In this study, we have created a simulation model which is concerned about digesting cellulose, as a major component of microalgae in a bioreactor. This model is designed to generate a computational model that simulates the process of granulation in anaerobic sludge and aims to investigate scenarios of possible granular bioaugmentation. Once a mature granule is formed, pro- tein is used as an alternative substrate that will be supplied to a mature granule. Protein, being a main component of cyanobacteria, will promote growth and incorporation of a cell type that can degrade protein (selective pressure). The model developed in a cDynoMiCs simulation environment successfully demonstrated the process of granule formation and bioaugmentation in an Anaerobic granule. Bioaugmentation is a common strategy in the field of wastewater treatment, used to in- troduce a new metabolic capability to either aerobic or anaerobic granules. The end product of our work is a model that can visually demonstrate varying stratifications of different trophic microbial groups that will be of help for the engineers and researchers, who are operating both laboratory and industrial-scale anaerobic digesters and wish to enhance reactor performance. The working model that we have developed has been validated using the existing literature and lab experiments. The model successfully demonstrates granulation in a cellobiose fed system with formation of 0.63 mm mature granule in 59 days with the production of good amount of methane that could be used commercially as a green fuel. This model is extended to perform bioaugmentation by chaining different simulations.

Aqua, a commercial product manufactured by BiOWiSH® Technologies, was utilized in this research to study its effectiveness to biodegrade methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA). Microcosms containing varying concentrations of MTBE and TBA as well as a growth media and mineral salt solution were examined. Analytical instrumentation used in this study included the use of a gas chromatograph-mass spectrometer (GC/MS) to determine concentrations of MTBE and TBA and a spectrophotometer to extrapolate approximate active biomass concentrations in each experiment. Four different environmental conditions were tested for both MTBE and TBA. The environmental conditions tested for each contaminant included: biodegradation under aerobic conditions, biodegradation under anaerobic conditions, biodegradation under denitrifying conditions, and biodegradation under aerobic conditions with glucose present. This study concluded that there is potential for degradation of MTBE and TBA using Aqua under the conditions tested. Maximum MTBE biodegradation was observed under aerobic conditions which yielded a first order rate constant of 0.019/hour and a 99.8 percent decrease in MTBE over 14 days. Maximum TBA biodegradation was observed under aerobic conditions with glucose present which yielded a first rate order constant of 0.009/hour and a 95.03 percent decrease in TBA concentrations over 14 days. It is presumed that under both conditions a monooxygenase enzymatic reaction involving Cytochrome P-450 aids in breaking down both MTBE and TBA. However, the results presented are indicative of biodegradation under lab conditions with little to no interference. Further research is needed to determine the effectiveness of Aqua utilizing groundwater or soil samples from MTBE or TBA contaminated sites in order to truly analyze Aqua’s potential to be used as a bioaugmentation product in real world applications.

No description available.

Chlorinated solvents are among the most prevalent contaminants at Superfund sites. Perchloroethylene (PCE) and its degradative byproducts pose a particular problem because of their persistence in the subsurface and their threat to ecological health. In this study, microcosms were used to test the viability of bioaugmentation as a possible remediation strategy at a PCE contaminated site at the Naval Amphibious Base at Little Creek located in Virginia Beach, Virginia. All microcosms were created in duplicate using spatially diverse soils and the bioaugmented series innoculated with a mixed microbial culture provided by the Dr. Frank Loffler. This culture has been found to be capable of completely degrading PCE to ethene. The aqueous ethene concentration was monitored over time. It is clear from the results that bioaugmentation successfully increased the degree of reductive dechlorination over their static counterpart. Without innoculation, shallow static microcosms showed an accumulation of cis-DCE, while deep soils never showed conversion beyond TCE. Shallow bioaugmented microcosms showed the production and loss of vinyl chloride indicated probable complete conversion of PCE to ethene while deep soils showed the production of cis-DCE. These differences in dechlorination between shallow and deep soils indicate a possible disparity in reduction capacity. At day 78, microcosms were spiked with higher concentrations of PCE resulting in a reduction in dechlorination activity. Static microcosms exhibited similar degradative trends but bioaugmented batches experienced dramatic reductions in dechlorination activity indicating possible inhibition effects of native organisms due to concentration or potential toxic shock. It appears that bioaugmentation is a remediation alternative worthy of further study including possible delivery methods, toxicity or inhibition effects of concentration, and fate/transport studies. / Master of Science

Les traitements phytosanitaires intensifs en viticulture, conjugués aux flux de ruissellement, se traduisent par l'arrivée de cuivre dans les bassins d'orage (BO). Des phénomènes de bioatténuation impliquant la microflore et les macrophytes s'observent, mais les niveaux d'épuration sont souvent faibles et variables. La phytoextraction du Cu est une technique intéressante in situ mais sa lenteur la pénalise. La bioaugmentation couplée à la phytoremédiation est une solution pour augmenter la vitesse d'extraction. L'étude a consisté à sélectionner un triplet «bactérie-plante-sorbant » afin d'extraire le Cu du sédiment et de réduire sa teneur dans les eaux en sortie de BO. Les trois composantes ont ensuite été assemblées dans des microcosmes remplis d'un mélange sable-sédiment reproduisant les caractéristiques d'un BO de référence. Sur 564 isolats bactériens cultivables, 84 ont montré une capacité à complexer les métaux et l'isolat n°l06 a été retenu pour sa capacité à complexer le Cu. Des trois macrophytes testés, Phragmites australis, espèce retenue dans la suite de l'étude, a accumulé le plus de Cu dans les parties aériennes. Enfin, la pulpe de betterave s'est révélée être le sorbant le plus efficace dans une matrice complexe et en présence d'herbicides. Des expériences en microcosmes, mettant enjeu le triplet sélectionné, ont été réalisées. La localisation de la pulpe de betterave en sortie de microcosmes a réduit de 20% les quantités de Cu exportées par les eaux en atteignant un abattement de 95%. La phytoextraction du Cu a été augmentée par 1,7 grâce à la bioaugmentation dans le cas d'un régime hydraulique semi-continu et d'une inoculation répétée de l'isolat n°106. / Intensive pesticides use in viticulture combined with runoff often result in amounts of Cu getting to storm basins (SB). Biological pesticide mitigation has been observed and related to the role of natural macrophyte together with their associated microflora. Nevertheless, low and variable mitigation levels are most often reported. Cu phytoextraction is a promising in situ technology but requires time. Bioaugmentation-assisted phytoextraction is a new technology that may enhance the phytoextraction rate. The study consisted in selecting the best "microorganism-plant-sorbent" association in order to extract Cu from sediments and mitigate Cu load in SB outlet waters. The association of these three selected components has been studied in SB-scaled microcosms filled with a sand-sediment mixture to a better understanding of the depollution process feasability. Among the 564 isolates extracted from SB sediments, 84 were able to complex metals and isolate 106 was selected for the process implementation on the base of its ability to complex Cu. On the 3 studied macrophytes, Phragmites australis accumulated more Cu in aerial parts. Finally, sugar beet pulp has been selected as the best Cu sorbant in mixture with herbicides and in a complex matrix. Microcosm experiments that associated the best three components were carried out. Results showed that sugar beet pulp located at the outlet of microcosms reduced significantly (20 %) Cu amounts in outlet waters to reach a mitigation rate of 95%. Cu extraction in aerial parts of P. australis has been improved by a factor of 1,7 thanks to an intermittent wate Joad, with drying and rewetting periods along with repeated inoculations.

Cette étude vise à développer un système de dépollution d'eau et de sédiments viticoles. Le glyphosate, diuron et 3,4-dichloroaniline (3,4-DCA) sont considérés, en tenant compte de la charge en cuivre. L'augmentation du temps de séjour des polluants dans le système et le choix de la bioaugmentation couplée à la phytoremédiation est la stratégie retenue. Chaque composante d'un triplet « sorbant-inoculum bactérien-plante » a été sélectionnée en laboratoire et le triplet mis en oeuvre en microcosmes en conditions partiellement contrôlées.La sélection de matériaux sorbants dans différentes matrices liquides en présence des polluants seuls ou en mélange montre que les capacités de sorption des différents matériaux testés varient selon le polluant, sa formulation (seul ou en mélange) et la matrice liquide. Le sédiment se révèle être le meilleur sorbant pour le glyphosate ; le sable pour le diuron et le 3,4-DCA.Les performances de dissipation des colonies tolérantes isolées varient fortement selon le polluant. Le consortium sélectionné pour le procédé dissipe le glyphosate, le diuron et le 3,4-DCA en milieu liquide et complexe par ailleurs le cuivre.L'expérience en microcosmes montre que le temps de rétention hydraulique influe sur l'efficacité du système et que les matériaux sorbants sont efficaces. De plus les performances de dissipation atteintes sont bonnes. L'effet de la bioaugmentation sur les performances de dissipation n'est pas significatif pour le glyphosate et le 3,4-DCA, mais améliore en moyenne la dissipation du diuron. Un temps d'action prolongé dans la matrice solide s'avère nécessaire pour obtenir une bonne efficacité du procédé. / This study aims at developing a depollution system for vineyard sediments and passing water. Glyphosate, diuron and 3,4-dichloroaniline (3,4-DCA) have been chosen, taking a copper load into account. Strategical choices are increased pollutant hydraulic retention time and the use of bioaugmentation together with phytoextraction. Each item on this « sorbent-bacterial inoculum-plant » triplet has been beforehand selected in the laboratory and implemented in microcosm experiments under semi-controlled conditions.Sorbent material selection in different liquid matrices with pollutants alone or in combination shows that material sorption capacities change with the type of pollutant. It has also been shown that these capacities vary with formulation (alone or in combination) and liquid matrix chosen. Sediment is awarded the best sorbent for glyphosate, sand the best sorbent for diuron and 3,4-DCA.Dissipation performances from isolated colonies are strongly related to the nature of the compound. Elected consortium degrades glyphosate, diuron and 3,4-dichloroaniline (3,4-DCA) in liquid culture as well as it complexes copper.Microcosm experiments show that hydraulic retention time weighs on pollutant concentration in water at the system outlet. Sorbents prove to be efficient. Dissipation performances are satisfactory. Bioaugmentation does not impact on glyphosate and 3,4-DCA dissipation performances significantly, but enhances diuron dissipation in average. Suitable process efficiency requires increased action time within the solid matrix.

Les résidus pharmaceutiques sont considérés comme un problème écologique émergent, à cause de leur présence et leur accumulation continue dans l’environnement. Même à des faibles concentrations, ces substances sont susceptibles de menacer l’ensemble des organismes vivants. Il est donc, urgent de développer les moyens techniques permettant leur élimination. Dans ce cadre s’inscrit le travail de cette thèse, il a pour objectif de traiter la carbamazépine, un antiépileptique largement détecté dans le milieu aquatique. Une première étude a été menée sur la biodégradation de la carbamazépine par des boues activées par biostimulation avec différentes sources de carbone conventionnelles. Une optimisation des différents paramètres opératoires a été également effectuée. En outre, une seconde étude a porté sur la biodégradation de la molécule cible par une souche fongique, et ce travail a été achevé par une troisième étude de synthèse mettant en place la technique de bioaugmentation fongique des boues activées afin d’améliorer la performance du traitement. / Pharmaceutical residues are considered an emerging environmental problem because of their presence and their continuous accumulation in the environment. Even at low concentrations, these substances may threaten all living organisms. It is therefore urgent to develop the technical means to eliminate them. In this framework is the work of this thesis, it intended to treat carbamazepine, an antiepileptic drug widely detected in the aquatic environment. A first study was conducted on the biodegradation of carbamazepine by activated sludge by biostimulation with various sources of conventional carbon. An optimization of the various operating parameters was also performed. In addition, a second study examined the biodegradation of the target molecule by a fungal strain, and this work was completed by a third synthesis study establishing technical fungal bioaugmentation of activated sludge to improve performance treatment.

Anurans are declining worldwide because of the spread of Batachochytrium dendrobatidis (Bd), the fungus that causes chytridiomycosis. However, some frogs are resistant to this disease, and understanding why may be critical to saving those that are susceptible. In Peru, Gastrotheca excubitor is resistant to chytridiomycosis while Gastrotheca nebulanastes is susceptible. Two anuran skin defenses, symbiotic bacteria and antimicrobial peptides (AMPs), have demonstrated the ability to inhibit Bd in vitro when isolated from certain frogs. We tested if these defenses can explain the difference in susceptibility between the two Gastrotheca species. The cutaneous bacteria and AMPs of both species were collected, tested for their abilities to inhibit the growth of Bd, and analyzed for their compositions. Results indicate that 34%of the strains of skin bacteria from G. excubitor were able to inhibit the growth of Bd whereas only 10% isolated from G. nebulanastes were effective. Gastrotheca excubitor also has stronger anti-Bd skin bacteria. Neither frog species has peptide mixtures capable of completely inhibiting Bd, and overall species did not differ in the anti-Bd abilities of their peptides. These results suggest that the chytridiomycosis resistance experienced by G. excubitor may be attributed to its skin bacteria.

In recent years, due to the continuous development of machinery, electronics, leather, chemical companies and dry-cleaning industry, more and more chlorinated hydrocarbons accumulate in the soil, causing serious harm to the environment. The accumulation of chlorinated hydrocarbons and the teratogenic, carcinogenic, and mutagenic hazards seriously threaten human health. Therefore, the remediation of chlorinated hydrocarbons is imminent. Under this premise, in-situ bioremediation has gradually received attention. For in situ bioremediation of soil contaminated with chlorinated hydrocarbons, the most commonly used methods are biostimulation alone, bioaugmentation alone, and a combination with biostimulation and bioaugmentation. The removal rate of trichloroethylene in the case of using biostimulation products alone is significantly lower than that of using bioaugmentation products alone. The removal rate of trichloroethylene by biostimulation products alone does not exceed 60%, and “DCE pause” occurred, but did not occur in the case of using bioaugmentation products. The removal rate of trichloroethylene by bioaugmentation products is generally higher than 98%, and it will promote the degradation of trichloroethylene or tetrachloroethylene to non-toxic ethylene. Therefore, only cases containing bioaugmentation can achieve non-toxic degradation of chlorinated hydrocarbons and take into account the high removal rate of them. In addition, the biostimulation duration is significantly shorter.

Microbial electrolysis cell (MEC) system is a potential technology that could treat wastewater while simultaneously generating H2 (green energy). MEC's electroactive bacteria (EAB) are essential microbes responsible for oxidizing organic pollutants (such as acetate) in wastewater using an electrogenesis process. Since EABs comprise the core of MECs, they are essential for maintaining functional stability (Coulombic efficiency (CE), current density, and pollutant removal) of MECs. The cause of EAB becoming dominant at the anode of MECs fed with wastewater is still unclear. Furthermore, efficient EAB are typically not detected in wastewater, and when they are present their abundance is low, which affects their early colonization on the anode and subsequent growth into a mature biofilm. This study investigated bioaugmentation as a strategy to drive the assembly of functionally redundant anode EAB biofilms to improve MEC performance. Two bioaugmentation strategies (Conditions 2 and 3) with known EABs (G. sulfurreducens and D. acetexigens) were tested during the startup of MECs. Meanwhile, control MEC reactors (Condition 1) were operated with only wastewater as the sole source of inoculum to compare the anodic biofilm assembly and system performance with the bioaugmented reactors. Equal number of G. sulfurreducens and D. acetexigens cells were added to the wastewater-fed MEC (10% inoculum at 2.1E+07 live cells/mL). In Condition 3, anodic-biofilm colonized G. sulfurreducens and D. acetexigens was used as anode in wastewater fed MECs. Using single-chambered MEC reactors, the bioaugmented MECs (Condition 2 and 3) performed more efficiently than the non-bioaugmented (Condition 1) MECs. Current generation, CE and gas production were different between the three conditions tested (Condition 3 > Condition 2 > Condition 1). Analysis of 16S rRNA gene sequencing of anodic biofilm indicates revealed that the bacterial communities was not affected between the tested conditions. However, the relative abundance of EABs, mainly G. sulfurreducens and D. acetexigens, was markedly influenced by bioaugmentation compared to the control reactor. The highest peak current generation (~ 1500 mA/m2), CE (70.3 ± 9%), and gas production (0.04 m3/m3/day) was observed in Condition 3. Collectively, these results provide a framework for engineering the anode microbial communities in MECs for wastewater treatment.