Heat exposure causes several physiological and metabolic alterations. Although lipids are vital in sustaining energy production, heat-induced alterations in lipid metabolism have not been clearly established. CHAPTER 1 reviews the known metabolic alterations resulting from heat stress, with a specific focus on changes in whole-body lipid utilization and plasma lipids. CHAPTER 1also outlines the physiological changes caused by heat stress, and their role in reducing exercise performance. The study presented in CHAPTER 2 has shown that, compared to thermoneutral conditions, NEFA concentrations were 37% higher following passive heating and 34% higher following exercise in the heat, without significant changes in whole-body lipid utilization. In addition, the level of hyperthermia attained during passive pre-heating and exercise in the heat resulted in a 13% decrease in total external work and a significantly higher rate of perceived exertion. CHAPTER 3 summarizes the study results and presents the limitations and applications of the study.

Current trends toward warmer air temperatures and longer ice free seasons in Hudson Bay are expected to cause changes in Arctic marine ecosystem dynamics. Ringed seals (Phoca hispida) will likely experience changes in levels of predation, competition, and prey availability. The purpose of this thesis was to investigate seasonal, inter-annual, and spatial variation in Hudson Bay ringed seal feeding ecology. Fatty acid composition, δ15N, and δ13C varied significantly by season, suggesting seasonal changes in foraging habitat and diet. Spatial differences in ringed seal stable isotope ratios occurred between western and eastern Hudson Bay, and there was a strong relationship between spring air temperature and δ15N. Peak δ15N occurred within a range in spring air temperatures between approximately -5°C and -2°C. I propose that the high δ15N observed in ringed seals within this temperature range is indicative of relatively greater importance of capelin (Mallotus villosus) in the ringed seal diet.

This thesis presents evidence for a novel mechanism of regulation of pyruvate dehydrogenase (PDH) kinase by fatty acids and also results of a study of muscle triacylglycerol concentration. In animals regulation of PDH complex activity is central to the selection of respiratory fuels and to the conservation of glucose during carbohydrate deprivation. The principal means of regulation of PDH complex is interconversion of phosphorylated (inactive) and dephosphorylated (active) forms effected by PDH kinase and PDH phosphatase. Earlier in vitro studies by others had identified both shorter term (min) and longer term (hours) mechanisms of activation of PDH kinase by fatty acid. In the present study PDH kinase activity (as measured by rates of ATP-dependent inactivation of PDH complex in extracts) was shown to be increased when rat heart mitochondria were incubated with palmitoyl-L-carnitine [PC] (and other CoA utilising respiratory substrates). The activation of PDH kinase persisted through removal of respiratory substrate following incubation with CCCP. A comparable effect of PC was also demonstrable in heart mitochondria from 48h-starved rats (i.e. the mechanism may be distinct from that which increases PDH kinase activity in starvation). Rates of ATP-dependent inactivation of PDH complex were also increased when extracts of rat heart mitochondria were incubated with palmitoyl-CoA (PCoA); the increase was comparable with that seen on incubation of intact mitochondria with PC. The PC effect in intact mitochondria and the PCoA effect in mitochondrial extracts may not be identical as PCoA further increased PDH kinase activity in extracts from mitochondria incubated with PC. Rates of incorporation of ³²P from [γ-<sup>32</supP]ATP into PDH complex were unaltered by pnor incubation of mitochondria with PC or by pnor incubation of mitochondrial extracts with PCoA. Three lines of evidence confirmed that the effect of PC to accelerate ATP-dependent inactivation involved phosphorylation of the PDH complex (viz; use of a non-phosphorylatmg ATP analogue; use of known inhibitors of PDH kinase; and use of known activators/inhibitors of PDH phosphatase). Earlier studies had shown that phosphorylation in punfied bovine and porcine PDH complexes is half site (involves only one α-chain in E1 (α2β2) and had suggested that phosphorylation in rat heart complex may be full site (i.e. involves both α-chains). The present study suggests the possibility that elevation of fatty acyl CoA under slaughter house conditions might be a determinant of half site phosphorylation. A method was developed and evaluated for measurement of triacylglycerol in rat soleus muscle strips with the object of investigating factors that may regulate triacylglycerol synthesis in this muscle. This study was abandoned because, although the method was highly reproducible, great variation was found in the triacylglycerol concentration of individual muscles suggesting the possibility of variable contamination with small amounts of adipose tissue.

Mice (C57BL/6J: B6) exposed to maternal dietary restriction (DR) exhibited fetal growth- restriction and as adults develop symptoms of the metabolic syndrome. We aimed to determine the impact of DR on fetal hepatic gluconeogenic pathway and insulin sensitivity in late gestation. Second, we aimed to determine whether a postweaning diet rich in omega-3 fatty acids would alter the development of glucose intolerance, insulin resistance and obesity in DR male offspring. The reduced rate of fetal glycogen synthesis by DR male offspring and altered hepatic gene expression of enzymes involved in insulin signalling and glucose metabolism suggest abnormal fetal development in response to DR that may contribute to the later development of the metabolic syndrome. The postweaning omega-3 diet improved obesity, glucose intolerance and insulin resistance in both DR and control males. These data suggest that nutrition in pregnancy and postnatal life play important roles in determining life-long metabolic health.

The present research examined the hypothesis that omega-3 polyunsaturated fatty acids would increase seizure threshold in rats in vivo, and reduce neuronal excitability in mouse hippocampal slices. Seizure thresholds were measured in rats using the maximal pentylenetetrazol and electrical stimulation seizure tests following α-linolenic acid (ALA) or docosahexaenoic acid administration. ALA raised seizure threshold in the maximal PTZ seizure test, but this effect probably occurred because ALA displaced DHA from liver to the brain. DHA itself was therefore tested in the PTZ and electrical stimulation seizure tests. Direct administration of DHA by subcutaneous injection raised seizure thresholds in the PTZ seizure test, which models tonic-clonic attacks in humans. Dietary enrichment with DHA raised afterdischarge seizure thresholds in the cortex and amygdala, which model simplex and complex partial seizures in humans, although this effect took some time to occur. In vitro, the application of DHA also reduced the incidence of excitatory sharp waves in mouse hippocampal slices. This effect did not appear to be due to either an increase in GABAergic inhibitory tone, nor to a decrease in glutamatergic drive. The fatty acid composition of phospholipids and unesterified fatty acids were measured in the brain following microwave fixation in order to determine whether the effects of DHA on seizure thresholds were due to its de-esterification from the phospholipid membrane. The assay surprisingly revealed that subcutaneous administration of DHA at a dose that raised seizure threshold, increased unesterified arachidonic acid, but not unesterified DHA concentrations during seizures. The results of these studies support the hypothesis that DHA raises seizure threshold in rats, and reduces neuronal excitability in vitro. The effects of DHA on seizure threshold are possibly mediated by the de-esterification of arachidonic acid, which is known to have effects on the voltage-dependent sodium channel.

Mice (C57BL/6J: B6) exposed to maternal dietary restriction (DR) exhibited fetal growth- restriction and as adults develop symptoms of the metabolic syndrome. We aimed to determine the impact of DR on fetal hepatic gluconeogenic pathway and insulin sensitivity in late gestation. Second, we aimed to determine whether a postweaning diet rich in omega-3 fatty acids would alter the development of glucose intolerance, insulin resistance and obesity in DR male offspring. The reduced rate of fetal glycogen synthesis by DR male offspring and altered hepatic gene expression of enzymes involved in insulin signalling and glucose metabolism suggest abnormal fetal development in response to DR that may contribute to the later development of the metabolic syndrome. The postweaning omega-3 diet improved obesity, glucose intolerance and insulin resistance in both DR and control males. These data suggest that nutrition in pregnancy and postnatal life play important roles in determining life-long metabolic health.

Fatty acid profiling provides information on dietary intakes and an understanding of lipid metabolism. High throughput techniques such as fingertip prick (FTP) sampling has gained popularity in recent years as a simplified method for basic research, and could be further used to assess disease risk in the population, and other similar high-throughput techniques have the potential to assist in the monitoring and labeling of fatty acids in the food supply. With the advancement of high-throughput sample analysis techniques, a more complete understanding of storage stability is required as a larger volume of samples are produced with equal amounts of time to analyze them. Energy-assisted analysis techniques have the potential to help ameliorate some of these issues. Presently, FTP blood, whole blood and salmon storage stability is assessed under various storage conditions, and both microwave-assisted direct transesterification and indirect ultrasound-assisted extraction techniques are assessed. It is determined that storage of FTP blood and whole blood samples at -20°C results in significant and nearly complete highly unsaturated fatty acid (HUFA) degradation compared to all other temperatures examined. This degradation is determined to be the result of hemolysis and subsequent iron release from erythrocytes initiating fatty acid peroxidation reactions. Direct transesterification of FTP blood is reduced from as long as three hours to one minute with microwave-assisted energy and fatty acid extraction from ground flaxseed is reduced to 40 minutes from as long as 24 hours without compromising fatty acid profiles. Results of the current study provides insight into the storage stability of food sample and blood samples collected via high-throughput techniques, and provides support for the utilization of further high-throughput energy-assisted analytical methods that can help to minimize the potentially detrimental effects that long-term storage can have on fatty acid profiles.

The objectives were (1) to establish a profile of total fatty acids and cholesterol in amniotic fluid (AF) as well as (2) to determine possible associations between AT fatty acids (micromolar and relative proportion) with gestational age and birth weight. A total of 208 AF samples collected between 12 and 22 weeks of gestation during routine amniocentesis were analyzed using tandem column gas chromatography (GC). Smoking increased AF polyunsaturated fatty acid (PUFAs) levels while developmental stage and storage time decreased AF fatty acid quantities. AF trans fatty acids (TFAs) were negatively associated with both birth outcomes, whereas specific fatty acids including stearic acid (C18:0) and gondoic acid (C20:1n-9) were identified as negative predictors for gestational age and birth weight respectively. This study demonstrated novel relationships between fatty acids and fetal growth and gestational age in early midgestation AF, suggesting a possible role of AF fatty acids in predicting birth outcomes.

Aquaculture feeds, traditionally composed mainly of fishmeal and fish oil, currently represent the largest cost to fish farmers. With aquaculture growing at an average of 8.8% per year and limited supply of fishmeal and fish oil, suitable alternatives must be found. In addition to increasing sustainability and lowering production costs, the use of plant and/or animal ingredients has the potential to lower flesh levels of persistent organic pollutants (POPs) such as polychlorinated biphenyls. Fish oil and to a lesser extent fishmeal, are considered to be the largest source POPs in farmed fish. Using alternative feed ingredients however, can compromise fish growth and the flesh quality of the final product. Lipid sources including flaxseed oil, canola oil, poultry fat and the protein sources canola protein concentrate, soy protein concentrate and poultry by-product meal were examined as alternatives to fish oil and fishmeal in one on-farm field study and one laboratory feeding trial with Atlantic salmon (Salmon salar) and two laboratory feeding trials conducted on sablefish (Anoplopoma fimbria), a relatively new marine aquaculture species. The nutritive value of the alternative ingredients was assessed on the basis of fish growth performance, proximate composition, fatty acid composition and apparent digestibility coefficients. Sensory attributes were evaluated in the sablefish studies while flesh POP levels were determined in both species. The use of alternative dietary lipids showed no negative effects on fish performance. However replacement of fishmeal with plant proteins in some cases, negatively affected fish growth. Flesh levels of persistent organic pollutants were significantly decreased (p<0.05) with the use of alternative dietary lipids, and flesh levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were also depressed. Activated carbon treated anchovy oil and finishing diets were examined in the Atlantic salmon laboratory feeding trial and were effective at lowering flesh POP levels while providing high levels of EPA and DHA. The use of alternative feed ingredients will soon be inevitable in aquaculture feeds. The current research shows alternative lipids and proteins can be incorporated successfully in sablefish and Atlantic salmon feeds with minimal effects on fish growth and quality.

My thesis research has examined the fatty acid composition of serum triacylglycerol, phospholipid and cholesterol ester in 2793 participants who took part in the 1997 National Nutrition Survey - a national population-based survey of New Zealand adolescents and adults aged or [greater than or equal to]̲15 y. Differences in serum fatty acids by sex, age, ethnicity, body mass index and smoking - independent of dietary fat intake - were determined. Serum fatty acids were used as biomarkers of saturated and polyunsaturated fat intake to predict population serum total cholesterol concentrations. The association between n-3 long-chain polyunsaturated fatty acids in serum phospholipid and mental and physical wellbeing, as assessed by the short form-36, was determined. Serum fatty acids have been used as biological markers of fat intake and to predict the risk of disease. The fatty acid composition of serum triacylglycerol, phospholipid and cholesterol ester is subject to alteration by dietary fat but overall, is largely controlled by metabolic enzymes. Non-dietary variables - sex, age, body mass index or cigarette smoking - may influence the activity of these enzymes, which will subsequently alter the fatty acid composition but the extent to which these affect serum fatty acid composition in the general population is poorly understood. Our results showed that the proportion of docosahexaenoic acid in serum phospholipid and cholesterol ester was significantly greater in women by 0.15 and 0.02 mol%, respectively in comparison to men whereas, the proportion of eicosapentaenoic acid was significantly greater in men by 0.08 and 0.1 mol%, respectively, after adjusting for age, ethnicity, body mass index and smoking. A number of differences in the proportion of palmitoleic acid in serum triacylglycerol, phospholipid and cholesterol ester were detected; palmitoleic acid increased across the age categories in women (15-24, 25-44, 45-64 65+ y), was higher in women compared to men, New Zealand Europeans compared to New Zealand Maori and Pacific People, those with a body mass index or [greater than or equal to] 30 kg/m� compared to those with a body mass index < 25 kg/m� and in current smokers in comparison to non-smokers. In women, there was an inverse trend in the proportion of linoleic acid in serum phospholipid and cholesterol ester across the age categories. The proportion of linoleic acid in serum triacylglycerol, phospholipid and cholesterol ester was lower in smokers by 2.19, 1.04 and 0.75 mol%, respectively in comparison to non-smokers. None of these differences could be explained by a difference in dietary fat intake. Consequently, sex appears to affect the metabolism of n-3 long-chain polyunsaturated fatty acids independent of dietary fat intake and metabolic differences associated with age, body mass index and smoking may be at play for a number of other serum fatty acids notably, palmitoleic and linoleic acids. Evidence for a role of dietary fat as a predictor of serum cholesterol concentrations in the general population is conflicting. On one hand, results from cholesterol-lowering dietary intervention trials show unequivocally that decreasing saturated fat intake produces a meaningful reduction in serum cholesterol concentrations. On the other hand, the results of large observational studies show little association between saturated fat intake and cholesterol concentrations. The lack of association in the latter studies may result from errors in dietary assessment and therefore, using serum fatty acids as biomarkers of fat intake may overcome the limitations associated with typical dietary assessment techniques. Participants were divided into quintiles of increasing proportion of serum fatty acids. Each one SD increase in the myristic acid composition of serum cholesterol ester, triacylglycerol and phospholipid was associated with an increase in cholesterol of 0.19, 0.10 and 0.13 mmol/L, respectively after adjusting for confounding variables. The difference in cholesterol concentrations between those categorised into the highest and lowest quintiles of serum cholesterol ester myristate was 0.48 mmol/L. A one SD increase in the linoleic acid composition of serum cholesterol ester, triacylglycerol and phospholipid corresponded to a decrease in cholesterol of 0.07, 0.05 and 0.07 mmol/L, respectively. The difference in cholesterol concentrations between the 1st and 5th quintiles of serum cholesterol linoleate was 0.18 mmol/L. Intake of saturated and polyunsaturated fats, as measured using serum fatty acids, are important determinants of cholesterol concentrations in New Zealanders. It has been hypothesised that a lower intake of n-3 long-chain polyunsaturated fatty acids, largely of marine origin, is implicated in the aetiology of depressive disorder. Results from the majority of observational studies have shown that depressed participants have a lower proportion of eicosapentaenoic or docosahexaenoic acid in phospholipids compared to controls but evidence for an improvement in depressive symptoms after supplementation with n-3 long-chain polyunsaturated fatty acids is conflicting. There is little known about the role that n-3 long-chain polyunsaturated fatty acids may have as predictors of mental wellbeing in the general population. Participants were categorised into quintiles of increasing n-3 long-chain polyunsaturated fatty acids in serum phospholipid. There was no significant trend in self-reported mental wellbeing - the mental component score - across the quintiles of eicosapentaenoic, docosapentaenoic and docosahexaenoic acids or the sum of these three fatty acids after adjusting for confounding variables. There was a significant trend in the mental component score across the quintiles of the ratio of eicosapentaenoic/arachidonic acid; the difference between the highest and the lowest quintile was 6.6 points. There were significant positive trends in self-reported physical health - the physical component score - across the quintiles of eicosapentaenoic and docosapentaenoic acids as well as the ratio of eicosapentaenoic/arachidonic acid ratio; the difference between the 1st and 5th quintiles were 8.6, 6.0 and 8.9 points, respectively. Overall, there appears to be little association between the n-3 long-chain polyunsaturated fatty acid composition of serum phospholipid and self-reported mental health in a population of low fish consumers; however, the proportion of n-3 long-chain polyunsaturated fatty acids may be an important predictor of physical wellbeing in New Zealanders.