Polyunsaturated fatty acids (PUFAs) content of the fungus Mortierella alpina isolated from soil

Twenty-five isolates of Mortierella spcies were prepared, which can be used for the production of polyunsaturated fatty acids (PUFAs)-rich oil for nutritional supplements. The fatty acid contents were determined after heterotrophic fermentation. The content of total fatty acids (TFAs) in the cell dry weight of all isolates including two commercially purchased Mortierella alpina strains ranged from 207.51 to 370.11 mg/g, whereas PUFAs were the dominant fatty acid type. The highest PUFA-containing strain, M. alpina SC9, was identified and confirmed as a new strain of M. alpina through comparison analysis of the sequences of internal transcribed spacers 1 and 2 (ITS1 and ITS2) and the 5.8S rDNA region. During a 7-day fermentation, the PUFAs content of M. alpina SC9 varied between 189.83 and 240.00 mg/g, with a remarkable correlation between the oleic acid (C18:1, OA) and arachidonic acid (C20:4n-6, AA) contents and between the linoleic acid (C18:2n-6, LA) and AA contents, suggesting the PUFA content in the fungus is tightly regulated. This study provides a framework of isolation, identification, and characterization of an important PUFA-producing species, M. alpina.     

Effect of unialgal diets on the composition of fatty acids and sterols in juvenile ark shell Tegillarca granosa Linnaeus

This study has investigated the effects of six different unialgal diets ( Chaetoceros calcitrans , Platymonas helgolandica , Chlorella sp., Isochrysis galbana , Nannochloropsis oculata , and Pavlova viridis ) on the composition of fatty acids and sterols in juvenile ark shell Tegillarca granosa Linnaeus. The best feeding effects on the growth of shellfish were found in C. calcitrans, followed by I. galbana and P. viridis, whereas Chlorella sp. and N. oculata exhibited relatively poor effects. The fatty acid and sterol compositions in the six microalgae and the juvenile ark shell after feeding were analyzed, and 39 fatty acids and 18 sterols were identified. Although the results demonstrate a close correlation between the sterol compositions in algal species and juvenile ark shell, a similar correlation was not observed between fatty acids. In the juvenile ark shell fed microalgae, the ratio of total saturated fatty acids (SFA) rapidly decreases, whereas the proportion of total polyunsaturated fatty acids (PUFAs) increases considerably. The abundances of AA, EPA, and DHA increase most significantly in shellfish with better growth (fed C. calcitrans, I. galbana, and P. viridis). The number of sterol species is reduced, but the total sterol content in groups fed corresponding microalgae increases, and abundant plant sterols, instead of cholesterol, are accumulated in juvenile ark shell fed appropriate microalgae I. galbana and P. viridis. Therefore, to be more conducive to human health, I. galbana and P. viridis, of the six experimental microalgae, are recommended for artificial ark shell culture.     

Seasonal changes in crude and lipid composition of herring fillets,byproducts, and respective produced oils

Crude and fatty acid composition analyses were performed on fillets, byproducts, and oil originating from herring (Clupea harengus) caught off the North Sea from June 1999 to January 2001. Monthly statistical differences were found in the fat content, the range of variation being larger in fillets than in byproducts. The most consistent change observed in fillets was an increase of unsaturation from May to September reflected in a reduced percentage of monounsaturated fatty acids, whereas for byproducts and oil this trend was not so well defined. The results indicated that the lowest values of the total amount of polyunsaturated fatty acids (PUFAs) in the oil were found from January to March (approximately 14%), coinciding with the postspawning and starvation period. In contrast, the highest values were found from June to August (approximately 23%). Thus, the herring byproducts are all year an adequate raw material for fish oil production; however, during the summer they are richer in PUFAs.     

Influence of drying and storage on lipid and carotenoid stability of the microalga Phaeodactylum tricornutum

The influence of short-term storage and spray- and freeze-drying of fresh microalgal paste on the stability of lipids and carotenoids of Phaeodactylum tricornutum was investigated. Furthermore, the effects of storage time (14 and 35 days) and condition (vacuum packed vs non vacuum packed, -20 °C vs 4 °C vs 20 °C) after spray- and freeze-drying were studied. Total lipid content, free fatty acid content, carotenoid content and degree of lipid oxidation were measured. No effects of spray- and freeze-drying and subsequent storage were found on total lipid content, except for short-term storage of the fresh microalgal paste, which led to pronounced lipolysis and therefore a lower total lipid content. Freeze-dried microalgae were found to be more susceptible to lipolysis upon storage than spray-dried microalgae. On the other hand, spray-dried microalgae were more susceptible to oxidation than freeze-dried microalgae, possibly due to breakdown of protecting carotenoids upon spray-drying. Hardly any effect of storage condition was observed for any of the parameters tested.     

Phytochemical changes induced by different nitrogen supply forms and radiation levels in two leafy Brassica species

The effect of three different nitrogen (N) supply forms differing in their ammonium-to-nitrate (NH(4):NO(3)) ratio (100% NH(4), 50% NH(4) + 50% NO(3), 100% NO(3)) under three different levels of daily photosynthetic active radiation (PAR) (low, 5.0; medium, 6.8; high, 9.0 mol m(-2) day(-1)) on a range of desirable health-promoting phytochemicals in Brassica rapa subsp. nipposinica var. chinoleifera and Brassica juncea was determined. The 100% NH(4) supply under medium PAR levels led to the highest concentration of glucosinolates based on a low nitrogen/sulfur ratio as well as high levels of carotenoids in the leaves of both Brassica species. However, the 100% NH(4) supply under low and medium PAR levels resulted in low concentrations of flavonoids based on high N concentration in the leaves. Thus, the data provided here have strong implications for crop management strategies aimed at optimizing both the concentration and composition of a range of phytochemicals.     

Oxidative degradation of lipids during mashing

Although hardly any polyunsaturated fatty acids (PUFAs) are present in the endproduct, the ingredients used for the production of beer contain a high concentration of PUFAs, such as linolic and linolenic acid. These compounds are readily oxidized, resulting in the formation of lipid-derived products that reduce the taste and quality of beer enormously. During mashing relatively high amounts of PUFAs are exposed to atmospheric oxygen at a relatively high temperature. This makes mashing a critical step in the brewing process with regard to the formation of lipid-derived off-taste products. F1 phytoprostane (PPF1) changes in antioxidant capacity and monohydroxy fatty acids (OH-FAs) were used as markers for the detection of oxidative damage to fatty acids during mashing. The pattern of OH-FA formation indicates that enzymatic oxidation of PUFAs is more important than nonenzymatic oxidation during the mashing process. Nevertheless, substantial nonenzymatic radical formation is evident from the increase of specific OH-FAs and PPF1s. It was found that a low oxygen tension reduces oxidative damage and gives a high antioxidant capacity of the mashing mixture. This indicates that mashing should be done under low oxygen pressure.     

Comparison of two microalgal diets. 1. Influence on the biochemical and fatty acid compositions of raw oysters (Crassostrea gigas)

Oyster farming in France is a traditional activity. Each year, 149 000 tons of oysters are fattened before being sold. More and more oyster farmers supplement the diet of oysters by microalgae to optimize the fattening process and to improve both the growth and flesh quality of oysters. In the present study, oysters were supplemented by two microalgae: Skeletonema costatum and Tahitian isochrysis clone. The ash, protein, carbohydrate (including glycogen), and lipid contents were analyzed. The growth of oysters was improved when they were fattened with microalgae, leading to an increase in the condition index. The chemical composition of oysters was influenced by the chemical composition of the microalgae, especially the level of glycogen, which was significantly increased for oysters supplemented by microalgae. The fatty acid profile of oysters fattened by microalgae was positively correlated with the fatty acid profiles of S. costatum and T. isochrysis. These results show the effectiveness of supplementation by microalgae on the growth and on the biochemical composition (glycogen and fatty acids especially) of oysters.     

Individual- and scattered-tree influences on ultraviolet irradiance

Many of the potential effects of ultraviolet radiation (UVR)—damage to materials, altered herbivory of insects and activity of microbes, modified growth of vegetation, and adverse or beneficial effects on human health—are modified by the presence of trees that influence UVR exposure to various degrees. Though tree effects on total solar irradiance have been investigated often by measurements and modeling, tree influences on UVR, particularly in the ultraviolet B (UVB, 320–280 nm), differ substantially from tree influences on the rest of the solar spectrum, and thus the ratio of UVB to photosynthetically active radiation (PAR) is altered. Trees greatly reduce both UVB and PAR irradiance in their shade when they obscure both the sun and sky. Beneath dense forest canopies, relative irradiance (I_r=irradiancebeneath trees/above-canopy irradiance) for both UVB and PAR radiation may be 0.01–0.02. In the shade of a single tree, I_r on the horizontal in the PAR and total shortwave (SW) was about 0.1, whereas in the UVB and ultraviolet A (UVA, 320–400 nm), I_r was about 0.4. Conversely, where direct beam radiation came through gaps between crowns in a group of deciduous trees in winter, PAR I_r values averaged 0.95 and UVB I_r averaged only 0.41. In comparisons of minimum values of I_r on horizontal and the south-facing vertical surfaces in tree shade for UVB, UVA, SW, and PAR, where UVB I_r on the horizontal ranged from 0.22 to 0.62, depending on solar zenith angle, UVB I_r on the vertical ranged from 0.05 to 0.27. UVB I_r consistently exceeded UVA I_r on both the horizontal and vertical surfaces. PAR and SW I_r differed little between horizontal and vertical surfaces in tree shade. Modeled average I_r on the horizontal below a regular grid of ellipsoidal tree crowns was given byI_p=1−m−(θ^0.711/5.05)sin(πm), where m is fraction of area covered by tree crowns and θ is solar zenith angle. The tree influences will vary with pollutants in the boundary layer, which affect scattering of UVR.     

Restoration of fillet n-3 long-chain polyunsaturated fatty acid is improved by a modified fish oil finishing diet strategy for atlantic salmon (Salmo salar L.) smolts fed palm fatty acid distillate

Reducing the lipid content in fish prior to feeding a fish oil finishing diet (FOFD) has the potential to improve n-3 long-chain (≥ C(20)) polyunsaturated fatty acid (LC-PUFA) restoration. This study had two main objectives: (1) determine whether feeding Atlantic salmon smolt a 75% palm fatty acid distillate diet (75PFAD) improves the apparent digestibility (AD) of saturated fatty acids (SFA) and (2) examine whether a food deprivation period after growth on 75PFAD leads to higher n-3 LC-PUFA restoration in the fillet when applying a FOFD. The AD of SFA was higher for 75PFAD compared to that of a fish oil (FO) diet. The relative level (as % total fatty acids (FA)) of n-3 LC-PUFA was higher in unfed fish compared to that in continuously fed fish after 21 and 28 day FOFD periods, respectively. Our results suggest that a food deprivation period prior to feeding a FOFD improves the efficiency of n-3 LC-PUFA restoration in the fillet of Atlantic salmon smolt.     

Relevance of isotopic and molecular biomarkers for the authentication of milk according to production zone and type of feeding of the cow

The first objective of the present paper was to assess the potential of both isotopic ( (18)O/ (16)O in milk water) and molecular biomarkers (terpenes, fatty acids, carotenoids, and vitamins) and milk color to discriminate the production zone (lowland or upland areas) from which 49 tanker bulk milks were collected over one year from a total of 204 farms. The milk water (18)O enrichment was higher in lowland (<500 m altitude) than in upland (>700 m altitude), but the delta (18)O values failed to discriminate systematically the production zone at the scale of the year because of its high variability related to the sampling period. In contrast with vitamins A and E, carotenoids, and milk color measurements, terpenes and fatty acids were confirmed to be relevant tracers of the production zone. The milk compounds with the strongest discriminative potential were fatty acids, which were determined by high-resolution gas chromatography. The calculation of fatty acid ratios, which permits the limitation of using fatty acid relative quantity expressed in percentage of total fatty acids to be overcome, was shown to be particularly relevant in discriminating upland from lowland milk ratios. The selection of two pairs of ratios, namely, iso-C17:0/C18:3 n-3 and iso-C15:0/iso-C14:0, enabled the authentication of 100% of the highland versus lowland milks whatever the season. The second objective was to evaluate the relevance of fatty acid composition to discriminate milks according to the proportion of corn silage in the diets of dairy cows. The selection of two fatty acids ratios, namely, trans11 cis15-C18:2/trans11-C18:1 and cis9-C16:1/iso-C16:0, enabled the correct classification of 100% of the milk samples according to the proportion of corn silage in the basic fodder rations (<25% vs >30%). The relationship between the milk production zone and the type of forage fed to the cows is discussed.     

Determination of carotenoids in spear shrimp shells (Parapenaeopsis hardwickii) by liquid chromatography

The objectives of this study were to develop a high-performance liquid chromatography method for analysis of carotenoids in spear shrimp shells (Parapenaeopsis hardwickii) and to compare the extraction efficiency of carotenoids by supercritical carbon dioxide (SCD) and solvents. Results showed that the most appropriate HPLC method was accomplished by employing a Cosmosil 5C18-AR-II column and a mobile phase of methanol-dichloromethane-acetonitrile (90:5:5, v/v/v) (A) and water (100%) (B) with the following gradient elution: 92% A and 8% B in the beginning, decreased to 4% B in 9.5 min, 1% B in 26 min, 0% B in 35 min, maintained for 25 min, and returned to 92% A and 8% B in 61 min. All-trans-astaxanthin and its two cis isomers, as well as five astaxanthin monoesters and 11 diesters were resolved within 60 min with a flow rate at 2 mL/min and detection at 480 nm. Astaxanthin diesters were found to contain 12 fatty acids, of which palmitic acid and stearic acid constituted a large portion, whereas astaxanthin monoesters were found to contain 10 fatty acids with arachidonic acid dominating. Solvent extraction could generate a higher content of trans-astaxanthin and astaxanthin esters, while SCD extraction could produce greater levels of 9-cis-astaxanthin and 13-cis-astaxanthin.     

Fiber reduction and lipid enrichment in carotenoid-enriched distillers dried grain with solubles produced by secondary fermentation of Phaffia rhodozyma and Sporobolomyces roseus

Carotenoid-enriched distillers dried grain with solubles (DDGS) developed as a value-added animal feed to provide carotenoids from mono and mixed culture (Mx) fermentation of red yeasts Phaffia rhodozyma (PR) and Sporobolomyces roseus (SR) were evaluated for their nutritional composition and compared to the control (C) DDGS. Apart from providing carotenoids, all three fermentation treatments reduced fiber with best reduction of 77% in PR, enhanced crude fat with highest of 81% in Mx, and reduced protein, amino acids and nitrogen by 50% in PR. DDGS fiber reduction by 77% was achieved by P. rhodozyma in the absence of any pretreatment. Qualitative and quantitative differences in fatty acid profiles were seen among the treatments. Vaccenic acid, a monounsaturated fatty acid produced in SR and Mx fermentation, was absent in C and PR. All these nutritional modifications are highly desirable in different DDGS-based animal feeds and can be explored to obtain tailor-made feeds/feed blends for specific animal diets.     

植物中γ-亚麻酸合成关键酶——△^6-脂肪酸脱氢酶

γ-亚麻酸作为人体必需的不饱和脂肪酸，对人体的激素调节及脂肪酸代谢发挥着重要的生理作用。△^6-脂肪酸脱氢酶是多不饱和脂肪酸合成途径中的限速酶。本文介绍了不饱和脂肪酸γ-亚麻酸代谢途径中的关键酶△^6-脂肪酸脱氢酶的结构功能与目前△^6-脂肪酸脱氢酶的基因工程研究进展，并对其应用进行了展望，以期为利用基因工程手段生产γ-亚麻酸提供参考。     

Effect of the polyunsaturated fatty acid composition of beef muscle on the profile of aroma volatiles

The effect of n-3 polyunsaturated fatty acids (PUFAs) in beef muscle on the composition of the aroma volatiles produced during cooking was measured. The meat was obtained from groups of steers fed different supplementary fats: (i) a palm-oil-based control; (ii) bruised whole linseed, which increased muscle levels of alpha-linolenic (C18:3 n-3) and eicosapentaenoic acid (EPA, C20:5 n-3); (iii) fish oil, which increased EPA and docosahexaenoic acid (C22:6 n-3); (iv) equal quantities of linseed and fish oil. Higher levels of lipid oxidation products were found in the aroma extracts of all of the steaks with increased PUFA content, after cooking. In particular, n-alkanals, 2-alkenals, 1-alkanols, and alkylfurans were increased up to 4-fold. Most of these compounds were derived from the autoxidation of the more abundant mono- and di-unsaturated fatty acids during cooking, and such autoxidation appeared to be promoted by increased levels of PUFAs.     

Isolation of two anti-inflammatory and one pro-inflammatory polyunsaturated fatty acids from the brown seaweed Undaria pinnatifida

Two anti-inflammatory omega-3 polyunsaturated fatty acids (PUFAs) of stearidonic acid (SA) and eicosapentaenoic acid (EPA) and one pro-inflammatory omega-6 PUFA of arachidonic acid (AA) were isolated from the edible brown seaweed Undaria pinnatifida. SA was active against mouse ear inflammation induced by phorbol myristate acetate, with IC50 values of 160, 314, and 235 microg per ear for edema, erythema, and blood flow, respectively. EPA was also active against edema, erythema, and blood flow, with IC50 values of 230, 462, and 236 microg per ear, respectively. Although AA at low concentrations showed anti-inflammatory activities when measured 10 h later, AA doses of more than 243 microg per ear induced inflammatory symptoms 1 h later. Mature thalli generally had larger amounts of PUFAs than young thalli. The algal blade contained more omega-3 PUFAs than were found in other parts, while the holdfast contained extremely high amounts of AA. Late-season thalli showed increased amounts of PUFAs, especially AA.     

Study of precursors responsible for off-flavor formation during storage of potato flakes

Off-flavors frequently appear during the storage of potato flakes. Volatile profile analysis performed by solid-phase microextraction-gas chromatography-mass spectrometry revealed that hexanal is the main compound that appears during the storage period. Hexanal may be a degradation product of linoleic acid formed through linoleic acid hydroperoxide cleavage. Profiles of hexanal precursors were determined from potato flakes at different storage time points. Linoleic acid-derived oxylipins are predominant in potato flakes. The free oxylipins identified, in descending order, are as follows: hexanal, hydroxy polyunsaturated fatty (PUFAs), oxo PUFAs, divinyl ether PUFAs, and hydroperoxy PUFAs. However, the main oxylipins detected were esterified: esterified hydroxy, hydroperoxy, and oxo PUFAs. Oxylipins reveal different evolutions during the storage period. Chiral high-performance liquid chromatography analysis of the precursors of hexanal and other oxylipins revealed a racemic composition that supports the nonenzymatic formation of hexanal and most of the other oxylipins identified.     

Effect of dietary omega 3 phosphatidylcholine on obesity-related disorders in obese Otsuka Long-Evans Tokushima fatty rats

Recent reports have shown that dietary phosphatidylcholine (PC) has various beneficial biological effects. Omega 3 polyunsaturated fatty acids (omega3 PUFAs) have also been reported to have lipid-lowering effects in animal models and human studies. In the present study, we investigated the effect of omega3 PUFAs containing PC (omega3-PC) on obesity-related disorders in Otsuka Long-Evans Tokushima fatty (OLETF) rats. Rats were fed semisynthetic diets that contained either 5% corn oil and 2% egg-PC or 5% corn oil and 2% omega3-PC for 4 weeks. During this 4 week feeding of the omega3-PC, the OLEFT rats showed a decrease of omental white adipose tissue weight. In addition, the omega3-PC diet significantly decreased liver weight and hepatic lipid levels in OLETF rats. These changes were attributable to the significant suppression of fatty acid synthase activity and significant enhancement in the activities of carnitine palmitoyltransferase and peroxisomal beta-oxidation. Moreover, the omega3-PC diet reduced serum glucose levels concomitant with the increase of serum adiponectin levels. These results show that compared with egg-PC, omega3-PC can prevent or alleviate obesity-related disorders through the suppression of fatty acid synthesis, enhancement of fatty acid beta-oxidation, and increase of the serum adiponectin level in OLETF rats.     

Volatile fatty acid production with alfalfa (Medicago sativa) and glucose in denitrifying soil at different temperatures

Summary A laboratory study was done to determine how, if at all, temperatures affect the production of volatile fatty acids and the rate of denitrification in soil. Glucose and alfalfa were compared as C substrates at temperatures of -2, 10, and 25°C under anaerobic conditions. At -2°C (soil not frozen), the denitrification rate was slow but was as rapid with alfalfa as glucose. This indicated that the production of volatile fatty acids by fermenters or other C substrates from alfalfa were adequate to sustain denitrifiers. No volatile fatty acids were apparently produced with glucose at -2°C whereas acetate and propionate were produced with alfalfa during the 26-day incubation period. During the 8-day incubation period at 10°C, there were also greater accumulations of acetate and propionate with alfalfa than with glucose. At 25°C, there was no major difference in the denitrification rate between glucose and alfalfa over a 4-day period. In a contrast to the other temperatures, more butyrate than propionate was produced at 25°C, especially with alfalfa. Acetate was the dominant volatile fatty acid produced and generally increased with temperature, especially after NO₃ exhaustion at 10 and 25°C. This indicated that acetate was a source of C for denitrifiers.     

Variation in fatty acid composition of Artemia salina nauplii enriched with microalgae and baker's yeast for use in larviculture

The high content of the essential fatty acids in some microalgae and baker's yeast has made them excellent diets for boosting the fatty acid content of livefood Artemia. The influences of baker's yeast (Saccharomyces cerevisiae) and three microalgae, viz., Chlorella salina, Chaetoceros calcitrans, and Nannochloropsis salina, were tested as diet components in marine livefeed brine shrimp Artemia salina nauplii to improve the polyunsaturated fatty acid (PUFA) composition. Artemia nauplii submerged in these diets for four different enrichment intervals (3, 6, 8, and 24 h) were found to incorporate essential fatty acids, and the percentage composition of different fatty acids was measured in the enriched Artemia nauplii and enrichment diets. N. salina produced higher levels of arachidonic acid (AA, 20:4n6, 9.50%), eicosapentaenoic acid (EPA, 20:5n3, 25.80%), and docosahexaenoic acid (DHA, 22:6n3, 4.18%) as compared to other diets. The total PUFA content of the enriched Artemia by N. salina increased by 56.50% with enrichment periods up to 8 h, followed by a significant reduction in the final 24 h. N. salina yielded Artemia nauplii with considerable EPA (8.05%), AA (14.15%), and DHA (1.85%) after 8 h of enrichment, which are significantly higher levels than in nauplii fed with the other three diets (p = 0.05). The DHA/EPA values in Artemia enriched for 6 h by N. salina and C. calcitrans were found to be, respectively, 88.46 and 25% higher than freshly hatched Artemia. Artemia enriched by C. salina and baker's yeast exhibited a reduction in PUFA content even at 6 h of enrichment. Significant relative decreases in DHA, EPA, and total PUFA in Artemia enriched with all of the diets were apparent, with a corresponding increase in the total saturated fatty acid content (26.95 +/- 9.75%) in the final stages (24 h) of enrichment (p = 0.05).     

Validity of protein, retinol, folic acid and n-3 fatty acid intakes estimated from the food-frequency questionnaire used in the Danish National Birth Cohort

OBJECTIVE: To validate intakes of protein, folic acid, retinol and n-3 fatty acids estimated from a food-frequency questionnaire in week 25 of pregnancy (FFQ-25). DESIGN: Validation was done against a 7-day weighed food diary (FD) and biomarkers of the nutrients in gestation week 32-38. SUBJECTS AND SETTING: The FFQ-25 to be validated was used in the Danish National Birth Cohort comprising 101 042 pregnant Danish women, of whom 88 participated in the present validation study. RESULTS: Estimated intakes of protein, retinol and folic acid did not differ significantly between the two dietary methods, but intake of n-3 fatty acids was one third larger when estimated from the FFQ-25. The intakes estimated from the two dietary methods were all significantly correlated, ranging from 0.20 for retinol intake to 0.57 for folic acid intake. Sensitivities of being correctly classified into low and high quintiles were between 0.22 and 0.77, and specificities were between 0.62 and 0.89. Urinary protein content did not correlate significantly with protein estimated from the FFQ (r = 0.17, P > 0.05), but did with intake estimated from the FD (r = 0.56, P < 0.0001). Erythrocyte folate correlated significantly with the estimated total intake from the FFQ (r = 0.55, P < 0.0001) and the FD (r = 0.52, P < 0.0001). No correlations with plasma retinol were found. Erythrocyte eicosapentaenoic acid (C20:5n-3) correlated significantly with n-3 fatty acids intake estimated from both the FFQ-25 (r = 0.37, P < 0.001) and the FD (r = 0.62, P < 0.0001). CONCLUSION: The FFQ-25 gives reasonable valid estimates of protein, retinol and folic acid intakes, but seems to overestimate intake of n-3 fatty acids.