Management of Hypercholesterolemia Through Dietary ß-glucans-Insights From a Zebrafish Model.

Consumption of lipid-rich foods can increase the blood cholesterol content. ß-glucans have hypocholesterolemic effect. However, subtle changes in their molecular branching can influence bioactivity. Therefore, a comparative investigation of the cholesterol-lowering potential of two ß-glucans with different branching patterns and a cholesterol-lowering drug, namely simvastatin was undertaken employing the zebrafish (Danio rerio) model of diet-induced hypercholesterolemia. Fish were allocated to 5 dietary treatments; a control group, a high cholesterol group, two ß-glucan groups, and a simvastatin group. We investigated plasma total cholesterol, LDL and HDL cholesterol levels, histological changes in the tissues, and explored intestinal transcriptomic changes induced by the experimental diets. Dietary cholesterol likely caused the suppression of endogenous cholesterol biosynthesis, induced dysfunction of endoplasmic reticulum and mitochondria, and altered the histomorphology of the intestine. The two ß-glucans and simvastatin significantly abated the rise in plasma cholesterol levels and restored the expression of specific genes to alleviate the endoplasmic reticulum-related effects induced by the dietary cholesterol. Furthermore, the distinct patterns of transcriptomic changes in the intestine elicited by the oat and microalga ß-glucans impacted processes such as fatty acid metabolism, protein catabolic processes, and nuclear division. Oat and microalgal ß-glucans also altered the pattern of lipid deposition in the liver. Our study provides insights into the effectiveness of different ß-glucans to alleviate dysfunctions in lipid metabolism caused by dietary cholesterol.

Lens metabolomic profiling as a tool to understand cataractogenesis in Atlantic salmon and rainbow trout reared at optimum and high temperature.

Periods of high or fluctuating seawater temperatures result in several physiological challenges for farmed salmonids, including an increased prevalence and severity of cataracts. The aim of the present study was to compare cataractogenesis in Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss) reared at two temperatures, and investigate whether temperature influences lens metabolism and cataract development. Atlantic salmon (101±2 g) and rainbow trout (125±3 g) were reared in seawater at either 13°C (optimum for growth) or 19°C during the 35 days experiment (n = 4 tanks for each treatment). At the end of the experiment, the prevalence of cataracts was nearly 100% for Atlantic salmon compared to ~50% for rainbow trout, irrespective of temperature. The severity of the cataracts, as evaluated by slit-lamp inspection of the lens, was almost three fold higher in Atlantic salmon compared to rainbow trout. The global metabolic profile revealed differences in lens composition and metabolism between the two species, which may explain the observed differences in cataract susceptibility between the species. The largest differences were seen in the metabolism of amino acids, especially the histidine metabolism, and this was confirmed by a separate quantitative analysis. The global metabolic profile showed temperature dependent differences in the lens carbohydrate metabolism, osmoregulation and redox homeostasis. The results from the present study give new insight in cataractogenesis in Atlantic salmon and rainbow trout reared at high temperature, in addition to identifying metabolic markers for cataract development.

Atlantic salmon (Salmo salar) require increased dietary levels of B-vitamins when fed diets with high inclusion of plant based ingredients.

Aiming to re-evaluate current recommendations for nutrient supplementations when Atlantic salmon are fed diets based on plant ingredients, two regression experiments, with parr and post-smolt, were conducted. A control diet was included to evaluate if ingredients supplied sufficient nutrients without any added nutrient package (NP). The nutrient package consisted of vitamins B, C, E, minerals, cholesterol, methionine, taurine and histidine. This paper focus on B-vitamins. In parr, growth, health and welfare parameters responded on NP additions, but this was not observed in the seawater stage. During three months of feeding, parr tripled their weight. Parr given diets added the NP above NRC (2011) showed improved protein retention, and reduced liver and viscera indices. Post-smolt fed the same diets during five months showed a doubling of weight, but did not respond to the variation in NP to the same extent as parr. Significant regressions were obtained in body compartments for several of the B-vitamins in the premix. Whole body biotin concentration was unaffected by micronutrient premix level, and mRNA expression of the enzymes dependent of biotin showed only weak increases with increased biotin. Muscle thiamine plateaued at a diet level similar to NRC (2011) recommendation in freshwater, and showed stable values independent on premix addition in seawater. The mRNA expression of the enzyme G6PDH (glucose-6-phosphate dehydrogenase) is sensitive to thiamine availability; results did not indicate any need to add thiamine above levels recommended for fish in general. Niacin showed a steady increase in whole body concentrations as feed niacin increased. Muscle riboflavin peaked at a diet level of 12.4 mg kg-1. Sufficient riboflavin is important to avoid e.g., development of cataract. Cataract was not registered to be any problem, neither in fresh- nor in seawater. Cobalamin (B 12) in muscle and liver was saturated at 0.17 mg kg-1 diet. Muscle pyridoxine showed a dose-dependent level in muscle, and peaked around 10 mg kg -1 diet. White muscle ASAT (asparagine amino transferase) activity steadily increased, with indications of stable values when dietary pyridoxine was around 10-16 mg kg -1 diet. Pantothenic acid increased in gill tissue up to a level of 5.5 mg kg -1 soft gill tissue; at a dietary level of 22 mg kg-1. Improved performance, and coverage of metabolic need for niacin was at a dietary level of 66 mg kg -1, riboflavin 10-12 mg kg-1, pyridoxine 10 mg kg-1 and panthotenic acid 22 mg kg-1. Based on these results, recommended B-vitamin supplementation in plant based diets for Atlantic salmon should be adjusted.

Aryl hydrocarbon receptor protein and Cyp1A1 gene induction by LPS and phenanthrene in Atlantic cod (Gadus morhua) head kidney cells.

The objective of this study was to evaluate interactions between environmental toxicants and cod immune cells during inflammation. Phenanthrene is abundant in plant oils (rapeseed, palm, and soya oil) as compared to fish oils, and consequently constitute an undesirable element in plant replacement diets in aquaculture. Phenanthrene was added to head kidney cell cultures, alone or together with LPS (lipopolysaccharide) or poly I: C (polyinosinic acid: polycytidylic acid), and the responses were evaluated in terms of protein and gene expression. The results showed that LPS, poly I: C or phenanthrene, added to the cultures separately, induced aryl hydrocarbon receptor (AhR) protein expression. Phenanthrene treatment in combination with LPS induced AhR protein expression and Cyp1A1 gene transcription, which not was observed combining poly I: C and phenanthrene. Phenanthrene exposure up regulated the transcription of common stress and detoxification enzymes like catalase, caspase 3 and glutathione S-transferase alfa 3 subunit B (GSTAB3), while LPS exposure alone or combined with phenanthrene down regulated GSTAB3 and catalase in cod leukocytes. It seems clear that immune regulation and phenanthrene induced signaling pathways interact; transcriptional down regulation of detoxification and antioxidant enzymes by LPS could indicate that combating bacterial infections is the number one priority in these cells, and that AhR and Cyp1A1 is somehow involved in this signaling cascade. LPS seems to affect the mitogen activated protein kinases (MAPKs) pathways (P-p38 and ERK1/2) thus modulating the AhR protein and Cyp1A1 gene transcription, while phenanthrene possibly activates AhR by ligand binding.

Pathogen recognition and mechanisms in Atlantic cod (Gadus morhua) head kidney cells: bacteria (LPS) and virus (poly I:C) signals through different pathways and affect distinct genes.

Understanding pathogen recognition and mechanisms in Atlantic cod are of significant importance for both basic research on wild populations and health management in aquaculture. A microarray approach was utilized to search for effects of viral (polyinosinic acid:polycytidylic acid), bacterial (lipopolysaccharide) and polyclonal activator (phytohaemoagglutinin) stress in Atlantic cod head kidney cells. LPS cell activation increased mRNA expression of interleukin 8; interleukin-1ß; cyclooxygenase 2; leukocyte derived chemotaxin 2; carboxyl-esterase 2 and environmental biomarker cytochrome P450 1A. Mitogen activated protein kinase p38 and cathepsin F were down regulated by LPS. The antiviral responses induced by double stranded RNA clearly increased transcription of Toll like receptor 3 and interferon stimulating gene 15. The phytohaemoagglutinin response seemed to be more non-specific. Special for the phytohaemoagglutinin induction was the increase in major histocompatibility complex class I. CC chemokine type 2 mRNA expression was increased by phytohaemoagglutinin, lipopolysaccharide and polyinosinic acid:polycytidylic acid, while mitogen activated protein kinase p38 and leukocyte derived chemotaxin 2 were down regulated by phytohaemoagglutinin. Oxidative stress related genes like catalase and glutaredoxin and the anti-apoptotic gene Bcl-2 showed no transcriptional changes compared to control in any of the treatments. Eicosanoids like prostaglandin 2, leukotriene B4 and B5 were constitutively produced by cod head kidney cells in vitro. The most remarkable feature of eicosanoid secretion is the higher production of leukotrienes against prostaglandins, indicating that the lipooxygenase pathway is preferred over the cyclooxygenase pathway. Although there were no significant differences in eicosanoid secretion between the groups, polyinosinic acid:polycytidylic acid showed a clear tendency to increase the levels of leukotriene B4 and B5. This study reveals distinct signatures of bacteria and virus transcriptional responses in cod head kidney cells. In addition, the novel finding that cytochrome P450 1A was upregulated during the antibacterial response indicates a connection between immunity and aryl hydrocarbon receptor activation in Atlantic cod.

Induction and activity of oxidative stress-related proteins during waterborne Cd/Zn-exposure in brown trout (Salmo trutta).

We studied how transcript levels of metallothionein (MT), Cu/Zn-superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GR) as well as functional protein levels of MT, SOD and CAT in brown trout tissues changed during a 15-days waterborne exposure to Cd and Zn. Trout from a river with low levels of metals (the Stribekken River) was transferred to a river with high levels of Cd and Zn (the Naustebekken River) and exposed up to 15 days. The aim of this transfer experiment was to investigate how exposure to Cd and Zn induced transcription and activities of central antioxidant enzymes and proteins in an environmental setting. Significant uptake of both Cd and Zn was observed in gills during the 15 days exposure, and Cd levels was found to correlate significantly with transcript levels of MT-A, SOD, GPx and GR. Gill concentrations of Zn did not correlate significantly with the transcript levels of the stress genes studied, but Zn might have triggered transcription of proteins which dealt with subsequent accumulation of Cd. SOD and CAT activities increased in gills after transfer, but MT protein levels decreased. In liver, SOD activity and MT protein levels increased, while in kidney only MT protein concentrations were elevated after transfer. There was a general lack of consistency between mRNA transcription and enzyme activities, indicating that these proteins and enzymes are not solely under transcriptional control.