La contamination des aquifères de proche subsurface par les intrants d'origine agricole (nitrates) est un problème mondial.L'utilisation excessive d'engrais depuis plusieurs décennies a impacté la qualité des masses d'eau souterraines et soulève des enjeux pour la santé humaine comme pour celle des écosystèmes. Les nitrates dans les aquifères peuvent être réduits en diazote gazeux par l'activité microbienne hétérotrophique (la biomasse microbienne obtenant l'énergie nécessaire à ce processus via le carbone organique issu de la surface) et/ou par l'activité autotrophique (la biomasse microbienne obtenant cette fois ci son énergie depuis une source proche, lithologique). Les taux de dénitrification sont très variables spatialement, et sont régulés par l'interaction entre la structure des flux d'eau souterrains avec l'activité biogéochimique. Localiser l'activité biogéochimique dans les aquifères est difficilement réalisable à l'échelle des bassins versants, mais paraît crucial pour la gestion des masses d'eau souterraines. Bien que les processus de l'activité microbienne ne puissent pas être entièrement résolus à l'échelle locale, ce manuscrit de thèse propose une caractérisation des taux de dénitrification à l'échelle du bassin versant, basée sur l'analyse de données et sur une approche de modélisation intégrée. Cette thèse propose d'utiliser de manière extensive des traceurs conservatifs et réactifs associés aux flux d'eau souterraine et des modèles de transport afin d'identifier les contrôles géologiques et biogéochimiques sur les capacités de dénitrification dans les aquifères. Cette méthodologie a été appliquée à un aquifère libre cristallin de 76 km² situé en Bretagne. A partir des concentrations en CFC-12, O2, NO3- et N2 dissous mesurées dans 16 puits, il a été possible de reconstituer les chroniques d'apports de nitrate dans la zone saturée et de définir les variations spatio-temporelles de la dénitrification. Il est prouvé ici que la dénitrification est en premier lieu contrôlée par la position des donneurs d'électron. Ce travail propose un cadre d'interprétation général sur la base de l'utilisation combinée et complémentaire des traceurs et sur la modélisation semi-explicite pour estimer à l'échelle régionale les capacités de dénitrification et les stocks de nitrates dans les aquifères. / Unconfined shallow aquifers in agricultural areas are contaminated by nitrates worldwide. Excessive fertilization over the last decades has affected groundwater quality as well as human and ecosystem wellbeing. Nitrate in groundwater can be microbially reduced to dinitrogen gas by heterotrophic (microbes obtaining their energy from surface-derived organic carbon) and autotrophic (microbes obtaining their energy from a lithological source) processes. However, denitrification rates are highly spatially variable, following involved interactions between groundwater flow structures and biogeochemical activity. The location of biogeochemical activity in the aquifer is difficult to access at the catchment scale, but of vast importance to gain predictive capabilities for groundwater management. Even though microbial processes cannot be resolved at the local scale, this dissertation proposes a catchment scale characterization of denitrification rates based on an integrated model- and data-driven approach. The dissertation proposes an extensive use of conservative and reactive tracers combined with groundwater flow and transport models to identify the geological and biogeochemical controls on aquifer denitrification capacities. The methodology is applied to a crystalline unconfined aquifer of 76 km2 size in Brittany, France. Based on CFC-12, O2, NO3-, and dissolved N2 concentrations measured in 16 wells, it is possible to reconstruct historical nitrate inputs to the saturated zone and to define spatiotemporal denitrification activity. It is shown that denitrification is primarily controlled by the location of electron donors. The dissertation proposes a general interpretation framework based on tracer information combined with complementary semi-explicit lumped parameter models to assess regional denitrification capacities and nitrate legacy.

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V současnosti musí velké energetické podniky k dodržení emisních předpisů, zejména pak vyžadovaných limitů NOx, uplatňovat denitrifikační metody. Tématem předkládané diplomové práce jsou moderních denitrifikační metody a jejich praktické uplatnění v lokálních poměrech uhelného kotle s parním výkonem 640 t.h-1 v elektrárně Počerady. Práce obsahuje rešerši moderních denitrifikačních metod používaných velkými uhelnými kotli se zaměřením zejména na sekundární denitrifikační metody. Jsou uvažovány dvě možné varianty denitrifikace – varianta 1 využívá selektivní katalytickou redukci (SCR) a varianta 2 selektivní nekatalytickou redukci (SNCR) společně s nízkoemisními hořáky a stupňovaným přívodem spalovacího vzduchu. Pro výběr vhodné denitrifikační metody jsou studovány investiční náklady jednotlivých variant – nižší investiční náklady (o 19.4%) slibuje varianta 2. Při srovnávání SCR se SNCR vyšlo najevo, že investiční náklady metody SNCR jsou 5krát nižší než metody SCR. V souladu s investičními náklady, s dispozicí kotle a se složitostí jeho instalace je pro navazující studium problematiky využita varianta 2. Stěžejní část práce se zabývá stanovením optimálního tzv. teplotního okna pro konkrétní metodu SNCR. Těžištěm práce je tepelný výpočet ohniště a části deskového přehříváku pro stanovený rozsah paliv a výkon kotle v rozmezí 60-100%. S uvažováním výsledků z výpočtu jsou navrženy dvě vstřikovací roviny, které mají zaručit vysokou efektivitu denitrifikačního procesu při uvažovaných provozních podmínkách kotle. Diplomová práce rovněž diskutuje obecnou vhodnost instalace SNCR a SCR ve stávajících uhelných kotlích.

Nitrogen cycling processes can be tracked using quantitative Polymerase Chain Reaction (qPCR) to determine the presence and qReverse Transcriptase-PCR (qRT-PCR) to determine expression of key genes, or ‘biological markers’, for nitrogen metabolism. Nitrification is catalyzed in part, by two enzymes: ammonia monooxygenase (AMO; NH3 NH2OH) and nitrite oxidoreductase (NXR; NO2- NO3-). For denitrification, four enzymes act sequentially: nitrate reductase (NAR/NAP; NO3- NO2-), nitrite reductase (NIR; NO2- NO), nitric oxide reductase (NOR; NO  N2O), and nitrous oxide reductase (NOS; N2O  N2). A principle of wastewater treatment (WWT) is to remove excess nitrogen by taking advantage of natural nitrogen cycling or biological nitrogen removal (BNR). This process involves using microorganisms to bring influent ammonia through nitrification and denitrification to release nitrogen gas, which does not contribute to eutrophication. A novel shortcut nitrogen removal configuration could increase nitrogen removal efficiency by promoting nitritation/denitritation, reducing the classic nitrogen cycle by removing the redundant oxidation/reduction step to nitrate (NO3-). Here, three nitrogen transformations were used to track the three main phases in the nitrogen cycle; ammonia monooxygenase for nitrification, nitrite oxidoreductase for shortcut, and nitrous oxide reductase for denitrification. Primers for qPCR and qRT-PCR were designed to capture as much sequence diversity as possible for each step. Genes from bacteria known to perform the nitrogen transformations of interest (amoA, nxrB, nosZ) were used to BLAST-query the Integrated Microbial Genomes & Microbiomes database (img.jgi.doe.gov) to find homologs from organisms commonly found in WWT. These sequences were then aligned to find regions sufficiently conserved for primer design. These PCR primers were tested against standards for each gene and used to track nitrogen transformation potential and expression in a novel lab-scale algal photo-sequencing batch reactor which promotes shortcut nitrogen removal from wastewater across three solids retention times (SRT, or mean cell residence time); 5, 10 and 15 days. SRT 15 had the greatest total nitrogen removal with nitritation and denitritation observed. Nitrate was not detected in the first cycle and shortcut nitrogen removal was supported by low levels of nxrB genes and transcripts. Simultaneous nitrification/denitrification was supported by elevated concentrations of nosZ during the light period and less nitrite produced than ammonium consumed. Nitritation was predominantly performed by Betaproteobacteria amoA and nitrous oxide reduction was predominantly from nosZ group I (Proteobacteria-type).

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Throughout Central Florida surface water and ground water are decreasing in quantity and quality in part because of excess Nitrate and Phosphorus nutrients. Stormwater runoff serves as a medium for transport of Nitrate and Phosphorus to surface water and ground water. The goal of this experiment is assess the Nitrate and Phosphorus removal in stormwater using select media. The results of a literature search, batch test experimentation and column test experimentation are used to determine an optimal media blend that may be implemented in detention ponds to reduce Nitrate and Phosphorus. The extensive literature search revealed 32 different media that may be used to remove Nitrate and Phosphorus. Each potential media was qualitatively and quantitatively evaluated based on 5 criteria: 1) relevance, 2) permeability, 3) cost, 4) availability in Florida, and 5) additional environmental benefit. The top 7 performing media: Florida peat, sandy loam, woodchips, crushed oyster shell; crushed limestone, tire crumb and sawdust were selected for batch test experimentation. The aerobic conditions in batch test experimentation prohibited the growth of denitrifying bacteria, therefore media mixes were selected for column test experimentation based on Ammonia and Orthophosphate concentrations. Batch test experimentation showed the most effective media to be 50% sand, 30% tire crumb, 20% sawdust by weight (media mix 1) and 50% sand, 25% sawdust, 15% tire crumb, 10% limestone by weight (media mix 2). Media mix 1, media mix 2 and a control are tested in column test experimentation, where the control is site soil from Hunters Trace development in Ocala, Florida. Column test experimentation models a dry detention pond where water passes through a 48 inch unsaturated zone then a 48 inch saturated zone. To test Nitrate and Orthophosphate removal potential, pond water augmented with Nitrate (0.38, 1.26, 2.5 mg/L NO3-N) and Orthophosphate (0.125, 0.361, 0.785 mg/L PO4-P) was pumped into the columns. Media mix 1 and media mix 2 outperformed the control in both Nitrate and Orthophosphate removal. Media mix 1 and media mix 2 had Nitrate removal efficiencies ranging from 60% to 99% and the control had Nitrate removal efficiencies ranging from 38%-80%. Media mix 1 and media mix 2 averaged Orthophosphate removal efficiencies ranging from approximately 42% to 67%. For every run in every influent Orthophosphate concentration the saturated control added Orthophosphate to the water. The Nitrate and Orthophosphate removal performances for media mix 1 and media mix 2 could not be directly compared because of different influent saturated nutrient concentrations.

Nitrogen removal technologies can help dairy industries meet their nutrient management needs. This thesis investigated two separate treatment technologies for total ammonia nitrogen (TAN) removal: algae raceway ponds and aerated biofilm reactors. Six 1000- liter algae raceway ponds and four 1000-liter tote tanks, each equipped with 10 sheets of nonwoven geotextile (i.e., thermally bonded or needle-punched) biofilm substrate, were used to treat the effluent from a flush dairy in central coastal California (TAN = 251 mg/L, cBOD5 = 204 mg/L). For the algae raceway ponds (TAN loading rate = 7 - 35 g/m³-day among 7-, 10- and 14-day hydraulic residence times (HRT)), first-order removal rate constants (k) were ~0.2 day⁻¹ in the summer and 0.1 - 0.2 day⁻¹ in the winter. Removal rate constants had no correlations (R² < 0.1) with water temperature, weak to moderate (for 7-day ponds, R² = 0.55) correlations with insolation and weak to no correlations with biomass (i.e. volatile solids) concentration. During the winter, low insolation likely inhibited algal photosynthesis and biological TAN treatment. Ponds with 7-day HRT had distinct absence of nitrate and nitrite compared to 10- and 14-day ponds. Net productivities were ~20 g/m²-day in summer and 9 – 11 g/m²-day in winter; gross productivities were 120 – 160 g/m²-day in summer and 77 – 150 g/m²-day in winter. Productivities had no correlations (R² < 0.1) with water temperature and weak to moderate correlations (for 14-day ponds, net productivity R² = 0.56, gross productivity R² = 0.83) with insolation. Analysis of organic compounds in pond effluent showed dissolved volatile solids (~2300 mg/L) were mostly non-biodegradable (~98% of soluble oxygen demand). Dissolved organic nitrogen concentrations in the pond effluent were ~35 mg/L. For the aerated biofilm reactors, tanks with needle-punched geotextiles had greater first- order TAN removal rate constants (0.69 day⁻¹) than tanks with thermally bonded geotextiles (0.23 day⁻¹) while operating in batch mode. Needle-punched geotextile reactor also accumulated sludge faster and had higher attached to water column biomass (i.e. volatile solids) ratios (~0.08 g VS/g VS) than thermally bonded geotextile reactor (~0.04 g VS/g VS). Among the four tanks, mass of attached biomass was 150 – 340 g per tank while mass of biomass in the water column was 3290 – 5430 g per tank. Comparing the two treatment technologies, aerated biofilm reactors (removal = 64 – 77%, k = 0.2 – 0.3 day⁻¹, removal rate = 36 – 43 g-N/m²-day, 16 – 19 g-N/m³-day) had more removal and faster removal rates per square meter of land footprint compared to the algae raceway ponds (removal = 38 – 77%, k = 0.1 – 0.2 day⁻¹, removal rate = 4 – 5 g- N/m²-day, 13 – 17 g-N/m³-day), likely due to direct application of aerators in the reactors.