Biocaumulação de alumínio em rebentos micropropagados de Plantago sp.

Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2009

Comparação dos métodos aplicados na detecção de bactérias coliformes, Escherichia coli e Enterococcus sp. em águas para fins recreativos

Dissertação mest., Qualidade em Análises, Universidade do Algarve, 2007

Variabilidade genómica de Citrus tristeza virus e transformação genética de Citrus aurantium L. com vista à obtenção de resistência

Tese de dout., Biologia, Faculdade de Engenharia de Recursos Naturais, Univ. do Algarve, 2003

Sobrevivência e crescimento de juvenis de cavalos-marinhos (Hippocampus sp.): caso de estudo: Hippocampus erectus (Perry, 1810)

Dissertação de mest., Ciências, Faculdade de Ciências do Mar e do Ambiente, Univ. do Algarve, 2009

Estudo preliminar do efeito supressor de genes do Citrus tristeza virus (CTV) no silenciamento pós-transcripcional em Nicotiana benthamiana

Dissertação de mest., Engenharia Biológica, Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2011

Exploring an uncommon host-parasite interaction to unveil the uniqueness of Perkinsus sp as a model organism

We report the exploration of some unique metabolic pathways in Perkinsus olseni a marine protist parasite, responsible to significant mortalities in mollusks, especially in bivalves all around the world. In Algarve, south of Portugal carpet shell clam Ruditapes decussatus mortalities can reach up to 70%, causing social and economic losses. The objective of studying those unique pathways, is finding new therapeutic strategies capable of controlling/eliminating P. olseni proliferation in clams. In that sense metabolic pathways, were explored, and drugs affecting these cycles were tested for activity. The first step involved the identification of the genes behind those pathways, the reconstitution of the main steps, and molecular characterization of those genes and later on, the identification of possible targets within the genes studied. Metabolic cycles were screened due to the fact of not being present in host or differ in a critical way, such as the following pathways: shikimate, MEP-­‐ isoprenoids, Leloir cycle for chitin production, purine biosynthesis (unique among protists), the de novo synthesis of folates (absent in metazoa) and some unique genes like, the alternative oxidase (a branch of respiratory chain) and the hypoxia sensor HPH. All those pathways were covered and possible chemical inhibition using therapeutic drugs was tested with positive results. The relation between the common host Ruditapes decussatus and P. olseni was also explored in a dimension not possible some years ago. With the accessibility to second generation sequencers and microarray analysis platforms, genes involved in host defense or parasite virulence and resistance to the host were deciphered, allowing aiming to new targets (mechanisms and pathways), offering new possibilities for the control of Perkinsus in close environments. The thousands of genes, generated by this work, sequenced and analyzed from this commercial valuable clam and for Perkinsus olseni will be an important and value tool for the scientific community, allowing a better understanding of host-­‐parasite interactions, promoting the usage of P. olseni as an emerging model for alveolata parasites.

Molecular characterization of Citrus tristeza virus isolates from Cyprus on the basis of the coat protein gene

Within the context of a program in Cyprus for the control of Citrus tristeza virus (CTV), the coat protein (CP) genes of 12 local isolates of the virus that induced different symptoms on host trees, were compared to those of known isolates. The CP genes were reverse-transcribed (RT) and amplified by polymerase chain reaction (PCR) and the resulting amplicons were cloned and sequenced. Nucleotide sequence analysis revealed no signs of geographic speciation. All the sequences obtained clustered close to those of previously known isolates of worldwide origin that are in five distinct groups. The nucleotide diversity was high compared to that found using a worldwide database of CP gene sequences. These data support the existence of different CTV introductions into Cyprus or an introduction from a location in which CTV is relatively diverse. Some of the isolates induced stem pitting on branches of grapefruit and sweet orange. Such isolates have not been noted often in the Mediterranean basin. They were close in CP sequence to isolate B249 from Venezuela, which induces stem pitting, and are of particular concern for the whole region.

First report of Citrus tristeza virus in the State Union of Serbia and Montenegro.

Citrus production in the State Union of Serbia and Montenegro has a strategic importance to the agricultural sector. Approximately 400,000 trees are now grown in the major citrus producing region, which is the Montenegrin Coastal Region. Satsuma mandarins and lemons grafted on Poncirus trifoliata are the most cultivated varieties. In December 2003, eight samples taken from the coastal region close to the towns of Bar and Ulcinj were analyzed using enzyme- linked immunosorbent assay (ELISA) with SP7 antibodies produced at Universidade do Algarve, Portugal (3). Further analysis was done using immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) targeting the entire coat protein (CP) gene (forward primer CTV1: 5(prime)- ATGGACGACGAAACAAAGAA-3(prime) and reverse primer CTV10: 5 (prime)-ATCAACGTGTGTTGAATTTCC-3(prime)). Using both techniques, seven of eight samples analyzed were found to be infected by Citrus tristeza virus (CTV), including samples from five trees that exhibited chlorosis, gummosis, and fruit deformation, and two trees that were symptomless.

Occurrence of stem-pitting strains of Citrus tristeza virus in Croatia

Citrus is grown in Croatia (approximately 1,500 ha of citrus groves) on the Dalmatian Coast and Islands between 42 and 43°30'N. The major species, Citrus unshiu Marc. (Satsuma mandarin), is grafted on trifoliate rootstock. The presence of Citrus tristeza virus (CTV) in Satsumas in the Neretva Valley Region was previously reported (3). During the course of a biomolecular characterization of isolates from Croatia, 15 budsticks were collected from field- infected, enzyme-linked immunosorbent assay (ELISA)-positive sources during the autumn of 2003 near Kaštela, Split, Metković (Neretva Valley), and on the island of Vis. Isolates were propagated by graft transmission to Madam Vinous sweet orange (SwO) and maintained in an insect-proof greenhouse at 21 to 33° C.

Ascorbic acid and flavanone glycosides in citrus: Relationship with antioxidant activity

The effects of cultivar and orchard on the content of ascorbic acid and flavanone glycosides as well as on the antioxidant activity of citrus juices were studied. Ascorbic acid and flavanone glycosides were determined using HPLC method. For antiradicalar activity evaluation, TAEC and ORAC methods were used. Citrus of 'Valencia late' from Faro and 'D. Joao' presented the highest levels of hesperidin in contrast to the citrus of 'Valencia late' from Silves with 0.4 mg/ml, a value closer to those of Navel oranges 'Navelate' and 'Lanelate'. Citrus of 'Valencia late' from Silves and 'D. Joao' presented the lowest levels of narirutin. 'Ortanique' and 'Encore' cultivars were poorer in ascorbic acid compared with the oranges. When the TEAC method was used for the determination of antioxidant activity, 'Valencia late' from Faro presented the best activity. A good relation was observed between antiradical activity measured through the TEAC method and the amount of ascorbic acids in samples. None relation was found between the levels of flavanones and antiradical activity. When the ORAC method was applied, the best activity was found for 'Navelate' and 'Encore' samples and practically none relation between this method and the amounts of ascorbic acid and flavanones was found.

Caraterização dos supressores p20 e p23 do Citrus tristeza virus e avaliação da possibilidade da sua inibição

No presente trabalho pretendeu-se caracterizar a capacidade supressora das proteínas p20 e p23 de diferentes grupos filogenéticos do CTV e o possível silenciamento da p23 de forma a ser incluída numa estratégia de proteção. A atividade supressora local da proteína p23 de todos os grupos filogenéticos foi caracterizada em Nicotiana benthamiana da linha 16C. Todas as proteínas testadas foram capaz de suprimir o silenciamento local, mas não o silenciamento a curta distância. A supressão local mais eficiente verificou-se para a p23 do Gp 5 e a menos eficiente para os isolados dos Gps 2 e M. Surpreendentemente, a p23 do Gp 5 aboliu completamente o silenciamento sistémico, sugerindo que existe uma relação entre a intensidade do silenciamento local e sistémico. A capacidade supressora local conjunta das proteínas p20 e p23 foi avaliada. A coexpressão de ambas as proteínas revelou atividade supressora mais forte comparada com a capacidade de cada proteína individual, mesmo quando inoculada com metade da densidade ótica, sugerindo a existência de sinergismo entre as proteínas p20 e p23. Para analisar as propriedades supressoras a longo prazo, as proteínas p20 e p23 foram inseridas no vetor viral TRV que assegurou a sua disseminação pela planta e expressão por um período mais alargado. Foram observados sintomas em N. benthamiana para todas as modalidades testadas, tais como, nanismo da planta, lesões necróticas severas nas folhas inoculadas e nas folhas novas ligeiros sintomas de mosaico e enrolamento. Contudo, sistemicamente não foram registadas diferenças na capacidade supressora das proteínas p20 e p23. A possibilidade para silenciar sistemicamente a proteína p23 quando incluída num genoma viral foi avaliada através do uso de plantas e enxertos transgénicos para a p23. A estratégia que envolve o uso de enxertos transgénicos parece indicar resultados promissores que conduzem ao silenciamento da p23, contudo, são resultados que devem ser encarados como preliminares.

Otimização da fragmentação e dos processos de fixação na reprodução assexuada do coral mole Sarcophyton sp.

Dissertação de mestrado, Aquacultura e Pescas (Aquacultura), Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Definição do modo de operação, descontínuo ou semi-descontínuo, para produção de bioetanol utilizando um novo substrato misto pela levedura Kluyveromyces sp.

Dissertação de mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Caracterização funcional da microalga extremófila Coccomyxa sp.

Dissertação de mestrado, Biologia Molecular e Microbiana, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Dynamics of production and mortality of Aurelia SP. in thau lagoon, northwerstern Mediterranean

Dissertação de mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015