Colletotrichum gloeosporioides is the causal agent of anthracnose disease of mangoes. Infection occurs when humidity is high and rain-dispersed spores germinate and form an appressorium on immature mangoes. The infection then becomes quiescent until the fruit is harvested. On ripe fruit infection is visible as black, sunken lesions on the surface. At the pre-harvest stage, the disease is controlled with the application of a range of fungicides, and at the post-harvest stage by hot benomyl treatment. The extensive use of benomyl, both pre- and post-harvest, has resulted in the occurrence of isolates of C. gloeosporioides resistant to this fungicide. To devise an alternative strategy of disease control, the potential for biological control of anthracnose has been investigated. Potential microbial antagonists of C. gloeosporioides were isolated from blossom, leaves and fruit of mango, and screened using a series of assay techniques. In total 650 microorganisms, including bacteria, yeasts and filamentous fungi, were isolated and tested for their inhibition of growth of C. gloeosporioides on malt extract agar. Of these 650 isolates, 121 inhibited the fungus and were further tested on their ability to inhibit spore germination in vitro. Of these, 45 isolates, all bacteria and yeasts, were inoculated onto mangoes, which were artificially inoculated with C. gloeosporioides, and assessed for their potential to reduce the development of anthracnose lesions. A further selection was made, and 7 isolates were chosen to be used in a semi-commercial trial in the Philippines. This final screening procedure yielded two potential candidates for field trials, isolate 204 (identified as Bacillus cereus) and isolate 558 (identified as Pseudomonas fiuorescens). A field trial involving pre-harvest application of the biological control agent, was conducted using isolate 558. This isolate was chosen for this purpose since in in vitro experiments it significantly reduced germination of C. gloeosporioides spores. In the field trial 558 was applied in combination with nutrients and compared to treatments which had received no treatment or which had received conventional fungicide (benomyl) application. On spraying, high numbers of 558 were recorded on the leaf surface, but no reduction in post-harvest development of disease was observed. Failure of disease control was attributed to rapid death of the bacterium on the phylloplane. Inpost-harvest trials, isolates 204 and 558 were both tested in combination with different application methods, including the addition of sticker, peptone, fruit wax or a sucrose polyester. Application of 204 did not reduce disease development. Application of 558, however, did significantly reduce anthracnose development compared to the control fruit. No additional benefit was achieved by incorporating the bacteria in peptone, fruit wax or sucrose polyester. The mode of action of isolate 558 was investigated in detail. There was no evidence for parasitism taking place, or the production of volatile compounds, in the suppression of disease development. No antibiotic compounds were detected, but isolate 558 did produce a siderophore. A sharp increase in pH was also observed in culture media in which 558 was grown. Disease control may result from a combination of these two factors.

Two endemic late-season grape rot diseases, ripe rot and sour rot continue to pose a significant challenge to grape growers in Virginia and similar climates. Managing these diseases in the vineyard is challenging due to concerns over the increased risk of resistance against traditional single-site mode of action pesticides. This study evaluated the efficacy of different biopesticides and novel chemical options with a series of field trials across four vineyards in northern Virginia in 2021 and 2022. Among the tested products, a potassium-based nutrient formulation resulted in consistent ripe rot suppression in two of the three trials, showing around 46% and 83% less mean disease severity at the AHS AREC trial in 2022 and 2021, respectively, and 63% and 19% less mean disease incidence at the South Loudoun trial in 2021 and AHS AREC trial in 2022. For sour rot, zeta-cypermethrin with cyprodinil + fludioxonil or hydrogen peroxide + peroxyacetic acid resulted in effective control in two of the five trials, with a high of 71% and 73% less mean disease incidence compared to the untreated check in the Frederick trial in 2021 and 2022, respectively. In general, treatments with spinosad were less effective and resulted in lower disease control compared to those with zeta-cypermethrin and its fungicides/antimicrobials counterpart. The results suggest that integrating these biopesticides and novel chemicals into a spray program could offer a more sustainable solution for sour rot and ripe rot disease management without compromising the level of disease control. / Master of Science in Life Sciences / Ripe rot and sour rot are two diseases of concern for growers in Virginia and many other grape-growing regions due to significant losses in fresh cluster yield and the risk of contamination during the winemaking process. Traditional pesticides are often used; however, the breakdown of the effectiveness of these pesticides after continuous application, i.e., pesticide resistance, has become a critical issue. The other issues include, but are not limited to, adverse effects on human health, nature, and winemaking. On the other hand, biopesticides derived from microorganisms, their products, plant extracts, etc., are considered relatively safer and more sustainable options. This study evaluated the efficacy of biopesticides and novel chemicals against ripe rot and sour rot through field trials over two years at four locations in northern Virginia. Although no clear-cut winners were among the tested treatments, one of the tested chemicals (a potassium-based plant nutrient formulation) provided consistent disease suppression (up to 83%) against ripe rot. In the case of sour rot, several treatments, including a biologically derived insecticide, produced comparable disease suppression (up to 73%) compared to the current standard application. These safer options can be included in the spray schedule without compromising the level of disease control, meanwhile aiding the sustainability of the spray program in the long run.

Ripe rot of grape is caused by two fungal species complexes: <i>Colletotrichum gloeosporioides</i> and <i>C. acutatum</i>, both of which cause disease on a variety of crops, such as strawberry and apple. To investigate effect of cultivar and cluster developmental stage on the development of ripe rot, controlled environment and field studies were conducted during 2013-2014. We have identified that a certain level of infection could can take place on most cultivars tested from bloom to the near harvest. In most of the cases, significant cultivar and cluster developmental stage interaction effects were observed (<i>P</i> < 0.05) for the development of disease symptoms in both studies. In general, susceptible cultivar (Cabernet Franc, Cabernet sauvignon, and Chardonnay) demonstrated fluctuations of disease susceptibility among cluster development stages, while resistant cultivars (Merlot) showed consistently low level of the disease throughout the season. To investigate the effect of eleven ten modes of action for control of <i>C. gloeosporioides</i> and <i>C. acutatum</i>, two methods, alamarBlue® assay and inoculation on fungicide-treated detached fruits, were used. Protective fungicides (mancozeb, captan, and copper) as well as some of newer formulations such as azoxystrobin and tetraconazole were identified as excellent products against ripe rot of grape. Four additional materials were identified as good potential candidates to investigate further. The information gained from these studies will help growers to determine the critical period for ripe rot management and chemicals to be applied for management. With better control of cluster rot pathogens, Virginia growers can experience an increase in yield and wine quality. / Master of Science in Life Sciences

IPv4-adresser är begränsat när det gäller antalet det finns att dela ut, ett framtida problem är att dessa adresser med största sannolikhet kommer att ta slut. Vårt arbete har således fokuserat på att inhämta information som bidrar till en klarare bild av hur dagsläget ser ut kopplat mot implementeringen av IPv6 som sker i Europa och Sverige. För att kunna förklara denna implementering på ett bra sätt finns det delar i arbetet som beskriver de tekniska hinder och möjligheter som bidrar till införandet av detta protokoll. Huvuddelen av tiden har spenderats med att inhämta information och fakta som ger en fördjupad kunskap i ämnet, med avsikten att skriva en gedigen bas som utredningsdelen i arbetet utger. Intervjuer har också varit en del, denna del finns i syfte att ge en bild över hur internetleverantörer arbetar med införandet av IPv6. De organisationer som finns och arbetar med införandet av IPv6 har alla en bra pool med fakta som beskriver hur situationen ser ut i dagsläget. RIPE NCCs databas visar klart och tydligt de organisationer som finns med i arbetet med implementeringen av IPv6. Denna databas samt information tagen från genomförda intervjuer ger oss en uppfattning om att införandet än så länge inte har kommit särskilt långt, detta i relation till bristen av IPv4 adresser som råder. I Sverige jobbar PTS mycket mot att hjälpa organisationer med en eventuell övergång till IPv6, deras arbete är ordentligt men problematiken där är istället att organisationer idag inte är i behov av denna information. Det positiva är således att informationen finns där, hos PTS, att hämta vid behov.Trots att de sista IPv4-blocken är utdelade finns det egentligen inget behov av att implementera IPv6 i dagsläget. Ett IPv4-nätverk med fungerande NAT fungerar och kommer med största sannolikhet fungera ett bra tag framöver.

A oxidação lipídica é uma das causas de deterioração e perda de qualidade dos alimentos. Com a intenção de reduzir ou inibir essa reação, são empregados antioxidantes sintéticos. No entanto, com a crescente preocupação sobre o efeito dos mesmos sobre a saúde, diversas pesquisas têm por objetivo buscar alternativas de fontes naturais. A atividade antioxidante de frutas tem sido muito estudada. A acerola se destaca devido aos seus componentes, como o ácido ascórbico, compostos fenólicos, carotenoides e antocianinas, conferindo elevada capacidade antioxidante, principalmente no estádio de maturação verde, o qual pode ser explorado como base para aditivo para a indústria alimentícia. A técnica de microencapsulação pode facilitar a aplicação, proteger os compostos ativos e favorecer sua liberação controlada em um produto. O objetivo deste trabalho foi obter extratos de frutos de acerola em condições otimizadas e submetê-los ao processo de microencapsulação, visando avaliar seu potencial de aplicação como antioxidante em emulsão lipídica. Foi realizada a otimização da extração de compostos ativos de amostras de frutos verdes e maduros de acerola por meio da metodologia de Superfície de Resposta, em que foram determinados os efeitos da proporção amostra:solvente e tempo sobre a atividade antioxidante (DPPH e ABTS) e compostos fenólicos totais (CFT). Os extratos obtidos em condições otimizadas foram caracterizados quanto ao teor de compostos fenólicos totais (CFT), atividade antioxidante (DPPH, ABTS, ORAC-H e FRAP), teores de ácido ascórbico e compostos fenólicos (clorogênico, ferúlico e p-cumárico). O extrato com maior atividade antioxidante foi selecionado para a obtenção de micropartículas por spray drying, com adição de goma arábica e maltodextrina. As micropartículas foram caracterizadas (umidade, aw, solubilidade, higroscopicidade, tamanho de partículas), avaliadas por microscopia eletrônica de varredura (MEV) e também quanto à manutenção da atividade antioxidante. As micropartículas, o extrato aquoso de acerola otimizado e o antioxidante sintético TBHQ (Terci-Butil-Hidroquinona) foram aplicados em sistema modelo de emulsão em concentrações de até 200 mg. Kg-1. As emulsões foram submetidas ao teste acelerado de oxidação a 40°C por 9 dias, com acompanhamento da variação do teor de hidroperóxidos, dienos conjugados e índice de cremeação (CI) a cada 3 dias. A otimização propiciou a obtenção de extratos com elevada atividade antioxidante. Para os frutos maduros a melhor proporção amostra:solvente (água) foi de 1:30, já para frutos verdes foi de 1:40. O tempo ótimo identificado foi 10 minutos de extração. Os extratos de frutos verdes apresentaram atividade antioxidante superior aos de frutos maduros, bem como elevadas concentrações de ácido ascórbico e ácidos fenólicos. As micropartículas apresentaram baixa umidade e atividade de água. A maior solubilidade e menor higroscopicidade foi verificada nas produzidas com maltodextrina. Ambas apresentaram partículas pequenas e características tecnológicas adequadas para aplicação em alimentos; entretanto a micropartícula produzida com goma arábica se destacou pelo elevado rendimento no processo e proteção da atividade antioxidante. Os tratamentos com a adição de micropartículas em emulsões apresentaram efeito protetor significativo e semelhante ao antioxidante sintético TBHQ nas concentrações de 100 e 200 mg.kg-1 na avaliação da oxidação lipídica. No entanto, o efeito protetor conferido pela adição de micropartículas com goma arábica e de mistura de micropartículas a 200 mg.kg-1 permitiram maior controle do processo oxidativo, com a observação de baixos teores de hidroperóxidos, dienos conjugados e menores índices de cremeação durante o teste acelerado. Conclui-se que extratos microencapsulados de frutos verdes de acerola são promissores para aplicação em emulsões lipídicas e apresentam potencial para substituir antioxidantes sintéticos em alimentos e cosméticos. / Lipid oxidation is one of the major causes of deterioration and loss of food quality. Synthetic antioxidants are applied in order to reduce or inhibit this reaction. However, with growing concern about health aspects of these antioxidants, several research aims to find alternatives from natural sources. The antioxidant activity in fruits has been studied. Acerola stands out due to its components, such as ascorbic acid, phenolic compounds, carotenoids and anthocyanins, that may are related to high antioxidant capacity, specially at the green maturation stage, which can be exploited as an additive for the food industry. The microencapsulation technique may facilitate its apllication by protecting the compounds and promoting controlled release of active compounds. The objective of this work was to obtain extracts of acerola fruit under optimized conditions and to submit them to the microencapsulation process, aiming to evaluate its potential application as antioxidant in lipid emulsion. The optimization of the extraction of active compounds from green and ripe acerola fruits was achieved through response surface methodology, in which the effects of proportion sample:solvent and time on the antioxidant activity (DPPH and ABTS) and total phenolic compouns (TPC) were determided. The extracts obtained under optimized conditions were characterized as the content of total phenolic compounds (TPC), antioxidant activity (DPPH, ABTS, ORAC-H and FRAP), ascorbic acid and phenolic compounds (chlorogenic, ferulic, and p-coumaric acids). The extract with greater antioxidant activity was selected to obtain microparticles by spray drying, with addition of gum arabic and maltodextrin. The microparticles were characterized (moisture, water activity, hygroscopicity, solubility, particle size), evaluated by scanning electron microscopy (SEM) and the maintenance of the antioxidant activity. The microparticles, the optimized aqueous acerola extract and the synthetic antioxidant TBHQ (terc-butyl-hydroquinone) were applied in model system of emulsion in concentrations up to 200 mg. Kg-1. The emulsions were subjected to accelerated oxidation test at 40°C for 9 days, with monitoring of the variation of the hydroperoxides content, conjugated dienes and creaming index (CI) every 3 days. The optimization promoted to obtention of extracts with high antioxidant activity. For the ripe fruits the best sample:solvent (water) ratio was 1:30 and for green fruits it was 1:40. Optimum time identified was 10 minutes of extraction. The extracts of green fruits showed higher antioxidant activity than mature fruits, as well as higher concentrations of ascorbic acid and phenolic acids. The microparticles had low moisture content and water activity. The greater solubility and lower hygroscopicity was observed in those produced with maltodextrin. Both showed small particles and technological characteristics suitable for application in food; however the microparticle produced with gum arabic was distinguished by the high yield in the process and protection of the antioxidant activity. The treatments with addition of microparticles in emulsions showed significant protective effect similar to the synthetic antioxidant (TBHQ) at concentrations of 100 and 200 mg.kg-1 in the evaluation of lipid oxidation. However, the protective effect conferred by the addition of microparticles with gum arabic and a mixture of microparticles at 200 mg.kg-1 allowed the greater control of the oxidative process, with the observation of low hydroperoxides content, conjugated dienes and lower creaming inde during the accelerated test. It is concluded that microencapsulated extracts of green acerola fruits are a promising for application in lipid emulsions and show a potential to replace synthetic antioxidants in food and cosmetics.

Tato práce se zabývá problematikou zjišťování zpětných cest v Internetu. Nástroj, který by byl schopen určit zpětnou cestu, by mohl být cenný v například v případech, kdy určitá část zákazníků pozoruje zvýšenou latenci při využívání služby. Klasickým nástrojem pro analýzu cesty k cílovému počítači je traceroute. Práce se detailně zabývá diagnostickým nástrojem traceroute a jsou diskutovány nejen jeho rozšíření, ale také nedostatky v sítích, kde se vyskytuje vyvažování provozu, a jejich možná řešení. Nicméně, pokud se problém nachází ve směru od zákazníků k poskytovateli služby, pak odhalení problému může být problematické. Dále je studován existující výzkum v oblasti zjišťování zpětných tras v Internetu a nástroje pro diagnostiku sítě. Součástí práce je navržení a implementace nástroje, který je schopen aproximovat zpětnou cestu s využitím vhodné RIPE Atlas sondy a získaná data dále analyzovat. Implementovaný nástroj byl testován na vytvořené topologii i v reálném provozu s využitím referenčního virtuálního serveru.

This thesis is focused on finding the physical location of stations by the passive geolocation techniques, particularly using the registration database of IP addresses. The first two part are focused on a theoretical analysis of this problem, the next two parts of this thesis deal with the solution of this problem and discussion of the results.

Diploma thesis deals with monitoring and diagnostics of transport networks. It focuses on basic diagnostic and surveillance tools, and tools which are developed in the project of Internet2. It is focused on the evaluation of the measurements performed by these tools with a focus on external factors and substandard conditions and their impact on the measurement results.

Colletotrichum acutatum and C. gloeosporioides are fungal plant pathogens that have a global distribution, extensive host range, and convoluted taxonomy. Both species can cause grape ripe rot and are considered endemic to Virginia US. In 2012, C. acutatum and C. gloeosporioides were reclassified into species complexes that consist of 31 and 22 accepted species, respectively. The objectives of this study were to: 1) survey Virginia vineyards for grape ripe rot, and morphologically and phylogenetically identify isolates to the species within the complexes, 2) conduct an in vitro fungicide assay to screen fifteen commercial fungicides and combinations of two fungicides for efficacy to control isolates from seven Colletotrichum species from Virginia vineyards, 3) sequence gene fragments from three subunits of the SDH enzyme in the fungicide-screened isolates to observe potential resistance mutations, 4) investigate the susceptibility of three grapevine tissues to Colletotrichum species, 5) observe potential infection structures before and after the application of fungicides, 6) evaluate the efficacy of commercial fungicide controls of grape ripe rot in the field, and determine the most advantageous timing of applications. In my studies, I identified six Colletotrichum species: C. aenigma, C. conoides, C. fioriniae, C. gloeosporioides, C. kahawae, and C. nymphaeae. I also found two additional groups; an isolate similar to C. limetticola and C. melonis and a group of isolates that are similar to C. alienum, C. fructicola, and C. nupharicola. I also identified captan, and mancozeb as two potential active ingredients for control of grape ripe rot isolates from Virginia via the in vitro fungicide assay. Additionally, I found that combinations of two active ingredients could increase the efficacy of benzovindiflupyr, copper, and polyoxin-D. C. fioriniae germination and production of melanized appressoria was documented on leaves. I observed appressorium formation with isolates of two C. fructicola-like genotypes and C. nymphaeae, as well as secondary conidiation with isolates of C. aenigma, C. fructicola-like genotype 3, and C. nymphaeae on blooms. And finally, benzovindiflupyr, cyprodinil + fludioxonil pre-mix, and potassium phosphite + tebuconazole were identified as candidates for chemical control for grape ripe rot in the field. / PHD / Colletotrichum acutatum and C. gloeosporioides are two fungal plant pathogens that are found on a wide range of crops around the globe. Both fungal species cause the disease grape ripe rot and have been found in Virginia (VA) USA since the late 1800s. Originally, grape ripe rot was considered a minor disease in VA; however, based on communications with local VA vineyard managers, grape ripe rot was found to cause up to 30% direct crop loss. Further indirect economic loss occurs during wine production due to the production of unpalatable, tobacco-like, off flavors from the infected grapes. Sensory studies found this wine flavor change occurred with as little as 3% of the total crushed grapes being infected. Grape ripe rot appears as a sunburn-like, tan injury on the surface of white-fruited grape berries. As the disease progresses, the dark injury expands across the surface of the berry and rings of salmon-colored spore masses form. On red-fruited grapes, the formation of spore masses is usually the first observable sign. Over time, the infected berries will shrivel down to a soft, pustule-covered raisin. Both C. acutatum and C. gloeosporioides cause the same grape ripe rot symptoms on fruit and overlap in fungal appearance. In addition, investigations of these pathogens using molecular techniques have revealed that each consists of a number of genetically distinct groups that are difficult to distinguish by appearance. Therefore, in 2012, C. acutatum and C. gloeosporioides were reclassified into 31 and 22 newly accepted species, respectively, using molecular techniques. The objectives of this study were to: 1) survey VA vineyards for grape ripe rot, and visually and molecularly identify isolates to the species within the new complexes, 2) conduct a laboratory fungicide assay to screen fifteen commercial fungicides and combinations of two fungicides for control of isolates from VA vineyards, 3) sequence gene fragments from three subunits of the SDH enzyme in the fungicide-screened isolates to observe potential resistance mutations, 4) investigate the susceptibility of three grapevine tissues to Colletotrichum species, 5) observe potential infection structures before and after the application of fungicides, 6) evaluate the efficacy of commercial fungicide controls of grape ripe rot in the field, and determine the most advantageous timing of applications. In my studies, I identified six Colletotrichum species: C. aenigma, C. conoides, C. fioriniae, C. gloeosporioides¸ C. kahawae, and C. nymphaeae. I also found two additional groups; an isolate similar to C. limetticola and C. melonis and a group of isolates that are similar to C. alienum, C. fructicola, and C. nupharicola. Our lab also identified four active ingredients as potential controls of grape ripe rot in the laboratory fungicide assay; captan, mancozeb, tetraconazole and thiophanate-methyl. Additionally, combinations of two compounds can increase the effectiveness of benzovindiflupyr, copper, and polyoxin-D. of C. fructicola-like isolates, and C. nymphaeae formed infection structures on blooms. of C. aenigma, C. fructicola-like genotype 3, and C. nymphaeae formed spores on blooms without producing symptoms. C. fioriniae spores germinated and produced infection structures on leaves without producing symptoms.

Countries that have experienced civil war suffer a greater risk for new conflict than countries with no prior history of civil war. This empirical finding has been called a conflict trap where the legacy of previous war - unsolved issues, indecisive outcomes, and destruction – leads to renewed fighting. Yet, countries like Cambodia, El Salvador, Indonesia, and Mozambique have managed to overcome decade-long conflicts without relapse. This dissertation addresses this empirical puzzle by seeking to dismantle the conflict trap and look at microlevel explanations for civil war resolution and relapse. It adds to existing scholarship in three ways: first, by using disaggregated empirics on war termination and how fighting resumes; second, by exploring government agency in conflict processes; and third, by disaggregating rebel organizations. Essay I present original data on the start and end dates and means of termination for all armed conflicts, 1946-2005. Contrary to previous work, this data reveal that wars does not always end through victory or peace agreement, but commonly end under unclear circumstances. Essay II addresses how developments exogenous to the conflict influence governments’ decision to engage in a peace process. The results show that after natural disasters when state resources need to be allocated towards disaster relief, governments are more willing to negotiate and conclude ceasefires with insurgents. Essay III focuses on the post-conflict society, and posits that security concerns among former war participants will push them towards remobilizing into rebellion. The findings indicate that if ex-belligerent elite’s security is compromised, the parties of the previous war will resume fighting, while insecurity among former rank-and-file leads to the formation of violent splinter rebel groups. Finally, Essay IV seeks to explain why governments sometimes launch offensives on former rebels in post-conflict countries. The results show that internal power struggles provide leaders with incentives to use force against domestic third parties to strengthen their position against intra-government rivals. Taken together, this dissertation demonstrates that there is analytical leverage to be had by disaggregating the processes of violence in civil war and post-conflict societies, as well as the actors involved – both the government and rebel sides.