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京都大学 / 新制・課程博士 / 博士(農学) / 甲第23253号 / 農博第2460号 / 新制||農||1085(附属図書館) / 学位論文||R3||N5343(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 小川 順, 教授 阪井 康能, 教授 栗原 達夫 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

<p>Of the three components of parenteral macronutrient classes (protein, carbohydrate and lipid), the lipid class is the least understood and the fatty acid distribution in lipid emulsion products has the potential to play a critical role in the development of infant morbidities. Breast milk has long been considered the gold standard for infant nutrition, but when infants cannot tolerate enteral feeds, the use of lipid emulsions cannot be avoided despite their adverse side effects.</p> <p>In this study, fatty acid profiles from five commercially available lipid emulsion products were compared to the profile of breast milk. As well, resulting serum on each of these nutrition sources were compared to the profile of either lipid emulsion (Intralipid) or breast milk. Fatty acid profiles for matched pairs of breast milk from mother and resulting serum from infants were compared as well as the profile of normotriglyceridemic serum samples to hypertriglyceridemic ones. Results indicate that not one lipid emulsion product is like another, nor do their profiles closely resemble breast milk even though they are intended to replace the fat portion of the infant’s natural source of nutrition. Serum was not found to directly reflect the fatty acid profile of the nutrition source administered, as was expected based on literature, highlighting that there is a complex web of pathways between nutrition administration and appearance of fatty acids in the serum. Further research is necessary to define the effect of fatty acid chain length and degree of saturation on these metabolic pathways, as the very essence of interrelations such as these can complicate interpretations of results.</p> / Master of Science in Medical Sciences (MSMS)

<p>Investigating the presence of microbial communities in the Earth's deep terrestrial subsurface and the metabolic processes taking place in these environments provides insight into the some of the ultimate limits for life on Earth, as well as the potential for microbial life to exist within the subsurface of other planetary bodies. This Master's thesis project utilized phospholipid fatty acid (PLFA) analysis, in combination with carbon isotope analyses (δ¹³C and Δ¹⁴C), to explore the presence and activity of microbial communities living within deep terrestrial subsurface fracture water systems and low permeability, deep sedimentary rocks. Deep fracture water systems, ranging from 0.9 to 3.2 km below land surface, were sampled for microbial communities via deep mine boreholes in the Witwatersrand Basin of South Africa. PLFA concentrations revealed low biomass microbial communities, ranging from 2x10¹ to 5x10⁴ cells per mL and the PLFA profiles contained indicators for environmental stressors, including high temperatures and nutrient deprivation. δ¹³C and Δ¹⁴C analyses of PLFAs and potential carbon sources (dissolved inorganic carbon (DIC), dissolved organic carbon (DOC) and methane) identified microbial utilization of methane in some systems and utilization of DIC in others. Evidence for microbial oxidation of methane and chemoautotrophy in these systems is consistent with a self-sustaining deep terrestrial subsurface biosphere that is capable of surviving independent of the photosphere. Viable microbial communities were also identified within deep (334 to 694 m depth) sedimentary rock cores sampled from the Michigan Basin, Canada. PLFA analyses revealed microbial cell densities ranging from 1-3 x 10⁵ cells/mL and identified PLFA indicators for environmental stressors. These results demonstrate the ubiquity of microbial life in the deep terrestrial subsurface and provide insight into microbial carbon sources and cycling in deep microbial systems which may persist in isolation over geologic timescales.</p> / Master of Science (MSc)

Omega-3 polyunsaturated fatty acids (ω3 PUFAs) provide important health benefits, but dietary consumption is low. Supplementing foods with ω3 PUFAs is of interest, but intervention strategies are necessary to preserve the integrity of these unstable compounds. Microencapsulation of ω3 PUFA sources is one means of improving their stability. In this work, ω3 PUFA microcapsules were prepared by spray drying with chitosan and blends of chitosan, high-amylose starch, and pullulan as wall materials. The primary objectives of this research were (1) to evaluate the effect of chitosan type and oil:wall ratio on ω3 PUFA microcapsule properties, (2) to evaluate the effect of blending chitosan with high-amylose starch and pullulan on ω3 PUFA microcapsule properties, and (3) to evaluate the oxidative stability of ω3 PUFA microcapsules by monitoring primary and secondary oxidation products during storage. Microcapsule encapsulation efficiencies (EE) ranged from 63% to 79% with the highest EEs observed for microcapsules prepared from chitosan with higher degree of deacetylation (DD) and lower molecular weight (MW). Median microcapsule size ranged from 3 μm to 11 μm. Moisture contents were all below 7% and water activities (a_w) were below 0.27. Microcapsules prepared from blends of chitosan with starch and/or pullulan had lower aw values than those prepared from chitosan alone. Oxidative stability was evaluated by measuring oxidation induction time (OIT) using pressure differential scanning calorimetry. OIT values ranged from 14 to 20 minutes. Microcapsules prepared from chitosan with lower DD and higher MW had longer OITs than those prepared from chitosan with higher DD and lower MW. Microcapsules prepared from blends of chitosan, starch, and pullulan had longer OITs than those prepared from chitosan alone. Oxidative stability of microcapsules during long term storage was evaluated on one microcapsule formulation by monitoring peroxide value (PV) and secondary oxidation products by HS-SPMEGC/ MS. Volatiles including propanal, 1-penten-3-ol, pentanal, hexanal, and 2,4-heptadienal were detected in the headspace of the microcapsules; however, PVs did not indicate substantial oxidation of the ω3-PUFA source during 5 weeks of storage. Chitosan, high-amylose starch, and pullulan are effective materials for microencapsulation of ω3 PUFA sources. / Ph. D.

This research first examined nitrification inhibition caused by different classes of industrially relevant chemicals on activated sludge and found that conventional aerobic nitrification was inhibited by single pulse inputs of every chemical tested, with 1-chloro-2,4-dinitrobenzene (oxidant) having the most severe impact, followed by alkaline pH 11, cadmium (heavy metal), cyanide, octanol (hydrophobic) and 2,4-dinitrophenol (respiratory uncoupler). Of the different chemicals tested, the oxidative and hydrophobic chemicals showed severe nitrification inhibition relative to other treatment processes and therefore deserved further investigation. For oxidative chemicals, we hypothesized that the more severe inhibition was because nitrifying bacteria lack one or more of the microbial stress response mechanisms used to mediate the toxic effect of oxidative chemicals. During these experiments, we showed that a rapid (minutes) antioxidant potassium efflux mechanism does not exist in two nitrifying bacteria, Nitrosomonas europaea and Nitrospira moscoviensis. Furthermore, we showed that another important antioxidant molecule, glutathione, was not oxidized as readily as in a non-nitrifying bacterium. Furthermore, we hypothesized that hydrophobic chemical-induced nitrification inhibition recovered more quickly because of the presence of membrane modification stress response mechanisms. While testing this hypothesis, we showed that N. europaea modified its cell membrane in response to hydrophobic chemicals using a long-term (hours) membrane modification mechanism that required the synthesis of new fatty acids, but it did not contain a short-term (minutes) response mechanism involving a cis/trans isomerase. Therefore, investigating these nitrifier stress responses showed that nitrifiers lack short-term stress responses that may be used to rapidly detect inhibition, indicating that conventional methods of detecting nitrification inhibition, like differential respirometry and nitrate generation rate (NGR), are still the fastest and easiest methods to use. Because several conventional methods exist, we also investigated differences between differential respirometry and a UV method we developed to measure NGR. During these tests, we showed that the UV NGR method provided a more reliable measure of nitrification inhibition than differential respirometry, and that the time to maximum nitrification inhibition depended on the properties of the chemical toxin, which implies that longer exposure times may be needed to accurately predict nitrification inhibition. / Ph. D.