Made available in DSpace on 2014-06-11T19:27:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-08-13Bitstream added on 2014-06-13T18:31:27Z : No. of bitstreams: 1 oliveirafilho_jh_me_jabo.pdf: 697900 bytes, checksum: 8506e7c314e6d9e7dc607082ecfa14a5 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Na produção de cachaça, a microbiota contaminante afeta diretamente o processo fermentativo, originando fermentações indesejáveis, contribuindo para menores rendimentos em álcool e um desbalanceamento na formação dos compostos secundários que caracterizam a bebida. Portanto, são necessárias práticas para minimizar e controlar essas contaminações, sendo a aplicação de antimicrobianos uma alternativa eficaz para tal controle. Dentro desse enfoque, o objetivo deste trabalho foi avaliar a eficiência de antimicrobianos naturais e sua interação com antibióticos convencionais no controle da contaminação bacteriana na produção cachaça. Para as análises tecnológicas e microbiológicas do vinho e pé-de-cuba, utilizou-se o delineamento inteiramente casualizado em parcelas sub-subdivididas, em esquema fatorial 5x2x10 com três repetições, sendo quatro biocidas (extrato de própolis marrom, extrato de própolis verde, ampicilina e interação própolis/ampicilina (P/A)) e controle (sem adição de antimicrobiano), combinando-se os tratamentos em início e final de safra por 10 ciclos fermentativos. Para as análises tecnológicas do mosto, utilizou-se o delineamento inteiramente casualizado em parcelas subdivididas, em esquema fatorial 2x10 com três repetições, sendo duas épocas da safra (inicio e final) por 10 ciclos fermentativos. Foi observado um comportamento distinto entre épocas, com maiores valores de contaminantes e acidez em mosto no final da safra. Dentre os tratamentos, o extrato de própolis marrom, própolis verde, ampicilina e interação (P/A) mantiveram os índices de viabilidade celulares a níveis elevados e superiores a 90%, com menores índices observados para o tratamento controle. As concentrações de bactérias no pé-de-cuba foram significativamente menores com o emprego dos biocidas. No tratamento controle a acidez total do vinho foi superior... / During the cachaça production, the contaminant microbiota affects directly the fermentative process, originating undesirable fermentations, minimizing the alcohol production as well an unbalancing in the secondary compounds formation that characterizes the beverage. Therefore, practices that minimize those contaminations are necessary and the antimicrobial use is one of the efficient alternatives the control. This work aimed to evaluate the natural antimicrobials efficiency as well its interaction with conventional antibiotics to control the bacterial contamination in the cachaça production. A completely randomized design in sub-divided parcels in a factorial 5x2x10 scheme with 3 replications was used, being four biocides (brown propolis extract, green propolis extract, ampicillin and propolis/ampicillin interaction – P/A) and control, combining them in the beginning and final of harvest by 10 fermentative cycles. A different behavior between treatments was observed, being the higher contamination and acidity found in the end of the harvest. Among treatments, the brown propolis extract, green propolis extract, ampicillin and P/A kept cellular viability indexes higher than 90%, and the control treatment was lower. The bacterial concentration in the inoculums was significantly smaller when biocides were used. In the control treatment the residual reducing sugars. The propolis extract and its combination with synthetic antimicrobials were efficient to control contaminant bacteria during the fermentative process

When paper is manufactured, deposits often form in the machines. These deposits are slimelike and can interfere with the papermaking process. The slimelike deposits are aggregates of micro-organisms, also known as biofilm. One single type of micro-organism can form a biofilm, but most biofilms consists of a mixture of several different kinds of micro-organisms and can form on about any conceivable surface. To control the aggregates of micro-organisms a slimecide is added, a so-called biocide. To examine what kind of bacteria that is included in the biofilm and also which bacteria that is killed or not killed by the biocide, Terminal Restriction Fragment Length Polymorphism analysis (T-RFLP) can be used.   In this report we examine biocides impact on biofilm produced in the laboratory.The biocides were first tested for possible interference with the PCR-step of the T-RFLP analysis. None of the tested ten biocides inhibited the PCR process the biofilm was formed on metal plates when these were lowered in a beaker with white water. Three different beakers were set up, one with addition of a biocide with active component 4,5-DICHLORO-1,2-DITHIOLONE from the start, one with the addition of the same biocide after three days and one with no addition at all of biocide. Samples were taken from the beakers and analyzed with T-RFLP.   In this report, we show that biocides affect planktonic and biofilm micro-organisms differently. There are however some micro-organisms in the biofilm that does not get affected by the biocide.   The experimental in this report is a good way of investigate the influence that biocides have on planktonic and biofilm micro-organisms, but to get even greater result the experiment should be done over a longer period of time and repeatedly.

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-04-03T17:25:23Z No. of bitstreams: 1 Oliveira, Hanna Lara da Cruz Dinéas de [Dissertação, 2013].pdf: 1766254 bytes, checksum: 30cf8e6b52cfd32b623ecf94d3cee01b (MD5) / Made available in DSpace on 2017-04-03T17:25:23Z (GMT). No. of bitstreams: 1 Oliveira, Hanna Lara da Cruz Dinéas de [Dissertação, 2013].pdf: 1766254 bytes, checksum: 30cf8e6b52cfd32b623ecf94d3cee01b (MD5) / Os estafilococos coagulase negativos emergem como importantes patógenos nosocomiais, frequentemente isolados em bacteremias humanas, fato intimamente relacionado com o aumento do uso de dispositivos médicos ao longo dos últimos anos. Sua capacidade de aderir a superfícies poliméricas e produzir biofilmes constitui o principal fator de virulência associado a estes dispositivos. A fim de minimizar esta possibilidade, antissépticos são largamente utilizados para desinfecção de pele e mucosas na rotina hospitalar, no entanto, tem-se observado uma diminuição da susceptibilidade bacteriana a estes agentes. A diminuição da susceptibilidade dos microrganismos a antissépticos, associados a sua capacidade de formar biofilmes tem elevado a morbidade, mortalidade, período de internação e os custos relativos ao cuidado com a saúde. No estudo aqui apresentado, foram analisadas 81 cepas de Staphylococcus epidermidis resistentes a meticilina (MRSE) e 55 cepas de Staphylococcus haemolyticus resistentes a meticilina (MRSHa), todas resistentes a meticilina, isoladas de pacientes internados em hospital Universitário localizado na cidade de Niterói-RJ nos anos de 2008 e 2010. A determinação do perfil de susceptibilidade aos biocidas triclosan, clorexidina e cloreto de benzalcônio mostrou que frente ao triclosan, as amostras apresentaram as menores taxas de susceptibilidade bacteriana. Papaína não apresentou atividade antibacteriana. Entretanto, foi capaz de reduzir expressivamente a formação de biofilme (p<0,06) em ambas as espécies, mostrando-se a mais eficiente entre os produtos analisados. Foi capaz também de desintegrar biofilmes maduros formados por Staphylococcus epidermidis. Os biocidas não levaram à redução significante de biofilmes, exceto a clorexidina, que foi capaz de reduzir a formação do biofilme pelas MRSE. Foi possível verificar que cepas de MRSHa tratadas com cloreto de benzalcônio têm 40% menos probabilidade de formar biofilme se comparada às tratadas com triclosan e clorexidina. A análise estatística nos mostrou que a expressão do gene qacA/B influenciou significativamente a formação de biofilme e confirmou a relação existente entre os valores de concentração mínima inibitória (CMI) e a formação de biofilme (p< 0,001) / Coagulase negative staphylococci have emerged as important nosocomial pathogens, commonly isolated in human bacteremia, a fact closely related to the increased use of medical devices over the last few years. Its ability to adhere to polymer surfaces and produce biofilms is the main virulence factor associated with these devices. To minimize this possibility, antiseptics are widely used for disinfection of skin and mucosa in routine hospital, however, there has been an increase in bacterial resistance to these agents. Decreased susceptibility of microorganisms to antiseptics, associated with its ability to form biofilms has high morbidity, mortality, length of stay and costs related to health care. In the study presented here, we analyzed 81 strains of methicillin-resistant Staphylococcus epidermidis (MRSE) and 55 strains of methicillin-resistant Staphylococcus haemolyticus (MRSHa), all meticillin- resistant, isolated from patients hospitalized at University Hospital in the city of Niterói, Rio de Janeiro in 2008 and 2010. The determination of the susceptibility profile of biocides triclosan, chlorhexidine and benzalkonium chloride demonstrated that front of triclosan, the strains have the biggest rates of bacterial susceptibility. Papain showed no antibacterial activity. However, it was able to significantly reduce biofilm formation (p <0.06) in both species, being the most efficient among the analyzed products. It was also able to degrade established biofilms formed by Staphylococcus epidermidis. Biocides showed no significant reduction of biofilms, except chlorhexidine, which was able to reduce biofilm formation by MRSE. We noticed that strains MRSHa treated with benzalkonium chloride are 40% less likely to form biofilm compared to those treated with chlorhexidine and triclosan. Statistical analysis showed that the gene expression qacA/B was significantly influential to biofilm formation and confirmed positive relationship between the values of MIC and biofilm formation (p <0.001)

<p dir="ltr">Cover crops and soil residual herbicides are some of the essential tools within the integrated weed management. When used at cover crop termination, residual herbicides can extend the period of weed control and reduce the selection pressure for herbicide resistance. However, previous research has indicated that the use of cover crops can increase the microbial activity of the soil which, in turn, is the primary route for herbicide degradation in the soil. In addition to the potential effect on the microbial breakdown of herbicides, cover crops can also alter the fate of herbicides by interception. Three field projects were established to (1) investigate the influence of cover crop [cereal rye (<i>Secale cereale</i> L.) and crimson clover (<i>Trifolium incarnatum</i> L.)] use on soil enzyme activities [β-glucosidase (BG) and dehydrogenase (DHA)], its effect on the concentration of residual herbicides (sulfentrazone, <i>s</i>-metolachlor, cloransulam-methyl, atrazine, and mesotrione) in the soil, and the interception of herbicides by cover crop residue; (2) to investigate the effect of cover crop termination strategies (fallow, standing, and roller crimped 1 d prior to application) and simulated rainfall volumes (0, 4.2, and 8.3 mm simulated over 20 min; equivalent to 0, 12.5, and 25 mm h^-1) on atrazine wash off from cereal rye biomass onto the soil; (3) to investigate the effect of cereal rye termination strategies on the concentration of sulfentrazone, <i>s</i>-metolachlor, and cloransulam-methyl in the soil, weed control, and soybean [Glycine max (L.) Merr.] yield.</p><p dir="ltr">Results from the first project suggests that the use of cover crops occasionally resulted in increased BG and DHA activities relative to the fallow treatment. However, even when there was an increase in the activity of these two enzymes, increased degradation of the residual herbicides was not observed. Furthermore, the use of cereal rye or crimson clover as cover crops did not reduce the efficacy of the residual herbicides in controlling weeds early in the growing season.</p><p dir="ltr">In regards to the fate of atrazine after simulated rainfall, the results from the second project indicated that cereal rye biomass accumulation negatively impacted the amount of herbicide reaching the soil at the time of application. Although the roller crimped cereal rye reduced the amount of herbicide reaching the soil relative to the standing cereal rye, it also reduced atrazine leaching below the 0-5 cm of soil. Furthermore, in cover cropping systems with high levels of cereal rye biomass, more than 8.3 mm of rain are required to wash most of the atrazine off of the biomass.</p><p dir="ltr">Lastly, the adoption of the planting green systems resulted in up to 84% interception of residual herbicides by the roller crimped cereal rye biomass. The use of cereal rye as cover crop did not improve the weed control relative to the fallow during the two years of field research. Soybean yield losses ranged from 10 to 44% with the adoption of the planting green system, primarily due to stand losses. However, there are strategies and proper equipment available that can be used to minimize soybean stand losses.</p>

Ces travaux de thèse portent sur l’étude chimique d’espèces marines échantillonnées en Méditerranée. Dans le cadre du projet VALBIM, co-financé par la région PACA et en étroite collaboration avec la société BioPreserv à Grasse mais également avec la Mycoteca Universitatis Taurinensis (MUT) de l’Université de Turin, nous avons isolé et caractérisé 21 molécules dont 14 se sont avérées nouvelles. L’étude de l’algue Caulerpa taxifolia a conduit à la caulerpényne et à quatre nouveaux dérivés de l’acide pyruvique sulfatés. Les travaux sur l’ascidie Polysyncraton sp. ont conduit aux bolascidines A-D, quatre nouveaux métabolites de type bolaamphiphiles dont il n’existe pas à notre connaissance d’équivalent naturel ou synthétique. La première étude chimique de l’éponge Hexadella racovitzai a conduit à deux dérivés de la bromotyrosine, la psammapline A et une nouvelle molécule, la 4-Osulfatocyclobispsammapline A. Par ailleurs, trois nouvelles crambescines C1, conjointement avec une crambescine A2 déjà répertoriée, ont été isolées de Crambe crambe. L’étude du champignon marin Stachybotrys chartarum, isolé de l’éponge Aplysina cavernicola, a conduit à la satratoxine H et à deux nouveau dérivé, la 2,3-dihydrosatratoxine H et l’épi-chartarutine G, ainsi qu’à trois dérivés stachybotrylactame déjà répertoriés dans la bibliographie. Les différents métabolites ont été évalués pour leurs propriétés antibactériennes et cytotoxiques. Ces travaux ont bénéficié de nouvelles approches analytiques (calculs des spectres de dichroïsme circulaire électroniques par TDDFT, réseaux moléculaires) permettant de renforcer l’intérêt des études liées à l’écologie chimique et à la recherche de nouvelles molécules bioactives potentiellement valorisables dans différents domaines. / This PhD work focuses on the chemical study of marine species sampled in the Mediterranean Sea. As part of the VALBIM project, cofinanced by the region PACA and in close collaboration with the SME BioPreserv in Grasse but also with the Mycoteca Universitatis Taurinensis (MUT) of the University of Turin, we have isolated and characterized 21 molecules in total.14 of them were never reported before. The study of the seaweed Caulerpa taxifolia led to caulerpenyne and four new sulphated pyruvic acid derivatives. The work on the ascidian Polysyncraton sp. led to bolascidins A-D, four new metabolites which belong to the bolaamphiphile family, which, to the best of our knowledge, have no natural or synthetic equivalent. The first chemical study of the sponge Hexadella racovitzai led to two bromotyrosine derivatives, psammapline A and a new molecule, 4-O-sulfatocyclobispsammapline A. In addition, three new C1 crambescins, together with one crambescine A2 already reported, have been isolated from Crambe crambe. For all isolated marine invertebrate metabolites, the producer(s) remain(s) to be explored. The study of the marine fungus Stachybotrys chartarum, isolated from the sponge Aplysina cavernicola, led to satratoxin H and three stachybotrylactam derivatives already reported in the literature, and to 2,3-dihydrosatratoxin H and epi-chartarutine G, two new natural products. The different metabolites were evaluated for their antibacterial and cytotoxic properties. This work benefited from new analytical approaches (calculations of electronic circular dichroïsm spectra by TDDFT, molecular networking) and reinforced the interest of studies related to chemical ecology and search for new bioactive molecules to be valorized in different areas.

Contexte: Les agents de nettoyage sont des substances qui aident l'eau dans le processus de nettoyage. Les biocides comprennent les substances utilisées pour désinfecter, désodoriser, stériliser et assainir. L'utilisation d'agents de nettoyage, de biocides et d'autres produits liés au nettoyage est omniprésente. Certaines études suggèrent que l'exposition professionnelle à des substances liées au nettoyage peut être associée au cancer du poumon. Objectif: Examiner l'association entre le risque de cancer du poumon et l'exposition professionnelle aux agents de nettoyage, aux biocides et à d'autres agents de nettoyage. Méthodes: Cette étude utilise les données de deux études cas-témoins basées sur la population sur le cancer du poumon (étude 1: 1979-1986; étude 2: 1996-2001) menées à Montréal. Dans les deux études, les cas comprenaient des hommes ayant reçu un diagnostic de cancer du poumon confirmé histologiquement dans 18 hôpitaux métropolitains de Montréal. Dans les deux études, un ensemble de témoins de population sélectionnés au hasard à partir de la liste électorale du Québec a été établi (étude 2: 762 cas et 899 témoins); tandis que dans l'étude 1, un groupe témoin de cancer supplémentaire a été sélectionné à partir d'un groupe d'autres patients diagnostiqués avec un autre cancer incident (857 cas, 533 témoins de population, 1349 témoins de cancer). Dans les deux études, des antécédents professionnels détaillés ont été recueillis au cours des entretiens; une équipe de chimistes et d'hygiénistes industriels a ensuite évalué l'exposition professionnelle à de nombreuses substances professionnelles, notamment des agents de nettoyage, des biocides, des alcools aliphatiques, de l'ammoniac, de la soude caustique, des cires et des produits de polissage. Une régression logistique multivariée nonconditionnelle a été utilisée pour estimer les ratios des côtes et les intervalles de confiance à 95% du risque de cancer du poumon associé à diverses mesures de l'exposition professionnelle à ces six agents, tout en ajustant pour les facteurs de risque établis. Les interactions selon l'intensité du tabagisme et l'état d'asthme ont été explorées avec l'inclusion de termes de produits croisés. Résultats: Dans l'ensemble, il n'y avait pas d'association cohérente soutenant le rôle de l'exposition professionnelle aux agents de nettoyage, aux biocides et à d'autres agents de nettoyage dans l'étiologie du cancer du poumon. Bien qu'il y ait eu des preuves que l'intensité du tabagisme peut modifier l'association entre la soude caustique et le risque de cancer du poumon dans l'étude 1; où, chez les fumeurs de faible intensité, une augmentation du risque par trois a été observée par rapport à une association nulle observée chez les fumeurs d'intensité moyenne à forte (pInteraction=0,03). Alors que, dans l'étude 2, les personnes exposées professionnellement à des alcools aliphatiques et qui ont déjà souffert d'asthme ont connu quatre fois du risque de cancer du poumon par rapport à une association nulle observée chez ceux qui n'ont jamais eu d'asthme (pInteraction = 0,04). Conclusion: Pris ensemble, les résultats de cette étude ne soutiennent pas le rôle de l'exposition professionnelle aux agents de nettoyage, aux biocides et à d'autres produits de nettoyage dans l'étiologie du cancer du poumon. / Background: Cleaning agents are materials that aid water in the cleaning process. Biocides include materials used to disinfect, deodorize, sterilize, and sanitize. The use of cleaning agents, biocides, and other cleaning-related agents is ubiquitous. Some studies suggest that occupational exposure to cleaning-related substances may be associated with lung cancer. Objective: To examine the association between lung cancer risk and occupational exposure to cleaning-related agents. Methods: This study uses data from two population-based case-control studies on lung cancer (Study 1: 1979-1986; Study 2: 1996-2001) carried out in Montreal. In both studies, cases included men diagnosed with incident histologically confirmed lung cancer identified across 18 Montreal metropolitan hospitals. In both studies, a set of population-based controls randomly selected from the Quebec electoral list was established (Study 2: 762 cases and 899 controls); while in Study 1, an additional cancer control group was selected from a pool of other patients diagnosed with incident cancer (857 cases, 533 population controls, 1349 cancer controls). In both studies, detailed lifetime job histories were collected during interviews; a team of chemists and industrial hygienists then evaluated occupational exposure to many occupational substances including cleaning agents, biocides, aliphatic alcohols, ammonia, caustic soda, and waxes and polishes. Unconditional multivariate logistic regression was used to estimate odds ratios and 95% confidence intervals for lung cancer risk associated with various metrics of occupational exposure to these six agents, while adjusting for established risk factors. Interactions by smoking intensity and asthma status were explored with the inclusion of cross-product terms. Results: Overall, there was no consistent association supporting a role of occupational exposure to cleaning agents, biocides, and other cleaning-related agents in lung cancer etiology. Though there was some evidence that smoking intensity may modify the association between caustic soda and lung cancer risk in Study 1, where, among never-low intensity smokers, a threefold increase in risk was observed in comparison to a null association observed among medium-heavy intensity smokers (pInteraction=0.03). While, in Study 2, those occupationally exposed to aliphatic alcohols and who have ever had asthma experienced a four-fold increase in lung cancer risk in comparison to a null association observed among those who have never had asthma (pInteraction = 0.04). Conclusion: Taken together, the results of this study do not support the role of occupational exposure to cleaning agents, biocides, and other cleaning-related agents in lung cancer etiology.

Le risque de contamination des réseaux par les légionelles est aujourd'hui bien connu. Ces bactéries sont naturellement résistantes à un certain nombre de stress, elles peuvent survivre de façon prolongée dans les biofilms et proliférer de façon très importante via une croissance intracellulaire dans les amibes. Les amibes peuvent elles-mêmes présenter une résistance élevée aux traitements de décontamination usuels et offrent donc une protection supplémentaire aux légionelles. Si l'association amibes-légionelles dans les réseaux a été largement étudiée, il n'en est pas de même pour un certain nombre d'autres associations existant entre les amibes et des micro-organismes pathogènes avérés ou pathogènes émergents. Parmi les agents pathogènes émergents associés aux amibes, de nouvelles espèces bactériennes apparentées aux Chlamydia pourraient être responsables de pneumonies et d'avortements à répétition chez l'Homme et l'animal. Ces nouvelles espèces de Chlamydia présentent de nombreuses particularités, dont une survie à l'état libre dans l'environnement beaucoup plus importante que celles des Chlamydia " classiques " et une résistance importante aux températures élevées. De nouveaux virus " géants " associés aux amibes ont aussi été récemment décrits. Ils présentent de nombreuses particularités biologiques par rapport aux virus plus classiques et, bien que les preuves formelles de leur pathogénicité restent à apporter, certains de ces virus pourraient être responsables de pneumonies. Enfin, l'association amibes libres - mycobactéries soulève des questions importantes. Des travaux récents démontrent que des mycobactéries atypiques résistantes à certains traitements de décontamination (glutaraldéhyde notamment) sont aussi plus résistantes à certains traitements antibiotiques et plus à même de survivre dans les amibes. Par ailleurs la plupart des espèces de mycobactéries présentent la capacité de survivre dans les kystes amibiens, eux-mêmes très résistants à la plupart des traitements de décontamination. Les travaux de recherche présentés dans ce mémoire portent sur l'étude de l'écologie des amibes libres de l'environnement ainsi que sur l'étude de leur résistance à différents types de traitements biocides. Le rôle des amibes en tant que lieu d'échange de matériel génétique entre micro-organismes est abordé, ainsi que l'interaction des amibes avec les légionelles, les espèces apparentées aux Chlamydia, les virus d'amibes et les mycobactéries.

L'implication des tours aéro-réfrigérantes dites " ouvertes ", dans les cas de légionellose, demeure à l'heure actuel un problème de santé public malgré la multitude de composés biocides disponibles qui, parfois, sont écotoxiques et appliqués à de forte concentrations. L'efficacité de la majorité de ces biocides est étudiée vis-à-vis des formes planctoniques des microorganismes. Cette étude s'intéresse à la mise au point de deux procédés de traitement du risque Legionella pneumophila : H2O2/UV et ClO2, avec la prise en compte particulière des communautés microbiennes fixées et notamment des protozoaires qui constituent l'hôte naturel de cette bactérie. La démarche expérimentale adoptée pour l'étude de l'efficacité de ces traitements, s'est appuyée sur des biofilms environnementaux naturellement contaminés par des légionelles et issus d'une source thermale. La caractérisation des légionelles isolées des biofilms de cette source montre le maintien du sérogroupe 1 de L. pneumophila et l'apparition de deux autres sérogroupes non reportés dans des études précédentes (sg10 et sg12), avec toutefois une prédominance du sérogroupe 12. Quel que soit le sérogroupe, ces souches environnementales se sont avérées plus virulentes vis-à-vis de l'amibe Acantamoeba castellanii, que les souches cliniques de Lp1 répertoriées. A l'état planctonique elles se sont également avérées très sensibles aux traitements H2O2/UV et UV seul. A l'échelle expérimentale, les deux traitements, H2O2/UV et ClO2, montrent une performance élevée vis-à-vis des biofilms. L'étude a particulièrement mis en évidence le rôle du facteur trophique et de l'adaptation des bactéries au stress oxydatif dans la performance des traitements mais aussi dans l'apparition de la reviviscence. L'application des traitements à des installations en vraie grandeur a permis de conforter les résultats expérimentaux et de mettre l'accent, dans le contexte des sites étudiés, sur les limites de leur efficacité et sur la nécessité d'ajustements des doses appliquées (concentrations des biocides, flux d'irradiation UV, mode d'application) aux particularités des contextes industriels.

Στην παρούσα διατριβή διερευνήθηκαν οι παράμετροι που ελέγχουν τη δέσμευση/αποδέσμευση ιόντων Cu2+ ή/και κατιοντικών επιφανειοδραστικών ενώσεων, όπως το βρωμιούχο δεκαεξυλοτριμεθυλαμμώνιο (CTAB) ή το βρωμιούχο δεκαεξυλοτριμεθυλo φοσφώνιο (PC16) από αντίθετα φορτισμένους πολυηλεκτρολύτες. Mελετήθηκαν διαφορετικές πολυμερικές μήτρες που αποτελούνται είτε από εμπορικά διαθέσιμα πολυμερή είτε συντεθήκαν για αυτό το σκοπό, έτσι ώστε να γίνει κατανοητή η επίδραση της χημικής δομής του πολυμερούς στη ταχύτητα δέσμευσης και αποδέσμευσης των δραστικών συστατικών. Καθώς τα ιόντα αυτά έχουν βιοστατική δράση, απώτερος πρακτικός στόχος της διερεύνησης είναι η πιθανή εφαρμογή τέτοιων υβριδικών πολυμερικών υλικών σε υφαλοχώματα που θα εμποδίζουν την απόθεση θαλάσσιων βιοαποθέσεων και θα χαρακτηρίζονται από ιδιότητες «αυτοκαθαρισμού». Έτσι, παρασκευάστηκαν στατιστικά ή συσταδικά αμφίφιλα συμπολυμερή, στα οποία το υδρόφιλο μονομερές μπορεί να δεσμεύσει τα ιόντα Cu2+ ή/και τις αντίθετα φορτισμένες επιφανειοδραστικές ενώσεις, όπως το στυρενοσουλφονικό νάτριο (SSNa), το μηλεϊνικό (ΜΑc) και το ακρυλικό οξύ (ΑΑ), ενώ το υδρόφοβο μονομερές, το οποίο εισάγεται για τον έλεγχο του ισοζυγίου υδροφιλικότητας/υδροφοβικότητας, είναι ο μεθακρυλικός μεθυλεστέρας (ΜΜΑ). Έτσι, χρησιμοποιήθηκαν ομοπολυμερές PSSNa αλλά και συμπολυμερή του MAc με το SSNa (P(SSNa50-co-MANa). Επιπλέον, για συγκριτικούς λόγους παρασκευάστηκε και το συμπολυμερές του ΜΑc με οξικό βινυλεστέρα, (P(VAc-co-MAc), ενώ χρησιμοποιήθηκε και το εναλλασσόμενο συμπολυμερές του μηλεϊνικού οξέος με αιθυλένιο, P(Eth-alt-MAc). Τα στατιστικά αμφίφιλα συμπολυμερή P(MMAx-co-SSNa), P(MMAx-co-AA) παρασκευάστηκαν με συμπολυμερισμό ελευθέρων ριζών, ενώ τα συσταδικά αμφίφιλα συμπολυμερή PMMAx-b-PSSNa και PSSNa-b-PMMAx παρασκευάστηκαν με πολυμερισμό ελευθέρων ριζών μέσω μεταφοράς ατόμου (ΑΤRP), χρησιμοποιώντας ως μακροεκκινητή είτε PMMA είτε PSSNa, αντίστοιχα. Τα μοριακά βάρη των συμπολυμερών χαρακτηρίστηκαν με SEC, ενώ η χημική τους δομή ταυτοποιήθηκε με 1H-NMR και FT-IR. Η εκατοστιαία σύσταση των συμπολυμερών σε μονάδες ΜΜΑ, x, κυμαίνεται στην περιοχή 10-65 mol%, όπως προσδιορίστηκε με 1H-NMR και TGA. Από τη συγκριτική μελέτη της αυτοοργάνωσης σε υδατικό διάλυμα των συσταδικών και των στατιστικών αμφίφιλων συμπολυμερών, P(MMAx-co-SSNa) και PSSNa-b-PMMAx) με ιχνηθέτηση με φθορίζοντες ιχνηθέτες διαπιστώθηκε πως τα συσταδικά συμπολυμερή εμφανίζουν σημαντικότερη τάση αυτοοργάνωσης, καθώς έχουν σημαντικά μικρότερη κρίσιμη συγκέντρωση μικκυλοποίησης και χαρακτηρίζονται από υψηλότερα επίπεδα υδροφοβικότητας. Η δέσμευση των ιόντων Cu2+ από τα παραπάνω συμπολυμερή σε αραιά υδατικά διαλύματα πραγματοποιήθηκε μέσω ιονoανταλλαγής χρησιμοποιώντας τη μετά νατρίου άλατος μορφή των συμπολυμερών ή μέσω εξουδετέρωσης της όξινης μορφής των συμπολυμερών με Cu(OH)2 και διερευνήθηκε με φυσικοχημικές μεθόδους, όπως η θολομετρία, η ιξωδομετρία τριχοειδούς και η φασματοσκοπία υπεριώδους-ορατού. Διαπιστώθηκε πως σε αραιά διαλύματα η δέσμευση των ιόντων Cu2+ οδηγεί σε συρρίκνωση των πολυμερικών αλυσίδων (ιξωδομετρία τριχοειδούς). Μάλιστα, σε αντίθεση με τα συμπολυμερή που περιέχουν SSNa, στα πολυμερή που περιέχουν καρβοξυλικές ομάδες, η συρρίκνωση είναι ιδιαίτερα ισχυρή και οδηγεί σε διαχωρισμό φάσεων καθώς η αναλογία των ιόντων Cu2+ προς τις καρβοξυλικές ομάδες προσεγγίζει τη στοιχειομετρία (θολομετρία), ως αποτέλεσμα του σχηματισμού συμπλόκων ένταξης μεταξύ των ιόντων Cu2+ και των καρβοξυλικών ομάδων (φασματοσκοπία υπεριώδους-ορατού). Μάλιστα, στην περίπτωση του P(Eth-alt-MAc), η φασματοσκοπία υπεριώδους-ορατού έδωσε σαφείς ενδείξεις για το σχηματισμό διπύρηνων συμπλόκων χαλκού με τις ομάδες καρβοξυλίου, τα οποία έχουν τη δυνατότητα να δράσουν ως σταυροδεσμοί δικτυώνοντας τις πολυμερικές αλυσίδες. Ως αποτέλεσμα, σε ημιαραιά διαλύματα σχηματίζονται υδροπηκτώματα. Ιδιαίτερο ενδιαφέρον έχει το γεγονός πως οι ιξωδοελαστικές ιδιότητες των σχηματιζόμενων υδροπηκωτμάτων εξαρτώνται σημαντικά από το χρόνο ανάμιξης των συμπολυμερών με τα ιόντα Cu2+. Στην επόμενη φάση διερευνήθηκε ο σχηματισμός μικτών συσσωματωμάτων (συμπλόκων) μεταξύ των πολυηλεκτρολυτών που περιέχουν SSNa και της αντίθετα φορτισμένης επιφανειοδραστικής ένωσης, CTAΒ. Στην περίπτωση του ομοπολυμερούς PSSNa ή των συσταδικών αμφίφιλων συμπολυμερών PSSNa-b-PMMAx παρατηρείται διαχωρισμός φάσεων, καθώς η αναλογία των αλληλεπιδρώντων ειδών προσεγγίζει τη στοιχειομετρία φορτίων. Αντίθετα, όταν χρησιμοποιούνται τα στατιστικά αμφίφιλα συμπολυμερή P(MMAx-co-SSNa) με x=33-47mol%, σχηματίζονται ιξώδη διαλύματα ή ασθενή πηκτώματα. Η ρεολογική διερεύνηση του φαινόμενου κατέδειξε πως εξαρτάται από τη συγκέντρωση του πολυμερούς, τη γραμμομοριακή αναλογία CTAΒ/SSNa και την ιοντική ισχύ του υδατικού διαλύματος. Πριν τις τελικές δοκιμές σε πραγματικές συνθήκες παρασκευάστηκαν αιδιάλυτα στο νερό υβριδικά υλικά πολυμερούς-χαλκού, ενδεχομένως παρουσία CTAΒ ή PC16 χρησιμοποιώντας όλες τις προηγούμενες οικογένειες συμπολυμερών αλλά και εμπορικά διαθέσιμα πολυμερή αντίστοιχης δομής. Χρησιμοποιώντας κατάλληλο εκλεκτικό αντιδραστήριο (bathocuproine) μελετήθηκε φωτομετρικά η αποδέσμευση των ιόντων Cu2+ σε υδατικό διάλυμα NaCl 1 M. Έτσι, διευκρινίστηκε η επίδραση διαφόρων παραγόντων στο ρυθμό αποδέσμευσης των ιόντων Cu2+, όπως η ύπαρξη υδρόφοβων μονομερών, το είδος και το ποσοστό αυτών, η αρχιτεκτονική (στατιστική ή συσταδική) των συμπολυμερών, η εισαγωγή και το είδος της επιφανειοδραστικής ένωσης, ο τρόπος εισαγωγής και το είδος της αλληλεπίδρασης με τα ιόντα Cu2+. Στο τελευταίο στάδιο, διερευνήθηκε η συμβατότητά/αναμιξιμότητα των παρασκευασθέντων βιοστατικών υλικών με τυπικές πολυμερικές μήτρες που μπορούν να χρησιμοποιηθούν σε υφαλοχρώματα. Στις περιπτώσεις που διαπιστώθηκε ικανοποιητική συμβατότητα/αναμιξιμότητα (συστήματα PSSNa/CTAB, PSSNa/PC16, P(SSNa50-co-MANa)/CuPC16, P(VAc-co-MANa)/Cu) και ήταν εφικτό, παρασκευάστηκαν τα αντίστοιχα υφαλοχρώματα. Η συμπεριφορά αυτών των υφαλοχρωμάτων διερευνήθηκε εργαστηριακά με ηλεκτρονική μικροσκοπία σάρωσης (SEM), όσον αφορά τις μορφολογικές μεταβολές της επιφάνειας και τις μεταβολές του πάχους του χρώματος σε επιταχυνόμενες συνθήκες αποδέσμευσης βιοστατικού (υδατικό διάλυμα 5M NaCl). Επίσης, μελετάται και η συμπεριφορά τους σε πραγματικές συνθήκες με τη βοήθεια ειδικής διάταξης σε ελεγχόμενο θαλάσσιο χώρο. / At this thesis, the parameters that influence the interaction and the release of copper ions and/or of cationic surfactants as hexadecyltrimethyl ammonium bromide (CTAB) or hexadecyltrimethyl phosphonium bromide (PC16) from opposite charged polyelectrolytes, are studied. Different polymeric matrixes that are consisted either by commercially available polymers or by polymers that were synthesized for this purpose, were used in order, the influence of the polymer structure to the release rate of copper ions, to be identified. As these ions can behave as biocides, the practical purpose of this study is the possible application of these polymers to self-polishing antifouling paints. Random or block amphiphilic copolymers, whose hydrophilic monomer can interact with copper ions and/or the surfactants, such as styrene sulfonate (SSNa), maleic acid (MAc) and acrylic acid (AA), while the hydrophobic one, which is introduced for the hydrophilicity/hydrophobicity control, such as methyl methacrylate (MMA), were prepared. In addition, for comparison, the copolymer of maleic acid with vinyl acetate (VAc), (P(VAc-co-MAc)) was prepared. The alternative copolymer of maleic acid with ethylene was also used. The random copolymers P(MMAx-co-SSNa), P(MMAx-co-AA) were synthesized by free radical polymerization while the block copolymers PMMAx-b-PSSNa and PSSNa-b-PMMAx were synthesized through atom transfer polymerization (ATRP), using either PMMA or PSSNa as macroinitiator. The molecular weights of the copolymers were determined by SEC while their structure was identified by 1H-NMR and FT-IR. The copolymer composition in MMA untis is between 10-65 mol% as it was determined by 1H-NMR and TGA. The ability of self-assembling of random and block copolymers (P(MMAx-co-SSNa) και PSSNa-b-PMMAx) in aqueous solution was studied by pyrene fluorescence probing. The block copolymers was found to present self-assembling properties as they have lower critical micellization concentration. The binding of Cu2+ by the above mentioned polymers in dilute solutions was achieved through ion-exchange procedure using the sodium salt form of the copolymers or through neutralization of the acid form of the polymers using Cu(OH)2. Tholometry, viscometry and UV-Vis spectrometry were used to study this binding. The interaction of Cu2+ with the polymers leads to the shrinkage of the polymer chains. To the copolymers that contain carboxylic groups this shrinkage is more intense and leads to macroscopic phase separation, as the ratio of Cu2+ to the carboxylic groups is near the stoichiometry, as a consequence of complexes formation. The UV-Vis study of P(Eth-alt- MAc) denoted the formation of binuclear complexes between the Cu2+ and the carboxylic groups. These complexes can behave as crosslinks of the polymer chains. As a result, in semi-dilute solutions, gels are formed. Interestingly, the viscoelastic properties of these gels are dependent on the time elapsed from the mixture of the Cu2+ with the polymer. The interaction of polyelectrolytes that contain SSNa units, with opposite charged surfactant CTAΒ was studied. The interaction of homopolymer PSSNa or of the blocks copolymers PSSNa-b-PMMAx with CTAB leads to phase separation as the ratio of these compounds is near the stoichiometry. On the contrast, the interaction of CTAB with random copolymers P(MMAx-co- SSNa) where x=33-47mol%, leads to the formation of very viscous solutions or weak gels. By the study of the rheological properties of these systems, the influence of the polymer concentration, the molar ratio CTAΒ/SSNa and the ionic strength was found. All the above mentioned polymers where used for the preparation of hybrid materials that contain Cu2+. CTAΒ ή PC16 were also introduced to some of them in order to take a water insoluble material. The final purpose of this thesis was to control the release of Cu+2 ions from the prepared hybrid materials in aqueous NaCl 1M solution. The Cu+2 -release studies were performed photometrically using a selective dye (bathocuproine). As a result, the influence of many parameters such as the presence of hydrophobic monomers, their type and their content, the architecture of the copolymers (random or blocks), the introduction and the type of surfactants, the way of introduction of copper ions to the polymers and the type of their interaction was studied. The compatibility of all these materials with matrixes which are used in the antifouling paints was studied. In case that this compatibility was satisfying, antifouling paints using these polymeric materials (PSSNa/CTAB, PSSNa/PC16, P(SSNa50-co-MANa)/CuPC16, P(VAc-co-MANa)/Cu) were prepared. The morphology of the surface of these paints and the changes to their thickness was studied by SEM after staying in aqueous solution of NaCl 5M. Their behavior to real conditions (sea) is being studied.

Submitted by Priscila Nascimento (pnascimento@icict.fiocruz.br) on 2012-11-30T12:31:09Z No. of bitstreams: 1 beatriz-cyranka.pdf: 1720638 bytes, checksum: c436adb348584d5a5a8213fccdef6ea6 (MD5) / Made available in DSpace on 2012-11-30T12:31:09Z (GMT). No. of bitstreams: 1 beatriz-cyranka.pdf: 1720638 bytes, checksum: c436adb348584d5a5a8213fccdef6ea6 (MD5) Previous issue date: 2011 / Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil. / Novas tecnologias, como o sistema isolador para ensaios de esterilidade vieram a contribuir com a incorporação de um melhor desempenho destes ensaios no controle de qualidade das indústrias farmacêuticas.Este trabalho teve como objetivo a verificação do processo de biodescontaminação dentro do sistema isolador de Bio-Manguinhos, utilizado como agente esterilizante o gás de peróxido de hidrogênio. Foram utilizadas como biocarga para redução de contaminação microbiológica três concentrações dos microrganismos, Candida albicansATCC 10231, Clostridium sporogenesATCC 11437 eMicrococcus luteusATCC 9341, impregnadas em discos de filtro de celulose. Os estudos de cinética de crescimento dos microrganismos foram realizados para um melhor entendimento do seumetabolismo, bem como aspectos gerais de crescimento que contribuíram para ressaltar que a Candida albicans inicia sua fase exponencial de crescimento na segunda hora do cultivo e finaliza esta etapa na sexta hora do cultivo, com produção máxima de células viáveis, fato observado também no microrganismo Micrococccus luteus. Para o cultivo doClostridium sporogenes o crescimento foi mais lento com uma curva de crescimento com 60 horas de cultivo total. A produção de maior número de células para o Clostridiumfoi alcançada na vigésima quarta hora de cultivo, assim como a maior produção de esporos. Foi estabelecida, ao longo da curva de crescimento, a relação entre densidade ótica e número de células viáveis, relação essa importante para o estabelecimento das condições do estudo em relação à biocarga empregada de cada microrganismo no momento do desafio no sistema isolador. A capacidade de descontaminação avaliada dentro do sistema isolador com o biocida peróxido de hidrogênio revelou o tempo de exposição ao gás de 10 minutos, como resultado final satisfatório apresentando redução total da carga microbiana com destruição total das células viáveis, assim como as formas esporuladas do Clostridium sporogenes. Desta forma conclui-se que o peróxido de hidrogênioé um biocida de eficácia comprovada, nas variáveis deste estudo e o processode descontaminação no sistema isolador de Bio-Manguinhos é compatível comsua atividade finalística na produção de insumos para a saúde. / New technologies such as isolator system for sterility tests came to help with the incorporation of a better performance of these kinds of tests in quality control of pharmaceutical companies. This study aimed to verify the decontamination process within the system isolator of Bio-Manguinhos, usinghydrogen peroxide gas as a sterilizing agent. Three concentrations of microorganisms were used as bioburden for the microbiological contamination reduction, Candida albicansATCC 10231, Clostridium sporogenesATCC 11437 and Micrococcus luteusATCC 9341, impregnated in cellulose filter discs. Studies of growth kinetics of microorganisms were carried out to a better understanding of theirmetabolism, as well as general aspects of growth that contributed to emphasize that the Candida albicansbegins its exponential growth phase in the second hour of cultivation and this step ends at the sixth hour cultivation, with maximum yield of viable cells, this was also observed in the Micrococcus luteusmicroorganism. For the Clostridium sporogenescultivation, growth was slower with a 60 hours growth curve of total culture. The production of more cells for Clostridiumwas achieved in the twenty-fourth hour of cultivation, as well as the maximum spores production. It was established along the growth curve the relationship between optical density and numberof viable cells, this relationship was important to establish the conditions of the study related to the bioburden of each microorganism used to challenge the isolator system. The decontamination capacity evaluated within the isolator system with the hydrogen peroxide biocide showed that the gas exposure time of 10 minutes wassatisfactory demonstrating total reduction of the microbial load with total destruction of viable cells, as well as the sporulated forms of Clostridium sporogenes. Thus it is concluded that hydrogen peroxide is a proved effective biocide, in the variables of this study and decontamination process in the Bio-Manguinhos insulator system is compatible with its main activity in the production of health supplies.