C.M. Smuts

A randomised control trial in schoolchildren showed improvement in cognitive function after consuming a bread spread, containing fish flour from a marine source

BACKGROUND In humans, n-3 long-chain polyunsaturated fatty acids play a well-documented role in brain development and function. Docosahexaenoic acid and arachidonic acid are major structural components of the brain and a deficiency thereof may bring about changes in the behaviour domains of the brain. OBJECTIVE This trial investigated the effect of an experimental fish-flour bread spread rich in n-3 long-chain polyunsaturated fatty acids, on cognition of children (7-9yr). DESIGN Subjects (n=183) were randomly assigned to an experimental (n=91) and control group (n=92), receiving either the fish-flour spread or a placebo spread for 6 months in a single-blind study. Plasma and red blood cell phospholipid fatty acid composition and cognition were measured at baseline and post-intervention. RESULTS After the intervention, eicosapentaenoic and docosahexaenoic acid levels were significantly higher in the experimental group compared to the control group (p< 0.0001). Significant intervention effects were also observed for the Hopkins Verbal Learning Test Recognition (estimated effect size: 0.80; 95% confidence interval (CI): 0.15; 1.45) and Discrimination Index (estimated effect size: 1.10; 95% CI: 0.30; 1.91), as well as the Spelling test (estimated effect size: 2.81; 95% CI: 0.59; 5.02) by both per protocol and intention to treat analyses. A marginally significant (p=0.0646) effect was observed for the Reading test (estimated effect size: 2.21; 95% CI: -0.14; 4.56) only in the per protocol analysis. CONCLUSION This study suggests improvement of verbal learning ability and memory of children when supplemented with a fish-flour spread rich in n-3 long-chain polyunsaturated fatty acids.

High-DHA eggs: feasibility as a means to enhance circulating DHA in mother and infant.

Dietary DHA enhances infant attention and visual development. Because the DHA content of red blood cells and plasma lipids varies approximately threefold in pregnancy, maternal DHA status may influence subsequent infant function. It would be feasible to study the effects of higher maternal DHA intake on infant development if dietary intake of DHA could be increased by a reliable means. This study was designed to determine whether women provided with one dozen high-DHA hen eggs (135 mg DHA/egg) would consume the eggs and have higher blood DHA levels than women consuming ordinary eggs (18 mg DHA/egg). The study was a randomized, double-masked comparison of the effect of eggs with different concentrations of DHA on intake and blood lipid DHA content of women and their infants. A third nonrandomized group ate few eggs. In this study, DHA intake reported from eggs was eightfold higher in the high-DHA egg group compared to the ordinary egg group. Including all groups, DHA intake ranged from 0 to 284 mg/d. In this intake range, maternal blood lipid DHA content at enrollment best predicted DHA content at delivery, accounting for 36.5 and 51.7% of the variance in ordinary and high-DHA egg intake groups, respectively. The high-DHA vs. ordinary egg groups had similar maternal and cord blood lipid DHA, but there was a positive relationship between maternal plasma phospholipid DHA and daily DHA intake from eggs controlled for study duration (r=0.278, P=0.048). DHA intake and birth weight were also correlated (r=0.299, P=0.041). High-DHA eggs were well accepted and increased DHA intake. Other benefits of DHA intake during pregnancy were also suggested.

Effect of palm oil on plasma lipoprotein concentrations and plasma low-density lipoprotein composition in non-human primates

Palm oil (PO) contains approximately 43% of palmitic acid. It is the most abundant saturated fatty acid in the diet and it is generally considered the primary cholesterol (C)-raising fatty acid. However, the effect of palmitic acid on plasma cholesterol appears to depend on the cholesterol content of the diet. The aim of this study was to determine the effect of PO with either a high-fat, high-C or moderate-fat, moderate-C diet on lipoprotein C and low-density lipoprotein (LDL) composition. Fifty adult, male vervet monkeys were randomly assigned to the high-fat diet group (HFD: 35%E fat, approximately 0.106 mg C/kJ; n = 30) and the moderate-fat diet group (MFD: 30%E fat, approximately 0.027 mg C/kJ; n = 30). Baseline LDL-C, high-density lipoprotein (HDL)-C and body weight were used to stratify the vervets into comparable experimental groups within each dietary group. The HFD group was divided into two groups of 10 each: one group continued with the HFD in which 8.1%E was derived from lard (AF); in the other group, AF was substituted isocalorically with PO. The MFD group was divided into three groups of 10 each: one group continued with the MFD in which 11.8%E was derived from AF; in the other two groups, the AF was substituted isocalorically with either sunflower oil (SO) or PO. This article presents preliminary results on plasma lipoproteins and LDL composition after 6 months of dietary intervention. Plasma total and LDL-C was higher in all the groups, but the mean changes elicited by PO with either the HFD or MFD were no different from that observed with AF and SO. There was no difference in the mean change of LDL molecular weight within the HFD and MFD. It is concluded that PO is no different from AF (HFD and MFD) or SO (MFD) in its cholesterolaemic effect.

The influence of fish oil supplementation on plasma lipoproteins and arterial lipids in Vervet monkeys with established atherosclerosis

There is controversy about whether supplementing diets with marine fish oil can regress, promote or prevent atherosclerosis. Therefore the effects of an Atlantic pilchard oil (FO) supplement and dietary change were measured in a proven atherosclerosis model. Vervet or African Green monkeys were fed an atherogenic diet (AD) for long enough to ensure progression before treatments started. Matched groups were then treated for 20 months, either by adding FO to the AD (AD/FO), or by changing to a therapeutic diet with FO (TD/FO). Control treatments consisted of supplementing with sunflower oil (SO) instead of FO, so that treatments were AD/SO and TD/SO. The same total polyunsaturates were supplied by the FO and SO and the dose of FO was realistic (2.5% of total energy). A reference group (R) received the TD with no oil supplements. Supplementing with FO did not change the concentrations of total, low or high density lipoprotein cholesterol in plasma. After The AD/FO the intimas of aortas contained more total (p < or = 0.001), free (p < or = 0.05) and esterified (p < or = 0.05) cholesterol, total phospholipid (p < or = 0.01) and sphingomyelin (p < or = 0.05) than after the AD/SO. After FO supplementation eicosapentaenoic acid was significantly higher and arachidonic acid significantly lower in the plasma and aorta intima phosphatidylcholine. None of these changes was anti-atherogenic in terms of atherosclerosis measured in the same individuals (1). Nor did FO increase the efficacy of the TD.

The effect of iron fortification on the fatty acid composition of plasma and erythrocyte membranes in primary school children with and without iron-deficiency

An intervention study was designed to evaluate the fatty acid (FA) status of children aged 6-11 years before and after iron fortification. Iron-deficient (ID) and matched controls without ID (n = 30) were selected. All children received soup (160 ml) fortified with 20 mg iron and 100 mg vitamin C for 15 weeks on school days. Measurements before and after intervention included dietary intake, haematological and iron status and FA composition of plasma and erythrocyte membranes (EMBs). The prevalence of low plasma ferritin concentration and transferrin saturation decreased in the ID children by 40% and 56%, respectively, with intervention. Plasma FAs reflected dietary FA intake. In comparison with controls, the ID group presented with increased percentage total saturated FAs (SFAs; p = 0.0002) in their EMB phosphatidylcholine (PC) and reduced percentage total polyunsaturated FAs (PUFAs; p = 0.0037) before intervention. Lower total n-3 FAs (p = 0.0070), including eicosapentenoic acid (EPA; p = 0.0034), docosapentenoic acid (DPA; p = 0.0048) and docosahexenoic acid (DHA; p = 0.0058), were observed in the ID group. The EMB phosphatidylethanol-amine (PEA) of the ID children presented with lower percentages of alpha-linolenic acid (ALA; p = 0.0001), EPA (p = 0.0051) and DHA (p = 0.0084) compared to controls before intervention. Iron intervention was associated with an increase (p < 0.05) in the percentage of n-3 FAs in the EMB-PC and EMB-PEA of the ID group to percentages comparable to that in the control group. It appears that iron status can influence FA metabolism of specific n-3 FAs in the EMBs of young children.

Long-term evaluation of a micronutrient-fortified biscuit used for addressing micronutrient deficiencies in primary school children.

OBJECTIVE To evaluate the long-term effect on micronutrient status of a beta-carotene-, iron- and iodine-fortified biscuit given to primary school children as school feeding. DESIGN Children receiving the fortified biscuit were followed in a longitudinal study for 2.5 years (n = 108); in addition, cross-sectional data from three subsequent surveys conducted in the same school are reported. SETTING A rural community in KwaZulu-Natal, South Africa. SUBJECTS Children aged 6-11 years attending the primary school where the biscuit was distributed. RESULTS There was a significant improvement in serum retinol, serum ferritin, haemoglobin, transferrin saturation and urinary iodine during the first 12 months of the biscuit intervention. However, when the school reopened after the summer holidays, all variables, except urinary iodine, returned to pre-intervention levels. Serum retinol increased again during the next 9 months, but was significantly lower in a subsequent cross-sectional survey carried out directly after the summer holidays; this pattern was repeated in two further cross-sectional surveys. Haemoglobin gradually deteriorated at each subsequent assessment, as did serum ferritin (apart from a slight increase at the 42-month assessment at the end of the school year). CONCLUSIONS This study has shown that fortification of a biscuit with beta-carotene at a level of 50% of the Recommended Dietary Allowance (RDA) was enough to maintain serum retinol concentrations from day to day, but not enough to sustain levels during the long school holiday break. Other long-term solutions, such as local food production programmes combined with nutrition education, should also be examined. The choice of the iron compound used as fortificant in the biscuit needs further investigation.

The effect of dietary iron deficiency on the fatty acid composition of plasma and erythrocyte membrane phospholipids in the rat

Severe iron deficiency was introduced in rats by feeding outbred male Wistar rats a purified diet that was either adequate or deficient in iron. The rats were weighed regularly over 4 weeks to monitor body weight differences, after which blood was drawn from a subsample to determine the haemoglobin concentrations and fatty acid composition of plasma total phospholipids and to measure the erythrocyte membrane phosphatidylcholine and phosphatidylethanolamine levels. Comparisons between dietary iron adequate (control) and dietary iron deficient (experimental) rats showed that the experimental rats had lower body weight and plasma total phospholipid linoleic acid levels typical of the symptoms of essential fatty acid deficiency. Erythrocyte membrane phosphatidylethanolamine arachidonic acid levels were increased (P < 0.05) with concomitant decreases in oleic acid (P < 0.01). Correlations between fatty acids and growth suggest that the mechanism whereby iron deficiency affects growth is in some way related to abnormal fatty acid shifts that disturb the delicate balance of essential fatty acids in membranes. Additional omega 3 and omega 6 fatty acids may be necessary to counteract the effect of iron deficiency in rats.

Dietary modulation of fatty acid profiles and oxidative status of rat hepatocyte nodules: Effect of different n−6/n−3 fatty acid ratios

Male Fischer rats were fed the AIN76A diet containing varying n−6/n−3 FA ratios using sunflower oil (SFO), soybean oil (SOY), and SFO supplemented with EPA-50 and GLA-80 (GLA) as fat sources. Hepatocyte nodules, induced using diethylnitrosamine followed by 2-acetylaminofluorene/partial hepatoctomy promotion, were harvested, with surrounding and respective dietary control tissues, 3 mon after partial hepatectomy. The altered growth pattern of hepatocyte nodules in rats fed SFO is associated with a distinct lipid pattern entailing an increased concentration of PE, resulting in increased levels of 20∶4n−6. In addition, there is an accumulation of 18∶1n−9 and 18∶2n−6 and a decrease in the end products of the n−3 metabolic pathway in PC, suggesting a dysfunctional Δ-6-desaturase enzyme. The hepatocyte nodules of the SFO-fed rats exhibited a significantly reduced lipid peroxidation level that was associated with an increaser in the glutathione (GSH) concentration. The low n−6/n−3 FA ratio diets significantly decreased 20∶4n−6 in PC and PE phospholipid fractions with a concomitant increase in 20∶5n−3, 22∶5n−3, and 22∶6n−3. The resultant changes in the 20∶4/20∶5 FA ratio and the 20∶3n−6 FA level in the case of the GLA diet suggest a reduction of prostaglandin synthesis of the 2-series. The GLA diet also counteracted the increased level of 20∶4n−6 in PE by equalizing the nodule/surrounding ratio. The low n−6/n−3 ratio diets significantly increased lipid peroxidation levels in hepatocyte nodules, mimicking the level in the surrounding and control tissue while GSH was decreased. An increase in n−3 FA levels and oxidative status resulted in a reduction in the number of glutathione-S-transferase positive foci in the liver of the GLA-fed rats. Modulation of cancer development with low n−6/n−3 ratio diets containing specific dietary FA could be a promising tool in cancer intervention in the liver.

Lipids and Δ6-desaturase activity alterations in rat liver microsomal membranes induced by fumonisin B1

Alterations in the membrane structure and function of hepatocyte membranes by fumonisin B1 (FB1) have been proposed to play an important role in the disruption of growth regulatory effects and hence in the cancer-promoting ability of the mycotoxin. Detailed analyses of lipids in liver microsomal fractions of rats exposed to different dietary levels of FB1 over a period of 21 d indicated an increase in PC, PE, PI, and cholesterol (Chol). These changes decreased the PC/PE and increased the total phospholipid/Chol ratios. When considering FA content, the quantities of total FA increased (P<0.05) in the major phospholipid fractions as a result of the increased phospholipid levels. However, when considering the relative levels (mg/100 mg of the total FA) of specific FA, the monounsaturated FA (16∶1n−7 and 18∶1n−9) and 18∶2n−6 increased (P<0.05), whereas the long-chain PUFA decreased (P<0.05) in the main phospholipid fractions. Enzyme analyses indicated that the activity of the Δ6-desaturase was significantly reduced in liver microsomal preparations in a dose-dependent manner. An increase in the 20∶3n−6/20∶4n−6 ratio also suggested a decrease in the activity of the Δ5-desaturase. Disruption of microsomal lipid metabolism at different levels by FB1 could play an important role in the alteration of growth regulatory effects in the liver.

Thresholds and kinetics of fatty acid replacement in different cellular compartments in rat liver as a function of dietary n-6/n-3 fatty acid content

The kinetics of fatty acid (FA) replacement in different membrane compartments in the rat liver were investigated using diets with varying n-6/n-3 FA ratios. Rats at different stages of growth, i.e. after weaning and at 150 g body weight, were either fed a modified AIN 76A diet containing sunflower oil as fat source or the same diet containing sunflower oil and fish oil to achieve n-6/n-3 FA ratios of 12:1 and 6:1 (diets A and B, respectively). In the adult rats, fed diet A for 8 weeks, C18:2n-6 increased significantly at week 2 in the phosphatidylcholine (PC) fraction of the plasma membranes, microsomes and plasma but not in phosphatidylethanolamine (PE), C20:3n-6 increased significantly at week 2 in the plasma membrane and microsomal PC, but did only increased in PE of both compartments by week 8. C20:4n-6 and the n-3 FAs significantly decreased and increased, respectively, at week 2 in PC and PE of both membrane compartments and plasma PC. The experimental diets led to a change in the plasma membrane fluidity but not in the microsomes. The FA changes in the weaned rats followed a similar pattern as in the adult rats although the changes were greater, depending on the phospholipid fraction and specific FA. The decrease in C20:4n-6 was significantly greater in the microsomal PC and PE and plasma PC but not in the plasma membrane PC and PE. The n-3 FAs increased significantly above the adult levels in the plasma membrane PC and PE respectively but not in the microsomal phospholipid fractions. A plateau for maximal n-3 and n-6 FA incorporation was achieved in the adult rats fed diet A in the microsomes after 2 weeks with no further alterations occurring with diet B. In the plasma PC and plasma membranes most of the n-3 FAs achieved a threshold incorporation after 2 weeks on diet A, except for C22:6n-3 in the plasma membranal PE and certain n-6 FAs in the plasma membrane PC and PE. The present data shows that differences exist in the kinetics of FA incorporation and replacement depending on the specific phospholipid fraction, membrane compartment, age and to a certain extent the dietary n-6/n-3 FA ratio.