O lançamento de compostos nitrogenados em corpos d’água pode resultar em problemas ambientais, sendo assim tratar águas residuárias, como o esgoto sanitário com o objetivo de remover não apenas matéria orgânica, mas tambémnitrogênio tem sido objeto de estudo há algumas décadas. O objetivo deste trabalho foi avaliar o desempenho de um reator de leito estruturado e fluxo contínuo, com recirculação, na remoção de matéria orgânica e nitrogênio presente em esgoto sanitário, sob diferentes ciclos de Aeração intermitente (AI) e avaliar a influência destes ciclos no desenvolvimento de bactérias nitrificantes (Bactérias Oxidadoras de Amônia – BOA e Bactérias Oxidadoras de Nitrito – BON) e desnitrificantes (DESN) aderidas (Material Suporte - MS) e em suspensão (Efluente – EF e Lodo – LD). O reator utilizado possuía volume útil de 9,4 L. Como material suporte (MS) foi utilizado espuma de poliuretano, cortada e fixada em hastes de PVC. Foram trabalhadas 3 fases de aeração (AE) e não aeração (AN) com diferentes tempos: Fase 1 (4 h AE/ 2 h AN); Fase 2 (2 h AE/ 1 h AN) e Fase 3 (2 h AE/ 2 h AN). Durante todas as fases o tempo de detenção hidráulico foi mantido em 16 h e o efluente recirculado a uma vazão de 3 vezes a vazão afluente. Foram analisados: pH, alcalinidade total, temperatura, Demanda Química de Oxigênio (DQO), Demanda Bioquímica de Oxigênio (DBO), Nitrogênio Kjeldhl Total (NKT), N-amoniacal – N-NH4+, nitrito- N-NO2 e nitrato- N-NO3. A concentração das BOA, BON e DESN foi determinada utilizando o Número Mais Provável.gSSV-1 (NMP.gSSVmL-1). Na Fase 1 o percentual de remoção de NKT, N-NH4+ e NT foi de 76±10%, 70±21% e 67±10% respectivamente. Na Fase 2 de 80±15% de remoção de NKT, 86±15% de N-NH4+ e 68±9% de remoção de NT e na Fase 3 de 58±20%, 72±28% e 41±6% de NKT, N-NH4+ e NT, respectivamente. A eficiência de desnitrificação nas 3 fases foi acima de 70%, indicando que no reator ocorreu o processo de Nitrificação e desnitrificação Simultânea (NDS). Os percentuais de remoção de DQOT foram de 88%±4 na Fase 1, 94%±7 na Fase 2 e 90%±11 na Fase 3. A ANOVA multivariada aplicada ao NMP.gSSV-1, indicou que houve diferença significativa (F: 20,2; p-valor <0,01) entre a concentração dos organismos analisados nos diferentes ciclos de AI. Porém as diferenças entre os NMP.gSSV-1 não depende apenas de fatores isolados e sim de quais meios, fases e grupos estão sendo analisados. Dos resultados conclui-se que o sistema foi eficiente em termos de remoção de nitrogênio e matéria orgânica, e que a fase que apresentou a maior disponibilidade de Oxigênio Dissolvido (OD) e relação C/N (Fase 2), foi a que obteve as menores concentrações efluentes de matéria orgânica e N-NH4+. Apontasse que houve diferença significativa entre a concentração (NMP.gSSV-1) dos organismos analisados (BOA, BON e DESN), porém esta diferença não depende de fatores isolados e sim de quais meios (MS, EF ou LD, fases (1, 2 ou 3) e grupos (BOA, BON e DESN) está sendo considerado. / The release of nitrogen compounds in water bodies can result in many environmental problems, so treat wastewater, such as sewage in order to remove not only organic matter but also nitrogen has been studied a few decades. From the above, the objective of this study was to evaluate the performance of a structured bed reactor, continuous flow, with recirculation, in removing organic matter and nitrogen present in wastewater under different cycles of intermittent aeration (AI) and to evaluate the influence of these cycles in the development of nitrifying bacteria (Oxidizing Bacteria Ammonia - BOA and Bacteria Oxidizing Nitrite - BON) and denitrifying (DESN) adhered (Support Material - MS) and suspension (Effluent - EF and sludge - LD). The reactor used has usable volume of 9.4 L. As support materials (MS) polyurethane foam was used, cut and fixed in PVC rods. 3 were worked aeration phases (AE) and non-aeration (AN) at different stage: Stage 1 (4 h EA / AN 2H); Stage 2 (2H EA / AN 1 h) and Phase 3 (2H EA / AN 2 h). During all hydraulic detention time phases was kept at 16 h and the effluent recirculated at a rate of 3 times the inflow. Were analyzed: pH, total alkalinity, temperature, chemical oxygen demand (COD), Biochemical Oxygen Demand (BOD), nitrogen Kjeldhl Total (NKT), ammonia-N-N-NH4+, nitrito-N-NO2+andnitrato-NO3-. The concentration of BOA, BON and DESN was determined using the number More Provável.gSSV-1 (NMP.gSSV-1). In phase 1 the percentage removal NTK N-NH4+ and NT was 76±10%, 70±21% and 67±10% respectively. In Phase 2 80±15% of removel NKT, 86±15% of N-NH4+ e 68±9% of removel NT e na Fase 3 de 58±20%, 72±28% and 41±6% of NKT, N-NH4+ of NT, respectively. The denitrification efficiency in stage 3 was over 70%, indicating that occurred in the reactor the process of simultaneous nitrification and denitrification (NDS). DQOT the removal percentages were 88 ± 4% in Phase 1, 94 ± 7 in Phase 2 and 90± 11% in Phase 3. The multivariate ANOVA applied to NMP.gSSV-1, it indicated that there was significant (F: 20,2, p <0,01) between the analyzed concentration of organisms AI in different cycles, but the differences between NMP.gSSV-1 depends not only isolated factors but of which means, and phase groups being analysis. From the results it is concluded that the working system is efficient in terms of nitrogen removal and organic matter, and that the stage with the highest availability of Dissolved Oxygen (DO) and C/N ratio (Step 2), was the one obtained the lower concentrations of organic matter effluents and N-NH4+. Hinted that there was a significant difference between the concentration (NMP.100mL-1) of the analyzed organizations (BOA, BON and DESN), but this difference does not depend on factors alone but of which means (MS, EF or LD), stages (1, 2 or 3) and groups (BOA, BON and DESN) is being considered.

Water and wastewater treatment solutions incorporating biofilm systems are becoming increasingly popular due to more stringent regulations pertaining to drinking water and wastewater effluent discharge in Canada and in other parts of the world. As a major component of biofilm, extracellular polymeric substances (EPS) have been considered as an important factor affecting the physical and chemical properties of biofilm. Further, the selected method of EPS extraction and the methods of detecting the composition of the EPS have shown to affect the results of EPS measurements. In this research, protocols for EPS extraction and EPS composition analysis were investigated and optimized for nitrifying moving bed biofilm reactor (MBBR) biofilm. In addition, the confocal Raman microscopy (CRM) spectra of EPS in nitrifying MBBR biofilm and the protein, polysaccharide and extracellular DNA (eDNA) percent concentrations of the EPS were investigated at various operating temperatures. Further, the CRM spectra and the protein, polysaccharide and eDNA percent concentration of EPS in nitrifying MBBR biofilm along with the biofilm morphology and thickness and the viability of the embedded cells were investigated at various hydraulic retention times (HRTs). The EPS was characterized at various temperatures and HRTs in order to investigate potential correlation between the EPS components of the nitrifying biofilm and the ammonia removal kinetics. The biofilm morphology and thickness along with the bacterial viability of the biofilm were also investigated at various HRTs. Biofilm morphology images and thickness measurements were acquired using a variable pressure scanning electron microscope (VPSEM). The percentages of viable embedded cells in the biofilm were quantified using live/dead staining in combination with confocal laser microscopy (CLSM) imaging. The research demonstrates that an increase in protein content and subsequently a decrease in polysaccharides and eDNA contents in the EPS of nitrifying MBBR biofilm were observed at the lowest operational HRT and the highest temperature in this work. In particular, the EPS protein to polysaccharide (PN/PS) ratio of nitrifying MBBR systems was shown to significantly decrease below a value of 3 when the system was underloaded (observed at the highest operational temperature in this study) or hydraulically overloaded (observed at the lowest HRT in this study). As such, data obtained at lower operational temperatures, with the system no longer underloaded, and at longer HRTs, with the system no longer hydraulically overloaded, all demonstrate an EPS PN/PS ratio of approximately 3. Correlations were observed between the chemically measured EPS PN/PS ratios and the measured Raman spectra intensity ratios; supporting the concept of higher PN/PS ratios of EPS in more optimal nitrifying MBBR operations. Further, the ammonia removal kinetics and EPS response at HRT values of 0.75 and 1.0 h indicate that nitrifying MBBR systems may be optimized to operate at HRTs as low as 0.75 to 1.0 hour as opposed to conventional HRTs of 2.0 to 6.0 h.

Metalaxyl, [N—(2,6-Dimethylphenyl)-N-(Methoxyacetyl)-alanine methyl ester], is used extensively in tobacco (Nicotiana tabacum L.) production for prevention of black shank (Phytophthora parasitica Dast. var. nicotianae), blue mold (Peronospora tabacina Adam), and damping-off (Pythigm spp.). Metalaxyl is also patented as a nitrification inhibitor, although not marketed for that purpose. Proper maturity and ripening of flue-cured tobacco depends on an adequate supply of N through the time of removal of the inflorescence, with a declining supply of N from that point. Use of a chemical which might prolong the availability of N in tobacco could delay maturity and reduce the quality of the cured leaf. These studies were conducted to determine whether metalaxyl might inhibit nitrification under a broad range of soil physical and environmental conditions prevalent in the tobacco producing areas of Virginia. The influence of soil type, soil pH, soil temperature, and soil moisture on inhibition of nitrification by metalaxyl (1 mg kg⁻¹) were investigated in three soils used extensively for tobacco production. Soils used in the study were Cecil sandy loam (clayey, kaolinitic, thermic Typic Hapludult), Appomattox fine sandy loam (clayey, mixed, thermic Typic Kandhapludult), and Mattoponi sandy loam (clayey, mixed, thermic Typic Hapludult). Metalaxyl did not inhibit nitrification under any of the conditions studied. However, NO₂⁻ accumulation with metalaxyl was sometimes greater than the control, especially at high pH (7.0) in the Cecil and Appomattox soils, and at 10 and 20°C. Nitrite and NO₃⁻ accumulations from four rates of metalaxyl (1, 5, 25, and 125 mg kg⁻¹) were compared with those of an untreated control and a nitrapyrin standard over a seven week soil incubation period in further studies using the same soils and adjusted pH levels. Significant NO₂⁻ accumulation occurred during the first week after treatment at high pH in all soil types, with 5, 25, and 125 mg kg⁻¹ metalaxyl. Only the 125 mg kg⁻¹ metalaxyl treatment caused NO₂⁻ accumulation at the high pH in all soils beyond the second week after treatment, with the peak occurring in most cases between weeks three and four. Nitrate accumulation proceeded normally in all soil types and pH levels except with treatments of 25 and 125 mg kg". Nitrate accumulations with 25 mg kg⁻¹ were similar to those for nitrapyrin. The 125 mg kg⁻¹ rate was consistent in causing near total inhibition of NO₃⁻ accumulation at all pH levels in all soils. Nitrate accumulation tended to be lower at lower soil pH levels compared to the highest pH for all soils. Little difference in nitrification due to soil appears to be evident. Use of metalaxyl at recommended rates of 0.25 to 1.5 mg kg⁻¹ would not be expected to inhibit nitrification. / Ph. D.

Anaerobic chemolithotrophic growth of Rhizobium japonicum occurred in the presence of carbon monoxide with nitrate serving as the electron acceptor. Under conditions where the atmospheric concentrations of CO were varied; the cells grew in up to 50% (v/v) CO. Optimum growth was observed in the presence of 20-30% CO. Under these conditions absorbance (660 nm) reached a maximum of 0.33 after 15 days of growth, after which no further increase was noted. Colony forming units increased from 1 X 10⁶ cells per ml to a maximum of 2.0 X 10⁸ cells per ml of culture medium. Growth in the absence of CO was substantially less indicating that CO was required for growth. The rate of disappearance of NO₃⁻-nitrogen from the culture medium during growth was correlated with the growth rate. Hydrogen uptake was measured amperometrically with cells grown in the presence of CO with nitrate, nitrite, nitrous oxide or oxygen serving as the electron acceptor. Addition of acetylene, in the presence of N₂O resulted in a 92.5% inhibition of N₂O-dependent H₂ uptake. Demonstration of H₂ uptake activity with NO₃⁻, NO₂⁻ or N₂O as the only electron acceptor substantiated the presence of dissimilatory nitrate, nitrite, and nitrous oxide reductase(s). Hydrogenase activity with O₂ as the electron acceptor in CO grown cultures showed the presence of cytochrome components necessary for transferring electrons from H₂ to O₂. Increasing the H₂ concentration above 0.5% (v/v) resulted in repression of growth in CO grown cultures. CO:oxidoreductase, the enzyme responsible for oxidation of CO was demonstrated in anaerobic, CO grown cultures. / M.S.

The newly recognized ammonia-oxidizing archaea (AOA) makes re-evaluation of the contribution to ammonia oxidization by both AOA and ammonia-oxidizing bacteria (AOB) necessary and meaningful. The growing population and increasing anthropogenic activities around coastlines have affected wetland and coastal marine ecosystems through discharging polluted water containing large amounts of reactive inorganic nitrogen. The objectives of this study were to detect the phylogenetic diversity and abundance of ammonia oxidizers including AOA and AOB on different scales and to elucidate the distribution patterns along an anthropogenic pollution gradient from the coastal wetland of the Mai Po Nature Reserve in Hong Kong to the South China Sea (SCS). Generally, besides lineages shared by similar environments, various endemic lineages were also observed in the polluted mangrove sediments of Hong Kong, and in the coastal, and deep-sea surface and subsurface sediments from the SCS indicating their geographical distance should be responsible for these phylogenetic distinctions. The community structures of AOA and AOB observed were proposed to be associated with environmental parameters including metals and total phosphorus (TP) separately in the sediments while their abundance was correlated with the pH value and temperature. On the other hand, along a profile of surface sediments with stable salinity from the coastal margin to the slope in the SCS, a clear community structure transition was detected for both AOA and AOB, showing major differences in each of their responses. Although the abundance of AOA was lower than that of AOB in the subsurface sediment samples from the SCS, the statistical support for relationships between AOA and nitrite concentration shed new light on the active contributor to the subsurface nitrogen cycle in the oxygen minimum zone from the deep-sea sediments. On a large scale, along the anthropogenic pollution gradient from the Pearl River Delta to the coastal margin and then the SCS, the dominant genus transition from Nitrosomonas to Nitrosospira was detected in response to the salinity and anthropogenic influences. Among a wide spectrum of environmental conditions in the western Pacific, a suite of statistical analyses clearly delineated the shallow and deep-sea sediments clusters suggesting that the depth and other contributing environmental factors involved shape the current distribution pattern of AOA. On a global scale, our understanding about the systematics and evolution of AOA was advanced through phylogenetic analyses. Salinity, lifestyle and temperature were proposed to be responsible for the global distribution patterns of AOA. On the basis of studies in the anthropogenic influence areas, the methods to detect specific responses of ammonia oxidizers to known anthropogenic pollution were concluded. Highlights of this study advance not only our understandings about phylogenetic diversity of ammonia oxidizers and the driving forces shaping their community structure and distribution patterns, but also a revised comprehensive view about them on the larger scale. / published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy

No description available.

Azospirillum brasilense reduced nitrate in W-containing Mo-limited medium, but at lower rates than in W-free medium. However nitrate reduction by Paracoccus denitrificans was completely suppressed under the same conditions. Nitrite reductase activity of growing A. brasilense was negatively affected by tungstate. Nitrite accumulation was shown to be the result of an inhibitory effect of nitrate on nitrite reductase activity. Both resting whole cell and cell-free extract preformed nitrite reductase activities were equally affected by increasing levels of tungstate. Preformed nitrate reductase activity of the cell-free extract was shown to be more sensitive to increasing concentrations of tungstate than whole cell activity, suggesting that the cytoplasmic membrane served as a protective barrier against tungsten inactivation of nitrate reductase.

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.

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.

Orientador: Carlos Alexandre Costa Crusciol / Coorientador: Marcelo de Almeida Silva / Banca: Gabriela Ferraz Siqueira / Banca: Eduardo Negrisoli / Resumo: A cana-de-açúcar (Saccharum spp.) é uma cultura de grande importância econômica para o Brasil, pois além de gerar empregos em setores como agrícola, industriais e terceiros, a cultura também proporciona a produção de biomassa energética. Há muitas evidências de que esta cultura seja beneficiada pela interação com bactérias diazotróficas, principalmente as do gênero Azospirillum que, além da fixação de N atmosférico podem produzir fitormônios que promovem, na maioria dos casos, efeitos positivos no crescimento vegetal, no rendimento e nas alterações fisiológicas da planta. A principal barreira à utilização do Azospirillumn a cultura da cana-de-açúcar tem sido a incon¬sistência dos resultados de pesquisa, que podem variar de acordo com a cultivar, as condições eda¬foclimáticas e a metodologia de condução da pes¬quisa. Objetivou-se, mediante o presente estudo, avaliar a eficácia dos modos de inoculação de Azospirillum brasilense, nos parâmetros biométricos, fisiológicos e tecnológicos, e as possíveis melhorias na absorção de nutrientes do solo e na produtividade. O trabalho foi desenvolvido em área experimental pertencente à Usina da Barra, localizada no município de Santa Maria da Serra - SP, na safra 2015/2016. O delineamento experimental foi em blocos ao acaso, em esquema fatorial 2 x 2, com 4 repetições. Os tratamentos foram constituídos por inoculação do Azospirillum brasilense (sem e com) e pelos modos de aplicação (tolete e foliar). Foi considerada como área útil as 4 ... / Abstract: The sugarcane (Saccharum spp.) is a crop of great economic importance to Brazil, as well as create jobs in several sectors such as agricultural, industrial and third culture also provides the production of energy biomass. There are many evidences that this crop culture is benefited by interaction with diazotrophs, especially the Azospirillum genus that in addition to atmospheric N fixation can produce phytohormones that promote, in most cases, positive effects on plant growth, yield and amendments physiological plant. The main barrier to the use of Azospirillum in sugarcane has been the inconsistency of search results, which may vary according to the cultivar, the climatic conditions and methodology of research. The aim, through this study, was to evaluate the effectiveness of Azospirillum brasilense inoculation modes, the biometric, physiological and technological parameters, and possible improvements in the uptake of soil nutrients and yield. The study was conducted in the experimental area of Sugar mill Barra, localized in Santa Maria da Serra - SP, in 2015/2016. The experimental design was a randomized block in factorial 2 x 2, with four repetitions. The treatments consisted of inoculation of Azospirillum brasilense (with and without) and the application modes (stem and leaf). It was considered useble area of the center lines 4 of 8 lines planted. The inoculation was performed by applying 2 L-1 ha of the commercial product. The inoculation of the stems was performed before covering the plating furrows and in plots that received foliar it occurred at tillering stage. It was concluded that inoculation with Azospirillum brasilense did not affect the macronutrient content, leaf chlorophyll content, stalk number, internode number and the technological quality of sugarcane and there was an increase in stalk height due to the increase in average length internode, resulting in higher yield in ... / Mestre