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Organochlorine residues in fish oil dietary supplements: comparison with industrial grade oils.

The market for fish oils as dietary supplements is of global importance. Although it is widely recognised that lipophilic organic chemicals, particularly organochlorines, can accumulate in fish oils, dietary supplements are not routinely considered when estimating average daily intakes for these contaminants. This paper reports levels of organochlorine residues in 44 fish oils, collected from 15 countries between 1994 and 1995, including 38 purchased over the counter as dietary supplements. Despite controls on the use of persistent organochlorine substances, appreciable quantities are found in oils sold as dietary supplements. Levels are discussed in relation to the significance of fish oil dietary supplements as contributors to daily intake of PCBs and pesticide residues.

Dietary Supplements↗

Contamination levels of PCDDs, PCDFs and CoPCbs in fish oils used as raw materials for margarine and shortening and retailed fish.

Toxic equivalent (TEQ) levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/DFs) and coplanar polychlorinated biphenyls (CoPCBs) were analyzed in both crude (n = 5) and refined-hardening fish oils (n = 2), which are used in part as the raw materials of margarine and shortening, and in some retailed fish (n = 6), which may be similar to those used as sources of fish oil. PCDDs/DFs-TEQ levels ranged from 0.1 to 13.5 pg TEQ/g fat weight in crude fish oils, from 0.5 to 1.6 pg TEQ/g fat weight in refined-hardening fish oils, and ranged from 1.4 to 31.2 pg TEQ/g fat weight in retailed fish. The ranges of PCDDs/DFs-TEQ in crude fish oils, in refined-hardening fish oils and in retailed fish overlapped each other. In addition, CoPCBs-TEQ levels ranged from 4.9 to 19.6 pg TEQ/g fat weight in crude fish oils, from 0.2 to 1.4 pg TEQ/g fat weight in refined-hardening fish oils, and from 2.7 to 165.8 pg TEQ/g fat weight in retailed fish. TEQ levels of CoPCBs in refined-hardening fish oils were much lower than those in crude fish oils and in retailed fish. This was due to the lower concentrations of less-chlorinated congeners in refined-hardening fish oils, as compared to those in crude fish oils and retailed fish. It appears that less-chlorinated PCBs congeners in refined-hardening fish oils are partly removed during the refining process.

Benzofurans↗

Up-regulation of liver uncoupling protein-2 mRNA by either fish oil feeding or fibrate administration in mice.

Fish oil feeding showed less obesity in rodents, relative to other dietary oils. N-3 fatty acids rich in fish oil and fibrate compounds are peroxisome proliferator-activated receptor alpha (PPARalpha) ligands that stimulate beta-oxidation of fatty acids in liver and are used for treatment of hypertriglycemic patients. Since UCP-2, a member of an uncoupling protein family, has been shown to express in hepatocytes, the effects of these agents on the expression of UCP2 mRNA were investigated. C57BL/6J mice were divided into three groups; the first group was given a high-carbohydrate diet, and the other two groups were given a high-fat diet (60% of total energy) as safflower oil or fish oil for 5 months. Safflower oil diet fed mice developed obesity, but those fed fish oil diet did not. Therefore, the effects of fish oil feeding on the expression of UCP1, UCP2 and UCP3 in liver, skeletal muscle (gastrocnemius), white adipose tissue (WAT) and brown adipose tissue (BAT) were assessed by Northern blotting. Compared with safflower oil feeding, fish oil feeding up-regulated liver UCP2, BAT UCP2 and skeletal muscle UCP3 mRNA, while down-regulated WAT UCP2 and BAT UCP3 mRNA. Among these alterations, 5-fold up-regulation of liver UCP2 mRNA, relative to carbohydrate feeding, was noteworthy. Fenofibrate administration (about 500 mg/kg BW/d) for 2 wks also induced liver UCP2 expression by 9-fold. These data indicated that fish oil feeding and fibrate administration each up-regulated UCP2 mRNA expression in liver possibly via PPARalpha and hence each has the potential of increasing energy expenditure for prevention of obesity.

Adipose Tissue↗

A comparison of dietary fish oil and corn oil in experimental colorectal carcinogenesis.

Rats fed either fish oil (n = 16) or corn oil (n = 16) in calorically and nutritionally balanced diets were injected with 1,2-dimethylhydrazine, which is a colorectal specific carcinogen; differential colorectal tumor induction was then measured. In addition, plasma peroxide concentrations were measured in rats in each dietary group as well as in a group receiving a low-fat diet, either with or without prior carcinogen treatment (n = 3 for each of the 6 groups). Tumor incidence did not differ between groups fed corn oil and fish oil. Tumor yield in the left colon was significantly lower in rats fed fish oil (p = 0.0499). Total colorectal tumors induced were also fewer in the rats fed fish oil (p = 0.065). Plasma peroxide concentrations were difficult to interpret because of the wide variation within groups. The data on tumor yield in the left colon support the hypothesis that a diet rich in n-3 fatty acids, which are found in fish oil, may be less supportive of colorectal tumor development than a diet rich in n-6 fatty acids, which is found in corn oil. However, the mechanism by which fish oil decreases tumor induction is still unknown.

1,2-Dimethylhydrazine↗

alpha-Tocopherol influences in vivo indices of lipid peroxidation in postmenopausal women given fish oil.

Although diets containing fish have been shown to be therapeutically valuable, the vitamin E requirement when large quantities of (n-3) fatty acids are consumed is not known. Additionally, as estrogens may function as an antioxidant, the requirement may be modified in postmenopausal women using hormone replacement therapy (HRT). Consequently, the purpose of this study was to measure the impact of graduated doses of RRR-alpha-tocopheryl acetate (TA) on in vivo indices of lipid peroxidation in postmenopausal women with and without hormone replacement therapy when given a supplement of fish oil. Forty-eight postmenopausal women, half receiving (+HRT) and half not receiving (-HRT) hormone replacement therapy, participated in a four-period, double-blind crossover trial. Each period lasted 5 wk followed by a 4-wk washout interval. During each period, the subjects consumed a 15-g supplement of fish oil and either 0, 100, 200, or 400 mg TA/d in a balanced, single square dosing order. Plasma levels of (n-3) fatty acids were significantly higher after fish oil supplementation; alpha-tocopherol concentration of plasma was significantly higher at each level of supplementation compared with the level without supplementation. Urinary excretion of thiobarbituric acid reactive substances (TBARS) and malondialdehyde, measured as the thiobarbituric-malondialdehyde adduct (TRA-MDA adduct), and the plasma concentration of the adduct were significantly greater after the fish oil supplement. Although urinary TBARS decreased linearly as the dose of TA increases (P < or = 0.05), urinary and plasma concentrations of TBA-MDA adduct did not. This study suggests that the evaluation of highly unsaturated fatty acids as oxidative stressors requires several measures of assessment.

Aged↗

Fish oil enhances pentachlorobenzene metabolism and reduces its accumulation in rats.

To investigate the influence of dietary fats on the metabolism and excretion of pentachlorobenzene (PECB), rats were fed semipurified diets containing soybean oil, lard or fish oil (10 g/100 g diet) for 2 wk. Rats then received a single dose of PECB by intragastric gavage. Blood concentration of pentachlorophenol (PCP), a major metabolite of PECB, was higher after 5 h through d 3 and that of PECB was lower after d 5 in the rats fed fish oil than in rats fed the other fats. After d 5, the highest ratio of PCP/PECB in the liver was seen in the fish oil-fed rats. Furthermore, fish oil intake resulted in a markedly lower concentration of PECB in fat tissues, where a large amount of PECB is typically distributed due to its lipophilic property. These findings indicate that fish oil feeding enhances PECB metabolism, thereby decreasing PECB residues in the body. Fish oil-fed rats had a smaller epididymal fat tissue mass compared with lard- or soybean oil-fed rats. The small mass of the fat tissues apparently limited the accumulation of PECB in those tissues and thus accelerated PECB metabolism in the liver.

Adipose Tissue↗

The improved survival of experimental animals fed with fish oil is suppressed by a leukotriene inhibitor.

Fish oil is believed to alter the immune response and improve survival after infections in experimental animals. This effect may be due to altered production of the leukotrienes (LT). We, therefore, performed a study in order to evaluate whether the effect of fish oil on the immune response of experimental animals is mediated through altered production of the LT. Female NMRI mice in four groups were fed with fish oil, fish oil with 5-lipoxygenase (5-LO) inhibitor (Zileuton, Abbott Laboratories, Chicago, IL, USA), corn oil or corn oil with 5-LO inhibitor. After 6 weeks, the mice were infected with Klebsiella pneumoniae and the survival was monitored. The experiment was performed twice. Analysis was performed mainly on data pooled from both experiments. The survival of the groups fed with fish oil was increased, compared to that of all the other groups and when compared to the groups fed with fish oil with 5-LO inhibitor (log-rank test) the difference was significant (P = 0.007). It has been postulated that the effect of fish oil on the immune system is mediated through altered production of LT. In our study, blocking of the production of the LT eliminated the beneficial effects of fish oil. Our results are in concord with the hypothesis that the effect of fish oil is, at least partly, mediated through altered production of LT.

Animals↗

Composition and properties of milk and butter from cows fed fish oil.

A control diet and a fish oil diet were fed to 12 multiparous Holstein cows to determine how the incorporation of Menhaden fish oil in the diet would influence the fatty acid composition, especially the conjugated linoleic acid and transvaccenic acid, contents of milk and butter. The control diet consisted of a 50:50 ratio of forage to concentrate, and the fish oil diet consisted of the control diet with 2% (on a dry matter basis) added fish oil. Milk from cows fed the control diet contained higher average concentrations of milk fat (3.37%) compared with milk from cows fed the fish oil diet (2.29%). Milk from cows fed fish oil contained higher concentrations of conjugated linoleic acid, transvaccenic acid, and total unsaturated fatty acids (0.68 and 2.51; 1.42 and 6.28; and 30.47 and 41.71 g/100 g of fat, respectively). Butter made from the fish oil diet milk also had higher concentrations of conjugated linoleic acid, transvaccenic acid, and unsaturated fatty acids. Penetrometer readings indicated fish oil diet butters were softer at 4 and 20 degrees C than the control diet butters. Acid degree values were similar in the fish oil butters compared with the control butters. No significant difference was found in the flavor characteristics of milk and butter from cows fed the control and fish oil diets. Production of milk and butter with increased amounts of conjugated linoleic acid, transvaccenic acid, and other beneficial fatty acids may have a desirable impact on the health of consumers and lead to increased sales.

Animals↗

Brain synaptosomal, liver, plasma, and red blood cell lipids in piglets fed exclusively on a vegetable-oil-containing formula with and without fish-oil supplements.

Clinical studies showed that a decrease in red blood cell 22:6n-3 caused by feeding infants formula (F) can be prevented by supplementation with fish oil (F + O). It is not known whether fish-oil supplementation is able to support normal accretion of fatty acids with greater than or equal to 20 carbons (LCPs) in the brain. Therefore piglets were fed exclusively F + O, F, or sow milk (SM) for 15 d and their liver and brain synaptosomal fatty acids were determined. Feeding F + O corrected the low n-3 LCP in the liver phospholipid (PL) and synaptosomal phosphatidylethanolamine (PE) of piglets fed F compared with SM. An apparent compensatory increase in n-6 LCPs in liver PL and synaptosomal PE of piglets fed F compared with SM was suppressed by feeding F + O. F + O also reduced the ratio of plasma PL 20:4n-6 to 20:5n-3, important for eicosanoid metabolism. Supplementation of F with n-3 LCPs as fish oil, without n-6 LCPs, at levels giving normal brain n-3 LCP, may alter n-6 LCP accretion.

Animals↗

Comparison of the effects of linseed oil and different doses of fish oil on mononuclear cell function in healthy human subjects.

Studies on animal and human subjects have shown that greatly increasing the amount of linseed (also known as flaxseed) oil (rich in the n-3 polyunsaturated fatty acid (PUFA) alpha-linolenic acid (ALNA)) or fish oil (FO; rich in the long-chain n-3 PUFA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)) in the diet can decrease a number of markers of immune function. The immunological effects of more modest doses of n-3 PUFA in human subjects are unclear, dose-response relationships between n-3 PUFA supply and immune function have not been established and whether ALNA has the same effects as its long-chain derivatives is not known. Therefore, the objective of the present study was to determine the effect of enriching the diet with different doses of FO or with a modest dose of ALNA on a range of functional responses of human monocytes and lymphocytes. In a randomised, placebo-controlled, double-blind, parallel study, forty healthy males aged 18-39 years were randomised to receive placebo or 3.5 g ALNA/d or 0.44, 0.94 or 1.9 g (EPA+DHA)/d in capsules for 12 weeks. The EPA:DHA ratio in the FO used was 1.0:2.5. ALNA supplementation increased the proportion of EPA but not DHA in plasma phospholipids. FO supplementation decreased the proportions of linoleic acid and arachidonic acid and increased the proportions of EPA and DHA in plasma phospholipids. The interventions did not alter circulating mononuclear cell subsets or the production of tumour necrosis factor-alpha, interleukin (IL) 1beta, IL-2, IL-4, IL-10 or interferon-gamma by stimulated mononuclear cells. There was little effect of the interventions on lymphocyte proliferation. The two higher doses of FO resulted in a significant decrease in IL-6 production by stimulated mononuclear cells. It is concluded that, with the exception of IL-6 production, a modest increase in intake of either ALNA or EPA+DHA does not influence the functional activity of mononuclear cells. The threshold of EPA+DHA intake that results in decreased IL-6 production is between 0.44 and 0.94 g/d.

Adolescent↗

Effects of dietary gamma-linolenic acid-rich borage oil combined with marine fish oils on tissue phospholipid fatty acid composition and production of prostaglandins E and F of the 1-, 2- and 3-series in a marine fish deficient in delta5 fatty acyl desaturase.

The effects of gamma-linolenic acid-rich borage oil (BO), in combination with different marine oils, namely an eicosapentaenoic acid (EPA) rich oil (MO) or a DHA-rich oil (TO), on tissue fatty acid composition and prostaglandin production were investigated in turbot, a species which lacks appreciable delta5 fatty acyl desaturase activity. The juvenile turbot grew well on the experimental diets and there were no significant differences in final weights between dietary treatments. Irrespective of the marine oil component, both the BO-containing diets increased tissue phospholipid levels of 18:2n-6 and 18:3n-6, and their respective elongation products, 20:2n-6 and 20:3n-6, compared to fish fed a control diet containing a standard Northern hemisphere fish oil. Both the BO-containing diets increased the production of 1-series prostaglandins (PG), this being observed across all tissues investigated with PGF and especially PGE. The BO/MO diet also reduced 20:4n-6 in tissue phospholipids without affecting 20:5n-3, whereas the BO/TO combination decreased 20:5n-3 but increased 20:4n-6. The production of 2-series and 3-series PGs was also altered by the dietary treatments but the changes were less dependent upon the tissue levels of their respective precursor fatty acids, 20:4n-6 and 20:5n-3. The BO-containing diets had very significant effects on gross fatty acid compositions of the phospholipids including increased proportions of saturated fatty acids and n-6 polyunsaturated fatty acids (PUFA) and decreased proportions of monounsaturated fatty acids and n-3 PUFA. Overall, this study shows that eicosanoid production in turbot tissues can be influenced by dietary fatty acids, not only by changes in the absolute and relative levels of specific eicosanoid precursor PUFA in tissue phospholipids, but also by general effects on membrane composition, structure and function induced by gross fatty acid compositional changes.

8,11,14-Eicosatrienoic Acid↗

Erythrocyte eicosapentaenoic acid versus docosahexaenoic acid as a marker for fish and fish oil consumption.

The fatty acid composition of erythrocyte membranes was investigated in 21 healthy men after 6 wk of varying intakes of eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3). In one experiment, 12 subjects were fed three diets in a 3 x 3 crossover design: an essentially fish-free control diet, a fish diet (0.15 g EPA/d, 0.41 g DHA/d) and the same fish-based diet supplemented with 5 g/d fish oil (Fish + Oil: 0.99 g EPA/d, 0.99 g DHA/d). A 6 wk wash-out period was allowed between each diet. In another experiment, 11 subjects were supplemented with 5 g/d fish oil alone for 6 wk (0.84 g EPA/d, 0.48 g DHA/d). After fish or fish oil feeding, the percent proportion of EPA and DHA in the erythrocyte membranes rose at the expense of linoleic and arachidonic acids. After 6 wk on the fish-based diets, EPA incorporation approached saturation, with the incremental increases being proportional to the amounts supplied by the diets. In contrast, parallel increases were observed for erythrocyte DHA even though the Fish + Oil diet was supplying twice as much DHA as the fish alone diet. These observations imply different metabolic rates for EPA and DHA and their importance is discussed in terms of the value of erythrocyte EPA versus DHA as markers for fish and fish oil consumption.

Adolescent↗

Fish and fish oil intake: effect on haematological variables related to cardiovascular disease.

To investigate the effects of practical amounts of n-3 highly-unsaturated fatty acids (HUFA) on a number of parameters involved in haemostasis, 12 healthy men were fed three diets in a 3 x 3 cross-over design. The diets, fed as the evening meal for 6-week periods, were: Control diet (essentially fish-free), Fish diet (200 g/d of lean Australian fish flesh) and the same fish-based diet but supplemented with 5 g/d fish oil (Fish + Oil). The diets supplied about 0, 0.6, and 2.0 g n-3 HUFA/d, respectively. Relative to the Control diet, the number of circulating leukocytes was significantly reduced after both the Fish and Fish + Oil diets (by 13% and 15%, respectively). This reduction occurred in the number of neutrophils, although this did not reach significance on the Fish diet alone. Platelet count fell on the Fish + Oil diet (by 6%) but not on the Fish diet alone. There was an apparent enhancement of fibrinolytic potential after both fish-containing diets, which tended to be accentuated with the fish oil supplementation. The mechanisms underlying some if not all of these observations may be eicosanoid-mediated as indicated by a diminution in the platelet arachidonic acid (20:4n-6) to eicosapentaenoic acid (20:5n-3) ratios after the Fish + Oil (13.7 +/- 1.8) and Fish (34.4 +/- 8.9) diets relative to the Control diet (66.1 +/- 15.6). These data suggest that a practical amount of lean fish can improve certain haematological parameters implicated in the etiology of cardiovascular disease.

Animals↗

Potential of thraustochytrids to partially replace fish oil in Atlantic salmon feeds.

The replacement of fish oil with a dried product made from thraustochytrid culture, a marine microorganism, in canola-oil-based diets for Atlantic salmon was investigated. Salmon (37 g) were fed for 51 days on diets containing only canola oil, canola oil and fish oil, or canola oil and the thraustochytrid. There were no significant differences in final weight (106.1 +/- 1.1 g), weight gain (69.6 +/- 1.1 g), feed consumption (16.5 +/- 0.2 mg dry matter g(-1) d(-1)), feed efficiency ratio (1.15 +/- 0.03 g (g-1)), or productive protein value (51.2% +/- 1.7%) between the diets. Nor were there any significant differences in whole-body chemical composition, organ somatic indices, or measures of immune function. However, following transfer to seawater and 2 challenges with Vibrio anguillarum, cumulative mortality was significantly lower in fish fed some fish oil than in those fed the 2 diets containing no fish oil. In conclusion, the thraustochytrid had no detrimental effects on the performance of salmon but, at the current inclusion of 10%, failed to confer the same effect as fish oil under challenging conditions.

Animal Nutritional Physiological Phenomena↗

Plasma triacylglycerol and coagulation factor concentrations predict the anticoagulant effect of dietary fish oil in overweight subjects.

Fish oil, containing (n-3) PUFA, is associated with a moderate reduction in cardiovascular disease through a multifactorial mechanism involving a decrease in plasma lipids and anticoagulant activity. Two intervention studies on subjects at risk were performed to determine the relation of these 2 fish-oil effects. In study 1, 54 overweight subjects consumed 3.1 g (n-3) PUFA daily. In study 2, which involved 42 overweight patients with type 2 diabetes, 20 subjects consumed (n-3) PUFA, whereas 22 others ingested a preparation rich in (n-6) PUFA. Tissue factor-induced thrombin generation (thrombin potential) was determined as an integrated measure of plasma coagulant activity. In both studies, multivariate analysis indicated a strong clustering of fasting concentrations of triacylglycerols, prothrombin, factor V, factor VII, and factor X with one another at baseline. This cluster of factors determined partly the interindividual variation in thrombin generation, of which prothrombin and triacylglycerol concentrations were the main determinants. In both healthy subjects and diabetes patients, high triacylglycerol concentrations (>1.69 mmol/L) at baseline were closely linked to a strong fish oil-induced lowering of triacylglycerol and coagulation factor V, VII, and X concentrations, and thrombin generation. We conclude that high fasting triacylglycerol concentrations predict high procoagulant activity and a lowering of thrombin potential with dietary fish oil.

Adult↗

A comparative study on the effect of algal and fish oil on viability and cell proliferation of Caco-2 cells.

Polyunsaturated fatty acid (PUFA) rich micro-algal oil was tested in vitro and compared with fish oil for antiproliferative properties on cancer cells in vitro. Oils derived from Crypthecodinium cohnii, Schizochytrium sp. and Nitzschia laevis, three commercial algal oil capsules, and menhaden fish oil were used in cell viability and proliferation tests with human colon adenocarcinoma Caco-2 cells. With these tests no difference was found between algal oil and fish oil. The nonhydrolysed algal oils and fish oil showed a much lower toxic effect on cell viability, and cell proliferation in Caco-2 cells than the hydrolysed oils and the free fatty acids (FFAs). Eicosapentaenoic acid (EPA; C20:5n-3) and docosahexaenoic acid (DHA; C22:6n-3) were used as samples for comparison with the tested hydrolysed and nonhydrolysed oils. The hydrolysed samples showed comparative toxicity as the free fatty acids and no difference between algal and fish oil. Oxidative stress was shown to play a role in the antiproliferative properties of EPA and DHA, as alpha-tocopherol could partially reverse the EPA/DHA-induced effects. The results of the present study support a similar mode of action of algal oil and fish oil on cancer cells in vitro, in spite of their different PUFA content.

Antioxidants↗

Modulation of lipid profile by fish oil and garlic combination.

Fish consumption has been shown to influence epidemiology of heart disease, and garlic has been shown to influence triglyceride levels. This study was undertaken to evaluate the effect of fish oil and garlic combinations as a dietary supplement on the lipid subfractions. Forty consecutive subjects with lipid profile abnormalities were enrolled in a single-blind, placebo-controlled crossover study. Each subject received placebo for 1 month and fish oil (1800 mg of eicosapentanoic acid [EPA] + 1200 mg of docosahexanoic acid) with garlic powder (1200 mg) capsules daily for 1 month. Lipid fractionation was performed prior to study initiation, after the placebo period, and after the intervention period. Subjects all had cholesterol levels > 200. Subjects were instructed to maintain their usual diets. Supplementation for 1 month resulted in an 11% decrease in cholesterol, a 34% decrease in triglyceride, and a 10% decrease in low-density lipoprotein (LDL) levels, as well as a 19% decrease in cholesterol/high-density lipoprotein (HDL) risk. Although not significant, there was a trend toward increase in HDL. There was no significant placebo effect. These results suggest that in addition to the known anticoagulant and antioxidant properties of both fish oil and garlic, the combination causes favorable shifts in the lipid subfractions within 1 month. Triglycerides are affected to the largest extent. The cholesterol lowering and improvement in lipid/HDL risk ratios suggests that these combinations may have antiatherosclerotic properties and may protect against the development of coronary artery disease.

Adult↗

Effect of fish oil diet on immune response and proteinuria in mice.

In this study, we examined the immune response and proteinuria caused by dietary polyunsaturated fatty acids in normal NZW/N and autoimmune NZB/NZW mice. Mice were maintained more than one year on five dietary groups: normal (5% corn oil), calorie-restricted, high fat (20% corn oil), high fat (20% fish oil), and Purina laboratory rodent chow. Normal mice fed with the fish oil diet had a more reduced anti-sheep red blood cells (SRBC) plaque-forming cell (PFC) response and less interleukin-2 (IL-2) enhancement of PFC than did the group with the restricted diet and the young control group. The corn oil (5 and 20%) diet animals also showed reduced PFC response and IL-2 utilization. NZB/NZW mice fed with the fish oil diet showed similar reduced PFC response but had a significantly lower response to IL-2 than did those on the corn oil diets and the restricted diet. The IL-2 production by macrophages from NZW/N mice was reduced in both the fish oil and corn oil diet groups. However, mice fed with the fish oil diet had less proteinuria and good survival rates, similar to the group with the restricted diet. These results suggest that the beneficial effect of the fish oil diet in these animals may be attributed in part to the immunosuppression mechanism.

Animals↗