Lipid oxidation is the main factor which limits the shelf-life of meat when held under frozen storage. Research undertaken used pork phospholipid liposomes as a model for studying lipid oxidation in meat. Oxidation was followed by monitoring the decrease in the phospholipid unsaturated fatty acyl chains. It was found that the greater the level of unsaturation of the phospholipid fatty acyl chain the greater was their susceptibility to peroxidation. However, the results were not consistent and several reasons for the variation in rate are provided. At ambient temperatures copper (II) was found to be pro-oxidant in the peroxidation of liposomes. At temperatures below 0°C the prooxidant activity of copper (II) was significantly reduced. However copper again became highly pro-oxidant if sodium chloride was present. It is suggested that salt controls the copper ion concentration at sub-zero temperatures as the pro-oxidant activity of copper (II) is reduced on increasing the copper (II) concentration from 0.9 to 90 ppm. Other experiments found sodium nitrite and pholyphosphate to act as antioxidant and that liposome structure was an important factor in the rate of peroxidation. Four storage trials on pork burgers were undertaken to determine whether salt was also pro-oxidant in the stability of pork when held under frozen storage. The oxidative deterioration of the meat was followed by the following methods of analysis:- 1. The decrease in the unsaturated acyl chains of both total lipid and phospholipid. 2. The change in the colour parameters of the meat using reflectance spectroscopy. 3. The analysis of neutral lipid oxidation products by HPLC. 4. The organoleptic qualities of the pork using a trained panel of food assessors. The results from these storage trails showed that the deterioration of pork was minimised by storing the burgers at lower temperatures within the range 0 to -30°C. Salt was found to accelerate the oxidative deterioration of both uncooked and cooked pork when stored at -20°C. Nitrite was found to exhibit some antioxidant behaviour and reduce the pro-oxidant effect of salt.

This work investigated the consequences of vegetable oils spills in salt marsh sediments. The role of autochthonous bacteria in the oils degradation and degradative pathways were also studied 'in situ' and 'in vitro'. Simulated spills of sunflower and linseed oils revealed that both oils penetrated the sediments at a rate of 10-7 CM2 s-1. However, whereas 60% of the linseed oil had disappeared from the sediments after 2 months most of the sunflower oil remained after 6 months. Differences were noted in the adsorption of the oils to sediment particles and the depth at which they accumulate and these factors most likely influenced the route of the oil degradation and the sediments properties such as permeability. The contamination of the sediments with vegetable oils lead to a noticeable reduction in the abundance of plant roots and infauna. The abundance of aerobic, anaerobic and sulphate reducing bacteria in the sediments was increased by the addition of both oils, with linseed oil supporting greater bacterial density than sunflower oil. During the course of the experiment the relative abundance of oil degrading bacteria also increased. As a consequence of the increased bacterial activity, the sediments pH and Eh decreased and anoxic conditions were established, earlier in the case of linseed than that of sunflower oils. The degradation of the oils appeared to be a sequential process, initiated by the aerobic and/or anaerobic bacteria and continued by the sulphate reducing bacteria which themselves where unable to utilise the raw oils. The original composition of both oils underwent alterations mostly associated with their main fatty acid: the concentration of 18: 3(o3 and 18: 2o)6 in linseed and sunflower oil, respectively, decreased whereas that of the remaining fatty acids increased. As a result of the bacterial degradation of the vegetable oils 'new' fatty acids were detected and their identification was attempted using GC-MS analysis of their picolinyl and methyl esters. Various degradative pathways of linseed and sunflower oils involving the formation of the 'new' fatty acids are suggested with isomerisation, hydrogenation and P-oxidation as the primary routes for the degradation.

Orientador: Daniel Barrera Arellano / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-25T11:28:26Z (GMT). No. of bitstreams: 1 Chaves_KamilaFerreira_M.pdf: 1834021 bytes, checksum: 17ab04d365f81e6b3faf3237da71b9f0 (MD5) Previous issue date: 2014 / Resumo: A redução do consumo de ácidos graxos saturados (AGS) na dieta, a fim de reduzir o risco de doenças cardiovasculares, tornou indispensável a busca por substitutos desse tipo de gordura, mediante novas alternativas tecnológicas, como a dos organogéis. O uso de estruturantes em sistemas lipídicos é visto como uma alternativa promissora, pois altera as propriedades físicas de óleos vegetais, sem que ocorram modificações químicas e aumento do teor de AGS, abrindo novas possibilidades para a produção de margarinas e spreads low sat e livres de ácidos graxos trans. O objetivo desse trabalho foi produzir margarinas utilizando a tecnologia de estruturação de óleos para reduzir a quantidade de AGS em comparação às margarinas comerciais convencionais. As matérias-primas, óleo de girassol alto oleico (OGAO), óleo de soja (OS), gordura interesterificada (GI), cera de candelilla (CC) e monoacilgliceróis (M), foram caracterizadas quanto a composição, qualidade e propriedades térmicas. O processo de elaboração das margarinas teste foi realizado em escala de bancada (1 kg), a fase lipídica foi fundida a 80ºC e misturada com a fase aquosa a uma temperatura de 60ºC, sob agitação de 300 rpm. O resfriamento até 30ºC ocorreu sob agitação a 37 rpm em uma sorveteira elétrica com parede de isolamento duplo, previamente resfriada em estufa B.O.D. a 5ºC. As margarinas comerciais avaliadas apresentaram composição de AGS entre 20,52 e 23,93% e teores de lipídios de 70 a 82%. A redução do teor de AGS foi calculada considerando a diferença da quantidade de AGS das margarinas teste em comparação com a média das margarinas comerciais. Foi possível notar através do planejamento experimental que quanto menores os teores de GI, CC e M nas formulações, maior a redução de AGS. As margarinas teste apresentaram melhor estabilidade da emulsão para as temperaturas avaliadas em comparação com as margarinas comerciais. Ocorreu maior exsudação de óleo nas margarinas comerciais em comparação às margarinas desenvolvidas conforme o planejamento experimental na temperatura de 35ºC por 48 horas. As margarinas testadas com OGAO e OS apresentaram menor valor na luminosidade (*L) em comparação às margarinas comerciais. A consistência e a espalhabilidade das margarinas testadas foram significativamente iguais a pelo menos 2/3 das margarinas comerciais avaliadas e a dureza foi estatisticamente igual para todas as margarinas testadas quando comparadas com as margarinas comerciais. Conclui-se que é possível produzir margarinas, mediante a tecnologia de estruturação de óleos, utilizando CC, M e GI como estruturantes, com redução do teor de AGS de 17,3 a 36,6% para as margarinas produzidas com OS e OGAO, respectivamente, em comparação com margarinas comerciais / Abstract: Reduce the consumption of saturated fatty acids (SFA) in the diet in order to reduce the risk of cardiovascular diseases, has made indispensable the search for substitutes for this kind of fat, through new technological alternatives, such as organogels. The use of structuring agents in lipid systems has been presented as a promising alternative, once that they change physical properties without any chemical modification or increase on the amount of SFA, which open new possibilities to the production of low sat and trans free margarines and spreads. The aim of this study was to produce margarines using oil structuring technology in order to reduce the amount of SFA compared to commercial margarines. Raw materials such as high oleic sunflower oil (HOSO), soybean oil (SO), interesterified fat (IF), candelilla wax (CW) and monoacylglycerols (M), were characterized due to their quality parameters, chemical composition and thermal properties. The processing of margarines were conducted in workbench scale (1kg per batch), the oil phase was melted, heating up to 80°C, and then mixed with an aqueous phase at 60°C, under controlled speed at 300 RPM. Cooling up to 30°C occurred under shear at 37 rpm using an electrical ice cream machine with a double wall isolated cube, previously cooled to 5°C. Commercial margarines with a SFA content between 20.52 and 23.93% and 70 to 82% amount of lipids were used as comparative standars. The reduction of SFA was calculated considering the amount of SFA on trial margarines and commercial comparative standards. It was possible to notice, through an experimental design, that smaller amounts of IF, CW and M on the formulations, resulted on reduction of SFA. Trial margarines presented better emulsion stability that commercial products. A higher oil exudation occurred for commercial margarines compared to the trial samples at 35°C for 48 hours. Tested margarines using HOSO and SO presents lower luminosity values (*L) when compared to commercial ones. Consistency and spreadability of trial margarines were statistically equal to, at least, 2/3 of the commercial margarines evaluated and hardness presented no statistical difference among all trial margarines when compared to commercial margarines. In conclusion, margarines produced by oil structuring technology using CW, M and IF as structuring agents, reduced the amount of SFA from 17.3 up to 36.6% in trial margarines with SO and HOSO, respectively when compared to commercial margarines / Mestrado / Tecnologia de Alimentos / Mestra em Tecnologia de Alimentos

Abstract Maintenance of the mitochondrial respiratory chain complexes plays crucial role for the aerobic metabolism of the eukaryotes such as unicellular yeasts, for example, Saccharomyces cerevisiae as well as of human being. Mitochondrial respiratory function has been studied using the yeast S. cerevisiae as a model organism. Since yeast cells are also able to grow without respiration by fermentation, identification of the nuclear genes linked to respiratory function is possible by generation of nuclear gene deletions and testing for respiration-deficient phenotype of the yeast deletion strains id est for yeast cells only poorly or not at all growing on the media containing non-fermentable carbon sources. This study reports identification of a novel mitochondrial 2-enoyl thioester reductase from the yeasts Candida tropicalis and S. cerevisiae, Etr1p and Mrf1p, respectively. Examination of the function of these proteins in the respiration-deficient mrf1Δ strain from S. cerevisiae suggests that the reductase is involved in mitochondrial fatty acid synthesis (FAS type II) in the yeast. Site-directed mutagenesis of a conserved tyrosine in the catalytic site of the enzyme indicated that the 2-enoyl thioester reductase activity is critical for mitochondrial respiratory competence. In addition, subcellular localization to mitochondria was required for the complementation of the respiration-deficient phenotype of the yeast reductase deletion strain. The crystal structure for the Etr catalytic site mutant indicated the structural integrity of the mutant supporting the requirement of the tyrosine for the catalysis. Characterization of Etr crystal structures both in apo- and holo-forms containing NADPH established Etr as a member of novel subfamily of enoyl thioester reductases in the superfamily of medium-chain dehydrogenases/reductases (MDR). Two isoforms of Etr with the difference in three amino acids only are encoded by two distinct genes in C. tropicalis, whereas only single gene encodes the reductase functioning in the mitochondria in S. cerevisiae. The presence of two genes in C. tropicalis was taken as an example of genetic redundancy in this yeast, the two genes also shown to be expressed in slightly different ways under various carbon sources available for growth.

Master of Science / Department of Animal Sciences and Industry / Teresa L. Douthit / Supplementation of lipids in the form of triglycerides is a safe method for increasing energy density in equine diets; however the effects of calcium salts of fatty acids (CSFA) on nutrient digestibilities in the horse are unknown. Eight cecally cannulated Quarter Horses were utilized in a crossover design in which horses consumed 1.5% BW smooth bromegrass hay and 0.5% BW concentrate supplemented with 4.1% soybean oil (SB) or 4.9% Enertia s/f® (E), a proprietary CSFA, for 28 d. Feces were collected for determination of apparent total tract digestibility of nutrients, and cecal digesta was evaluated for pH and concentrations of VFA and LCFA. Serum was collected following a 16-h fast and analyzed for triglycerides and cholesterol. Apparent total tract digestibilities of DM, NDF, ADF, CP, ether extract, and GE were unaffected by lipid source (P > 0.10). Serum triglycerides tended to be greater in horses consuming E compared to SB (P = 0.10); but, serum cholesterol concentrations were not different (P = 0.45). Cecal pH was unaffected by lipid source. Cecal concentration of total VFA tended to be greater in horses consuming SB compared to E at 2-h post feeding (P = 0.07). Cecal propionate concentrations were greater at 2-h post feeding for horses fed SB compared to horses fed E (P = 0.03). A treatment × time interaction was detected for total cecal LCFA concentration in which total cecal LCFA were greater in horses consuming E compared to SB at 2-h following consumption of a meal but became more similar as time progressed (P < 0.01). Secondly, an in vitro study was conducted to determine the effect of Co chloride on fermentation parameters using an equine cecal fluid inoculum in a 2 × 5 factorial arrangement of treatments. Alfalfa or smooth bromegrass hay (5 g DM) were utilized as substrates and were supplemented with 0.0, 0.5, 5.0, 25.0, or 50.0 mg Co/kg substrate DM. Cultures containing alfalfa had greater IVDMD compared to those with smooth bromegrass hay (P < 0.01). There was a forage × Co interaction in which gas production increased with time and was greater in cultures containing alfalfa (P < 0.01). Production of acetate, propionate, butyrate, isobutyrate, isovalerate, and total VFA, as well as acetate:propionate, were increased in cultures containing alfalfa compared to those containing brome hay (P < 0.05).

Abstract The semi-domesticated reindeer is a northern species of Cervidae that is exposed to extreme seasonal changes in temperature and nutrition in its living environment. The objective of this study was to examine the significance of thermogenic brown adipose tissue (BAT) for the survival of newborn reindeer in the cold during the critical perinatal period. The other main objective was to study the effect of wintertime undernutrition on serum and bone marrow fatty acid composition in yearling reindeer, with particular attention on the proportions of unsaturated and polyunsaturated fatty acids (PUFAs) and their feasibility as indicators of nutritional status. The results showed that the most of the adipose tissues in newborn reindeer were functionally active BAT. The tissue had specific anatomical locations, specialized cell morphological structure, high aerobic capacity, and tissue-specific mitochondrial 32000 Mr-uncoupling protein (UCP1) that is considered a rate-limiting factor for thermogenesis. The most readily mobilized fatty acids from BAT triacylglycerols were arachidonic, linoleic, and α-linolenic acids. BAT was most active at birth and during the close perinatal period but its aerobic capacity declined during the first month of life while UCP1 disappeared and the tissue gradually adopted the histological characteristics of white adipose tissue. The newborn reindeer had very low proportions of the principal C18-PUFAs, linoleic and α-linolenic acids, in serum lipids. However, the proportions of C18-PUFAs increased during the first few days of life by a rate that suggests a selective retention of these fatty acids from milk lipids. A prolonged restricted feeding of reindeer with lichen during winter and spring induced significant reductions in the proportions of linoleic and α-linolenic acids in serum cholesteryl esters and phospholipids, while proportion of arachidonic acid and serum prostaglandin PGF2α metabolite concentration increased. Plasma leptin and insulin levels decreased in parallel with decreases in feed intake and body weight. In freely ranging reindeer, the proportions of oleic acid and principal C18-PUFAs were significantly reduced in femur bone marrow triacylglycerols as a result of a wintertime undernutrition. In conclusion, active BAT is the dominant adipose tissue type in the newborn reindeer and it is likely to have a major significance on the thermoregulatory heat production and cold resistance of reindeer during the perinatal period. The changes in the specific PUFA proportions of serum and bone marrow lipids refleict the changes in the nutritional status and suggest that these fatty acids are preferentially utilized during prolonged wintertime undernutrition.

This thesis describes an attempt to increase the life span of mice after injection of a leukemic tumor. Fatty acids were used as a protecting agent against the leukemic tumor.

Background and Aim: Microalgae are a potential economical source of omega-3 fatty acids, which are known for their health benefits. Omega-3 fatty acids derived from microalgae are however still expensive due to high costs involved in production. These costs can partly be attributed to cultivation of microalgae in unialgal cultures, which are prone to contamination, difficult to scale up and require stringent control of growth conditions. This study therefore is aimed to investigate the feasibility of using a mixed microalgae culture for the production of omega-3 fatty acids. Methods and materials: The microalgae were grown under uncontrolled conditions in a hanging-bag photo-bioreactor, with only the phosphorus-nitrogen ratio (P:N-ratio) varied. Variations in the different population proportions in the culture were studied and related to the biomass, total fatty acid and omega-3 fatty acid productivity of the culture. Finally, a comparison was made between the results obtained in this study and similar studies found in literature. Results and discussion: The mixed culture used was composed of several green microalgae, whose proportions varied with P:N-ratio, season, and age of the culture. Different interactions between the populations were observable, and these were thought to influence the culture’s biomass, total fatty acids (TFAs) and omega-3 fatty acid productivity. The TFA content of the culture was ~10-20% by the end of the exponential phase, depending on the nutrients supplied. The fatty acids were composed of mostly unsaturated fatty acids, the majority of which were omega-3 fatty acids; C18:3ω3 (up to 50% of TFA), C16:4ω3 (up to 15%) and C18:4ω3 (up to 5%). Other fatty acids observed included C16:0 (up to 25%), C18:1ω9/C18:1ω8 (up to 20%), C18:2ω6 (up to 5%), traces of C18:0, C18:3ω6, C17:0, C16:3ω3 and C16:1ω7 were also detected. The presence of these fatty acids was found to vary depending on season, P:N-ratio, and age of the culture. When compared to studies in the literature, the mixed culture was found to perform better than, or as well as reported studies, where unialgal cultures were cultivated under controlled laboratory conditions. Conclusion: In view of the reported advantages of cultivating mixed cultures, the production of omega-3 fatty acids via such cultures appears attractive. More research however is needed to optimise the culture; study interactions between the organisms and their effect on biomass, total fatty acid and omega-3 fatty acid productivity. Finally, the economic implications of using mixed cultures need to be evaluated as well.

Microbiota are present in large numbers and as a diverse population within the gastrointestinal tract. There are approximately 400 different species of microbiota which may be beneficial, harmful or both, but each play an important role in the regulation and modulation of the hosts’ bowel processes (McOrist et al. 2008; Dethlefsen et al. 2008). Many of these colon microbiota allow for saccharolytic fermentation of non-digestible dietary fibres and carbohydrates into by-products and intermediates, followed by a subsequent conversion into short chain fatty acids (SCFAs) (mainly n-butyric acid, propionic acid and acetic acid) each of which play an important role in maintaining colon homeostasis (Topping & Clifton 2001). A balance of ‘good’ microbiota (e.g., Bacteroides spp./ Bifidobacteria spp.) and ‘bad’ microbiota (e.g., Veilonellae) and the optimal production of various SCFAs within the gut could possibly allow for proper functioning of the large intestine and assist in decreasing the onset of various colonic disorders such as Irritable Bowel Syndrome (IBS). The sample group for the study consists of male and female patients, with an average age of 40 to 50 years old, whom of which have been diagnosed with either constipation IBS (C-IBS) or diarrhoea IBS (D-IBS) via the Rome III criteria system for IBS diagnosis. DNA and SCFA extractions were optimised for human stool, colonic fluid and tissue biopsy sample obtained from the aforementioned patients. Optimization steps allowed for starting material with high analysis integrity. Different methods of microbiota analysis, such as ARISA, were investigated; however, real-time qPCR was selected as the best method to identify and quantify specific microbiota. Extracted SCFAs were separated via gas chromatography and identified and quantified via Mass Spectrometry. Significant changes in microbial content and SCFA profiles were found to be associated with healthy and IBS patients. Results obtained would however be influenced by external factors typical of clinical studies of this nature. This study allows for opportunities for future research into understanding IBS.

Docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6) are deposited in large amounts in membrane phospholipids of the developing central nervous system (CNS). High levels of 22:6n-3 are found in synaptic terminals and retina, and are important for normal visual development and function. 20:4n-6 and22:6n-3 are supplied in human milk. In contrast, infants fed formula rely completely on endogenous synthesis of 20:4n-6 and 22:6n-3 from linoleic (18:2n-6) and a-linolenic (18:3n-3) acid, respectively. Levels of 22:6n-3 in the blood lipids of infants fed formula are lower than in infants fed human milk. Concern over the supply of 22:6n-3 led to clinical trials in which premature infants were fed formulas containing fish oils as a source of 22:6n-3. Piglets, which have a similar lipid metabolism and perinatal timing of the brain growth spurt to humans, have a lower percentage of 22:6n-3 in blood, liver and CNS tissues when fed formula with 30% of fatty acids as18:2n-6 and 0.8% 18:3n-3, compared to sow milk. It was hypothesized that the low blood and tissue 22:6n-3 in formula-fed piglets was due to inappropriate quantities and/or ratios of dietary 18:2n-6 and 18:3n-3 limiting the synthesis of 22:6n-3. Thus, the main objectives of this thesis were to determine. (1) if 22:6n-3 is an essential dietary nutrient for the term gestation piglet, (2) if appropriate quantities and ratios of 18:2n-6 and 18:3n-3 in formula will support CNS membrane accretion of 20:4n-6 and 22:6n-3, comparable to piglets fed varying amounts of 22:6n-3 in natural milk, and (3) if lower blood phospholipid 22:6n-3 consistently reflects reduced 22:6n-3 in the CNS. Initial studies (Experiment I) showed that formula with 4% 18:3n-3 supported a similar percentage of22:6n-3 in piglet liver and CNS membrane lipids to sow milk, but was associated with lower brain weight. Deposition of 22:6n-3 in brain was influenced by the formula 18:3n-3 content. The 18:2n-6:18:3n-3 ratio (22:1and 37:1) seemed to be important, however, when formulas contained 1% 18:3n-3. Low levels of fish oil in formula, similar to those used in clinical trials, were effective in supplying 22:6n-3 to the developing piglet brain (Experiment II). The efficacy of 18:3n-3 in supporting the deposition of 22:6n-3 in the brain was estimated to be at least 20% that of dietary 20:5n-3 plus 22:6n-3. With increasing dietary fish oil, however, levels of eicosapentaenoic acid (20:5n-3) increased and 20:4n-6decreased in plasma, liver and retina, but not brain (Experiment III). This suggests regulatory mechanisms may exist to maintain relatively constant levels of 20:4n-6 and 20:5n-3 in brain. Milk 22:6n-3 varies with maternal intake of 22:6n-3. The effect of milk 22:6n-3 content was studied in piglets fed milk with 0.1% or 1.5% 22:6n-3 obtained from sows fed usual pig diets containing vegetable fats without or with fish oil, respectively (Experiment IV). Consumption of 1.5 vs 0.1% 22:6n-3 from sow milk resulted in 300% higher 22:6n-3 in liver and blood phospholipids and 11% higher 22:6n-3 in cerebrum of nursing piglets. Despite similar milk 20:4n-6, the % 20:4n-6 in tissues other than the brain was lower in piglets fed high22:6n-3 sow milk. Thus, high intakes of n-3 fatty acids decrease 20:4n-6 in piglet liver and blood lipids. The blood phospholipid % 22:6n-3 in piglets fed formulas containing 18:2n-6 and 18:3n-3 but not their long-chain derivatives, was lower than in piglets fed 22:6n-3 in natural milk, consistent with published findings in formula-fed infants. However, in contrast to circulating lipids, formulas with 4% 18:3n-3 maintained similar levels of 22:6n-3in the piglet CNS compared to milk. These studies show that blood phospholipid 22:6n-3 and 20:4n-6 are not specific indices of effects in CNS lipids. This thesis has shown (1) 22:6n-3 is not essential in the diet of the term piglet, if adequate 18:3n-3 is given, (2) fish oils are an effective source of 22:6n-3 for deposition in the developing brain, (3) high dietary n-3fatty acids interfere with 20:4n-6 metabolism, and (4) blood lipid 20:4n-6 and 22:6n-3 do not accurately reflect CNS fatty acids. / Land and Food Systems, Faculty of / Graduate