Introdução sistemas de tratamento de esgotos, quando aplicados ao tratamento de lixiviado de aterro sanitário, não têm apresentado bons resultados. Por conta disso, faz-se necessário o desenvolvimento de alternativas para o tratamento de lixiviado. Objetivo avaliar o pós-tratamento de lixiviado de aterro sanitário por wetland construído de fluxo subsuperficial horizontal e reator biológico de leito móvel (RBLM). Materiais e métodos foram realizados dois experimentos com tratamento de lixiviado após remoção de amônia por dessorção. No experimento 1 utilizou-se wetlands construídos de fluxo horizontal em escala de bancada (volume total de 30,8 L), plantados com Cyperus papyrus, Heliconia psittacorum e Gynerium sagittatum, e um controle sem vegetação, preenchidos com pedrisco calcário. O sistema foi alimentado com lixiviado diluído em água em diferentes proporções (entre 10 por cento e 30 por cento ) com concentração média de DQO entre 336 e 750 mg.L-1 e nitrogênio amoniacal (N-NH4) entre 47 e 199 mg.L-1. A operação ocorreu em três etapas: (1ª) alimentação contínua e TDH médio entre 2,7 e 5,3 d; (2ª) regime de ciclos de recirculação do efluente, para avaliar o efeito do aumento de TDH para 21 dias; (3ª) alimentação contínua de duas unidades wetlands em série, visando elevar o TDH (entre 8,1 e 9,9 d) sem recirculação. O experimento 2 foi implantado em escala piloto, com uma unidade de dessorção de amônia seguida de um Reator Biológico de Leito Móvel (RBLM) e um biofiltro anóxico. O RBLM (volume efetivo de 380 L) foi preenchido em 50 por cento com anel pall. Resultados e discussão No experimento 1, obteve-se baixa remoção de DQO com médias inferiores a 40 por cento e concentração final entre 270 e 750 mg.L-1. No entanto, verificou-se remoção de N-NH4, principalmente na 2ª e 3ª etapas, com médias de remoção entre 43 por cento e 81 por cento , resultando em concentração de 20 a 223 mg.L-1, com influência do TDH. No experimento 2, problemas técnicos durante a instalação e operação inicial e interdição do campus da USP-Leste prejudicaram a adaptação do reator e impediram o seu monitoramento por tempo suficiente para geração de dados consistentes, sendo interrompido na fase inicial. Conclusões A baixa eficiência para DQO provavelmente está relacionada à recalcitrância do lixiviado, ou devido à inibição dos microrganismos por toxicidade. Quanto ao N-NH4, infere-se que o aumento do TDH proporcionou maior tempo de difusão de oxigênio para o sistema wetland, suprindo a demanda das bactérias heterotróficas, o que possibilitou utilização do oxigênio excedente pelas nitrificantes, culminando na oxidação do N-NH4. / Introduction sewage treatment systems, when applied to the treatment of landfill leachate, have not shown good results. Thus it is necessary to develop alternatives for the treatment of leachate. Objective To assess the post-treatment of landfill leachate by horizontal subsurface flow constructed wetland and moving bed biofilm reactor (MBBR). Materials and methods Two experiments were conducted with treatment of the leachate after removal of ammonia by stripping. In experiment 1, horizontal flow constructed wetlands, at bench scale (total volume of 30.8 L), planted with Cyperus papyrus, Heliconia psittacorum and Gynerium sagittatum, and a control without vegetation, were filled with calcareous gravel. The system was fed with leachate diluted in water with different proportions (between 10 per cent and 30 per cent ) with mean COD concentration of between 336 and 750 mg.L-1 and ammonia nitrogen (NH4-N) between 47 and 199 mg L-1. The operation was divided in three stages: (1st) continuous feeding with mean HRT between 2.7 and 5.3 d; (2nd) regime of effluent recirculation to evaluate the effect of increasing the HRT to 21 days; (3rd) continuous feeding with two wetlands units in series, aimed HRT increasing (between 8.1 and 9.9 d) without recirculation. Experiment 2 was implemented on a pilot scale, with a unit of ammonia stripping followed by MBBR and an anoxic biofilter. The MBBR (380 L working volume) was filled 50 per cent with pall ring. Results and discussion In experiment 1, the COD removal was low, with averages below 40 per cent , with effluent concentration between 270 and 750 mg.L-1. However, there was removal of NH4-N, especially in the 2nd and 3rd stages, with average removal between 43 per cent and 81 per cent , resulting in concentration between 20 and 223 mg.L-1, with HRT influence. In experiment 2, technical problems during the installation and initial operation and interdiction of the campus of USP-Leste disrupted the reactor acclimation and precluded the monitoring for sufficient time to generate consistent data for discussion. Conclusions The low efficiency for COD is probably related to the recalcitrance of the leachate, or due to inhibition of microorganisms by toxicity. For NH4-N, it is inferred that the increase of the HRT provided greater time for diffusion of oxygen to the wetland system, meeting the demand of heterotrophic bacteria, which allowed the use of excess oxygen by nitrifiers, culminating in the oxidation of NH4-N.

Trä är den bästa förnyelsebara källan för att producera många produkter på grund av dess biokompatibla och biologiskt nedbrytbara natur. Träets biomassa har en motstridig natur mot enzymatisk uppgradering. Det beror på olika orsaker som ligninhalt, acetylhalt i hemicellulosa och cellulosakristallinitet som blockerar enzymets bindningsställe. Denna studie kommer under BioUPGRADE, som är en samarbetsplattform för att skapa högvärdiga och mångsidiga material på ett hållbart sätt med hjälp av biokatalys. Det allmänna syftet med denna studie är att producera holocellulosa och hemicellulosasubstrat från olika träslag och producera, rena och validera galaktosoxidas som en potentiell biokatalysator för träfibermodifiering. Studien undersöker effekten av kemisk perättiksyradelignifiering på två träslag, lövträ av eukalyptus (HW), och barrträ av gran (SW), undersökta vid olika tidsintervall, där PAA framställdes exsitu med ett volymetriskt förhållande på 1:3. Med resultaten från PAA-behandlingen avlägsnades 38,53 % lignin i eukalyptus och 31,80 % i barrved. Hemicellulosautbytet ökade med 47,40 % för eukalyptus och 19,05 % för gran med en ökning av tiden för PAA-behandling. Acetylhalten i hemicellulosan minskade från 2 % till 0,6 % i lövträ och 1,96 % till 0,6 % i barrträ. Cellulosautvinningen efter delignifieringen var nästan 100 %. Galaktosoxidaset producerades i en skakkolv med användning av Pichia pastoris KM71H-stammen. Pichia pastoris KM71H-celler odlades med användning av det buffrade komplexa glycerolmediet (BMGY) och galaktosoxidas uttrycktes med användning av Pichia pastoris KM71H-stammen i buffrat komplex metanolmedium (BMMY). Det uttryckta GaOx-proteinet renades därefter med användning av AKTA-kromatografi med användning av en 5 ml Histrap FF-kolonn. För att bestämma proteinkoncentrationen utfördes bicinchoninsyra (BCA) analys och GaOx som producerades i skakkolvsodling var 286,25 mg/L. Den specifika aktiviteten hos skakkolven som produceras GaOx är 164,24 U/mg. Det kan observeras att PAA-behandling visar sig vara en effektiv metod för delignifiering eftersom cellulosautvinningen är nära 100 % och förlusten av hemicellulosa är relativt låg med det använda volymetriska förhållandet. GaOx som produceras i skakkolvsproduktion visar ett lovande utbyte med en betydande specifik aktivitet mot galaktosen som substrat och kan användas i framtiden för enzymatisk uppgradering av träets biomassa. / Wood is the best renewable source for producing many products due to its biocompatible and biodegradable nature. The wood biomass has a recalcitrance nature towards enzymatic upgrading. It is due to various reasons such as lignin content, acetyl content in hemicellulose, and cellulose crystallinity which blocks the enzyme binding site. This study comes under BioUPGRADE, which is a collaborative platform to create high-value and multipurpose materials sustainably using biocatalysis. The general aim of this study is to produce holocellulose and hemicellulose substrates from different wood species and produce, purify, and validate galactose oxidase as a potential biocatalyst for wood fiber modification. The study investigates the effect of chemical peracetic acid delignification on two wood species, eucalyptus hardwood (HW), and spruce softwood (SW) investigated at different time intervals, where the PAA was prepared ex-situ with a volumetric ratio of 1:3. With the results from the PAA treatment, 38.53% of lignin was removed in eucalyptus and 31.80% in softwood. The hemicellulose yield increased by 47.40% for eucalyptus and 19.05% for spruce with an increase in the time of PAA treatment. The acetyl content of the hemicellulose was reduced from 2% to 0.6% in hardwood and 1.96% to 0.6% in softwood. The cellulose recovery after the delignification was nearly 100%. The galactose oxidase was produced in a shake flask using the Pichia pastoris KM71H strain. Pichia pastoris KM71H cells were cultivated using the buffered complex glycerol media (BMGY) and galactose oxidase was expressed using the Pichia pastoris KM71H strain in Buffered complex methanol media (BMMY). The expressed GaOx protein was subsequently purified using AKTA chromatography using a 5ml Histrap FF column. To determine the protein concentration Bicinchoninic acid (BCA) analysis was performed and the GaOx produced in shake flask cultivation was 286.25 mg/L. The specific activity of the shake flask produced GaOx is 164.24 U/mg. It can be observed that PAA treatment proves to be an efficient method for delignification as the cellulose recovery is near 100% and the loss of hemicellulose is relatively low with the volumetric ratio used. The GaOx produced in shake flask production shows a promising yield with a significant specific activity towards the galactose as substrate and could be used in the future for the enzymatic upgrading of the wood biomass.

Soil organic matter (SOM) contains about twice the amount of carbon in the atmosphere. With global changes, the potential shifts in SOM quantity and quality are a major concern. Due to its heterogeneity, SOM remains largely unknown in terms of its molecular composition and responses to climatic events. Traditional bulk soil analysis cannot depict the structural changes in SOM. This thesis applies two complementary molecular-level methods, i.e., SOM biomarker gas chromatography/mass spectrometry (GC/MS) and nuclear magnetic resonance (NMR) spectroscopy, to examine the origin and degradation of various SOM components in grassland and temperate forest soils, and to investigate the shifts in microbial community and SOM composition with both laboratory- and field-simulated global changes, such as frequent freeze-thaw cycles, increasing soil temperatures, elevated atmospheric CO2 levels, and nitrogen (N) deposition. This thesis has several major findings. First, as the most active component in soil, microbial communities were sensitive to substrate availability changes resulting from prolonged soil incubation, freeze-thaw-induced cell lyses, N fertilization and increased plant inputs under elevated CO2 or soil warming. Microbial community shifts have direct impacts on SOM decomposition patterns. For instance, an increased fungal community was believed to contribute to the enhanced lignin oxidation in an in situ soil warming experiment as the primary degrader of lignin in terrestrial environments. Second, contrast to the conventional belief that aromatic structure was recalcitrant and stable in SOM, ester-bond aliphatic lipids primarily originating from plant cutin and suberin were preferentially preserved in the Canadian Prairie grassland soil profiles as compared with lignin-derived phenols. Cutin- and suberin-derived compounds also demonstrated higher stability during soil incubation. With an increased litter production under elevated CO2 or global warming, an enrichment of alkyl structures that had strong contributions from leaf cuticles was observed in the Duke Forest Free Air CO2 Enrichment (FACE) and soil warming experiments, suggesting an accumulation of plant-derived recalcitrant carbon in the soil. These results have significant implications for carbon sequestration and terrestrial biogeochemistry. Overall, this thesis represents the first of its kind to employ comprehensive molecular-level techniques in the investigation of SOM structural alterations under global changes.

Soil organic matter (SOM) contains about twice the amount of carbon in the atmosphere. With global changes, the potential shifts in SOM quantity and quality are a major concern. Due to its heterogeneity, SOM remains largely unknown in terms of its molecular composition and responses to climatic events. Traditional bulk soil analysis cannot depict the structural changes in SOM. This thesis applies two complementary molecular-level methods, i.e., SOM biomarker gas chromatography/mass spectrometry (GC/MS) and nuclear magnetic resonance (NMR) spectroscopy, to examine the origin and degradation of various SOM components in grassland and temperate forest soils, and to investigate the shifts in microbial community and SOM composition with both laboratory- and field-simulated global changes, such as frequent freeze-thaw cycles, increasing soil temperatures, elevated atmospheric CO2 levels, and nitrogen (N) deposition. This thesis has several major findings. First, as the most active component in soil, microbial communities were sensitive to substrate availability changes resulting from prolonged soil incubation, freeze-thaw-induced cell lyses, N fertilization and increased plant inputs under elevated CO2 or soil warming. Microbial community shifts have direct impacts on SOM decomposition patterns. For instance, an increased fungal community was believed to contribute to the enhanced lignin oxidation in an in situ soil warming experiment as the primary degrader of lignin in terrestrial environments. Second, contrast to the conventional belief that aromatic structure was recalcitrant and stable in SOM, ester-bond aliphatic lipids primarily originating from plant cutin and suberin were preferentially preserved in the Canadian Prairie grassland soil profiles as compared with lignin-derived phenols. Cutin- and suberin-derived compounds also demonstrated higher stability during soil incubation. With an increased litter production under elevated CO2 or global warming, an enrichment of alkyl structures that had strong contributions from leaf cuticles was observed in the Duke Forest Free Air CO2 Enrichment (FACE) and soil warming experiments, suggesting an accumulation of plant-derived recalcitrant carbon in the soil. These results have significant implications for carbon sequestration and terrestrial biogeochemistry. Overall, this thesis represents the first of its kind to employ comprehensive molecular-level techniques in the investigation of SOM structural alterations under global changes.