Bioremediation of phosphates in seawater: approach for recirculating marine aquaculture effluents

Marine Recirculating Aquaculture Systems (RAS) produce great volume of wastewater, which may be reutilized/recirculated or reutilized after undergoing different treatment/remediation methods, or partly discharged into neighbour water-bodies (DWW). Phosphates, in particular, are usually accumulated at high concentrations in DWW, both because its monitoring is not compulsory for fish production since it is not a limiting parameter, and also because there is no specific treatment so far developed to remove them, especially in what concerns saltwater effluents. As such, this work addresses two main scientific questions. One of them regards the understanding of the actual (bio)remediation methods applied to effluents produced in marine RAS, by identifying their advantages, drawbacks and gaps concerning their exploitation in saltwater effluents. The second one is the development of a new, innovative and efficient method for the treatment of saltwater effluents that potentially fulfil the gaps identified in the conventional treatments. Thereby, the aims of this thesis are: (i) to revise the conventional treatments targeting major contaminants in marine RAS effluents, with a particular focus on the bioremediation approaches already conducted for phosphates; (ii) to characterize and evaluate the potential of oyster-shell waste collected in Ria de Aveiro as a bioremediation agent of phosphates spiked into artificial saltwater, over different influencing factors (e.g., oyster-shell pre-treatment through calcination, particle size, adsorbent concentration). Despite the use of oyster-shells for phosphorous (P) removal has already been applied in freshwater, its biosorptive potential for P in saltwater was never evaluated, as far as I am aware. The results herein generated showed that NOS is mainly composed by carbonates, which are almost completely converted into lime (CaO) after calcination (COS). Such pre-treatment allowed obtaining a more reactive material for P removal, since higher removal percentages and adsorption capacity was observed for COS. Smaller particle size fractions for both NOS and COS samples also increased P removal. Kinetic models showed that NOS adsorption followed, simultaneously, Elovich and Intraparticle Difusion kinetic models, suggesting that P removal is both a diffusional and chemically rate-controlled process. The percentage of P removal by COS was not controlled by Intraparticle Diffusion and the Elovich model was the kinetic model that best fitted phosphate removal. This work demonstrated that waste oyster-shells, either NOS or COS, could be used as an effective biosorbent for P removal from seawater. Thereby, this biomaterial can sustain a cost-effective and eco-friendly bioremediation strategy with potential application in marine RAS.

Síntese de ésteres catalisada por fosfatos e fosfonatos metálicos(IV)

No âmbito do Mestrado em Química, com especialização em Química Orgânica e Produtos Naturais, será apresentado nesta dissertação o trabalho desenvolvido sobre a síntese e caracterização de fosfatos e fosfonatos metálicos tetravalentes, assim como a avaliação da sua capacidade catalítica em reações de esterificação. Iniciou-se este trabalho pela síntese de vários fosfatos e fosfonatos metálicos tetravalentes, que apresentam a fórmula geral MIV(HXO4)2.nH2O e MIV(RXO3)2.nH2O, respetivamente, onde MIV = Zr, X = P, W, Mo e R = grupo orgânico. Os compostos sintetizados foram caracterizados por espetroscopia de infravermelho, difração de raios-X de pós e por análise elementar e termogravimétrica. Os fosfatos e fosfonatos metálicos tetravalentes foram avaliados como catalisadores em reações de esterificação de Fischer entre os ácidos acético, ou benzóico, e diferentes álcoois. Após esta avaliação foram efetuados estudos de otimização das condições reacionais para a síntese de cada éster. Em cada um dos casos foi utilizado o catalisador com o qual se obtiveram os melhores rendimentos nos estudos iniciais, ou seja com o molibdato-fenilfosfonato de zircónio(IV), ZrMoPhP. Todas as reações foram monitorizadas por cromatografia gasosa ou por cromatografia líquida de alta pressão. Todos os ésteres foram identificados por comparação com os padrões correspondentes.

Functional alkali tantalate 2D structures for microelectronics and related applications

Alkali tantalates and niobates, including K(Ta / Nb)O3, Li(Ta / Nb)O3 and Na(Ta / Nb)O3, are a very promising ferroic family of lead-free compounds with perovskite-like structures. Their versatile properties make them potentially interesting for current and future application in microelectronics, photocatalysis, energy and biomedics. Among them potassium tantalate, KTaO3 (KTO), has been raising interest as an alternative for the well-known strontium titanate, SrTiO3 (STO). KTO is a perovskite oxide with a quantum paraelectric behaviour when electrically stimulated and a highly polarizable lattice, giving opportunity to tailor its properties via external or internal stimuli. However problems related with the fabrication of either bulk or 2D nanostructures makes KTO not yet a viable alternative to STO. Within this context and to contribute scientifically to the leverage tantalate based compounds applications, the main goals of this thesis are: i) to produce and characterise thin films of alkali tantalates by chemical solution deposition on rigid Si based substrates, at reduced temperatures to be compatible with Si technology, ii) to fulfil scientific knowledge gaps in these relevant functional materials related to their energetics and ii) to exploit alternative applications for alkali tantalates, as photocatalysis. In what concerns the synthesis attention was given to the understanding of the phase formation in potassium tantalate synthesized via distinct routes, to control the crystallization of desired perovskite structure and to avoid low temperature pyrochlore or K-deficient phases. The phase formation process in alkali tantalates is far from being deeply analysed, as in the case of Pb-containing perovskites, therefore the work was initially focused on the process-phase relationship to identify the driving forces responsible to regulate the synthesis. Comparison of phase formation paths in conventional solid-state reaction and sol-gel method was conducted. The structural analyses revealed that intermediate pyrochlore K2Ta2O6 structure is not formed at any stage of the reaction using conventional solid-state reaction. On the other hand in the solution based processes, as alkoxide-based route, the crystallization of the perovskite occurs through the intermediate pyrochlore phase; at low temperatures pyrochlore is dominant and it is transformed to perovskite at >800 °C. The kinetic analysis carried out by using Johnson-MehlAvrami-Kolmogorow model and quantitative X-ray diffraction (XRD) demonstrated that in sol-gel derived powders the crystallization occurs in two stages: i) at early stage of the reaction dominated by primary nucleation, the mechanism is phase-boundary controlled, and ii) at the second stage the low value of Avrami exponent, n ~ 0.3, does not follow any reported category, thus not permitting an easy identification of the mechanism. Then, in collaboration with Prof. Alexandra Navrotsky group from the University of California at Davis (USA), thermodynamic studies were conducted, using high temperature oxide melt solution calorimetry. The enthalpies of formation of three structures: pyrochlore, perovskite and tetragonal tungsten bronze K6Ta10.8O30 (TTB) were calculated. The enthalpies of formation from corresponding oxides, ∆Hfox, for KTaO3, KTa2.2O6 and K6Ta10.8O30 are -203.63 ± 2.84 kJ/mol, - 358.02 ± 3.74 kJ/mol, and -1252.34 ± 10.10 kJ/mol, respectively, whereas from elements, ∆Hfel, for KTaO3, KTa2.2O6 and K6Ta10.8O30 are -1408.96 ± 3.73 kJ/mol, -2790.82 ± 6.06 kJ/mol, and -13393.04 ± 31.15 kJ/mol, respectively. The possible decomposition reactions of K-deficient KTa2.2O6 pyrochlore to KTaO3 perovskite and Ta2O5 (reaction 1) or to TTB K6Ta10.8O30 and Ta2O5 (reaction 2) were proposed, and the enthalpies were calculated to be 308.79 ± 4.41 kJ/mol and 895.79 ± 8.64 kJ/mol for reaction 1 and reaction 2, respectively. The reactions are strongly endothermic, indicating that these decompositions are energetically unfavourable, since it is unlikely that any entropy term could override such a large positive enthalpy. The energetic studies prove that pyrochlore is energetically more stable phase than perovskite at low temperature. Thus, the local order of the amorphous precipitates drives the crystallization into the most favourable structure that is the pyrochlore one with similar local organization; the distance between nearest neighbours in the amorphous or short-range ordered phase is very close to that in pyrochlore. Taking into account the stoichiometric deviation in KTO system, the selection of the most appropriate fabrication / deposition technique in thin films technology is a key issue, especially concerning complex ferroelectric oxides. Chemical solution deposition has been widely reported as a processing method to growth KTO thin films, but classical alkoxide route allows to crystallize perovskite phase at temperatures >800 °C, while the temperature endurance of platinized Si wafers is ~700 °C. Therefore, alternative diol-based routes, with distinct potassium carboxylate precursors, was developed aiming to stabilize the precursor solution, to avoid using toxic solvents and to decrease the crystallization temperature of the perovskite phase. Studies on powders revealed that in the case of KTOac (solution based on potassium acetate), a mixture of perovskite and pyrochlore phases is detected at temperature as low as 450 °C, and gradual transformation into monophasic perovskite structure occurs as temperature increases up to 750 °C, however the desired monophasic KTaO3 perovskite phase is not achieved. In the case of KTOacac (solution with potassium acetylacetonate), a broad peak is detected at temperatures <650 °C, characteristic of amorphous structures, while at higher temperatures diffraction lines from pyrochlore and perovskite phases are visible and a monophasic perovskite KTaO3 is formed at >700 °C. Infrared analysis indicated that the differences are due to a strong deformation of the carbonate-based structures upon heating. A series of thin films of alkali tantalates were spin-coated onto Si-based substrates using diol-based routes. Interestingly, monophasic perovskite KTaO3 films deposited using KTOacac solution were obtained at temperature as low as 650 °C; films were annealed in rapid thermal furnace in oxygen atmosphere for 5 min with heating rate 30 °C/sec. Other compositions of the tantalum based system as LiTaO3 (LTO) and NaTaO3 (NTO), were successfully derived as well, onto Si substrates at 650 °C as well. The ferroelectric character of LTO at room temperature was proved. Some of dielectric properties of KTO could not be measured in parallel capacitor configuration due to either substrate-film or filmelectrode interfaces. Thus, further studies have to be conducted to overcome this issue. Application-oriented studies have also been conducted; two case studies: i) photocatalytic activity of alkali tantalates and niobates for decomposition of pollutant, and ii) bioactivity of alkali tantalate ferroelectric films as functional coatings for bone regeneration. Much attention has been recently paid to develop new type of photocatalytic materials, and tantalum and niobium oxide based compositions have demonstrated to be active photocatalysts for water splitting due to high potential of the conduction bands. Thus, various powders of alkali tantalates and niobates families were tested as catalysts for methylene blue degradation. Results showed promising activities for some of the tested compounds, and KNbO3 is the most active among them, reaching over 50 % degradation of the dye after 7 h under UVA exposure. However further modifications of powders can improve the performance. In the context of bone regeneration, it is important to have platforms that with appropriate stimuli can support the attachment and direct the growth, proliferation and differentiation of the cells. In lieu of this here we exploited an alternative strategy for bone implants or repairs, based on charged mediating signals for bone regeneration. This strategy includes coating metallic 316L-type stainless steel (316L-SST) substrates with charged, functionalized via electrical charging or UV-light irradiation, ferroelectric LiTaO3 layers. It was demonstrated that the formation of surface calcium phosphates and protein adsorption is considerably enhanced for 316L-SST functionalized ferroelectric coatings. Our approach can be viewed as a set of guidelines for the development of platforms electrically functionalized that can stimulate tissue regeneration promoting direct integration of the implant in the host tissue by bone ingrowth and, hence contributing ultimately to reduce implant failure.

Cimentos injetáveis à base de fosfatos de cálcio para vertebroplastia

O principal objectivo desta investigação foi o desenvolvimento cimentos de fosfatos de cálcio com injetabilidade melhorada e propriedades mecânicas adequadas para aplicação em vertebroplastia. Os pós de fosfato de tricálcico (TCP) não dopados e dopados (Mg, Sr e Mn) usados neste estudo foram obtidos pelo processo de precipitação em meio aquoso, seguidos de tratamento térmico de forma a obter as fases pretendidas, α− e β−TCP. A substituição parcial de iões Ca por iões dopantes mostrou ter implicações em termos de estabilidade térmica da fase β−TCP. Os resultados demonstraram que as transformações de fase alotrópicas β↔α−TCP são fortemente influenciadas por variáveis experimentais como a taxa de arrefecimento, a presença de impurezas de pirofosfato de cálcio e a extensão do grau de dopagem com Mg. Os cimentos foram preparados através da mistura de pós, β−TCP (não dopados e dopados) e fosfato monocálcico monidratado (MCPM), com meios líquidos diferentes usando ácido cítrico e açucares (sucrose e frutose) como agentes retardadores de presa, e o polietilenoglicol, a hidroxipropilmetilcelulose e a polivinilpirrolidona como agentes gelificantes. Estes aditivos, principalmente o ácido cítrico, e o MCPM aumentam significativamente a força iónica do meio, influenciando a injetabilidade das pastas. Os resultados também mostraram que a distribuição de tamanho de partícula dos pós é um factor determinante na injetabilidade das pastas cimentícias. A combinação da co-dopagem de Mn e Sr com a adição de sucrose no líquido de presa e com uma distribuição de tamanho de partícula dos pós adequada resultou em cimentos de brushite com propriedades bastante melhoradas em termos de manuseamento, microestrutura, comportamento mecânico e biológico: (i) o tempo inicial de presa passou de ~3 min to ~9 min; (ii) as pastas cimentícias foram totalmente injectadas para uma razão liquido/pó de 0.28 mL g−1 com ausência do efeito de “filter-pressing” (separação de fases líquida e sólida); (iii) após imersão numa solução durante 48 h, as amostras de cimento molhadas apresentam uma porosidade total de ~32% e uma resistência a compressão de ~17 MPa, valor muito superior ao obtido para os cimentos sem açúcar não dopados (5 MPa) ou dopados só com Sr (10 MPa); e (iv) o desempenho biológico, incluindo a adesão e crescimento de células osteoblásticas na superfície do cimento, foi muito melhorado. Este conjunto de propriedades torna os cimentos excelentes para regeneração óssea e engenharia de tecidos, e muito promissores para aplicação em vertebroplastia.

Scaffolds porosos à base de fosfatos de cálcio para regeneração óssea

Graças ao aumento da esperança média de vida do ser humano, a engenharia de tecidos tem sido uma área alvo de enorme investigação. A utilização de estruturas tridimensionais porosas e biodegradáveis, denominadas de scaffolds, como matriz para a adesão e proliferação celular tem sido amplamente investigada. Existem atualmente diversas técnicas para a produção destas estruturas mas o grau de exigência tem vindo a aumentar, existindo ainda lacunas que necessitam ser preenchidas. A técnica de robocasting consiste numa deposição camada a camada de uma pasta coloidal, seguindo um modelo computorizado (CAD) e permite a produção de scaffolds com porosidade tamanho de poro e fração de porosidade controlados, boa reprodutibilidade, e com formas variadas, as quais podem ser idênticas às dos defeitos ósseos a preencher. O presente estudo teve como objetivo produzir scaffolds porosos à base de fosfatos de cálcio através de robocasting. Para tal, foram estudadas duas composições de pós à base de β-TCP, uma pura e outra co-dopada com estrôncio, zinco e manganês. Inicialmente os pós foram sintetizados pelo método de precipitação química por via húmida. Após a síntese, estes foram filtrados, secos, calcinados a 1000ºC e posteriormente moídos até possuírem um tamanho médio de partícula de cerca de 1,5 μm. Os pós foram depois peneirados com uma malha de 40μm e caracterizados. Posteriormente foram preparadas várias suspensões e avaliado o seu comportamento reológico, utilizando Targon 1128 como dispersante, Hidroxipropilmetilcelulose (HPMC) como ligante e polietilenimina (PEI) como agente floculante. Por fim, e escolhida a melhor composição para a formação da pasta, foram produzidos scaffolds com diferentes porosidades, num equipamento de deposição robótica (3D Inks, LLC). Os scaffolds obtidos foram secos à temperatura ambiente durante 48 horas, sinterizados a 1100ºC e posteriormente caracterizados por microscopia eletrónica de varrimento (SEM), avaliação dos tamanhos de poro, porosidade total e testes mecânicos. Ambas as composições estudadas puderam ser transformadas em pastas extrudíveis, mas a pasta da composição pura apresentou uma consistência mais próxima do ideal, tendo originado scaffolds de melhor qualidade em termos de microestrutura e de propriedades mecânicas.