Evaluation of two GC columns (60-m SUPELCOWAX 10 and 100-m CP Sil 88) for analysis of milkfat with emphasis on CLA, 18:1, 18:2 and 18:3 isomers,and short- and long-chain FA

Milkfat is a complex mixture of many diverse FA, some of which have demonstrated health benefits including anticancer properties. Attempts are under way to enrich milkfats with long-chain n−3 PUFA and CLA. It has been recommended that the analysis of these milkfats requires gas chromatography (GC) equipped with long, highly polar capillary columns. However, many analyses have been reported using CARBOWAX^TM type (polyethylene glycol) capillary columns, such as SUPELCOWAX 10, even though the separation characteristics of many of the FA and their isomers present in milkfats have not been described in detail. This includes the isomers of CLA, cis- and trans-octadecenoic acid (18∶1), linoleic acid (18∶2n−6), and linolenic acid (18∶3n−3), and the long-chain PUFA. On the other hand, the resolution of these FA and their isomers has been more fully described using the highly polar capillary columns, such as CP Sil 88 and SP2560 because of the improved resolution obtained using these polar columns. The present study was undertaken to characterize the separation of these FA present in milkfats using a 60-m SUPELCOWAX 10 column, to compare the results to those from a 100-m CP Sil 88 column, and to determine if these two columns could possibly serve to complement each other for the analysis of total milkfat. The advantages of the SUPELCOWAX 10 column were a better resolution of the short-chain saturated from their monounsaturated FA (MUFA) analogs, and a complete separation of the α-linolenic (18∶3n−3) and eicosadecenoic acid (20∶1) isomers. It also provided an alternative elution order of the linoleic (18∶2n−6), 18∶3n−3 and γ-linolenic (18∶3n−6) acid isomers. On the other hand, the CP Sil 88 column provided a better resolution of the CLA isomers, MUFA, the isolated cis and trans MUFA fractions, the PUFA, and many the 18∶2n−6 and 18∶3n−3 isomers. A complete analysis of milk lipids using the CP Sil 88 column required the prior separation of total FAME using silver ion-TLC. The results of the present study confirm that the 100-m highly polar capillary GC columns are mandatory for the analysis of milk lipids, and at best, the 60 m SUPELCOWAX 10 capillary column serves as a complementary GC column to provide different separations in certain regions based on its intermediate polarity.     

Structural importance of thecis-5 ethylenic bond in the endogenous desaturation product of dietary elaidic acid,cis-5,trans-9 18∶2 acid,for the acylation of rat mitochondria phosphatidylinositol

When rats were fed elaidic (trans-9 18∶1) acid at a high load in diets that were otherwise marginally or almost completely deficient in linoleic (cis-9,cis-12 18∶2) acid, elaidic acid was desaturated tocis-5,trans-9 18∶2 acid. This polymethylene-interrupted acid was then incorporated into most phospholipids from rat mitochondria, cardiolipin being an exception. Its level of esterification in phospholipids followed the increasing order: phosphatidylethanolamine <phosphatidylcholine < phosphatidylinositol (PI). The content ofci-5,trans-9 18∶2 acid decreased in organs in the order liver > kidney > heart. The levels ofcis-5,trans-9 18∶2 acid increased in mitochondria phospholipids as the level of linoleic acid was lowered in the diet. In liver mitochondria PI, it reached 16% of total fatty acids. After hydrolysis of liver mitochondria PI withNaja naja phospholipase A₂, we observed that elaidic acid was essentially esterified to position 1 at the expense of saturated acids, whereascis-5,trans-9 18∶2 acid was exclusively esterified to position 2, along with 20∶3n−9 and 20∶4n−6 acids. As a consequence, the sums of saturated andtrans-9 18∶1 acids on the one hand, and of 20∶3n−9, 20∶4n−6, andcis-5,trans-9 18⩺2 acids on the other hand, remained fairly constant in liver mitochondria PI (ca. 55 and 30%, respectively). Becausetrans-9 18∶1 andcis-5,trans-9 18∶2 acids differ only by thecis-5 ethylenic bond, which is also present in 20∶3n−9 and 20∶4n−6 acids, this distribution pattern indicates that thecis-5 double bond, rather than any other ethylenic bond, may be of major structural importance for channeling fatty acids to position 2 of PI.     

Dietary linoleic acid and the fatty acid profiles in rats fed partially hydrogenated marine oils

The influence of the linoleic acid levels of diets containing partially hydrogenated marine, oils (HMO) rich in isomeric 16∶1, 18∶1, 20∶1 and 22∶1 fatty acids on the fatty acid profiles of lipids from rat liver, heart and adipose tissue was examined. Five groups of rats were fed diets containing 20 wt% fat−16% HMO+4% vegetable oils. In these diets, the linoleic acid contents varied between 1.9% and 14.5% of the dietary fatty acids, whereas the contents oftrans fatty acids were 33% in all groups. A sixth group was fed a partially hydrogenated soybean oil (HSOY) diet containing 8% linoleic acid plus 32%trans fatty acids, mainly 18∶1, and a seventh group, 20% palm oil (PALM), with 10% linoleic acid and notrans fatty acids. As the level of linoleic acid in the HMO diets increased from 1.9% to 8.2%, the contents of (n−6) polyunsaturated fatty acids (PUFA) in the phospholipids increased correspondingly. At this dietary level of linoleic acid, a plateau in (n−6) PUFA was reached that was not affected by further increase in dietary 18∶2(n−6) up to 14.5%. Compared with the HSOY- or PALM-fed rats, the plateau value of 20∶4(n−6) were considerably lower and the contents of 18∶2(n−6) higher in liver phosphatidylcholines (PC) and heart PC. Heart phosphatidylethanolamines (PE) on the contrary, had elevated contents of 20∶4(n−6), but decreased 22∶5(n−6) compared with the PALM group. All groups fed HMO had similar contents oftrans fatty acids, mainly 16∶1 and 18∶1, in their phospholipids, irrespective of the dietary 18∶2 levels, and these contents were lower than in the HSOY group. High levels of linoleic acid consistently found in triglycerides of liver, heart and adipose tissue of rats fed HMO indicated that feeding HMO resulted in a reduction of the conversion of linoleic acid into long chain PUFA that could not be overcome by increasing the dietary level of linoleic acid.     

Hyperphagia modifies FA profiles of plasma phospholipids,plasma FFA,and adipose tissue TAG

Hyperphagia was achieved by continuous intracerebroventricular infusion of a melanocortin receptor antagonist (HS024; Neosystem, Strasbourg, France) in rats. The effects of hyperphagia on FA composition and concentration of plasma phospholipids (PL), plasma FFA, and adipose tissue TAG were studied in rats for 8 d [short-term hyperphagia (STH); n=8], or 28 d [longterm hyperphagia (LTH); n=9]. The control rats were treated with artificial cerebrospinal fluid for 8 d (n=8) or 28 d (n=10). The rats were fed the same regular diet. In STH rats the plasma PL and fasting plasma FFA contained higher concentrations of saturated FA (SFA) and monounsaturated FA (MUFA), and plasma FFA contained lower n−6 PUFA than in the control rats. In LTH rats the plasma PL contained higher concentrations of SFA, MUFA, and n−3 PUFA and higher proportions of 16∶1n−7 and 18∶1n−9 at the expense of 18∶2n−6 than in the control rats. In LTH rats the abundant dietary intake of 18∶2n−6 did not enrich 18∶2n−6 of the plasma PL or adipose tissue TAG. In LTH rats the fasting plasma FFA contained more than twofold higher concentrations of SFA and MUFA, and higher proportions of 16∶1n−7 and 18∶1n−9 at the expense of 18∶2n−6 than in the control rats. This animal obesity model shows that LTH affects the FA composition and concentration of plasma PL, plasma FFA, and adipose tissue TAG, a result consistent with changes associated with increased risk of various diseases in humans. These results also demonstrate that LTH alters the FA composition of plasma PL and adipose tissue TAG in a way that does not reflect the FA composition of dietary fat.     

Comparison of the effects of dietary alpha-linolenic,stearidonic, and eicosapentaenoic acids on production of inflammatory mediators in mice

The effects of dietary stearidonic acid (18∶4n−3) on inflammatory mediator release in whole blood and splenocytes was investigated in Balb/c mice, and the effects were compared with those of two other n−3 PUFA: α-linolenic acid (18∶3n−3) and EPA (20∶5n−3). TAG mixtures containing 10% of 18∶4n−3, 18∶3n−3, or 20∶5n−3 as the respective sole n−3 PUFA were enzymatically synthesized. Diets containing synthesized TAG mixtures were fed to Balb/c mice for 3 wk. The release of prostaglandin E₂ (PGE₂) and tumor necrosis factor (TNF) were measured in whole blood and splenocytes stimulated with lipopolysaccharide. In whole blood, the production of INF was suppressed by all dietary n−3 PUFA (18∶3n−3, 18∶4n−3, and 20∶5n−3) as compared with the control diet, which contained TAG prepared from safflower oil. PGE₂ production was not significantly changed. Differences among the n−3 PUFA (18∶3n−3), 18∶4n−3, and 20∶5n−3) were not observed. In splenocytes, PGE₂ production was suppressed by dietary n−3 PUFA, but TNF production was not. GC analysis of plasma and splenocyte FA profiles showed an increase in the levels of 20∶4n−3, 20∶5n−3, and 22∶6n−3 in mice fed the diet containing 18∶4n−3.     

Incorporation ofcis- andtrans-octadecenoic acids into the membranes of rat liver mitochondria

The incorporation of dietary isomeric fatty acids into the membranes of liver mitochondria was investigated. Three groups of rats were fed diets containing 3% sunflower seed oil plus 15%, 20%, or 25% partially hydrogenated arachis oil. A fourth group was fed 25% partially hydrogenated arachis oil, but no sunflower seed oil. All diets were given for 3, 6, or 10 weeks. After 10 weeks, the content oftrans fatty acids in the lipids of the mitochondrial membranes was 15–19% of the total fatty acids. The composition of thetrans- and thecis-octadecenoic acids in the lipids of the mitochondrial membranes was similar for all groups supplemented with sunflower seed oil (SO), irrespective of time and dietary level of partially hydrogenated arachis oil (HAO). Thecis 18∶1 (n−8), which was a major isomer of the partially hydrogenated arachis oil, was almost excluded from the mitochondrial fatty acids. Likewise, the content oftrans 18∶1 (n−8) was considerably lower in the mitochondrial lipids than in the diet. On the contrary, the content oftrans 18∶1 (n−6) was higher in the mitochondrial lipids than in the diet. In the group fed without sunflower seed oil, isomers of linoleic acid and arachidonic acid were observed in the lipids of mitochondrial membranes. Presented in part at the ISF Congress, Marseille, September 1976.     

Incorporation of n−3 fatty acids into plasma and liver lipids of rats: Importance of background dietary fat

The health benefits of long-chain n−3 PUFA (20∶5n−3 and 22∶6n−3) depend on the extent of incorporation of these FA into plasma and tissue lipids. This study aimed to investigate the effect of the background dietary fat (saturated, monounsaturated, or n−6 polyunsaturated) on the quantitative incorporation of dietary 18∶3n−3 and its elongated and desaturated products into the plasma and the liver lipids of rats. Female weanling Wistar rats (n=54) were randomly assigned to six diet groups (n=9). The fat added to the semipurified diets was tallow (SFA), tallow plus linseed oil (SFA-LNA), sunola oil (MUFA), sunola oil plus linseed oil (MUFA-LNA), sunflower oil (PUFA), or sunflower oil plus linseed oil (PUFA-LNA). At the completion of the 4-wk feeding period, quantitative FA analysis of the liver and plasma was undertaken by GC. The inclusion of linseed oil in the rat diets increased the level of 18∶3n−3, 20∶5n−3, and, to a smaller degree, 22∶6n−3 in plasma and liver lipids regardless of the background dietary fat. The extent of incorporation of 18∶3n−3, 20∶5n−3, and 22∶5n−3 followed the order SFA-LNA>MUFA-LNA>PUFA-LNA. Levels of 22∶6n−3 were increased to a similar extent regardless of the type of major fat in the rat diets. This indicates that the background diet affects the incorporation in liver and plasma FA pools of the n−3 PUFA with the exception of 22∶6n−3 and therefore the background diet has the potential to influence the already established health benefits of long-chain n−3 fatty acids.     

Fatty acids bound to <Emphasis Type="Italic">Fasciola hepatica</Emphasis> 12 kDa fatty acid-binding protein,a candidate vaccine,differ from fatty acids in extracts of adult flukes

The FA composition of Fasciola hepatica 12 kDA purified native FA-binding protein (nFh12), a candidate vaccine against fascioliasis, is described. The FA chain lengths ranged between 12 and 24 carbons. The principal FA were 16∶0 18∶1n−9, 18∶0, 20∶4n−6, and 20∶1n−9. The acids 16∶0, 18∶1n−9, and 18∶0 comprised over half the FA that were bound to the whole FA-binding protein. Small amounts (1.0–2.8%) of isoanteiso methyl-branched FA also were characterized. Forty-one different FA were identified in extracts of the adult flukes, with the three most abundant FA also being 16∶0, 18∶1n−9, and 18∶0. A similar proportion of saturated vs. unsaturated FA was observed between the whole extract from F. hepatica and the nFh12 protein. However, the n−3/n−6 ratio of PUFA was significantly different, being 1.2 in the whole extract vs. 9.6 in the nFh12 protein complex. The nFh12 protein binds more n−5, n−6, and n−7 PUFA and less n−3 and n−9 PUFA than the whole extract. In addition, cholesterol (56%), sitosterol (36%), and fucosterol (8%) also were bound to the nFh12 protein complex.     

The effect of conjugated linoleic acid isomers on fatty acid profiles of liver and adipose tissues and their conversion to isomers of 16:2 and 18:3 conjugated fatty acids in rats

Conjugated linoleic acid (CLA) is a collective term that describes different isomers of linoleic acid with conjugated double bonds. Although the main dietary isomer is 9cis,11trans-18∶2, which is present in dairy products and ruminant fat, the biological effects of CLA generally have been studied using mixtures in which the 9cis,11trans- and the 10trans,12cis-18∶2 were present at similar levels. In the present work, we have studied the impact of each isomer (9cis,11trans- and 10trans,12cis-18∶2) given separately in the diet of rats for 6 wk. The 10trans,12cis-18∶2 decreased the triacylglycerol content of the liver (−32%) and increased the 18∶0 content at the expense of 18∶1n−9, suggesting an alteration of the Δ9 desaturase activity, as was already demonstrated in vitro. This was not observed when the 9cis,11trans-18∶2 was given in the diet. Moreover, the 10trans,12cis-18∶2 induced an increase in the C₂₂ polyunsaturated fatty acids in the liver lipids. The 10trans,12cis-18∶2 was mainly metabolized into conjugated 16∶2 and 18∶3, which have been identified. The 9cis,11trans isomer was preferentially metabolized into a conjugated 20∶3 isomer. Thus, the 9cis,11trans- and the 10trans,12cis-CLA isomers are metabolized differently and have distinct effects on the metabolism of polyunsaturated fatty acids in rat liver while altering liver triglyceride levels differentially.     

Identification of noveltrans isomers of 20∶5n−3 in liver lipids of rats fed a heated oil

Trans polyunsaturated n−3 fatty acids are formed as a result of the heat treatment of vegetable oils. It was demonstrated previously that the 18∶3 Δ9cis, 12cis, 15trans containing acis Δ9 ethylenic bond was converted to a geometrical isomer of 20∶5n−3, the 20∶5 Δ5cis, 8cis, 11cis, 14cis, 17trans. In the present study, we have identified two new isomers of eicosapentaenoic acid, the Δ11 monotrans and the Δ11, 17 ditrans isomers in liver of rats fed a heated oil. These are formed as a result of the conversion of two of the main isomers of linolenic acid which are present in refined and frying oils, the 18∶3 Δ9trans, 12cis, 15cis and the 18∶3 Δ9trans, 12cis, 15trans.     

The Effects of dietary n−3/n−6 ratio on brain development in the mouse: a dose response study with long-chain n−3 fatty acids

This study examines the effects of the ratio of n−3/n−6 fatty acids (FA) on brain development in mice when longchain n−3 FA are supplied in the diet. From conception until 12 days after birth, B6D2F₁ mice were fed liquid diets, each providing 10% of energy from olive oil, and a further 10% from different combinations of free FA concentrates derived from safflower oil (18∶2n−6), and fish oil (20∶5n−3 and 22∶6n−3). The range of dietary n−3/n−6 ratios was 0,025, 0.5, 1.0, 2.0, and 4.0, with an n−6 content of greater than 1.5% of energy in all diets, and similar levels of total polyunsaturated fatty acids (PUFA). In an additional group of ratio 0.5, 18∶2n−6 was partially replaced by its δ6 desaturation product, 18∶3n−6. Biochemical analyses were conducted on 12-day-old pup brains, as well as on samples of maternal milk. No obvious effects on overall pup growth and development were observed, apart from a smaller litter size at ratio 1. Co-variance analysis indicated that increasing the n−3/n−6 ratio was associated with slightly smaller brains, relative to body weight. We found that 18∶2n−6 and 20∶5n−3 were the predominant n−6 and n−3 FA in the milk; in the brain these were 20∶4n−6 and 22∶6n−3, respectively. Increasing dietary n−3/n−6 ratios generally resulted in an increase in n−3 FA, with a corresponding decrease in n−6 FA. The n−3/n−6 ratio of the milk lipids showed a strong linear relationship with the diet, but in the brain the rate of increase tended to decrease beyond 0.5 (phosphatidylcholine, PC) and 0.25 (phosphatidylethanolamine, PE), such that there was a significant quadratic contribution to the relationship. The partial replacement of dietary 18∶2n−6 with 18∶3n−6 raised levels of 20∶4n−6 in milk, brain PC, and brain PE. These results indicate that the n−3/n−6 ratio of the phospholipids in the developing mouse brain responds maximally to maternal dietary long-chain n−3/n−6 ratios of between 0.25 and 0.5.     

Dietary fats containing concentrates ofcis ortrans octadecenoates and the patterns of polyunsaturated fatty acids of liver phosphatidylcholine and phosphatidylethanolamine

The effects of the mixedcis- 18∶1 isomers and mixedtrans-18∶1 isomers present in partially hydrogenated soybean oil (PHSO) upon the patterns of polyunsaturated fatty acids (PUFA) in liver phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were studied in rats fed concentrates ofcis- 18∶1 ortrans- 18∶1 isomers isolated as triacylglycerides from PHSO. Thecis- 18∶1 andtrans- 18∶1 concentrates were fed at levels equal to those present in PHSO fed at 17.9% of the diet. All diets contained the required amounts of both linoleic and linolenic acids. Thetrans- 18∶1 concentrate was found to suppress the levels of 20∶4ω6 and 20∶3ω9, and to increase the levels of 18∶2ω6 and 20∶5ω3 in PC and PE. Thecis- 18∶1 concentrate suppressed 20∶4ω6 in PC, 20∶5ω3 in PC and PE, and 18∶2ω6 was more effective than thetrans concentrate in suppressing 22∶6ω3. Thetrans- 18∶1 concentrate was more effective in suppressing 20∶4ω6. Thetrans-18∶ isomers appear to modify PUFA metabolism by inhibition of PUFA synthesis, whereas thecis- 18∶1isomers appear to compete with 2-position fatty acyl transfer and to inhibit ω3 PUFA acylation.     

Effect of pasture vs. concentrate diet on CLA isomer distribution in different tissue lipids of beef cattle

This study examined the effects of feeding pasture vs. concentrate on the distribution of CLA isomers in the lipids of longissimus and semitendinosus muscle, liver and heart muscle, and subcutaneous fat in beef bulls. Sixty-four German Holstein and German Simmental bulls were randomly allocated to either an indoor concentrate system or periods of pasture feeding followed by a finishing period on a concentrate containing linseed to enhance their beef content of n−3 PUFA and CLA. The concentrations of CLA isomers in the different tissues were determined by GC and silver ion HPLC. The diet affected the distribution of individual CLA isomers in the lipids of the different tissues. The concentration (mg/100 g fresh tissue) of the most prominent isomer, cis-9,trans-11 18∶2, was increased up to 1.5 times in liver and heart tissue of bulls fed on pasture as compared with concentrate. However, no diet effect was observed for cis-9,trans-11 18∶2 in the lipids of longissimus muscle and subcutaneous fat. In all tissues, the second-most abundant CLA isomer in concentratefed bulls was trans-7,cis-9 18∶2. In contrast, trans-11,cis-13 18∶2 was the second-most abundant CLA isomer in all investigated tissue lipids of pasture-fed bulls. The concentration of the trans-11,cis-13 18∶2 isomer was up to 15 times higher in tissues of pasture-fed bulls as compared with concentrate-fed animals. Furthermone, diet affected the concentrations of the CLA trans,trans 18∶2 isomers. Pasture feeding significantly increased the concentrations of some trans,trans 18∶2 isomers as compared with concentrate, predominantly trans-12,trans-14 18∶2 and trans-11,trans-13 18∶2. Overall, pasture feeding resulted in significantly increased concentrations of the sum of CLA isomers in the lipids of longissimus, muscle, subcutaneous fat, heart and liver muscle of German Holstein and German Simmental bulls, but not in semitendinosus muscle.     

Incorporation oftrans long-chain n−3 polyunsaturated fatty acids in rat brain structures and retina

During heat treatment, polyunsaturated fatty acids and specifically 18∶3n−3 can undergo geometrical isomerization. In rat tissues, 18∶3 Δ9c, 12c, 15t, one of thetrans isomers of linolenic acid, can be desaturated and elongated to givetrans isomers of eicosapentaenoic and docosahexaenoic acids. The present study was undertaken to determine whether such compounds are incorporated into brain structures that are rich in n−3 long-chain polyunsaturated fatty acids. Two fractions enriched intrans isomers of α-linolenic acid were prepared and fed to female adult rats during gestation and lactation. The pups were killed at weaning. Synaptosomes, brain microvessees and retina were shown to contain the highest levels (about 0.5% of total fatty acids) of thetrans isomer of docosahexaenoic acid (22∶6 Δ4c, 7c, 10c, 13c, 16c, 19t). This compound was also observed in myelin and sciatic nerve, but to a lesser extent (0.1% of total fatty acids). However, the ratios of 22∶6trans to 22∶6cis were similar in all the tissues studied. When the diet was deficient in α-linolenic acid, the incorporation oftrans isomers was apparently doubled. However, comparison of the ratios oftrans 18∶3n−3 tocis 18∶3n−3 in the diet revealed that thecis n−3 fatty acids were more easily desaturated and elongated to 22∶6n−3 than the correspondingtrans n−3 fatty acids. An increase in 22∶5n−6 was thus observed, as has previously been described in n−3 fatty acid deficiency. These results encourage further studies to determine whether or not incorporations of suchtrans isomers into tissues may have physiological implications. Presented in part at the 32nd International Conference on the Biochemistry of Lipids, 1991, Granada, Spain. Delta nomenclature (Δ) is used fortrans polyunsaturated fatty acids to specify the position and geometry of ethylenic bonds. Polyunsaturated fatty acids containingtrans double bonds are abbreviated giving the locations of thetrans double bonds only; e.g., 20∶5 Δ17t 20∶5 Δ5c,8c,11c,14c,17t; 22∶5 Δ19t, 22∶5 Δ7c,10c,13c,16c,19t; 22∶6 Δ19t 22∶6 Δ4c,7c,10c,13c,16c,19t.     

Lipid class and fatty acid composition of the boreal soft coral Gersemia rubiformis

Total lipid, phospholipid, and FA composition and distribution of FA between polar lipids (PL) and neutral lipids (NL) were investigated in the boreal soft coral Gersemia rubiformis from the Bering Sea. The total lipids were mostly hydrocarbons and waxes (33.7%) and PL (33.1%). The content of monoalkyldiacylglycerols (9.7%) exceeded the content of TAG (6.7%). PC and PE constituted 31.4% and 25.6% of total phospholipids, respectively. Principal FA were 16∶0, 16∶1n−7, 18∶0, 18∶1n−9, 18∶1n−7, 20∶1n−7, 20∶4n−6, 20∶4n−3, 20∶5n−3 22∶5n−3, 22∶6n−3, 24∶5n−6, and 24∶6n−3. Most n−6 PUFA (52% of total FA) were associated with the PL fraction; this was especially true for arachidonic and tetracosapentaenoic acids. The NL were enriched with mono-, di-, trienoic, and n−3 PUFA. The variation in EPA levels in both NL and PL suggests an origin of this acid from lipids of diatoms consumed by the corals.     

Maternal dietary FA modulate the long-chain n−3 PUFA status of chick cardiac tissue

The effect that egg yolk or maternal n−3 FA have on the cardiac tissue long-chain n−3 FA status of chicks during growth was investigated. Fggs with low, medium, and high levels of n−3 PUFA were obtained by feeding breeder hens a wheat/soybean meal-based diet containing 5% sunflower oil (Low n−3), 2.5% sunflower oil plus 2.5% fish oil (Medium n−3), or 5% fish oil (High n−3). The chicks hatched from Low, Medium, and High n−3 eggs were fed a diet containing 18∶3n−3, but devoid of long-chain n−3 FA. The FA composition of cardiac tissue was determined on days 0, 14, 28, and 42. At day 0, the cardiac FA reflected maternal diet. With time, the level of all the long-chain n−3 FA decreased compared with day 0, and this was true especially by day 14. These data show that dietary 18∶3n−3 fed to the chicks did not sustain high levels of EPA and DHA in cardiac tissue, despite the high content of long-chain n−3 FA in the maternal diet. At days 0 and 14, the chicks hatched from High and Medium n−3 eggs had higher 20∶5n−3, 22∶5n−3, and 22∶6n−3 contents with a concomitant reduction in 20∶4n−6 in the cardiac tissue compared with the Low n−3 egg group. Cardiac tissue of birds hatched from Medium n−3 eggs retained higher levels of 20∶5n−3 up to day 42 of growth when compared with other treatments (P<0.05). None of the treatments was effective in maintaining DHA levels after day 14 of growth.     

Trans fatty acids. 2. Fatty acid composition of the brain and other organs in the mature female pig

Female pigs were fed from three wk of age and up to two years a diet containing partially hydrogenated fish oil (PHFO, 28%trans monoenoic fatty acids), partially hydrogenated soybean oils (PHSBO, 36%trans fatty acids) or lard. No consistent differences were found between PHFO and PHSBO with regard to incorporation oftrans fatty acids in organ lipids, buttrans incorporations were highly organ-specific. Notrans fatty acids were detected in brain phosphatidylethanolamine (PE). The incorporation of monoenoictrans isomers, as a percentage of totalcis + trans, in other organs was highest in subcutaneous adipose tissue and liver mitochondria PE, followed by blood lipids with the lowest level in heart PE. The percentage oftrans isomers compared with that of dietary lipids was consistently lower for 20∶1, compared with 18∶1 in organs from PHFO-fed pigs. The only effect of dietarytrans fatty acids on the fatty acid pattern of brain PE was an increased level of 22∶5n−6. Heart PE and total serum lipids of pigs fed the hydrogenated fats contained higher levels of 18∶2n−6, and these lipids of the PHFO-fed group also contained slightly elevated amounts of 20∶3n−6, 18∶3n−3 and 20∶5n−3. Liver mitochondria PE of the PHFO group also contained higher levels of 20∶3n−6 and 22∶5n−6. Dietarytrans fatty acids caused a consistent decrease of saturated fatty acids compensated by increased levels of monoenes. Thus, it may be concluded that dietary long-chaintrans fatty acids in PHFO behaved similarly metabolically to 18∶1-trans in PHSBO in pigs, without noticeable influence on brain PE composition and with moderate to slight effects on the fatty acid profile of the other organs.     

Lipid and FA composition of the pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis>: Influence of season and maturation

The lipid and FA composition of the total lipids of the pearl oyster Pinctada fucata martensii, in different seasons and in different areas, were analyzed to clarify its lipid physiology and to estimate the possible influence of its prey phytoplankton. TAG and sterols were the major components in the neutral lipids in all conditions, whereas high levels of phospholipids (PE and PC) were found in the polar lipids. The major FA in the TAG in all samples were 14∶0, 16∶0, and 18∶0 as saturated FA (saturates); 16∶1n−7, 18∶1n−9, and 18∶1n−7 as monoenoic FA (monoenes); and 20∶4n−6 (arachidonic acid: AA), 20∶5n−3 (EPA), and 22∶6n−3 (DHA) as PUFA. The major components found in the polar lipids were 16∶0 and 18∶0 as saturates; 22∶2n−9, 15 and 22∶2n−7, 15 as non-methylene-interrupted dienes (NMID), and AA, 22∶3n−6, 9, 15, EPA, and DHA as PUFA. Although it is a marine animal, characteristically high levels of AA were found in both the TAG and phospholipids. This result suggests that lipids of P. fucata may be influenced by those of its phytoplanktonic prey. The increase in levels of NMID from TAG to PE with a decrease in those of monoenes suggests that the tissues of this species are able to biosynthesize only the less unsaturated PUFA, such as NMID. In particular, NMID derivatives are considered to be biosynthesized in the PE; thus, they might play a particular role in the membrane, because NMID were characteristically localized only in the PE.     

Linoleic acid-induced fatty acid changes in platelet and aorta of the rat: Effect of age and cholesterol

The influence of age and cholesterol on polyunsaturated fatty acids (PUFa) levels was studied in young and old male Sprague-Dawley rats. Animals were fed a fat-free diet supplemented with 10% (by wt) safflower oil with or without 1% cholesterol for 8 wk. As a result of cholesterol feeding, proportions of linoleic acid (18∶2n−6) and dihomo-γ-linolenic acid (30∶3n−6) were increased and and that of arachidonic acid (20∶4n−6) was decreased in the liver and platelet phospholipids in 64-wk-old rats, suggesting inhibitory effects of cholesterol on 20∶4n−6 synthesis from 18∶2n−6. The prominent age-dependent effect on the levels of PUFA was a retention of C−22 n−3 PUFA, accompanied by decreased C−22 n−6 PUFA and increased 20∶3n−6 in the liver and platelet phospholipids. Ratio of 20∶3n−6/20∶4n−6 increased in 64-wk-old rats regardless of dietary cholesterol, suggesting depressed Δ5-desaturase with age. In aorta phospholipids, 20∶3n−6 content and 20∶3n−6/20∶4n−6 ratio increased with cholesterol supplementation, but not with age. These results suggest that changes of PUFA composition of platelet phospholipids with age are closely linked with changes in liver phospholipids. The 20∶4n−6 content in both platelet and aorta phospholipids is kept constant, despite other n−6 and n−3 PUFA being affected by age.     

Supplementation and delivery of n−3 fatty acids through spray-dried milk reduce serum and liver lipids in rats

Indian diets comprising staples such as cereals, millets, and pulses provide 4.8 energy % from linoleic acid (18∶2n−6) but fail to deliver adequate amounts of n−3 FA. Consumption of long-chain n−3 PUFA such as EPA (20∶5n−3) and DHA (22∶6n−3) is restricted to those who consume fish. The majority of the Indian population, however, are vegetarians needing additional dietary sources of n−3 PUFA. The present work was designed to use n−3 FA-enriched spray-dired milk powder to provide n−3 FA. Whole milk was supplemented with linseed oil to provide α-linolenic acid (LNA, 18∶3n−3), with fish oil to provide EPA and DHA, or with groundnut oil (GNO), which is devoid of n−3 PUFA, and then spray-dired. Male Wistar rats were fed the spray-dired milk formulations for 60 d. The rats given formulations containing n−3 FA showed significant increases (P<0.001) in the levels of LNA or EPA/DHA in the serum and in tissue as compared with those fed the GNO control formulation. Rats fed formulations containing n−3 FA had 30–35% lower levels of serum total cholesterol and 25–30% lower levels of serum TAG than control animals. Total cholesterol and TAG in the livers of rats fed the formulations containing n−3 FA were lower by 18–30% and 11–18%, respectively, compared with control animals. This study showed that spray-dried milk formulations supplemented with n−3 FA are an effective means of improving dietary n−3 FA intake, which may decrease the risk factors associated with cardiovascular disease.