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

O objetivo deste trabalho foi o de avaliar o desempenho de um reator de leito estruturado submetido à aeração intermitente (LEAI) na remoção de matéria orgânica e nitrogênio de esgoto sanitário. O reator apresentava um volume total de 11,6 L, com uma porosidade do leito de 52%. O leito estruturado era composto de 13 estruturas cilíndricas (diâmetro igual a 3 cm) de espuma de poliuretano, dispostas verticalmente no interior do reator. A alimentação do reator era feita continuamente com esgoto sanitário, sendo este proveniente de um interceptor de esgoto que passava próximo ao laboratório onde o sistema foi montado. O reator era provido de sistema de recirculação interna, com razão de recirculação igual a 3, valor este adotado após a realização de ensaios hidrodinâmicos. Foram testadas 4 condições operacionais diferentes, variando-se o tempo de detenção hidráulica (TDH) e os períodos de aeração e não aeração. Para cada condição operacional, foram feitos os seguintes estudos: avaliação da eficiência de remoção de matéria orgânica e nitrogênio, determinação da atividade nitrificante e desnitrificante, determinação do número mais provável (NMP) da comunidade nitrificante e desnitrificante, perfis temporais e análise da comunidade microbiana. O sistema apresentou elevada remoção de N-total, com valores médios de 80% para valores de TDH de 12 e 10 horas, sob 2 horas de aeração e 1 hora sem aeração. Com a diminuição do TDH para 8 horas, houve queda na eficiência do sistema para 70% (mantendo-se os períodos de aeração de 2 horas e não aeração de 1 hora). Alterando-se o período de aeração para 3 horas, mantendo-se em 1 hora o período sem aeração, o reator apresentou eficiência média de remoção de N-total de 79% com o TDH de 8 horas. A remoção de DQO foi elevada em todas as fases, com valores médios em torno de 90%. Os resultados da atividade nitrificante e desnitrificante foram condizentes com valores encontrados na literatura para sistemas que operam com remoção simultânea de matéria orgânica e nitrogênio. Os perfis temporais mostraram que não houve variação nas formas de nitrogênio (N-NH4+, N-NO2- e N-NO3-) no efluente, apesar da operação sob a aeração intermitente devido ao grau de mistura imposto pela recirculação interna. A concentração de sólidos no efluente foi relativamente baixa quando comparada à concentração de efluentes de reatores de biomassa suspensa, com valor máximo de SSV igual a 108mgSSV.L-1. A presença do Metazoário Aelosomas sp no efluente do reator indica um possível consumo do lodo excedente no reator por esses microrganismos. As análises microbiológicas mostraram uma elevada abundância relativa de gêneros aeróbios facultativos, capazes de utilizar N-NO3- e N-NO2- como receptor de elétrons na ausência de oxigênio. Também se verificou a presença do gênero Comamonas sp em elevada abundância relativa. Esse gênero apresenta microrganismos capazes de realizar a nitrificação heterotrófica e a desnitrificação aeróbia. No entanto, os ensaios realizados não permitem inferir sobre a ocorrência desses processos. Desse modo, os processos convencionais de nitrificação e desnitrificação, tanto pela estratificação do biofilme devido à formação de gradiente de concentração de oxigênio dissolvido, quanto devido à aeração intermitente, certamente tiveram papel preponderante na remoção de nitrogênio. Os resultados obtidos permitem considerar o reator LEAI como uma alternativa tecnicamente viável para a remoção simultânea de matéria orgânica e nitrogênio de esgotos sanitários. / This study aimed at evaluating the organic matter and nitrogen removal from sewage in a structured bed reactor subjected to intermittent aeration. The reactor had a total volume of 11.6 L and bed porosity of 52%. Thirteen polyurethane foam cylinders of 3 cm diameter and 60 cm height were fixed vertically inside the reactor using PVC rods. The reactor was continuously fed with sewage collected from a sanitary sewer passing near the laboratory where the system was assembled. The reactor was provided with internal recirculation system with a recirculation ratio at 3, a value adopted after conducting hydrodynamic tests. Four different operating conditions were tested, varying the hydraulic retention time (HRT) and aeration and non-aeration periods. Nitrogen and organic matter removal efficiencies, nitrifying and denitrifying activities, and the most probable number (MPN) of nitrifying and denitrifying communities were evaluated for each operational condition. The reactor presented a Total-N removal efficiency of 80%, at HRT of 12 hours and 10 hours and aerated and non-aerated periods of 2 hours and 1 hour, respectively. With decreasing the HRT to 8 hours, the Total-N removal efficiency decreased to 70% for the same aeration and non-aeration periods. After increasing the aerated period to 3 hours and keeping the non-aerated periods of 1 hour and HRT of 8 hours, the system attained the Total-N removal efficiency of 79%. COD removal efficiencies were about 90% in all operational condition. Nitrifying and denitrifying activities were similar to those found in the literature for systems presenting simultaneous nitrification and denitrification, and carbon removal. No variation in nitrogen forms (NO3--N, NO2--N and NH4+-N) was observed during a cycle of intermittent aeration. The solids concentration in effluent was relatively low compared to the concentration of suspended-growth biomass reactors, with a maximum value of VSS of 108 mgVSS.L-1. The presence of the metazoan Aelosomas sp in the effluent in all operational conditions indicated a possible use of the excess sludge in the reactor by these microorganisms. Microbiological analysis showed a high relative abundance of aerobic/facultative microorganisms, capable to use NO3--N and NO2--N as electron acceptor in the absence of oxygen. It was also verified the presence of the genus Comamonas sp with high relative abundance. This genus presents microorganisms able to perform heterotrophic nitrification and aerobic denitrification. However, the assays do not allow us to infer the occurrence of these processes. Thus, the conventional processes of nitrification and denitrification certainly played a preponderant role in nitrogen removal both by the stratification of the biofilm due to the formation of a concentration gradient of dissolved oxygen, as due to the intermittent aeration. The results allow considering the reactor as a viable technological alternative for the simultaneous removal of organic matter and nitrogen of sewage.

Made available in DSpace on 2016-04-28T14:59:00Z (GMT). No. of bitstreams: 1 2008 - Debora Alves Gonzaga da Silva1.pdf: 485199 bytes, checksum: 8ad8d9c6be0e3c6fca263c141e2866c3 (MD5) Previous issue date: 2008-09-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / In the objective of advancing the epidemiologic study of the bacterial stain of the tomato, the mechanisms of transmission of Xanthomonas vesicatoria from the plant to the seed and from the seed to the plant have been evaluated. The morphology of seeds and of parts of the seedlings or saplings (tegument, radicles, hypocotyl and cotyledon leaves, and definitive leaves) have also been studied by means of electronic scanning microscopy techniques. The transmission of X. vesicatoria from the plant to the seed has been evaluated upon direct and indirect isolation of parts of ripe fruit (flesh, placenta, placental liquid, water from the first washing of the seeds, entire seeds and ground seeds) in a culture of Agar Nutrient (AN). The fruit were obtained in experimental parcels, conducted at the Horticultural sector of UFRRJ, and inoculated with the phytobacteria through different methods (atomization and injection) combined with different regions of inoculation (flower cluster, flesh, and placenta), during the plant s developing phases (flower, unripened fruit, firm ripe and ripe fruit). The transmission of the phytobacteria from the seeds to the seedlings and saplings was assessed by means of isolating in an AN medium parts of the seedling or sapling (root, tegument, radicles, hypocotyl and cotyledon leaves, and definitive leaves), germinated in different substrata ( germitest paper, sand, and commercialized substrate for saplings). The morphology of seeds and parts of the seedlings was characterized through observations in electron microscope scanning by using samples of fresh seeds (extracted from inoculated fruit), of parts of seedlings or saplings (tegument, radicles, hypocotyl and cotyledon leaves, and definitive leaves) and of seeds inoculated by vacuum procedure at 7, 14, and 21 days after sowing. The treatments applied to the unripened fruit (inoculation by atomization, injection in the placenta and injection in the flesh) were more efficient in the process of transmitting the phytobacteria than the treatment applied to the firm ripe and ripe fruit. It was also observed that the X vesicatoria colonizes the tegument and every part of the seedlings and saplings during the process of germination and emergence. The seed tegument was characterized by an entanglement of trichomes, with a base in the shape of ring and cavity, which may serve as sheltering sites for pathogens, including the X vesicatoria. The process of colonization of tomato seeds by X vesicatoria characterizes itself by the formation of biofilms and fibrils. The presence of stomata colonized by X vesicatoria has been observed in radicles, of seven day old seedlings, and in primary roots originated from the radicles of 20 day old saplings. Few cells of X vesicatoria have been observed on the hypocotyl. The presence of endophytic bacteria forming aggregates with characteristics of an aggressive growth identified as Acinetobacter sp. was detected in many samples of seeds. These same bacteria were detected in many tests of germination and isolation, interfering negatively in the development and recuperation of X vesicatoria in the in vitro tests. / Com o objetivo de se avan?ar no estudo epidemiol?gico da mancha-bacteriana do tomateiro, foram avaliados os mecanismos de transmiss?o de Xanthomonas vesicatoria da planta para a semente e da semente para a planta. Foram feitos, ainda, estudos da morfologia da semente e partes da pl?ntula ou muda (tegumento, rad?cula, hipoc?tilo e folhas cotiledonares e definitivas), atrav?s de t?cnicas de microscopia eletr?nica de varredura. A transmiss?o de X. vesicatoria da planta para a semente foi avaliada a partir de isolamentos diretos e indiretos de partes de frutos maduros (mesocarpo, placenta, l?quido placent?rio, ?gua proveniente da primeira lavagem das sementes, sementes inteiras e sementes trituradas) em meio de cultura Nutriente Agar (NA). Os frutos foram obtidos em parcelas experimentais, conduzidas no setor de Horticultura da UFRRJ, e inoculados com a fitobact?ria por diferentes m?todos (atomiza??o e inje??o) combinados com diferentes regi?es de inocula??o (cacho floral, mesocarpo e placenta), durante as fases de desenvolvimento da planta (flor, frutos verdes, frutos de vez e frutos maduros). A transmiss?o da fitobact?ria das sementes para as pl?ntulas e mudas foi avaliada por isolamentos em meio NA a partir de partes da pl?ntula ou muda (raiz, tegumento, hipoc?tilo, folhas cotiledonares e folhas definitivas), germinadas em diferentes substratos (papel germitest, areia e substrato comercial para mudas). A morfologia das sementes e partes das pl?ntulas foi caracterizada por meio de observa??es em microsc?pio eletr?nico de varredura utilizando-se amostras de sementes frescas (extra?das de frutos inoculados), de partes de pl?ntulas ou mudas (raiz, hipoc?tilo, folha cotiledonar e folha definitiva) e de sementes inoculadas pelo procedimento a v?cuo aos 7, 14 e 21 dias ap?s a semeadura. Os tratamentos aplicados aos frutos verdes (inocula??o por atomiza??o, inje??o na placenta e inje??o no mesocarpo) foram mais eficientes no processo de transmiss?o da fitobact?ria que os tratamentos aplicados aos frutos de vez e maduros. Observou-se, ainda, que X. vesicatoria coloniza o tegumento e todas as partes das pl?ntulas e mudas durante o processo de germina??o e emerg?ncia. O tegumento da semente foi caracterizado por um emaranhado de tricomas, com a base em forma de anel e cavidade, que podem servir como s?tios protetores para pat?genos inclusive X. vesicatoria. O processo de coloniza??o das sementes de tomate por X. vesicatoria se caracteriza pela forma??o de biofilmes e fibrilas. Foram observados em rad?culas, de pl?ntulas com sete dias de idade, e em ra?zes prim?rias, oriundas das rad?culas de mudas com 20 dias de idade, a presen?a de est?matos colonizados por X. vesicatoria. Sobre o hipoc?tilo, foram observadas poucas c?lulas de X. vesicatoria. Em v?rias amostras de sementes foi detectada a presen?a de bact?rias endof?ticas, que formavam agregados com caracter?sticas de crescimento agressivo, identificadas como Acinetobacter sp. Esta mesma bact?ria foi detectada em v?rios testes de germina??o e isolamento interferindo negativamente no desenvolvimento e recupera??o de X. vesicatoria nos testes in vitro.

Candida albicans is an opportunistic pathogen that is present in the normal flora in a majority of individuals. One key factor in C. albicans virulence is the ability to change its morphology from yeast to an elongated or hyphal form. The regulation of this morphogenesis relies in part upon quorum sensing (QS) molecules. C. albicans often exists as part of a mixed culture alongside other microbes and is influenced by their presence as well as the presence of QS molecules that they produce. In this study, a library of diploid homozygous transcriptional regulator knockout (TRKO) mutants were screened to identify strains capable of forming hyphae in the presence of Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli. We identified three strains that showed increased hyphae development compared to wild type C. albicans. The strains identified had deletions of the transcriptional regulating genes Orf19.3928, Orf19.2842 (GZF3), and Orf19.3865 (RFX1). These strains were tested for alterations of filamentation in liquid media, and biofilm formation. All three strains showed increased rates of biofilm formation compared to the wild type. Orf19.3928 showed altered response to farnesol, a marked in biofilm formation and no inhibition of filamentation when farnesol was present in liquid media. The GZF3 deletion strain showed enhanced filamentation with all three bacterial species while the RFX1 deletion strain showed increased filamentation only with E. coli and S. aureus. In spent media, GZF3 showed slight increases in filamentation in E. coli and S. aureus while RFX1 had moderate increases in filamentation in E. coli and S. aureus and slight increases with P. aeruginosa.

More so than ever, Multiple Drug Resistant (MDR) bacteria are on the rise due to overuse of antibiotics along with natural selection for adaptations that enhance drug-resistant properties. One particular bacterial family, Enterobacteriaceae, has been problematic, exhibiting several bacterial members that have developed a precipitous resistance to modern antibiotics and are also primary causative agents of nosocomial, or hospital acquired, infections. Citrobacter freundii (CF) and Enterobacter cloacae (ECL) are two species of the Enterobacteriaceae family causing significant medical concern due to their role in producing numerous opportunistic infections such as bacteremia, lower respiratory tract infections, urinary tract infections, and endocarditis. Adding to the difficulty of this situation is the ability of bacteria to produce biofilms. These biofilms are communities of bacteria that exhibit increased resistance to antibiotic treatment and eradication. Previous work in the laboratory of Dr. Fox at ETSU has identified an uncharacterized product secreted by Klebsiella pneumoniae (KP), another member of the Enterobacteriaceae family, which appears to have inhibitory effects toward CF and ECL. The current study was designed to characterize the effects this secreted product has on CF and ECL biofilms. Through a high throughput microtiter plate assay, the effects of this secreted product were examined on CF and ECL phases of biofilm attachment and maturation. Based on our findings, we have concluded that this secreted product can be categorized as a possible bacteriostatic agent against biofilm cell density, biofilm mass, and cell viability for both biofilm phases of attachment and maturation.

Instead of planktonic growth in nature, many species of bacteria form biofilm to survive in harsh conditions. Although many chronic bacterial infections are caused by bacterial species in a biofilm lifestyle, previous research has focused on studying antibiotic resistance in planktonic growth. Here we used a modified MBEC assay, i.e. biofilm growth on pegs, to determine Escherichia coli biofilm inhibitory concentrations (BIC) of ciprofloxacin, streptomycin and rifampicin and to study the minimal selective concentration (MSC) for ciprofloxacin in E. coli biofilm. We could observe high inhibitory concentrations for all antibiotics in the biofilm pre-formed in media without antibiotics compared to the biofilm formed in antibiotics. We also show preliminary result indicating that sublethal concentrations of ciprofloxacin lead to the selection of ciprofloxacin resistant mutants in biofilm and that the selection level is lower than what was observed in planktonic growing E. coli. With more knowledge in how the biofilm formation precedes in different antibiotic settings, the treatment for chronic biofilm infections used today could be evaluated and changed so that the infections could be eradicated.

Un biocapteur à base de fibre optique en verre de chalcogénures, transparente dans un large domaine du moyen infrarouge (MIR), a été développé. Il permet l'enregistrement de spectres MIR par spectroscopie déportée. Le principe de la mesure est basé sur le concept de la spectroscopie par ondes évanescentes. Pour améliorer la sensibilité, les fibres utilisées sont effilées sur quelques centimètres et ont un profil en diamètre de 450-100-450 µm. La zone effilée sera mise en contact avec l'échantillon à analyser. Différents états métaboliques (physiologique ou pathologique) dans le foie et le sérum ont été analysés. Les coupes de foies et les sérums ont été mis directement en contact avec la partie senseur de la fibre, et des spectres MIR ont ainsi été collectés. L'objectif de ces études est d'identifier deux états métaboliques différents (sain et pathologique) d'un même échantillon. Des modifications spectrales peuvent être mises en évidence et reliées à une ou des altération(s) du métabolisme des lipides, glucides et/ou protides. Ces modifications peuvent être des différences d'intensité et/ou des décalages en nombre d'onde de bandes d'absorption. Tous les résultats sont corrélés avec les dosages sanguins et les colorations histologiques des échantillons analysés. Le développement d'un biofilm bactérien a été suivi in situ et en temps réel. Proteus mirabilis est un micro-organisme pathogène, opportuniste des voies urinaires, qui a développé un comportement multi-cellulaires complexe, corrélé dans l'espace et le temps, ce qui lui permet de coloniser de nouvelles surfaces. Durant le processus de différenciation, le changement de l'état végétatif à celui migrant (pathogène) est accompagné par des modifications des constituants membranaires. Les spectres MIR, enregistrés pendant le développement d'un biofilm à P. mirabilis, permettent de détecter en temps réel une contamination de surface, mais aussi les modifications biochimiques des constituants de la membrane bactérienne. Tous les spectres ont été traités par Analyse en Composantes Principales (ACP), d'une part pour une reconnaissance non-supervisée d'une pathologie, et d'autre part, déterminer la distribution spatiale des phénotypes de P. mirabilis au sein du biofilm

Le principal objectif de cette thèse était de mieux comprendre l'importance des Substances Polymériques Extracellulaires (SPE) dans la structuration et la formation des biofilms benthiques ; tout en s'inscrivant dans une étude plus globale des mécanismes écologiques impliqués dans le fonctionnement des vasières intertidales. La mise au point des dosages biochimiques a été effectuée sur le mucilage de l'algue Chaetomorpha aerea et a permis en parallèle de purifier un polysaccharide sulfaté riche en galactose, présentant une activité bactéricide sélective contre la souche Staphylococcus aureus (ATCC 25923). Les études biochimiques et écologiques menées sur les SPE extraits de la vasière charentaise ont ensuite permis de quantifier leur dynamique de production et leur composition, en fonction des conditions environnementales. La présence de désoxy-sucres et d'acides uroniques au sein des SPE capsulaires a laissé supposer que ces fractions jouaient un rôle important dans la formation et le devenir du biofilm microphytobenthique. La dernière partie des travaux a permis de caractériser les propriétés acide/base de Lewis et hydrophile/hydrophobe de la surface de la micro-algue Navicula jeffreyi, impliquée dans la formation de biofilms benthiques, par des méthodes classiques d'analyse. L'utilisation d'une nouvelle méthode, la Chromatographie Gazeuse Inverse (CGI), a permis d'obtenir des résultats intéressants et relativement similaires, confirmant le caractère prometteur de la CGI pour l'étude des propriétés de surface des micro-organismes.

Cells sense the properties of the surrounding environment and convert this information into changes in gene expression. Bacteria are, in contrast to many multi-cellular eukaryotes, remarkable in their ability to cope with rapid environmental changes and to endure harsh and extreme milieus. Previously, control of gene expression was thought to be carried out exclusively by proteins. However, it is now clear that small regulatory RNAs (sRNA) also carry out gene regulatory functions. Bacteria such as E. coli harbor a large class of sRNAs that bind to mRNAs to alter translation and/or mRNA stability. By identifying mRNAs that are targeted by sRNAs, my studies have broadened the understanding of the mechanisms that underlie sRNA-dependent gene regulation, and have shed light on the impact that this type of regulation has on bacterial physiology. Control of gene expression often relies on the interplay of many regulators. This interplay is exemplified by our discovery of mutual regulation between the sRNA MicF and the globally acting transcription factor Lrp. Through double negative feedback, these two regulators respond to nutrient availability in the environment which results in reprogramming of downstream gene expression. We have also shown that both the transcription factor CsgD, and the anti-sigma factor FlgM, are repressed by the two sRNAs OmrA and OmrB, suggesting that these sRNAs are important players in the complex regulation that allow bacteria to switch between motility and sessility. Bacterial populations of genetically identical individuals show phenotypic variations when switching to the sessile state due to bistability in gene expression. While bistability has previously been demonstrated to arise from stochastic fluctuations in transcription, our results suggest that bistability possibly may arise from sRNA-dependent regulatory events also on the post-transcriptional level.

Face au phénomène de multi-résistance aux antibiotiques des souches de Pseudomonas aeruginosa chez les patients atteints de mucoviscidose, de nouvelles approches doivent être envisagées. L'utilisation des bactériophages pour cibler les bactéries semble être l'une des plus prometteuses. L'efficacité de la phagothérapie semble démontrée par son utilisation en Europe de l'Est depuis des décennies et par les récents résultats obtenus sur des modèles expérimentaux. Cependant, la possibilité de son utilisation chez des patients atteints de mucoviscidose n'a pas encore fait l'objet d'études approfondies. Nous avons démontré l'efficacité des bactériophages in vivo, lors d'une infection pulmonaire létale provoquée par une souche clinique de P. aeruginosa, mais aussi in vitro, pour réduire un biofilm formé par P. aeruginosa. Nous avons aussi étudié la réponse inflammatoire induite par les bactériophages, dans différents modèles in vitro et in vivo, qui s'est révélée quasiment négligeable. Nous avons également mis au point la technique de mesure de différence de potentiel nasale chez la souris pour étudier le transport ionique transépithélial, paramètre fondamental de la mucoviscidose. Les mesures obtenues en présence de bactériophages ne diffèrent pas significativement par rapport aux normes préalablement définies. Enfin, nous avons mis au point une méthode permettant d'évaluer la capacité des bactériophages à infecter des bactéries au sein d'expectorations de patients atteints de mucoviscidose. Nous apportons ainsi des résultats scientifiques concrets qui permettront de mieux appréhender les conditions nécessaires au développement de futurs essais cliniques chez ces patients.