Thesis (MScAgric)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: In recent years South African wines have been under the spotlight due to references in the international wine media to a distinctive ‘burnt rubber’ character. Many winemakers and wine experts argued that the peculiar character could be ascribed to winemaking errors linked to mismanaged fermentation. An alternative possible source of the taint was identified in the coal tar creosote used as a wood preservative in vineyard trellis systems. South African regulations allow for the use of creosoted utility poles in agricultural land, but in Europe and USA their usage has been highly regulated and even banned for certain applications. Contamination of foodstuff by polycyclic aromatic hydrocarbons (PAHs) is one of the major motives for the banning of creosote in Europe and USA. Some of the compounds in the creosote mixture display very strong odour characteristics and for this reason it became the focus of attention for the present study. The overall aim of this study was to determine if vines trellised with creosoted posts could accumulate or absorb the various malodorous compounds deriving from the wood treatment onto the grape berries. These compounds could then be extracted from the grape berries into the wine during alcoholic fermentation, creating quality and sensory problems. Chapter 2 of this thesis gives an overview of the extensive literature that deals with methods of analysis of PAHs and malodorous phenols using both Gas Chromatography (GC), as well as High Performance Liquid Chromatography (HPLC). New methods for sample preparation, as well as analysis of PAHs using HPLC-Diode Array Detector (DAD), were developed and the results reported in Chapter 3. It was demonstrated that Sauvignon blanc experimental wines contained only chrysene at very low levels. The concentrations of chrysene found in the experimental wines are within the prescribed parameters as established by The Commission of the European Communities. Since no other PAH compounds were found in the samples analysed, it was concluded that the experimental wines were safe for human consumption and complied with EU regulations. The effect of vineyard posts on the sensorial characteristics of wine is discussed in Chapter 4. Creosoted poles were found to be responsible for an off-flavour described as ‘burnt rubber’ and ‘tarry’ in Merlot wines produced from grapes grown in close proximity to the posts. Following some of the reported findings, new guidelines have been introduced in the Integrated Production of Wine certification, which advise against the use of creosoted poles for vineyard trellising. This preliminary but important guideline will bring the South African wine industry a step closer to the fulfilling the obligations for food safety as required by the legislation of our major export partners. Future investigations are recommended to completely understand and evaluate the cumulative effect of creosoted posts in a fully trellised vineyard. / AFRIKAANSE OPSOMMING: Oor die afgelope paar jaar is Suid-Afrikaanse wyne onder die soeklig geplaas as gevolg van verwysings in die internasionale wynmedia na ‘n duidelike ‘gebrande rubber’-karakter. Baie wynmakers en wyndeskundiges het aangedui dat hierdie besonderse karakter toegeskryf kan word aan wynbereidingsfoute wat verband hou met gisting wat wanbestuur is. ‘n Alternatiewe moontlike oorsprong van die smaak is geïdentifiseer in die koolteer wat as ‘n houtpreserveermiddel in wingerdopleistelsels gebruik word. Suid-Afrikaanse regulasies maak voorsiening vir die gebruik van kreosoteerde nutspale in landbougrond, hoewel hulle gebruik in Europa en die VSA hoogs gereguleerd en in sommige gevalle selfs verbied is. Die besmetting van kossoorte deur polisikliese aromatiese koolwaterstowwe (polycyclic aromatic hydrocarbons (PAHs)) is een van die vernaamste redes vir die verbanning van kreosoot in Europa en die VSA. Sommige van die verbindings in die kreosootmengsel het baie sterk geurkenmerke en daarom is dit die fokus van die huidige studie. Die oorhoofse doelwit van hierdie studie was om te bepaal of wingerde wat op kreosoteerde pale opgelei is, die verskillende onwelriekende verbindings afkomstig van die houtbehandeling in die druiwekorrels kan akkumuleer of absorbeer. Hierdie verbindings sou dan tydens alkoholiese gisting uit die druiwekorrels in die wyn geëkstraheer kon word, wat aanleiding sou gee tot kwaliteits- en sensoriese probleme. Hoofstuk 2 van hierdie tesis verskaf ‘n oorsig van die breedvoerige literatuur wat handel oor metodes om PAH’s en onwelriekende fenole met behulp van beide gaschromatografie (GC) en hoëdrukvloeistofchromatografie (HPLC) te analiseer. Nuwe metodes is ontwikkel om monsters voor te berei en om PAH’s met behulp van ‘n HPLC-diode array detector (DAD) te analiseer. Die resultate word in Hoofstuk 3 gerapporteer. Daar is aangetoon dat die eksperimentele Sauvignon blanc-wyne chriseen teen baie lae vlakke bevat het. Die konsentrasies van chriseen wat in die eksperimentele wyne gevind is, is binne die voorgeskrewe parameters van die Kommissie van die Europese Gemeenskap. Aangesien daar nie ander PAHverbindings in die geanaliseerde monsters gevind kon word nie, is daar tot die gevolgtrekking gekom dat die eksperimentele wyne veilig is vir menslike verbruik en aan die EG-regulasies voldoen. Die effek van wingerdpale op die sensoriese kenmerke van wyn word in Hoofstuk 4 bespreek. Kreosoteerde pale is gevind wat verantwoordelik is vir ‘n wangeur in Merlot-wyne afkomstig van druiwe wat naby die pale gegroei het en wat as ‘gebrande rubber’ en ‘teeragtig’ beskryf is. Op grond van sommige van die gerapporteerde bevindings, is nuwe riglyne ingesluit in die sertifisering vir die Geïntegreerde Produksie van Wyn, wat aanbeveel dat kreosoteerde pale nie vir wingerdoplei gebruik word nie. Hierdie voorlopige, maar belangrike riglyn sal die Suid-Afrikaanse wynbedryf al beter in staat stel om te voldoen aan die voedselveiligheid regulasies wat vereis word deur die wetgewing van ons belangrikste uitvoervennote. Toekomstige ondersoeke moet aangewend word om die kumulatiewe effek van kreosoteerde pale in volledig opgeleide wingerde ten volle te verstaan en te evalueer. / Indaba Scholarship and Winetech for their financial support

Doctor of Philosophy / Department of Chemistry / Duy H. Hua / Our research work is divided into three chapters. In the first chapter, synthesis of substituted phenolic compounds including halogenated di- and trihydroxybenzenes, aminophenols, and substituted di-tert-butylphenols, their redox potential, laccase oxidation, and mosquito anti-larval activities are discussed. The synthesized substituted phenols were found to be the substrates but not the inhibitors of laccase. An inverse correlation between the oxidation potential and the laccase oxidation efficiency of halogenated hydroxybenzenes and aminophenols was established. However, substituted di-tert-butylphenols were found to have anti-larval activities in mosquitoes resulting in the death of the larvae just before reaching pupation. Among the di-tert-butyl phenols studied, water insoluble, 2,4-di-tert-butyl-6-(3-methyl-2-butenyl)phenol (16), 2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-methylpropanal oxime (14), and 6,8-di-tert-butyl-2,2-dimethyl-3,4-dihydro-2H-chromene (17) caused the mortility of 98%, 93%, and 92% of Anopheles gambiae larvae in the concentration of 182 nM, 3.4 µM, and 3.7 µM, respectively. In particular, compound 16 had similar anti-larval activities as compared to MON-0585, an anti-larval agent reported by Monsanto in the 70’s. In the second chapter, inhibition of protein kinase C (PKC) phosphorylation by substituted quinolines (PQs) is inverstigated. PQ compounds such as N-(3-aminopropyl)-6-methoxy-4-methyl-5-(3-(trifluormethyl)phenoxy)quinolin-8-amine (PQ1), N-(furan-2-ylmethyl)-6-methoxy-4-methyl)-5-(3-(trifluoromethyl)phenoxy)quinolin-8-amine (PQ11), and 6-methoxy-4-methyl-N-(quinolin-4-ylmethyl)-5-(3-(trifluoromethyl)phenoxy)quinolin-8-amine (PQ15) were found to inhibit PKC phosphorylation with IC50 values of 35 nM, 42.3 nM, and 216.3 nM respectively, among which PQ1 and PQ11 were found to be potent PKC inhibitors as comparable to that of staurosporine (IC50 = 33 nM). In chapter three, the tissue distribution of PQ1 and PQ11 in normal C57BL/6J mice and the effect of PQ1 on the normal tissues of mice were investigated. Substituted quinolines, PQ1 and PQ11 were distributed in the tissues in concentrations that were more than 40 folds of their effective dose. PQ1 and PQ11 were also found to penetrate the blood brain barrier and collect in the tissue in significant amounts. The administration of PQ1 and PQ11 had no effect in the normal behavior of the animals indicating no short term adverse effects. PQ1 was found to increase the expression of survivin, an anti-apoptotic factor and decrease the expression of cleaved caspase-3 and caspase-8, pro-apoptotic proteins. These studies suggests that PQ1 might have anti-apoptotic activities in normal cells, in contrast to the role of PQ1 in cancer cells where it has demonstrated to induce apoptosis. The study also indicated that PQ11 was better metabolized from the tissues over time as compared to PQ1.

Doctor of Philosophy / Department of Chemistry / Duy H. Hua / Three research projects were carried out and they are described below. The synthesis of substituted phenolic compounds including halogenated di- and trihydroxybenzenes, aminophenols, and substituted di-tert-butylphenols are described. Redox potentials of the synthesized molecules along with various known laccase substrates were measured, and an inverse relationship between the oxidation potential and the efficiency of oxidation by laccase of halogenated hydroxybenzenes and aminophenols is demonstrated. The synthesized substituted phenols were found to be substrates but not inhibitors of laccase. We discovered a new class of di-tert-butylphenols compounds that inhibits the growth of mosquito larvae at low concentrations. Compound 17, 2,4-di-tert-butyl-6-(3-methyl-2-butenyl) phenol caused greater than 98% mortality of third-instar larvae of Anopheles gambiae in the concentration of 0.18 µM. These compounds do not inhibit laccases. It appears that they affect a new target of the mosquito that is different from those of currently existing pesticides. Two anti-Alzheimer molecules, CP2 and TP70, discovered in our laboratory were studied for their pharmacokinetics and distribution. The distribution of CP2 and TP70 in mouse brain region and various tissues of mice were examined. HPLC analysis revealed that CP2 treatment in primary neurons accumulates in mitochondria fraction. Similarly, the amount of CP2 in the brain tissue from wild type and APP/PS1 mice treated with 25 mg/kg/daily for 2 months also have the highest concentration in the mitochondria fractions in the hippocampus. The results show that CP2 and TP70 can penetrate the blood brain barrier and accumulate in the tissue in significant amounts. Pharmacokinetics and bioavailability of compound TP70 were determined. Area under the curve and bioavailability value F were calculated, and data show that TP70 has a good PK profile and bioavailability. For the preparation of a novel tripeptidyl norovirus 3C-like protease (3CL[superscript]pro) inhibitor, the P3 unnatural amino acid, (S)-3-hydroxyphenylalanine was synthesized. The P3 is designed to increase the polarity with the addition of the alcohol group. After combining the P3 unnatural amino acid with the P1 and P2 to form the novel tripeptidyl compound, a study comparing the relations between the structure and its activity (SAR) will confirm whether prediction is correct in our pursuit for an antiviral therapeutic drug in the form of a protease inhibitor.

Óleos contendo alta proporção de ácidos graxos ômega-3 (n-3 FA) têm sido aplicados na formulação de alimentos ou comercializados como suplementos, com a finalidade de reduzir o risco cardiovascular, principalmente devido aos seus efeitos hipotriglicêmicos e anti-inflamatórios. No entanto, a susceptibilidade à oxidação dos n-3 FA é elevada, levando à formação de vários produtos secundários, incluindo alguns tóxicos e potencialmente aterogênicos. Por esta razão, compostos naturais com propriedades antioxidantes têm sido investigados com o objetivo de melhorar a estabilidade oxidativa dos óleos com alta proporção de n-3 FA. Neste estudo, avaliou-se a capacidade antioxidante de dois compostos naturais (ácido sinápico e hidrato de rutina) utilizando-se um modelo acelerado para oxidar os óleos. Foram combinados cinco indutores (Temperatura; Ferro- Fe2+; 2,2\'-Azobis dicloridrato de 2-amidinopropano - AAPH; ascorbil palmitato - AP e 2,2\'-azobis -2,4-dimetilvaleronitrilo - AMVN) em um delineamento fatorial (25-1) com ½ fração de \"resolução V\" para acelerar a oxidação de três óleos (linhaça, Echium e peixe) contendo diferentes fontes de n-3 FA: ácido α-linolênico (ALA), ácido estearidônico (SDA) e ácido eicosapentaenóico (EPA) + ácido docosahexaenóico (DHA), respectivamente. Não houve diferença entre os marcadores de oxidação (LOOH e TBARS) estimados pelos modelos e os valores observados experimentalmente. Os indutores AMVN e Fe2+ foram os principais fatores responsáveis pelo aumento da concentração de TBARS. Os valores dos marcadores oxidativos obtidos 48 h após a indução foram semelhantes ou superiores àqueles observados nas amostras oxidadas a 60°C por 15 dias, sendo ambos maiores que os valores observados nas amostras de óleo frescas. Entre os compostos voláteis formados pela oxidação de diferentes fontes de n-3 FA, (E, E) 2,4 -heptadienal, (E, E) 2,4-decadienal, decanal, undecanal e (E) -2-undecenal foram identificados em todas as amostras, podendo ser utilizados como marcadores oxidativos mais específicos. Utilizando o modelo de oxidação acelerada, o hidrato de rutina melhorou a estabilidade oxidativa do óleo de peixe, provavelmente devido à presença de grupos catecol em sua estrutura química. Este estudo contribuiu para que ensaios mais rápidos fossem realizados na avaliação do efeito antioxidante de novas moléculas aplicadas em óleos funcionais comestíveis. / Oils containing a high proportion of omega-3 fatty acids (n-3 FA) have been used in the formulation of foods or sold as supplements, aiming to reduce cardio-vascular risks, mainly due to their hypotriglycemic and anti-inflammatory effects. However, n-3 FA are highey susceptible to oxidation, leading to the formation of several products, including some toxic and potentially atherogenic. For this reason, natural products with antioxidant properties have been investigated to improve the oxidative stability of oils with a high proportion of n-3 FA. This study evaluated the antioxidant capacity of two natural compounds (sinapic acid and rutin hydrate), using an accelerated model to oxidize the oils. Five inducers were combined (Temperature, Iron-Fe2+, 2,2\'-Azobis (2-amidinopropane) dihydrochloride-AAPH, Ascorbyl palmitate-AP and 2,2\'-azobis-2,4-dimethylvaleronitrile-AMVN) in a factorial design (25-1) ½ fraction of \"resolution V\" to accelerate the oxidation of three oils (flaxseed, Echium and fish) containing different sources of n-3 FA: α-linolenic acid (ALA), stearidonic acid (SDA) and eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA), respectively. There was no difference between the oxidation markers (LOOH and TBARS) estimated by the regression models and the values experimentally observed. The inducers AMVN and Fe2+ were the main factors responsible for the increase of TBARS concentration. The values of the oxidation markers obtained 48h after the induction were similar to or higher than those obtained when the samples were oxidized at 60°C for 15 days, both being more elevated than the values observed in the fresh oils. Among the volatile compounds formed by the oxidation of different sources of n-3 FA, (E, E) 2,4-heptadienal, (E, E) 2,4-decadienal, decanal, undecanal and (E)-2-undecenal were identified in all samples, and could be used as more specific oxidation markers. Using the accelerated model, rutin hydrate improved the oxidative stability of fish oil, probably due to the presence of catechol groups in its chemical structure. This study showed that faster anays could be performed to evaluate the antioxidant effect of new molecules applied on edible functional oils.

Com o intuito de viabilizar, ambiental e economicamente, a mineralização de substâncias fenólicas presentes em águas e efluentes, foram estudadas as moléculas do timol (C10H14O TOH, a 200 mg L-1) e do bisfenol-A (C15H16O2 BPA, a 25 mg L-1) utilizando Processos Oxidativos Avançados (POA), no caso, ozônio (O3), radiação ultravioleta (UV) e peróxido de hidrogênio (H2O2) que, quando combinados, ou em condições específicas, promovem a geração de radicais hidroxila (OH), um poderoso oxidante. O sistema de tratamento utilizado foi desenvolvido no próprio Laboratório de Tratamento de Resíduos do CENA/USP. O TOH e o BPA foram submetidos a tratamento em pH 3 e 11, com O3 (1,98 g h-1) e UV (0, 8 e 32 watts). Adicionalmente, o TOH foi tratado também com H2O2 (0, 1.176 e 2.352 mg), totalizando 18 tratamentos para o TOH e 6 para o BPA. A eficiência dos tratamentos foi avaliada a partir da porcentagem de remoção do carbono orgânico total (TOC). Para ambas as moléculas, a combinação 1,98 g h-1 de O3, 32 watts UV e ausência de H2O2, em pH 3, apresentou os melhores resultados de conversão de TOC a CO2, especificamente 99,6 ± 0,6% em 90 minutos para o timol e 93,9 ± 0,6% de mineralização de bisfenol-A em 60 minutos de tratamento. Este tratamento foi então reproduzido no resíduo de timol gerado pelos laboratórios do CENA/USP e o resultado obtido foi de 92,64 ± 1,2%, um pouco inferior ao obtido para a solução de TOH, provavelmente devido à presença de outras substâncias. As soluções (TOH e BPA) e o resíduo (TOH) tratados foram submetidos à análise de fenóis totais, obtendo-se resultados inferiores ao exigido (0,5 mg L-1 de fenóis totais) pelas resoluções CONAMA 357/2005 e 397/2008 para a disposição em corpos hídricos. Realizaram-se também ensaios de toxicidade com os organismos Pseudokirchneriella subcapitata, Daphnia magna e Hydra attenuata (que representam os distintos níveis tróficos da cadeia alimentar), que evidenciaram redução da toxicidade das soluções de TOH e BPA após tratamento, sendo que o mesmo não ocorreu para o resíduo de timol, provavelmente devido à sua constituição. Os resultados indicam que o tempo de tratamento seria uma variável importante a ser estudada no tratamento dos resíduos de TOH gerados nos laboratórios do CENA/USP. Os POA (O3/UV e O3/UV/H2O2) propostos no presente trabalho mostraram-se uma alternativa adequada, eficiente e com custos competitivos em relação a outras tecnologias no tratamento destes compostos fenólicos / In order to facilitate, environmentally and economically the mineralization of phenolic substances present in water and wastewater, a study was carried out with the molecules of thymol (C10H14O TOH, in a concentration of 200 mg L-1) and bisphenol-A (C15H16O2 BPA, in a concentration of 25 mg L-1) using Advanced Oxidation Processes (AOP). The AOP used were ozone (O3), ultraviolet radiation (UV) and hydrogen peroxide (H2O2) which, when combined or in specific conditions, promote a hydroxyl radicals (OH) formation, a powerful oxidant. The line system used was developed in the Residues Treatment Laboratory at CENA/USP. The TOH and BPA were submitted to the treatment at pH 3 and 11, with O3 (1.98 g h-1) and UV (0.8 and 32 watts). Additionally, the TOH was also treated with H2O2 (0, 1,176 and 2,352 mg), totalizing 18 treatments for TOH and 6 for BPA. The treatments efficiency was assessed by the percentage of total organic carbon (TOC) removed. For both molecules, the combination of 1.98 g h-1 of O3, 32 watts of UV and absence of H2O2, at pH 3, showed the best conversion of TOC to CO2, specifically 99.6 ± 0.6% of TOH in 90 minutes and 93.9 ± 0.6% of BPA in 60 minutes of treatment. This treatment was reproduced in the TOH residue generated by the CENA/USP laboratories. The TOC removal was 92.6 ± 1.2%, slightly lower than that obtained for the solution of TOH, probably due to the presence of other substances. The solutions (BPA and TOH) and the residue (TOH) treated were submitted to the total phenols analysis, resulting in values lower than demanded (0.5 mg L-1 of total phenols) by the law in force (CONAMA 357/2005 and 397/2008), which regulates the discharge of wastewater in the environment. We carried out also toxicological tests with the organisms Pseudokirchneriella subcapitata, Daphnia magna and Hydra attenuate (representing species in different levels of the food chain). The toxicity of solutions (TOH and BPA) decreased after treatment, but the same results were not observed in the TOH residue, probably due to its composition. The results show that treatment time is an important variable that should be studied for treatment of TOH residue generated in the CENA/USP laboratories. The AOP (O3/UV and O3/UV/H2O2) proposed showed up as a suitable alternative in terms of efficiency and cost-competitive when compared with other technologies to treatment of these phenolic compounds

Orientador: Giuseppina Pace Pereira Lima / Banca: João Domingos Rodrigues / Banca: Denise Laschi / Banca: Terezinha Rangel Câmara / Banca: José Luis Mosca / Abstract: Beehive ginger inflorescences have yellow bracts when they are young, which are ornamental and are specially used in gardening projects and as cut flowers. However, there are crop management and postharvest factors that affect the expansion of the species. So, the objective of this work was to study some physiological postharvest aspect of beehive ginger inflorescences grown in the lower middle São Francisco river basin. Flower stems just harvested were submitted to different treatments (distilled water; 75 mg L-1 of silver nitrate - AgNO3; 1000 mg L-1 of cobalt chloride - CoCl2; 5 mg L-1 de GA3 - Progibb® and 10 mg L-1 of 6-benzylamino purine - BAP), in an environment with controlled temperature and humidity, for 15 days. The longevity was monitored from non-destructive analysis (grading scale, fresh weight, consumption and pH of the preservative solution) as well as destructive ones (peroxidase and polyphenol oxidase activity, total soluble and reducing carbohydrates, phenols, putrescine, spermine and spermidine content). The non-destructive analysis showed that the beehive ginger stems treated with gibberelin and silver nitrate presented a better visual aspect according to the grading scale, the ones treated with AgNO3 absorbed a greater volume of the solution during the experimental period, while the ones treated with silver nitrate and BAP had a greater fresh weight. The smallest variation of the preservative solution pH took place with the treatments containing the plant regulators. The destructive analysis revealed that the inflorescences maintained in preservative solutions with gibberelin and distilled water kept their stocks of total soluble sugars for 3 days longer than the stems submitted to the other treatments. The contents of reducing sugars increased 7 considerably in inflorescences treated with cytokinin. The BAP promoted alterations in the activity of the... (Complete abstract click electronic access below) / Doutor

Permafrost soils in the Arctic store large quantities of organic matter, roughly twice the amount of carbon that was present in the atmosphere before the industrial revolution. This freeze-locked carbon pool is susceptible to thawing caused by amplified global warming at high latitudes. The remobilization of old permafrost carbon facilitates its degradation to carbon dioxide and methane, thereby providing a positive feedback to climate change. Accelerating coastal erosion in addition to projected rising river discharge with enhancing sediment loads are anticipated to transport increasing amounts of land-derived organic carbon (OC) to the Arctic Ocean. On its shallow continental shelves, this material may be remineralized in the water column or in the sediments, transported without being altered off shelf towards the deep sea of the Arctic Interior or buried in marine sediments and hence sequestered from the contemporary carbon cycle. The fate of terrigenous material in the marine environment, though offering potentially important mechanisms to either strengthen or attenuate the permafrost-carbon climate feedback, is so far insufficiently understood. In this doctoral thesis, sediments from the wide East Siberian Arctic Shelf, the world’s largest shelf-sea system, were used to investigate some of the key processes for OC cycling. A range of bulk sediment properties, carbon isotopes and molecular markers were employed to elucidate the relative importance of different organic matter sources, the role of cross-shelf transport and the relevance of degradation during transport and after burial. Overall, OC released from thawing permafrost constitutes a significant proportion of the sedimentary organic matter on the East Siberian Arctic Shelf. Two sediment cores from the inner and outer East Siberian Sea recorded no substantial changes in source material or clear trends in degradation status for the last century. With increasing distance from the coast, however, strong gradients were detected towards lower concentrations of increasingly reworked land-derived OC. The time spent during cross-shelf transport was consequently found to exert first-order control on degradation. Compound-specific radiocarbon dating on terrigenous biomarkers revealed a net transport time of ~4 000 years across the 600 km wide Laptev Sea shelf, yielding degradation rate constants for bulk terrigenous OC and specific biomarkers on the order of 2-4 kyr-1. From these results, the carbon flux released by degradation of terrigenous OC in surface sediments was estimated to be ~1.7 Gg yr-1, several orders of magnitude lower than what had been quantified earlier for dissolved and particulate OC in the water column. Lower oxygen availability and close associations with the mineral matrix may protect sedimentary OC from remineralization and thereby weaken the permafrost-carbon feedback to present climate change. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Submitted. Paper 4: Manuscript.</p>

Com o intuito de viabilizar, ambiental e economicamente, a mineralização de substâncias fenólicas presentes em águas e efluentes, foram estudadas as moléculas do timol (C10H14O TOH, a 200 mg L-1) e do bisfenol-A (C15H16O2 BPA, a 25 mg L-1) utilizando Processos Oxidativos Avançados (POA), no caso, ozônio (O3), radiação ultravioleta (UV) e peróxido de hidrogênio (H2O2) que, quando combinados, ou em condições específicas, promovem a geração de radicais hidroxila (OH), um poderoso oxidante. O sistema de tratamento utilizado foi desenvolvido no próprio Laboratório de Tratamento de Resíduos do CENA/USP. O TOH e o BPA foram submetidos a tratamento em pH 3 e 11, com O3 (1,98 g h-1) e UV (0, 8 e 32 watts). Adicionalmente, o TOH foi tratado também com H2O2 (0, 1.176 e 2.352 mg), totalizando 18 tratamentos para o TOH e 6 para o BPA. A eficiência dos tratamentos foi avaliada a partir da porcentagem de remoção do carbono orgânico total (TOC). Para ambas as moléculas, a combinação 1,98 g h-1 de O3, 32 watts UV e ausência de H2O2, em pH 3, apresentou os melhores resultados de conversão de TOC a CO2, especificamente 99,6 ± 0,6% em 90 minutos para o timol e 93,9 ± 0,6% de mineralização de bisfenol-A em 60 minutos de tratamento. Este tratamento foi então reproduzido no resíduo de timol gerado pelos laboratórios do CENA/USP e o resultado obtido foi de 92,64 ± 1,2%, um pouco inferior ao obtido para a solução de TOH, provavelmente devido à presença de outras substâncias. As soluções (TOH e BPA) e o resíduo (TOH) tratados foram submetidos à análise de fenóis totais, obtendo-se resultados inferiores ao exigido (0,5 mg L-1 de fenóis totais) pelas resoluções CONAMA 357/2005 e 397/2008 para a disposição em corpos hídricos. Realizaram-se também ensaios de toxicidade com os organismos Pseudokirchneriella subcapitata, Daphnia magna e Hydra attenuata (que representam os distintos níveis tróficos da cadeia alimentar), que evidenciaram redução da toxicidade das soluções de TOH e BPA após tratamento, sendo que o mesmo não ocorreu para o resíduo de timol, provavelmente devido à sua constituição. Os resultados indicam que o tempo de tratamento seria uma variável importante a ser estudada no tratamento dos resíduos de TOH gerados nos laboratórios do CENA/USP. Os POA (O3/UV e O3/UV/H2O2) propostos no presente trabalho mostraram-se uma alternativa adequada, eficiente e com custos competitivos em relação a outras tecnologias no tratamento destes compostos fenólicos / In order to facilitate, environmentally and economically the mineralization of phenolic substances present in water and wastewater, a study was carried out with the molecules of thymol (C10H14O TOH, in a concentration of 200 mg L-1) and bisphenol-A (C15H16O2 BPA, in a concentration of 25 mg L-1) using Advanced Oxidation Processes (AOP). The AOP used were ozone (O3), ultraviolet radiation (UV) and hydrogen peroxide (H2O2) which, when combined or in specific conditions, promote a hydroxyl radicals (OH) formation, a powerful oxidant. The line system used was developed in the Residues Treatment Laboratory at CENA/USP. The TOH and BPA were submitted to the treatment at pH 3 and 11, with O3 (1.98 g h-1) and UV (0.8 and 32 watts). Additionally, the TOH was also treated with H2O2 (0, 1,176 and 2,352 mg), totalizing 18 treatments for TOH and 6 for BPA. The treatments efficiency was assessed by the percentage of total organic carbon (TOC) removed. For both molecules, the combination of 1.98 g h-1 of O3, 32 watts of UV and absence of H2O2, at pH 3, showed the best conversion of TOC to CO2, specifically 99.6 ± 0.6% of TOH in 90 minutes and 93.9 ± 0.6% of BPA in 60 minutes of treatment. This treatment was reproduced in the TOH residue generated by the CENA/USP laboratories. The TOC removal was 92.6 ± 1.2%, slightly lower than that obtained for the solution of TOH, probably due to the presence of other substances. The solutions (BPA and TOH) and the residue (TOH) treated were submitted to the total phenols analysis, resulting in values lower than demanded (0.5 mg L-1 of total phenols) by the law in force (CONAMA 357/2005 and 397/2008), which regulates the discharge of wastewater in the environment. We carried out also toxicological tests with the organisms Pseudokirchneriella subcapitata, Daphnia magna and Hydra attenuate (representing species in different levels of the food chain). The toxicity of solutions (TOH and BPA) decreased after treatment, but the same results were not observed in the TOH residue, probably due to its composition. The results show that treatment time is an important variable that should be studied for treatment of TOH residue generated in the CENA/USP laboratories. The AOP (O3/UV and O3/UV/H2O2) proposed showed up as a suitable alternative in terms of efficiency and cost-competitive when compared with other technologies to treatment of these phenolic compounds

Óleos contendo alta proporção de ácidos graxos ômega-3 (n-3 FA) têm sido aplicados na formulação de alimentos ou comercializados como suplementos, com a finalidade de reduzir o risco cardiovascular, principalmente devido aos seus efeitos hipotriglicêmicos e anti-inflamatórios. No entanto, a susceptibilidade à oxidação dos n-3 FA é elevada, levando à formação de vários produtos secundários, incluindo alguns tóxicos e potencialmente aterogênicos. Por esta razão, compostos naturais com propriedades antioxidantes têm sido investigados com o objetivo de melhorar a estabilidade oxidativa dos óleos com alta proporção de n-3 FA. Neste estudo, avaliou-se a capacidade antioxidante de dois compostos naturais (ácido sinápico e hidrato de rutina) utilizando-se um modelo acelerado para oxidar os óleos. Foram combinados cinco indutores (Temperatura; Ferro- Fe2+; 2,2\'-Azobis dicloridrato de 2-amidinopropano - AAPH; ascorbil palmitato - AP e 2,2\'-azobis -2,4-dimetilvaleronitrilo - AMVN) em um delineamento fatorial (25-1) com ½ fração de \"resolução V\" para acelerar a oxidação de três óleos (linhaça, Echium e peixe) contendo diferentes fontes de n-3 FA: ácido &#945;-linolênico (ALA), ácido estearidônico (SDA) e ácido eicosapentaenóico (EPA) + ácido docosahexaenóico (DHA), respectivamente. Não houve diferença entre os marcadores de oxidação (LOOH e TBARS) estimados pelos modelos e os valores observados experimentalmente. Os indutores AMVN e Fe2+ foram os principais fatores responsáveis pelo aumento da concentração de TBARS. Os valores dos marcadores oxidativos obtidos 48 h após a indução foram semelhantes ou superiores àqueles observados nas amostras oxidadas a 60°C por 15 dias, sendo ambos maiores que os valores observados nas amostras de óleo frescas. Entre os compostos voláteis formados pela oxidação de diferentes fontes de n-3 FA, (E, E) 2,4 -heptadienal, (E, E) 2,4-decadienal, decanal, undecanal e (E) -2-undecenal foram identificados em todas as amostras, podendo ser utilizados como marcadores oxidativos mais específicos. Utilizando o modelo de oxidação acelerada, o hidrato de rutina melhorou a estabilidade oxidativa do óleo de peixe, provavelmente devido à presença de grupos catecol em sua estrutura química. Este estudo contribuiu para que ensaios mais rápidos fossem realizados na avaliação do efeito antioxidante de novas moléculas aplicadas em óleos funcionais comestíveis. / Oils containing a high proportion of omega-3 fatty acids (n-3 FA) have been used in the formulation of foods or sold as supplements, aiming to reduce cardio-vascular risks, mainly due to their hypotriglycemic and anti-inflammatory effects. However, n-3 FA are highey susceptible to oxidation, leading to the formation of several products, including some toxic and potentially atherogenic. For this reason, natural products with antioxidant properties have been investigated to improve the oxidative stability of oils with a high proportion of n-3 FA. This study evaluated the antioxidant capacity of two natural compounds (sinapic acid and rutin hydrate), using an accelerated model to oxidize the oils. Five inducers were combined (Temperature, Iron-Fe2+, 2,2\'-Azobis (2-amidinopropane) dihydrochloride-AAPH, Ascorbyl palmitate-AP and 2,2\'-azobis-2,4-dimethylvaleronitrile-AMVN) in a factorial design (25-1) ½ fraction of \"resolution V\" to accelerate the oxidation of three oils (flaxseed, Echium and fish) containing different sources of n-3 FA: &#945;-linolenic acid (ALA), stearidonic acid (SDA) and eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA), respectively. There was no difference between the oxidation markers (LOOH and TBARS) estimated by the regression models and the values experimentally observed. The inducers AMVN and Fe2+ were the main factors responsible for the increase of TBARS concentration. The values of the oxidation markers obtained 48h after the induction were similar to or higher than those obtained when the samples were oxidized at 60°C for 15 days, both being more elevated than the values observed in the fresh oils. Among the volatile compounds formed by the oxidation of different sources of n-3 FA, (E, E) 2,4-heptadienal, (E, E) 2,4-decadienal, decanal, undecanal and (E)-2-undecenal were identified in all samples, and could be used as more specific oxidation markers. Using the accelerated model, rutin hydrate improved the oxidative stability of fish oil, probably due to the presence of catechol groups in its chemical structure. This study showed that faster anays could be performed to evaluate the antioxidant effect of new molecules applied on edible functional oils.

<p>Болести представљају један од најважнијих фактора који утичe на успешност производње сунцокрета. Црна пегавост стабла сунцокрета, коју проузрокује патогена гљива Phoma macdonaldii, представља болест која у појединим регионима распрострањења значајно смањује принос и квалитет сунцокрета. Ранија истраживања спроведена у агроеколошким условима Србије су идентификовала ову болест као широко распрострањену. Истовремено је утврђено да знатан број генотипова сунцокрета био осетљив.<br />Циљ рада је био да се установи степен отпорности инбред линија и хибрида према овој болести као и постојање извора отпорности код популација врста из рода Helianthus. Вишегодишње праћење болести на више локалитета и на већем броју хибрида за циљ је имало одређивање утицаја генотипа и спољашње средине на појаву и развој болести. Тестирана је могућност кориштења ергостерола као индикатора постојања отпорности. Праћењем концентрације фенолних једињења тестирана је реакција домаћина на присуство патогена са акцентом на период непосредно након инокулације. Други део истраживања је био фокусиран на патогена са циљем утврђивања варијабилности популације кориштењем различитих метода. Утврђена је агресивност изолата и генетска варијабилност патогена помоћу RAPD маркера. Постојање варијабилности је проверено и помоћу одгајивачких особина.<br />Тестирањем генотипова сунцокрета, применом метода вештачке инокулације у току четворогодишњег периода, утврђено је постојање статистички значајних разлика између генотипова. Од седамдесет инбред линија отпорност на највећем броју локалитета где је вршено тестирање испољиле су линије DOP-32-08, Ph-BC1-162, Ph-BC1-53, IMI-AB-12, CMS-3-8, BT-VL-2, DOP-33-08, BT-VL-24 и OD-DI-98. Реакција хибрида је варирала у односу на годину тестирања. Највећи степен отпорности су испољили хибриди Баћа и НС Брилијант. Смањење у напредовању болести у години са израженим дефицитом у падавинама је забележено код линија гајених у условима сувог ратарења. Тестирањем 181 популације 9 једногодишњих врста рода Helianthus у условима природне инфекције, утврђене су популације код свих одабраних врста код којих болест није била присутна. Утврђивањем количине ергостерола у оквиру симптома болести код биљака инбред линија у контролисаним условима утврђено је повећање количина овог једињења код осетљиве инбред линије у поређењу са линијама са одређеним степеном отпорности. Резултатима гајења 16 хибрида на већем броју локалитета у току четири године, установљено је постојање значајног утицаја средине и генотипа на појаву и интензитет<br />болести. Највећу отпорност у природним условима су испољили хибриди Баћа, НС-Х-111 и НС Фантазија. Гајењем биљака у условима различитог времена сетве, појава и интензитет црне пегавости стабла су имали тенденцију опадања са каснијом сетвом. Примена само азотних ђубрива највише је допринела порасту појаве и интензитета болести, а исти параметри су били највећи код најмање густине гајења биљака. Анализом фенолних једињења у ткиву листа, у одређеним временским интервалима, након вештачке инокулације биљака инбред линија са различитим степеном осетљивости, утврђен је повећан садржај хлорогене киселине у току прва 24 сата код узорака листа отпорне линије. Четири линије различитог степена осетљивости су инокулисане у контролисаним условима са различитим изолатима патогена. Установљено је постојање варијабилности у погледу агресивности, док су 4 четири од укупно 56 тестираних изолата испољили висок степен агресивности. Груписање изолата сличног степена агресивности према географском пореклу није установљено. Анализом генетске варијабилности помоћу RAPD маркера утврђен је висок степен полиморфизма код употребљених изолата. Од употребљених прајмера највећу информативност је имао прајмер OPE-04. Висок степен варијабилности је потврђен кластер анализом. AMOVA анализом утврђено је да се генетска варијабилност изолата гљиве односи највећим делом на варијабилност између изолата. Генетска варијабилност између изолата груписаних по регионима и агресивности је била занемарљива.<br />Слични обрасци су запажени и након утвђивања одгајивачких особина &bdquo;in vitro&ldquo;. Најбољи раст и продукција пикнида је забележена на подлози од овса. Између изолата су забележене значајне разлике у пречнику колонија и продукцији пикнида, али те разлике нису биле повезане са географским пореклом изолата.<br />Црна пегавост сунцокрета у Србији je сталан пратилац сунцокрета. Интензит болести је значајно условљен степеном отпорности генотипова сунцокрета, агресивношћу изолата и временским условима. Патоген испољава велик степен генетске варијабилности што га чини прилагодљивим за измене у генетској структури домаћина и средини. Однос сунцокрет-Phoma macdonaldii представља динамичан систем који захтева наставак истраживања уз мониториг.</p> / <p>Bolesti predstavljaju jedan od najvažnijih faktora koji utiče na uspešnost proizvodnje suncokreta. Crna pegavost stabla suncokreta, koju prouzrokuje patogena gljiva Phoma macdonaldii, predstavlja bolest koja u pojedinim regionima rasprostranjenja značajno smanjuje prinos i kvalitet suncokreta. Ranija istraživanja sprovedena u agroekološkim uslovima Srbije su identifikovala ovu bolest kao široko rasprostranjenu. Istovremeno je utvrđeno da znatan broj genotipova suncokreta bio osetljiv.<br />Cilj rada je bio da se ustanovi stepen otpornosti inbred linija i hibrida prema ovoj bolesti kao i postojanje izvora otpornosti kod populacija vrsta iz roda Helianthus. Višegodišnje praćenje bolesti na više lokaliteta i na većem broju hibrida za cilj je imalo određivanje uticaja genotipa i spoljašnje sredine na pojavu i razvoj bolesti. Testirana je mogućnost korištenja ergosterola kao indikatora postojanja otpornosti. Praćenjem koncentracije fenolnih jedinjenja testirana je reakcija domaćina na prisustvo patogena sa akcentom na period neposredno nakon inokulacije. Drugi deo istraživanja je bio fokusiran na patogena sa ciljem utvrđivanja varijabilnosti populacije korištenjem različitih metoda. Utvrđena je agresivnost izolata i genetska varijabilnost patogena pomoću RAPD markera. Postojanje varijabilnosti je provereno i pomoću odgajivačkih osobina.<br />Testiranjem genotipova suncokreta, primenom metoda veštačke inokulacije u toku četvorogodišnjeg perioda, utvrđeno je postojanje statistički značajnih razlika između genotipova. Od sedamdeset inbred linija otpornost na najvećem broju lokaliteta gde je vršeno testiranje ispoljile su linije DOP-32-08, Ph-BC1-162, Ph-BC1-53, IMI-AB-12, CMS-3-8, BT-VL-2, DOP-33-08, BT-VL-24 i OD-DI-98. Reakcija hibrida je varirala u odnosu na godinu testiranja. Najveći stepen otpornosti su ispoljili hibridi Baća i NS Brilijant. Smanjenje u napredovanju bolesti u godini sa izraženim deficitom u padavinama je zabeleženo kod linija gajenih u uslovima suvog ratarenja. Testiranjem 181 populacije 9 jednogodišnjih vrsta roda Helianthus u uslovima prirodne infekcije, utvrđene su populacije kod svih odabranih vrsta kod kojih bolest nije bila prisutna. Utvrđivanjem količine ergosterola u okviru simptoma bolesti kod biljaka inbred linija u kontrolisanim uslovima utvrđeno je povećanje količina ovog jedinjenja kod osetljive inbred linije u poređenju sa linijama sa određenim stepenom otpornosti. Rezultatima gajenja 16 hibrida na većem broju lokaliteta u toku četiri godine, ustanovljeno je postojanje značajnog uticaja sredine i genotipa na pojavu i intenzitet<br />bolesti. Najveću otpornost u prirodnim uslovima su ispoljili hibridi Baća, NS-H-111 i NS Fantazija. Gajenjem biljaka u uslovima različitog vremena setve, pojava i intenzitet crne pegavosti stabla su imali tendenciju opadanja sa kasnijom setvom. Primena samo azotnih đubriva najviše je doprinela porastu pojave i intenziteta bolesti, a isti parametri su bili najveći kod najmanje gustine gajenja biljaka. Analizom fenolnih jedinjenja u tkivu lista, u određenim vremenskim intervalima, nakon veštačke inokulacije biljaka inbred linija sa različitim stepenom osetljivosti, utvrđen je povećan sadržaj hlorogene kiseline u toku prva 24 sata kod uzoraka lista otporne linije. Četiri linije različitog stepena osetljivosti su inokulisane u kontrolisanim uslovima sa različitim izolatima patogena. Ustanovljeno je postojanje varijabilnosti u pogledu agresivnosti, dok su 4 četiri od ukupno 56 testiranih izolata ispoljili visok stepen agresivnosti. Grupisanje izolata sličnog stepena agresivnosti prema geografskom poreklu nije ustanovljeno. Analizom genetske varijabilnosti pomoću RAPD markera utvrđen je visok stepen polimorfizma kod upotrebljenih izolata. Od upotrebljenih prajmera najveću informativnost je imao prajmer OPE-04. Visok stepen varijabilnosti je potvrđen klaster analizom. AMOVA analizom utvrđeno je da se genetska varijabilnost izolata gljive odnosi najvećim delom na varijabilnost između izolata. Genetska varijabilnost između izolata grupisanih po regionima i agresivnosti je bila zanemarljiva.<br />Slični obrasci su zapaženi i nakon utvđivanja odgajivačkih osobina &bdquo;in vitro&ldquo;. Najbolji rast i produkcija piknida je zabeležena na podlozi od ovsa. Između izolata su zabeležene značajne razlike u prečniku kolonija i produkciji piknida, ali te razlike nisu bile povezane sa geografskim poreklom izolata.<br />Crna pegavost suncokreta u Srbiji je stalan pratilac suncokreta. Intenzit bolesti je značajno uslovljen stepenom otpornosti genotipova suncokreta, agresivnošću izolata i vremenskim uslovima. Patogen ispoljava velik stepen genetske varijabilnosti što ga čini prilagodljivim za izmene u genetskoj strukturi domaćina i sredini. Odnos suncokret-Phoma macdonaldii predstavlja dinamičan sistem koji zahteva nastavak istraživanja uz monitorig.</p> / <p>Diseases are a major constraint in sunflower production. Phoma black stem, caused by pathogenic fungus Phoma macdonaldii, is widely distributed disease known in some regions for extensive damage to sunflower yield and quality. Previous research conducted in Serbia and testing for resistance with isolates of pathogen coming from this region identitified disease as common. At the same time, reports on resistance to this disease suggested majority of tested genotipes to be susceptible.<br />Aim of this research was to quantify level of resitance in selected inbreed lines and hybrids as well as to find sources of resistance in species from genus Helianthus. Multiyear monitoring of disease on different hybrids and locations was conducted to determine significance of genotype and environment influence on incidence and severity od disease. Usage of ergosterol as indicator of resistance was also tested. Host reaction to disease was monitored by quantification of total soluble phenols and different phenolic compounds known to be impacted during disease progress in other host-pathogen systems. In this part of research emphasis was particularly on period of time following the inoculation. Secound part of research was focused on pathogen. Aim was to determine variability of pathogen population using several methods. Four inbreed lines varyng in level of susceptibility were used for determination of isolate aggressiveness and RAPD markers were used for genetic variability determination. Distinctions among isolates were define using cultural characteristics.<br />Results of sunflower genotypes survey after application of artificial inoculation methods during 4-year period confirmed presence of significant differences in resistance to phoma black stem among tested genotypes. Out of 70 tested sunflower, inbred lines with the highest level of resistance on majority of localities were DOP-32-08, Ph-BC1-162, Ph-BC1-53, IMI-AB-12, CMS-3-8, BT-VL-2, DOP-33-08, BT-VL-24 и OD-DI-98. Resistance of hybrids varied in each year with the highest level of resistance manifested by hybrids Baća and NS Brilijant. Differences between irrigated and rainfed trials were found in year characterized with drought periods after inoculation with decreased disease severity in rain fed experiment. Total of 181 populations of 9 Helianthus species was surveyed for disease incidence. Among the populations of each species ones without symptoms of disease were found. Positive correlation was found between quantity of ergosterol and level of resistance in selected sunflower inbred lines. Disease incidence and severity data of 16 hybrids from number of locations during 4-year period showed significant influence of both genotype and environment. Highest level of resistance had hybrids Baća, HS-H-111 and NS Fantazija.<br />Crop production practices influenced disease incidence and progress. Plants in experiment with different sowing times expressed different disease intensity. Both disease incidence and severity decreased along sowing timeline. Solely application of nitrogen fertilizers most enhanced disease intensity, and the same parameters were highest in plots with lowest plant density.<br />Analysis of phenolic compounds in leaf tissue revealed a significant increase in concentration of chlorogenic acid in first 24 h after inoculation of resistant genotype. In susceptible genotypes that parameter remained unchanged. Patterns of reaction of four inbred lines to disease revealed differences in aggresiveness, with 4 out of 56 tested isolates regarded as highly aggressive. Isolates with similar aggressiveness did not cluster according to geographic origin. Genetic variability analysis using RAPD markers on colected isolates revealed significant level of polymorphism, with primer OPE-04 as the most informative one. High level of genetic variability was confirmed after cluster analysis. Genetic variability among groups with similar agressiveness and geografic origin was insignificant while variability among isolates inside each group was significant according to AMOVA analysis. Similar patern was observed after determination of culturing characteristics &bdquo;in vitro&ldquo;. Pathogen growth and development was best on oat agar medium. Testing warious isolated for growth and production of picnidia revealed significant difference without corelation to geographic origin.<br />In conclusion, phoma black stem occurence on sunflower in Serbia is common. Disease incidence and severity depend on host resistance, isolate aggressiveness and environmental conditions. As a result of high level of genetic variability pathogen is addaptable to changes in host susceptibility and environment. Pathosystem sunflower-Phoma macdonaldii is highly dinamic and requires continuation of research and monitoring.</p>