Study of population dynamics of bacteria associated with pine wood nematode after inoculation with different strains of Bursaphelenchus xylophilus in maritime pine (Pinus pinaster)

Durante algum tempo pensou-se que o Bursaphelenchus xylophilus era o único agente etiológico da doença do nemátode da madeira do pinheiro. Recentemente, descobriu-se que existem bactérias associadas ao nemátode que contribuem para a patogénese desta doença, sobretudo através da libertação de toxinas que promovem a morte dos pinheiros. De entre as espécies mais comummente encontradas estão bactérias pertencentes aos géneros Bacillus, Pantoea, Pseudomonas e Xanthomonas. Este trabalho teve como principal objectivo o estudo do efeito da inoculação de pinheiro bravo (Pinus pinaster) com quatro isolados diferentes de nemátodes, na população bacteriana dos nemátodes e das árvores, em diferentes fases da progressão da doença. A monitorização da progressão dos sintomas da doença foi igualmente registada. Pretendeu-se também identificar as bactérias isoladas do xilema das árvores e da superfície dos nemátodes através de métodos de identificação clássicos, do sistema de identificação API20E e da sequenciação de ADN bacteriano. Os resultados obtidos demonstraram que, relativamente à progressão dos sintomas da doença, a diferença mais marcante foi verificada para os pinheiros inoculados com o isolado não virulento C14-5, onde se visualizou um agravamento de sintomas mais lento e menos severo do que nos pinheiros inoculados com os isolados virulentos. Verificou-se que numa fase inicial da doença, no geral, a população bacteriana dos ramos inoculados foi menor do que a que foi verificada passados 7 e passados 14 dias de inoculação. Num dos métodos de quantificação foi recuperado um maior número de bactérias nos pinheiros inoculados do que nos controlos aos 7º e ao 14º dias de inoculação. Os isolados que levaram à obtenção de maior quantidade bacteriana foram o HF e o 20, sendo que, comparativamente, o isolado não virulento levou à obtenção de menor quantidade bacteriana. Relativamente à identificação bacteriana, concluiu-se que o sistema API20E não foi suficiente na identificação das espécies bacterianas isoladas, mostrando-se pouco discriminatório, tendo sido identificada a espécie Enterobacter cloacae em 79% dos isolados e não sendo possível a identificação de sete colónias bacterianas. Assim, a adopção de métodos moleculares de identificação, através da sequenciação do ADN bacteriano, permitiu uma identificação mais fiável, tendo sido identificadas onze espécies bacterianas diferentes dentro dos géneros Klebsiella, Bacillus, Enterobacter, Paenibacillus, Terribacillus, Citrobacter, Pantoea e Escherichia. No geral, a diversidade bacteriana aumentou ao longo da progressão da doença. A espécie Bacillus spp. predominou na fase mais precoce da doença e a espécie Klebsiella oxytoca nas fases mais tardias. As espécies bacterianas isoladas da superfície dos nemátodes não diferiram muito das isoladas do xilema dos pinheiros. Neste trabalho foram identificadas espécies bacterianas nunca antes reportadas neste tipo de estudo, podendo estas estar associadas a Portugal. A espécie de pinheiro utilizada neste estudo foi diferente das que são usualmente utilizadas no Japão e na China. Foi a primeira vez que foram isoladas e identificadas bactérias de um isolado não virulento do nemátode da madeira do pinheiro.For a long time it was thought that Bursaphelenchus xylophilus was the only agent of the pine wilt disease. Recently, it was discovered that there are bacteria associated with the nematodes that contribute to the pathogenesis of this disease, mainly through the release of toxins that promote the death of the pines. Among the species most commonly found, are bacteria belonging to the Bacillus, Pantoea, Pseudomonas and Xanthomonas genera. The main objective of this work was to study the effect of inoculation of maritime pine (Pinus pinaster) with four different nematode isolates, in the bacterial population of nematodes and trees, at different stages of disease progression. The monitoring of progression of disease symptoms was also recorded. Also, the identification of bacteria isolated from the xylem of trees and the surface of nematodes was performed by classical identification methods, by identification system API20E and by sequencing of bacterial DNA. The results showed that for the symptoms progression, the most striking difference was observed for the pines inoculated with the avirulent isolate, C14-5, which led to a slower and less severe aggravation of symptoms than in pines inoculated with the virulent isolates. It was found that at an earlier stage of the disease, in general, bacterial population of inoculated twigs was lower than what was observed 7 and 14 days after inoculation. In one of the quantification methods more bacteria were recovered from the inoculated pines than from the control pines on the 7th and 14th days after inoculation. A bigger bacterial quantity was isolated from pines inoculated with the nematode isolates HF and 20, and, comparatively, few bacteria were isolated from pines inoculated with the avirulent isolate. The identification system API20E proved to be insufficient and poorly discriminatory in the identification of bacterial species; Enterobacter cloacae species was identified in 79% of the isolated bacterial colonies and seven of these colonies couldn’t be identified by this method. Thus, the adoption of identification molecular methods, through bacterial DNA sequencing, allowed a more reliable identification: eleven different bacterial species within the Bacillus, Citrobacter, Enterobacter, Escherichia, Klebsiella, Paenibacillus, Pantoea and Terribacillus genera were identified. General bacterial diversity increased with the progression of the disease. Bacillus spp. species were predominant at the earlier stage of disease progression and Klebsiella oxytoca species at the later stages. Bacterial species isolated from the surface of nematodes were similar to those isolated from the xylem of pines. In the present work new bacterial species were identified which have never been reported before in this type of study and may be associated with Portugal. P. pinaster, the pine species used in this study, was different from those commonly grown in Japan and China. Furthermore, it was the first time that bacteria were isolated and identified from an avirulent pine wood nematode isolate

Conectividade definida por software em ambientes móveis

Broadband Internet access on trains has become an expected service for passengers, and the rise in quality demand has been posing a challenge to service providers. There are solutions for Internet access on trains that lack the flexibility and redundancy needed for an improved QoS in the network. Thus, this dissertation studies two new load balancing solutions, one distributed and the other centralized. In an emulated train network, routers of each car communicate their network statistics to other nodes of the network, to be used in load balancing decisions. In the distributed solution, each router has the capacity to make load balancing decisions, while in the centralized solution, an onboard controller makes those decisions. The latter solution is based on load balancing in an SDN environment. In this system, a remote entity aggregates the passengers’ traffic and forwards it to an external network, or to the Internet. The systems must provide an improved quality of experience to the passengers on the train and the total avaliable bandwidth should be distributed evenly by all cars. Requirements were established for the systems, which were developed after an in-depth analysis of solutions for Internet access on board found in the literature. After obtaining the results of the performance tests, it was concluded that the systems improve the QoE of the passengers, as long as the network conditions are approximately constant for an extended period.O acesso à Internet de banda larga em comboios tornou-se num serviço esperado por parte dos passageiros e o aumento na exigência de qualidade tem constituído um desafio para os prestadores de serviços. Existem soluções de acesso à Internet em comboios que carecem da flexibilidade e redundância necessárias para uma melhor qualidade de serviço na rede. Assim, esta dissertação estuda duas novas soluções de balanceamento de carga, uma distribuída e outra centralizada. Numa rede de comboio emulada, routers de cada carruagem comunicam as suas estatísticas de rede para outros nós da rede, para posteriormente serem tomadas decisões de balanceamento de carga. Na solução distribuída, cada router tem capacidade de tomar decisões de balanceamento de carga, enquanto que na solução centralizada, um controlador dentro do comboio toma essas decisões. Esta última solução baseiase num balanceamento de carga em ambiente SDN. Neste sistema, uma entidade remota agrega o tráfego dos passageiros e encaminha-o para uma rede externa, ou para a Internet. Os sistemas devem proporcionar uma qualidade de experiência melhorada aos passageiros do comboio e a largura de banda total disponível deve ser distribuída mais uniformememente por todas as carruagens. Requisitos foram estabelecidos para os sistemas, que foram desenvolvidos após uma análise aprofundada das soluções de acesso à Internet em comboios encontradas na literatura. Após a obtenção dos resultados dos testes de desempenho, concluiu-se que as duas soluções melhoram o QoE dos passageiros, desde que as condições da rede sejam aproximadamente constantes durante um certo período.Mestrado em Engenharia de Computadores e Telemátic

Utilization of plant growth-promoting bacteria to ameliorate iron nutrition in legumes

Iron (Fe) deficiency is an important micronutrient disorder that leads to severe yield losses and low nutritional crop value, particularly in calcareous soils. There is an urgent need to find sustainable and greener agricultural practices to achieve higher crop yields with higher nutritional value. Biofortification allows the increase of micronutrient concentrations in edible crop tissues and contributes to achieving such demands. The utilization of bioinoculants (BIs) with plant growth-promoting bacteria (PGPB) has been suggested as a promising approach for biofortification and prevention of Fe deficiency. So far, little work has been done on the role of PGPB in soybean (Glycine max) grown under alkaline conditions. The main purpose of this study was to test the potential of PGPB on plant growth and Fe uptake, unveiling mechanisms underlying Fe uptake and accumulation. Soybean was selected as a model species since it is severely affected by Fe deficiency and several underlying factors related to Fe homeostasis are identified. Firstly, an in-depth and critical literature review was conducted concerning the global importance of Fe and the Fe deficiency, the importance of soybean in the fulfillment of global policies, and the potential of PGPB as a sustainable approach to improve Fe nutrition and cope with Fe deficiency. Concerning the experimental work, the first study of this thesis aimed to evaluate the ability of 24 PGPB strains from a CBQF collection to enhance Fe uptake-related processes in soybean grown for 21 days in calcareous soil. Sphingobium fuliginis ZR 1-6 and Pseudomonas jessenii ZR 3-8 were selected based on their in vitro ability to produce indole-3-acetic acid (IAA), 1-aminocyclopropane-1- carboxylic acid (ACC) deaminase, siderophores, and organic acids, to tolerate high pH, and to reduce Fe3+. Bacterial isolates were inoculated singly and as a mixture, and a series of morphological, physiological, and molecular parameters were evaluated. S. fuliginis improved ferric chelate reductase (FC-R) activity (111 %), FRO2 expression (646 %), and root Fe (62 %); combined inoculation fostered Fe accumulation in trifoliates (144 %) and increased IRT1 (239 %) and FER4 expression (5036 %). Overall, S. fuliginis alone or in combination with P. jessenni were the best treatments. In a second study, PGPB were isolated from root tissues and rhizosphere of soybean grown in a Portuguese soil; 76 bacterial strains were isolated from roots (53 %), rhizosphere (29 %), and shoots (18 %), and 29 genera were identified. Two bacterial strains – B. licheniformis P2.3 and B. aerius S2.14 – were selected for in vivo experiments, and inoculated plants were grown to maturity. Photosynthetic parameters, chlorophyll content, total fresh weight, and Fe concentrations were not significantly affected by inoculation. Nevertheless, inoculation with B. licheniformis increased pod number (33 %), decreased FC-R activity (45 %), and increased expression of Fe-related genes; inoculation with B. aerius decreased root length (20 %), FC-R activity (55 %), and FRO2 expression, and increased expression of the remaining genes. Furthermore, inoculation with bacterial isolates improved the accumulation of Mn, Zn, and Ca in soybean tissues. In this study, B. licheniformis showed potential to be incorporated in formulations for improving soybean grown in calcareous soil. The formulation of BIs contemplate a series of requirements and their effective implementation is still challenging. However, they are a promising trend to the accomplishment of future global politics and present a series of advantages to greener agriculture practices that are critically reviewed in the last part of this thesis. In general, the results presented in this thesis contribute to better understand the mechanisms by which PGPB improve Fe uptake and plant growth, under alkaline conditions, and their potential as bioinoculants in a sustainable perspective.A deficiência de ferro (Fe) é uma carência nutricional que leva a graves perdas de produtividade e valor nutricional das culturas, particularmente em solos calcários. Existe uma necessidade urgente de encontrar práticas agrícolas mais sustentáveis para a produção de culturas com maior rendimento e valor nutricional. A biofortificação permite aumentar a concentração de micronutrientes nas culturas e contribui para cumprir estas necessidades. A utilização de bioinoculantes (BIs) com bactérias promotoras de crescimento de plantas (BPCP) tem sido sugerida como uma abordagem de biofortificação e prevenção da deficiência de Fe. Até agora, existem poucos estudos sobre o papel das BPCP em soja crescida em condições alcalinas. O objetivo principal deste estudo foi testar o potencial de BPCP no crescimento das plantas e na absorção do Fe, elucidando os mecanismos subjacentes à absorção e acumulação do Fe. A soja foi escolhida pois é bastante afetada pela deficiência de Fe, e foram identificados vários fatores subjacentes relacionados com a sua homeostasia. Inicialmente, fez-se uma revisão crítica da literatura tendo em conta a importância do Fe e da sua carência, o papel da soja face às políticas globais, e o potencial das BPCP como abordagem sustentável para melhorar a nutrição de Fe e combater a sua deficiência. O primeiro estudo experimental teve como objetivo avaliar a capacidade de 24 estirpes de BPCP de uma coleção do CBQF em melhorar os processos de absorção do Fe em soja crescida em solo calcário durante 21 dias. Sphingobium fuliginis ZR 1-6 e Pseudomonas jessenii ZR 3-8 foram selecionadas com base na sua capacidade in vitro de produzir ácido indol-3-acético (AIA), ácido 1- carboxílico-1-aminociclopropano (ACC) oxidase, sideróforos e ácidos orgânicos, tolerar pH elevado, e reduzir Fe3+. Os isolados bacterianos foram inoculados isoladamente e em mistura, e foram avaliados vários parâmetros morfológicos, fisiológicos e moleculares. S. fuliginis melhorou a atividade da redutase férrica (FC-R) (111 %), a expressão de FRO2 (646 %), e a concentração de Fe na raiz (62 %); a inoculação combinada promoveu a acumulação de Fe nos trifoliados (144 %) e aumentou a expressão de IRT1 (239 %) e FER4 (5036 %). No geral, a inoculação com S. fuliginis sozinho ou em combinação com P. jessenni revelaram-se os melhores tratamentos. Num segundo estudo, as BPCP foram isoladas dos tecidos e da rizosfera de soja cultivada num solo português; foram isoladas 76 estirpes das raízes (53 %), rizosfera (29 %), e parte aérea (18 %), e foram identificados 29 géneros bacterianos. Foram selecionadas duas estirpes – B. licheniformis P2.3 e B. aerius S2.14 – para experiências in vivo em plantas crescidas até à maturidade. Os parâmetros fotossintéticos, teor de clorofila, peso fresco total e concentrações de Fe não foram afetados significativamente pela inoculação. No entanto, a inoculação com B. licheniformis aumentou o número de vagens (33 %), diminuiu a atividade da FC-R (45 %) e aumentou a expressão de genes relacionados com o Fe; a inoculação com B. aerius diminui o comprimento da raiz (20 %), a atividade da FC-R (55 %) e a expressão de FRO2, e aumentou a expressão dos restantes genes. Além disso, a inoculação com as bactérias melhorou a acumulação de Mn, Zn e Ca nos tecidos da soja. Neste estudo, B. licheniformis apresentou potencial para ser incorporado em formulações para o melhoramento da soja crescida em solo calcário. A formulação de BIs contempla vários requisitos e sua implementação representa ainda um desafio. No entanto, estes são uma tendência promissora para o cumprimento das políticas globais futuras, apresentado várias vantagens para práticas agrícolas mais “verdes”, analisadas na última parte desta tese. No geral, os resultados apresentados contribuem para compreender melhor os mecanismos pelos quais as BPCP melhoram a absorção do Fe e o crescimento das plantas em condições alcalinas, e demonstram o seu potencial como bioinoculantes numa perspetiva sustentável

Psychological distress symptoms and resilience assets in adolescents in residential care

Most studies with institutionalised children and adolescents focus on evaluating the impact of negative life events on emotional development. However, few have investigated the relationship between resilience assets and the teenagers’ psychopathological problems. The purpose of the present study was to investigate differences in psychological distress symptoms and in resilience assets in institutionalised and non-institutionalised adolescents. A total of 266 adolescents aged between 12 and 19 years old took part in the study (60.5% female): 125 lived in residential care and 144 resided with their families. Results found a significant and inverse relation between psychopathology and the perception of individual resilience assets, specifically with self-efficacy and self-awareness in the community sample, and with empathy in the institutionalised sample. Overall, and regardless of the age group, adolescents in residential care tend to perceive themselves as significantly less resilient in perceived self-efficacy and empathy, and they report fewer goals and aspirations for the future. The importance of promoting mental health and resilience assets in adolescents, particularly in those in residential care, is discussed. This can be achieved through early interventions that may prevent emotional suffering and deviant life paths, with transgenerational repercussions.info:eu-repo/semantics/publishedVersio

The shaping of Portuguese Art History in Modern Times

UID/PAM/00417/2019 PTDC/ART-HIS/29837/2017Portuguese art history experienced remarkable development after World War II, especially with the work of José-Augusto França, who was responsible for establishing a historiographic canon for nineteenth- and twentieth-century Portuguese art that still endures. José-Augusto França developed a narrative that held Paris up as an artistic and cultural role model in relation to which he diagnosed a permanent delay in Portuguese art. This essay analyses França’s idea of belatedness in the context of Portuguese art historiography and political history and how it is part of a genealogy of intellectual thought produced in an imperial context, revisiting previous art historians and important authors, such as Antero de Quental and António Sérgio. Moreover, it aims to address how the concept of belatedness was associated with the idea of “civilisation” and the idea of “art as civilisation.” Belatedness also has implications in the constraints and specificities of writing a master narrative in a peripheral country – a need particularly felt in the second half of the twentieth century, to mark a political standpoint against the dictatorship that ruled from 1926 to 1974. Part of the reaction to fascism expressed the desire to follow other nations’ democratic example, but the self-deprecating judgements on Portuguese art were frequently associated with the identification of essentialist motifs – the “nature” of the Portuguese people, their way of thinking, of living, their lack of capacities or skills – and of a self-image of being “primitive” in comparison with other European countries that has antecedents going back to the eighteenth century. I will address the nostalgia for the empire and the prevailing notion of belatedness throughout the twentieth century regarding unsolved issues with that nostalgia.publishersversionpublishe

Operational Risk in centralized Risk Management processes in financial institutions: Case study

Dissertation presented as the partial requirement for obtaining a Master's degree in Statistics and Information Management, specialization in Risk Analysis and ManagementOne of the main reasons for the 2008 financial crisis was the failure in operational risk management. Making this one of the reason why operation risk is having more and more a bigger importance. While considering whether or not to opt for a centralized risk management processes we should be aware of the inherent operational risk coming from it. A financial institution who choose this system must ensure that had analyse and review the potentially negative impact and risks that could came from the choice made, so it can protect the company’s interests. This study aims to analyse the operational risk inherent to the centralized risk management processes in financial institutions through analysing the potential risks associated to the process. The study purpose also to find out the benefits and risks to centralization as well as how to mitigate them. The objective is to analyse whether a centralization of risk management processes brought benefits to a financial institution and what are the operational risks associated to that. This study will be relevant not only in the present by knowing if that was a good decision and add value to the company, but as well it will be helping in the identification, measuring and mitigation of operational risks in the centralization of risk management processes from now on

Program Catapults: education for development in Bairro Padre Cruz

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Economics from the NOVA – School of Business and EconomicsThe aim of this Work Project, in partnership with WACT, is to propose a pilot program based on solid research to extend WACT’s activities in Bairro Padre Cruz, a neighborhood in disadvantaged socio-economic circumstances. Catapultas focuses on children and their education, given its large benefits for current and future generations. Researchers agree that family background is a main driver of educational inequality and its persistence. Catapultas is a mentoring pilot program for 20 children in the 5th grade, it aims to introduce an external element, the mentor, in their lives, leading to improvements in educational and social development

Contribution for physical characterization of aromatic rice

Rice is consumed mainly as whole grain, and quali- ty considerations are much more important than for any other food crop. Rice grain quality preference varies from country to country and among regions. Nowadays, aromatic rice varieties are playing a vital role in global rice trading, and also in Portugal. The economic value of rice depends on its cooking and processing quality, such us water uptake ratio, cooking time and texture properties. Three types of aromatic rice were collected and ana- lyzed for biometry characteristics, cooking time, wa- ter absorption, and texture properties. The rice grains were collected from the Portuguese trade market. Biometric characteristics of all rice grains were eval- uated by S21 (LKL) and C-300 (Kett) colorimeter. The rice flour gels texture was performed by a TPA (texture profile analysis), giving information about adhesive- ness, chewiness, gumminess, hardness, resilience and cohesiveness. The extrusion force was also determined according to ISO 11747:2012. Rice samples are commercially classified as long grains B type, because they presented a length higher than 6 mm and the ratio length/ width higher than 3. The samples presented a high degree of whiteness, with a strictly relationship between the total and vitrea white- ness (r 2 = 0.95). The cooking time varied from 12 to 17 minutes and these properties was strongly related with water uptake (from 155.7 - 209.1 g). Generally, aromat- ic rice cultivars presented different textural properties. Aromatic rice samples are different for hardness, adhe- siveness, gumminess, resilience, and extrusion force. From the results obtained it was concluded that the studied aromatic rice cultivars presented different physical properties, mainly the cooking time, water uptake and texture. These differences could be a com- mercial advantage considering the consumer point of view, because the enterprise could provide specif- ic aromatic rice in order to attend different consumer targets

Biópsia do Gânglio Sentinela em Melanomas Espessos: Estudo Clínico Retrospectivo num Único Centro

Background: Sentinel lymph node biopsy is widely accepted in the staging of melanoma. The prognostic value of this technique seems to be greater in intermediate-thickness melanoma (1-4 mm) than in thick melanoma (>4 mm).Objective: To assess the prognostic value of sentinel node status in patients with thick melanoma.Patients and Methods: The disease-free survival and the overall survival were estimated using Kaplan-Meier curves and a Cox regression model in a sample of patients with thick melanoma.Results: Forty-three patients were included (52.2% male) with a mean age of 63.9 years. Sentinel node biopsy was positive in 20 patients (46.5%). Mean follow-up was 40 months. Patients without sentinel node metastases had a 5-year disease-free survival rate significantly higher than those with positive sentinel node (63% versus 19%, p < 0.05). The 5-year overall survival rate was tendentiously lower in patients with positive sentinel node (52% versus 79%), lacking statistical significance.Conclusion: The sentinel node status was not able to predict the overall survival in our series of thick melanomas, probably due to the high risk of hematic spread. However, sentinel lymph node biopsy provided important prognostic information, since the sentinel lymph node status influenced the disease-free survival.Introdução: A biopsia do gânglio sentinela é um procedimento genericamente aceite no estdiamento do melanoma. O valor prognóstico desta técnica parece ser mais relevante nos melanomas de espessura intermédia (1-4mm) do que nos melanomas espessos (>4mm).Objectivo: Avaliar o valor prognóstico do estado do gânglio sentinela em doentes com melanoma espesso.Doentes e Métodos: A sobrevivência livre de doença e a sobrevivência global foram estimadas pelo método de Kaplan-Meier e por um modelo de regressão de Cox numa amostra de doentes com melanoma espesso.Resultados: Foram incluídos 43 doentes (52.2% do sexo masculino) com uma média etária de 63,9 anos. A biopsia do gânglio sentinela foi positiva em 20 casos (46,5%). O tempo médio de seguimento foi de 40 meses. Os doentes em que não foram encontradas metástases no gânglio sentinela tiveram uma sobrevivência livre de doença aos 5 anos significativamente maior do que o grupo de doentes com gânglio sentinela positivo (63% versus 19%, p<0.05). A sobrevivência global aos 5 anos foi tendencialmente mais baixa em doentes com gânglio sentinela positivo (52% versus 79%), embora sem significado estatístico.Conclusão: O estado do gânglio sentinela não teve influência estatística na sobrevivência global dos doentes da nossa série de melanomas espessos, provavelmente devido ao risco elevado de metastização por via hemática. Porém, a biopsia do gânglio sentinela forneceu informação prognóstica relevante, uma vez que teve clara influência na sobrevivência livre de doença