Dietary vitamin A supplementation ameliorates the effects of poly-aromatic hydrocarbons in Atlantic salmon (Salmo salar).

Several studies have reported on the interaction between vitamin A (VA) and aryl hydrocarbon receptor (AhR)-binding toxicants, including poly-aromatic hydrocarbons (PAHs). In aquaculture, the use of plant oils in novel aquafeeds can increase PAH levels while simultaneously lowering natural VA background levels, causing the need to supplement plant oil-based feeds with synthetic VA. To study dietary VA-PAH interactions, Atlantic salmon (initial weight 195±0.15g) were fed four identical plant-based diets that were supplemented with PAHs (100 and 10mgkg(-1) benzo[a]pyrene (BaP) and phenanthrene (Phe), respectively) or VA (retinyl acetate 8721IUkg(-1)) separately or combined for 2.5 months in a 2×2 factorial design, with triplicate net-pens per diet. Dietary PAH significantly reduced hepatic VA storage, and VA-enriched diets restored hepatic VA. There was a significant PAH-VA interaction effect on hepatic BaP, but not Phe, accumulation, with reduced hepatic BaP concentrations in fish fed VA+PAH compared to fish fed PAH alone. Concurrently, PAH and VA significantly interacted in their effects on CYP1A phase I biotransformation as observed from increased ethoxyresorufin-O-deethylase (EROD) activity, increased CYP1A protein concentration, and elevated transcription (cyp1a1 gene expression) in fish fed PAH+VA compared to PAH alone. Dietary VA supplementation alone had no significant effect on CYP1A phase I biotransformation. Metabolomic assessment showed that dietary VA caused a restoration of metabolic intermediates involved in energy metabolism that were affected by dietary PAH. Moreover, a PAH-induced growth inhibition was partially ameliorated by dietary VA supplementation. In conclusion, dietary VA interacted with PAH toxicity on the level of CYP1A-mediated detoxification, hepatic PAH accumulation, energy allocation, and growth.

In vitro Assessment of Hg Toxicity in Hepatocytes from Heat-Stressed Atlantic Salmon.

Global warming may alter the bioavailability of contaminants in aquatic environments. In this work, mercury (Hg2+) toxicity was studied in cells obtained from Atlantic salmon smolt kept at 15 °C (optimal growth temperature) for 3 months or at a stepwise increase to 20 °C (temperature-stress) during 3 months prior to cell harvest to evaluate whether acclimation temperature affects Hg toxicity. To examine possible altered dietary requirements in warmer seas, one group of fish following the stepwise temperature regimes was fed a diet spiked with antioxidants. Atlantic salmon hepatocytes were exposed in vitro to 0, 1.0, or 100 µM Hg2+ for 48 h. Cytotoxicity, determined as electrical impedance changes with the xCELLigence system, and transcriptional responses, determined with RT-qPCR, were assessed as measures of toxicity. The results showed that inorganic Hg at a concentration up to 100 µM is not cytotoxic to Atlantic salmon hepatocytes. Significance and directional responses of the 18 evaluated target genes suggest that both Hg and temperature stress affected the transcription of genes encoding proteins involved in the protection against ROS-generated oxidative stress. Both stressors also affected the transcription of genes linked to lipid metabolism. Spiking the diet with antioxidants resulted in higher concentrations of Se and vitamin C and reduced concentration of Hg in the liver in vivo, but no interactions were seen between the dietary supplementation of antioxidants and Hg toxicity in vitro. In conclusion, no evidence was found suggesting that inorganic Hg is more toxic in cells harvested from temperature-stressed fish.

Modifying effects of vitamin E on chlorpyrifos toxicity in atlantic salmon.

The aim of this study was to elucidate how vitamin E (alpha tocopherol) may ameliorate the toxicity of the pesticide chlorpyrifos in Atlantic salmon. Freshly isolated hepatocytes were exposed to vitamin E, chlorpyrifos or a combination of vitamin E and chlorpyrifos (all 100 µM). Transcriptomics (RNA-seq) and metabolomics were used to screen for effects of vitamin E and chlorpyrifos. By introducing vitamin E, the number of upregulated transcripts induced by chlorpyrifos exposure was reduced from 941 to 626, while the number of downregulated transcripts was reduced from 901 to 742 compared to the control. Adding only vitamin E had no effect on the transcriptome. Jak-STAT signaling was the most significantly affected pathway by chlorpyrifos treatment according to the transcriptomics data. The metabolomics data showed that accumulation of multiple long chain fatty acids and dipeptides and amino acids in chlorpyrifos treated cells was partially alleviated by vitamin E treatment. Significant interaction effects between chlorpyrifos and vitamin E were seen for 15 metabolites, including 12 dipeptides. The antioxidant had relatively modest effects on chlorpyrifos-induced oxidative stress. By combining the two data sets, the study suggests that vitamin E supplementation prevents uptake and accumulation of fatty acids, and counteracts inhibited carbohydrate metabolism. Overall, this study shows that vitamin E only to a moderate degree modifies chlorpyrifos toxicity in Atlantic salmon liver cells.

Endosulfan in vitro toxicity in Atlantic salmon hepatocytes obtained from fish fed either fish oil or vegetable oil.

The composition of the feed may alter the cellular composition of an organism and thus has the potential to influence a xenobiotic response. The main aim of this study was to see if the fatty acid composition of primary hepatocytes isolated from Atlantic salmon (Salmo salar L.) obtained from fish fed either a fish oil or a vegetable oil based diet, influenced the response to endosulfan exposure in vitro. The primary cultures were exposed to six different concentrations of endosulfan (0.001, 0.01, 0.1, 1, 10 and 100 microM) for 48 h. Cell morphology as well as a molecular toolbox of 16 genes encoding stress responsive and biotransformation proteins was examined. Endosulfan exposure caused moderate cytotoxicity and steatosis in a dose-dependent manner in the hepatocytes. In general, endosulfan hepatoxicity seems to be unaffected by the fatty acid composition of the hepatocytes. Exceptions were general stress (HSP70) and markers for estrogen exposure (ZP and VTG), which appeared to be slightly less responsive in hepatocytes isolated from the vegetable oil fed fish.