous exposure for algae, with a subsequent 48h dietary exposure for daphnia and an 8d dietary exposure for crucian carp, PRP was observed in each trophic level, despite significant bioaccumulation did not occur in daphnia and crucian carp. A portion of the absorbed PRP was metabolized by the crucian carp to N-desisopropylated propranolol, propranolol glucuronic acid, monohydroxylated propranolol, hydroxypropranolol glucuronide and dihydroxypropranolol glucuronide, which were similar to those in mammals. In addition, multiple biomarkers in the liver of crucian carp (7-ethoxyresorufin O-deethylase, EROD; 7-benzyloxyresorufin O-dealkylation, BROD; superoxide dismutase, SOD and malondialdehyde, MDA) were measured. BROD and MDA were not significantly affected by PRP, while EROD and SOD did change significantly during the 8d dietary exposure. This work indicated that the trophic transfer of PRP, resulting in biochemical perturbations of fish biological systems, should be a concern for the assessment of the environmental risks to aquatic food chains.

Effects of a dietary administration of purple coneflower (Echinacea purpurea) on growth, antioxidant activities and 8 miRNAs expressions in crucian carp (Carassius auratus)

Echinacea purpurea (EP), a globally popular herbal medicine, has been used to treat various diseases in human and animals. However, little has been reported about its effects in fish. In this study, crucian carp (Carassius auratus) were selected to evaluate the effects of EP on growth performance and antioxidant response and the expressions of microRNAs. The results showed EP could stimulate the growth of crucian carp with the best effect was observed at dose of 4 g kg−1. In serum, the content of hydroxyl radical (·OH) and malondialdehyde (MDA) decreased by EP supplementation, whereas the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) increased. Similarly, the content of ·OH decreased, and the activities of CAT, GPX and GR increased in liver of crucian carp. Furthermore, in livers of crucian carp, EP supplements upregulated the expressions of the microRNAs (miR-16, miR-21, miR-125b, miR-146a, miR-155, miR-181a and miR-223), which had been confirmed to participate in regulating antioxidant and immune function in mammals. Our results suggest EP supplements in diets stimulated growth performance and antioxidant response of crucian carp. In liver, the upregulation of specific miRNAs may participate in the antioxidant effects of EP diets.

The bioaccumulation and biotransformation of synthetic estrogen quinestrol in crucian carp

The occurrence and fate of endocrine disrupting chemicals (EDCs) in aquatic species have attracted close attention during the last decades. In this study, the bioaccumulation and biotransformation of synthetic estrogen quinestrol, one of the typical EDCs, in the plasma and liver of crucian carp, were investigated by a newly developed and validated reversed-phase high performance liquid chromatography with fluorescent detection method. Crucian carp were exposed to quinestrol in concentration of 2, 10, 50, 100μg/L (5.49, 27.43, 137.17, 274.34nmol/L) for 60 days. After 60 days’ exposure, the concentrations of quinestrol found in liver and plasma were in the range of 0.25–0.69mg/kg and 0.19–0.30mg/L respectively, positively correlated with the exposure concentrations ranged 2–100μg/L (5.49–274.34nmol/L). There was a negative correlation between the bio-accumulation ratios and the exposure concentrations of quinestrol. 17α-Ethinylestradiol was also found in liver and plasma, and the concentrations were 0.02–0.19mg/kg and 0.37–0.96mg/L, respectively. The results indicated that quinestrol can be accumulated and transformed to 17α-ethinylestradiol in crucian carp. Moreover, exposure to quinestrol caused oxidative damages to crucian carp and the content of malondialdehyde increased in all treatment concentrations.

Multibiomarker responses upon exposure to tetrabromobisphenol A in the freshwater fish Carassius auratus

Tetrabromobisphenol A (TBBPA) is a widely used brominated flame retardant. It has been released into aquatic environments, where it is toxic to aquatic organisms. In the present study, five enzymes, glutathione-S-transferase (GST), aspartate aminotransferase (AST), alanine aminotransferase (ALT), UDP-glucuronosyltransferase (UDPGT), and the antioxidant enzyme glutathione reductase (GR) in serum and liver of crucian carp (Carassius auratus) were selected for screening. These enzymes may be suitable for use as early warning indicators of chronic TBBPA exposure. UDPGT, AST, ALT, and GR activities in serum were found to be as more sensitive to TBBPA as those of the liver. When the concentration of TBBPA exceeded 0.50–0.71mg/L for an exposure period of 32 days, GST, AST, ALT, and UDPGT activities cannot be restored to normal levels, suggesting that fish exposed to TBBPA above this threshold may incur irreversible damage. The activities of AST, ALT, and GR increased more significantly than GST and UDPGT at the lowest concentration of 0.35mg/L. AST showed the strongest activity with respect to toxic kinetics, followed by ALT and GR. This remained true from day 4 of exposure to TBBPA to day 32. However, GR showed the clearest and most significant dose–effect relationship. This shows that each of these three enzymes can be used as a biomarker for early warning applications focusing on TBBPA pollution. AST and ALT are suitable for use in conventional monitoring of water quality in areas at risk for TBBPA pollution, and GR is more suitable for use in burst TBBPA pollution accidents where GR activity in fish would change with the TBBPA concentration of the flowing water.

Identification of Metabolites and Pathway in Liver Microsomes of Phoxim in Crucian Carp by High Performance Liquid Chromatography and Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry

The liver microsomes were prepared by differential ultracentrifugation for the investigation of in vitro metabolism of phoxim.The metabolites were extracted twice from the reaction solution with ethyl acetate,remove the organic phase to rotary evaporator by near dryness at 40 ℃,then constant volume by 1mL methanol,separated and determined by HPLC,and then identified by HPLC-MS(ESI).Three major metabolites(M1: O,O,O’,O’-tetraethyldithiopyrophosphate,M2: diethylthiophosphate and M3: phoxom) indicated that phoxim underwent desulfur-oxidation and hydrolysis in crucian carp liver microsomes.Therefore,this study deduced the metabolic pathway and metabolic characteristic of phoxim in liver microsomes of crucian carp and laid a solid foundation for further study of its metabolism and metabolic enzymes.

Impairment of the mitochondrial oxidative phosphorylation system and oxidative stress in liver of crucian carp (Carassius auratus L.) exposed to microcystins

Hepatotoxic microcystins (MCs) are produced by cyanobacteria in diverse water bodies and the pathophysiology includes induction of reactive oxygen species and adenosine triphosphate (ATP) depletion in cells. In this study, we evaluated MCs induced changes in the oxidative phosphorylation (OXPHOS) system in mitochondria of crucian carp liver. Fish were subdivided into two groups that were intraperitoneally injected with two doses of MCs (50 and 200 MC-LReq μg/kg bw) and were sacrificed at 1, 3, 12, 24, and 48 h postinjection. The activities of five enzyme complexes of electron transport chain and mRNA expression of mitochondrial-encoded genes (cox1, cox2, cox3, and atp6) were significantly reduced in a time-dependent pattern after injection. There were also changes in mitochondrial ultrastructure, decreases in ATPase activities and reduction in antioxidant level after MCs exposure. Disorder in the OXPHOS system and decreased activities of antioxidative enzymes might contribute to bioenergy deficiency and consequent hepatocyte damage induced by MCs.

Inhibition of CYP450 1A and 3A by berberine in crucian carp Carassius auratus gibelio

Berberine has long been considered as an antibiotic candidate in aquaculture. However, studies regarding its effects on drug-metabolizing enzymes in fish are still limited. In the present study, the effects of berberine on cytochrome P4501A (CYP1A) and CYP3A in crucian carp were investigated. Injection of different concentrations of berberine (0, 5, 25, 50, and 100mg/kg) inhibited the CYP1A mRNA expression, thereby inhibiting further the catalytic activity of CYP1A-related ethoxyresorufin-O-deethylase (EROD). Furthermore, both CYP1A expression and EROD activity were further inhibited with increasing berberine concentrations. In addition, the CYP3A expressions at both the mRNA and the protein levels were downregulated by higher berberine concentrations. The catalytic activity of CYP3A-related erythromycin N-demethylase (ERND) was also inhibited by berberine at a dose of no less than 25mg/kg. Moreover, at the berberine concentration exceeding 25mg/kg, the inhibition of CYP3A expression and ERND activity increased with increasing berberine concentrations. In vitro experiments were also performed. When berberine was pre-incubated with the crucian carp liver microsomes, it competitively inhibited the corresponding EROD activity with the IC50 of 11.7μM. However, the ERND activity was slightly inhibited by berberine with the IC50 of 206.4μM. These results suggest that, in crucian carp, berberine may be a potent inhibitor to CYP1A, whereas the CYP3A inhibition needs a higher concentration of berberine.

Effects of cytochrome P450 1A substrate (difloxacin) on enzyme gene expression and pharmacokinetics in crucian carp ( hybridized Prussian carp)

Cytochrome P450s (CYPs) play a prominent role in drug metabolism and biotransformation which are distributed in liver of aquatic animals. However, limited information is available about CYP genes involved in drug metabolism in fish. In the present study, we explore CYP1A characterization for DIF metabolism. Firstly, we cloned and characterized the full-length cDNA sequence of a CYP1A gene from crucian carp ( hybridized Prussian carp), the predicted protein sequence for CYP1A comprise 496 amino acids. The heme-binding region of the CYP1A, encompassing the amino acid sequence GLGKRRCIG, which is identical to the same region of other homologues. Secondly, we studied the difloxacin (DIF) kinetics and the effects of DIF on their corresponding CYP1A mRNA levels in liver of crucian carp. CYP1A1 mRNA expression was analyzed by real-time PCR, and DIF concentration was determined by reversed-phase high-performance liquid chromatography (RP-HPLC). Results showed that the concentration of DIF in liver reached its peak (67.70 mg kg −1) at 0.5 h, while the CYP1A1 gene expression was at the lowest point. CYP1A mRNA was down-regulated by 6.5 mg ml −1 DIF in the liver of crucian carp. Thus, our work confirmed that DIF is both the substrate and inhibitor of CYP1A. The information provided a model for the potential utility of gene expression analysis and drug metabolization in fish.

Time-Dependent Oxidative Stress Responses of Crucian Carp (Carassius auratus) to Intraperitoneal Injection of Extracted Microcystins

This study was conducted to investigate time-dependent changes in oxidative enzymes in liver of crucian carp after intraperitoneally injection with extracted microcystins 600 and 150 microg kg(-1) body weight. The results showed that activities of antioxidant enzymes, including superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase generally exhibited a rapid increase in early phase (1-3 h post injection), but gradually decreased afterwards (12-48 h) compared with the control, with an evident time-dependent effect. These zigzag changes over time contributed a better understanding on oxidative stress caused by microcystins in fish.

Acute effects of microcystins on the transcription of antioxidant enzyme genes in crucian carp Carassius auratus

Recent evidences suggested that oxidative stress may play a significant role in the pathogenesis of MCs toxicity. In the present study, the acute effects of microcystins on the transcription of antioxidant enzyme genes were investigated in liver of crucian carp i.p.-injected with 50 mug MC-LReq per kg body weight (BW). We reported the cDNA sequences for four kinds of antioxidant enzyme (GSH-PX, CAT, Cu/Zn SOD, and GR) genes, and evaluated the oxidant stress induced by MCs through analyzing the transcription abundance of antioxidant enzyme genes using real-time PCR method. The time-dependent change of relative transcription abundance and expression of the antioxidant enzyme genes were determined at 1, 3, 12, 24, and 48 h. The transcription abundance varied among antioxidant enzymes, with GSH-PX and GR down-regulation, and CAT and SOD significantly upregulation. Based on these data, we tentatively concluded that the oxidant stress was induced by MCs, and caused the different response of the antioxidant enzyme genes.

Tissue Distribution and Depuration of the Extracted Hepatotoxic Cyanotoxin Microcystins in Crucian Carp (Carassius carassius) Intraperitoneally Injected at a Sublethal Dose

An acute toxicity experiment was conducted by intraperitoneal injection with a sublethal dose of extracted microcystins (MCs), 50 μg MC-LR (where L = leucine and R = arginine) equivalent/kg body weight (BW), to examine tissue distribution and depuration of MCs in crucian carp (Carassius carassius). Liver to body weight ratio increased at 3, 12, 24, and 48 h postinjection compared with that at 0 h (p < 0.05). MC concentrations in various tissues and aquaria water were analyzed at 1, 3, 12, 24, 48, and 168 h postinjection using liquid chromatography coupled with mass spectrometry (LC-MS). The highest concentration of MCs (MC-RR + MC-LR) was found in blood, 2–270 ng/g dry weight (DW), followed by heart (3–100 ng/g DW) and kidney (13–88 ng/g DW). MC levels were relatively low in liver, gonad, intestine, spleen, and brain. MC contents in gills, gallbladder, and muscle were below the limit of detection. Significant negative correlation was present between MC-RR concentration in blood and that in kidney, confirming that blood was important in the transportation of MC-RR to kidney for excretion. Rapid accumulation and slow degradation of MCs were observed in gonad, liver, intestine, spleen, and brain. Only 0.07% of injected MCs were detected in liver. The recovery of MCs in liver of crucian carp seemed to be dose dependent.