Orientadores: Susanne Rath, Sonia Claudia do Nascimento de Queiroz / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-27T01:49:14Z (GMT). No. of bitstreams: 1 Porto_RafaelSilveira_M.pdf: 1983268 bytes, checksum: f89fcb853911e559df9889c577be5a8e (MD5) Previous issue date: 2015 / Resumo: O Brasil é um dos maiores produtores de frutas do mundo, no entanto, estima-se que doenças pós-colheita possam gerar perdas de até 50% em sua produção. A forma mais comum de tratamento para essas doenças envolve a aplicação de fungicidas sintéticos. Contudo, nos últimos anos, a demanda por tratamentos alternativos tem crescido, com destaque para o uso de biopesticidas, produtos desenvolvidos a partir de plantas, microrganismos e insetos. Este trabalho teve como objetivo investigar a presença dos compostos bioativos (4Z)-lachnophyllum lactona, (4Z,8Z)-matricaria lactona e (2Z,8Z)-matricaria ester nos espécimes brasileiros da planta Conyza canadensis, bem como avaliar a atividade antifúngica dessas substâncias isoladas contra diversos fungos associados a doenças pós-colheita de frutas. Por cromatografia flash preparativa foi possível isolar a (4Z)-lachnophyllum lactona e a (4Z,8Z)-matricaria lactona a partir de extratos da planta obtidos com diclorometano. Os compostos foram caracterizados por GC-MS/MS, NMR 1H e 13C, 1H-1H COSY e 1H-13C HSQC. Foram realizados ensaios de difusão em disco com 10 fungos filamentosos causadores de doenças pós-colheita em frutas. Os fungos Aspergillus niger, Cladosporium spp. e Penicillium digitatum se mostraram susceptíveis ao tratamento e, para eles, a concentração mínima inibitória dos compostos variou de 32 a 64 µg mL-1. Também foi desenvolvido um método de extração empregando água quente pressurizada, no qual foram otimizados os parâmetros de temperatura (100 °C), tempo de ciclo (1 min) e número de ciclos (quatro). Com essa técnica foi possível obter um rendimento de 1,46 mg g-1 e 0,24 mg g-1 para a (4Z)-lachnophyllum lactona e a (4Z,8Z)-matricaria lactona, respectivamente. O extrato aquoso da Conyza canadensis pode ser aplicado diretamente nos frutos com a vantagem de não conter resíduos de solventes orgânicos tóxicos / Abstract: razil is one of the largest fruit producers in the world. Nevertheless, it is estimated that postharvest diseases can lead to losses of up to 50% in its production. The most common treatment for these diseases involves the application of synthetic fungicides. Nonetheless, in recent years, the demand for alternative treatments has increased, especially for the use of biopesticides, products developed from plants, microorganisms and insects. This study aimed to investigate the presence of the bioactive compounds (4Z)-lachnophyllum lactone, (4Z,8Z)-matricaria lactone and (2Z,8Z)-matricaria ester in Brazilian specimens of the weed Conyza canadensis, as well as to evaluate the antifungal activity of these isolated substances against several fungi associated with postharvest diseases of fruits. With the use of preparative flash chromatography it was possible to isolate (4Z)-lachnophyllum lactone and (4Z,8Z)-matricaria lactone from plant extracts obtained with dichloromethane. The compounds were characterized by GC-MS/MS, NMR 1H e 13C, 1H-1H COSY and 1H-13C HSQC. Disk diffusion assays were performed in order to investigate the activity of the isolated compounds against 10 filamentous fungi regarded as common postharvest pathogens of fruits. Aspergillus niger, Cladosporium spp. and Penicillium digitatum proved susceptible to the treatment and, for them, the minimum inhibitory concentration of the compounds varied from 32 to 64 µg mL-1. An extraction method using pressurized hot water was also developed, in which the parameters of temperature (100 ° C), cycle time (1 min) and number of cycles (four) were optimized. By using this technique, it was possible to obtain a yield of 1.46 mg g-1 and 0.24 mg g-1 for the (4Z)-lachnophyllum lactone and (4Z,8Z)-matricaria lactone, respectively. The aqueous extract of Conyza canadensis can be applied directly on fruits with the advantage of not containing residues of toxic organic solvents / Mestrado / Quimica Analitica / Mestre em Química

Agricultural crops are a food subsidy for wild ungulates that can bring animals into close proximity, facilitating disease spread. We used stable isotope analysis to reconstruct the diets of elk and white-tailed deer in three areas in the Canadian prairies and calculated their isotopic niche breadth. Isotopic niche breadth was greater for deer than elk, indicating that deer are a generalist species composed of individual specialists with varied diets, while elk are individual generalists. White-tailed deer niche breadth decreased with increasing consumption of agricultural foods in early fall, but not on an annual timescale. Elk niche breadth did not change with consumption of agricultural foods. Agricultural sources were 40-80% of the diets of both species in all areas. Agricultural feeding may increase the risk of disease in elk and deer not only by increasing their apparent density at feeding sites, but also by subsidizing increases to their overall density.

The Hutchinsonian niche is the n dimensional hyper volume that allows for the persistence of a species. Castor canadensis, a large semi-aquatic rodent, is an ecosystem engineer and often a keystone species for many ecosystems. I examined the effect of multiple spatial scales on hierarchical habitat selection byC. canadensis using presence-only modeling techniques. I also determined individual trophic niche specialization in C. canadensis utilizing stable isotope analysis. I concluded that C. canadensis displayed scale independent habitat selection when comparing landscape and fine spatial scales. Individual trophic niche specialization occurred in colonies of the same resource availability. Also, individual trophic niches varied substantially between wetlands. These results have implications for the management of “generalist” species because populations can be composed of specialized individuals. Studies of niche across spatial and organizational scales are required for successful conservation and management strategies.

The hemlock woolly adelgid (HWA), Adelges tsugae Annand, (Hemiptera: Adelgidae) is an invasive species in the eastern United States and Canada. This scale-like insect feeds on the xylem ray parenchyma cells of eastern hemlock, Tsuga canadensis (L.) Carrière, (Pinales: Pinaceae) and Carolina hemlock, Tsuga caroliniana Englem., resulting in multiple physical and physiological symptoms, all of which commonly leads to tree mortality within 4 to 10 years. Currently, HWA has spread throughout most of the eastern hemlock range and all of the range of Carolina hemlock and threatens the forest ecosystems they serve. In its introduced range, there are no natural enemies specialized to feed on HWA and suppress HWA populations below damaging levels. In the urban environment and at some public locations, the use of chemicals, such as imidacloprid, have been used to temporarily protect hemlocks, however; this does not provide a permanent solution. In the 1990's a classical biological control program for HWA was created with the goal to suppress HWA populations below damaging levels to reduce tree mortality, and to reduce the negative effects associated with the loss of either hemlock species. Presently, four predators have become the main focus of this program: Laricobius nigrinus Fender, Laricobius osakensis Shiyake and Montgomery (Coleoptera: Derodontidae), Leucotaraxis argenticollis (Zetterstedt), and Leucotaraxis piniperda (Malloch) (Diptera: Chamaemyiidae). Throughout the eastern US, La. nigrinus has established populations and expanded its distribution beyond where it has been released. Although it has successfully established and continues to spread, its effect on eastern hemlock health has been unknown. A 1-year study, carried out in 2021, revealed that La. nigrinus predation on the sistens generation not only reduced the sistens generation, but also had a similar net photosynthetic rate to the negative control on 1-2 year old shoot growth. In addition, significantly more shoots were produced for treatment branches with the highest La. nigrinus density. This also led to a significantly reduced aestivating sistens density compared to the no predator treatment. However, predation was insufficient to reduce HWA populations below the suggested damaging threshold of eastern hemlock (< 4 HWA/cm), suggesting that additional predators would be needed. Recently, the western strain of Le. argenticollis had been released in the eastern US. However, its phenology in the eastern US was unknown. A 2-year study, from 2021-2023, revealed that Le. argenticollis eggs and larvae were present when both generations of HWA adults with eggs and HWA nymphs were present. In addition, Le. argenticollis was capable of completing development in NY and VA, showing promise that this species could establish populations in the eastern US. Timing the adult fly releases into cages at the time when sistens were producing eggs resulted in Le. argenticollis phenology to synchronize well with HWA phenology at both locations, thus providing additional support for its release in the eastern US. Another 2-year study conducted in 2021-2022, at five locations (MD, NC, PA, TN, and VA), sought to determine how the predation of La. nigrinus and Le. piniperda, alone and together, would impact the sistens and progrediens generations, as well as how their predation would affect the aestivating sistens density and new shoot growth production. From this study, La. osakensis was found as the dominant Laricobius species present at the TN site, and similarly to La. nigrinus, reduced intact ovisac density in. This provided a small preview in La. osakensis capabilities and how this predator may be impacting HWA populations at locations where it has established. In order to assess Le. piniperda's impact on the progrediens generation, mesh cages were applied to treatment branches at each site. The study found that Le. piniperda could reproduce at all field sites and that the percentage of disturbed progrediens was significantly higher for treatments with Le. piniperda present at all field sites, except at PA in 2022. However, the affect of both of these predators on the aestivating sistens density and eastern hemlock health was inconclusive. In addition, the percentage of new shoot growth was highly variable among treatments at all field sites, and there was no treatment effect on the mean length of new shoot growth produced at all field sites. The results of this study suggests that in order to fully understand the affects of these predators on HWA populations and on hemlock tree health, longer term studies should be conducted. However, this study does provide some insight on the capabilities of these predators in the short term and supports the continued release and redistribution of these predators. / Doctor of Philosophy / The hemlock woolly adelgid (HWA), is an invasive species in the eastern United States and in Canada. This small scale-like insect was introduced likely on nursery stock material from southern Japan and was first discovered in the eastern US in 1951 in Richmond, Virginia. Two generations of HWA occur each year known as the sistens and progrediens; both reproduce without males. This insect feeds on eastern hemlock and Carolina hemlock, and since its discovery has spread through most of their range. While this insect feeds, it weakens the tree, eventually causing tree death. In eastern North America, eastern hemlock is one of the major tree species in the forest and provides a unique habitat for various mammals, birds, fish, and invertebrate species. To protect this habitat, multiple management strategies have been developed. The two main management strategies that are in use are the use of pesticides and the release of predators of HWA. Pesticides have been useful in the urban environment and in parks and forests, but they only last for a short time and can lead to negative affects to the environment. Since the 1990's, the release of insect predators originally found in the native range of HWA, has occurred with the goal of predator establishment, to lower HWA populations, and to lower hemlock tree death in the eastern US. Four small insect predators have been the main focus of this program: Laricobius nigrinus, Laricobius osakensis, Leucotaraxis argenticollis, and Leucotaraxis piniperda. So far La. nigrinus and La. osakensis have populations at multiple locations throughout the eastern US, and a low number of sites have been recently found for Le. argenticollis and Le. piniperda. Past studies have indicated that La. nigrinus has a significant impact to the sistens generation of HWA, but its affect to eastern hemlock health has been unclear. A 1-yr study conducted in VA in 2021, released La. nigrinus adults into mesh cages, with two different population sizes of beetles, to determine if La. nigrinus predation on HWA populations lead to an indirect effect on eastern hemlock health at the branch level. Results confirmed that La. nigrinus had a significant impact to the sistens generation of HWA. Photosynthesis of branches with La. nigrinus, in June, was similar to the photosynthesis of the negative control branches, which started with a lower number of sistens present on the branch at the beginning of the study. Suggesting that branches with La. nigrinus may be healthier compared to branches without La. nigrinus. Laricobius nigrinus predation also significantly reduced the number of aestivating sistens on new shoot growth, but not below damaging levels (< 4 HWA/cm). This suggested that additional predators may be needed to target the progrediens generation of HWA so that HWA populations do not return to damaging levels after La. nigrinus predation. To learn more about the potential for Le. argenticollis to survive and reproduce in the eastern US, a 2-yr study was conducted in 2021 – 2023, to determine when different life stages of Le. argenticollis could occur in NY and VA. Results revealed that Le. argenticollis was able to reproduce in the mesh cages and complete its life cycle. In addition, Le. argenticollis eggs and larvae were present at the time when HWA adults were producing eggs and when HWA nymphs were present for both generations of HWA. These results provide insight on Le. argenticollis life cycle and shows that Le. argenticollis has a good chance in surviving throughout the eastern US. A 2-yr study at five locations in the eastern US (MD, NC, PA, TN, and VA), tried to combine two predators, La. nigrinus and Le. piniperda, to determine if their combined impact would be sufficient to reduce HWA populations and affect eastern hemlock tree health. Three assessments were conducted each year to determine the impacts of La. nigrinus and Le. piniperda on HWA populations and on eastern hemlock shoot growth production as a representation of tree health. Results confirmed that La. nigrinus significantly reduced the sistens generation at all locations, except at TN. At the TN site, La. osakensis was found to be the main predator of the sistens generation and had a similar affect on the sistens generation as La. nigrinus. Leucotaraxis piniperda was able to reproduce at all locations and also had a significant affect on progrediens ovisacs at four of the locations in 2021 (MD, NC, PA, and VA) and at three of the locations in 2022 (MD, NC, and VA). However, the results on the number of aestivating sistens on new shoot growth and eastern hemlock health were inconclusive. Therefore, it is unclear as to how the presence of both of these predators' affect eastern hemlock health. A long term study may be needed in order to understand the relationship between these predators and their indirect affect to eastern hemlock health.

The biosynthesis of hydrastine and of berberine was investigated by feeding radioactive tyrosine and dopamine to Hydrastis canadensis L. It was shown that both hydrastine and berberine are specifically derived from two molecules of tyrosine, but that the ratio of incorporation of the two molecules of radiotyrosine was not unity, but varied with time. Dopamine was also incorporated into hydrastine without randomization, but only one molecule was utilized. The results, which are consistent with Robinson’s hypothesis of biogenesis, throw light on some of the individual steps of the biosynthetic path by which hydrastine and berberine are derived. / Thesis / Doctor of Science (PhD)

Prey-mediated effects of imidacloprid were evaluated for Laricobius nigrinus Fender and Sasajiscymnus tsugae Sasaji and McClure after feeding on hemlock woolly adelgid (HWA), Adelges tsugae Annand (Hemiptera: Adelgidae). Two methods were evaluated for detecting imidacloprid in hemlock tissues: a commercially available enzyme linked immunoassay (ELISA) kit and a high performance thin-layer chromatography technique for detecting and quantifying imidacloprid residues in hemlock wood and needle tissues. ELISA is advantageous because of its cost, sensitivity, and ease of use. However, matrix effects in the form of false positives and overestimated imidacloprid concentrations were evident in hemlock wood and needle tissue extracts. Matrix effects could be reduced by dilution with water, effectively raising the lower detection range of the kit from 0.2 to 200 ppb. High performance thin-layer chromatography was accurate, quick, easy to use, and matrix effects were not evident. However, the technique is sensitive in the lower ppm range and tissue samples from field-treated hemlocks are often in the ppb range, making this technique less desirable than more sensitive analytical methods. Lethal and sublethal effects on both predators were evident after eastern hemlock branches infested with HWA were spiked with imidacloprid in the laboratory. HWA mortality increased with dosage and time, and its 30 d LC50 was determined to be 242 ppb. Both predator species exhibited reduced survivorship and fitness parameters after feeding on HWA from the treated branches. In a topical application bio-assay, 6 d imidacloprid LD50 values for L. nigrinus and S. tsugae were 2.43 and 1.82 µg/g, respectively. Imidacloprid and its major metabolites in hemlock tissues were analyzed by liquid chromatography-tandem mass spectrometry. Imidacloprid recovery from beetle cadavers was correlated with beetle mortality from feeding on treated hemlock branches. Olefin was the primary imidacloprid metabolite recovered from hemlock wood tissues. When predators fed on HWA from field-treated trees, impacts on survivorship and fitness were variable. In 2005, significantly higher proportions of both species of beetles were affected by feeding on control branches compared with treated branches. In 2006, beetles feeding on HWA from some of the trees treated in the field exhibited longer fliptimes compared with beetles feeding on controls, although beetle mortality was not significant among treatements. In the field, imidacloprid controlled HWA populations 1-3 years post-treatment. Hemlock health improved in the highest dosage group, with significantly greater lengths of new shoots compared with shoots from control trees. Eastern hemlock trees primarily metabolized imidacloprid into the olefin metabolite, which can have increased insecticidal toxicity compared with imidacloprid. Imidacloprid was detected in beetle cadavers after feeding on HWA from treated branches, suggesting that prey-mediated impacts of systemic imidacloprid are possible on nontarget predators. However, because of HWA's sensitivity to imidacloprid, in field situations predators are more likely to be affected by reduced adelgid density and quality. / Ph. D.

Tyrimo objektas – kanadinės rykštenės lapų bei žiedų ekstraktai. Tyrimo metu kokybiniam ir kiekybiniam flavonoidų nustatymui naudojamas efektyviosios skysčių chromatografijos metodas. Darbo tikslas – nustatyti flavonoidų kokybinę ir kiekybinę sudėtį kanadinės rykštenės (Solidago canadensis L.) augalinėje žaliavoje naudojant ESC metodą. Atlikta mokslinių literatūros šaltinių analizė, pagrindžianti kanadinės rykštenės (Solidago canadensis L.) gydomąsias savybes, flavonoidų išskyrimo bei nustatymo metodikas , taip pat taikomo metodo optimizavimo ir validacijos kriterijus. Optimizuotas ECS metodas flavonoidų vertinimui, naudojant turimą laboratorinę įrangą. Pasirinkta 150×4,6 mm, 3 µm YMC kolonėlė, kurios temperatūra 25°C. Injekcijos tūris – 10µl. Kiekybiniam nustatymui pasirinktas gradavimo grafiko metodas, taikant tiesines kalibracines kreives. Validacijos eksperimentų metu buvo pagrįstas metodikos pakartojamumas, tiesiškumas, aptikimo ir nustatymo ribos. Validuota metodika pritaikyta kanadinės rykštenės (Solidago canadensis L.) ekstrakto tyrimui, nustatyta, kad kanadinės rykštenės lapų ekstrakte flavonoidų vidutiniškai yra 23,0 mg/g, iš jų vidutiniškai rutino nustatyta 18,99 mg/g, izokvercitrino – 0,44 mg/g, hiperozido 0,19 mg/g, kvercitrino 3,38 mg/g. Kanadinės rykštenės žiedų ekstraktuose flavonoidų nustatyta vidutiniškai 23,27 mg/g, iš jų rutino nustatyta 19,70 mg/g, izokvercitrino – 0,68 mg/g, hiperozido – 0,47 mg/g, o kvercitrino – 2,42 mg/g. / The object of the research – extracts of Canadian goldenrod leaves and flowers. The high performance liquid chromatography method was used for the quantitative determination of flavonoids in the research. The aim of work was to determine the qualitative and quantitative composition of flavonoids in the plant raw material of Canadian goldenrod (Solidago canadensis L.) using HPLC method. Analysis of scientific literature was performed to support the medicinal properties of the Canadian goldenrod (Solidago canadensis L.), methods for extraction and detection of flavonoids, as well as the validation criteria of the applied optimization method. The HPLC method for determining the quantity of flavonoids was optimized using the available laboratory equipment. YMC column of 150×4.6 mm, 3 µl, was chosen with a temperature of 25°C. The injection volume – 10 µl. The graph calibration method was selected for using linear calibration curves. Validation experiments justified the method repeatability, linearity and the limits. The validated methodology was applied for the research of Canadian goldenrod (Solidago canadensis L.) extract, and it was determined that the leaf extract of Canadian goldenrod contains an average of 23.0 mg/g flavonoids, of which, the average amount of rutin found was 18.99 mg/g, isoquercitrin – 0.44 mg/g, hyperoside 0.19 mg/g, quercitrin 3.38 mg/g. The average flavonoid content in the flower extract of Canada goldenrod was 23.27 mg/g, of them rutin 19.70 mg/g... [to full text]

Ce mémoire examine l’ethos d’historien de François Du Creux dans ses Historiæ canadensis publiées en 1664. Cette œuvre relativement peu connue, qui consiste, pour une bonne part, en une réécriture latine des Relations jésuites de Nouvelle-France, pose des problèmes de statut et de finalité découlant notamment des transformations que connaissait l’écriture de l’histoire à l’époque de Du Creux. L’étude de l’ethos est ici envisagée comme un moyen d’aborder les grands enjeux de l’œuvre en précisant le projet auctorial lié au statut de l’historien. À partir des occurrences de la première personne dans les pièces liminaires et dans le corps du texte, nous dégageons d’abord les trois statuts énonciatifs de la figure auctoriale, en tant que Français, Jésuite et historien. Ces statuts évoquent des questions religieuses et politiques, souvent polémiques, qui sont associées à la Compagnie de Jésus au xviie siècle. L’auteur les convoque pour établir son autorité en se référant à sa tâche d’historien, en défendant ses positions sur les questions qui concernent son ordre et en faisant valoir sa familiarité avec les acteurs et les faits historiques. Nous montrons enfin comment les figures adoptées par l’auteur orientent la lecture des événements racontés et affinent la compréhension de la visée de l’œuvre. / This thesis examines the ethos François Du Creux as a historian in his Historiæ canadensis published in 1664. This relatively unknown work, which mostly consists of a rewriting of the New France Jesuits’ Relations, presents problems of status and purpose largely derived from the transformations that were characteristic of the writing of history in Du Creux’s time. The study of the ethos is used here as a way to address the key issues of the book by clarifying the authorial work associated with the status of the historian. First, we have drawn from occurrences of verbs in the first person in the introductory pieces and the main text three descriptive statuses of the authorial figure: Frenchman, Jesuit, and historian. Theses statuses are linked to religious and politic matters, often controversial, that were associated with the Society of Jesus during the 17th century. Du Creux invokes them to establish his authority by citing his historian function, by defending his positions on matters that concern his order, and by asserting his familiarity with the historical players and facts. Lastly, we demonstrate how the figures used by Du Creux guide the reader’s understanding of the events that are described and deepen the comprehension of the work’s aim.

Methylmercury (MeHg) is the most toxic form of Hg to wildlife, partly because of its ability to biomagnify through the food chain. Previous studies have shown that river otter (Lutra canadensis) is particularly susceptible to increased exposure and body burdens of MeHg due to the piscivorous nature of its diet. The objectives of this study were to attempt to establish a relationship between MeHg concentrations in otter tissues and DNA fragmentation using single cell electrophoresis (comet assay) as well as evaluate the effectiveness of the comet assay as a tool in genotoxicity assessment in field samples. Results of preliminary experiments indicate that both time and storage temperature markedly influence the rate of spontaneous DNA degradation. Increased numbers of freeze/thaw cycling also appear to have a negative effect on DNA integrity. In addition, a cell culture experiment was set up to measure the effects of differing levels of MeHg on a neuroblastoma cell line.

No description available.