Dietary lipid quality and mitochondrial membrane composition in trout: responses of membrane enzymes and oxidative capacities

To examine whether membrane fatty acid (FA) composition has a greater impact upon specific components of oxidative phosphorylation or on overall properties of muscle mitochondria, rainbow trout (Oncorhynchus mykiss) were fed two diets differing only in FA composition. Diet 1 was enriched in 18:1n-9 and 18:2n-6 while Diet 2 was enriched in 22:6n-3. The FA composition of mitochondrial phospholipids was strongly affected by diet. 22:6n-3 levels were twice as high (49 %) in mitochondrial phospholipids of fish fed Diet 2 than in those fed Diet 1. 18:2n-6 content of the phospholipids also followed the diets, whereas 18:1n-9 changed little. All n-6 FA, most notably 22:5n-6, were significantly higher in fish fed Diet 1. Nonetheless, total saturated FA, total monounsaturated FA and total polyunsaturated FA in mitochondrial phospholipids varied little. Despite a marked impact of diet on specific FA levels in mitochondrial phospholipids, only non-phosphorylating (state 4) rates were higher in fish fed Diet 2. Phosphorylating rates (state 3), oxygen consumption due to flux through the electron transport chain complexes as well as the corresponding spectrophotometric activities did not differ with diet. Body mass affected state 4 rates and cytochrome c oxidase and F ₀ F ₁ ATPase activities while complex I showed a diet-specific effect of body mass. Only the minor FA that were affected by body mass were correlated with functional properties. The regulated incorporation of dietary FA into phospholipids seems to allow fish to maintain critical membrane functions even when the lipid quality of their diets varies considerably, as is likely in their natural environment.     

N-3 HUFAs affect fat deposition,susceptibility to oxidative stress,and apoptosis in Atlantic salmon visceral adipose tissue

We have investigated how n-3 highly unsaturated fatty acids (HUFAs) in the diet affect fatty acid (FA) utilization, fat storage and oxidative stress (OS) in Atlantic salmon (Salmo salar) white adipose tissue (WAT). Four groups of Atlantic salmon were fed for 21 weeks on one of the four diets supplemented with 23% (of dry matter) lipid. Docosahexaenoic acid (DHA; 22:6n-3) and eicosapentaenoic acid (EPA; 20:5n-3) levels increased from 10% of total FAs in the rapeseed oil (RO) diet, to 20% in the fish oil (FO) diet, and to 50% and 55% in the DHA-enriched and EPA-enriched diets, respectively. Increased dietary levels of n-3 HUFAs resulted in lower fat percentage in WAT. Furthermore, mitochondrial FA β-oxidation activity was higher in the FO group than it was in the RO group. The relative levels of DHA and EPA in phospholipids (PLs) from WAT and mitochondrial membranes increased with the increasing dietary levels of these HUFAs. In general, the mitochondrial membrane PLs were characterised by lower relative levels of n-3 HUFAs and higher relative levels of linoleic acid (LA; 18:2 n-6) than WAT membrane PLs. The predominance of LA relative to n-3 HUFAs in mitochondrial membrane PLs may help to protect these PLs from peroxidation. Cytochrome c oxidase measurements revealed higher incidence of disrupted mitochondrial membranes in the DHA and EPA dietary groups than in the FO and RO dietary groups. This disruption further affected the mitochondrial function, resulting in a marked reduction in FA β-oxidation capacities. The reduction in mitochondrial function and the increase in the activity of superoxide dismutase (SOD) in the DHA and EPA groups showed that high dietary dose of DHA and EPA resulted in oxidative stress (OS). The increased activity of caspase 3 in the high n-3 HUFA groups suggested the induction of apoptosis and increased incidence of cell death in WAT, which may be one of the factors explaining the lower fat percentage found in these groups.     

Changes in biochemical and hemocyte parameters of the Pacific oysters Crassostrea gigas fed T-Iso supplemented with lipid emulsions rich in eicosapentaenoic acid

The aim of this study was to assess the effect of dietary eicosapentaenoic acid (20:5n-3) on hemocyte parameters such as hemocyte concentration, phagocytosis, and non-stimulated reactive oxygen species (ROS) production in Pacific oysters Crassostrea gigas, as well on proximate biochemical and fatty acid compositions. One-year-old oysters (C. gigas) were fed T-Isochrysis aff. galbana (T-Iso), which is low in 20:5n-3, either alone or with supplements of a lipid emulsion rich in 20:5n-3 at 1%, 10% or 50% (dry weight of the algal ration) for up to 7 weeks. Changes in gill fatty acid composition demonstrated that the lipid emulsion was well ingested by oysters during the dietary conditioning. Biochemical analysis indicated that oysters fed supplements of 50% and, to a lesser extent, 10% lipid emulsions had a higher total lipid content compared with oysters fed other diets, suggesting a more advanced reproductive status for the oysters fed high doses of lipid emulsion. Moreover, some oysters in these two treatment groups spawned during the last three weeks of the seven-week feeding experiment. Lipid supplements had a significant influence on hemocyte concentration, phagocytic index and non-stimulated hemocyte ROS production. After 4 weeks, highest hemocyte concentrations were found in oysters fed on a supplement of 50% lipid emulsion compared with those fed on other diets but the hemocytes derived from these oysters had the lowest short-term phagocytic index. After 7 weeks of dietary conditioning, the ROS production in non-stimulated hemocytes of oysters fed 10% and 50% lipid emulsion declined. These results suggested that 20:5n-3, and perhaps its eicosanoid metabolites, affected oyster hemocyte functions; however, the reproductive status of oysters may also have interfered with the 20:5n-3 dietary effect.     

Dietary fatty acid composition changes mitochondrial phospholipids and oxidative capacities in rainbow trout red muscle

Dietary conditioning of juvenile trout changed the acyl chain composition of mitochondrial phospholipids and the oxidative capacities of muscle mitochondria. Trout were fed three diets differing only in fatty acid (FA) composition. The highly unsaturated 22:6 n-3 (DHA) accounted for 0.4, 14, and 30% of fatty acids in Diets 1, 2 and 3. After 10 weeks of growth, the dietary groups differed markedly in FA composition of mitochondrial phospholipids, with significant dietary effects for virtually all FA. Mean mitochondrial DHA levels were 19, 40 and 33% in trout fed Diets 1, 2 and 3. Mitochondrial oxidative capacities changed with diet, while mitochondrial concentrations of cytochromes and of the adenylate nucleotide translocase (nmol mg¹ protein) did not. Mitochondria from fish fed Diet 1 had higher non-phosphorylating (state 4) rates at 5°C than those fed other diets. When phosphorylating (state 3) rates differed between dietary groups, rates at 5 and 15°C were higher for fish fed the more unsaturated diets. Stepwise multiple regressions indicated that FA composition could explain much (42–70%) of the variability of state 4 rates, particularly at 5°C. At 15°C, FA composition explained 16–42% of the variability of states 3 and 4 rates. Similar conclusions were obtained for the complete data set (trout fed diets 1, 2 and 3) and for the data from trout achieving similar growth rates (e.g. those fed Diets 1 and 2). Neither general characteristics of membrane FA, such as % saturates, unsaturation index, n-3, n-6 or n-3/n-6 nor levels of abundant unsaturated FA such as DHA or 18:1(n-9 + n-7), were systematically correlated with mitochondrial capacities even though they differed considerably between trout fed the different diets. Relatively minor FA (20:5n-3, 20:0, 18:2n-6, 18:3n-3, 18:0 and 15:0) showed better correlations with mitochondrial oxidative capacities. This supports the concept that acyl chain composition modulates mitochondrial capacities via interactions between membrane proteins and specific FA of particular phospholipid classes in their microenvironment.     

Potential use of seaweeds in the laying hen ration to improve the quality of n-3 fatty acid enriched eggs

An association between dietary fish oil and decreased yolk weight and reduced sensory quality of eggs has been reported when eggs are enriched with n-3 FA from fish oil. Seaweeds are an important source of compounds that seem to increase egg weight when included in the laying hen diet. The objectives of this study were to determine the influence of the dietary seaweeds Macrocystis pyrifera, Sargassum sinicola and Enteromorpha sp. on the physical quality, lipid composition and consumer acceptability of n-3 FA enriched eggs. One-hundred and forty-four 35-week-old Leghorn hens were randomly distributed in four treatments that consisted of the inclusion of 2% of sardine oil (SO) and 10% of each marine alga (MA) in laying hens' diets; a control diet (C) was also prepared. The study lasted 8 weeks and egg physical quality, egg lipids and sensory attributes were evaluated. The results showed that incorporation of 10% M. pyrifera in the diets is an effective way of increasing the n-3 FA content, the albumen height and yolk color, but not the egg weight, when these are enriched with n-3 FA from fish oil. The egg flavor was also not affected.     

The use of stoned olive cake and rolled linseed in the diet of intensively reared lambs: effect on the intramuscular fatty-acid composition

The aim of the present study was to evaluate the effect of the inclusion of stoned olive cake and rolled linseed in a concentrate-based diet for lambs on the fatty-acid composition of polar and non-polar intramuscular lipids of the longissimus dorsi muscle. To achieve this objective, 32 Appenninica lambs were randomly distributed into four groups of eight lambs each and were fed conventional cereal-based concentrates (diet C); concentrates containing 20% on a dry matter (DM) basis of rolled linseed (diet L); concentrates containing 35% DM of stoned olive cake (diet OC); and concentrates containing both rolled linseed (10% DM) and stoned olive cake (17% DM; diet OCL). The concentrates were administered together with grass hay at a 20:80 forage:concentrate ratio. Growing performances and carcass traits were evaluated. The fatty-acid composition was analysed in the total intramuscular lipids, as well as in the polar and neutral lipids. The average feed intake and the growth performance of lambs were not affected by the dietary treatments, as a consequence of similar nutritional characteristics of the diets. The inclusion of rolled linseed in the L and OCL diets increased the content of C18:3 n-3 in intramuscular total lipids, which was threefold higher in meat from the L lambs and more than twofold higher in meat from the OCL lambs compared with the C and OC treatments. The n-6:n-3 ratio significantly decreased in the meat from lambs in the L and OCL groups, reaching values below 3. The L treatment resulted in the highest level of trans-18:1 fatty acids in the muscle. Regardless of the dietary treatment, the t10-18:1 was the major isomer, representing 55%, 45%, 49% and 45% of total trans-18:1 for C, L, OC and OCL treatments, respectively. Neutral lipids from the OC-fed lambs contained the highest amount of c9-18:1 (more than 36% of total fatty acids); however, the content of c9-18:1 did not differ between the OC and C lambs, suggesting an intensive biohydrogenation of dietary c9-18:1 in the case of OC treatment. The highest content of c9,t11-18:2 was detected in the intramuscular fat from the L-fed lambs, followed by the OCL treatment. A similar trend was observed in the neutral lipid fraction and, to a lower extent, in the polar lipids.     

Effects of olive and fish oil Ca soaps in ewe diets on milk fat and muscle and subcutaneous tissue fatty-acid profiles of suckling lambs

Enhancing healthy fatty acids (FAs) in ewe milk fat and suckling lamb tissues is an important objective in terms of improving the nutritional value of these foods for the consumer. The present study examined the effects of feeding-protected lipid supplements rich in unsaturated FAs on the lipid composition of ewe milk, and subsequently in the muscle and subcutaneous adipose tissues of lambs suckling such milk. Thirty-six pregnant Churra ewes with their new-born lambs were assigned to one of three experimental diets (forage/concentrate ratio 50 : 50), each supplemented with either 3% Ca soap FAs of palm (Control), olive (OLI) or fish (FO) oil. The lambs were nourished exclusively by suckling for the whole experimental period. When the lambs reached 11 kg BW, they were slaughtered and samples were taken from the Longissimus dorsi and subcutaneous fat depots. Although milk production was not affected by lipid supplementation, the FO diet decreased fat content (P<0.001), whereas the OLI milk FA profile resembled that of the Control diet. In contrast, although FO drastically diminished the contents of stearic and oleic acids (P<0.001), all the saturated even-numbered carbon FAs from 6:0 to 14:0 increased (P<0.05). FO also produced the highest levels of c9,t11-18:2 (2.21%) and n-3 FAs, 20:5 n-3 (0.58%), 22:5 n-3 (0.48%) and 22:6 n-3 (0.40%). The high levels of trans-11 18:1 (7.10%) obtained from the FO diet would suggest that Ca soaps only confer partial protection in the rumen. In contrast, the lack of significant differences in trans-10 18:1 levels (P>0.05) and other trans-FAs between Control and FO treatments would indicate that FO treatment does not alter rumen biohydrogenation pathways under the assayed conditions. Changes in dam milk FA composition induced differences in the FA profiles of meat and fat depots of lambs, preferentially incorporated polyunsaturated FAs into the muscle rather than storing them in the adipose tissue. In the intramuscular fat of the FO treatment, all the n-3 FAs reached their highest concentrations: 0.97 (18:3 n-3), 2.72 (20:5 n-3), 2.21 (22:5 n-3) and 1.53% (22:6 n-3). In addition, not only did FO intramuscular fat have the most cis-9, trans-11 18:2 (1.66%) and trans-11 18:1 (3.75%), but also the lowest n-6/n-3 ratio (1.80) and saturated FA content were not affected. Therefore, FO exhibited the best FA profile from a nutritional point of view.     

Evaluation of the effects of different diets on microbiome diversity and fatty acid composition of rumen liquor in dairy goat

Fat supplementation plays an important role in defining milk fatty acids (FA) composition of ruminant products. The use of sources rich in linoleic and α-linolenic acid favors the accumulation of conjugated linoleic acids isomers, increasing the healthy properties of milk. Ruminal microbiota plays a pivotal role in defining milk FA composition, and its profile is affected by diet composition. The aim of this study was to investigate the responses of rumen FA production and microbial structure to hemp or linseed supplementation in diets of dairy goats. Ruminal microbiota composition was determined by 16S amplicon sequencing, whereas FA composition was obtained by gas-chromatography technique. In all, 18 pluriparous Alpine goats fed the same pre-treatment diet for 40±7 days were, then, arranged to three dietary treatments consisting of control, linseed and hemp seeds supplemented diets. Independently from sampling time and diets, bacterial community of ruminal fluid was dominated by Bacteroidetes (about 61.2%) and Firmicutes (24.2%) with a high abundance of Prevotellaceae (41.0%) and Veillonellaceae (9.4%) and a low presence of Ruminococcaceae (5.0%) and Lachnospiraceae (4.3%). Linseed supplementation affected ruminal bacteria population, with a significant reduction of biodiversity; in particular, relative abundance of Prevotella was reduced (−12.0%), whereas that of Succinivibrio and Fibrobacter was increased (+50.0% and +75.0%, respectively). No statistically significant differences were found among the average relative abundance of archaeal genera between each dietary group. Moreover, the addition of linseed and hemp seed induced significant changes in FA concentration in the rumen, as a consequence of shift from C18 : 2n-6 to C18 : 3n-3 biohydrogenation pathway. Furthermore, dimethylacetal composition was affected by fat supplementation, as consequence of ruminal bacteria population modification. Finally, the association study between the rumen FA profile and the bacterial microbiome revealed that Fibrobacteriaceae is the bacterial family showing the highest and significant correlation with FA involved in the biohydrogenation pathway of C18 : 3n-3.     

Predicting duodenal flows and absorption of fatty acids from dietary characteristics in ovine and bovine species: a meta-analysis approach

Dietary and ruminal factors modify the ruminal biohydrogenation (RBH) of polyunsaturated fatty acids (FA), with duodenal FA flows being quantitatively and qualitatively different from FA intake. Using a meta-analysis approach from a database on duodenal flows of FA in ruminants, this study aimed to determine predictive equations for duodenal and absorbed flows of saturated FA, C18:1, C18:2 and C18:3 isomers, odd- and branched-chain FA (OBCFA), C20:5n-3, C22:5n-3 and C22:6n-3 and to quantify the effects of dietary and digestive factors on those equations. The database was divided into four subsets: forage, seed, vegetable oils or animal fats (oil/fat), and fish products (fish) subsets. Models of duodenal and absorbed FA flows were obtained through variance–covariance analysis. Effects of potential interfering factors (conservation mode and botanical families of forages, lipid source, technological processing of lipid supplements, diet composition and animal characteristics) were analysed. We obtained 83 models for duodenal FA flows as a function of FA intake for saturated FA (C14:0, C16:0 and C18:0), C18:1, C18:2 and C18:3 isomers and seven other models for OBCFA. For the seed/oil/fat subset, intakes of total C18:3, C18:2 and starch content increased the duodenal t11-C18:1 flow with 0.08, 0.16 and 0.005 g/kg of dry matter intake (DMI), respectively, whereas intake level [(DMI×100)/BW] decreased it. The c9c12c15-C18:3 RBH was higher for oil/fat than seed (96.7% v. 94.8%) and a protective effect of Leguminosae v. Gramineae against RBH for that FA appeared in the forage subset. The duodenal C17:0 flow increased with starch content and decreased with ruminal pH, respectively, whereas duodenal iso-C16:0 flow decreased with dietary NDF content for the seed/oil/fat subset. The duodenal C20:5n-3, C22:5n-3 and C22:6n-3 flows depended on their respective intake and the inhibitory effect of C22:6n-3 on duodenal C18:0 flow was quantified. Thirteen models of absorbed FA flows were performed depending on their respective duodenal flows. This study determined the effects of different qualitative and quantitative dietary and digestive factors, allowing for improved predictions of duodenal and absorbed FA flows.     

Rearing system and oleic acid supplementation effect on carcass and lipid characteristics of two muscles from an obese pig breed

Quality of pork depends on genotype, rearing and pre- and post-slaughter conditions. However, no information is available on rearing system changes and oleic acid supplementation on carcass characteristics and fatty acid (FA) profile of pork from the Alentejano (AL) pig, an obese breed. This study evaluates the effects of feeding low (LO) or high oleic acid diets (HO) to AL pigs reared in individual pens (IND) or outdoor (OUT) with access to pasture. Carcass composition was obtained and longissimus dorsi and semimembranosus samples were collected to analyse chemical composition and neutral and polar intramuscular lipids FA profile by gas chromatography. Statistical analysis was performed by a two-way ANOVA for rearing system and diet effects. OUT-reared pigs presented leaner carcasses than IND-reared ones. Both muscles presented lower intramuscular lipid content in OUT-reared pigs. Treatments affected the FA profile of muscles. Overall, OUT-reared pigs presented lower n-6/n-3 FA ratios, whereas pigs fed the HO diet exhibited lower saturated fatty acids (SFA), higher monounsaturated fatty acids (MUFA) levels and lower thrombogenic indexes on neutral intramuscular lipids than LO-fed pigs. On the polar fraction, OUT-reared pigs presented lower SAT and n-6/n-3 FA ratio, and higher polyunsaturated fatty acids (PUFA) levels on both muscles. Pigs fed the HO diet exhibited higher MUFA and lower PUFA levels on both muscles, and lower SAT levels on semimembranosus. This study shows rearing system and oleic acid supplementation have complementary effects and influence carcass composition and the nutritional quality of meat.     

Effects of resveratrol and genistein on growth,nutrient utilization and fatty acid composition of rainbow trout

The replacement of the finite and costly resource fish oil is an important task for aquaculture nutrition. A promising approach could be the use of plant bioactives that may have the potential to influence the metabolism and the synthesis of n-3 long chain polyunsaturated fatty acids, especially EPA (20:5n-3) and DHA (22:6n-3). In this study, the two phytochemicals resveratrol (RV) and genistein (G) were investigated for their effects on fish growth, nutrient utilization and body nutrient composition alongside their effects on whole body fatty acid (FA) composition. In a feeding trial lasting 8 weeks, rainbow trout (initial BW: 81.4±0.5 g) were held in a recirculating aquaculture system and fed six experimental diets with varying fish oil levels as plain variants or supplemented with 0.3% of dry matter (DM) of either RV or G. The six diets were as follows: diet F4 had 4% DM fish oil, diet F0 had 0% DM fish oil, diets F4+RV, F4+G, F0+RV and F0+G were equal to the diets F4 and F0, respectively, and supplemented with the phytochemicals RV and G. The feeding of the F0+RV diet resulted in reduced feed intake, growth rate and slightly reduced whole body lipid levels. At the same time, the amount of polyunsaturated FA and the n-3/n-6 ratio were significantly increased in whole body homogenates of rainbow trout fed diet F0+RV in comparison to the F0 control. The feeding of the F0+G diet led to reduced feed intake, slightly increased protein utilization but did not significantly affect the whole body FA composition. Overall, feeding the fish oil-free diet supplemented with the phytochemicals resulted in more pronounced effects on fish performance and FA composition than the single factors per se (dietary fish oil level or phytochemical). Present data indicate that G might not be of profitable use for trout nutrition. In terms of FA composition, RV could be a potentially useful complement for fish oil. However, the impairment of growth and performance parameters as observed in the present study discourages its use in trout diets.     

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.     

Effect of feeding lambs with a tanniferous shrub (rockrose) and a vegetable oil blend on fatty acid composition of meat lipids

The effects of feeding Cistus ladanifer (Cistus) and a blend of soybean and linseed oil (1 : 2 vol/vol) on fatty acid (FA) composition of lamb meat lipids and messenger RNA (mRNA) expression of desaturase enzymes was assessed. In total, 54 male lambs were randomly assigned to 18 pens and to nine diets, resulting from the combination of three inclusion levels of Cistus (50 v. 100 v. 200 g/kg of dry matter (DM)) and three inclusion levels of oil (0 v. 40 v. 80 g/kg of DM). The forage-to-concentrate ratio of the diets was 1 : 1. Longissimus muscle lipids were extracted, fractionated into neutral (NL) and polar lipid (PL) and FA methyl esters obtained and analyzed by GLC. The expression of genes encoding Δ5, Δ6 and Δ9 desaturases (fatty acid desaturase 1 (FADS1), fatty acid desaturase 2 (FADS2) and stearoyl CoA desaturase (SCD)) was determined. Intramuscular fat, NL and PL contents were not affected by oil or Cistus. Oil supplementation reduced (P<0.05) 16:0, c9-16:1, 17:0, c9-17:1 and c9-18:1 FA and increased (P<0.05) 18:2n-6, 18:3n-3 and the majority of biohydrogenation intermediates in NL. Cistus alone had few effects on FA of NL but interacted with oil (P<0.05) by increasing t10-18:1,t10,t12-18:2,t10,c12-18:2 and t7,c9-18:2. The t10-/t11-18:1 ratio increased with both Cistus and oil levels. The c9, t11-18:2 did not increase (P<0.05) with both oil and Cistus dietary inclusion. Oil reduced c9-16:1, 17:0, c9-17:1,c9-18:1, 20:4n-6, 22:4n-6 and 20:3n-9 proportions in PL, and increased 18:2n-6, 18:3n-3, 20:3n-3 and of most of the biohydrogenation intermediates. The Cistus had only minor effects on FA composition of PL. Cistus resulted in a reduction (P<0.05) of 20:5n-3 and 22:6n-3 in the meat PL. The expression level of SCD mRNA increased (P=0.015) with Cistus level, although a linear relationship with condensed tannins intake (P=0.11) could not be established. FADS1 mRNA expressed levels increased linearly (P=0.019) with condensed tannins intake. In summary, the inclusion of Cistus and oil in 1 : 1 forage-to-concentrate ratio diets resulted in a large increase in t10-18:1 and no increase in c9,t11-18:2 or n-3 long chain poor in polyunsaturated fatty acids in lamb meat.     

Fatty acid composition of membrane bilayers: Importance of diet polyunsaturated fat balance

In one of the most extensive analyses to date we show that the balance of diet n-3 and n-6 polyunsaturated fatty acids (PUFA) is the most important determinant of membrane composition in the rat under 'normal' conditions. Young adult male Sprague-Dawley rats were fed one of twelve moderate-fat diets (25% of total energy) for 8weeks. Diets differed only in fatty acid (FA) profiles, with saturate (SFA) content ranging 8-88% of total FAs, monounsaturate (MUFA) 6-65%, total PUFA 4-81%, n-6 PUFA 3-70% and n-3 PUFA 1-70%. Diet PUFA included only essential FAs 18:2n-6 and 18:3n-3. Balance between n-3 and n-6 PUFA is defined as the PUFA balance (n-3 PUFA as % of total PUFA) and ranged 1-86% in the diets. FA composition was measured for brain, heart, liver, skeletal muscle, erythrocytes and plasma phospholipids, as well as adipose tissue and plasma triglycerides. The conformer-regulator model was used (slope=1 indicates membrane composition completely conforming to diet). Extensive changes in diet SFA, MUFA and PUFA had minimal effect on membranes (average slopes 0.01, 0.07, 0.07 respectively), but considerable influence on adipose tissue and plasma triglycerides (average slopes 0.27, 0.53, 0.47 respectively). Diet balance between n-3 and n-6 PUFA had a biphasic influence on membrane composition. When n-3 PUFA<10% of total PUFA, membrane composition completely conformed to diet (average slope 0.95), while diet PUFA balance>10% had little influence (average slope 0.19). The modern human diet has an average PUFA balance ~10% and this will likely have significant health implications.     

Lipid classes and fatty acid composition of rotifers (Brachionus plicatilis) fed two algal diets

Rotifers (Brachionus plicatilis), maintained on baker's yeast, were fed for 24h upon two algal diets, Isochrysis galbana (diet A) and Isochrysis galbana + Nannochloropsis gaditana (diet B). (These algal diets were selected for their potential use as essential fatty acid (EFA) boosters, taking into account the requirements of fish larvae). The effect of these algal diets on total lipid content, lipid classes and fatty acid composition was studied. The total lipid content increased after feeding upon both diets but no significant differences were found between the two types. Neutral lipid and polar lipid contents increased and a positive correlation was observed between the neutral lipids content of rotifers and that of the food supplied. However, the content of polar lipids in rotifers did not depend upon that of the diet. The increase in neutral lipid content was found to be higher in rotifers fed upon diet B, compared to diet A which increased the phospholipid content. Non-enriched rotifers contained only small amounts of polyenoic fatty acids, i.e. 18:3n-6, 18:3n-3, 20:4n-6, 20:5n-3 and 22:6n-3, the contents of which increased significantly by feeding both diets. The EFA composition (20:4n-6, 20:5n-3 and 22:6n-3) of neutral lipids and phopholipids in rotifers reflected the EFA composition of each diet. Diet B-fed rotifers had the highest content in 20:4n-6 and 20:5n-3, whereas rotifers fed diet A and the highest 22:6n-3 content. The mixed diet I. galbana + N. gaditana enhanced substantially the composition of lipid classes i.e. neutral lipids and of n-3 PUFA of rotifers in comparison with Isochrysis or yeast diets.     

Dietary fish oil and flaxseed for rabbit does: fatty acids distribution and Δ6-desaturase enzyme expression of different tissues

Standard feeds are imbalanced in term of n-6/n-3 polyunsaturated fatty acids (PUFA) ratio, with a low proportion of the latter. The reproductive system appears to be strongly affected by administration of n-3 PUFA, and ingredients rich in α-linolenic acid (ALA; i.e. vegetable sources) or EPA and DHA acids (i.e. fish oil) can be included in animal diets to balance PUFA intake. The aim of this study was to evaluate the effect of dietary supplementation with flaxseed (ALA) or fish oil (EPA and DHA) on PUFA metabolism in rabbit does. A total of 60 New Zealand White female rabbits were assigned to three experimental groups: control group, FLAX group fed 10% extruded flaxseed and FISH group fed 3% fish oil. Blood, milk, liver and ovaries were collected from the does to assess the lipid composition; furthermore, FADS2 gene expression was assessed in liver and ovary tissues. Reproductive performance of does was also recorded. The fertility rate and number of weaned rabbits improved with n-3 dietary supplementation: does at first parity showed the lowest reproductive results, but the administration of n-3 reduced the gap between primiparous and multiparous does. Feed consumption and milk production were not affected by the feeding regime. The fatty acid composition of milk, plasma, liver and ovaries were widely influenced by diet, showing higher concentrations of n-3 long-chain PUFA (LCP) in does fed with n-3 enriched diets. FISH diet resulted in the highest n-3 LCP enrichment, whereas in the FLAX group, this increase was lower. Blood and milk showed low levels of LCP, whereas liver and ovaries were the main sites of n-3 LCP synthesis and accumulation. Accordingly, although the liver is the main metabolic centre for LCP synthesis, ovaries also have a prominent role in LCP generation. FADS2 expression in liver and ovary tissue was downregulated by FISH administration. In conclusion, the enrichment of diets with n-3 PUFA could be an effective strategy for improving the reproductive performance of does.     

Evolution of the fatty acid profile of subcutaneous back-fat adipose tissue in growing Iberian and Landrace × Large White pigs

The lipid content and fatty acid (FA) profile in pig tissues are strongly influenced by genotype and nutrient supply, with implications in meat quality. The de novo lipid synthesis and pattern of FA unsaturation could be an important cause of variation in the overall efficiency of energy utilization among breeds. To test the effects of pig genotype and CP supply on the evolution of back-fat tissue FA profile throughout the growing and finishing stages, 32 Iberian (IB) and Landrace × Large White (LR × LW) barrows were offered one of two diets differing in CP content (13% or 17% as fed). A pair-fed procedure (0.8 × ad libitum intake of IB pigs) was used. Subcutaneous fat samples were taken at the dorso-lumbar region at ∼38, 50, 65, 90 and 115 kg BW. Higher proportions of total monounsaturated FA (MUFA; P < 0.01) and lower proportions of total saturated FA (SFA; P < 0.01 to 0.05) were found in the outer back-fat layer of pigs both at 50 and 115 kg BW. Pig genotype affected the FA composition of both subcutaneous back-fat layers. The proportions of C18:0 and SFA in fat tissue were higher in IB than in LR × LW pigs from 38 to 65 kg BW, especially in the outer layer. In addition, MUFA contents were higher in IB pigs at 115 kg BW in both layers (+5% on average; P < 0.01). Increased proportions of C18:2 n-6 and polyunsaturated FA (PUFA) were found in LR × LW pigs, irrespective of the stage of growth and back-fat layer (P⩽0.02). At 50 kg BW, pigs receiving the high-protein diet presented the highest C18:2 n-6, C18:3 n-3, C20:5 n-3 and PUFA contents. A significant genotype × CP content interaction was observed for C18:3 n-3 because of the increased concentration of this FA in LR × LW pigs when offered the 17% CP diet (P < 0.05). Higher C16:0 and SFA contents (+5%; P = 0.03) were found in pigs offered the 13% CP diet and slaughtered at 115 kg BW. There was a genotype × CP interaction for MUFA concentration because of the higher MUFA content observed in IB pigs offered the highest protein content diet (P = 0.03). Our results suggest that genetic variation in de novo lipid synthesis and pattern of FA unsaturation might contribute to explain differences in back-fat FA profile of IB and LR × LW pigs under identical nutritional management. They could be also relevant to explain the low efficiency of nutrient and energy utilization in the IB pig.     

Changes in fatty acid composition of Mytilus galloprovincialis (Lmk) fed on microalgal and wheat germ diets

Dietary fatty acid incorporation and changes in various lipid and phospholipid classes in the mussel Mytilus galloprovincialis subjected to three different dietary regimens were analysed and compared. Group A was unfed; group B received a diet consisting of 100% Thalassiosira weissflogii, exhibiting the typical fatty acid composition of diatoms, and group C received a diet consisting of 100% wheat germ conferring a 18:2:n-6 abundance. Biochemical analyses of diets and mussels were carried out at the beginning and at the end of the 30-day experimental period. Starvation and T. weissflogii based diet poorly affected mussel growth and fatty acid composition which remained unchanged. On the contrary, the wheat germ-based diet increased the condition index and deeply affected the fatty acid profile of all lipid and phospholipid classes. The high dietary 18:2n-6 level drastically reduced tissue content of 20:4n-6, 20:5n-3 and 22:6n-3. The biosynthesis of Non Methylene Interrupted (NMI) dienoic fatty acid appeared to be insensitive to the high input of 16:1n-7 and 18:1n-9 respectively from diet B and C, and to the PUFA shortage of diet C. Nevertheless the two NMI trienoic derivatives, 20:3Δ5,11,14 and 22:3Δ7,13 16, were found higher in C with respect to other groups, presumably due to the high 18:2n-6 content of this diet.     

Dietary eicosapentaenoic acid supplementation accentuates hepatic triglyceride accumulation in mice with impaired fatty acid oxidation capacity

Reduced mitochondrial fatty acid (FA) β-oxidation can cause accumulation of triglyceride in liver, while intake of eicosapentaenoic acid (EPA) has been recommended as a promising novel therapy to decrease hepatic triglyceride content. However, reduced mitochondrial FA β-oxidation also facilitates accumulation of EPA. To investigate the interplay between EPA administration, mitochondrial activity and hepatic triglyceride accumulation, we investigated the effects of EPA administration to carnitine-deficient mice with impaired mitochondrial FA β-oxidation. C57BL/6J mice received a high-fat diet supplemented or not with 3% EPA in the presence or absence of 500 mg mildronate/kg/day for 10 days. Liver mitochondrial and peroxisomal oxidation, lipid classes and FA composition were determined. Histological staining was performed and mRNA level of genes related to lipid metabolism and inflammation in liver and adipose tissue was determined. Levels of pro-inflammatory eicosanoids and cytokines were measured in plasma. The results showed that mildronate treatment decreased hepatic carnitine concentration and mitochondrial FA β-oxidation and induced severe triglyceride accumulation accompanied by elevated systemic inflammation. Surprisingly, inclusion of EPA in the diet exacerbated the mildronate-induced triglyceride accumulation. This was accompanied by a considerable increase of EPA accumulation while decreased total n-3/n-6 ratio in liver. However, inclusion of EPA in the diet attenuated the mildronate-induced mRNA expression of inflammatory genes in adipose tissue. Taken together, dietary supplementation with EPA exacerbated the triglyceride accumulation induced by impaired mitochondrial FA β-oxidation. Thus, further thorough evaluation of the potential risk of EPA supplementation as a therapy for NAFLD associated with impaired mitochondrial FA oxidation is warranted.