Les plantes envahissantes menacent la biodiversité ainsi que les activités humaines. Afin de les maîtriser, la pulvérisation d'herbicides est une méthode fréquemment employée en Amérique du Nord. Cette approche ne fait pas toujours consensus et est même parfois interdite ou restreinte, ce qui justifie le recours à d'autres options. Les alternatives peuvent toutefois s'avérer rares, comporter d'importantes limitations ou sont peu documentées. Cette étude vise à tester l’efficacité de méthodes permettant de maîtriser et de prévenir les invasions de roseau commun (Phragmites australis), l'une des plantes envahissantes les plus problématiques sur le continent nord-américain, tout en limitant au minimum l'utilisation d'herbicides. Le potentiel de quatre méthodes de lutte aux petites populations de roseau bien établies a d'abord été évalué : l’excavation avec enfouissement sur place, le bâchage, l’excavation avec enfouissement sur place combinée au bâchage, et la fauche répétée. Les résultats ont montré que l'excavation avec enfouissement sur place, avec ou sans bâchage, a entraîné une élimination presque totale des populations visées, ce qui est comparable ou supérieur à l'effet généralement obtenu avec la pulvérisation d'herbicide. Le bâchage avec des toiles opaques, maintenues pendant un an, a pour sa part entraîné une maîtrise partielle du roseau, suggérant qu'une application prolongée serait nécessaire pour l'éradication de la plante. La fauche répétée, exécutée à raison de cinq fauches par été pendant deux ans, a fourni une efficacité mitigée. Les résultats suggèrent néanmoins que la fauche pendant plusieurs années contribue à affaiblir la plante, ce qui pourrait aider à son confinement. Une méthode additionnelle a été expérimentée afin de traiter les tiges éparses de roseau tout en limitant les risques d'effets hors cibles, soit le badigeonnage manuel d’herbicide. Suite à ces tests, les résultats ont montré une diminution importante de la densité des tiges, ce qui suggère que la méthode est efficace afin d'éliminer les repousses après un traitement initial, et pourrait également être employée sur de jeunes populations clairsemées. L'effet d'un ensemencement préventif de plantes herbacées sur l'établissement de semis de roseau a également été étudié, suite à des traitements sur de vastes parcelles de sol nu. Les résultats suggèrent que la méthode est efficace afin de limiter la propagation du roseau par semences et qu'un suivi périodique suite à l'intervention serait suffisant afin de maintenir l'effet préventif. / Invasive plants threaten biodiversity as well as human activities and in North America, attempts to control them often rely on the spraying of herbicides. This approach does not always achieve consensus and is sometimes forbidden or restricted, encouraging managers to consider other options. Alternatives can however prove to be rare, involve serious limitations or be poorly documented. This study aims at testing the efficiency of methods for the control and prevention of invasions by common reed (Phragmites australis), one of the most problematic invasive plants on the North American continent, all while reducing herbicide use to a minimum. The potential of four methods for the control of small, well established stands of common reed has first been evaluated: excavation and on-site burial, covering with heavy-duty tarps, excavation and on-site burial combined with covering with heavy-duty tarps, and repeated cutting. Results have shown that excavation with on-site burial, followed or not by covering with heavy-duty tarps, led to an almost complete elimination of the populations targeted, which is comparable or superior to the effect typically obtained with herbicide spraying. Covering common reed with opaque heavy-duty tarps, held in place for one year, lead to partial control of the plant, suggesting that holding the tarps in place for a longer period would be necessary to foresee the eradication. Cutting, repeated five times per summer for two years, gave mixed results. The results suggest, however, that cutting during multiple years contributes to weakening the plant, which could help to its confinement. An additional method, the hand-wicking of common reed stems with herbicide, has been tested to treat scattered stems while limiting off-target effects. Following these tests, the results have shown an important reduction of the density of reed stems, which suggests that the method is effective to control common reed regrowth after an initial treatment, and could be used on young stands. The effect of seeding herbaceous plants on the establishment of common reed seedlings has also been evaluated, following the sowing of vast plots of bare soil. The results suggest that the method is an efficient way of limiting the propagation of common reed through seed dispersal. Results also suggest that periodic follow-ups to eliminate seedlings would be sufficient to maintain the preventive effect.

O presente trabalho descreve o uso de eletrodos de carbono vítreo como eletrodo-base para modificação com hexacianoferratos de Fe (FeHCF), Co (CoHCF) e Ni (NiHCF) por meio da adsorção destes complexos utilizando a técnica de voltametria cíclica. Os eletrodos quimicamente modificados (EQMs) preparados por este método apresentam características diferenciadas tais como robustez e estabilidade ao longo das medidas eletroquímicas, além de boas propriedades redox. Os EQMs com FeHCF e NiHCF foram avaliados em função da sua atividade eletrocatalítica para a reação de oxidação do Ácido Ascórbico em solução de KCl 0,1 mol L-1 (pH 6,6). O EQM com CoHCF foi avaliado nas mesmas condições dos outros filmes, porém não apresentou uma diminuição no sobrepotencial de oxidação do Ácido Ascórbico. Embora não tenha apresentado uma resposta semelhante aos EQMs com FeHCF e NiHCF, o EQM com CoHCF apresentou a melhor sensibilidade da resposta de corrente de pico quando comparado aos demais. A fim de proporcionar a renovação da superfície do eletrodo foi avaliada a utilização de eletrodos de pasta de carbono modificada com FeHCF, CoHCF ou NiHCF. Os parâmetros de composição da pasta, o eletrólito suporte e o pH foram avaliados utilizando o EPCM com FeHCF por meio da técnica de voltametria cíclica e os parâmetros otimizados foram aplicados no uso dos EPCM com CoHCF e NiHCF. Para avaliar a sensibilidade na deteccção da L-Cisteína, do Glifosato e da N-Acetilcisteína, os EPCMs modificados foram submetidos a estudos com a técnica de voltametria de onda quadrada. Todos os analitos apresentaram resposta eletroquímica em todos os EPCMs, porém as melhores respostas foram para o FeHCF na presença de L-Cisteína, CoHCF na presença de Glifosato e NiHCF na presença de N-Acetilcisteína. Na presença de 1 x 10-8 mol L-1 dos analitos os EPCMs com hexacianoferratos apresentaram um boa resposta eletroquímica, embora não tenha sido possível observar uma linearidade entre o aumento da concentração dos analitos propostos e a corrente de pico. Essa sensibilidade demonstrada pelos EPCMs na presença de moléculas com grupos funcionais, como ácidos carboxílicos, aminoácidos e tióis abre novas perspectivas para a análise direta de compostos de interesse clínico, farmacêutico e amostras de interesse ambiental. / This work describes the use of glassy carbon electrodes as the base for electrode modification with hexacyanoferrates of Fe (FeHCF), Co (CoHCF) and Ni (NiHCF) through adsorption of these complexes using cyclic voltammetry. The chemically modified electrodes (CMEs) prepared by this method have specific features such as robustness and stability along the electrochemical measurements, and good redox properties. The CME with FeHCF and NiHCF were evaluated take into accounting their electrocatalytic activity for oxidation reaction of Ascorbic Acid in KCl 0.1 mol L-1 (pH 6.6). The CME with CoHCF was evaluated under the same conditions, but did not show a decrease on the overpotential for Ascorbic Acid oxidation. Although the CME with CoHCF has not showed the same behavior presented by CMEs with FeHCF and NiHCF, the CME with CoHCF presented the best sensitivity in order to relationship between peak current and concentration of Ascorbic Acid when compared to CMEs with either. In order to provide the renewal of the electrode surface was evaluated the use of carbon paste electrodes (CPEs) modified with FeHCF, CoHCF and NiHCF. The CPE parameters such as composition, supporting electrolyte and pH were evaluated using the CPE with FeHCF by cyclic voltammetry and these optimized parameters were applied for CPEs with CoHCF and NiHCF. The studies about the sensitivity of the detection for L-Cysteine, glyphosate and N-acetylcysteine using the modified CPEs with hexacyanoferrates were performed by square wave voltammetry. All analytes showed electrochemical response at all CPEs, but the best electrochemical responses were obtained for FeHCF in the presence of L-Cysteine, CoHCF in the presence of glyphosate and NiHCF in the presence of N-acetylcysteine. The CPEs with hexacyanoferrates in the presence of 1 x 10-8 mol L-1 of L-Cysteine, glyphosate or N-acetylcysteine had showed a good electrochemical response, although were not possible to observe a linear correlation between the concentration of analytes and peak current. This sensitivity demonstrated by CPEs with hexacyanoferrates electrodes in presence of specific functional groups such as carboxylic acids, amino acids and thiols opens new perspectives for the direct analysis of compounds with clinical, pharmaceutical and environmental samples.

Surfactants are one of the more ubiquitous contaminants in aquatic systems. Their importance as toxic components of pesticide formulations has, however, been largely overlooked. Amphibians particularly, as inhabitants of shallow, temporary and often lentic aquatic environments may be at risk from exposure to these chemicals when they enter aquatic systems. This thesis presents data on the toxicity of surfactants to amphibians. Several experimental exposures were conducted with embryo-larval, tadpole and adult developmental stages of the Australian species- Crinia insignifera, Helcioporus eyrei, Limnodynastes dorsalis and Litoria moorei and the exotic species- Bufo marinus and Xenopus laevis. Animals were variously exposed to glyphosate formulations that contain a high proportion of nonionic surfactants, or commercial pesticide wetting agents (alcohol alkoxylate and nonylphenol ethoxylate (NPE) surfactants). Feeding stage tadpoles of C.insignifera, H. eyrei, L. dorsalis and L. moorei were exposed to three commercial glyphosate formulations, glyphosate isopropylamine and glyphosate acid in static-renewal acute toxicity tests. The 48-h LC50 values for Roundup Herbicide (MON 2139) tested against tadpoles of C. insignifera, H. eyrei, L. dorsalis and L. moorei ranged between 8.1 and 32.2 mg/L (2.9 and 11.6 mg/L glyphosate acid equivalent (ae)), while the 48-h LC50 values for Roundup Herbicide tested against adult and newly metamorphosed C. insignifera ranged from 137-144 mg/L (49.4-51.8 mg/L ae). / Touchdown Herbicide (4 LC-E) tested against tadpoles of C. insignifera, H. eyrei, L. dorsalis and L. moorei was slightly less toxic than Roundup with 48-h LC50 values ranging between 27.3 and 48.7 mg/L (9.0 and 16.1 mg/L ae). Roundup Biactive (MON 77920) was practically non-toxic to tadpoles of the same four species producing 48-h LC50 values of 911 mg/L (328 mg/L ae) for L. moorei and >1000 mg/L (>360 mg/L ae) for C. insignifera, H. eyrei and L. dorsalis. Glyphosate isopropylamine was practically non-toxic producing no mortality amongst tadpoles of any of the four species over 48 h, at concentrations between 503 and 684 mg/L (343 and 466 mg/L ae). The toxicity of technical grade glyphosate acid (48-h LC50, 81.2-121 mg/L) is likely to be due to acid intolerance. Feeding stage tadpoles of B. marinus, X laevis, C. insignifera, H.eyrei, L. dorsalis and L. moorei were exposed to NPE and alcohol alkoxylate in static renewal acute toxicity tests. All species exhibited non-specific narcosis following exposure to both these surfactants. The 48-h EC50 values for NPE ranged between 1.1 mg/L (mild narcosis) and 12.1 mg/L (full narcosis). The 48-h EC50 values for alcohol alkoxylate ranged between 5.3 mg/L (mild narcosis) and 25.4 mg/L (full narcosis). Xenopus laevis was the most sensitive species tested. The sensitivity of the other five species was size dependent with larger species displaying greater tolerance. Replicate acute toxicity tests with B. marinus exposed to NPE at 30 degrees celsius over 96 hours indicated that the narcotic effects were not particularly time dependant. / The mean 24, 48, 72 and 96-h EC50 (mild narcosis) were 3.6, 3.7, 3.5 and 3.5 mg/L respectively. The mean 24, 48, 72, and 96-h EC50 (full narcosis) values were 4.0, 4.1, 4.2 and 4.0 respectively. Acute toxicity tests with B. marinus exposed to NPE at 30 degrees celsius under conditions of low dissolved oxygen (0.8-2.3 mg/L) produced a two to threefold increase in toxicity. The 12-h EC50 values ranged from 1.4 to 2.2 mg/L. The embryotoxicity of NPE was determined in X. laevis, L. adelaidensis and C. insignifera using a Frog Embryo Teratogenesis assay-Xenopus (FETAX). The 96-h LC50, EC50 and MCIG (LOEC) values for X. laevis were 3.9 to 5.4 mg/L, 2.8 to 4.6 mg/L and 1.0 to 3.0 mg/L respectively. The 140-h LC50, ECSO and MCIG values for L. adelaidensis were 9.2 mg/L, 8.8 mg/L and 5.1 to 6.0 ing/L respectively. The 134-h LC50, EC50 and MCIG values for C. insignifera were 6.4 mg/L, 4.5 mg/L and 4.0 mg/L respectively. Teratogenicity indices for the three species ranged between 1.0 and 1.6 indicating either no or low teratogenicity. Xenopus laevis was the more sensitive of the three species and the only species that displayed indisputable terata. The acute toxicity data indicated that the amphibian species tested were of similar sensitivity to fish and some invertebrates. Developmental retardation and oestrogenic effects following exposure to nonylphenol ethoxylate were indicated by sublethal toxicity tests. Crinia insignifera embryos were exposed during early embryogenesis to sublethal concentrations of NPE. / Exposure to NPE did not affect either weight nor size (snout-vent length) at metamorphosis. Exposure to 5.0mg/L NPE resulted in a significant delay in the time required to reach metamorphosis. Also, exposure to 3.0 mg/L NPE for the first 6 days of embryonic development or exposure to 5.0 mg/L NPE from day 2 to day 6 resulted in a statistically significant predominance in the female phenotype amongst metamorphosing froglets. Exposure for the first five days to 1.5 ing/L or 3.0 mg/L NPE had no effect on sex ratio. The results indicated that exposure to NPEs has endocrine disruptive effects in this species and that a narrow window of susceptibility exists for the induction of predominantly female phenotype. This study has also followed the degradation of a mixture of NPE oligomers and the concomitant formation of individual oligomers in static die-away tests with and without illumination in freshwater. Over 33 days in darkness there was a progressive and complete loss of long chain oligomers (NPEO(subscript)8-17), transient increases and subsequent loss of short to medium chain oligomers (NPE0(subscript)4-7), and large persistent increases (approximately 1000%) in short chain oligomers (NPE0(subscript)1-3). In the presence of illumination, biodegradation was retarded and heterotrophic bacterial proliferation was inhibited. After 33 days there was complete loss of long chain oligomers (NPE0(subscript)9-17), incomplete loss of medium chain oligomers (NPE0(subscript)6.8) and increases in short chain oligomers (NPE0(subscript)1-5). / This thesis discusses the importance of persistent metabolites of NPE degradation as it pertains to the habitat, developmental time frame and ecology of amphibians. Degradation of NPE is likely to occur over a time frame that is longer than that required for complete embryogenesis and metamorphosis of many species of amphibians, and may easily encompass those critical stages of development during which oestrogenic metabolites can affect development.

A series of experiments was carried out on cultivated oat (Avena sativa L. cv Amuri) to examine the efficacies of fluazifop-butyl and glyphosate against water stressed plants, plants grown in low and high nitrogen and plants treated with gibberellic acid (GA₃). Avena sativa L. was used as a test plant and on completion of the experiments, further studies were carried out on the weed species wild oat (Avena fatua L.). In the laboratory, plants maintained at wilting point for five days before and nine days after spraying and treated with fluazifop-butyl (0.5 kg a.i./ha) appeared healthy 32 days after herbicide application, while plants supplied with water throughout the experiment were completely chlorotic/necrotic and had main stem detachment from within the leaf sheaths. In the field, plants maintained unirrigated until 14 days after spraying with fluazifop-butyl (0.25 kg a.i./ha) or glyphosate (0.18 kg a.i./ha) showed greater tolerance to the herbicides than plants irrigated regularly. Values for seed head yield for water stressed and irrigated plants, 45 days after applying fluazifop-butyl, were 66 g and zero g dryweight/m² respectively. Comparable values for glyphosate treated plants were 65 g and 25 g dryweight/m². Radiolabel studies indicated that in comparision with well watered plants, water stressed plants absorbed 20% less applied ¹⁴C-glyphosate. In addition, the proportion of absorbed ¹⁴C-glyphosate translocated from the treated leaf was 15% less under water stress conditions. Uptake of ¹⁴C-fluazifop-butyl was similar under well watered and water stress conditions and was 30-40% of that applied. The proportion of absorbed ¹⁴C-activity which was transported was very low, but was greater under well watered conditions (7.6%) than under water stress conditions (4.4%). Under well watered conditions in the laboratory and field, fluazifop-butyl (0.25 kg a.i./ha) and glyphosate (0.18 kg a.i./ha) were less toxic at low nitrogen than high nitrogen. For example, 34 days after spraying with fluazifop-butyl under laboratory conditions total plant dry weight was 1.51 g and 0.56 g at 1.0 mol/m³ and 10 mol/m³ applied nitrate respectively. As with soil water content, soil nitrogen content had no effect on uptake of fluazifop-butyl. However, the proportion of absorbed fluazifop-butyl which was translocated out of the treated lamina was greater under high nitrogen conditions (26.1 %) than under low nitrogen conditions (9.3%). Under laboratory conditions, addition of 200 µg GA₃to the leaf sheaths two days prior to spraying with fluazifop-butyl or glyphosate increased the efficacy of both herbicides at low nitrogen. Similarly, under field conditions application of GA₃ (0.21 kg/ha) two days prior to spraying with glyphosate increased the performance of the herbicide against Avena sativa L. growing in a nitrogen depleted soil. At harvest, seed head yield for GA₃ treated and non-treated plants was zero and 7.4 g dry weight/m² respectively. Experiments with Avena latua L. showed that this species was tolerant of fluazifop-butyl and glyphosate when grown in low water or low nitrogen conditions. Under water stress conditions, pre-treatment with GA₃ increased the phytotoxicity of fluazifop-butyl to Avena latua L. Similarily, GA₃ enhanced the phytotoxicity of glyphosate to Avena latua L. grown under low nitrogen conditions. Reduced performance of fluazifop-butyl under stress conditions involves a reduction in translocation of herbicide to meristems, but other factors are likely to be involved. It was concluded that for glyphosate, reductions in uptake and translocation of the herbicide are important factors causing reduced performance of this herbicide under stress conditions. Possible reasons for GA₃ enhancement of fluazifop-butyl and glyphosate activity under stress conditions are discussed and the potential of growth regulators as adjuvants is considered.

The chemistry of mineral surfaces is of great importance in many different areas including natural processes occurring in oceans, rivers, lakes and soils. Manganese (hydr)oxides are one important group to these natural processes, and the thermodynamically most stable trivalent manganese (hydr)oxide, manganit (γ-MnOOH), is studied in this thesis. This thesis summarises six papers in which the surface chemistry of synthetic manganite has been investigated with respect to surface acid-base properties, dissolution, and adsorption of Cd(II) and the herbicide N-(phosphonomethyl)glycine (glyphosate, PMG). In these papers, a wide range of analysis techniques were used, including X-ray photoelectron spectroscopy (XPS), extended X-ray absorption fine structure (EXAFS) spectroscopy, Fourier transform infra-red (FTIR) spectroscopy, atomic force microscopy (AFM), scanning electron microscopy (SEM), X-ray diffraction (XRD), potentiometry, electrophoretic mobility measurements and wet chemical techniques, in order to obtain a more complete understanding of the different processes occurring at the manganite-water interface. From the combined use of these techniques, a 1-pKa acid-base model was established that is valid at pH>6. The model includes a Na+ interaction with the surface: =MnOH2+½ --> =MnOH-½ + H+ log β0 (intr.) = -8.20 = -pHiep =MnOH2+½ + Na+ --> =MnOHNa+½ + H+ log β0 (intr.) = -9.64 At pH<6 the manganite crystals dissolve and disproportionate into pyrolusite (β-MnO2) and Mn(II)-ions in solution according to: 2 γ-MnOOH + 2H+ --> β-MnO2 + Mn2+ + 2H2O log K0 = 7.61 ± 0.10 The adsorption and co-adsorption of Cd(II) and glyphosate at the manganite surface was studied at pH>6. Cd(II) adsorption displays an adsorption edge at pH~8.5. Glyphosate adsorbs over the entire pH range, but the adsorption decreases with increasing pH. When the two substances are co-adsorbed, the adsorption of Cd(II) is increased at low pH but decreased at high pH. The adsorption of glyphosate is increased in the entire pH range in the presence of Cd(II). From XPS, FTIR and EXAFS it was found that glyphosate and Cd(II) form inner sphere complexes. The binary Cd(II)-surface complex is bonded by edge sharing of Mn and Cd octahedra on the (010) plane of manganite. Glyphosate forms inner-sphere complexes through an interaction between the phosphonate group and the manganite surface. The largest fraction of this binary glyphosate complex is protonated throughout the pH range. A ternary surface complex is also present, and its structure is explained as type B ternary surface complex (surface-glyphosate-Cd(II)). The chelating rings between the Cd(II) and glyphosate, found in aqueous complexes, are maintained at the surface, and the ternary complex is bound to the surface through the phosphonate group of the ligand.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Aiming to characterize the main transgenic soybeans cultivars used in Rio Grande do Sul, experiments were conducted during the 2007/2008 season, in Santa Maria/RS. In the field, the cultivars Relmo Anta RR 82, A 6001 RG, A 8000 RG, FUNDACEP 53 RR, FUNDACEP 54 RR, FUNDACEP 56 RR, BRS 244 RR, and CD 214 RR CD 219 RR, drilled at seeds rates of 250, 400 and 550 thousand seed ha-1 and at row spacing of 0.30, 0.45 and 0.60m. The variables were: grain yield, yield components, distribution of income in different strata of the plant, morphological characteristics, rate of soil covering and amount of light incident on the canopy. The experimental design was a randomized blocks, with 4 repetitions in trifatorial outline (9 cultivars x 3 seed rates x 3 row spacing). In laboratory, tests of peroxidase reaction had been carried through only using the tegument of the seeds or the entire seeds and also seedling color for 50 soybean cultivars. For the grain yield cultivars CD 214 RR and FUNDACEP 53 RR had the best results (3589 and 3530 kg ha-1, respectively). For the distribution of income in the plant strata, there is interaction between density of planting and row spacing, which indicate the trends for each cultivar. For the amount of light incident on the canopy, the interactions between the factors of planting density, row spacing, timing of the cycle of development and level of the canopy, combined with the architecture of each cultivar define the amount of light intercepted, after flowering. The morphological response of each cultivar is different for every situation tested, indicating the need for understanding the behavior of each material, to select the practices to maximize the yield. The use of the test of reaction of peroxidase used the entire seed can be used of so efficient form how much the traditionally used method (tegument). The identification of the seedling color may be an early way to identify varietal blend. / Com o objetivo de caracterizar as principais cultivares transgênicas de soja utilizadas no Rio Grande do Sul, foram realizados experimentos durante a safra agrícola 2007/2008, em Santa Maria/RS. Em campo, as cultivares Relmo Anta 82 RR, A 6001 RG, A 8000 RG, FUNDACEP 53 RR, FUNDACEP 54 RR, FUNDACEP 56 RR, BRS 244 RR, CD 214 RR e CD 219 RR foram submetidas às densidade de semeadura de 250, 400 e 550 mil semente ha-1 e aos espaçamentos entre linhas de 0,30, 0,45 e 0,60m. As variáveis avaliadas foram: rendimento de grãos, componentes do rendimento, distribuição do rendimento em diferentes estratos da planta, características morfológicas, taxa de cobertura do solo e quantidade de luz incidente no dossel. O delineamento experimental utilizado foi o de blocos ao acaso, com 4 repetições, no esquema trifatorial (9 cultivares x 3 densidades de semeadura x 3 espaçamentos entre linhas). Em laboratório, foram realizados testes de reação da peroxidase utilizando somente o tegumento das sementes ou as sementes inteiras e também coloração do hipocótilo para 50 cultivares de soja. Para o rendimento de grãos as cultivares CD 214 RR e FUNDACEP 53 RR apresentaram os melhores resultados (3589 e 3530 kg ha-1, respectivamente). Para a distribuição do rendimento nos estratos da planta, existe interação entre densidade de semeadura e espaçamento entre linhas, os quais indicam as tendências para cada cultivar. Para a quantidade de luz incidente no dossel, as interações entre os fatores densidade de semeadura, espaçamento entre linhas, época do ciclo de desenvolvimento e nível do dossel, aliados à arquitetura de cada cultivar definem a quantidade de luz interceptada, após o florescimento. A resposta morfológica de cada cultivar é diferenciada para todas as situações testadas, indicando a necessidade do conhecimento do comportamento de cada material, para a escolha das práticas que visem maximizar o rendimento de grãos. O uso do teste de reação da peroxidase utilizando a semente inteira pode ser utilizado de forma tão eficiente quanto o método tradicionalmente utilizado (tegumento). A identificação da coloração do hipocótilo pode ser uma forma precoce de identificar mistura varietal.

Este trabalho teve como objetivo, desenvolver um modelo molecular que possa descrever propriedades de equilíbrio, bem como as forças de interação envolvidas em um sistema enzima-inibidor, onde a enzima é a 5-Enolpiruvil-Shikimato-3-Fosfato Sintase (EPSPs) e seu inibidor o herbicida glifosato (GPJ). A participação do substrato Shikimato-3- Fosfato (S3P) também foi estudada. A estabilidade e a distribuição de cargas ao longo da superfície da enzima EPSPs foram analisadas para subsidiar o desenvolvimento de um nanobiossensor específico para a detecção do herbicida glifosato através da funcionalização da ponta de um microscópio de força atômica (AFM) com a enzima EPSPs. A parametrização de dois ângulos diedros para o herbicida GPJ foram obtidos através de cálculos mecânico- quânticos ao nível de Hartree Fock, utilizando como conjunto de bases o 6-31G*. Os parâmetros obtidos, relacionados aos coeficientes de Ryckaert-Bellemans, foram implementados no campo de força OPLS-AA, utilizado pelo programa GROMACS. Simulações de dinâmica molecular direcional (SMD), foram realizadas em três possíveis caminhos de retirada do herbicida do sítio ativo, aplicando-se forças harmônicas externas com constante de força equivalente à do cantiléver do AFM, com o objetivo de simular experimentos de detecção do herbicida por microscopia de força atômica. Uma expansão cumulante de segunda ordem da igualdade de Jarzynski foi utilizada para os cálculos do perfil de energia livre ao longo da coordenada de reação e, portanto, para se obter o Potencial de Força Média (PMF) das simulações de SMD. Os três caminhos escolhidos apresentaram diferentes barreiras energéticas relacionadas com o rompimento das interações do inibidor com a enzima. Estas barreiras energéticas calculadas pelas simulações estão na mesma ordem de magnitude das energias livres de ligação para sistemas semelhantes obtidas a partir das constantes de inibição experimentais. / This work aims to developed a molecular model that can describe equilibrium properties, as well as the interacting forces involved in an enzyme-inhibitor system, where the enzyme is the 5-Enolpyruvyl-Shikimate-3-Phosphate Synthase (EPSPs) and its inhibitor is the glyphosate herbicide (GPJ). The participation of the substrate Shikimate-3-Phosphate (S3P) was also studied. The stability and distribution of charges along the surface of the EPSPs enzyme were analyzed for the development of a specific nanobiosensor for glyphosate herbicide detection through the functionalization of the tip of an atomic force microscope (AFM) with the enzyme EPSPs. The parameterization of two dihedral angles for the GPJ herbicide was obtained through mechanical-quantum calculations at the Hartree Fock level using the 6-31G* base set. The parameters obtained, related to the Ryckaert-Bellemans coefficients, were implemented in the OPLS-AA force field, used by the GROMACS program. Steered Molecular Dynamics Simulations (SMD) were performed in three possible unbinding pathways, to remove the herbicide from the active site, applying external harmonic forces with a force constant equivalent to that of the AFM cantilever, in order to simulate experiments to detect the herbicide by atomic force microscopy. A second-order cumulant expansion of the Jarzynski’s equation was used to calculate the free energy profile along the reaction coordinate and to obtain the potential of mean force (PMF) of the SMD simulations. The three paths chosen presented different energetic barriers related to the disruption of inhibitor interactions with the enzyme. These energy barriers calculated by the simulations are in the same order of magnitude as the bond free energies for similar systems obtained from the experimental inhibition constants. / Tese (Doutorado)

O presente trabalho descreve o uso de eletrodos de carbono vítreo como eletrodo-base para modificação com hexacianoferratos de Fe (FeHCF), Co (CoHCF) e Ni (NiHCF) por meio da adsorção destes complexos utilizando a técnica de voltametria cíclica. Os eletrodos quimicamente modificados (EQMs) preparados por este método apresentam características diferenciadas tais como robustez e estabilidade ao longo das medidas eletroquímicas, além de boas propriedades redox. Os EQMs com FeHCF e NiHCF foram avaliados em função da sua atividade eletrocatalítica para a reação de oxidação do Ácido Ascórbico em solução de KCl 0,1 mol L-1 (pH 6,6). O EQM com CoHCF foi avaliado nas mesmas condições dos outros filmes, porém não apresentou uma diminuição no sobrepotencial de oxidação do Ácido Ascórbico. Embora não tenha apresentado uma resposta semelhante aos EQMs com FeHCF e NiHCF, o EQM com CoHCF apresentou a melhor sensibilidade da resposta de corrente de pico quando comparado aos demais. A fim de proporcionar a renovação da superfície do eletrodo foi avaliada a utilização de eletrodos de pasta de carbono modificada com FeHCF, CoHCF ou NiHCF. Os parâmetros de composição da pasta, o eletrólito suporte e o pH foram avaliados utilizando o EPCM com FeHCF por meio da técnica de voltametria cíclica e os parâmetros otimizados foram aplicados no uso dos EPCM com CoHCF e NiHCF. Para avaliar a sensibilidade na deteccção da L-Cisteína, do Glifosato e da N-Acetilcisteína, os EPCMs modificados foram submetidos a estudos com a técnica de voltametria de onda quadrada. Todos os analitos apresentaram resposta eletroquímica em todos os EPCMs, porém as melhores respostas foram para o FeHCF na presença de L-Cisteína, CoHCF na presença de Glifosato e NiHCF na presença de N-Acetilcisteína. Na presença de 1 x 10-8 mol L-1 dos analitos os EPCMs com hexacianoferratos apresentaram um boa resposta eletroquímica, embora não tenha sido possível observar uma linearidade entre o aumento da concentração dos analitos propostos e a corrente de pico. Essa sensibilidade demonstrada pelos EPCMs na presença de moléculas com grupos funcionais, como ácidos carboxílicos, aminoácidos e tióis abre novas perspectivas para a análise direta de compostos de interesse clínico, farmacêutico e amostras de interesse ambiental. / This work describes the use of glassy carbon electrodes as the base for electrode modification with hexacyanoferrates of Fe (FeHCF), Co (CoHCF) and Ni (NiHCF) through adsorption of these complexes using cyclic voltammetry. The chemically modified electrodes (CMEs) prepared by this method have specific features such as robustness and stability along the electrochemical measurements, and good redox properties. The CME with FeHCF and NiHCF were evaluated take into accounting their electrocatalytic activity for oxidation reaction of Ascorbic Acid in KCl 0.1 mol L-1 (pH 6.6). The CME with CoHCF was evaluated under the same conditions, but did not show a decrease on the overpotential for Ascorbic Acid oxidation. Although the CME with CoHCF has not showed the same behavior presented by CMEs with FeHCF and NiHCF, the CME with CoHCF presented the best sensitivity in order to relationship between peak current and concentration of Ascorbic Acid when compared to CMEs with either. In order to provide the renewal of the electrode surface was evaluated the use of carbon paste electrodes (CPEs) modified with FeHCF, CoHCF and NiHCF. The CPE parameters such as composition, supporting electrolyte and pH were evaluated using the CPE with FeHCF by cyclic voltammetry and these optimized parameters were applied for CPEs with CoHCF and NiHCF. The studies about the sensitivity of the detection for L-Cysteine, glyphosate and N-acetylcysteine using the modified CPEs with hexacyanoferrates were performed by square wave voltammetry. All analytes showed electrochemical response at all CPEs, but the best electrochemical responses were obtained for FeHCF in the presence of L-Cysteine, CoHCF in the presence of glyphosate and NiHCF in the presence of N-acetylcysteine. The CPEs with hexacyanoferrates in the presence of 1 x 10-8 mol L-1 of L-Cysteine, glyphosate or N-acetylcysteine had showed a good electrochemical response, although were not possible to observe a linear correlation between the concentration of analytes and peak current. This sensitivity demonstrated by CPEs with hexacyanoferrates electrodes in presence of specific functional groups such as carboxylic acids, amino acids and thiols opens new perspectives for the direct analysis of compounds with clinical, pharmaceutical and environmental samples.

La qualité de l'eau du lac Saint-Pierre (LSP), le plus grand lac fluvial du fleuve Saint-Laurent, est notamment compromise par le déversement d’une mixture composée de métaux et de pesticides provenant des rejets des industries, des effluents municipaux et de l’exploitation des terres agricoles dans son bassin versant. Cette contamination est d'autant plus importante dans les zones du lac caractérisées par une végétation dense favorisant la rétention et la sédimentation de la matière en suspension. Or, ces herbiers aquatiques, qui occupent de vastes étendues du LSP, servent de frayère pour plusieurs poissons, dont la perchaude (Perca flavescens). Cette espèce est donc particulièrement affectée par la dégradation des habitats aquatiques du LSP. À la suite d’un déclin important de ses populations depuis la fin des années 1990 en raison de son important intérêt commercial et sportif, un moratoire de cinq ans sur la pêche de la perchaude a été imposé en 2012 et reconduit jusqu’en 2022, puisque l’incapacité de rétablissement des populations de perchaudes semble persister. La présente étude vise à évaluer l'hypothèse que cette incapacité de rétablissement, qui se reflète par un recrutement déficient, soit en partie attribuable à l'impact de la contamination chimique sur la reproduction, soit par des effets toxiques potentiels sur les femelles ovigères, sur les œufs ou sur les jeunes larves pendant les premiers mois. Dans un premier temps, nous avons évalué l’hypothèse que le glyphosate, herbicide à large spectre et ingrédient actif de la formulation Roundup®, ait un impact indirect sur les jeunes perchaudes en décimant les communautés de biofilms périphytiques, source d’alimentation des invertébrés dont se nourrissent les jeunes larves. Cette suite d’effets contribuerait à l’accroissement de la mortalité hivernale des jeunes de l’année, due à une insuffisance de ressources énergétiques. Or, nos résultats démontrent que peu importe l’âge, et par le fait même l’épaisseur des biofilms, le glyphosate, en concentrations environnementales réalistes, ne semble pas impacter négativement la composition des communautés ou le métabolisme de la chlorophylle des biofilms. Seul l’âge (2 mois, 1 an, 20 ans) de ces derniers semblait en effet influencer la composition taxonomique des communautés. Nous avons cependant observé une augmentation de l’abondance relative d’Anabaena, un taxon de cyanobactérie toxique qui possède une forme résistante rare de l’enzyme EPSPS, cible principale du mode d’action du glyphosate. Cette étude contribue à l’avancement des connaissances sur les effets de l’herbicide le plus utilisé à l’échelle mondiale, actuellement au cœur de préoccupations d’intérêt international. Nous avons également évalué le potentiel de toxicité associé au transfert maternel du mercure et du sélénium chez la perchaude à l’aide de techniques de fractionnement subcellulaire. Le mercure est un contaminant d’intérêt en raison de son omniprésence dans l’environnement ainsi que de ses effets néfastes sur la reproduction des poissons à de très faibles concentrations. Il a également été démontré que des ratios molaires Se:Hg supérieurs à 1 atténuaient les effet néfastes du mercure. Nos résultats démontrent une évidence de transfert maternel de la femelle à ses œufs, mais également aux mitochondries gonadiques, principales composantes sensibles de la cellule. Le transfert maternel représentant la source d’exposition aux contaminants la plus importante pour les embryons, nos observations pourraient contribuer à expliquer le recrutement déficient des jeunes perchaudes au LSP. Nous avons également mesuré des ratios molaires Se:Hg systématiquement supérieurs à 1 dans les différentes fractions subcellulaires hépatiques et gonadiques, résultats novateurs qui laissent sous-entendre un effet protecteur du Se. Puisque nous avons confirmé l’occurrence d’un transfert maternel du mercure, l’étape logique subséquente était d’évaluer la bioaccumulation de ce contaminant au sein des différents stades ontogéniques du cycle de vie de la perchaude. Les stades embryo-larvaires et juvéniles précoces sont en effet des phases particulièrement sensibles aux contaminants organiques et inorganiques. Nos résultats démontrent que les concentrations de MeHg décroissent suivant un patron ontogénique, avec les plus hautes concentrations mesurées chez les juvéniles et les plus basses dans les masses d’œufs. Nous avons également démontré que presque 100% du mercure était présent sous forme de MeHg, forme toxique et bioamplifiable, chez les larves et les juvéniles. Les ratios molaires Se:Hg étaient quant à eux systématiquement supérieurs à 1, résultats comblant d’importantes lacunes au niveau des effets antagonistes entre le mercure et le sélénium chez les poissons. Les résultats découlant des présents travaux de recherche ont un impact important sur la science de l’écotoxicologie en raison de leur caractère novateur. Tout d’abord, nous avons contribué à l’avancement des connaissances sur l’impact de concentrations environnementales de glyphosate sur des biofilms d’âge très contrasté. Ensuite, nous avons, pour la première fois, utilisé des outils de fractionnement subcellulaire afin d’évaluer le potentiel de toxicité lié au transfert maternel du mercure. Enfin, nous rapportons les premières données liées à la bioaccumulation simultanée du mercure et du sélénium aux stades de vie clés du développement de la perchaude. La présente thèse s’avère ainsi nécessaire afin de contribuer au progrès du savoir sur le devenir de certains contaminants d’intérêt au sein des écosystèmes aquatiques. / Lake Saint Pierre (LSP) is the largest fluvial lake in the Saint Lawrence River. Water quality in LSP is heavily affected by inputs of nutrients and chemical pollution from tributaries which drain agricultural watersheds, from municipal effluents and from industrial discharges. This contamination is amplified in areas of LSP with dense vegetation because aquatic plants promote the retention and sedimentation of dissolved and particulate matter. Several fish species, including Yellow Perch (Perca flavescens), use these aquatic vegetation beds as their spawning grounds and are therefore particularly affected by the contamination of aquatic habitats in LSP. This study therefore tests the hypothesis that chemical contamination in Yellow Perch (YP) during their early life stages can help explain this species’ lack of resilience despite the implementation of a fishing moratorium in 2012. This moratorium was extended until 2022 since populations are still undergoing a recruitment failure and a decline in juvenile abundance populations. The phosphonate herbicide glyphosate, which is the active ingredient in Roundup®, is currently the most widely used herbicide in the world. Glyphosate-based herbicides are sprayed on food and feed crops during cultivation and are thus subject to leaching to streams and rivers. In aquatic ecosystems, periphytic biofilms, or periphyton, are important primary producers and are often the first trophic level to be in contact with runoff waters. Thus, a trophic cascade could occur if these biofilms are negatively impacted by glyphosate, potentially leading to larval fish mortality due to resource limitation. Results showed that submersion period (2 months, 1 year, 20 years) was the only significant contributor to community structure. However, the glyphosate-resistant Cyanobacteria Anabaena was found to be favoured by the use of glyphosate. This freshwater Cyanobacteria commonly forms toxic blooms, raising concern regarding the use of glyphosate. For all colonization stages, and therefore different thicknesses, chlorophyll a did not show an unequivocal decline over time. This study therefore provides an interesting snapshot of the biological processes related to periphytic biofilms’ exposure to environmental concentrations of glyphosate. As this herbicide is currently of international concern, it is imperative to contribute to the advancement of knowledge about its effects. Mercury (Hg) is a trace element of particular concern since it is ubiquitous in the environment and because its methylated form (MeHg) readily bioaccumulates and biomagnifies in food webs. This latter process leads to elevated Hg concentrations in fish and thus induces toxicity. Maternal transfer of bioaccumulated contaminants to offspring is a suggested mechanism of impaired reproductive success in fish. We therefore assessed the toxicity potential of Hg during maternal transfer in YP from LSP using a sub-cellular partitioning approach. Results showed a strong relationship between Hg bioaccumulation in the liver and Hg concentrations in gonadal mitochondria, which corroborates the potential toxicity of maternal transfer. As selenium is a well-studied Hg antagonist, we also measured the Se:Hg molar ratios in all subcellular fractions. We found that these ratios were systematically above 1, which is the suggested threshold for Hg toxicity alleviation through sequestration by Se. Since early developmental stages in aquatic biota are particularly sensitive to Hg, and after confirming the evidence of maternal transfer, we subsequently addressed Hg bioaccumulation in all parts of YP life cycle. This study is the first of its kind to follow Hg and Se during YP ontogenetic development, from the gravid female to the juvenile. Results show that MeHg follow an ontogenetic pattern, with concentrations decreasing from the juveniles to egg masses. We also found that nearly 100% of THg was measured as the toxic form MeHg in larvae and juveniles. Lastly, Se:Hg molar ratios were systematically above 1, suggesting a potentially protective effect of Se on Hg bioaccumulation. This study will thus provide much needed information on the changes in bioaccumulation patterns during the most sensitive life cycle stages of this declining fish population. Results issued from the present research have significant impact when it comes to the advancement of knowledge in Ecotoxicology due to their novel characteristics. First, we have contributed to the advancement of knowledge on the effects of environmental concentrations of glyphosate on highly age-contrasted biofilms. Also, it is the first time that subcellular partitioning techniques are used in order to assess the toxicity potential of mercury during maternal transfer. Finally, we provide the very first results on the simultaneous bioaccumulation of mercury and selenium in key life stages of YP development. Therefore, this thesis is of particular interest when aiming to assess the fate of certain contaminants of interest within aquatic ecosystems.

Holistic environmental policies, which emerged from a mere combination of technical activities in waste management some 40 years ago, constitute the most advanced level of environmental policies. These approaches to environmental policy, among them the policies in integrated waste management, attempt to guide economic agents to an environment-friendly behaviour. Nevertheless, current holistic policies in waste management, including policies on one-way drinks containers and waste electrical and electronic equipment, and implementations of extended producer responsibility with further applications to waste electrical and electronic equipment, reveal more or less severe deficiencies – despite some positive examples. This article relates these policy failures, which are not necessarily the result of an insufficient compliance with the regulations, to missing constitutive elements of what is going to be called an ‘integrated environmental policy’. This article therefore investigates – mostly from a practical point of view – constitutive elements, which are necessary for a holistic policy to serve as a well-functioning allocation mechanism. As these constitutive elements result from a careful ‘integration’ of the environmental commodities into the economic allocation problems, we refer to these policies as ‘integrated environmental policies’. The article also discusses and illustrates the main steps of designing such a policy – for waste electrical and electronic equipment and a (possible) ban of Glyphosat in agriculture. As these policies are dependent on economic and political stability with environmental awareness sufficiently developed, the article addresses mostly waste management policies in highly industrialised countries.