Freshwater Fish Crucian Carp Research Articles

The dynamic effects of different inorganic arsenic species in crucian carp (Carassius auratus) liver during chronic dietborne exposure: Bioaccumulation, biotransformation and oxidative stress

Inorganic arsenic (iAs) is highly toxic to aquatic species, but the chronic effect of iAs on fish following dietborne exposure is still unclear. In this study, freshwater fish crucian carp (Carassius auratus) was exposed to iAs [arsenite (AsIII) and arsenate (AsV)] for 40 days through dietary exposure. The bioaccumulation and biotransformation of arsenic in the main metabolic organ, liver, were measured. The oxidative stress responses to iAs exposure in liver were analyzed to be linked to arsenic biotransformation, especially methylation. In both AsIII and AsV groups, the total As contents gradually increased during the exposure and then fleetly decreased at the end of exposure (40 d). Arsenobetaine was found to be the predominated As species (34–66%) and the fraction remained on an increasing trend, while the inorganic As percentages decreased 84–91% during the 40-day exposure, suggesting that the capability of As biotransformation increased to acclimate iAs during chronic dietborne exposure. Both the activities of the enzymatic antioxidants (superoxide dismutase and catalase) and the level of the nonenzymatic antioxidant (glutathione) increased initially and then decreased, thus lowering the malondialdehyde levels and displaying a typical antioxidant defense mechanism. The opposite correlations were observed between arsenic secondary methylation index and the malondialdehyde level in different iAs treatment. This indicated that the As dimethylation played an significant role toward oxidative damage; the toxic action of As dimethylation was dependent upon the parent iAs species at the initial stage of exposure. Therefore, the effectiveness of the detoxification relied on both the biomethylation rate of As and the anti-oxidation ability based on nonenzymatic antioxidant and enzymatic antioxidant.

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 in the crucian carp (Carassius auratus) exposed to environmentally relevant concentration of 17α-Ethinylestradiol based on metabolomics

17α-ethynylestradiol (EE2), a ubiquitous synthetic endocrine disrupting chemical, was the principal component of contraceptive drugs and one of common hormone medications. The detrimental impact of EE2 on the reproduction of organisms was widely recognized. However, the underlying mechanisms of physiological and metabolome effects of EE2 on freshwater fish are still unclear. This study investigated the toxic effects and related mechanisms of EE2 on freshwater fish crucian carp (Carassius auratus) based on metabolomics. Crucian carp were exposed to EE2 at environmentally relevant concentration for 9 days, 18 days, and 27 days, and the biological responses were explored through analysis of the physiological endpoints, steroid hormones, and metabolome. The physiological endpoints of crucian carp had no distinct change after EE2 exposure. However, metabolomics analysis probed significant deviation based on chemometrics, indicating that the metabolomics approach was more sensitive to the effects of EE2 at environmentally relevant concentration to freshwater fish than the traditional endpoints. The alterations of 24 metabolites in gonad and 16 metabolites in kidney were induced by treatment with EE2, respectively, which suggesting the perturbations in amino acid metabolism, lipid metabolism, energy metabolism, and oxidative stress. Moreover, EE2 exposure could induce the disruption of lipid metabolism and then broke the homeostasis of endogenous steroid hormones. Metabolomics provided a new strategy for the studies on contaminant exposure at a low dose in the short term and gave important information for the toxicology and mechanism of EE2.

Oxygen- and temperature-dependent expression of survival protein kinases in crucian carp (Carassius carassius) heart and brain.

Living without oxygen is limited to very few vertebrates, one species being the fresh water fish crucian carp (Carassius carassius), which can survive months of anoxia at low temperatures. Mammalian heart and brain are particularly intolerant to oxygen deprivation, yet these organs can be conditioned to display increased resistance, possibly due to activation of several protein kinases. We hypothesized increased phosphorylation status of these kinases in hypoxic and anoxic crucian carp heart and brain. Moreover, we wanted to investigate whether the kinases showing the strongest phosphorylation during hypoxia/anoxia, ERK 1/2, p38-MAPK, JNK, PKCε, and PKCδ, also had increased expression and phosphorylation at cold temperatures, to better cope with the anoxic periods during winter. We found small differences in the phosphorylation status of ERK 1/2, p38-MAPK, JNK, PKCε, and PKCδ during 10 days of severe hypoxia in both heart and brain (0.3 mg O₂/l) and varying responses to reoxygenation. In contrast, 7 days of anoxia (<0.01 mg O₂/l) markedly increased phosphorylation of ERK 1/2, p38-MAPK, JNK in the heart, and p38-MAPK and PKCε in the brain. Similarly, varying acclimation temperature between 4, 10 and 20°C induced large changes in phosphorylation status. Total protein expression in heart and brain neither changed during different oxygen regimes nor with different acclimation temperatures, except for ERK 1/2, which slightly decreased in the heart at 4°C compared with 20°C. A phylogenetic analysis confirmed that these protein kinases are evolutionarily conserved across a wide range of vertebrate species. Our findings indicate important roles of several protein kinases during oxygen deprivation.

Investigation of the genotoxicity of endosulfan to freshwater Cyprinid fish Crucian carp (Carassius carassius L.) using the micronucleus and chromosomal aberration as biomarkers

This study was aimed to evaluate the in vivo genotoxic effects of endosulfan in freshwater fish crucian carp, Carassius carassius L. The LC50–96h value of endosulfan (with 95 % confidence limits), determined by probit analysis, was 0.070 (0.046–0.093) ppm L−1 and the estimated safe levels of endosulfan varied from 00.70 × 10−2 to 00.70 × 10−6 ppm L−1. Based on the LC50–96h value, three test concentrations (viz. sub-lethal I, II, and III) were determined to be 0.052, 0.035 and 0.017 ppm L−1, respectively, and the fish specimens were exposed to these concentrations for 35 days. The mean concentration of endosulfan in water samples (aquaria) was always within 5 % of the intended concentration when analyzed by dispersive liquid-liquid micro extraction (DLLME) followed by gas chromatography–mass spectrometry (GC-MS). Autopsy was done on days 1, 2, 3, 4, 7, 14, 21, 28 and 35 of endosulfan exposure for assessment of micronucleus (MN) formation and chromosomal aberrations (CA). The MN formation in the peripheral erythrocytes, authenticated by scanning electron microscopy (SEM), was found to be significantly higher (p < 0.05) in all the treated groups, including positive control cyclophosphamide (4 ppm L−1) compared to negative control. Similar significant effects (p < 0.05) were also observed on CA in the head kidney cells. In general, both MN and CA exhibited a concentration and time-dependent response. The current study revealed the potential genotoxicity of endosulfan in fish; and that the micronucleus and chromosomal assays are useful tools in determining genotoxicity of water pollutants and might be appropriate as a part of environmental monitoring program.