This diploma thesis was focused on study of low-molecular antioxidants present in various kinds of fruits, namely berries. In theoretical part detailed characterization of antioxidants studied in presented work and their biological effects are introduced. Further, principals of processing and storage of specified species of fruits are discussed. Changes observed during storage of specific fruits are described too. In practical part changes in content of selected antioxidants in berries were followed. As studied material strawberries, raspberries, blueberries, red and black currants were used. All kind of these fruits was stored in fridge for six months using several different kind of processing. Berries were stored as whole fruits, whole osmotically processed fruits by sucrose, raw pulp and sweetened pulp. At first, starting values of selected antioxidant parameters were measured in fresh fruits (ascorbate, total antioxidant activity, total phenolics, catechins and individual flavonoids). After freezing the same parameters were measured regularly in two month periods. As the most suitable method of fruit processing, with regard to antioxidant content, storage of the whole fruits or pulp can be recommended. Content of ascorbate decreased in all forms and species of the processed fruits during all the time of storage. Oppositely, the content of other studied antioxidants exhibited fluctuations or increasing trend. Sensory analysis was proved for the whole fruits and also for whole sweetened fruits.

<p>  </p> <p>Spotted-wing Drosophila (SWD), <em>Drosophila suzukii</em> (Matsumura), is an economically-important pest of small fruits worldwide. Currently, timing of management is based on morphological identification of adult flies captured in baited monitoring traps; however, distinguishing SWD from other native drosophilids in traps is a time-consuming process that requires magnification. And a degree-day model that could help small fruit growers understand and predict the seasonal activity of this pest has not been developed for Indiana. Due to the low tolerance for maggots in fruit market, most small fruit growers rely on intensive, insecticide applications on a calendar-based schedule without guidance on the activity levels of SWD. </p> <p>A total of 6,051 SWD adults were monitored weekly using commercial Scentry traps at three highbush blueberry orchards during May to August. I applied the published SWD developmental thresholds of 7.2 °C (lower) and 31.5 °C (upper) and the single-sine method to calculate accumulated degree days in the year of 2021 and 2022. A predictive model from two years of data at three locations exhibited an S-shaped curve, with 5%, 25%, and 50% of adults detected at ~907, 1,293, and 1,523 CDD, respectively. By examining infestations in three varieties, ‘Bluecrop’, ‘Blueray’, and ‘Elliot’, I found that blueberry infestation rate increased as the trap captures increased. The use of early-ripening highbush blueberry varieties can reduce infestation and regardless of variety, as berries became softer, the number of SWD egg scars in berries increased.</p> <p>DNA-based diagnostic methods, like loop-mediated isothermal amplification (LAMP), have the potential to improve SWD detection by replacing morphological identification with DNA-based identification. Positive results of the LAMP assay are based on a visible color change from pink to yellow when focal DNA is present. I tested the reliability of LAMP results using SWD DNA and then evaluated the sensitivity of LAMP in discriminating between SWD and two native drosophilids common captured in monitoring traps in Indiana, <em>Drosophila affinis</em> and <em>D. simulans</em>. I found the LAMP assay can quickly and accurately identify SWD with as little as 0.1 ng/μl of DNA. Following optimization, the assay also suggested success in discriminating between SWD and these two native species: it only requires an individual fly, DNA extraction is not necessary. </p> <p>By better predicting seasonal SWD activity and optimizing DNA-based diagnostics for this pest, this study can help improve the timely detection of SWD and the management in small fruit systems. </p>

No description available.

Made available in DSpace on 2017-07-25T19:31:00Z (GMT). No. of bitstreams: 1 Kamila Santana.pdf: 1363071 bytes, checksum: 4f5da813f22ced41d3b26851a927779f (MD5) Previous issue date: 2016-06-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The blueberry is an agricultural crops recently introduced in Brazil and lots of researches are turning to develop and adapt it to the country. Due to its low amount of root hair, it forms associations with microorganisms to expand its zone of nutrient uptake, being the nitrogen the most required element. At this work it was observed the interaction of this cultivation with the inoculum Azospirilum brasilense and Trichodema asperellum into different substrates or doses of nitrogen fertilization. The experiment was performed with cv. Climax and the plants conditioned in pots in greenhouse, under trial design of randomized blocks, factorial 3x3. The considered factors were inoculation (A. brasilense, T. asperellum or without inoculation) and formulation of the substrates (5; 20 e 35% of organic matter – hardened cattle manure) or, at the second experiment, inoculation (A. brasilense, T. asperellum or without inoculation) and doses of nitrogen (0, 10 and 15 g f N plant-1 – provided as urea). The larger quantity of organic matter provided the highest averages relating to physiological (transpiration, water use efficiency and carboxylation efficiency) and growth variables (foliar limb area, total foliar area and rate of relative growth), although, for some of them, it has not differed from plants grown with 20% of organic matter. With 30% of O.M., the inoculation with T. asperellum showed superior averages in CO2 assimilation (A), carboxylation efficiency (A/Ci) and rate of relative growth (RRG). The nitrogen doses positively affected A/Ci, clorophyll amounts, foliar area (limb and total) and foliar nitrogen concentration, although it not differing between 10 or 15g of N plant-1. For the accumulation of the nutrients Ca, Mg, S, Fe and Zn, the nitrogen fertilization did not represent higher foliar levels, even though this might be related to a dilution effect. / O mirtileiro é uma cultura recentemente introduzida no Brasil, sendo que várias pesquisas tem-se voltado para seu melhor desenvolvimento e adaptação no país. Por possuir poucos pelos radiculares, forma associações com microrganismos afim de aumentar sua zona de absorção de nutrientes, sendo o nitrogênio o elemento absorvido em maior quantidade. Neste trabalho, observou-se a interação dessa cultura com os inóculos Azospirillum brasilense e Trichoderma asperellum em diferentes substratos ou doses de adubação nitrogenada. O experimento foi realizado com a cv. Clímax e as plantas acondicionadas em vasos, em casa de vegetação, sob delineamento experimental de blocos casualizados em esquema fatorial 3x3. Os fatores analisados foram inoculação (A. brasilense, T. asperellum ou sem inoculação) e formulação dos substratos (5; 20 e 35% de matéria orgânica – esterco bovino curtido) ou, no segundo experimento, inoculação (A. brasilense, T. asperellum ou sem inoculação) e doses de nitrogênio (0, 10 e 15 g de N planta-1 – fornecido como ureia). A maior proporção de matéria orgânica propiciou as maiores médias em relação a variáveis fisiológicas (transpiração, eficiência no uso da água e eficiência de carboxilação) e de crescimento (área de limbo foliar, área foliar total e taxa de crescimento relativo), embora, para outras, não tenha diferido das plantas cultivadas com 20% de matéria orgânica. Com 35% de M.O., a inoculação com T. asperellum apresentou médias superiores em assimilação de CO2 (A), eficiência de carboxilação (A/Ci) e taxa de crescimento relativo (TCR). As doses de nitrogênio influenciaram positivamente A/Ci, teores de clorofila, área foliar (de limbo e total) e concentração foliar de nitrogênio, embora não diferindo entre 10 ou 15g de N planta-1. Para o acúmulo dos nutrientes Ca, Mg, S, Fe e Zn, a adubação nitrogenada não representou maiores teores foliares, embora isto possa estar relacionado a um efeito de diluição.

<p>U ovom radu su ispitani hemijski sastav, antioksidativna, antihiperglikemijska, antiproliferativna i antimikrobna aktivnost liofiliziranih metanolnih ekstrakata tropa<br />maline (Rubusidaeus L) sorti Meeker (ETMM) i Willamette (ETMW), kupine (RubusfruticosusL) sorti Čačanska bestrna (ETKČ) i Thornfree (ETKT) i divlje borovnice<br />(Vacciniummyrtillus L) (ETB). Sadržaj ukupnih polifenola (TPh), flavonoida (TF) i antocijana (TAc) u ekstraktima tropova određeni su spektrofotometrijskim metodama.<br />Identifikacija i kvantifikacija fenolnih jedinjenja (fenolnihkiselina, flavonola i cijanidina) i vitamina C u ekstraktima tropova izvedena je HPLC analizom. Spektrofotometrijskim<br />testovima određena je antioksidativna aktivnostna DPPH i ABTS+ radikale i redukciona sposobnost ekstrakata. ESR spektroskopijom utvrđen je uticaj ekstrakata na reaktivne hidroksil i superoksid anjon radikale. Antihiperglikemijska aktivnost ekstrakata određena<br />je testom inhibicije &alpha;-glukozidaze. Antiproliferativna aktivnost ekstrakata tropova ispitana je in vitro, njihovim delovanjem na rast četiri histolo&scaron;ki različite humane ćelije:<br />HeLa (epitelni karcinom cerviksa), MCF7 (adenokarcinom dojke), HT-29 (adenokarcinom debelog creva) i MRC-5 (humani fetalni fibroblasti pluća). Antimikrobna aktivnost ekstrakata tropova bobičastog voća ispitana je na odabrane sojeve Gram-pozitivnih i Gram-negativnih bakterija, kvasaca i plesni disk difuzionom metodom i metodom &bdquo;bunarčića&rdquo;. Ispitana je i mogućnost primene liofiliziranih ekstrakata tropova bobičastog voća kao funkcionalnog dodatka sokovima od voća. Laboratorijski proizvedenim obogaćenim sokovima je određen sadržaj TPh, TF i TAc i utvrđena antioksidativna aktivnost testom redukcione sposobnosti i DPPH testom. Rezultati ispitivanja hemijskog sastava, antioksidativne, antihiperglikemijske, antiproliferativne i antimikrobne aktivnosti ETMM, ETMW, ETKČ, ETKT i ETB ukazuju da su svi ispitivani ekstrakti tropova potencijalni izvor prirodnih<br />bioaktivnih jedinjenja, a antioksidativna aktivnost obogaćenih sokova ukazuje na mogućnost kori&scaron;ćenja ovih ekstrakata, kao funkcionalnih aditiva u proizvodnji različitih<br />visoko vrednih prehrambenih proizvoda.</p> / <p>This work examined the chemical composition, the antioxidant,<br />antihyperglycaemic, antiproliferative and antimicrobial<br />activity of the freeze-dried raspberry (Rubus idaeus L.)<br />varieties Meeker (ETMM) and Willamette (ETMW), blackberry<br />(Rubus fruticosus L.) varieties (ETKČ) and Thornfree<br />(ETKT), and wild blueberry (Vaccinium myrtillus L.) (ETB)<br />pomaces methanol extracts. The content of total polyphenols<br />(TPh), total flavonoids (TF) and total anthocyanins (TAc) in<br />the pomace extracts were determined spectrophotometrically.<br />Identification and quantification of the phenolic compounds<br />(phenol acids, flavonols and cyanidins) and ascorbic acid in<br />the pomace extracts was performed by HPLC analysis. The<br />antioxidant activities of the extracts was determined spectrophotometrically<br />using DPPH and ABTS+ free radicals and<br />reducing ability assay. ESR spectroscopy is used for determination<br />of pomace extracts&rsquo; influence on reactive hydroxyl<br />and superoxide anion radicals. Antihyperglycaemic activity of<br />the pomace extracts was determined using &alpha;-glucosidase inhibition<br />assay. The antiproliferative activity of the pomace extracts<br />was tested in vitro, testing their influence on the growth<br />of four histologically different human tumor cells: HeLa<br />(cervix epithelioid carcinoma), MCF7 (breast adenocarcinoma),<br />HT-29 (colon adenocarcinoma) and MRC-5(human fetal<br />lung fibroblasts). Antimicrobial activity of berry pomace extracts was tested on selected strains of Gram-positive and<br />Gram-negative bacteria, yeasts and molds with the disc diffusion<br />method and the method of &ldquo;wells&rdquo;. The possibility of<br />freeze dried berry pomace extracts application as a fruit functional<br />additive was investigated as well. Laboratory produced<br />enriched juice was tested in terms of TPH, TF and TAc content,<br />reducing ability and DPPH scavenging ability as well.<br />The results of testing the chemical composition, the antioxidative,<br />antihyperglycaemic, antiproliferative and antimicrobial<br />activities of ETMM, ETMW, ETKČ, ETKT and ETB, indicate<br />that all investigated pomace extracts are a potential source of<br />natural bioactive compounds, and that the antioxidant activity<br />enriched juice indicates the possibility of using these extracts,<br />as a functional additive in the production of various high-value<br />food products.</p>

This thesis covers more than thirty years of the author's research into the Paleoamerican period of the Middle Atlantic Region of North America, including the last 19+ years of focused work on the Cactus Hill site (44SX202) and replication of the Paleoamerican occupation discovered there. Using a landform and geology based predictive model derived from the Paleoamerican occupation at Cactus Hill, the author directed preliminary archaeological testing in three other areas of the same Nottoway River Valley, where Cactus Hill is located. These areas were the Barr site, located 11 miles (18 km.) downriver from Cactus Hill; the Chub Sandhill Natural Resource Conservation Area, located 19 miles (30 km.) downriver from Cactus Hill; and the Blueberry Hill site (44SX327), located approximately 1,000 feet (300 meters) east of Cactus Hill. The latter two produced OSL dated, pre-Younger-Dryas landforms, as predicted. The Rubis-Pearsall site (44SX360), located in the Chub Sandhill preserve also produced a buried Paleoamerican, Clovis age cultural level confirming the model. In addition to the OSL dates, Blueberry Hill also produced a distinct and apparently discrete activity surface with a possible pre-Clovis age Cactus Hill point at the same depth as the Paleoamerican levels at Cactus Hill and Rubis-Pearsall.

Le diabète de type 2 est une maladie chronique dont l’incidence est en augmentation continuelle. Le risque de développer le diabète de type 2 chez les populations autochtones du Canada est de trois à cinq fois plus élevé que le reste de la population canadienne. La forêt boréale comporte plusieurs plantes médicinales ayant un potentiel pour le traitement ou la prévention du diabète. Certaines de ces plantes font partie de la médecine traditionnelle et alternative Crie. Des enquêtes ethnobotaniques ont amené notre équipe de recherche à identifier 17 extraits de plantes médicinales utilisées par les Cris d’Eeyou Istchee (Baie James, Québec) pour traiter les symptômes du diabète. Parmi ces extraits, certains ont montré des activités anti-diabétiques au niveau des cellules musculaires, des adipocytes et dans des études in vivo réalisées chez des animaux. Le but de cette thèse est d’élucider l’effet de ces 17 plantes sur l’homéostasie hépatique de glucose, d’identifier l’espèce la plus prometteuse et isoler ces constituants actifs. De même, le bleuet nain du genre Vaccinium angustifolium fait partie de la forêt boréale canadienne et est connu pour ses activités anti-diabétiques. Une biotransformation du jus de bleuet lui confère une activité antioxydante accrue et un profil biologique différent. Le deuxième but de cette thèse est d’élucider les mécanismes d’action par lesquels le jus de bleuet biotransformé (BJ) exerce son effet anti-diabétique et d’identifier ses principes actifs. Les résultats ont montré que trois extraits de plantes Cris se sont démarqués par leur effet sur l’homéostasie hépatique de glucose. Picea glauca exerce son effet en diminuant la production de glucose alors que Larix laricina agit en augmentant le stockage de glucose. Abies balsamea a montré le profil le plus prometteur, elle agit simultanément en diminuant l’activité de la Glucose-6-phosphatase (G6Pase) via la stimulation des voies insulino-dépendante et - indépendante et en augmentant l’activité de la Glycogène synthétase (GS) suite à la phosphorylation de la Glycogène synthase kinase-3. Le fractionnement de l’extrait d’Abies balsamea guidé par les deux bioessais a mené à l’isolation de trois composés actifs; l’acide abiétique (AA), l’acide déhydroabiétique (DAA) et le squalène (SQ). Les principes actifs ont montré le même mécanisme d’action que l’extrait brut en diminuant l’activité de la G6Pase et augmentant celle de la GS ainsi qu’en activant les voies de signalisation impliquées. Le DAA ii s’est démarqué par son effet le plus puissant et très comparable à celui de l’extrait d’Abies balsamea dans toutes les expériences. De son côté le BJ a montré un effet sur la diminution de la production hépatique de glucose, l’augmentation de son stockage ainsi que l’augmentation de son transport dans le muscle. Son fractionnement guidé par les bioessais a permis d’isoler sept fractions dont trois étaient les plus actives. L’identification des constituants de ces fractions actives a mené à isoler quatres composés phénoliques; l’acide chlorogénique, l’acide gallique, l’acide protocatéchique et le catéchol. Le catéchol s’est démarqué avec ses effets les plus puissants en diminuant l’activité de la G6Pase, augmentant celle de la GS et en stimulant le transport de glucose dans le muscle. Les résultats de cette thèse indiquent que la diminution de la production hépatique de glucose peut s’ajouter au profil anti-diabétique de certaines plantes médicinales Cries et surtout à celui d’A.balsamea dont les composés actifs peuvent aider dans le développement de nouvelles molécules anti-diabétiques. De plus, les résultats de cette thèse ont montré que l’activité antidiabétique du BJ implique le contrôle de l’homéostasie de glucose au niveau du foie et du muscle. L’identification du catéchol comme principe actif avec potentiel anti-diabétique prometteur pourra servir pour des fins thérapeutiques ultérieures. / Type 2 diabetes is a chronic disease for which incidence is continuously increasing. The risk of developing type 2 diabetes among Aboriginal people in Canada is three to five times higher than the rest of the Canadian population. The boreal forest has several medicinal plants with potential for the treatment or prevention of diabetes. Some of these plants are part of the Cree traditional and alternative medicine. Ethnobotanical surveys led our research team to identify 17 medicinal plant extracts used by the Crees of Eeyou Istchee (James Bay, Quebec) to treat symptoms of diabetes. Some extracts showed anti-diabetic activities in muscle cells, adipocytes and in vivo studies in animals. The aim of this thesis is to elucidate the effect of these 17 plants on hepatic glucose homeostasis, to identify the most promising species and isolate its active constituents. Similarly, Canadian lowbush blueberry, Vaccinium angustifolium.Ait, is part of the Canadian boreal forest and is known for its anti-diabetic activities. Biotransformation of blueberry juice gives it an increased antioxidant activity and a different biological profile. The second aim of this thesis is to elucidate the mechanisms of action by which biotransformed blueberry juice (BJ) exerts its anti-diabetic effect and identify its active constituents. The results showed that three Cree plants stood out with their effect on hepatic glucose homeostasis. Picea glauca exerts its effect by reducing glucose production whereas Larix laricina works by increasing its the storage. Abies balsamea showed the most promising profile, simultaneously and powerfully reducing glucose-6-phosphatase (G6Pase) involving both insulin-dependent and -independent pathways and stimulating Glycogen synthase (GS) via phosphorylation of Glycogen synthase kinase-3 (GSK-3). Bioassay-guided fractionation of Abies balsamea led to the isolation of three active compounds; Abietic acid (AA), dehydroabietic acid (DAA) and squalene (SQ). The active constituents have shown the same mechanism of action as the crude extract by decreasing the activity of G6Pase, increasing that of the GS and activating signaling pathways. DAA stood out for its most powerful effect close to that of the crude extract in all experiments. Our results showed that anti-diabetic activity of BJ involves decrease in hepatic glucose production, increase of storage and enhancement of glucose uptake in muscle. Its bioassayiv guided fractionation led to isolate seven fractions, three of which were the most active. Identification of components in the active fractions resulted in four isolated phenolic compounds; chlorogenic acid, gallic acid, protocatechuic acid and catechol. Catechol stood out with its most powerful effects by decreasing the activity of G6Pase, increasing the GS and stimulating glucose transport in muscle. Our results thus confirm that the reduction of hepatic glucose production likely contributes to the therapeutic potential of several anti-diabetic Cree traditional plant and especially that of Abies balsamea whose active compounds may help in the development of new anti-diabetic molecules. In addition, the results of this thesis showed that the anti-diabetic activity of BJ involves control of glucose homeostasis in the liver and muscle. Identification of catechol as an active compound with anti-diabetic promising potential can be used for future therapeutic purposes.

Le bleuet sauvage (Vaccinium angustifolium Ait. et V. myrtilloides Michaux) représente un marché en plein essor au Canada, premier pays producteur et exportateur mondial de ce fruit. Pour faire face à la demande, les producteurs cherchent continuellement à adapter leurs pratiques de production dans le but d’améliorer leur rendement et l'état de santé de leurs bleuetiers. Or, les micro-organismes présents dans les racines et dans le sol jouent un rôle non négligeable en lien avec la santé des plantes. Ce microbiote est donc d’intérêt d’un point de vue agronomique, pourtant, contrairement à d’autres cultures, très peu d’études se sont penchées spécifiquement sur le microbiote du milieu racinaire du bleuetier sauvage. Ce doctorat s’inscrit donc dans l’optique d’accroître les connaissances sur les communautés bactériennes et fongiques présentes dans les bleuetières au Québec. Les objectifs de ce projet sont de détecter les taxons qui pourraient avoir un impact sur les variables agronomiques des bleuetiers telles que le rendement; d’identifier les variables physico-chimiques du sol influençant ces communautés; et d’étudier les impacts que peuvent avoir les différentes pratiques agricoles, telles que la fertilisation et la fauche thermique, sur ces micro-organismes. Nous nous sommes appuyés sur le séquençage de nouvelle génération et le métacodage à barres de l’ADN environnemental de nos échantillons de racines et de sol afin d’obtenir une analyse des communautés bactériennes et fongiques de la rhizosphère et des racines des bleuetiers. Les analyses multivariées effectuées par la suite permettent de comparer ces communautés et de voir si certaines espèces sont spécifiques à une condition particulière. Dans l’ensemble, cette thèse a donc permis de caractériser les communautés fongiques et bactériennes du milieu racinaire du bleuetier sauvage in situ dans plusieurs bleuetières du Québec. De nombreuses espèces de champignons mycorhiziens éricoïdes ont été systématiquement identifiées dans les trois études et leur prédominance suggère leur importance pour le bleuetier sauvage. Nous avons également trouvé que l’ordre bactérien des Rhizobiales, connu pour sa capacité à fixer l’azote atmosphérique, occupait une part importante de la communauté bactérienne. Les études sur la fertilisation et la fauche thermique ont démontré que ces deux pratiques agricoles avaient peu d’impact significatif sur les communautés microbiennes étudiées. Enfin, cette thèse donne des pistes de réflexion sur la fixation d’azote par les communautés bactériennes et pose les premières bases pour des essais de bio-inoculation avec les espèces fongiques et bactériennes détectées ayant un potentiel impact bénéfique sur la culture des bleuets sauvages. / The wild blueberry (Vaccinium angustifolium Ait. and V. myrtilloides Michaux) market is booming in Canada, the world's leading producer and exporter of this fruit. In order to meet the demand, growers are constantly trying to adapt their production practices to improve their yields and the health of their blueberry fields. Micro-organisms present in the roots and in the soil play a significant role in the health of the plants. This microbiota is therefore of interest from an agronomic point of view, yet, contrary to other crops, very few studies have been conducted specifically on the microbiota of the root environment of wild blueberries. This doctoral project therefore aims at increasing our knowledge of the bacterial and fungal communities present in wild blueberry fields in Quebec. The objectives of this project are to detect taxa that could have an impact on agronomic variables of wild blueberry fields such as fruit yield; to identify soil physico-chemical variables influencing these communities; and to study the impacts that different agricultural practices, such as fertilization or thermal pruning, may have on these micro-organisms. We relied on next generation sequencing and metabarcoding of environmental DNA from our root and soil samples to obtain an analysis of the bacterial and fungal communities in the rhizosphere and roots of blueberry shrubs. Subsequent multivariate analyses allow us to compare these communities and see if certain species are specific to a particular condition. Overall, this thesis has characterized the fungal and bacterial communities in the root environment of wild blueberry in situ in several Quebec wild blueberry fields. Numerous species of ericoid mycorrhizal fungi were systematically identified in all three studies, and their predominance suggests their importance to wild blueberries. We also found that the bacterial order Rhizobiales, known for its ability to fix atmospheric nitrogen, occupied an important part of the bacterial community. Studies on fertilization and thermal mowing showed that these two agricultural practices have limited significant impacts on the microbial communities studied. Finally, this thesis provides insights into nitrogen fixation by bacterial communities and lays the groundwork for bio-inoculation trials with the fungal and bacterial species detected to have a potential beneficial impact on wild blueberry cultivation.