O processo de cicatrização cutânea pode ser favorecido pela aplicação de laser de baixa potência, sendo a matriz de colágeno e elastina um substituto dérmico no tratamento de feridas descrita como um sistema adequado na engenharia tecidual em sistemas tridimensionais da pele. Com este intuito, foi elaborado um estudo utilizando o sistema nanoemulsão contendo cloro alumínio ftalocianina (ClAlPc) como o agente fotossensibilizante em biópsias de explants de pele de pacientes saudáveis irradiadas por luz proveniente de laser de baixa potência, visando estabelecer a melhor dose de luz para a bioestimulação de colágeno tipo I e elastina neste tecido. O sistema de liberação de fármacos foi sintetizado utilizando protocolos já conhecidos, cujas propriedades fotoquímicas e fotofísicas foram confirmadas por espectrofotometria de absorção e fluorescência, além de estudos de estabilidade medindo-se o tamanho das partículas, potencial Zeta e índice de polidispersão. Uma vez caracterizado este sistema e conhecendo-se a sua faixa de absorção, elaborou-se um protocolo para avaliar a ação do fármaco nos principais componentes da matriz extracelular, como colágeno e elastina, e a sua ação combinada com luz laser de baixa potência em três doses conhecidas distintas: 70, 140 e 700 mJ/cm2. Foi avaliada também a ação somente da irradiação sobre as biópsias, nas mesmas doses, podendo-se assim observar os seus efeitos. As estruturas morfológicas da pele foram estudadas por histologia, e posteriormente foram comparadas quantitativamente as porções de colágeno e elastina da pele tratada e irradiada com as amostras do controle, as quais não receberam nenhum tipo de tratamento. Tanto a análise para colágeno tipo I quanto a análise para elastina apontaram um aumento de quase 20% em relação às amostras não tratadas, utilizando a dose intermediária de 140 mJ/cm2 nas amostras tratadas com o fármaco de ftalocianina, durante um período de 14 dias após a irradiação. Este efeito foi bastante significativo, ao ser comparado com a ação somente da irradiação, que apresentou desempenho inferior. Outra técnica explorada neste trabalho e utilizada na detecção da expressão das enzimas MMP-2 e MMP-9, participantes do processo cicatricial, foi a zimografia de gelatina, utilizando o meio de cultivo de cada amostra. As bandas relacionadas à degradação da gelatina para cada amostra no zimograma foram quantificadas e os níveis de expressão de MMP-2 e MMP-9 comparados. Os resultados obtidos confirmaram a análise histológica, apontando um maior nível de expressão dessas enzimas nos grupos tratados com o fármaco fotossensibilizante e luz na dose intermediária (140 mJ/cm2) sendo esta combinação a mais promissora para um estudo mais aprofundado. / Wound healing process can be favored by Low Level Laser Therapy, with the collagen and elastin matrix a dermic substitute described as an appropriate system in tissue engineering and in tridimensional skin systems. With this purpose, a study was elaborated using the system nanoemulsion of aluminium-chloride phthalocyanine (ClAlPc) as photosensitizer in skin biopsies obtained after plastic surgery and irradiated by a low level laser, to establish the most appropriate dose of light for biostimulation of type I collagen and elastin fibers. The drug delivering system was synthesized using a well-known protocol and its photophysical and photochemical properties were confirmed by absorption and fluorescence spectrophotometry, besides stability studies measuring particle size, zeta potential and polidispersion index. Once characterized this system and known its absorption range in the UV-Vis region of light spectrum, a protocol was elaborated for evaluating the effect of the photosensitizer direct into extracellular matrix components, collagen and elastin, and the combined effect with low level laser therapy using three different doses: 70, 140 and 700 mJ/cm2. Also, only the effect of irradiation was evaluated using the same doses. Morphological structures of the skin were analyzed by histology, and portions of collagen and elastin of skin biopsies after the photosensitizer and light treatment were quantified and compared to the non-treated samples. The analysis for type I collagen and elastin pointed an increase of more than 20% compared to the non-treated samples, for the samples treated with the combination phosensitizer/light140 mJ/cm2 after 14 days of treatment. This effect was significant when compared to the effect of the irradiation only. Another technique was used in this work to detect the expression of MMP-2 and MMP-9, participant enzymes in various processes including tumoral progression and wound healing. Samples of biopsies culture medium were collected and analyzed by gelatin zymography, the bands related to gelatin degradation were quantified and MMP-2 and MMP-9 expression levels compared. The obtained results confirmed histological analysis, pointing a higher expression level of these enzymes for the group treated with the photosensitizer and the intermediate dose of light (140 mJ/cm2), leading to a promising combination of treatment for future studies.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Ectomycorrhizal fungi (fECM) combined with the plant root system improve the absorption of water and nutrients, providing great plant growth, especially in environments with nutritional deficiency or degraded. Due to the fECM and plants live in mutualistic association, allows the fungal symbionts to be benefit, or at least tolerate, the secondary metabolites produced by plants. It is possible that essential oils extracted from forest species that form mycorrhizal associations by stimulating the growth of ectomycorrhizal isolates and benefit the plant-fungus symbiosis. This study aimed to evaluate the effect of Eucalyptus grandis W. Hill ex. Maiden essential oil, the inoculum production of fECM and its influence on the tolerance of the isolates to heavy metals copper, zinc and nickel. Also, determines the effect of the use of essential oil on growth of eucalyptus (Eucalyptus grandis) and sibipiruna (Caesalpinia peltophoroides) seedlings in the greenhouse and the establishment of eucalyptus soil contaminated by copper under controlled conditions and field. The evaluation of the effect of different eucalyptus essential oil concentrations to stimulate the growth of fECM Pisolithus sp. (UFSC Pt 24 and UFSC Pt 188), Pisolithus microcarpus (UFSC Pt 116), Chondrogaster angustiporus (UFSC Ch 163), Scleroderma citrinum (UFSC Sc 124) and Suillus sp. (UFSM RA 2.2 and UFSM RA 2.8) in liquid culture medium MNM, allow to observe that the addition of eucalyptus essential oil concentrations 20 and 30 μL L-1 promoted an increase in mycelium growth in vitro of the isolates UFSC Pt 24, UFSC Pt 116, Ch 163 UFSC, UFSC Sc 124, UFSM RA 2.2 and UFSM RA 2.8. In liquid culture medium containing heavy metals copper, zinc and nickel. It was observed that the addition of essential oil at concentration of 20 μL L-1 increased the growth of ectomycorrhizal isolates. The evaluation of use of essential oil extracted from eucalyptus leaves on germination and initial development of eucalyptus seedlings in nursery conditions, permits to observe that germination was significantly highest when the seeds were sprayed with 25 μL L-1 oil essential. The application of essential oil at concentrations of 30 and 40 μL L-1 improve the development of roots and shoots of eucalyptus seedlings in a controlled environment. This can be an effective alternative in the biostimulation of the vegetative growth of eucalyptus seedlings. The effect of the use of essential oil was also positive when applied to eucalyptus seedlings inoculated 90 days after inoculation, improving both the mycorrhizal and seedling growth. Evaluating the effect of eucalyptus essential oil in the formation of ectomycorrhizal association and growth of sibipiruna seedlings after 90 days under greenhouse conditions. It was found that the application of essential oil enhanced the association of ectomycorrhizal fungus P. microcarpus and the development in seedlings of sibipiruna seedlings. The assessment of the degree of mycorrhizal association was determined by the percentage of mycorrhization. Eucalyptus seedlings grown in fertilized substrate, by using of essential oil in the formulation of inoculant or through subsequent application resulted in plants with highest height, fresh and dry weight, diameter and adaptability in soil contaminated by copper. These results may improve the establishment of mycorrhizal fungi in seedlings of sibipiruna seedlings as well as development and growth of eucalyptus seedlings in sites contaminated by heavy metals. / Os fungos ectomicorrízicos (fECM) em associação com o sistema radicular das plantas, auxiliam na absorção de água e nutrientes, proporcionando maior crescimento vegetal, principalmente em ambientes com deficiência nutricional ou degradados. Os fECM e as plantas ao conviverem em associação mutualística, permitem que os fungos simbiontes se beneficiem, ou pelo menos tolerem, os metabólitos secundários produzidos pelas plantas hospedeiras. É possível que os óleos essenciais extraídos de essências florestais conhecidamente micossimbiontes possam estimular o crescimento de isolados ectomicorrízicos e beneficiar a simbiose fungo-planta. O trabalho teve como objetivos determinar o efeito do óleo essencial de Eucalyptus grandis W. Hill ex. Maiden na produção de inóculo de fECM e sua influência na tolerância dos isolados aos metais pesados cobre, zinco e níquel; e determinar o efeito da utilização do óleo essencial no crescimento de mudas de eucalipto (Eucalyptus grandis) e sibipiruna (Caesalpinia peltophoroides) em casa de vegetação e no estabelecimento de eucalipto em solo contaminado por cobre em condições controladas e a campo. A avaliação do efeito de diferentes concentrações do óleo essencial de eucalipto no estímulo ao crescimento dos fECM Pisolithus sp. (isolados UFSC Pt 24 e UFSC Pt 188), Pisolithus microcarpus (isolado UFSC Pt 116), Chondrogaster angustiporus (isolado UFSC Ch 163), Scleroderma citrinum (isolado UFSC Sc 124) e Suillus sp. (isolados UFSM RA 2.2 e UFSM RA 2.8) em meio de cultura líquido MNM, permite observar que a adição do óleo essencial de E. grandis nas concentrações de 20 a 30 μL L-1 promoveu incremento no crescimento miceliano in vitro nos isolados UFSC Pt 24, UFSC Pt 116, UFSC Ch 163, UFSC Sc 124, UFSM RA 2.2 e UFSM RA 2.8. No meio de cultura líquido contendo os metais pesados cobre, zinco e níquel, observou-se que a adição do óleo essencial na concentração de 20 μL L-1 favoreceu o crescimento de isolados ectomicorrízicos. Avaliando-se a utilização do óleo essencial extraído de folhas de eucalipto sobre a germinação e o desenvolvimento inicial de mudas de eucalipto em condições de viveiro, observou-se que a germinação foi superior quando as sementes foram pulverizadas com 25 μL L-1 do óleo essencial. A aplicação do óleo essencial nas concentrações de 30 e 40 μL L-1, proporcionou maior desenvolvimento das raízes e da parte aérea das mudas de eucalipto em ambiente controlado. Isso pode ser uma alternativa eficiente na bioestimulação do crescimento vegetativo de mudas de eucalipto. O efeito da utilização do óleo essencial também foi positivo quando aplicado em mudas de eucalipto micorrizadas após 90 dias de inoculação, favorecendo tanto a micorrização como o crescimento das mudas. Avaliando-se o efeito do óleo essencial de eucalipto na formação de associação ectomicorrízica e no crescimento de mudas de sibipiruna após 90 dias em condições de casa de vegetação, verificou-se que a aplicação do óleo essencial favoreceu a associação ectomicorrízica, determinada através do porcentual de micorrização, do fungo P. microcarpus e o desenvolvimento em mudas de sibipiruna. Mudas de eucalipto formadas em substrato micorrizado, utilizando o óleo essencial na formulação do inoculante ou via aplicação posterior, resultaram em plantas com maior altura, massa fresca e seca, diâmetro do colo e capacidade de adaptação em solo contaminado por cobre. Esses resultados demonstram que a utilização do óleo essencial de eucalipto pode favorecer o estabelecimento de associação micorrízica em mudas de sibipiruna, bem como o crescimento e desenvolvimento de mudas de eucalipto em áreas contaminadas por metais pesados.

A thesis submitted to the College of Dental Medicine of Nova Southeastern University of the degree of Master of Science in Dentistry. Introduction: One of the challenges in orthodontics is obtaining desired tooth movement. A recent development in orthodontics offers patients more comfort by minimizing pain, and shortening the treatment time. Low level laser therapy (LLLT) emerged as a technology that may accelerate the velocity of tooth movement and shorten the orthodontic treatment period. LLLT (630-1000nm) has been shown to modulate various biological processes including wound healing and bone remodeling. Bone remodeling is one of the biological processes that ensue during tooth movement. Bone remodeling is a continuous process characterized by bone deposition at sites of tension and bone resorption on the pressure sites. At cellular level bone remodeling is regulated by receptor activator of NF-κB (RANK) and receptor activator of NF-κB ligand (RANKL) and Osteoprotegrin (OPG). RANK and RANKL promotes osteoclastic differentiation and promotes bone resorption. OPG is a soluble decoy receptor that competes with RANK for binding to RANKL and inhibits the osteoclastic activity. The goal of the research is to understand the regulatory effects of LLLT on bone metabolism at the cellular level. Furthermore, the purpose of this study is to evaluate the critical parameters of low level lasers on the early stage of ostegenic regulation of human osteoblast cells. In this study, CRL-1427 cells derived from human osteosarcoma which have an osteoblast phenotype were used as cell model. Methods: Human osteoblast cells CRL1427(ATCC, Manassas, VA) were cultured in minimum essential medium supplemented with 10% fetal bovine serum and 1% antibiotics and incubated in at 37°C with 5% CO2. The monolayer of cells after reaching 70-80% confluency were irradiated with a single dose of Galium Aluminum Arsenide (Ga-Al-As) laser, with a wave length of 830nm and output power of 85mW with 0.6, 1.5, 1.8 Joules/cm2 energy exposure regimes. The mRNA expressions of Alkaline phosphatase (ALP), Osteoprotegerin (OPG), RANKL and RANK were compared after seven days by quantitative real time PCR. Results: We observed that treatment of CRL 1427 cells with LLLT (1.5, 1.8 Joules/cm2) irradiation significantly increased the expression of ALP, OPG, RANKL and RANK mRNAs compared to the control group (P≤0.05). There was no significant difference between the control group and mRNA expression of ALP, OPG, RANK, or RANKL at 0.6J/cm2of irradiation. Conclusion: LLLT irradiation can directly influence the expression of genes associated with bone metabolism and potentially represents a mechanism that facilitates rapid bone remodeling.

This study was done to determine the effectiveness of a commercially available bioaugmentation product, BiOWiSHTM-Aqua FOG, for remediating petroleum-contaminated sandy soil. Biodegradation enhancement by BiOWiSHTM-Aqua FOG was evaluated in laboratory microcosms by directly measuring total petroleum hydrocarbon (TPH) and indirectly using respirometry. Attempts were made to enrich hydrocarbon-degrading bacteria in BiOWiSHTM-Aqua FOG, and the resulting enrichment cultures were screened using respirometry as well. Potential hydrocarbon-degrading bacteria in BiOWiSHTM-Aqua FOG were isolated. Experiments were performed at bench-scale using microcosm bottles containing sand contaminated with either motor oil or No. 2 diesel fuel. The microcosms were incubated at 25oC under aerobic conditions. TPH measurements of soil in the microcosms at 0, 25 and 56 days indicated that the addition of 500-ppm BiOWiSHTM-Aqua FOG improved biodegradation of the motor oil-contaminated soil by 45%. However, BiOWiSHTM-Aqua FOG did not have a measurable effect on biodegradation in the diesel-contaminated soil. In the respirometry experiments, BiOWiSHTM-Aqua FOG and two hydrocarbon-enriched BiOWiSHTM-Aqua FOG cultures were evaluated indirectly by the measurement of microbial carbon dioxide production and oxygen uptake using a MicroOxymaxTM respirometer. The respirometry experiments showed that in the six-day period following motor oil-contamination of soil, the addition of BiOWiSHTM-Aqua FOG substantially improves biodegradation rates. The added organisms in the product out-performed the indigenous organisms in the 5-6 days following contamination of the soil. The CO2 production observed in the BiOWiSHTM microcosms contaminated with motor oil was much greater than CO2 production without motor oil, which confirms that the observed metabolism can be attributed to motor oil biodegradation rather than metabolism of other organic material in the soil. Enriched consortia consistently generated far less CO2 than microcosms with the 500 ppm BiOWiSHTM-Aqua FOG. Stoichiometric calculations suggested that BiOWiSHTM-Aqua FOG removed approximately 1400 ppm TPH (14%) from the soil in 6.5 days, while an enrichment culture of BiOWiSHTM-Aqua FOG only reduced TPH levels by 459 ppm (5%). This result suggests that increased biodegradation rate in bioaugmented soil is aided by biodiversity in the augmenting inoculum. A potential hydrocarbon-degrading candidate organism was isolated from the product and cultured on Bushnell-Haas agar and plate-count agar (PCA). While at least two distinct colony types were successfully grown on media with motor oil, these same colonies appeared on Bushnell-Haas agar with no apparent carbon source, and survived repeated transfers onto this same medium. Therefore, their status as hydrocarbon-degraders is inconclusive. More thorough enrichment work could be pursued, especially using soil samples collected from petroleum-contaminated sites.

No description available.

In the last decades, nitrate pollution has become a major threat to groundwater quality. The consequences include health concerns and environmental impacts. Nitrate contamination is mainly derived from agricultural practices, such as the application of manure as fertilizer. The Osona area (NE Spain) is one of the areas vulnerable to nitrate pollution from agricultural sources. Nitrate is derived from intensive farming activities and the high nitrate content results in a loss of water availability for domestic uses. The most important natural nitrate attenuation process is denitrification. Denitrifying bacteria are generally heterotrophic and use carbon compounds as the electron donor. Nevertheless, a limited number of bacteria are able to carry out chemolithotrophic denitrification, and to utilize inorganic compounds. Several field studies have suggested by means of geochemical and/or isotopic data that denitrification in some aquifers is controlled by pyrite oxidation. However, the feasibility of pyrite-driven denitrification has been questioned several times in laboratory studies. This thesis is concerned with the role of pyrite in denitrification and its potential use as a bioremediation strategy. Earlier studies showed the occurrence of denitrification processes in a small area located in the northern part of the Osona region and suggested that sulfide oxidation had an important role in natural attenuation. Therefore, the first part of this thesis deals with the characterization of the denitrification processes occurring in the Osona aquifer and their spatial and temporal variations. Denitrification processes linked to pyrite oxidation were identified in some zones of the studied area by means of multi-isotopic methods integrated with classical hydrogeological methods. Nitrate removal from groundwater can be accomplished by the enhancement of in situ biological denitrification. One such bioremediation strategy is biostimulation, which involves the addition of suitable electron donors and/or energy sources to stimulate indigenous denitrifying microorganisms. The second part of this thesis is devoted to clarify the role of pyrite as electron donor for denitrification and to evaluate the feasibility of a bioremediation strategy based on pyrite addition to stimulate native denitrifying bacteria. Nitrate consumption in experiments amended with pyrite and inoculated with “Thiobacillus denitrificans” demonstrated that this bacterium is able to reduce nitrate using pyrite as the electron donor. The efficiency in nitrate removal and the nitrate reduction rate depended on the initial nitrate concentration, pH and pyrite grain size. High nitrate removal efficiency was attained in long-term flow-through experiments under laboratory conditions similar to those found in slow-moving, nitrate-contaminated groundwater. In addition, biostimulation experiments performed with sediments and groundwater from the Osona aquifer showed that the addition pyrite stimulated the activity of the indigenous microbial community and enhanced the nitrate removal. Furthermore, the long-term efficiency of the process was demonstrated. Hence, biostimulation with pyrite could be considered to remediate nitrate contamination in groundwater in future water management strategies, although further research is needed, especially at field scale. It is critical for the success of the bioremediation strategy that not only the processes but also the microbial populations and their changes induced by the bioremediation treatment be well understood. The addition of pyrite resulted in an increase in the proportion of denitrifying bacteria and both autotrophic and heterotrophic denitrifiers were stimulated. Bacterial populations closely related to the “Xanthomonadaceae” might probably be the dominant autotrophic denitrifiers that used pyrite as the electron donor in the biostimulated experiments. The N and O isotopic enrichment factors associated with the pyrite‐driven denitrification were computed and used to recalculate the extent of the natural nitrate attenuation in the Osona aquifer. This refinement becomes useful to predict the evolution of the contaminant in the aquifer, and it should be taken into account for potential implementation of induced remediation techniques. The isotopic approach was proved to be an excellent tool to identify and quantify natural denitrification processes in the field, and to monitor the efficacy of bioremediation strategies in the laboratory. In order to improve the long-term performance of potential bioremediation strategies based on pyrite-driven denitrification, it is necessary to know the contribution of attached and free-phase denitrifying bacteria to this process in aquifers. The last part of this thesis addresses the ability of “T. denitrificans” to grow and colonize pyrite surfaces. In the colonization experiments, attachment onto pyrite surface was required for at lest a small number of the cells in order to accomplish pyrite-driven denitrification. Nevertheless, both attached and planktonic cells probably contributed to the overall denitrification. However, the details of the relative roles of the two phases and the specific mechanisms remain to be addressed. / A les darreres dècades, la contaminació de nitrat de l’aigua subterrània ha esdevingut un dels principals problemes que afecten la qualitat dels recursos hídrics subterranis. La presència de nitrat a l’aigua subterrània està relacionada, principalment, amb pràctiques agrícoles, com per exemple, l’ús intensiu de purins com a fertilitzant orgànic. La comarca d’Osona (Catalunya) és una de les àrees declarades vulnerables a la contaminació per nitrat d’origen agrícola. El nitrat procedeix principalment de la intensa activitat agrícola i ramadera. El principal procés d’atenuació natural del nitrat és la desnitrificació. La majoria dels bacteris desnitrificants són heteròtrofs i usen compostos orgànics com a font d’electrons. Tanmateix, també existeixen bacteris desnitrificants autòtrofs que són capaços d’usar compostos inorgànics, com a donants d’electrons. Estudis de camp previs han demostrat a partir de dades geoquímiques i/o isotòpiques que en alguns aqüífers la desnitrificació està controlada per l’oxidació de pirita. Tot i així, hi ha estudis de laboratori que qüestionen la viabilitat de la desnitrificació lligada a l’oxidació de pirita (compost inorgànic donant d’electrons). En aquesta tesi s’estudia el paper de la pirita en la desnitrificació i la seva possible aplicació com a estratègia de bioremediació d’aigües contaminades amb nitrat. Estudis previs demostraren l’ocurrència de processos de desnitrificació a Osona, en una petita àrea situada en el sector nord de la zona d’estudi. Els resultats d’aquests estudis suggereixen que l’oxidació de sulfurs hi juga un paper molt important en l’atenuació natural. A la primera part de la tesi s’exposa la caracterització dels processos de desnitrificació que tenen lloc en l’aqüífer d’Osona, així com de les seves variacions temporals i espacials. S’han usat mètodes multi-isotòpics integrats amb mètodes hidrogeològics clàssics, que han servit per identificar l’existència de processos de desnitrificació lligats a l’oxidació de pirita en algunes zones de l’àrea d’estudi. L’eliminació del nitrat de l’aigua subterrània pot aconseguir-se mitjançant estratègies que incentiven la desnitrificació biològica in situ. Una de les estratègies de bioremediació in situ és la bioestimulació, que consisteix en afegir donants d’electrons o fonts d’energia apropiades per tal d’estimular l’activitat de bacteris desnitrificants existents en el medi. En la segona part de la tesi s’esbrina el paper de la pirita com a potencial donant d’electrons en els procés de desnitrificació i s’avalua la viabilitat d’una estratègia de bioremediació, basada en l’addició de pirita per estimular l’activitat de bacteris desnitrificants autòctons. Experiments amb pirita i inoculats amb “Thiobacillus denitrificans” han demostrat que aquest bacteri és capaç de reduir el nitrat utilitzant la pirita com a donant d’electrons. A més a més, s’ha determinat que l’eficiència i la velocitat de reducció del nitrat depèn de la seva concentració inicial, del pH i de la mida de gra de la pirita. En experiments de flux continu, de llarga durada que simulen el flux d’aigües subterrànies contaminades amb nitrat, s’han aconseguit altes eficiències en l’eliminació del nitrat. A més a més, s’han realitzat experiments de bioestimulación amb pirita usant aigua subterrània i sediments de l’aqüífer d’Osona. Aquests experiments han demostrat que afegint pirita s’aconsegueix d’estimular l’activitat dels bacteris desnitrificants autòctons i, per tant, induir i/o augmentar la desnitrificació. També, s’ha provat l’alta eficiència d’aquest procés a llarg termini. Per tant, l’ús de la pirita podria tenir-se en compte com a estratègia per a l’eliminació de nitrat en properes mesures de gestió dels recursos hídrics. Tanmateix, cal més investigació, i especialment, a escala de camp. Per aconseguir l’èxit amb la bioremediació cal conèixer amb detall els processos i les poblacions microbiològiques que intervenen en els canvis poblacionals associats al tractament de bioremediació. Els resultats dels experiments de bioestimulació han demostrat que els bacteris desnitrificants autòtrofs dominants en el sistema són probablement poblacions relacionades amb Xanthomonadaceae. L’addició de pirita puposa un increment en la proporció de bacteris desnitrificants i produeix l’estimulació tant dels desnitrificants autòtrofs com dels heteròtrofs. A més a més, s’han calculat els factors d’enriquiment isotòpic de N i O associats al procés de desnitrificació per oxidació de pirita. Aquests factors d’enriquiment s’han usat per recalcular el grau d’atenuació natural del nitrat que té lloc en l’aqüífer d’Osona. Aquest recàlcul pot ser d’utilitat a l’hora de predir l’evolució de la contaminació en l’aqüífer i cal tenir-lo en compte a l’hora d’implementar possibles tècniques de remediació induïda. Per tant, la metodologia isotòpica ha demostrat ser una eina excel∙lent per identificar i quantificar processos de desnitrificació natural i per monitoritzar l’eficàcia d’estratègies de bioremediació en el laboratori. Per millorar el rendiment i la durabilitat de potencials estratègies de bioremediació basades en la desnitrificació i oxidació de pirita és necessari conèixer la contribució relativa dels bacteris desnitrificants planctònics i dels bacteris adherits als sediments. Així, a l’última part de la tesi l’interès s’enfoca en l’avaluació de l’habilitat de “T. denitrificans” en colonitzar i créixer sobre la superfície de la pirita. Els resultats experimentals de colonització suggereixen que per aconseguir la desnitrificació amb pirita com a donant d’electrons, cal que una part dels bacteris s’adhereixi a la superfície de la pirita. No obstant, sembla que ambdós bacteris, adherits i planctònics, contribueixen al procés global. Tanmateix, queda pendent d’examinar el paper específic de cada tipus en la desnitrificació.

Orientador: Rosana Rossi Ferreira / Resumo: A medicina regenerativa é uma área em crescente expansão no Brasil e no mundo, a qual procura ampliar a capacidade natural de regeneração dos tecidos através da utilização de células, fatores de proliferação e biomateriais. Um dos ramos da medicina regenerativa é a terapia celular, vertente que utiliza células-tronco, visando a substituição de tecidos funcionalmente ou estruturalmente lesados, apresentando um caráter terapêutico. Na medicina LASERs e LEDs vem sendo estudados como ferramenta terapêutica, mostrando possuir capacidade bioestimulatória. Este campo é caracterizado por uma variedade de metodologias, que são utilizadas em uma gama considerável de aplicações. Na técnica de fotoestimulação, utiliza-se a luz para ativar moléculas e funções celulares, apresentando o potencial de afetar a proliferação e diferenciação e o metabolismo da célula, estimulando a fosforilação oxidativa e podendo reduzir a resposta inflamatória local. Entretanto para que essa resposta ocorra, inúmeros trabalhos afirmam sobre a importância da seleção de um comprimento de onda ideal, uma vez que a utilização de um comprimento inapropriado pode acarretar em resultados contrários aos esperados, como a bioinibição. Diante destes achados o presente trabalho propôs-se a avaliar a ação do LED 850nm, no regime pulsátil, nas doses de 3, 5 e 10J/cm² na cultura de célula-tronco mesenquimal (CTM) com Soro Fetal Bovino (SFB) e com Hormônios derivados de plaquetas (HDP), e na cultura de células de Linfoma lin... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Regenerative medicine is a promising growing area worldwide, with the aim of restore and regenerate tissues and whole organs through the use of cells, proliferation factors and biomaterials. One branch of regenerative medicine is cell therapy, that uses stem cells, aiming at the substitution of functionally or structurally damaged tissues, presenting therapeutic fature. LASERs and LEDs are available as therapeutic tools, showing biostimulating ability. The photo-stimulation technique uses light to activate molecules and cellular functions, presenting potential to affect proliferation, cell differentiation and metabolism, stimulating oxidative phosphorylation and reducing the local inflammatory response. Data shows the importance of selecting an ideal wavelength, such as the use of an inappropriate choice, can lead to undisered results, such as bioinhibition. In the present work, we evaluated the action of LED 850nm, pulsatile, at the doses of 3, 5 and 10J/cm² in mesenchymal stem cells (CTM) with Bovine Fetal Serum (FBS) and with derived platelets – Hormones (HDP) and B-cell lymphoblastic cell culture, 10J /cm2 , RAJI cells. In all light exposure experiments, wavelength of 850 nm inhibited cell proliferation. CTM culture, LED had a low proliferation rate, resulting in a decrease in cellular confluence, especially at 5 and 10J/cm2 . Lymphoblastic lymphoma type B cells, in only one week of exposure presente the same behavior of bioinhibition at 10J/cm2 . The control group had 7.... (Complete abstract click electronic access below) / Mestre

Submitted by bruna ortiz (brunaortiz.f@gmail.com) on 2016-07-04T14:19:54Z No. of bitstreams: 1 Dissertação- Siomara Dias da Rocha.pdf: 4062396 bytes, checksum: 46a7bfa943dd6bdb2560140dd4db722f (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-07-21T13:08:59Z (GMT) No. of bitstreams: 1 Dissertação- Siomara Dias da Rocha.pdf: 4062396 bytes, checksum: 46a7bfa943dd6bdb2560140dd4db722f (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-07-21T13:11:32Z (GMT) No. of bitstreams: 1 Dissertação- Siomara Dias da Rocha.pdf: 4062396 bytes, checksum: 46a7bfa943dd6bdb2560140dd4db722f (MD5) / Made available in DSpace on 2016-07-21T13:11:32Z (GMT). No. of bitstreams: 1 Dissertação- Siomara Dias da Rocha.pdf: 4062396 bytes, checksum: 46a7bfa943dd6bdb2560140dd4db722f (MD5) Previous issue date: 2015-12-18 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / The interest in the study of environmental contamination by polycyclic aromatic hydrocarbons (PAH) lies in the fact that most of these compounds are potentially carcinogenic and mutagenic. Microorganisms play an important role, since they use these substances as a source of energy. In this study, the fungal isolates of the macrophyte Eichhornia crassipes (Mart) Solms were selected in medium BH (Bushnell Haas) containing the crude technique based on redox indicator 2, 6 - dichlorophenol indophenol (DCPIP), by means of oxidation occurring biological indicated by a change in coloring of the culture medium. Standing out this test, the fungi encoded F13, F14 and F01, respectively, Phoma herbarum, Fusarium moniliforme and Penicillium citrinum, which exhibited biological activity between 24 and 48 hours. In order to contribute to the advancement in research on bioremediation this work developed studies on hydrocarbon degradation using fungal consortium in liquid mineral medium (BH) supplemented with 15 uL of crude oil using bioaugmentation and biostimulation tests in different biodegradation times: time 0, 48 hours, 96 hours and 7 days. To evaluate the oil degradation behavior by fungi were performed by quantitative analysis of polycyclic aromatic hydrocarbons (PAH) from the application of analytical methods: liquid-liquid extraction of the sample supernatant in the presence of oil (in accordance with EPA Method 3510C) and under the action of fungal consortium. Then there was the cleaning step and fractionation of the sample in open column (EPA Method 8270D modified) obtaining a fraction of the HPA for the analysis of 16 compounds, considered of medium polarity and established as priorities by the Environmental Protection Agency States (US EPA); and as a last step: separation and quantification of hydrocarbons by gas chromatography-mass spectrometry (GC-MS). The analytical curves showed determination coefficient values (R2) above 0.99 for the 16 HPA. The mean recovery values for p-terphenyl surrogate added to the samples before the extraction stage were in the range of 67.15/99.06%. The investigation of the possible degradation of these recalcitrant compounds demonstrate the potential of these fungi in a consortium, which was observed during the 7 days of the experiment and the decrease in individual concentration of the analyzed compounds. These results allow to observe the crude oil degradation behavior and infer that the samples with pH correction acid without additional source of nutrients showed to be more efficient in the degradation of HPA 16 within 7 days with the reduced concentration of between 179.56 ± 0.07 to 77,00 ± 0.21 ng mL-1 equivalent to 64% degradation. The normality test of Shapiro-Wilk proved that the data does not follow a normal distribution, with p-value 0.02 (<0.05). The ANOVA analysis showed that time directly influences the degradation. It is believed that this study contributes to the numerous studies on bioremediation in the Amazon in order to advance the research on the reduction of environmental contaminants, particularly aromatic hydrocarbons. / O interesse pelo estudo da contaminação ambiental por hidrocarbonetos policíclicos aromáticos (HPA) reside no fato de que a maioria destes compostos são potencialmente carcinogênicos e mutagênicos. Os micro-organismos desempenham um importante papel, uma vez que utilizam essas substâncias como fonte de energia. No presente estudo, os fungos isolados da macrófita Eichhornia crassipes (Mart.) Solms foram selecionados em meio BH (Bushnell Haas) contendo petróleo bruto pela técnica baseada no indicador redox 2,6 - diclorofenol indofenol (DCPIP), por meio da ocorrência de oxidação biológica indicada pela mudança na coloração do meio de cultivo. Destacando-se neste teste, os fungos codificados F13, F14 e F01, respectivamente: Phoma herbarum, Fusarium moniliforme e Penicillium citrinum, que apresentaram atividade biológica entre 24 e 48 horas. Com o intuito de contribuir para o avanço nas pesquisas sobre biorremediação este trabalho desenvolveu estudos sobre degradação de hidrocarbonetos utilizando consórcio fúngico em meio mineral líquido (BH) suplementado com 15 μL de petróleo bruto utilizando ensaios de bioaumento e bioestímulo em diferentes tempos de biodegradação: tempo 0, 48 horas, 96 horas e 7 dias. Para avaliar o comportamento da degradação do petróleo pelos fungos foram realizadas análises quantitativas dos hidrocarbonetos policíclicos aromáticos (HPA) a partir da aplicação de métodos analíticos: extração líquido-líquido do sobrenadante da amostra na presença de petróleo (prevista no Método EPA 3510C) e sob a ação do consórcio fúngico; em seguida realizou-se a etapa de limpeza e fracionamento da amostra em coluna aberta (Método EPA 8270D modificado) obtendo-se a fração dos HPA para análise dos 16 compostos, considerados de média polaridade e estabelecidos como prioritários pela Agência de Proteção Ambiental dos Estados Unidos (US EPA); e como última etapa: separação e quantificação dos hidrocarbonetos por cromatografia gasosa acoplada à espectrometria de massas (CG-EM). As curvas analíticas apresentaram valores de coeficiente de determinação (R2) acima de 0,99 para os 16 HPA. Os valores médios de recuperação para o surrogate p-terfenil adicionado nas amostras antes da etapa de extração ficaram na faixa de 67,15 a 99,06%. A investigação da biodegradação desses compostos recalcitrantes possibilitou evidenciar o potencial desses fungos em consórcio, fato observado no decorrer dos 7 dias do experimento e pela diminuição da concentração individual dos compostos analisados. Esses resultados possibilitaram observar o comportamento da degradação do óleo bruto e inferir que as amostras com correção de pH ácido sem fonte adicional de nutrientes apresentou-se mais eficiente na degradação dos 16 HPA em até 7 dias com a redução da concentração variando entre 179,56 ± 0,07 a 77,00 ± 0,21 ng.mL-1 equivalente a 64% de degradação. O teste de normalidade de Shapiro-Wilk comprovou que os dados não seguem uma distribuição normal, com p-valor 0,02 (<0,05). A análise ANOVA mostrou que o tempo influencia diretamente na degradação. Acredita-se que este estudo contribua com os inúmeros trabalhos sobre biorremediação na Amazônia com finalidade de avançar nas pesquisas sobre a redução de contaminantes ambientais, em especial hidrocarbonetos aromáticos.

O objetivo do trabalho foi identificar ferramentas e indicadores adequados ao monitoramento e à otimização de processos de biorremediação, incluindo parâmetros físicos, químicos e microbiológicos definidos em estudos de tratabilidade de solos contaminados por óleo cru em escala de laboratório e comparar estratégias de biorremediação, tais como bioestímulo e bioaumento conduzidas em simulações de biopilhas dinâmicas ou estáticas. Quando três métodos de extração de hidrocarbonetos de petróleo de solo arenoso e franco-argiloso para análise cromatográfica (Soxhlet-SOX, microondas-MARS e extração acelerada por solvente-ASE) foram comparados entre si, concluiu-se que a técnica que promove a melhor recuperação depende da fração de interesse (n-alcanos, HRP, MCNR, HPA), das características texturais do solo (teores de areia, silte e argila) e da idade da contaminação. Dentre os indicadores de densidade populacional microbiana (microrganismos heterotróficos totais-PHT, população de fungos-PF e população microbiana degradadora de óleo (PDO) passíveis de utilização para indicar a taxa de degradação de compostos orgânicos presentes no solo tais como os hidrocarbonetos de petróleo, o PDO mostrou-se o mais adequado em conjunto com a produção de CO2 aferida pelo método respirométrico. Quando a estratégia de biorremediação de solo franco-argiloso contaminado com óleo cru a 3% (m m-1) utilizando bioestímulo (ajuste de pH, umidade e taxa C:N:P) foi comparada ao bioaumento (bioestímulo e adição de inóculo de microrganismos extraídos, enriquecidos e aclimatizados ao óleo cru como fonte de carbono), em sistemas de bancada simulando biopilha dinâmica (microcosmo M) e biopilha estática com aeração forçada (reator B), o tratamento que apresentou melhor remoção (32%) de HTP após 121 dias foi o bioaumento em biopilha estática. Para HPA, o tratamento que alcançou a melhor remoção (33%) foi com bioestímulo também em biopilha estática. A avaliação da taxa de mortalidade (%) de Eisenia andrei exposta tanto a solos recém-contaminados por óleo cru e preparados para bioestímulo (BIOS) e bioaumento (BIOA) a serem tratados em biopilhas dinâmicas e estáticas em escala de laboratório mostrou que após 56 dias de exposição da E. andrei, todos os solos produziram letalidade de 100%, quer fossem os solos recém-contaminados e preparados para os diferentes tratamentos (BIOS M, BIOS B, BIOA M, BIOA B) ou após 121 dias de tratamento. Tal resultado confirma que a biorremediação foi incipiente também do ponto de vista de remoção da ecotoxicidade. Em linhas gerais, a biorremediação de solo franco-argiloso contaminado por óleo cru, contendo tanto contaminação antiga quanto recente, reúne os maiores desafios à biorremediação, tanto do ponto de vista da composição textural do solo quanto da natureza do contaminante. Os processos são aparentemente lentos e requerem ferramentas auxiliares para aceleração dos mesmos. Recomenda-se no futuro, condução de experimentos com o uso de diferentes surfactantes, com ênfase em biosurfactantes / The objective of this investigation was to identify tools and indicators suitable for monitoring and optimizing bioremediation processes, including physical, chemical and microbiological variables defined in treatability studies of crude oil contaminated clayey soils in lab-scale and to compare bioremediation strategies, such as biostimulation and bioaugmentation carried out in lab scale dynamics and static biopiles. When three methods for extraction of petroleum hydrocarbon for chromatography analysis (Soxhlet-SOX, microwaves-MARS and accelerated solvent extraction-ASE) were compared, it was concluded that the most appropriate extraction method depends on the hydrocarbon fraction of interest (n-alkanes, RPH, UCM or PAH), the textural soil characteristics (sand, silt and clay content) and the contamination age. Among the microbiological indicators investigated (total heterotrophic microorganisms-PHT, fungi population-PF and; oil-degrading microorganism population-PDO, the last one was the best indicator for degradation of organic carbon compounds such as petroleum hydrocarbon, together with CO2 generation measured through respirometric method. Bioremediation strategies for oil-contaminated clayey soil (3% m m-1) were compared, including biostimulation (adjustment of pH, C:N:P rate and moisture) and bioaugmentation (biostimulation plus a consortium inoculum extracted from the original aged-contaminated soil, enriched and acclimatized with the crude oil as carbon source) in lab scale system simulating dynamic (microcosms) and static (bioreactors with forced aeration). The treatment which presented the highest TPH removal (32%) after 121 days was the bioaugmented static biopile (BIOA B). For PAH, the most efficient treatment method that removed 33% of PAH was the biostimulated static biopile (BIOS B). The mortality rate (%) for Eisenia andrei exposed to newly contaminated soil prepared for biostimulation (BIOS) and bioaugmentation (BIOA) treatments at day 0 as well as the treated soil in static and dynamic biopiles at different treatment times (after 24, 59 and 121 days) revealed that after 56 of E. andreis exposition, all soils produced mortality of 100%, no matter if newly contaminated or treated (BIOS M, BIOS B, BIOA M, BIOA B). The ecotoxicity tests confirmed that bioremediation as such was incipient also from the toxicity removal viewpoint. In general terms, bioremediation of clayey soils with a complex contaminant such as crude oil raise considerable challenges, both from the soil characteristics as well as the nature of the contaminant. The processes are apparently slow and require auxiliary tools to speed up the process. In the future, it is recommended to test the use of different surfactants, particularly bio-surfactants.

Submitted by Marcio Emanuel Paixão Santos (marcio.santos@ucsal.br) on 2017-02-18T14:18:59Z No. of bitstreams: 1 DISSERTACAO FINAL_GUIOMAR SANTIAGO_ 2012.pdf: 3848547 bytes, checksum: dc81b0dad3448700a07e9d0e1beff845 (MD5) / Rejected by Maria Emília Carvalho Ribeiro (maria.ribeiro@ucsal.br), reason: Correções on 2017-02-22T18:22:51Z (GMT) / Submitted by Marcio Emanuel Paixão Santos (marcio.santos@ucsal.br) on 2017-02-22T22:37:49Z No. of bitstreams: 1 DISSERTACAO FINAL_GUIOMAR SANTIAGO_ 2012.pdf: 3848547 bytes, checksum: dc81b0dad3448700a07e9d0e1beff845 (MD5) / Approved for entry into archive by Maria Emília Carvalho Ribeiro (maria.ribeiro@ucsal.br) on 2017-03-02T21:14:30Z (GMT) No. of bitstreams: 1 DISSERTACAO FINAL_GUIOMAR SANTIAGO_ 2012.pdf: 3848547 bytes, checksum: dc81b0dad3448700a07e9d0e1beff845 (MD5) / Made available in DSpace on 2017-03-02T21:14:30Z (GMT). No. of bitstreams: 1 DISSERTACAO FINAL_GUIOMAR SANTIAGO_ 2012.pdf: 3848547 bytes, checksum: dc81b0dad3448700a07e9d0e1beff845 (MD5) Previous issue date: 2012-09 / Este trabalho analisou as estruturas existentes na Baía de Todos os Santos para atendimento à emergências ambientais por vazamento de petróleo, analisou a aplicação das técnicas de biorremediação de manguezais impactados por hidrocarbonetos de petróleo, caso atingidos por vazamento, assim como a participação da comunidade em processos de recuperação dos manguezais e para isso analisou a percepção da comunidade de marisqueiros da comunidade de Mataripe, distrito de São Francisco do Conde - BA, quanto aos impactos existentes sobre os manguezais da região estudada. A Baía de Todos os Santos tem seu Plano de Área (plano de contingência com as estratégias de resposta a emergências ambientais) em elaboração. A análise das técnicas de biorremediação mostrou a bioestimulação e a fitorremediação como as técnicas que melhor otimiza o binômio custo-benefício. A pesquisa realizada indica que a comunidade de Mataripe – São Francisco do Conde apesar da proximidade às áreas de produção, refino e portuária existentes na região nordeste da Baía de Todos os Santos, percebe a própria comunidade como um dos fatores principais de degradação dos manguezais e mostra-se solícita a participar de atividades que melhorem a realidade e condições dos manguezais locais, como atividades de educação ambiental e de recuperação de manguezais, o que é viável e pode ser uma nova fonte de renda para as pessoas da comunidade. / This paper analyzed the existing structures in the All Saints Bay for compliance at oil spill environmental emergencies, considered the application of bioremediation techniques of mangroves impacted by petroleum hydrocarbons, as well as community participation in mangroves recovery. It was examined the perception of shellfish community in the Mataripe, district of São Francisco do Conde - BA, related as mangroves impacts of that region. Now a day the Area Plan (contingency plan with strategies for responding to environmental emergencies) is been elaborated. The analysis of bioremediation techniques showed biostimulation and phytoremediation as techniques that best optimizes the cost-benefit ratio. The research indicates that community Mataripe - São Francisco do Conde, despite the proximity to the areas of production, and refining existing port in the northeast of Todos os Santos Bay, is the mainly factor at the mangroves degradation however, they are ready to participate in activities that turn best the local conditions and the mangroves, for example environmental education and restoration of mangroves activities, something feasible that can be a new source of income for people in the community.