Body surface area-based omega-3 fatty acids supplementation strongly correlates to blood concentrations in children.

Omega-3 fatty acids have been suggested as a complement in cancer treatment, but doses are not established. We performed a dose-finding study in 33 children in remission from cancer. Participants were allocated to a body surface area (BSA) adjusted dose (mg/m2) of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (40:60), ranging 233-3448 mg/m2 daily for 90 days. Fatty acid concentration in plasma phospholipids and red blood cells were determined by GC. Supplementation was well tolerated and correlated strongly with blood ω3-fatty acid concentrations and EPA showed the highest increase. Using the ω3-index disregards docosapentaenoic acid (DPA), which increased 30-43% in our study motivating an EDD-index (∑EPA,DPA,DHA). The ratio between arachidonic acid and EPA or DHA showed negative exponential trends. Dose per BSA enabled an individualized omega-3 supplementation decreasing the variation referred to interindividual differences. Based on our results, we suggest a dose of 1500 mg/m2 BSA for further studies.

Feeding steam-pelleted rapeseed affects expression of genes involved in hepatic lipid metabolism and fatty acid composition of chicken meat.

This study investigated the dietary effect of steam-pelleted rapeseed (RS) diets with different inclusion levels on the fatty acid composition of chicken meat and the expression of lipid metabolism-related genes in the liver. Experimental diets included 6 different wheat-soybean meal based diets either in nonpelleted or steam-pelleted form supplemented with 80, 160, and 240 g RS/kg feed and one nonpelleted wheat-soybean meal based diet without RS supplementation as the control. These diets were fed to newly hatched broiler chickens (Ross 308) for 34 days. Compared to the control diet, steam-pelleted diets containing 160 or 240 g/kg RS significantly increased the content of omega-3 long chain polyunsaturated fatty acids (n-3 LC-PUFA) in the breast and drumstick, while their meat yields were not affected. Moreover, the mRNA levels of fatty acid desaturase 1 (FADS1) and acyl-coenzyme A oxidase 1 (ACOX1) in their livers increased. Therefore, steam-pelleted diets with 160 or 240 g/kg RS can be used to increase the n-3 LC-PUFA content in chicken meat without compromising meat yield.

Intake of carp meat from two aquaculture production systems aimed at secondary prevention of ischemic heart disease - a follow-up study.

Our previous study showed that a diet enriched with 400 g of carp per week improved plasma lipids in subjects after aortocoronary bypass (CABG). The aim of the present study is to determine whether the different carp farming systems have an impact on the effects of carp meat in secondary cardiovascular prevention. We examined 3 groups of patients after CABG over a 4-week period of spa treatment (108 persons, 73 males, 35 females, age over 60 years). We found no differences in baseline values of blood pressure or plasma lipids. The patients were given a standard spa diet (controls; N=36) or a diet enriched of 400 g of carp meat per week, enriched omega 3 (N=37) or cereal carp (N=35). Plasma lipid parameters were examined at start and after 4 weeks in a routine laboratory setting. Group consuming omega-3 carp showed the largest decline in total cholesterol, LDL cholesterol, triglycerides and an increase in HDL cholesterol (all p<0.01). We found that carp meat from the two production systems showed significantly different effects on plasma lipids. Further trials should be performed to clarify the exact causes of the differences.

Influence of oilseed supplement ranging in n-6/n-3 ratio on fatty acid composition and Δ5-, Δ6-desaturase protein expression in steer muscles.

This study investigated effects of roasted or extruded oilseed supplementation ranging in n-6/n-3 ratios from 0.3 to 5.0 on the fatty acid composition and expression of delta-5 desaturase (Δ5d) and Δ6-desaturase (Δ6d) protein in commercial steer cheek (m. masseter) and diaphragm (pars costalis diaphragmatis) muscles. In general, the n-6/n-3 ratio of the diet had a subsequent effect on the muscle n-6/n-3 ratio (P < 0.05), with muscle 18:2n-6 and 18:3n-3 content relating to proportion of dietary soya bean and linseed (P < 0.01). Compared with canola, pure linseed and soya bean diets reduced 14:1c-9 and 16:1c-9 (P < 0.05) but increased 18:1t-11 and c-9,t-11 conjugated linoleic acid (CLA) content (P < 0.01). Oilseed processing had a minor influence but extruded oilseeds increase 18:1t-11 and c-9,t-11 CLA compared with roasted (P < 0.05). Polar lipid 18:3n-3 and n-3 long-chain polyunsaturated fatty acid (LC, ⩾20 carbons PUFA) derivative content increased in relation to dietary linseed supplementation in the diaphragm (P < 0.01), whereas only 18:3n-3 was increased in the cheek (P < 0.01). Protein expression did not differ between diets; however, in each muscle the Δ5d protein expression had a stronger association with the desaturase products rather than the precursors. The relationship between Δ5d protein expression and the muscle LC n-6/n-3 ratio was negative in both muscles (P < 0.05). The relationship between Δ6d protein expression and the LC n-6/n-3 ratio was positive in the cheek (P < 0.001) and negative in the diaphragm (P < 0.05). In conclusion, diet n-6/n-3 ratio affected muscle 18:2n-6 and 18:3n-3 deposition, whereas the Δ5d and Δ6d protein expression had some influence on the polar lipid LC-PUFA profile. Results reaffirm that processed oilseeds can be used to increase the proportion of fatty acids potentially beneficial for human health, by influencing the formation of LC-PUFA and reducing the n-6/n-3 ratio.

Sesamin modulates gene expression without corresponding effects on fatty acids in Atlantic salmon (Salmo salar L.).

This study examined the effects of sesamin inclusion in vegetable oil-based diets fed to Atlantic salmon (Salmo salar L.). The diets used differed in n-6/n-3 fatty acid (FA) ratio (0.5 and 1) and sesamin content (high 5.8 g/kg, low 1.16 g/kg and no sesamin). The oils used in the feeds were a mixture of rapeseed, linseed and palm oil. Fish were fed for 4 months. Fatty acids and expression of hepatic genes involved in transcription, lipid uptake, desaturation, elongation and ß-oxidation were measured. No major effects on the percentage of DHA in white muscle, liver triacylglycerol and phospholipid fraction were detected. Genes involved in ß-oxidation, elongation and desaturation were affected by sesamin addition. Limited effects were seen on any of the transcription factors tested and no effect was seen on the expression of peroxisome proliferator-activated receptors (PPAR). Expression of both SREBP-1 and SREBP-2 increased with sesamin addition. It was concluded that supplementation of fish feed with a high level of sesamin had a negative effect on the growth rate and live weight and did not alter the proportions of DHA in tissues even though gene expression was affected. Thus, more studies are needed to formulate a diet that would increase the percentage of DHA in fish without negative effects on fish growth.

Association of DGAT1 genotype, fatty acid composition, and concentration of copper in milk with spontaneous oxidized flavor.

In 136 cows with altogether 969 milk samples, we investigated the effect of the acyl-coenzyme A:diacylglycerol acyltransferase 1 (DGAT1) K232A polymorphism on milk fatty acid (FA) composition and how, in combination with copper concentration in milk, this influences the occurrence of spontaneous oxidized flavor. All milk samples were analyzed for concentrations of copper and individual FA and subjected to sensory analysis by trained judges. We found an effect of DGAT1 genotype on FA composition where mainly the long-chain FA were affected. The 232A allele was associated with larger proportions of the C18:2 cis-9,trans-11 conjugated linoleic acid and lower proportions of C16:0 FA. Milk concentrations of unsaturated FA and copper showed strong and unfavorable associations with spontaneous oxidized flavor (SOF) development. The interaction between FA and copper indicates that SOF will not develop as easily in milk with high copper content unless the substrate is available (i.e., in addition to the previously shown effect of copper in milk, unsaturated FA are required for the process of oxidation to progress). We observed a marked effect of the DGAT1 genotype on SOF development where the A allele was associated with a higher risk of SOF. Moreover, our results suggest that the effects of the FA C18:3 n-3 and of the polyunsaturated index on SOF development are beyond the effect of the DGAT1 genotype. Breed had an effect on FA composition but not on SOF development. Our results imply that copper, FA composition, and DGAT1 genotype are risk factors for SOF and considerations to these factors might be necessary in future breeding decisions.

Verapamil does not modify catalytic activity of CYP450 in rainbow trout after long-term exposure.

Little is known about the effects of the cardiovascular drug verapamil (VRP) on metabolic processes in fish. Most calcium channel blockers including VRP are metabolized by cytochrome P450 (CYP450) enzymes. In this study we investigated the in vivo effect of VRP on some CYP450-mediated reactions in juvenile rainbow trout (Oncorhynchus mykiss). Fish were exposed to sublethal concentrations of VRP (0.5, 27 and 270 µg l(-1)) for 0, 21, and 42 day. The following CYP450-mediated reactions were studied in hepatic microsomes: O-dealkylation of ethoxyresorufin, methoxyresorufin, and pentoxyresorufin, hydroxylation of coumarin, tolbutamide, and p-nitrophenol, and O-debenzylation of 7-benzyloxy-4-trifluoromethylcoumarin. The amounts of products of these reactions did not differ among fish exposed to different levels of VRP and control fish. This suggests that the levels of VPR used did not alter catalytic activity of the selected CYP450 enzymes. In conclusion, none of the investigated CYP450-mediated reactions has potential as a biomarker to monitor VRP contamination of the aquatic environment.

Influence of dietary sesamin, a bioactive compound on fatty acids and expression of some lipid regulating genes in Baltic Atlantic salmon (Salmo salar L.) juveniles.

The effects of inclusion of sesamin / episesamin in Baltic Atlantic salmon (Salmo salar L.) diets based on vegetable oils were studied. The study was designed as a dose response study with two control diets, one diet based on fish oil (FO) and one diet based on a mixture of linseed and sunflower oil (6:4 by vol.) (MO). As experimental diets three different levels of inclusion of sesamin / episesamin (hereafter named sesamin) to the MO based diet and one diet based on sesame oil and linseed oil (SesO) (1:1 by vol.) were used. The dietary oils were mirrored in the fatty acid profile of the white muscle. Sesamin significantly decreased the levels of 18:3n-3 in the white muscle phospholipid (PL) fraction of all groups fed sesamin, no significant differences were found in the triacylglycerol fraction (TAG). Slightly increased levels of docosahexaenoic acid (22:6n-3, DHA) in PL and TAG were found in some of the sesamin fed groups. Sesamin significantly affected the expression of peroxisome proliferator-activated receptor alpha, scavenger receptor type B and hormone sensitive lipase, in agreement with previous studies on rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar L.) hepatocytes published by our group. No significant effects on toxicological response measured as ethoxyresorufin O-deethylase activity was found. The total cytochrome P450 enzymes were significantly higher in MO 0.29 and SesO group. The amount of alpha- and gamma-tocopherols in liver and the amount of gamma-tocopherol in white muscle were significantly lower in fish fed the FO diet compared to the MO diet, but no difference after inclusion of sesamin was found in this study. Increased inclusion of sesamin increased the levels of sesamin and episesamin in the liver, but did not affect the amounts in white muscle.

Fatty acid transfer from sow to piglet differs for different polyunsaturated fatty acids (PUFA).

Polyunsaturated fatty acids (PUFA) are essential for the development of the nervous system in animals. It is known that pigs are good models for human in many aspects. The aim of the study was to investigate how fat content and FA composition in sows' diet influence FA composition in brain of newborn and in liver and brain of one-day-old piglets, respectively. High fat (6 %) feeds were designed with regard to saturated or polyunsaturated fat content and n-6/n-3 ratio by adding either oats rich in linoleic acid (LA) or linseed oil rich in alpha-linolenic acid (ALA). The ratio n-6/n-3 PUFA was 11 in all three diets (the low fat (3 %), high fat saturated and high fat oats diet), while the ratio in the linseed oil diet was 2. Increased proportion of ALA in the diet increased ALA and eicosapentaenoic acid (EPA) in piglets' neutral and polar liver lipids and the long chain PUFA, EPA, docosapentaenoic and docosahexaenoic acid in piglet brain. The results suggest that transport of n-3 PUFA from sow to piglet was higher via milk than via bloodstream in the uterus and that increased content of ALA in sows' feed led to an increased accumulation of n-3 FA in piglets' liver and brain.

Influence of vitamins A, D3 and E status on post-mortem meat quality in steers under winter housing or pasture finishing systems.

We investigated the influence of Swedish recommended vitamins A, D3 and E supplementation levels on muscle tenderness and fatty acid (FA) composition under indoor or outdoor finishing programmes. Swedish Red breed steer calves were divided into vitamin supplemented (n = 12) and non-supplemented (n = 15) groups while on pasture prior to the finishing period. This trial began at the beginning of the winter housing period during which the steers were fed a 55 : 45 dry matter barley : grass silage diet indoors. The indoor finished group was comprised of vitamin supplemented (n = 6) and non-supplemented (n = 8) steers slaughtered after about 155 days on feed. Vitamin supplemented steers were provided with 100 g mineral supplement providing 400 000 IU vitamin A, 100 000 IU D3 and 3000 IU E daily as recommended for Swedish production practices. In spring, outdoor finished vitamin supplemented (n = 6) and non-supplemented (n = 7) steers grazed semi-natural grassland for an additional 120 days before slaughter. During pasture, vitamin supplemented steers had free-choice access to a mineral supplement containing vitamins A, D3 and E. The mineral supplement for the non-supplemented steers did not contain vitamins A, D3 and E and was provided at the same amount as the vitamin supplemented steers. Shear force values were similar between vitamin supplemented and non-supplemented steers after ageing 2, 7 and 14 days within indoor and outdoor finishing programmes. The shear force values had decreased by 14 days of ageing within all programmes. The µ- and m-calpain activity did not differ between vitamin supplemented and non-supplemented steers for either the indoor or outdoor finishing programmes. The calpastatin activity was higher for the indoor, vitamin supplemented steers. Indoor finished vitamin supplemented steers had a greater proportion of C18:1c-9 and total monounsaturated fatty acids, whereas the non-supplemented steers had a greater proportion of total saturated fatty acids. We concluded that the meat quality from steers not receiving vitamin supplementation was similar to that of steers receiving vitamins A, D3 and E supplementation at Swedish recommended levels under indoor and outdoor finishing programmes.

Importance of knowledge on lipid composition of foods to support development towards consumption of higher levels of n-3 fatty acids via freshwater fish.

The need of better labelling of fats in processed animal origin products is urgent. The lack of information makes it possible to exclude n-3 fatty acids in preparations of foods. The higher fat content, the higher n-6/n-3 ratio seems to be a rule. It is desirable to broaden the labelling into which oils have been used when foods are processed. The dietary balance of n-6 and n-3 fatty acids is important for homeostasis and normal development in humans. The ratio between n-6/n-3 fatty acids suggested to be evolutionary developed is between 1 and 4. The main conclusion is that the fat sources used during processing and preparation of convenient foods have the largest impact on the food FA content and composition. A proposal is therefore that this should be declared on the product label especially the n-3 FA content. It is also of large importance to increase consumption of freshwater fish fed suitable feeds containing n-3 fatty acids in central Europe to enable a generally lower n-6/n-3 ratio in the human diet. Therefore optimizing feeds to freshwater fish in culture is urgent and important.

Differences between lipid content and composition of different parts of fillets from crossbred farmed carp (Cyprinus carpio).

Common carp (Cyprinus carpio) is, globally, one of the most important farmed fish species. We have analyzed carp from a natural pond-production system in South Bohemia. Ongrowing fish had access to an abundant plankton population which was supplemented with cereals. Fatty acid composition (FA) was investigated in three parts of fillets from four crosses of carp. The FA composition of the leanest part, the dorsal white muscle was similar to that of marine-farmed species; it contained a large proportion of n-3 highly unsaturated FA, the n-3/n-6 ratio was 1.1, and the proportion of phospholipids was high. The abdominal wall is rich in monounsaturated FA, the n-3/n-6 ratio is lower, 0.5, and it is more affected by the cereal feed. We concluded that the lipid composition of all these carp tissues can be improved in terms of healthy FA profile.

Effects of crude rapeseed oil on lipid composition in Arctic charr Salvelinus alpinus.

This study investigated the effects of crude rapeseed oil (RO) on lipid content and composition in muscle and liver of Arctic charr Salvelinus alpinus. Triplicate groups were fed diets containing fish oil (FO):RO ratio of 100:0, 75:25, 50:50 and 25:75 until two-fold mass increase. Total lipid content increased significantly in the liver with higher proportion of RO in the diet. Profound effects were seen in the fatty acid composition in the analysed tissues with a reduction in 20:5n-3 and 22:6n-3 and an increase in 18:2n-6 with higher RO content in the diets. A drop in cholesterol content was seen at 25% inclusion of RO in both tissues. Wild-caught fish contained a considerably higher amount of 20:4n-6 in both storage and membrane lipids of white muscle compared with the experimental fish.

Sesamin increases alpha-linolenic acid conversion to docosahexaenoic acid in atlantic salmon (Salmo salar L.) hepatocytes: role of altered gene expression.

In vitro cultivated Atlantic salmon (Salmo salar L.), hepatocytes were incubated without or with a mixture of sesamin and episesamin in order to test for possible effects on lipid metabolism. Sesamin/episesamin exposure (0.05 mM, final concentration) led to increased elongation and desaturation of (14)C 18:3n-3 to docosahexaenoic acid ((14)C 22:6n-3, DHA, P < 0.01) and down regulated gene expression of Delta6 and Delta5 desaturases compared to control treatment. Sesamin/episesamin further increased the hepatocytes capacity for fatty acid beta-oxidation of (14)C 18:3n-3 (P < 0.01) to the (14)C acid soluble products, acetate, malate and oxaloacetate, in agreement with an increased gene expression of carnitine palmitoyltransferase I. Also the gene expression of cluster of differentiation 36 was upregulated and the expression of scavenger receptor type B, peroxisome proliferator-activated receptors alpha and gamma were downregulated. The amount of triacylglycerols secreted by the cells tended to be lower in the sesamin/episesamin incubated hepatocytes than the control cells. This study shows that sesamin has favourable effects on lipid metabolism leading to increased level of DHA, which may be of interest for aquaculture use.

Sesamin supplementation increases white muscle docosahexaenoic acid (DHA) levels in rainbow trout (Oncorhynchus mykiss) fed high alpha-linolenic acid (ALA) containing vegetable oil: metabolic actions.

The effects of including an equi-mixture of sesamin and episesamin in fish diets based on vegetable oils of different fatty acid composition were examined. Sesamin/episesamin (hereafter named sesamin) was included at 0.58 g/100 g diet. The oil used in the feed was either a mixture of linseed and sunflower oils (6:4, by vol) or 100% linseed oil. Addition of sesamin increased the percentages of docosahexaenoic acid (DHA) in white muscle phospholipid and triacylglycerol fraction by up to 37% but the fatty acids in red muscle and liver were not affected. The expression of the peroxisome proliferator-activated receptor PPARalpha was significantly down regulated in the liver of the fish fed sesamin and mixed oil diet (P < 0.05). Sesamin and episesamin were detected in liver and muscle tissues of the fish that had been fed sesamin. Fish fed sesamin had elevated levels of total cytochrome P450 (CYP) enzymes and EROD activity in the liver, indicating an induction of CYP1A in this tissue. Our conclusion was that supplementation of fish feed with sesamin increased the proportions of DHA in the white muscle.

Influence of diet on fatty acids and tocopherols in M. longissimus dorsi from reindeer.

Our aim was to compare the effects of two pelleted diets containing differing FA composition with natural lichen pasture on reindeer's meat FA composition. In addition we wanted to increase the knowledge about reindeer FA metabolism and the effect of animal sex and age on FA composition in reindeer muscle. The trial included five reindeer groups: three grazing, consisting of adult males, adult females, and calves; and two groups of calves fed conventional pellets (CPD) and pellets containing crushed linseed (LPD), respectively, for two months before slaughter. Differences between male and female animals were mainly found in the neutral lipid fraction and related to fatness. Calves differed significantly from adult males and females in FA and lipid class composition. CPD led to a higher ratio of n-6 to n-3 FA compared with grazing. The ratio n-6/n-3 in the polar lipid (PL) fraction of the animals fed LPD was slightly, but not significantly, higher than that in the grazing reindeer. LPD-fed animals had lower proportions of long-chain polyunsaturated FA (LCPUFA), namely 20:4n-6, 22:5n-3, and 22:6n-3, in the PL fraction compared with the grazing animals due to the content of these FA in the natural feed. The animals seemed unable to elongate dietary FA in significant amounts. We conclude that by adding crushed linseed to the pellets it was possible to keep the favorable FA composition of meat from grazing reindeer with regard to the n-6/n-3 ratio but not in LCPUFA.

Fatty acids, antioxidants and oxidation stability of processed reindeer meat.

The effect of hot smoking and drying on reindeer meat (M. semimembranosus) was investigated with regard to vitamin content, fatty acids (FA), lipid oxidation and lipolysis. In smoked and dried meat, free fatty acids were found to increase (P<0.001), whereas polar lipids, cholesterol and triacylglycerols decreased. Only slight changes in FA composition of the smoked meat were found but the composition of the dried meat differed significantly from both smoked and fresh meat. Fresh and smoked meat were found to have low values (0.11 µg/g dry matter (DM); 0.21 µg/g DM) of thiobarbituric acid reacting substances but dried meat had high values (8.33 µg/g DM; P<0.001). Retinol was found only in the fresh meat and the tocopherol content decreased significantly (P<0.001) as a result of the processing. We conclude that the smoking process slightly changed FA composition, lipid class composition and vitamin content, whereas drying resulted in major changes in all the analysed parameters.

Fatty acid composition and tocopherol concentrations in muscle of entire male, castrated male and female pigs, reared in an indoor or outdoor housing system.

The objective of this study was to investigate the effect of outdoor rearing on the fatty acid (FA) composition and contents of tocopherols of M. longissimus dorsi polar (PL) and neutral lipids (NL) in entire males, castrated males and female pigs. Fat content did not differ between genders or rearing conditions. In PL of the indoor pig muscle the level of n-6 highly unsaturated fatty acids (HUFA) was higher in females and entire males compared with castrated males. The outdoor environment resulted in a slightly higher level of n-3 polyunsaturated FA (PUFA) in muscle PL and NL of entire males compared with females and castrated males. We also found an increased level of 18:2 n-6 in the muscle PL of outdoor reared females compared with the indoor females. The higher levels of PUFA in the entire male muscle, in both PL and NL, were not accompanied by a higher level of vitamin E.

Muscle lipids, vitamins E and A, and lipid oxidation as affected by diet and RN genotype in female and castrated male Hampshire crossbreed pigs.

The objectives of this study were to investigate the polar and neutral lipid fatty acid composition, content of retinol, α-tocopherol, γ-tocopherol and level of oxidation of the pig muscle (M. longissimus dorsi). Female and castrated male Hampshire crossbreeds were produced in two systems. One group was raised indoors with a more polyunsaturated diet and the other raised outdoors with a more saturated diet. The level of polyunsaturated fatty acids in muscle was higher in the indoor females compared with the outdoor females, indoor castrated males and outdoor castrated males. The increased level of polyunsaturated fatty acids in the muscle, which was accompanied by a relatively low content of α-tocopherol, increased the susceptibility to lipid oxidation in the form of MDA (malondialdehyde) in the indoor female pigs. Finally, the level of polyunsaturated fatty acids in the polar lipids was affected by the RN genotype, and this difference was dependent on sex. In conclusion, diet has a major effect on the fatty acid composition and oxidation stability in pork muscle, but additional factors such as sex and RN genotype might also contribute.