Na aquacultura de recirculação são utilizados filtros biológicos para o tratamento do efluente antes que este retorne aos tanques. Estas unidades de tratamento têm como finalidade a transformação de nitrogênio amoniacal em nitrato, pois a amônia e o nitrito são tóxicos para os peixes. O nitrogênio amoniacal tende a se acumular na água de cultivo, pois é um importante produto de excreção dos organismos aquáticos e degradação da ração não consumida. Nestes filtros biológicos, os microorganismos responsáveis pelo tratamento da água residuária se encontram aderidos no meio de preenchimento, formando um biofilme. O presente trabalho teve como objetivo caracterizar as comunidades microbianas presentes no biofilme associado ao filtro biológico de uma unidade experimental de tratamento de efluente de aquacultura. Durante o experimento, realizado em dois sistemas paralelos representando unidades de aquacultura com e sem recirculação de água, também foram monitorados parâmetros de qualidade da água. Os tanques experimentais foram povoados com juvenis de tilápias-do-Nilo (Oreochromis niloticus), que foram submetidos a pesagens a cada quinze dias para avaliar seu ganho de biomassa. As bactérias foram identificadas através da técnica de análise microbiológica da hibridização fluorescente in situ (FISH). A estrutura do biofilme foi avaliada através de microscopia eletrônica de varredura. Os resultados indicam que a nitrificação teve um papel mais importante no controle da qualidade da água no sistema com recirculação em relação ao tanque sem recirculação. A análise microbiológica do meio de preenchimento do filtro revelou uma presença marcante (com proporções de Cy3/DAPI variando entre 0,5% e 7,6%) de células ativas de organismos nitrificantes (oxidadores de amônia e de nitrito), pertencendo a gêneros distintos como Nitrobacter, Nitrococcus e Nitrosomonas, além de outros grupos de presença expressiva, como bactérias filamentosas (com proporções de 11,2% a 17,3% da contagem de células marcadas com DAPI). As imagens de Microscopia Eletrônica de Varredura revelaram a natureza do arranjo destas bactérias no meio filtrante, caracterizando um biofilme bem desenvolvido, composto por diversos morfotipos microbianos. O conhecimento das bactérias que compõe este biofilme pode tornar possível a geração de melhorias que podem ser implementadas para aumentar a eficiência do sistema. / In recirculating aquaculture, biological filters are used for treating the effluent before it returns to tanks. These treatment units are intended for transforming ammoniacal nitrogen into nitrate, since ammonia and nitrite are toxic to fish. Ammoniacal nitrogen tends to accumulate in culture water, because it is an important excretion product from aquatic organisms and also due to degradation of non consumed feed. In these biological filters, microorganisms responsible for the treatment of waste water adhere to the filler, forming a biofilm. The present work intended to characterize the microbial community present in the biofilm associated to the biological filter at an experimental aquaculture effluent treatment unit. During the experiment, conducted in two parallel systems representing aquaculture units with and without water recirculation, water quality parameters were also monitored. Experimental tanks were populated with juvenile Nile Tilapias (Oreochromis niloticus), which were subjected to weighing every 15 days in order to assess their biomass gain. Bacteria were identified through the microbiological analysis technique of Fluorescent In Situ Hybridization (FISH). The biofilm structure was assessed using scanning electron microscopy. The results indicate that nitrification had a more important role in the control of water quality in the system with recirculation compared with the tank without recirculation. The microbiological analysis of the filter media revealed a significant presence (with Cy3/DAPI range between 0.5% e 7.6%) of active cells from nitrifying organisms (ammonia and nitrite oxidizers), which belonged to different genera such as Nitrobacter, Nitrococcus and Nitrosomonas, in addition to groups that had an expressive presence, such as filamentous bacteria (representing 11.2% to 17.3% of the total DAPI stained cells). Scanning Electron Microscopy images revealed the nature of the arrangement of these bacteria in the filtering media, characterizing a well developed biofilm made up of diverse microbial morphotypes. The knowledge about the bacteria making up the biofilm may enable improvements that can be implemented to increase system effectiveness.

Legumes play an important role in agriculture as major food sources for humans and as feed for animals. Bioavailable nitrogen is a limiting nutrient for crop growth. Legumes are important because they can form a symbiotic relationship with soil bacteria called rhizobia that results in nitrogen-fixing root nodules. In this symbiosis, rhizobia provide nitrogen to the legumes and the legumes provide carbon sources to the rhizobia. The Medicago truncatula NPF1.7/NIP/LATD gene is essential for root nodule development and also for proper development of root architecture. Work in our lab on the MtNPF1.7/MtNIP/LATD gene has established that it encodes a nitrate transporter and strongly suggests it has another function. Mtnip-1/latd mutants have pleiotropic defects, which are only partially explained by defects in nitrate transport. MtNPF1.7/NIP/LATD is a member of the large and diverse NPF/NRT1(PTR) transporter family. NPF/NRT1(PTR) members have been shown to transport other compounds in addition to nitrate: nitrite, amino acids, di- and tri-peptides, dicarboxylates, auxin, abscisic acid and glucosinolates. In Arabidopsis thaliana, the AtNPF6.3/NRT1.1( CHL1) transporter was shown to transport auxin as well as nitrate. Atchl1 mutants have defects in root architecture, which may be explained by defects in auxin transport and/or nitrate sensing. Considering the pleiotropic phenotypes observed in Mtnip-1/latd mutant plants, it is possible that MtNPF1.7/NIP/LATD could have similar activity as AtNPF6.3/NRT1.1(CHL1). Experimental evidence shows that the MtNPF1.7/NIP/LATD gene is able to restore nitrate-absent responsiveness defects of the Atchl1-5 mutant. The constitutive expression of MtNPF1.7/NIP/LATD gene was able to partially, but not fully restore the wild-type phenotype in the Atchl1-5 mutant line in response to auxin and cytokinin. The constitutive expression of MtNPF1.7/NIP/LATD gene affects the lateral root density of wild-type Col-0 plants differently in response to IAA in the presence of high (1mM) or low (0.1 mM) nitrate. MtNPF1.7/NIP/LATD gene expression is not regulated by nitrate at the concentrations tested and MtNPF1.7/NIP/LATD does not regulate the nitrate-responsive MtNRT2.1 gene. Mtnip-1 plants have an abnormal gravitropic root response implicating an auxin defect. Together with these results, MtNPF1.7/NIP/LATD is associated with nitrate and auxin; however, it does not act in a homologous fashion as AtNPF6.3/NRT1.1(CHL1) does in A. thaliana.

No description available.

Submitted in fulfillment for the requirements for the Degree of Doctor of Technology: Biotechnology, Durban University of Technology, 2012. / Nitrification is a crucial step in biological nutrient removal (BNR) processes, mostly carried out by a group of nitrifying bacteria which includes ammonia-oxidising bacteria (AOB) and nitrite-oxidising bacteria (NOB). Nitrification failure has proven to be a common operational problem in full-scale wastewater treatment plants (WWTP) since nitrifying bacteria are very sensitive to sudden changes in environmental or plant operating conditions. The current investigation was carried out to advance our understanding of the distribution of nitrifying bacterial populations and their performance at three different BNR plants in KwaZulu-Natal, South Africa. The latest molecular techniques such as fluorescent in situ hybridisation (FISH)-confocal scanning laser microscopy (CSLM), polymerase chain reaction (PCR) and real-time quantitative PCR (Q-PCR) were applied to detect and quantify nitrifying bacteria. When using FISH to target the nitrifying population, it necessitated optimising pre-treatment protocols of the samples to improve accuracy during quantification. Sonication was found to be the superior method of dispersion based on the least disruption of nitrifier cell integrity, irrespective of the sludge type. The effect of plant configurations and wastewater characteristics on the distribution of the nitrifying bacterial population and subsequently on the nitrification performance was evaluated using FISH and PCR. FISH results revealed the dominance of Nitrosomonas (AOB), Nitrobacter (NOB) and Nitrospira (NOB) for all BNR plants. The 16S rRNA analysis of PCR products using genus-specific primers, revealed the presence of more than one species of the same group at these plants. Nitrosomonas spp. including Nitrosomonas halophila, Nitrosomonas eutropha, Nitrosomonas europaea, Nitrosomonas aestuarii and an unidentified Nitrosomonas spp. were found to dominate among the AOB and Nitrobacter vulgaris, Nitrobacter alkalicus, Nitrobacter hamburgensis and an unidentified Nitrobacter spp. were the dominant species for NOB. Among these species, Nitrosomonas aestuarii, Nitrosomonas europaea, Nitrobacter hamburgensis were detected only from the industrial wastewater samples. The efficiency of two commonly used techniques viz., FISH and Q-PCR for the detection of nitrifiers from WWTP were also studied and compared, specifically targeting Nitrobacter sp. Even though there were slight variations in the quantification results, changes in the Nitrobacter community at these plants were consistent for both FISH and Q-PCR results. Both techniques have their own limitations and advantages. This study has helped to add to the platform of understanding the distribution and activity of nitrifying bacteria by correlating population dynamics with the operational parameters at full-scale level. The observations made in this study will assist researchers and engineers to minimise future nitrification failure at full-scale BNR plants. This study also confirmed the highly complex activities of wastewater treatment processes, which is dependant on a number of factors. Specific AOB or NOB predominant in wastewater rather suggests that the wastewater type and characteristics may contribute to significantly different microbial environments. Among the AOB, Nitrosomonas dominated at all BNR plants throughout the study period and for NOB both Nitrobacter and Nitrospira were found in significant numbers but their dominance varied across the plants. These dissimilar, distinct distribution patterns could be attributed to their environment which in turn impacted on the nitrification performance of the system. It was also noted that the co-existence of more than one group of these communities at the same plant could help the plant escape complete functional failures such as nitrification, due to sudden changes in temperature and substrate concentrations, as this function can be performed by different groups. Although it would have been meritorious to conduct a nitrogen balance in this study, this was not possible since the research focused on full-scale systems. / National Research Foundation / D

Os sistemas de lodos ativados se destacam pelo fato de oferecerem a possibilidade de remoção satisfatória de nutrientes como nitrogênio e fósforo, com poucos requisitos de área. Para aperfeiçoar esse processo de remoção, o sistema pode ser otimizado a fim de alcançar satisfatórias condições de pH, temperatura, aeração, entre outros. As bactérias presentes no lodo ativado são capazes de transformar e remover o nitrogênio através dos processos de nitrificação e desnitrificação. As reações de nitrificação são catalisadas por diferentes gêneros de bactérias autotróficas, entres os quais Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus. Existem evidências mais recentes de que estas reações de nitrificação podem, em certa medida, também ser catalisadas por organismos heterotróficos. Assim, a identificação das bactérias presentes em lodos ativados de Estações de Tratamento de Efluentes (ETE), bem como o estudo dos fatores que podem ou não influenciar na eficiência do processo de remoção de nutrientes, é muito importante para a otimização do sistema. As análises físico-químicas e, posterior, análises da composição bacteriana foram realizadas em amostras de afluente, efluente e de lodo de quatro diferentes estações de tratamento. As coletas foram realizadas em ETE de dois abatedouros de aves, de uma indústria de embalagens de papel e de um hospital particular. As amostras analisadas mostraram resultados pouco similares entre si, evidenciando diferenças nos processos de nitrificação e desnitrificação conforme a ETE de origem. Os resultados encontrados para as análises físico-químicas nas amostras estudadas estiveram de acordo com os parâmetros normalmente encontrados para efluentes industriais e de acordo com os exigidos pela legislação (CONAMA, 2011), com exceção de apenas uma amostra (hospital), que teve um valor de nitrogênio amoniacal (21,23 mg/L) na saída do sistema acima do valor máximo permitido pela legislação vigente (20 mg/L). A eficiência de remoção de DQO em todas as estações de tratamento esteve na faixa entre 67,5% e 97,8%. A amostra da ETE da Indústria de Embalagens de Papel foi a que demonstrou maior eficiência de remoção de DQO (97,8%), enquanto que uma das amostras da ETE do Hospital Particular, que apresentou a maior DQO na entrada do sistema, foi a que obteve menor eficiência de remoção (67,5%). A remoção de nitrogênio amoniacal foi observada, com exceção de uma amostra, em todas as demais, e sua eficiência de remoção foi bastante variável nas amostras, com valores considerados satisfatórios para o parâmetro estudado (99,7%) e abaixo do esperado para sistemas de lodos ativados (24,6%). A menor eficiência de remoção de nitrogênio amoniacal (24,6%) foi encontrada para uma amostra da ETE Hospital, enquanto a maior eficiência se observou para a amostra da ETE Embalagens de Papel (99,7%). As amostras de lodo foram analisadas molecularmente através de nested PCR e DGGE, com posterior sequenciamento genético. Os primers utilizados na técnica de nested PCR foram: 11f e 1492r (primers universais), NIT3r (Nitrobacter), Nos1225r (Bactérias oxidadoras de amônio), Ntspa685r (Nitrospira), sendo o primer Eub338f utilizado em conjunto com os primers específicos. Já o PCR realizado para o DGGE foi realizado utilizando-se os primers 968F GC e 1392r. As análises moleculares demonstraram que as bactérias mais comuns nos processos de tratamento por lodos ativados (Bactérias oxidadoras de amônio, nitrobacter e nitrospira) estiveram presentes (pelo menos um dos gêneros) em quase todas as amostras testadas por PCR. A amostra que mostrou maior eficiência de remoção de DQO e nitrogênio amoniacal teve, também, resultado positivo para todos os primers utilizados (Bactérias oxidadoras de amônio, nitrobacter e nitrospira) na técnica de PCR, sugerindo que o processo de nitrificação foi satisfatório. Das amostras que tiveram resultado positivo para as amplificações com primers específicos para nitrificantes, apenas uma teve correspondência de resultado positivo no sequenciamento genético (Bactérias oxidadoras de amônio). / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2016-11-23T17:48:34Z No. of bitstreams: 1 Dissertacao Carla Maria Pauletti.pdf: 2032023 bytes, checksum: 05c1e6ec8f9dadc338303f4fd9581a5d (MD5) / Made available in DSpace on 2016-11-23T17:48:34Z (GMT). No. of bitstreams: 1 Dissertacao Carla Maria Pauletti.pdf: 2032023 bytes, checksum: 05c1e6ec8f9dadc338303f4fd9581a5d (MD5) Previous issue date: 2016-11-23 / PETROBRAS, Brasil.

Os sistemas de lodos ativados se destacam pelo fato de oferecerem a possibilidade de remoção satisfatória de nutrientes como nitrogênio e fósforo, com poucos requisitos de área. Para aperfeiçoar esse processo de remoção, o sistema pode ser otimizado a fim de alcançar satisfatórias condições de pH, temperatura, aeração, entre outros. As bactérias presentes no lodo ativado são capazes de transformar e remover o nitrogênio através dos processos de nitrificação e desnitrificação. As reações de nitrificação são catalisadas por diferentes gêneros de bactérias autotróficas, entres os quais Nitrosomonas, Nitrobacter, Nitrospira, Nitrococcus. Existem evidências mais recentes de que estas reações de nitrificação podem, em certa medida, também ser catalisadas por organismos heterotróficos. Assim, a identificação das bactérias presentes em lodos ativados de Estações de Tratamento de Efluentes (ETE), bem como o estudo dos fatores que podem ou não influenciar na eficiência do processo de remoção de nutrientes, é muito importante para a otimização do sistema. As análises físico-químicas e, posterior, análises da composição bacteriana foram realizadas em amostras de afluente, efluente e de lodo de quatro diferentes estações de tratamento. As coletas foram realizadas em ETE de dois abatedouros de aves, de uma indústria de embalagens de papel e de um hospital particular. As amostras analisadas mostraram resultados pouco similares entre si, evidenciando diferenças nos processos de nitrificação e desnitrificação conforme a ETE de origem. Os resultados encontrados para as análises físico-químicas nas amostras estudadas estiveram de acordo com os parâmetros normalmente encontrados para efluentes industriais e de acordo com os exigidos pela legislação (CONAMA, 2011), com exceção de apenas uma amostra (hospital), que teve um valor de nitrogênio amoniacal (21,23 mg/L) na saída do sistema acima do valor máximo permitido pela legislação vigente (20 mg/L). A eficiência de remoção de DQO em todas as estações de tratamento esteve na faixa entre 67,5% e 97,8%. A amostra da ETE da Indústria de Embalagens de Papel foi a que demonstrou maior eficiência de remoção de DQO (97,8%), enquanto que uma das amostras da ETE do Hospital Particular, que apresentou a maior DQO na entrada do sistema, foi a que obteve menor eficiência de remoção (67,5%). A remoção de nitrogênio amoniacal foi observada, com exceção de uma amostra, em todas as demais, e sua eficiência de remoção foi bastante variável nas amostras, com valores considerados satisfatórios para o parâmetro estudado (99,7%) e abaixo do esperado para sistemas de lodos ativados (24,6%). A menor eficiência de remoção de nitrogênio amoniacal (24,6%) foi encontrada para uma amostra da ETE Hospital, enquanto a maior eficiência se observou para a amostra da ETE Embalagens de Papel (99,7%). As amostras de lodo foram analisadas molecularmente através de nested PCR e DGGE, com posterior sequenciamento genético. Os primers utilizados na técnica de nested PCR foram: 11f e 1492r (primers universais), NIT3r (Nitrobacter), Nos1225r (Bactérias oxidadoras de amônio), Ntspa685r (Nitrospira), sendo o primer Eub338f utilizado em conjunto com os primers específicos. Já o PCR realizado para o DGGE foi realizado utilizando-se os primers 968F GC e 1392r. As análises moleculares demonstraram que as bactérias mais comuns nos processos de tratamento por lodos ativados (Bactérias oxidadoras de amônio, nitrobacter e nitrospira) estiveram presentes (pelo menos um dos gêneros) em quase todas as amostras testadas por PCR. A amostra que mostrou maior eficiência de remoção de DQO e nitrogênio amoniacal teve, também, resultado positivo para todos os primers utilizados (Bactérias oxidadoras de amônio, nitrobacter e nitrospira) na técnica de PCR, sugerindo que o processo de nitrificação foi satisfatório. Das amostras que tiveram resultado positivo para as amplificações com primers específicos para nitrificantes, apenas uma teve correspondência de resultado positivo no sequenciamento genético (Bactérias oxidadoras de amônio). / PETROBRAS, Brasil.

The soil is a complex system composed by live organisms, organic matter, gases, water and minerals parts that interact. In this sense, the soil organisms, besides inhabitants, are part of its composition. In the soil, the agricultural activity normally begin in a natural area, with several species of plants and animals living in equilibrium, may pass to a reduction of biodiversity, in function of agricultural practices. The present study was carried out in phases, aiming to evaluate and characterize the faunal composition, the population of nitrifying bacteria, and to develop a protocol to extract earthworms DNA. The faunal composition and nitrifying bacteria, population was done in an experiment with five years of no-till with the following treatments: a) witness (without lime); b) amount indicated by SMP method to pH 6.0 (100%), incorporated to soil in 20 cm depth; c) amount indicated by SMP method to pH 6.0 (100%), distributed in soil surface; d) half of the amount indicated by SMP method to pH 6.0 (50%), distributed in soil surface; and e) one quart of the amount indicated by SMP method to pH 6.0 (25%), distributed in soil surface. To evaluate the population of soil meso and macrofauna were collected samples in July (winter) and December (summer) of 2001. For epiedafic meso and macro organisms were installed traps for capture, and for euedafic macro organisms, were collected soil monoliths. The samples collected individually were placed in plastic bags, packed in thermo boxes and taken to Soil Biology Laboratory of UFSM Soils Department. Later, were performed separation, counting and classification of taxonomic groups in level of classes and order of the organisms? Were utilized the Shanon Diversity Index (H) to evaluate the population among the treatments and theTukey test (P=0.05) to compare the means of population in the different doses and mode of lime application. The doses and mode of lime application in no-till after 5 years were not a significant factor to affect the abundance and richness of soil fauna. The most representative population of organisms was Collembola. The quantitative determination of bacteria nitrifying population was performed with seeding of dispersed soil over a layer of silica gel and later addition of a mix composed of lime, alkaline silicates and an acid mixture, that became neutral forming a second layer of silica gel that contains the nutrients specific to the growth of the bacteria group studied. The experiment was performed in triplicates with 25 mg of soil samples per plaque. After 10 days of incubation in 28-30oC were done observations and counting of the colonies developed in each plaque, presuming that each bacteria formed one colony. The counting was done every 5 days for a period of 60 days. The application of 100% lime, incorporated or on soil surface, and 50% of lime on soil surface did not were significantly different in the number of colonies of nitrifying bacteria. The pH under 4.3 in the treatments witness and 25% of lime on soil surface affected negatively the bacteria population in depths 0-5 and 5-10 cm. To develop a protocol to extract DNA of earthworms, earthworms were collected in the experiment of doses and mode of lime application in the no-till system, in the experimental area of UFSM Soils Department. After visual characterization of identical groups, 10 individuals of each specie of Eisenia foetida and Pheretima spp were placed in alcohol 70% for later characterization after morphological patterns of each specie. Other 10 individuals were open and taken all materials from its interior. Before DNA extraction, the material stayed in sterilized water for 24 hours in refrigerator to eliminate any substances that may be stayed glued to the material. Later, the material was macerated and the DNA extracted. The DNA extracted was visualized in agarose gel 1.2%. The protocol developed was efficient to extract the DNA of Eisenia foetida and Pheretima spp. / O solo é um sistema complexo composto de seres vivos, ar, água, matéria orgânica e minerais que interagem. Neste sentido os organismos do solo, além de habitantes, são parte integrante de sua composição. No solo, a atividade agrícola que geralmente inicia como área natural, com muitas espécies de plantas e animais convivendo em equilíbrio, pode passar a uma redução da biodiversidade, decorrentes das práticas culturais. O presente estudo foi realizado em três etapas visando avaliar e caracterizar a composição faunística, a população de bactérias nitrificadoras e desenvolver um protocolo de extração de DNA para oligoquetas. A avaliação faunística e da população de bactérias nitrificadoras foi realizada em um experimento com cinco anos de plantio direto com os seguintes tratamentos: A) testemunha (sem calcário); B) quantidade recomendada pelo método SMP para pH 6,0 (100%), incorporada ao solo na profundidade de 20 cm; C) quantidade recomendada pelo método SMP para pH 6,0 (100%), distribuída na superfície do solo; D) metade da quantidade recomendada pelo método SMP para pH 6,0 (50%), distribuída na superfície do solo; E) um quarto da quantidade recomendada pelo método SMP para pH 6,0 (25%), distribuída na superfície do solo. Para avaliação da população da meso e macrofauna, foram coletadas amostras em julho (inverno) e dezembro (verão) de 2001. Para meso e macro organismos epiedáficos foram instaladas armadilhas de captura, e para macrorganismos euedáficos foram coletados monólitos de solos. As amostras coletadas foram, acondicionadas em caixas de isopor e levadas ao Laboratório de Biologia do Solo do Departamento de Solos da UFSM. Posteriormente, foi realizada a separação, contagem e classificação de grupos taxonômicos em nível de classe e ordem. Utilizou-se o índice de diversidade de Shannon (H) para avaliação das populações e para comparação, as médias nos diferentes tratamentos entre as doses e modos e aplicação de calcário através do teste Tukey a 5%. As doses e modo de aplicação de calcário em sistema de plantio direto após 5 anos foram fatores menos determinantes do que as condições climáticas (inverno-verão) na abundância e riqueza da fauna edáfica. A população de organismo mais representativa foi de colembola. A determinação quantitativa de população de bactérias nitrificadoras foi através da semeadura do solo dispersado sobre uma camada de sílica gel e posterior adição de uma mistura composta por calcário, silicatos alcalinos e uma mistura ácida, que contem os nutrientes específicos para o desenvolvimento do grupo bacteriano em estudo. O experimento foi realizado em plaqueamento triplicado com 25 mg de amostra de solo por placa. Após 10 dias de incubação a 28-30o C foram realizadas observações e contagens das colônias desenvolvidas nas placas, realizadas a cada 5 dias por um período de 60 dias. As aplicações de calcário 100% na superfície, 100% incorporada e 50% na superfície não diferiram significativamente no número de colônias de bactérias nitrificadoras. O pH inferior a 4,3 nos tratamentos 0% e 25% de calcário na superfície diminuiu a população de bactérias nitrificadoras. As oligoquetas, para o trabalho de desenvolvimento de um protocolo de extração de DNA, foram coletadas no experimento citado acima. Após caracterização visual de grupos idênticos, 10 indivíduos de cada espécie de Eisenia foetida e Pheretima sp. foram caracterizadas segundo padrões morfológicos. Outros 10 indivíduos foram abertos e retirado todo o material existente no seu interior. Posteriormente o material foi submetido à maceração e extração do DNA. O DNA extraído foi visualizado em gel de agarose 1,2%. O protocolo desenvolvido foi eficiente para extração de DNA em Eisenia foetida e Pheretima sp.

Sorghum plants excrete phenolic acids which reduce subsequent crop yields. These acids accumulate in field soil by combining with soil and clay particles to form stable complexes which remain until degraded by bacterial metabolism. The amount of phenolic acids in soil samples were obtained by gas chromatography measurements, while Azotobacter populations were obtained by plate counts in 40 sorghum field samples from Denton County, Texas. One can conclude that increasing the Azotobacter population in the soil increased the degradation rate of phenolic acids proportionally. It is proposed that seed inoculation will introduce selected strains of Azotobacter into the soil. The presence of Azotobacter should increase crop size in subsequent plantings.

There is an urgent need to develop ammonia removal treatment systems for municipal and industrial wastewater treatment due to the increasingly stringent ammonia effluent discharge regulations implemented by Canada, the United States, and the European Union. The objective of this dissertation is to develop new understanding and advance the current design and operation of total ammonia nitrogen (TAN) removal via the moving bed biofilm reactor technology (MBBR) for municipal and industrial wastewaters. The first specific objective is to develop a passive, low operationally intensive, efficient and robust design strategy for municipal wastewater treatment to achieve partial nitritation (PN) as a pre-treatment to anammox treatment without using control strategies such as operating at low dissolved oxygen, or the use of inhibitors. This first objective includes developing new knowledge of the biofilm, biomass and microbiome of attached growth PN systems. The second specific objective is to investigate the impact of defining a maximum biofilm thickness, via bio-carrier design, to enhance the effects of free nitrous acid inhibition for PN of municipal wastewaters. The third objective is to investigate the effect of influent copper concentration on nitrifying MBBR systems over long-term operations, to demonstrate the feasibility of the nitrifying MBBR as a solution for TAN removal from gold mining wastewaters. The results pertaining to the first objective, achieved via a study investigating the operation of a nitrifying moving bed biofilm reactor at elevated TAN surface area loading rates (SALRs) of 3, 4, 5, and 6.5 g TAN/m²∙d with the aim of achieving passive PN, demonstrates that operating at a TAN SALR value of 6.5 g TAN/m²∙d can achieve PN without restricting dissolved oxygen or using inhibitors. Operating at a TAN SALR value of 6.5 g TAN/m²∙d achieves a TAN surface area removal rate (SARR) of 3.5 g TAN/m²∙d, and a nitrite accumulation of 99.8% of the oxidized TAN, demonstrating the suppression of nitrite oxidizing bacteria (NOB) activity, while achieving elevated TAN SARR values. At the molecular-scale, there is a statistically significant change in the ammonia oxidizing bacteria (AOB) to NOB ratio from 1:2.6 to 8.7:1 as the TAN SALR increases from 3 to 6.5 g TAN/m²∙d; however, even at a TAN SALR value of 6.5 g TAN/m²∙d there is an NOB abundance of approximately 2%; thus demonstrating that NOB remain present in the biofilm, while their activity is suppressed by operation at elevated TAN SALR values. Furthermore, this system was shown to achieve stable PN consistently for over a period of 10 months of operation, demonstrating a robust, passive, low operational strategy for attached growth PN. The second objective of this dissertation is addressed through a study that compared the carrier design of defined maximal biofilm thickness (z-prototype carrier) to undefined maximal biofilm thickness (chip-prototype carrier) for PN via free nitrous acid inhibition of tertiary, low carbon, municipal wastewaters. The study demonstrates that defined maximal biofilm thickness is a preferred design choice to achieve attached growth PN. The chip-prototype carrier shows biofilm thicknesses and biofilm mass values that are ten-fold higher than the z-prototype carrier, which is shown to contribute to the impact of free nitrous acid on AOB and NOB activities. The z-prototype carrier shows PN is achieved after 3 hours of exposure to free nitrous acid while the chip-prototype carrier does not achieve PN within this same time of exposure. Therefore, the defined maximal biofilm thickness carrier is identified in this research as the preferred design option to achieve attached growth PN for municipal, low carbon, tertiary wastewater treatment. The results of the third objective, achieved via a study investigating the effects of influent copper concentrations on nitrifying MBBR during long term operations to gold mining wastewaters, demonstrates that there is no AOB inhibition in attached growth systems exposed to 0.1, 0.3, 0.45, and 0.6 mg Cu/L for long exposure times. A trend of increasing nitrite accumulation with increasing influent copper concentrations is shown, indicating that NOB inhibition occurs at influent copper concentrations of 0.3 mg Cu/L and greater, with the greatest NOB inhibition observed with an influent copper concentration of 0.6 mg/L. There is no statistically significant difference in biofilm characteristics at the copper concentrations tested; however, there is a trend of increasing biofilm thickness and biofilm roughness with increasing copper concentrations. This study demonstrates the resilience of the nitrifying biofilm to copper inhibition and demonstrates that the nitrifying MBBR is a promising system for removing TAN in mining wastewater in the presence of copper.

Thesis: Ph. D., Joint Program in Oceanography/Applied Ocean Science and Engineering (Massachusetts Institute of Technology, Department of Earth, Atmospheric, and Planetary Sciences; and the Woods Hole Oceanographic Institution), 2019 / Cataloged from PDF version of thesis. / Includes bibliographical references. / Nitrogen (N) species in hydrothermal vent fluids serve as both a nutrient and energy source for the chemosynthetic ecosystems surrounding deep-sea vents. While numerous pathways have been identified in which N-species can be produced and consumed in the context of submarine hydrothermal vent systems, their exact nature has been largely limited to interpretation of variations in concentrations. This thesis applies stable isotope approaches to further constrain the sources and fate of N-species in deep-sea vents across a variety of geological settings. First, I discuss isotope fractionation and reaction kinetics during abiotic reduction of nitrate (NO₃⁻) to ammonium ([sigma]NH₄⁺ = NH₃+NH₄⁺) under hydrothermal conditions. Results of lab experiments conducted at high temperatures and pressures revealed a wide degree of N isotope fractionation as affected by temperature, fluid/rock ratio, and pH-all which exert control over reaction rates. / Moreover, a clear pattern in terms of reaction products can be discerned with the reaction producing [sigma]NH₄⁺ only at high pH, but both [sigma]NH₄⁺ and N₂ at low pH. This challenges previous assumptions that NO₃⁻ is always quantitatively converted to NH₄⁺ during submarine hydrothermal circulation. Next, I report measurements of [sigma]NH₄⁺ concentrations and N isotopic composition ([delta]¹⁵N[subscript NH4]) from vent fluid samples, together with the largest compilation to date of these measurements made from other studies of deep-sea vent systems for comparison. The importance of different processes at sediment-influenced and unsedimented systems are discussed with a focus on how they ultimately yield observed vent [sigma]NH₄⁺ values. / Notable findings include the role that phase separation might play under some conditions and a description of how an unsedimented site from Mid-Cayman Rise with unexpectedly high NH4+ may be uniquely influenced by N₂ reduction to [sigma]NH₄⁺. Lastly, I explore [sigma]NH₄⁺ dynamics in the context of low-temperature vent sites at 9°50'N East Pacific Rise to investigate dynamics of microbially-mediated N transformations. Through both measurements of natural samples, as well as isotopic characterization of N species from incubation experiments and model simulations thereof, an exceptionally high variability observed in [delta]¹⁵N[subscript NH4] values emphasizes the complexity of these microbe-rich systems. / In sum, this thesis highlights the role of microbial processes in low temperature systems, demonstrates a more mechanistic understanding of lesser-understood abiotic N reactions and improves the coverage of available data on deep-sea vent [sigma]NH₄⁺ measurements. / by Chawalit "Net" Charoenpong. / Ph. D. / Ph.D. Joint Program in Oceanography/Applied Ocean Science and Engineering (Massachusetts Institute of Technology, Department of Earth, Atmospheric, and Planetary Sciences; and the Woods Hole Oceanographic Institution)