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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Monitoring cyanotoxins in the western basin of Lake Erie using emerging technologies

Beecher, Amber A. 11 July 2022 (has links)
No description available.
2

Effects of Chemical Properties of Cyanotoxins on Transport through Granular Activated Carbons

Chen, Bingran 30 October 2018 (has links)
No description available.
3

ValidaÃÃo da metodologia para anÃlise de saxitoxina e dc-saxitoxina em Ãgua via derivatizaÃÃo prÃ-coluna / VALIDATION OF METHODS FOR ANALYSIS OF saxitoxin and dc-saxitoxin WATER VIA pre-column derivatization

Larissa Sousa Silvino 21 May 2014 (has links)
A intensificaÃÃo da eutrofizaÃÃo nos mananciais à provocada pelas aÃÃes antrÃpicas e tem como predominÃncia as floraÃÃes de cianobactÃrias. Por sua vez, estas floraÃÃes alteram a qualidade da Ãgua para o abastecimento da populaÃÃo, e, ao serem lisadas, podem liberar toxinas (cianotoxinas) causando intoxicaÃÃo. A primeira confirmaÃÃo in loco da morte de seres humanos por intoxicaÃÃo com cianotoxinas no Brasil levou à OMS a publicar rapidamente novas Portarias sobre o monitoramento da qualidade da Ãgua bruta que incorporaram novos indicadores, como a concentraÃÃo das cianobactÃrias e de suas toxinas nos mananciais utilizados para abastecimento de Ãgua potÃvel. Com isso, as tecnologias para o tratamento da Ãgua e para a identificaÃÃo e quantificaÃÃo das cianotoxinas vÃm passando por um processo de aperfeiÃoamento. Neste contexto, o presente trabalho objetivou validar o mÃtodo de cromatografia lÃquida de fase reversa com detector de fluorescÃncia (CLAE-FLD) e derivatizaÃÃo prÃ-coluna para detecÃÃo e quantificaÃÃo das cianotoxinas saxitoxina (STX) e decarbamoil-saxitoxina (dc-STX) proveniente do cultivo da cianobactÃria Cylindrospermopsis raciborskii. Esta validaÃÃo foi realizada para dar credibilidade ao mÃtodo analÃtico e os parÃmetros selecionados foram: seletividade, linearidade, limite de detecÃÃo (LD) e quantificaÃÃo (LQ), exatidÃo, precisÃo e robustez. Os resultados obtidos apresentaram boa seletividade, comprovando que o mÃtodo possuÃa capacidade de medir as toxinas em uma matriz PÃs ExtraÃÃo na presenÃa de outros componentes. As curvas analÃticas foram construÃdas com nove pontos a partir dos padrÃes de STX e dc-STX. O mÃtodo apresentou uma linearidade no intervalo de 4,5 à 150 Âg L-1 para STX e 3,0 à 132 Âg L-1 para dc-STX, e o coeficiente de correlaÃÃo (r) maior que 0,99 para as duas toxinas, mostrando que o mÃtodo tem a capacidade de fornecer resultados diretamente proporcionais à concentraÃÃo dos analitos detectados. A sensibilidade foi medida atravÃs do LD e LQ, obtendo resultados satisfatÃrios para os objetivos do trabalho. O mÃtodo obteve boa precisÃo e exatidÃo, visto que para STX e dc-STX os diferentes nÃveis de concentraÃÃo estavam com valores dentro dos intervalos permitidos pelas normas brasileiras de validaÃÃo, e apresentou-se robusto, pois foi insensÃvel a pequenas variaÃÃes possÃveis de ocorrer durante a anÃlise. Em resumo, pode-se considerar que o mÃtodo utilizado para a detecÃÃo e quantificaÃÃo das cianotoxinas STX e dc-STX apresentou resultados satisfatÃrios, uma vez que os parÃmetros analisados para validÃ-lo estavam em conformidade aos valores aceitos nas normas brasileiras. / The intensification of eutrophication in the watershed is caused by human actions and is the predominant cyanobacteria. In turn, these blooms affecting the quality of the water supply for the population, and, when disrupted, can release toxins (cyanotoxins) causing intoxication. The first in situ confirmation of the death by poisoning of humans with cyanotoxins in Brazil led the WHO to quickly publish new Ordinance on monitoring of raw water quality that incorporate new indicators, as the concentration of the cyanobacteria and their toxins in water sources used for drinking water supply. Thus, technologies for water treatment and for the identification and quantification of cyanotoxins have been going through a process of improvement. In this context, this study aimed to validate the method of reverse phase liquid chromatography with fluorescence detection (HPLC-FLD) and pre-column derivatization for detection and quantification of cyanotoxins saxitoxin (STX) and decarbamoil-saxitoxin (dc-STX) from the cyanobacterium Cylindrospermopsis raciborskii cultivation. This validation was performed to give credibility to the analytical method and the selected parameters were: selectivity, linearity, limit of detection (LOD) and quantification (LOQ), accuracy, precision and robustness. The results showed good selectivity, confirming that the method had the ability to measure the toxins in a Post Extraction matrix in the presence of other components. The analytical curves were constructed with nine points from the patterns of STX and dc-STX. The method showed linearity in the range of 4.5 to 150 mg L-1 for STX and 3.0 to 132 mg L-1 to dc-STX, and the correlation coefficient (r) greater than 0.99 for both toxins, showing that the method has the capacity to deliver results directly proportional to the concentration of analyte detected. The sensitivity was measured by the LD and LQ, obtaining satisfactory for the purposes of work results. The method achieved good precision and accuracy, whereas for STX and dc-STX different concentration levels were with values ​​within the ranges allowed by Brazilian standards for validation, and showed to be robust because it was insensitive to small variations possible to occur during analysis. In summary, one can consider that the method used for the detection and quantification of cyanotoxins STX and dc-STX showed satisfactory results, since the parameters analyzed to validate it were in conformity with the accepted values ​​in Brazilian standards.
4

IdentificaÃÃo de cianobactÃrias produtoras de saxitoxinas em reservatÃrio de usos mÃltiplos no semiÃrio cearense / Identification of cyanobacteria producing saxitoxins reservoir multipurpose semiÃrio in CearÃ

Ismael Keslley Carloto Lopes 16 August 2013 (has links)
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / No semi-Ãrido brasileiro, devido Ãs caracterÃsticas climÃticas, ao regime irregular de chuvas e Ãs caracterÃsticas do solo, a principal fonte de Ãgua para abastecimento humano tem sido os reservatÃrios artificiais de superfÃcie, aÃudes. A ocupaÃÃo e exploraÃÃo desordenada dos recursos naturais nas bacias hidrogrÃficas, alÃm da alta evaporaÃÃo e da radiaÃÃo solar, interferem negativamente na qualidade da Ãgua armazenada. Como consequÃncia, esses reservatÃrios apresentam um processo de eutrofizaÃÃo acelerado, levando frequentemente a floraÃÃes de fitoplÃnctons, comunidade esta predominantemente dominada por CianobactÃrias. Devido grande parte desses organismos serem produtores de toxinas, podendo causar uma sÃrie de problemas à populaÃÃo, decidiu-se monitorar a comunidade fitoplanctÃnica do aÃude SÃtios Novos, reservatÃrio de usos mÃltiplos, localizado no municÃpio de Caucaia-CE, de janeiro de 2010 a junho de 2011 investigando a produÃÃo de toxinas pelos organismos dominantes. Identificou-se 41 tÃxons, 19 de cianobactÃrias e 22 de algas. Apesar da predominÃncia de nÃmero de tÃxons de algas, estas representaram menos de 10% dos organismos quantificados, com exceÃÃo de 1 amostragem. Jà as cianobactÃrias estiveram presentes em todas as amostragens, dominando quantitativamente o ambiente aquÃtico. Dentre o grupo de cianobactÃrias, destacaram-se Planktothrix agardhii (PA) e Cylindrospermosis raciborskii (CR). A abundÃncia de PA ao longo de todo o perÃodo de amostragem variou de 0 a 5.83x105 cel.mL-1, o que representou 94% da abundÃncia total do fitoplÃncton. A densidade mÃdia da PA foi 7.54.104 cel.mL-1. A abundÃncia CR no mesmo perÃodo variou de 0 a 3.32x104 cell.mL-1, representando 18,9% do total da comunidade fitoplanctÃnica. A abundÃncia mÃdia de CR foi 8.21.103 cel.mL-1. As espÃcies dominantes foram isoladas, e realizando-se bioensaios em camundongos, teste ELISA e mÃtodo de detecÃÃo por HPLC, observou-se a produÃÃo de STX e dc-STX pela CR e dc-GTX pela PA. Essa problemÃtica evidencia a necessidade da implementaÃÃo de aÃÃes integradas dos ÃrgÃos ambiental, de gestÃo dos recursos hÃdricos e companhia de sanamento estadual visando a prevenÃÃo e remediaÃÃo do processo de degradaÃÃo da qualidade da Ãgua, atuando na ampliaÃÃo do saneamento bÃsico, na educaÃÃo ambiental, na fiscalizaÃÃo dos usos e ocupaÃÃes do solo na bacia hidrogrÃfica mas, principalmente, no controle das atividades de piscicultura na bacia hidrÃulica, que ocorrem de maneira superintensiva. / In Brazilian semi-arid Northeast region due to irregular rainfall regime, climatic and soil characteristics, the main source of human water supply has been artificial surface reservoirs. Disorderly occupation and exploitation of natural resources on those hydrographic basins, alongside high evaporation and solar radiation, promote negative impact on stored water quality. As a consequence, the reservoirs present themselves in a process of accelerated eutrophication, often leading to phytoplankton blooms, predominantly dominated by Cyanobacteria. Due to many of these organisms are toxin-producing, which can cause a sort of problems to the population, it was decided to monitor the phytoplankton community of a multipurpose reservoir, located in the municipality of Caucaia-CE, from January 2010 to June 2011, as well as investigate the production of toxins by the dominant organisms. We identified 41 taxa, 19 of cyanobacteria and 22 of algae. Despite the predominance of algae over the identified taxa, they accounted for less than 10% of the quantified organisms, except for one sample. However cyanobacteria were present in all samples, quantitatively dominating the aquatic environment. Among the group of cyanobacteria, excelled Planktothrix agardhii (PA) and Cylindrospermosis raciborskii (CR). PA abundance along all sampling period ranged from 0 to 5.83x105 cells.mL-1 , which accounted for 94% of the total phytoplankton abundance. The average density of PA was 7.54.104 cells.mL-1 . The CR abundance in the same period ranged from 0 to 3.32x104 cells.mL-1 , accounting for 18,9% of total phytoplankton community. The average abundance of CR was 8.21.103 cells.mL-1. The dominant species were isolated and carrying out mice bioassay , ELISA and HPLC analysis, we observed the production of STX and dc-STX by CR and dc-GTX by PA. This issue highlights the need for the implementation of integrated actions from environmental agencies, water resources management and sanitation State companies towards the prevention and remediation of the degradation of water quality, acting in the expansion of sanitation, environmental education, oversighting of the use and occupation of the watershed lands, but mainly in the control of aquaculture activities in the hydraulic basin, which occur in a super intensive way.
5

Seasonal Nutrient Limitations of Cyanobacteria, Phytoplankton, and Cyanotoxins in Utah Lake

Lawson, Gabriella Marie 22 July 2021 (has links)
Excess nutrients from human activity trigger toxic cyanobacterial and algal blooms, creating expansive hypoxic dead zones in lakes, damaging ecosystems, hurting local economies, undermining food and water security, and directly harming human health. To identify when and where nutrients limit phytoplankton and cyanobacterial growth, and cyanotoxin concentrations across Utah Lake, USA we conducted four in-situ bioassay studies (563 cubitainers or experimental units) that experimentally added N, P or N+P over the spring, early summer, summer, late summer, and fall in lake water from the top 20 cm of the water column. For our purpose, we defined total phytoplankton as all prokaryotic or eukaryotic organisms containing chlorophyll-a. We evaluated changes in chlorophyll-a and phycocyanin concentrations; the abundance of cyanobacterial species and total phytoplankton species or divisions; cyanotoxin concentrations of the microcystin, anatoxin-a, and cylindrospermopsin; DIN, SRP, TP, and TN concentrations; and other water chemistry parameters. We found that the nutrient limitation of cyanobacteria, and to a lesser extent phytoplankton, was influenced by season and space. Cyanobacteria were often co-limited in the spring or early summer, limited by a single nutrient in the summer, and not limited by N or P in the late summer and fall. Alternatively, phytoplankton were co-limited from the summer into the fall in the main body of the lake and either N limited or co-limited continually in Provo Bay. Microcystis, Aphanocapsa, Dolichospermum, Merismopedia, and Aphanizomenon spp., and Aulacoseira and Desmodesmus spp. and two taxonomical categories of algae (i.e., unicellular and colonial green algae) were primarily associated with cyanobacteria and phytoplankton nutrient limitations. Concentrations of the three cyanotoxins demonstrated a seasonal signal and loosely followed the growth of specific cyanobacteria but was not dependent on total cyanobacterial cell density. The DIN and SRP were biologically available in all water and nutrient treatments with nutrient concentrations declining over the incubation period, suggesting that nutrient levels were not oversaturated. Our results offer insights into specific nutrient targets, species, and, and cyanotoxins to consider in the future to manage Utah Lake.
6

Using Rapid Small Scale Column Testing to Evaluate Granular Activated Carbon Adsorption of Cyanotoxins from Drinking Water

Kelley, Thomas M. January 2017 (has links)
No description available.
7

Causes and Consequences of Algal Blooms in the Tidal Fresh James River

Wood, Joseph 25 April 2014 (has links)
This dissertation includes 3 chapters which focus on algal bloom of the tidal fresh James River. The first chapter describes nutrient and light limitation assays performed on algal cultures and draw conclusions about long-term patterns in nutrient limitation by comparing results with a previous study . This chapter also describes the influence of riverine discharge upon nutrient limitation in a point-source dominated estuary. This chapter was published in Estuaries and Coasts (Wood and Bukaveckas 2014). The second chapter presents the first comprehensive assessment of the occurrence of the cyanotoxin Microcystin in water and biota of the James River. Data presented in this chapter show that bivalve grazing declines in the presence of Microcystin in the water. The chapter also describes feeding habits in fish as a predictor for inter-specific differences in Microcystin accumulation in their tissues. The work presented in this chapter was published in Environmental Science & Technology (Wood et al. 2014). The third chapter describes the fate of algal carbon in the James River Estuary and the importance of autochthonous and allochthonous sources of organic matter in supporting production of higher trophic levels. Here I draw upon ecosystem metabolism data (NPP and R), abundance and grazing estimates for primary consumers and estimates of advective losses of chlorophyll and external inputs of nitrogen to place ‘top-down’ effects in the broader context of factors influencing chlorophyll and nitrogen fluxes in the James. . This chapter also describes results from mesocosm experiments used to assess the influences of grazers on chlorophyll, nutrients and Microcystis. This work will be submitted in the summer of 2014 to the journal Ecosystems.
8

Identificação e quantificação de microcistinas por HPLC em reservatórios de água / Identification and Quantification of microcystins through HPLC in water reservoirs

Borges, Renata Maria Cortez 08 August 2008 (has links)
A oferta de água vem se tornando cada vez mais diminuta à medida que a população, a indústria e a agricultura se expandem. A contaminação dos mananciais gerada pelo descarte de efluentes domésticos e industriais leva a eutrofização, processo pela qual grande aporte de nutrientes, particularmente fosfatos, leva ao crescimento excessivo de algas. As cianobactérias são microrganismos procariontes, que vivem nos ambientes mais adversos. A floração dessas algas quando presente em mananciais destinados ao consumo humano gera sérios problemas à saúde humana, pois algumas dessas algas podem gerar toxinas, conhecidas como hepatotoxinas, neurotoxinas e dermatotoxinas, de acordo com sua ação farmacológica. Dentre as hepatotoxinas encontramos a microcistina, um heptapeptídeo cíclico que pode levar à morte em horas ou dias. O objetivo desse estudo foi viabilizar a técnica HPLC, já proposta por outros autores, para quantificar microcistinas-LR em reservatórios de água, em escala empresarial para ser implementada em laboratórios de análise de águas. Para o desenvolvimento da técnica, foram utilizadas amostras de uma lagoa com floração de Microcystis. Para determinar a eficiência da técnica cromatográfica, foram realizados estudos com outro método, através do kit ELISA. Nessa etapa do trabalho, verificou-se que a técnica HPLC é mais sensível e viável para a quantificação das microcistinas. Nos experimentos realizados com a cromatografia líquida, observou-se que a coluna C-18 LiChrosorb (25 cm) 7 Sm utilizada no método, e o solvente metanol apresentaram grande influência nos resultados. À medida que os experimentos foram executados, verificou-se o decréscimo da sensibilidade da coluna. Os resultados foram satisfatórios apenas após a limpeza realizada na coluna, onde os padrões apresentaram uma curva de correlação igual a 0,92. Este fato leva a concluir que as colunas precisam ser renovadas para análises mais sensíveis, como no caso das microcistinas. As amostras extraídas com metanol apresentaram resultados relevantes, isto é, quanto maior a concentração de metanol utilizado na extração, maior a concentração de microcistina-LR obtida nos resultados, concluindo o metanol ser o solvente apropriado para a extração. Por fim, conclui-se que o método desenvolvido é viável, apresentando algumas dificuldades para sua implantação em escala empresarial. / The water supply has been decreasing more and more as the population, industry and agriculture expand. The contamination of the water sources generated by the domestic and industrial effluents discharges leads to the eutrophization, process where the large presence of nutrients, particularly phosphates, causes excessive increase of algae. The cyanobacteria are procaryote microorganisms which live in the most diverse environments. The florescence of the algae when present in sources directed to human consumption generates serious problems to the human health, for some of them may produce toxins known as hepatotoxins, neurotoxins and dermotoxins, according to their pharmacological action. Among the hepatotoxins, the microcystin, a cyclic heptapeptide that can lead to death in hours or days, is found. The objective of this study was to make feasible the use of the HPLC technique, already proposed by other authors, to quantify microcystins-LR in water reservoirs, in enterprise scale to be implemented in water analysis laboratories. In order to develop the technique, samples of water from a pond with Microcystis florescence were utilized. To evaluate the efficiency of the chromatographic technique, studies were performed with another method, through the ELISA kit. In this phase of the work, it was verified that the HPLC technique is the most sensible and viable for the quantification of the microcystins. It was observed, in the experiments performed with the liquid chromatography, that the column C-18 LiChrosorb (25 cm) 7 Sm utilized in the method and the methanol solvent presented great influence in the results. As the experiments were realized, the decrease of the sensibility of the column was verified. The results were satisfactory only after the column being cleaned, when the patterns presented a curve of correlation equal to 0.92. This fact leads to the conclusion that the columns need to be renewed for more sensible analysis, like in the case of the microcystins. The samples extracted with methanol presented relevant results, that is, the greater the concentration of the methanol utilized, the higher the concentration of microcystin-LR obtained in the results, leading to the conclusion that methanol was the solvent adequate to the extraction. Finally, it was concluded that the method developed is feasible, presenting some difficulties concerning its implantation in enterprise scale.
9

Adsorção com carvão ativado granular e degradação biológica para o tratamento avançado de águas de abastecimento: remoção de microcistina em escala laboratorial / Adsorption with granular activated carbon and biological degradation for advanced treatment of drinking water: removal of microcystin in laboratory scale

Ortolan, Andréia Vanessa Stocco 19 May 2016 (has links)
As cianotoxinas, potencialmente prejudiciais à saúde dos homens e animais, são toxinas produzidas pelas cianobactérias e que podem estar presentes em reservatórios utilizados para captação de água para abastecimento, sobretudo em função do agravamento das condições sanitárias das bacias hidrográficas. Diante disso, torna-se fundamental que a água esteja adequada ao consumo humano em sua distribuição. Uma das formas de tratamento para remoção das cianotoxinas que vem sendo estudada é por meio do uso de carvão ativado granular (CAG), cuja eficiência depende de suas características intrínsecas e das condições operacionais. Outra possibilidade para remoção de tais toxinas se dá por meio da utilização de microrganismos capazes de promover sua biodegradação. A presente pesquisa analisou o potencial de remoção de microcistina por meio de adsorção por três diferentes CAGs comerciais (materiais de origem: casca de coco, mineral e osso) e por meio da degradação biológica por dois gêneros de bactérias. Para isso, foi utilizado um extrato de toxina produzido por meio do cultivo de Microcystis aeruginosa (cepa BB005) em meio WC, ao longo de 30 dias, com fotoperíodo de 12 h. Os carvões foram caracterizados de acordo com a massa específica aparente, umidade, pH, teor de cinzas, número de iodo, índice de azul de metileno e área superficial específica. Em seguida, foram realizados ensaios de adsorção com a microcistina (concentração inicial de 100 µg.L-1) para construção de isotermas com tempo de equilíbrio de 1 h. As análises de toxina foram realizadas pelo método ELISA e os dados foram ajustados aos modelos matemáticos de Langmuir e de Freundlich. Os melhores resultados de adsorção foram obtidos a partir do carvão de origem mineral (99% de remoção), que apresentou valores maiores para número de iodo (710 mgI2.g-1), índice de azul de metileno (169 mL.g-1), área superficial específica (911 m2.g-1) e porcentagem de microporos (70%), com melhor ajuste da isoterma obtido pelo modelo de Freundlich (R2 = 0,88). Os ensaios de biodegradação da microcistina com os gêneros Sphingomonas sp. e Brevundimonas sp foram realizados com a toxina esterilizada e não esterilizada (concentração inicial de 75 µg.L-1). Os resultados demonstraram que tais bactérias não foram capazes de degradar a toxina esterilizada. Entretanto, foi observada degradação nos ensaios em que a toxina não estava esterilizada (porcentagem de remoção de 98%), indicando a presença de algum microrganismo e/ou enzima atuante nesse processo, uma vez que o cultivo da cepa para obtenção do extrato não foi realizado sob condições de assepsia. Recomenda-se, para futuras pesquisas, a identificação do responsável pela biodegradação da microcistina, bem como a aplicação do carvão mineral em maior escala (filtro de leito fixo). Deste modo, será possível avaliar a formação de biofilme no leito de carvão, e comparar o desempenho da adsorção e da biodegradação na remoção do poluente-alvo em escala mais próxima à real. / The cyanotoxins, which are potentially harmful to the health of humans and animals, are toxins produced by cyanobacteria. They can be found in reservoirs used for water supply, especially due to the degradation of sanitary conditions within the watersheds. Therefore, it is essential to assure potable water for human consumption in its distribution. The use of granular activated carbon (GAC) has been studied for advanced water treatment and cyanotoxins\' removal. The efficiency of such technology depends on activated carbon intrinsic characteristics and operating conditions. Removing such toxins through biodegration by microrganisms is another possibility. The present study was perfomed in laboratory conditions and analyzed microcystin removal by adsorption by three commercial GACs (source materials: coconut shell, mineral and bone) and through the biological degradation by two genera of bacteria. An extract of toxin was produced by the cultivation of Microcystis aeruginosa (strain BB005), in medium WC throughout 30 days, with a 12 h photoperiod. The activated carbon samples were characterized regarding the apparent density, moisture, pH, ash content, iodine number, methylene blue index and specific surface area. Thereafter, adsorption experiments were conducted with microcystin (initial concentration of 100 µg.L-1) for estimating isotherms considering an equilibrium time of 1 h. The toxin analyses were performed by the ELISA (Enzyme-Linked Immunosorbent Assay) and the data were adjusted to the mathematical models of Langmuir and Freundlich. The best adsorption results were obtained with the mineral carbon (removal percentages of 98%), which also showed the highest values for the iodine number (710 mgI2.g-1), methylene blue index (169 mL.g-1), specific surface area (911 m2.g-1) and percentage of micropores (70%), with best adjustment of the isotherm through the Freundlich model (R2 = 0.88). The biodegradation tests of microcystin with Sphingomonas sp. and Brevundimonas sp. were carried out with the sterile and non-sterile toxin (initial concentration of 75 µg.L-1). The results showed that these bacteria were not able to promote degradation of the toxin when it was sterilized. However, the degradation was observed in those tests in which the toxin was not sterilized (reduction up to 98%), suggesting the presence of a microrganism and/or an enzyme responsible for this process, since the cultivation of the strain for obtaining the extract was not performed under sterile conditions. As for future research, it is recommendedthe identification of the factor responsible for the biodegradation of the microcystin, as well as the study of the use of the activated carbon from mineral source in a larger scale (fixed-bed filter). This would help to reach a scale closer to the real water treatment plants and evaluate the biofilm formation in the carbon bed, allowing the comparison of the performance of both adsorption and biodegradation processes in the removal of the target pollutant.
10

Cianobactérias de ambiente costeiro: filogenia, prospecção gênica e química de moléculas bioativas / Cyanobacteria from coastal environment: phylogeny, gene and chemical prospecting of bioactive molecules

Vaz, Marcelo Gomes Marçal Vieira 05 August 2014 (has links)
O filo Cyanobacteria constitui um grupo filogeneticamente coerente, embora, apresente grande diversidade morfológica, sendo sua sistemática constantemente revisada. Esses micro-organismos são, ainda, alvos de estudos biotecnológicos em razão da produção de toxinas e na busca por substâncias de interesse farmacológico. Dentre as linhagens analisadas neste estudo, sete sequências do gene rRNA 16S foram geradas e avaliadas com sequências previamente obtidas. Ao menos dois grupos podem representar novos gêneros de cianobactérias, sendo que um grupo demonstra ser endêmico de manguezais brasileiros. Os genes de inibidores de proteases, aeuruginosina, cianopeptolina e microviridina, foram detectados e a produção de aeruginosina foi confirmada por LC-MS nos gêneros Cyanobium e Nostoc. Sequências de aminoácidos do precursor de microviridina indicaram a produção de três novas variantes em quinze linhagens de cianobactérias dos gêneros Cyanobium, Synechococcus, Cyanobacterium, Nodosilinea e Nostoc. O potencial genético para produção de cilindrospermopsina (cyrJ) foi confirmado em vinte e seis linhagens. Em cinco linhagens dos gêneros Cyanobium e Nostoc foram encontrados os genes mcyD, mcyE e mcyG, envolvidos na biossíntese de microcistina. A sequência McyG da linhagem Nostoc sp. CENA175 agrupou-se filogeneticamente com outras de linhagens produtoras de microcistina. Os genes sxtA e sxtI, envolvidos na biossíntese de saxitoxina, foram encontrados em nove linhagens dos gêneros Cyanobium, Oxynema, Leptolyngbya, Nodosilinea e Nostoc. A sequência de SxtI da linhagen Leptolyngbya sp. CENA134 apresentou similaridade >= 70 % com proteínas hipotéticas enquanto as de Nostoc sp. CENA159 e Nostoc sp. CENA160 apresentaram similaridade >= 82 % com O-carbamoiltransferase. Na análise filogenética, a sequência de SxtI da linhagem Nostoc sp. 160 agrupou-se com sequências de linhagens produtoras de saxitoxina. Nas análises químicas, a fração 3 do extrato da linhagem Oxynema sp. CENA135 revelou uma substância com características de ácidos graxos poli-insaturados e a fração 2 do extrato da linhagem Nostoc sp. CENA175 apresentou uma estrutura aromática ligada a uma cadeia alifática. Outros três extratos, obtidos das linhagens Cyanobium sp. CENA157, Nodosilinea sp. CENA183 e Nostoc sp. CENA184 mostraram-se promissores quanto à presença de substâncias nitrogenadas. Os ensaios de bioatividade revelaram que 48 % dos extratos metanólicos inibiram o crescimento de ao menos um isolado de bactéria e/ou levedura. Os extratos das linhagens Cyanobium sp. CENA142 e Cyanobacterium sp. CENA169 foram eficientes contra o crescimento de seis bactérias patogênicas. Nos ensaios de inibição de células tumorais, o extrato de DCM da linhagem Cyanobium sp. CENA154 (100 ?gomL-1) inibiu moderadamente culturas de células 3LL. Os extratos etanólicos de Oxynema sp. CENA135 (20 ?gomL-1) e Cyanobium sp. CENA154 (100 ?gomL-1) inibiram as células CT-26. Em ensaios conduzidos com linhagens de células de glioma (U251), câncer de mama (MCF-7) e câncer de pulmão (NCI-H460), o extrato de DCM da linhagem Cyanobium sp. CENA136 inibiu 50 % do crescimento das respectivas células tumorais nas concentrações 7,8; 27,1 e 14,0 ?gomL-1. Desta forma, além de filogeneticamente diversas, as cianobactérias isoladas de ambiente marinho do Estado de São Paulo constituem fonte promissora de inibidores de proteases, cianotoxinas e substância bioativas com ação antibacteriana, antifúngica e antitumoral. / The phylum Cyanobacteria is a phylogenetically coherent group, although presenting great diversity, and its systematic have been constantly reviewed. These microorganisms are also targets of biotechnological studies due to the production of toxins and the search for novel substances of pharmacological interest. Among the strains analyzed in this study, sequences of the 16S rRNA gene were generated for seven and, than, analyzed with sequences previously obtained. At least two groups may represent new cyanobacterial genera, while a group of Cyanobium proves to be endemic of Brazilian mangroves. Genes of the proteases inhibitors, aeuruginosin, cyanopeptolin and microviridin, were detected and the production of aeruginosin was confirmed by LC-MS for Nostoc and Cyanobium. The amino acid sequences of microviridin precursor indicated the production of three new variants in fifteen cyanobacterial strains of the genera Cyanobium, Synechococcus, Cyanobacterium, Nostoc and Nodosilinea. The genetic potential for production of cylindrospermopsin (cyrJ) was confirmed in twenty-six strains. In five strains of the genera Nostoc and Cyanobium the mcyD, mcyE and mcyG genes, which are involved in the microcystin biosynthesis, were found. The McyG sequence of Nostoc sp. CENA175 was phylogenetically grouped with sequences of microcystin-producing strains. The sxtA and sxtI genes, from saxitoxin biosynthesis, were found in nine strains of the genera Cyanobium, Oxynema, Leptolyngbya, Nodosilinea and Nostoc. The SxtI sequence of Leptolyngbya sp. CENA134 showed similarity >= 70 % with hypothetical proteins, while the sequences of Nostoc sp. CENA159 and Nostoc sp. CENA160 showed similarity >= 82% with O-carbamoyltransferase. In the phylogenetic analysis, the SxtI sequence of Nostoc sp. CENA160 grouped with sequences of strains that produce saxitoxin. In chemical analysis, the fraction 3 of the Oxynema sp. CENA135 extract revealed a substance with poly-unsaturated fatty acids characteristics and the fraction 2 of Nostoc sp. CENA175 extract indicated an aromatic structure, attached to an aliphatic chain. Other three extracts obtained from Cyanobium sp. CENA157, Nodosilinea sp. CENA183 and Nostoc sp. CENA184 were promising for the presence of nitrogenous substances. Bioactivity assays revealed that 48 % of the methanolic extracts inhibited the growth of at least one isolate of bacteria and/or yeast. The extracts of Cyanobium sp. CENA142 and Cyanobacterium sp. CENA169 were efficient against six pathogenic bacteria. In the inhibition assays of tumor cells, the DCM extract of Cyanobium sp. CENA154 (100 mgomL-1) moderately inhibited the growth of 3LL cells. Ethanol extracts of Oxynema sp. CENA135 (20 mgomL-1) and Cyanobium sp. CENA154 (100 mgomL-1) were able to inhibit cultures of CT- 26 cells. In tests conducted with glioma cell lines (U251), breast cancer (MCF-7) and lung cancer (NCI-H460), the DCM extract of Cyanobium sp. CENA136 caused 50 % of growth inhibition, respectively, when used at concentrations of 7.8, 27.1 and 14.0 mgomL-1. Thus, besides their phylogenetically diversity, the cyanobacteria strains from marine environment of the São Paulo state are a promising source of protease inhibitors, cyanotoxins and bioactive compounds with antibacterial, antifungal and antitumor activities.

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