Thesis (M.S.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed Nov. 11, 2008). Includes bibliographical references.

Le bruissement enzymatiques, appelé mélanose post mortem chez les crustacés est un phénomène enzymatique catalysé par des protéines à activités phénoloxydases (tyronase, catécholase, laccase et hémocyanine). L'utilisation de conservateurs de type sulfites (E220 à E228 et E539) reste à l'heure actuelle la solution la plus répandue pour éviter le développement de cette coloration peu attrayante pour le consommateur. Mais une partie de la population développe des réactions d'hypersensibilité en consommant des aliments sulfités. Dans l'onbjectif de rechercher une alternative à ces conservateurs, deux axes de recherche ont été développés durant ces travaux de thèse : la caractérisation biochimique des protéines responsables de la mélanose post mortem chez la crevette P. monodon, puis la recherche de molécules inhibitrices. Un fractionnement sur résine Phenyl Sepahrose™ CL-4B (HIC) suivie d'une séparation par électrophorèse SDS-PAGE ont montré la présence de trois protéines de 46, 82 et 89 kDa à activité principalement laccase. Une identification par RP-HPLC-Q/TOF a mis en évidence la présence d'hémocyanine uniquement. Un pH de 7,0 et une température comprise entre 37 et 50 °C ont mis en évidence les activités les plus importantes, en utilisant le dosage enzymatique dit "test au MBTH". Par ailleurs, un criblage à haut débit de 45 molécules potentiellement inhibitrices a été réalisé dans des conditions d'analyses standardisées grâce à l'outil de robotique de la plateforme Realcat. Une inhibition a été mise en évidence pour 23 composés, certains étant suffisamment efficaces pour être utilisés seuls. D'autres pourraient être introduits dans un cocktail de molécules inhibitrices aux fonctionnalités complémentaires. Les résultats des tests de trempage réalisés sur des crevettes entières ont montré qu'il était indispensable de compléter les études in vitro avec des essais à l'échelle de la matrice alimentaire dans son intégralité. / Enzymatic browning, called post mortem melanosis in crustaceans, is an enzymatic phenomenon catalyzed by proteins with phenoloxidase activities (tyronase, catecholase, laccase and hemocyanin). The use of sulfite preservatives (E220 to E228 and E539) remains at present the most widespread solution to avoid the development of this unattractive color towards consumers. But, a part of the population develops hypersensitivity reactions by consuming sulfited foods. With the objective to find an alternative to these conversators, two research axes have been planned : the biochemical characterization of the proteins responsible for post mortem melanosis in the P. monodon shrimp, then the search for inhibitory molecules. Fractionation on Phenyl Sepahrose™ CL-4B resin (HIC) followed by SDS-PAGE electrophoresis separation showed the presence of three proteins of 46, 82 and 89 kDa with mainly laccase activity. Identification by RP-HPLC-Q / TOF revealed the presence of hemocyanin only. A pH of 7.0 and a temperature between 37 and 50 °C showed the most important activities, using the enzymatic assay called "MBTH test". On the other hand, a high throughput screening of 45 potentially inhibitory molecules could be performed under standardized analysis conditions thanks to the robotic tools of the Realcat platform

Call number: LD2668 .T4 1986 M375 / Master of Science / Chemistry

Hypoxia and sulfide exposure, increased using glucose, are considered major environmental stressors in seagrass communities. Quantum efficiency, total soluble protein and catalase activity were quantified to evaluate the applicability of each of these bioindicators to detect environmental stress in three tropical seagrass species, Thalassia testudinum (Banks ex Kèoenig), Halodule wrightii (Ascherson) and Syringodium filiforme (Kuetz). Hypoxia + sulfide treatments significantly decreased the quantum efficiency of all three species, but showed no response in protein and catalase activity. Although no treatment effect was found, catalase activity was enhanced in T. testudinum leaves and H. wrightii roots relative to other tissues, while S. filiforme showed no location-specific catalase activity. These results indicate that quantum efficiency is a more sensitive indicator than protein and catalase activity to hypoxia and sulfide stress in seagrasses. / by Connor Irwin. / Title on signature page (p. ii) : Environmental stress biomonitoring in sub-tropical seagrasses. / Thesis (M.S.)--Florida Atlantic University, 2010. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2010. Mode of access: World Wide Web.

Pharmaceutical compounds represent a rapidly emerging class of environmental contaminants. Such compounds were recently classified by the U.S. Geological Survey, including several antibiotics. An LC-MS/MS screening method for the top five antibiotics in drinking water was developed and validated using a Shimadzu LC-MS-IT-TOF. The separation was performed using a Waters Acquity UPLC BEH C18 column with a gradient elution. Sulfamethazine was exposed to conditions intended to mimic drinking water chlorination, and samples were collected and quenched with excess sodium sulfite. Kinetics of sulfamethazine degradation was followed as well as the formation of the major chlorinated byproduct (m/z 313). For the screening method, all five antibiotic peaks were baseline resolved within 5 minutes. Additionally, precision and accuracy of the screening method were less than 15%. Degradation of sulfamethazine upon exposure to drinking water chlorination occurred by first order kinetics with a half-life of 5.3 × 10(4) min (approximately 37 days) with measurements starting 5 minutes after chlorination. Likewise, the formation of the major chlorinated product occurred by first order kinetics with a rate constant of 2.0 × 10(-2). The proposed identification of the chlorinated product was 4-amino-(5-chloro-4,6-dimethyl-2-pyrimidinyl)-benzenesulfonamide (C12H13N4O2SCl) using MS (n) spectra and databases searches of SciFinder and ChemSpider.

Brettanomyces bruxellensis est un microorganisme qui est considéré comme la cause majeure des défauts microbiologiques du vin. L’importance de cette levure à l’échelle industrielle est liée au fait qu’elle est isolée à partir de substrats différents tels que la bière, le kombucha, les molasses utilisées pour la production de bioéthanol et autres. Ce projet a pour objectif d’étudier la diversité génétique de l’espèce en se basant sur une large population d’isolats provenant de niches écologiques et géographiques variées. Pour ce faire, une méthode de génotypage robuste (analyse microsatellite) a été optimisée et appliquée sur la population, mettant en évidence la coexistence de populations diploïdes et triploïdes à l’échelle globale. Puis, la relation entre regroupement génétique et traits physiologiques a été explorée. Notamment, l'étude de la tolérance aux sulfites a été effectuée sur un sous-ensemble de souches représentatif de la population. Les résultats obtenus mettent en évidence un lien entre groupes génétiques et comportement vis-à-vis des sulfites. Des expériences de compétition en présence de dioxyde de soufre montrent un avantage sélectif des souches tolérantes aux sulfites par rapport aux souches sensibles, suggérant ainsi une adaptation spécifique au principal antiseptique utilisé en œnologie. Ce travail contribue à une meilleure connaissance de cette levure d’altération du vin en termes de diversité génétique et phénotypique et permet d’émettre des hypothèses sur les stratégies évolutives d'adaptation au milieu anthropique de cette espèce modèle non conventionnelle. / Brettanomyces bruxellensis is a microorganism described as the first cause of microbial spoilage of wine. Its industrial relevance is highlighted by the fact that this yeast is isolated from different substrates such as beer, kombucha, bioethanol fermentation molasses and others. This project aims to explore the genetic diversity of the species by studying a large population of isolates from various geographical and ecological niches. For this purpose, a robust genotyping method (microsatellite analysis) was optimized and applied on the population, thus highlighting the coexistence of diploid and triploid populations worldwide. Further, the relation between genotypic clustering and physiological traits was studied. Namely, sulphite tolerance assay was performed on a subset of strains representative of the total population. The results reveal a link between genetic group and growth profile in the presence of sulphur dioxide. Competition experiments in presence of sulphites highlight a selective advantage of sulphite tolerant strains compared to sulphite sensitive ones, thus suggesting a specific adaptation to the main antimicrobial used in winemaking. This work contributes to a deeper understanding of this wine spoilage microorganism in means of genetic and phenotypic diversity and sheds light on putative evolutionary strategies for adaptation to human related environment of this non-conventional model yeast species.

Ces dernières années, de plus en plus de composés organiques réfractaires et toxiques ont été détectés dans les eaux usées. Un bon nombre de ces polluants organiques sont difficilement dégradés par des traitements classiques. Les procédés d’oxydation avancée à base de radicaux sulfates (SR-AOP) sont apparus comme une méthode innovante dans le domaine de la décontamination oxydative des eaux polluées. Des études antérieures ont porté sur ces SR-AOP utilisant du peroxodisulfate (PS) ou du peroxomonosulfate (PMS) comme oxydants, en particulier des couples «métaux de transition et oxydants» (systèmes Fe (II)/PS, systèmes Ni(II)/PMS et Co (II))/PMS), où il a été confirmé que SO4•-·présentent des avantages (sélectivité) par rapport au radical hydroxyle (HO•) pour la décontamination des eaux usées contenant des polluants organiques.Dans cette thèse, nous avons généré des radicaux tels que le radical sulfite SO3•-, le radical sulfate SO4•-, le radical peroxomonosulfate SO5•- à partir d’ions métalliques (Cr(VI), le Co(II), le Fe(III)) capables d’activer le sulfite pour la dégradation des composés organiques. L'efficacité d'élimination et le mécanisme d'oxydation ont été étudiés et le rôle des espèces soufrées a été élucidé. / In recent years, more and more refractory and toxic organic compounds are detected in wastewater. Many of these organic pollutants can hardly be degraded by conventional water treatments. Sulfate radical based advanced oxidation process (SR-AOPs), have emerged as a promising method in the field of oxidative decontamination of polluted water. Past studies focused on this SR-AOPs with peroxydisulfate (PS) or peroxymonosulfate (PMS) as oxidants, especially the ‘transition metal + oxidants’ (i.e. Fe(II)/PS system, Ni(II)/PMS system and Co(II)/PMS system), which has been confirmed that SO4·− has advantages over HO in the decontamination of wastewater containing organic pollutants. In this PhD thesis, oxysulfur radicals including sulfite radical SO3·−, sulfate radical SO4·−, peroxymonosulfate radical SO5·− produced by transition metal ions such as Cr(VI), Co(II), Fe(III) activated sulfite were used to degrade organic compounds. The removal efficiency, the oxidation mechanism were examined, and and the role of sulfur species were elucidated.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / The food industry uses chemical additives to extend the shelf life of its products and maintain their sensory and microbiological characteristics acceptable. However, the presence of preservatives in foods may interfere in the quantification of total antioxidant activity. Therefore, this study aimed to evaluate the influence of food preservatives on total antioxidant activity and on the sensory and microbiological quality of acerola tropical juice. The fruits were washed, sanitized (200 ppm sodium hypochlorite), pulped and then were formulated with water and its specific preservatives. The juices were heat treated (90ÂC/60 seconds) and hot filled in glass bottles, which were closed, cooled until 35ÂC and stored at 25ÂC for 180 days. The formulations tested were: control juice without additive (C); juice with addition of 0.004 g/100 mL of sodium metabisulphite (SMS); juice with addition of 0.08 g/100 mL of potassium sorbate (SSP); juice added of 0.05 g/100 mL of sodium benzoate (SBS); juice added of 0.002 g/100 mL of sodium metabisulfite and 0.04 g/100 mL of potassium sorbate S(MS+ SP); juice with addition of 0.002 g/100 mL of sodium metabisulfite and 0.025 g/100 mL of sodium benzoate S(MS + BS). It was carried out chemical determinations for ascorbic acid, total carotenoids, total anthocyanins, total extractable polyphenolics and total antioxidant activity by ABTS and DPPH methods. Commercial sterility testing was performed on samples to determine the hygienic and sanitary processing and to check for any microbiological changes. In sensory acceptance tests were used acerola nectars, prepared by dilution from the acerola tropical juice and added sucrose to obtain 11 ÂBrix, using a 9-point hedonic scale for flavor, color, aroma, sweetness, appearance, body and overall impression. The experiments were performed at three time points (0, 90 and 180 days) and the determinations were made in triplicate. The results were analyzed by the interaction between the juice type and storage time. Regarding anthocyanins content, all samples showed an average of 1.0 mg/100mL (P> 0.05). The acid ascorbic contents remained stable during the storage and ranged from 256.22 mg/100mL (C juice) to 301mg/100mL (SMS and SSP juices). The total carotenoid in treatments with or without preservatives did not vary significantly among themselves, with mean values of 0.04 mg/100mL. The yellow flavonoids showed no significant difference between the juice type (P> 0.05), averaging 5.30 mg/100 mL. For soluble dark pigments, SMS obtained the lowest range in absorbance at 90 days time (0.170 to 0.240), and greater variation for the control juice (from 0.230 to 0.320). All samples showed high losses of phenolics during storage, observing greater losses at times of 0 and 90 days, which ranged from 36.59% for S(BS + MS) and 42.97% for SMS. There was a significant variation of total antioxidant activity with time by the DPPH test (P≤0.05). The values ranged from 877.41 to 978.44 g/g DPPH for C and SMS juices, respectively. A similar behavior was observed for contents analyzed by the ABTS testing: 58.46 mM Trolox/g for SMS and 49.80 mM Trolox/g for control juice. A significant and positive correlation was verified between polyphenols and ABTS (r = 0.78) and DPPH (r = 0.44); anthocyanins and ABTS (r = 0.39) and DPPH (r = 0.44). Yellow flavonoids and ABTS and DPPH assays were negatively and significantly correlated (r = -0.83 and r = -0.52), respectively. There was no significant correlation between ascorbic acid and ABTS and DPPH assays. There was no significant interaction between any treatment and storage time (P> .05). Sodium metabisulphite and its combinations did not influence sensorial acceptance of acerola nectars, with assessments within the acceptable range for all analyzed attributes along storage time. The potassium sorbate and sodium benzoate singly were the additives that contributed most to a higher rejection sensory of products. The employed chemical preservatives, associated with the heat treatment, were effective for maintaining the commercial sterility of acerola tropical juices. / A indÃstria de alimentos utiliza-se de conservantes quÃmicos para prolongar a vida Ãtil de seus produtos e manter suas caracterÃsticas sensoriais e microbiolÃgicas aceitÃveis. Contudo a presenÃa desses conservantes nos alimentos pode interferir na quantificaÃÃo da atividade antioxidante total. Portanto, este trabalho objetivou avaliar a influÃncia dos conservantes alimentares na atividade antioxidante total, na qualidade sensorial e microbiolÃgica de sucos tropicais de acerola. As frutas foram lavadas, sanitizadas (200mg/L de cloro ativo) e despolpadas, em seguida, formuladas com Ãgua e seus conservantes especÃficos. Os sucos foram tratados termicamente (90ÂC/60 segundos) e acondicionados a quente em garrafas de vidro, que foram fechadas, resfriadas atà 35ÂC e armazenadas à temperatura de 25ÂC por 180 dias. As formulaÃÃes testadas foram: suco controle, sem aditivo (C); suco com adiÃÃo de 0,004 g de metabissulfito de sÃdio/100 mL (SMS); suco com adiÃÃo de 0,08g sorbato de potÃssio /100 mL de (SSP); suco adicionado de 0,05g de benzoato de sÃdio /100 mL (SBS); suco com 0,002g de metabissulfito de sÃdio e 0,04 g/100 mL de sorbato de potÃssio/100 mL S(MS+SP)); suco com adiÃÃo de 0,002g/100 mL de metabissulfito de sÃdio e 0,025 g/100 mL de benzoato de sÃdio S(MS+BS). Foram realizadas determinaÃÃes quÃmicas de Ãcido ascÃrbico, carotenÃides totais, antocianinas totais, polifenÃis extraÃveis totais e quantificaÃÃo da atividade antioxidante total pelos mÃtodos ABTS e DPPH. Foi realizado o teste de esterilidade comercial nas amostras para determinar as condiÃÃes higiÃnico-sanitÃrias do processamento e verificar as possÃveis alteraÃÃes microbiolÃgicas. Os testes sensoriais de aceitaÃÃo foram utilizados nÃctares de acerola, preparados por diluiÃÃo a partir do suco tropical de acerola e adicionado de sacarose atà obtenÃÃo de 11 ÂBrix, os quais foram avaliados quanto aos atributos impressÃo global, cor, aparÃncia, aroma, doÃura, sabor e corpo utilizando uma escala hedÃnica estruturada de nove pontos. Os experimentos foram realizados em trÃs tempos (0, 90 e 180 dias) e realizados em triplicata. Os resultados foram tratados por anÃlise de interaÃÃo entre os diferentes tratamentos e tempo de armazenamento. Para o teor de antocianinas, todas as amostras apresentaram mÃdia de 1,0mg/100g (P>0,05). O teor de Ãcido ascÃrbico se manteve estÃvel durante o armazenamento, variando de 256,22 mg/100mL para o suco C, a 301mg/100mL, para os sucos SMS e SSP. Os teores de carotenÃides totais nos tratamentos com ou sem conservantes nÃo diferiram entre si (P>0,05), apresentando valor mÃdio de 0,04mg/100mL. Os flavonoides amarelos nÃo apresentaram diferenÃa significativa (P>0,05), apresentando mÃdia de 5,30mg/100mL. Para pigmentos escuros solÃveis o SMS obteve a menor variaÃÃo na absorbÃncia no tempo de 90 dias (0,170 a 0,240), e a maior variaÃÃo para o suco controle (0,230 a 0,320). Todas as amostras apresentaram perdas elevadas de fenÃlicos durante o armazenamento, observando-se perdas maiores entre 0 e 90 dias, que variaram de 36,59% para S(MS+BS) e 42,97% para SMS. Houve variaÃÃo significativa com o tempo para a atividade antioxidante total pelo ensaio DPPH (P≤0,05). Os valores variaram entre 877,41 e 978,44 g/g DPPH para os sucos C e SMS, respectivamente. Comportamento semelhante foi observado para os teores analisados pelo mÃtodo ABTS, observando-se para o SMS, 58,46 ÂM Trolox/g; e para o suco controle, 49,80 ÂM Trolox/g. A correlaÃÃo positiva e significativa foi verificada entre polifenÃis e o ensaio ABTS (r = 0,78) e para DPPH (r = 0,44). Antocianinas totais e ABTS (r = 0,39) e DPPH (r = 0,44). Os flavonÃides amarelos e os ensaios ABTS e DPPH se correlacionaram negativamente e significativamente (r = -0,83 e r= -0,52; respectivamente). NÃo foi observada interaÃÃo significativa entre os tratamentos e os tempos de armazenamento (P > 0,05). O metabissulfito de sÃdio e suas combinaÃÃes nÃo influenciaram na aceitaÃÃo sensorial dos nÃctares de acerola, com avaliaÃÃes dentro da faixa de aceitaÃÃo para todos os atributos analisados ao longo do tempo de armazenamento. Os aditivos sorbato de potÃssio e benzoato de sÃdio isoladamente contribuÃram para uma maior rejeiÃÃo sensorial dos produtos. Os conservantes quÃmicos utilizados, associados ao tratamento tÃrmico empregado, foram eficientes para a manutenÃÃo da esterilidade comercial dos sucos tropicais de acerola.