The influence of fish, meat and polyunsaturated fat intakes on platelet phopholipid polyunsaturated fatty acid in male Melbourne Chinese and Caucasian

Objective: The aims of this study were to investigate (1) platelet phospholipid (PL) polyunsaturated fatty acid (PUFA) composition in subjects who were the Melbourne Chinese migrants, compared with those who were the Melbourne Caucasians and (2) the relationship between platelet PL PUFA and intake of fish, meat and PUFA.

Design: Cross-sectional comparison of the Melbourne Chinese and Caucasians.

Setting: Free-living male subjects.

Subjects: Ninety-seven Melbourne Chinese migrants and 78 Melbourne Caucasians who were recruited in Melbourne.

Outcome measures: Dietary intake was assessed using a semi-quantitative food frequency questionnaire. The platelet PUFA was measured by gas-liquid chromatography.

Results: The Melbourne Chinese had significantly higher proportions of platelet PL 20:5n-3 (P=0.006), 22:6n-3 (P<0.0001), total n-3 (P=0.027) and 22:5n-6 (P=0.0002), and a significantly higher intake of fish (P=0.012) and white meat (P=0.0045) compared with the Melbourne Caucasians. In addition, the Melbourne Chinese had significantly lower proportions of 20:3n-6 (P=0.023), 20:4n-6 (P<0.002), 22:4n-6 (P<0.0001), total n-6 (P=0.037), 22:5n-3 (P<0.0001) and ratio of n-6/n-3 (P=0.011), and a significantly lower intake of red and total meat (P<0.0001) than the Melbourne Caucasians. Fish consumption was significantly positively correlated with platelet PL 20:5n-3 and 22:6n-3, and significantly negatively correlated with 22:5n-3 (P<0.05). Meat consumption was significantly positively correlated with 22:5n-3 and significantly negatively correlated with 22:5n-6, 20:5n-3 and 22:6n-3 (P<0.05). Dietary PUFA intake was significantly positively correlated with 20:3n-6, 22:4n-6 and 22:5n-3, and significantly negatively correlated with 22:5n-6, 20:5n-3 and 22:6n-3 (P<0.05).

Conclusions: Compared with Caucasians, the Melbourne Chinese had a significantly higher level of platelet PL n-3 PUFA, which might contribute to the low CVD mortality in this population. Platelet PL 20:5n-3 and 22:6n-3 were significantly positively correlated with fish intake, and negatively significantly correlated with dietary intake of meat and PUFA, while 22:5n-3 was significantly positively correlated with dietary meat and PUFA intake, and significantly negatively correlated with fish intake. Dietary intake of PUFA and fish are potential confounding factors for assessing the effects of meat consumption on platelet PL individual PUFA. Dietary intake of PUFA and meat did not influence the incorporation of fish long chain n-3 PUFA to platelet PL in this study population.

Effect of feeding systems on omega-3 fatty acids, conjugated linoleic acid and trans fatty acids in Australian beef cuts: potential impact on humnan health

The influence of feeding systems on the levels of functional lipids and other fatty acid concentrations in Australian beef was examined. Rump, strip loin and blade cuts obtained from grass feeding, short-term grain feeding (80 days; STGF) and long-term grain feedlot rations (150-200 days; LTFL) were used in the present study. The typical Australian feedlot ration contains more than 50% barley and/or sorghum and balanced with whole cottonseed and protein meals were used as feed for STGF and LTFL regimens. Meat cuts from 18 cattle for each feeding regimen were trimmed of visible fat and  connective tissue and then minced (300 g lean beef); replicate samples of 7g were used for fatty acid (FA) analysis. There was a significantly higher level of total omega-3 (n-3) and long chain n-3 FA in grass-fed beef (P <0.0001) than the grain-fed groups regardless of cut types. Cuts from STGF beef had significantly reduced levels of n-3 FA and conjugated linoleic acid (CLA) and similar levels of saturated, monounsaturated and n-6 FA compared with grass feeding (P <0.001). Cuts from LTFL beef had higher levels of saturated, monounsaturated, n-6 FA and trans 18:1 than similar  cuts from the other two groups (P <0.01), indicating that increased length of grain feeding was associated with more fat deposited in the carcass. There was a step-wise increase in trans 18:1 content from grass to STGF to LTGF, suggesting grain feeding elevates trans FA in beef, probably because of increased intake of 18:2n-6. Only grass-fed beef reached the target of more than 30mg of long chain n-3 FA/100 g muscle as recommended by Food Standard Australia and New Zealand for a food to be considered a source of omega- 3 fatty acids. The proportions of trans 18:1 and n-6 FA were higher (P<0.001) for both grain-fed beef groups than grass-fed beef. Data from the present study show that grain feeding decreases functional lipid  components (long chain n-3 FA and CLA) in Australian beef regardless of meat cuts, while increasing total trans 18:1 and saturated FA levels.

No effect of n - 3 long-chain polyunsaturated fatty acid (EPA and DHA) supplementation on depressed mood and cognitive function : a randomised controlled trial

Low dietary intakes of the n-3 long-chain PUFA (LCPUFA) EPA and DHA are thought to be associated with increased risk for a variety of adverse  outcomes, including some psychiatric disorders. Evidence from  observational and intervention studies for a role of n-3 LCPUFA in depression is mixed, with some support for a benefit of EPA and/or DHA in major depressive illness. The present study was a double-blind randomised controlled trial that evaluated the effects of EPA+DHA supplementation (1.5 g/d) on mood and cognitive function in mild to moderately depressed  individuals. Of 218 participants who entered the trial, 190 completed the planned 12 weeks intervention. Compliance, confirmed by plasma fatty acid concentrations, was good, but there was no evidence of a difference between supplemented and placebo groups in the primary outcome - namely, the depression subscale of the Depression Anxiety and Stress Scales at 12 weeks. Mean depression score was 8.4 for the EPA+DHA group and 9.6 for the placebo group, with an adjusted difference of - 1.0 (95 % CI - 2.8, 0.8; P = 0.27). Other measures of mood, mental health and cognitive function, including Beck Depression Inventory score and attentional bias toward threat words, were similarly little affected by the intervention. In conclusion, substantially increasing EPA+DHA intake for 3 months was found not to have beneficial or harmful effects on mood in mild to moderate depression. Adding the present result to a meta-analysis of previous relevant randomised controlled trial results confirmed an overall negligible benefit of n-3 LCPUFA supplementation for depressed mood.

Bovine milk fat enriched in conjugated linoleic and vaccenic acids attenuates allergic airway disease in mice

Background: It has been argued that a reduction in the Western diet of anti-inflammatory unsaturated lipids, such as n-3 polyunsaturated fatty acids, has contributed to the increase in the frequency and severity of allergic diseases.

Objective: We investigated whether feeding milk fat enriched in conjugated linoleic acid and vaccenic acids (VAs) ('enriched' milk fat), produced by supplementing the diet of pasture-fed cows with fish and sunflower oil, will prevent development of allergic airway responses.

Methods: C57BL/6 mice were fed a control diet containing soybean oil and diets supplemented with milk lipids. They were sensitized by intraperitoneal injection of ovalbumin (OVA) on days 14 and 28, and challenged intranasally with OVA on day 42. Bronchoalveolar lavage fluid, lung tissues and serum samples were collected 6 days after the intranasal challenge.

Results: Feeding of enriched milk fat led to marked suppression of airway inflammation as evidenced by reductions in eosinophilia and lymphocytosis in the airways, compared with feeding of normal milk fat and control diet. Enriched milk fat significantly reduced circulating allergen-specific IgE and IgG1 levels, together with reductions in bronchoalveolar lavage fluid of IL-5 and CCL11. Treatment significantly inhibited changes in the airway including airway epithelial cell hypertrophy, goblet cell metaplasia and mucus hypersecretion. The two major components of enriched milk fat, cis-9, trans-11 conjugated linoleic acid and VA, inhibited airway inflammation when fed together to mice, whereas alone they were not effective.

Conclusion: Milk fat enriched in conjugated linoleic and VAs suppresses inflammation and changes to the airways in an animal model of allergic airway disease.

The role of fatty acids in satiation and satiety

Background – Satiation and satiety describe the events which lead to meal termination and the maintenance of hunger induced by physical and metabolic events following food ingestion. Fatty acids, components of dietary fat (triglyceride) may be important, if not essential components of satiation and satiety. Emerging evidence suggests fatty acid now constitutes a sixth taste modality and orally sensed fatty acids mediate unique cephalic and hormonal responses priming the body for fat digestion, and may contribute to sensory specific satiety. Once ingested, fatty acids are sensed in the gastrointestinal tract (GIT) where they cause the release of hormones, stimulate the vagus and enter the blood stream where they act a number of organs (brain, liver) to influence satiety.
Objective – To review the role of fatty acids in sensory and metabolic satiation and satiety.
Design – Literature search and review of papers from the past decade on satiety, satiation, fat taste and fatty acids.
Outcomes – The physiological significance of gustatory fat detection is still unclear, but it may signal the nutritious content of fat similar to the tastes of sweet or umami which signal the presence of carbohydrate or proteins. Like other tastants, fatty acid taste sensitivity is thought to vary in the population and differences in sensitivity may influence dietary choice and fat intake. Fatty acid taste may contribute to sensory specific satiety as foods are eaten. Animal models have observed an inverse relationship between oral fatty acid sensitivity and fat consumption, which leads to obesity. Observations that the obese have heightened preferences for, and consume more fat than lean individuals questions whether such a relationship may also be apparent in humans. At the GIT, fatty acids are sensed by enterocytes and bind to receptors, transporters or ion channels where they initiate gut-brain communication over nutrient status through the vagus and cause the release of satiety hormones which lead to meal termination. Inefficient fatty acid sensing at either or both locations is thought to accompany the aetiology of obesity.
Conclusion – Variations in sensitivity to fatty acids may alter preferences and consumption of fats or hormonal responses to fat ingestion which influence sensory-specific, metabolic and subjective satiety.

Isolation and characterization of polyunsaturated fatty acid producing Thraustochytrium species : screening of strains and optimization of omega-3 production

An isolation program targeting Thraustochytrids (marine fungoid protists) from 19 different Atlantic Canadian locations was performed. Sixty-eight isolates were screened for biomass, total fatty acid (TFA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) content. Analysis of fatty acid methyl ester results discerned four distinctive clusters based on fatty acid profiles, with biomass ranging from 0.1 to 2.3 g L−1, and lipid, EPA, and DHA contents ranging from 27.1 to 321.14, 2.97 to 21.25, and 5.18 to 83.63 mg g−1 biomass, respectively. ONC-T18, was subsequently chosen for further manipulations. Identified using 18S rRNA gene sequencing techniques as a Thraustochytrium sp., most closely related to Thraustochytrium striatum T91-6, ONC-T18 produced up to 28.0 g L−1 biomass, 81.7% TFA, 31.4% (w/w biomass) DHA, and 4.6 g L−1 DHA under optimal fermentation conditions. Furthermore, this strain was found to produce the carotenoids and xanthophylls astaxanthin, zeaxanthin, canthaxanthin, echinenone, and β-carotene. Given this strain’s impressive productivity when compared to commercial strains, such as Schizochytrium sp. SR21 (which has only 50% TFA), coupled with its ability to grow at economical nitrogen and very low salt concentrations (2 g L−1), ONC-T18 is seen as an ideal candidate for both scale-up and commercialization.

Evaluation of fatty acid extraction methods for Thraustochytrium sp. ONC-T18

Various extraction methods were assessed in their capacity to extract fatty acids from a dried biomass of Thraustochytrium sp. ONC-T18. Direct saponification using KOH in ethanol or in hexane:ethanol was one of the most efficient techniques to extract lipids (697 mg g^-1). The highest amount of fatty acids (714 mg g^-1) was extracted using a miniaturized Bligh and Dyer extraction technique. The use of ultrasonics to break down cell walls while extracting with solvents (methanol:chloroform) also offered high extraction yields of fatty acids (609 mg g^-1). Moreover, when the transesterification mixture used for a direct transesterification method was doubled, the extraction of fatty acids increased approximately 77% (from 392 to 696 mg g^-1). This work showed that Thraustochytrium sp. ONC-T18 has the ability to produce over 700 mg g^-1 of lipids, including more than 165 mg g-1 of docosahexaenoic acid, which makes this microorganism a potential candidate for the commercial production of polyunsaturated fatty acids. Finally, other lipids, such as myristic, palmitic, palmitoleic, and oleic acids, were also produced and recovered in significant amounts (54, 196, 123, and 81 mg g^-1), respectively.

Fatty acid composition in tissues of mice fed diets containing conjugated linolenic acid and conjugated linoleic acid

The influence of 1% alpha-eleostearic acid (α-ESA, cis9,trans11,trans 13-18:3) and 1% punicic acid (PA, cis9,trans11,cis13-18:3) on fatty acid composition in mouse tissues was compared with conjugated linoleic acid (CLA, mixture of primarily cis9,trans11- and trans10,cis12-18:2) in the present study. The content (% total fatty acids) of 18:2n-6 was significantly reduced in the heart and adipose tissues, and total polyunsaturated fatty acids (PUFAs) and n-6 PUFA were significantly reduced in adipose tissue by α-ESA, PA and CLA feeding. The content of 22:6n-3 and total n-3 PUFA were significantly increased in the liver, kidney and heart by PA feeding, but not by α-ESA. In contrast to PA, supplementation with CLA significantly decreased 22:6n-3 in the liver, kidney and heart. The content of 20:4n-6 was significantly decreased in the liver and kidney by CLA feeding, but not by α-ESA and PA. The present results indicate that α-ESA, PA and CLA have differential effects on 22:6n-3 and 20:4n-6 content in mouse tissues.

Polyunsaturated fats in meat from Merino, first- and second-cross sheep slaughtered as yearlings

This study examined the level of long chain omega-3 and omega-6 polyunsaturated fats, the ratio of polyunsaturated fat to saturated fat (PUFA/SFA) and the ratio of omega-6 to omega-3 (n-6/n-3) fat in sheep grown under grazing conditions in Australia. The sheep genotypes used were Poll Dorset_growth × Border Leicester Merino (PDg × BLM), Poll Dorset_growth × Merino (PDg × M), Poll Dorsetmuscling × Merino (PDm × M), Border Leicester × Merino (BL × M) and Merino × Merino (M × M). Loin muscles (Longissimus lumborum) collected from 40 ewe and wether sheep slaughtered at 14 months of age were processed for fatty acid determination. After frozen storage, 20 g samples were minced and a 7 g homogenate was processed for muscle lipid extraction using a chloroform:methanol (2:1) procedure. There was an increase in PUFA/SFA as the proportion of Merino genetics increased in the progeny (second-cross < first-cross < Merino), but this was not shown in the n-6/n-3 ratio. The PUFA/SFA trend appeared to be associated with an increase in the level of total polyunsaturated fats, but not a decrease in the level of total saturated fats. The results demonstrate that there is a need to improve the PUFA/SFA content in first- and second-cross animals which are mainly used for meat production in Australia so as to maintain the healthy lipids in meat. Nutritional manipulation through feeding systems or selection of sires for greater heritability of omega-3 fat deposition may be suitable pathways to elevate the ratio of polyunsaturated fatty acids, and in particular omega-3.

Transesterification of fish oil to produce fatty acid ethyl esters using ultrasonic energy

This study evaluated the production of fatty acid ethyl esters from fish oil using ultrasonic energy and alkaline catalysts dissolved in ethanol. The feasibility of fatty acid ethyl ester production was determined using an ultrasonic bath and probe, and between 0.5 and 1% KOH (added to the fish oil). Furthermore, factors such as ultrasonic device (bath and probe), catalyst (KOH and C2H5ONa), temperature (20 and 60 °C), and duration of exposure (10–90 min) were assessed. Sodium ethoxide was found to be a more efficient catalyst than KOH when transesterifying fish oil. Ultrasonic energy applied for greater than 30 min at 60 °C using 0.8% of C2H5ONa as a catalyst transesterified over 98% fish oil triglycerides to fatty acid ethyl esters. It is reasonable to conclude that the yield of fatty acid ethyl esters produced by applying ultrasonic energy to fish oil is related to the sonication time. Due to increases in the surface area contact between the reactants and the catalyst, ultrasonic energy has the potential to reduce the production time required by a conventional large-scale commercial transesterification method that uses agitation as a way of mixing.

Optimization of fatty acid determination in selected fish and microalgal oils

The use of ten fatty acid methyl ester reference standards coupled with a detailed quantification method was shown to significantly optimize the fatty acid determination of selected fish and microalgal oils when compared to methods that use only one reference standard (C19:0 or C23:0) as a relative response factor. When using the mixture of ten reference standards after transesterifying oils with NaOH/BF3, determination of total fatty acids, eicosapentaenoic acid and docosahexaenoic acid improved by an average of 7.3, 11.5 and 8.4%, respectively. Furthermore, improvements of 13.9, 18.9 and 6.8% of total fatty acids, EPA and DHA, respectively, were obtained when using the mixture of reference standards for fatty acid determination after directly extracting and transesterifying oil contained in microalgal cells with a mixture of methanol, HCl and chloroform. Fatty acid methyl ester standards dissolved in isooctane showed <5% variability throughout 130 days of stability testing when stored at −20 °C. The optimized method can be used for improving the quantification of fatty acids in both oils (fish and microalgal oils) and dry microalgal cells.

Production and bioavailability of calcium and magnesium salts of omega-3 fatty acids

In order to provide an alternative to traditional liquid fish oil gelatin capsules, we developed a solid, powdered form of omega-3 fish oil concentrate by forming calcium- and magnesium-fatty acid salts. These salts were produced using a concentrated fish oil ethyl ester that contained in excess of 60% omega-3 fatty acids. The bioavailability of these omega-3 salts was compared with that of fish oil ethyl ester in mice. Animals were given 8 mg of omega-3 fatty acid ethyl ester concentrate (control), calcium- or magnesium-omega-3 salts daily for three weeks. The omega-3 salt products resulted in omega-3 fatty acid content in serum and red blood cell membranes comparable to that produced by the ethyl ester supplementation. In addition, fecal excretion of omega-3 fatty acids was not increased by the presence of calcium or magnesium. In fact, there was a tendency for less omega-3 fatty acids to be excreted.