Inicialmente, os cultivos foram desenvolvidos empiricamente e voltados para a produção de alimento humano. Mais tarde, com a descoberta da utilidade dos ficocolóides, os cultivos passaram a ser realizados também para a produção de biomassa para sua extração. Entretanto, sustentabilidade da indústria de macroalgas reside em grande parte nos cultivos, uma vez que os bancos naturais não são suficientes para atender a demanda crescente. Apesar do ágar estar presente nas paredes celulares de espécies de Gracilaria, seu ágar não era explorado comercialmente por apresentar características consideradas inadequadas pela indústria. A descoberta de que a hidrólise alcalina dos grupos sulfato do ágar aumentaria sua força de gel impulsionou a exploração comercial deste gênero. Assim, espécies pertencentes ao gênero Gracilaria são cada vez mais empregadas para a produção de ágar alimentício e a sua tem sido consideravelmente aumentada por meio do desenvolvimento de técnicas de cultivo. Embora a explotação de macroalgas no Brasil tenha se iniciado por volta de 1940, seu impacto social e econômico é reduzido. Estudos sobre a viabilidade de cultivo de macroalgas foram realizados ao longo da costa brasileira, com resultados positivos sobre o potencial de algumas espécies. Apesar disso, o país ainda não possui cultivos de macroalgas em escala comercial. Com base nestes antecedentes, o presente trabalho avaliou o desempenho do cultivo no mar de Gracilaria domingensis e de G. caudata e caracterizou as propriedades do seu ágar (rendimento e qualidade), na Enseada de Armação do Itapocoroy (Penha, Santa Catarina). Os resultados mostraram que o sistema de cultivo testado para ambas espécies é viável na Enseada. O ágar de G. domingensis extraído com CaCl2 apresentou melhores rendimentos e teores de 3,6-anidrogalactose, enquanto a extração com NaOH mostrou ser a mais adequada para G. caudata e ambas espécies mostraram potencial como matéria-prima para extração de ágar alimentício. / Seaweed cultivation in the world began with empirical methodologies to propagate some species utilized as food. Later on, with the discovery of a process to extract and purify agar, it was soon realized that, because of the large volumes needed by a growing industry this activity would only be sustainable if based on mariculture once the natural beds were being depleted. Despite it was known that agar could also be extracted from Gracilaria species, besides the traditional species of Gelidium, that genus yielded a product with lower value. It was only after the discovery that an alkaline treatment could remove part of the sulphate that reduced the gel strength, and therefore improved the agar quality, that the commercial cultivation of Gracilaria species really got momentum and developed continuously. Nowadays, most of the agar produced in the world is based on Gracilaria spp. Although the exploitation of seaweeds for agar production in Brazil started as early as 1940, up to now our production is still very modest due to the limited biomass in the natural beds. Several attempts to cultivate local Gracilaria spp. have been made, some of which with promising results, but a real commercial mariculture never developed in Brazil so far. Based on that we developed this project aiming at the development of viable techniques to cultivate two species of Gracilaria common at the Enseada de Armação do Itapocoroy (Penha, Santa Catarina): Gracilaria domingensis and G. caudata. We also tested different protocols to better extract the agar from the selected species, comparing their yields and quality. Our results show that with some adaptations of the methodologies for cultivation and agar extraction utilized elsewhere it may be possible to make this a viable alternative.

As proteínas inibidoras de poligalacturonases (PGIPs) presentes na parede celular são capazes de limitar o potencial destrutivo da poligalacturonase (PG) fúngica e, assim, constituem um tipo importante dentre os diversos sistemas de defesa do tecido vegetal frente à infecção fúngica. No mamão, o ataque fitopatogênico é o principal causador de danos pós-colheita, e sua alta susceptibilidade pode estar relacionada com a baixa eficácia ou pouca abundância dos meios de defesa anti-fitopatogênica. Uma vez que isso pode estar relacionado com as PGIPs e nada se conhece sobre o papel dessas proteínas nesse fruto, o objetivo do trabalho foi clonar os genes das PGIPs de mamoeiro e definir seu padrão de expressão em diferentes órgãos e tecidos e ao longo do amadurecimento. Para tanto, foram identificadas no genoma do mamoeiro, a partir de critérios que definem a identidade de uma PGIP, duas prováveis sequências dentre 13 candidatas iniciais. Ambas foram clonadas a partir das sequências genômicas e de cDNA, sequenciadas e sua identidade confirmada, sendo denominadas Cppgip4 e Cppgip6. As análises de expressão relativa em diversos tecidos e idades fisiológicas do mamoeiro demonstraram que os dois genes apresentaram diminuição da expressão com o desenvolvimento dos frutos, sendo que com a polpa apresentou redução dos níveis de expressão relativa de Cppgip4 em até 18 vezes dos 30 dias pós-antese (DPA) ao 9 dias pós-colheita (DPC). Na casca também houve redução significativa da expressão com o desenvolvimento. Para a expressão absoluta, nos frutos, sementes, caules, raízes e folhas, o número de cópias de ambos os transcritos decresceu com o desenvolvimento, sendo cerca de cem mil vezes mais abundante para Cppgip6 que para Cppgip4. As tentativas de expressão de proteínas recombinantes em Pichia pastoris não geraram resultado positivo, provavelmente em virtude das condições ideais de indução ainda não terem sido estabelecidas corretamente para o ensaio. A atividade de PGIPs extraídas diretamente do tecido foi medida por análise de difusão em ágar empregando pectinase de Aspergillus niger e revelou uma tendência à diminuição da porcentagem de inibição à medida que os frutos se desenvolveram, em concordância com os resultados da análise por qPCR. O conjunto de resultados sugere que a expressão varia com o estádio de desenvolvimento do fruto e é tecido-específica, possivelmente em resposta à diferente susceptibilidade dos tecidos ao ataque fitopatogênico, indicando que menores níveis de transcritos e atividade no amadurecimento, período de maior susceptibilidade, poderiam sinalizar para a regulação do processo degradativo marcando o início da senescência. / Polygalacturonase inhibiting proteins (PGIPs) present in plant cell walls are able to inhibit the destructive action of fungal polygalacturonase (PG). In this way, they constitute an important type of plant defense system against fungal infections. In papaya fruit, the pathogenic attack is the main cause of post harvesting loss, and its high susceptibility may be related to the low efficiency or low abundance of anti-phytopathogenic defense. Since this fact could be related to PGIPs expression and little is known about the response of these proteins in the fruit, the aim of the present work was to clone the genes of PGIPs papaya fruit and set their expression pattern in different organs and tissues throughout fruit ripening. Thus, two probable PGIP sequences among 13 initial candidates were identified in the papaya genome by using specific criteria. Both sequences were cloned from cDNA and genomic samples, sequenced and confirmed its identity, and then being named Cppgip4 and Cppgip6. Analysis of relative expression in various tissues at different physiological stages demonstrated that both genes were down regulated during fruit development. The relative expression levels of Cppgip4 in papaya pulp was reduced by 18 times from the 30 days post-anthesis (DPA) to the 9 days post-harvest (DPH). Similarly, gene expression in papaya peel was significant down regulated during fruit development. Absolute expression analysis revealed gene expressions in the fruit pulp, seed, stem, root and leaf were also down regulated within development. Moreover, Cppgip6 gene expression was a hundred thousand times more abundant than Cppgip4. The recombinant protein expression in Pichia pastoris did not result positive, probably because of the ideal conditions of induction have not been properly established the yet. The activity of PGIPs extracted directly from the tissue was measured by the agar diffusion assay using pectinase from Aspergillus niger and showed decrease of inhibition during fruit developed in accordance with the results of the qPCR analysis. Based on the results it is possible to suggest the expression of these genes varies temporally with the developmental stage of the fruit and is tissue-specific, possibly in response to the different susceptibility of tissues to pathogenic attack. In addition, the lowest levels of PGIP expression were achieved at the fruit ripening, when the susceptibility to fungal infection is high and could signal for regulating the degradation process characterized by the onset of senescence.

Las microalgas han demostrado ser la fuente de energía más económica. Además, reduce el dióxido de carbono, también tienen una mayor producción en un corto tiempo y menor espacio a diferencia de otros cultivos. Entre las más de 100 especies de microalgas, el Scenedesmus dimorphus es el que tiene mayor potencial. El objetivo de este trabajo de tesis fue determinar de qué manera con un medio de cultivo para Scenedesmus obliquus var. dimorphus (Turpin) se obtiene cantidad de biomasa y contenido de aceite. Inicialmente, el trabajo experimental consistió en la obtención de la cepa pura de Scenedesmus dimorphus (14 mL). Seguidamente, se realizó el escalamiento de cultivo desarrollándose de la siguiente manera: 100 mL, 250 mL, 500 mL, 1L, 2L y 5L, con el fin de determinar las curvas de crecimiento mediante el conteo celular de S. dimorphus con el uso de la cámara de Neubauer, obteniendo los siguientes resultados: 3’ 451,500 unidades celulares en 120 horas (Bayfolan Forte), 2’ 472,500 unidades celulares en 192 horas (BG-11) y 327,500 unidades celulares en 96 horas (Guillard). Concluyendo que el Bayfolan Forte se obtuvo un mejor resultado. A continuación, se realizó 5 cosechas, luego se usó el método de floculación mediante la utilización del sulfato de aluminio Al2(SO4)3 y posteriormente, para la extracción de aceites, se molió la biomasa seca (36.5 g) seguido por la extracción con disolvente en hexano/ isopropanol, y se obtuvo un total de 30 mL de lípidos. Finalmente, en la parte estadística se comprobó la validez de las hipótesis utilizando diferentes métodos, tales como: Anova, Tukey y T. Student.

<p>Because of their compatibility and precise results bio-optical methods based on measurements of the optical tissue properties gain importance in non-invasive medical therapy and diagnostic. For development and standardization of medical devices optical phantoms are suitable. The present report handles the creation and evaluation of solid tissue phantoms, made up of Agar, Vasolipid and ink utilizing different mixture ratios. After cutting the models in slices of 0.2 to 1.1 mm thickness the absorption- and scattering coefficient were measured using a collimated laser beam setup. As result of the study a formula for the preparation of solid optical tissue phantoms with desired optical properties was found, that is valid for models containing 1.12 % Agar.</p>

Because of their compatibility and precise results bio-optical methods based on measurements of the optical tissue properties gain importance in non-invasive medical therapy and diagnostic. For development and standardization of medical devices optical phantoms are suitable. The present report handles the creation and evaluation of solid tissue phantoms, made up of Agar, Vasolipid and ink utilizing different mixture ratios. After cutting the models in slices of 0.2 to 1.1 mm thickness the absorption- and scattering coefficient were measured using a collimated laser beam setup. As result of the study a formula for the preparation of solid optical tissue phantoms with desired optical properties was found, that is valid for models containing 1.12 % Agar.

photosynthetic bacteria is attractive field as production is fueled by solar energy. Hydrogen production potential of two photosynthetic bacteria R.capsulatus (DSM1710 wild type and R.capsulatus YO3 Hup- uptake hydrogenase deleted mutant strain) were examined in agar immobilized systems. In the present work agar and glutamate concentrations were optimized for immobilization of bacteria while feeding bacteria with 40/2-4 mM acetate/ glutamate. Immobilized bacteria produced hydrogen for 420-1428 hours covering 5-7 rounds. Optimizing of acetate concentration indicated that 60 mM produced the highest observed yield around 90-95%. Results shown that 2.5 mg dry cell weight/mL is the optimum cell concentration for wild type strain while 5 mg dry cell weight/mL was optimum for YO3 strain. Using either glycerol or sodium dithionite caused decrease in hydrogen production capacity of immobilized bacteria. It was observed that agar provided protection against inhibition effect of ammonium. Co- v immobilization of bacteria with packed cells of H. salinarium increased total hydrogen production capacity by about 1.14-1.41 folds. Hydrogen production by immobilized bacteria in panel photobioreactor was achieved by a novel system which allowed long term hydrogen production. Immobilized R. capsulatus DSM 1710 in panel reactor worked for about 67-82 days covering 4-5 rounds while immobilized R. capsulatus YO3 worked for 69-72 days covering seven rounds.

>Magister Scientiae - MSc / Informal settlements are often implicated in surface water pollution with faecal matter. In most instances faecal pollution in the associated surface waters persists despite improvements in sewage removal infrastructure. This study evaluates the importance of investigating the water quality of the Plankenbrug River before it reaches Khayamnandi settlement by comparing water quality in spring and in winter upstream (Pre-Khayamnandi) and downstream (Post- Khayamnandi) from the settlement. In this study, faecal indicator bacteria (Escherichia coli and total coliforms) were enumerated using Chromocult agar. E. coli was further characterized with analytical profiling index (API) and haemolysis assays. Both Pre- and Post-Khayamnandi were not significantly different from each other for both total coliforms and E. coli in winter. Pre-Khayamnandi had between 105 and 108 cfu/100 ml for total coliforms while Post-Khayamnandi had total coliform colony count between 106 and 107 cfu/100 ml. E. coli also exhibited a similar pattern with slightly higher counts at Post-Khayamnandi with colony counts from 104 to 107 and 105 to 107 cfu/100 ml. Spring microbial count demonstrated a significant difference to winter counts within each test site (p ≤ 0.01) and across the two sites (p ≤ 0.05). Both total coliforms and E. coli were 102 fold higher at Post-Khayamnandi than at Pre-Khayamnandi in spring. The API assay demonstrated significant difference (p ≤ 0.05) between the two test sites. Pre- Khayamnandi predominantly had two different profiles while Post-Khayamnandi had three. These profiles represented five distinct E. coli biotypes. Sorbitol and sucrose tests within the API assay demonstrated significant differences (p ≤ 0.05) between the two test sites. The prevalence of sorbitol fermenters at Pre-Khayamnandi was 100% while at Post-Khayamnandi it was 73%. Pre-Khayamnandi also demonstrated a significantly higher prevalence of sucrose fermenters than Post-Khayamnandi at 100% and 59% respectively. These differences indicated dissimilar sources of faecal contamination around these sites. Differences in the distributions of sorbitol and sucrose fermenting biotypes demonstrate different toxicity potentials across these two test sites. The haemolysis assay demonstrated that 9% of isolates were haemolytic with reference to both known α- and β-haemolyitic streptococci at Post-Khayamnandi. At Pre-Khayamnandi there was a higher percentage of α- and β-haemolyitic species, 29% and 28%, respectively. Post- Khayamnandi and Pre-Khayamnandi were significantly different from each other with reference to both α- and β-haemolysis (p ≤ 0.05). These haemolytic activities also demonstrate different toxicity potentials across the two sites. In conclusion Khayamnandi contributes to an already heavy faecal load in the Plankenbrug River. Thus remedial measures to maintain high surface water quality of Plankenbrug River should be directed upstream from the Khayamnandi settlement as well as within the settlement equally. This study recommends integration of microbial loads with programs such as the National Microbial Monitoring Program of South Africa to drive prioritization process in directing reclaiming of water quality, inter alia.

Submitted by Pós graduação Farmácia (ppgfar@ufba.br) on 2017-05-15T19:02:04Z No. of bitstreams: 1 ELIZABETE DE JESUS.pdf: 2250219 bytes, checksum: 4edb78551b0744264cf70e273e7072bc (MD5) / Approved for entry into archive by Patricia Barroso (pbarroso@ufba.br) on 2017-05-23T21:16:42Z (GMT) No. of bitstreams: 1 ELIZABETE DE JESUS.pdf: 2250219 bytes, checksum: 4edb78551b0744264cf70e273e7072bc (MD5) / Made available in DSpace on 2017-05-23T21:16:42Z (GMT). No. of bitstreams: 1 ELIZABETE DE JESUS.pdf: 2250219 bytes, checksum: 4edb78551b0744264cf70e273e7072bc (MD5) / FAPESB / O diagnóstico definitivo da estrongiloidíase é realizado pela pesquisa de larvas nas fezes utilizando o método de Baermann-Moraes, apesar da cultura em placa de agar (CPA) apresentar maior sensibilidade. Na maioria das infecções pelo Strongyloides stercoralis, a carga parasitária é baixa e a eliminação das larvas se faz de maneira intermitente, comprometendo a eficácia e precisão do diagnóstico. A pesquisa de anticorpos circulantes através do imunoensaio (ELISA) possui elevada sensibilidade e é utilizado também no auxílio ao diagnóstico da estrongiloidíase. Os objetivos deste trabalho foram (1) comparar os métodos de CPA, Baermann-Moraes e sedimentação espontânea para o diagnóstico da estrongiloidíase (2) avaliar o padrão de migração das larvas de S. stercoralis e ancilostomídeos em placas de agar e o tempo para positividade das culturas, (3) avaliar a interferência da refrigeração das fezes na viabilidade das larvas e (4) padronizar o ELISA para pesquisa de anticorpos anti-S. stercoralis. Para avaliar as sensibilidades dos métodos parasitológicos foram utilizadas 725 amostras de fezes de pacientes atendidos no Laboratório de Análises Clínicas da Faculdade de Farmácia, UFBA, e amostras de soros de 50 pacientes com S. stercoralis, 80 de indivíduos com outras parasitoses e 60 soros controles negativos. A CPA mostrou maior sensibilidade tanto para Strongyloides stercoralis (95%) como para os ancilostomídeos (77%). As sensibilidades da CPA e do método de Baermann-Moraes reduziram aproximadamente 50% quando as amostras foram conservadas a 4ºC. Através do padrão de migração das larvas foi possível realizar a diferenciação de 95,7% das amostras positivas para Strongyloides stercoralis e de 96,7% para os ancilostomídeos, confirmadas através da microscopia. O tempo de visualização macroscópica dos caminhos, deixados pelas larvas também variou significativamente (p < 0,05) entre S. stercoralis e ancilostomídeos, ocorrendo principalmente no primeiro e quarto dias de incubação, respectivamente. No ELISA para detecção de IgG a sensibilidade variou de 72 a 76%, e a especificidade foi de 92,8%, não apresentando diferenças significativas quando o antígeno foi tratado ou não com metaperiodato de sódio. Por outro lado, o ELISA para pesquisa de IgE apresentou sensibilidade de 80 % e após o tratamento do antígeno com metaperiodato de sódio reduziu para 74% (p > 0,05). Enquanto não houve variações nas especificidades, obtidas nos ensaios utilizando soros normais como controles negativos. O número de reações cruzadas de soros de pacientes com outras helmintoses aumentou em torno de 16% quando o antígeno foi tratado com o oxidante. Neste estudo, foi observado que houve uma redução nas médias das densidades ópticas dos soros de pacientes infectados com S. stercoralis com relação aos soros normais, quando o antígeno foi tratado com metaperiodato de sódio, demonstrando a importância dos epitopos glicosilados para o reconhecimento dos anticorpos anti-S. stercoralis. A depleção de IgG dos soros falso-negativos elevou a sensibilidade do ELISA-IgE, porém houve um aumento do número de reatividades cruzadas. A CPA é o método parasitológico mais sensível para o diagnóstico da estrongiloidíase e deve ser recomendado no diagnóstico de rotina. O ELISA pode ser utilizado no auxílio diagnóstico, principalmente em casos pacientes de imunossuprimidos ou antes de terapia imunossupressora. / Definitive diagnosis of strongyloidiasis is usually made by detection of larvae in stool samples using the Baermann-Moraes method, although fecal cultures in agar plate (CPA) have shown higher sensitivity. In the majority of Strongyloides stercoralis infections, the parasite load is low and the larvae output is irregular, influencing the effectiveness and accuracy of diagnosis. The detection of circulating antibodies by enzyme-linked immunosorbent assay (ELISA) has high sensitivity and it is also used as a tool for strongiloidiasis diagnosis. The objectives of this work were (1) to compare the methods of CPA, Baermann-Moraes and sedimentation for diagnosis of strongyloidiasis, (2) to evaluate the migration pattern of the S. stercoralis and hookworm larvae and the time for positivity of cultures, (3) to evaluate the influence of stool refrigeration on the viability of larvae and (4) to standardize the ELISA protocol to dilution serum anti-S. stercoralis IgG antibodies. To determine the sensitivity of the diagnosis methods, 725 stool samples from patients attending to the Clinical Laboratory Analysis of Faculty of Pharmacy, UFBA and serum samples from 50 patients with S. stercoralis, 80 with other parasitic infections and 60 negative control sera were used. The CPA was the most sensitive for both S. stercoralis (95%) and for hookworm (77%). The sensitivities of CPA and Baermann-Moraes decreased about 50% when samples were stored at 4º C. The migration pattern of larvae permitted the differentiation of 92 (95.7%) S. stercoralis and of 89 (96.7%) hookworm positive samples, confirmed by microscopy. The time for visible tracks on agar varied significantly (P < 0.05) between S. stercoralis and hookworms, occurring mainly in the first and fourth day of incubation, respectively. The sensitivity of ELISA for IgG anti-S. stercoralis range from 72% to 76%, and specificity was 91.7%, not showing significant differences when the antigen was treated or not with sodium metaperiodate. On the other hand, the ELISA for IgE had a sensitivity of 80% and after treatment of the antigen with sodium metaperiodate it significantly decreased to 74% (P > 0.05), while the specificities of ELISA-IgE, calculated using normal sera as negative controls, did not present significant variations. However, the number of cross-reactions in ELISA-IgE increased approximately 18% when the antigen was treated with the oxidizing agent. In this study, it was observed a reduction in the mean optical densities of sera from patients infected with S. stercoralis, compared with normal serum, when the antigen was treated with sodium metaperiodate, demonstrating the importance of glycosylated epitopes for the recognition by anti-S. stercoralis antibodies depletion of IgG from false-negative serum, increased the sensitivity of ELISA-IgE, even though there was an increase in cross-reactivity. The CPA is the most sensitive method for the parasitological diagnosis of strongyloidiasis and it should be recommended for routine diagnosis. ELISA should be used for diagnosis of S. stercoralis, especially in cases of immunosuppressed patients or before immunosuppressive therapy.

Made available in DSpace on 2014-10-09T12:25:38Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:02:53Z (GMT). No. of bitstreams: 1 06791.pdf: 4413729 bytes, checksum: 001f9b2a27b3de771b28bba4bb80c4cd (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

Made available in DSpace on 2014-10-09T12:25:38Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:02:53Z (GMT). No. of bitstreams: 1 06791.pdf: 4413729 bytes, checksum: 001f9b2a27b3de771b28bba4bb80c4cd (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP