The aim of this diploma thesis is to optimize a method of interfacial rheology for testing the interfacial behaviour of biofilms on the liquid-air interface and after that use the method for studying the biofilm formation under optimal and stress conditions. For studying the biofilm formation, Kombucha was used. It is a microbial culture forming a cellulose biofilm on the interface. As the stress conditions, reduction of sucrose concentration, change of pH and change of ionic strength was used. Next, the ability of regeneration of biofilm formed on the interface was studied. The biofilm formation was occured in all cases except of increasing ionic strength. As expected, the best biofilm biofilm growth was observed under optimal condition, which means a sucrose concentration 100 g/l.

As infecções causadas por leveduras representam um grave problema em Saúde Pública. O objetivo desta pesquisa é o estudo aderência de Candida spp. (C. albicans ATCC18804, C. albicans SC5314, C. albicans cepa de campo, C. glabrata ATCC2001) sobre cateter urinário de Látex siliconizado (LS) e Silicone total (ST); avaliação da sensibilidade ao M-EDTA® dos biofilmes formados e quantificação dos genes ALS1 e ALS3 expressos por células sésseis de C. albicans. Os biofilmes de Candida spp. foram produzidos sobre cateteres urinários (LS e ST) segmentados em 1,0 cm de comprimento e separadamente introduzidos em poços de placas de poliestireno, sendo incubados a 35±1°C em contato com as suspensões de leveduras por 6, 24 e 48 e 72 horas. Os segmentos de cateter foram examinados por MEV, por cultura microbiológica, para avaliação da viabilidade celular e redução do sal de tetrazólio (XTT), para medir a atividade metabólica das células em biofilme. A sensibilidade das células sésseis ao M-EDTA® foi determinada após a exposição dos biofilmes ao produto; a expressão dos genes ALS1 e ALS3 de C. albicans SC5314 foi analisada por reação em cadeia de polimerase quantitativo (qPCR) e o resultado da recuperação de células viáveis de biofilme de Candida spp. foi expresso em log média ± desvio-padrão. Do biofilme de C. albicans ATCC18804 formado sobre LS, a recuperação foi de 4,48±2,52 e sobre ST; 4,76±1,62. A média de recuperação do biofilme de C. albicans SC5314 sobre LS foi de 4,35±0,39 e sobre ST, de 4,21±0,22. Para C. albicans cepa de campo, a média no LS foi 5,15±0,18 e no ST; 4,79±0,17. Do biofilme de C. glabrata ATCC2001 formado sobre o LS, a recuperação média foi de 4,69±0,19 e sobre ST, de 4,75±0,68. A atividade metabólica das células em biofilme foi expressa em valores de absorbância. Células de C. albicans ATCC18804 aderidas ao LS apresentaram atividade de 0,070±0,06 e sobre ST; 0,0681±0,06. A atividade metabólica do biofilme de C. albicans SC5314 sobre o LS foi de 0,0622±0,05 e sobre ST; 0,0785±0,05. Das células de C. albicans cepa de campo aderidas ao LS detectou-se atividade de 0,1064±0,01 e sobre ST, de 0,904±0,03. Células de C. glabrata ATCC2001 aderidas ao LS apresentaram uma atividade de 0,0785±0,04 e sobre ST; 0,0755±0,03. A observação ao MEV mostrou células leveduriformes de C. albicans SC5314 aderidas ao LS e ao ST formando monocamadas. Candida albicans ATCC18804, C. albicans cepa de campo e C. glabrata ATCC2001 formaram monocamadas de células leveduriformes e produziram filamentos sobre a superfície LS e ST, sugerindo uma estrutura tridimensional do biofilme. A exposição ao M-EDTA® dos biofilmes de C. albicans ATCC18804, C. albicans SC5314, C. albicans cepa de campo e C. glabrata ATCC2001 mostrou que os mesmo foram sensíveis à ação do produto. Após o crescimento de C. albicans SC5314 em biofilme, não houve detecção de expressão dos genes ALS1 e ALS3. Em conclusão, todas as espécies de Candida formaram biofilme sobre os biomateriais. Os biofilmes foram sensíveis a ação do M-EDTA® e C. albicans SC5314 não expressou os genes ALS1 e ALS3. / Infections caused by yeasts represent a serious problem in Public Health. The aim of this research was to study the Candida spp. (C. albicans ATCC18804, C. albicans SC5314, C. albicans wild type, C. glabrata ATCC2001) attachment onto urinary catheters made of Siliconized latex (LS) and Total silicon (ST); evaluating the susceptibility of biofilms to M-EDTA® and quantifying genes ALS1 and AL3 expressed by sessile cells of C. albicans. The Candida spp. biofilms were produced over the surface of urinary catheters (LS and ST). The catheters were cut in 1.0cm in length, introduced in wells of polystyrene plates and incubated at 35±1°C in contact with yeasts suspensions during 6, 24, 48 and 72 hours. The catheter samples were examined by SEM, by microbiology culture to evaluate the viable cells and reduction of tetrazolium salt (XTT) to measure the metabolic activity of Candida spp. cells into biofilms. The sensitivity of sessile cells to M-EDTA® was determinated after the exposition of biofilms to the product. The C. albicans SC5314 expression of ALS1 and ALS3 genes was analyzed by quantitative polymerase chain reaction (qPCR). The results of Candida spp. viable cells recovery were determined in log average ± standard deviation. From C. albicans ATCC18804 biofilm cells formed onto LS, the recovery was 4.48±2.52 e onto ST; 4.76±1.62. The average of recovery from C. albicans SC5314 biofilm formed onto LS was 4.35±0.39 and onto ST; 4.21±0.22. For C. albicans wild type, the recovery onto LS was 5.15±0.18 and above ST; 4.79±0.17. C. glabrata ATCC2001 biofilm formed onto LS, the medium was 4.69±0.19 and onto ST; 4.75±0.68. The metabolic activity from cells into biofilms was determined in absorbance values. C. albicans ATCC18804 cells attachment on LS had an activity of 0.070±0.06 and on ST; 0.0681±0.06. The metabolic activity of C. albicans SC5314 biofilm formed onto LS was 0.0622±0.05 and onto ST; 0.0785±0.05. From C. albicans wild type attached onto LS, the activity detected was 0.1064±0.01 and on ST; 0.904±0.03. C. glabrata ATCC2001 cells attached on LS had an activity of 0.0785±0.04 and on ST; 0.0755±0.03. The SEM observations showed C. albicans SC5314 yeast cells attached on LS and ST forming a monolayer. C. albicans ATCC18804, C. albicans wild type and C. glabrata ATCC2001 formed yeast monolayer and produced filaments over the surface of LS and ST, suggesting the three-dimensional structure of biofilm. The M-EDTA® exposition of C. albicans ATCC18804, C. albicans SC5314, C. albicans wild type and C. glabrata ATCC2001 showed that the biofilms were sensitive to the product action. After the C. albicans SC5314 biofilm growth, there was not expression of the ALS1 and ALS3 genes. In conclusion, all the Candida species formed biofilms on the biomaterials. The biofilms were sensitive to M-EDTA® and the C. albicans SC5314 didn´t express the ALS1 and ALS3 genes.

As infecções causadas por leveduras representam um grave problema em Saúde Pública. O objetivo desta pesquisa é o estudo aderência de Candida spp. (C. albicans ATCC18804, C. albicans SC5314, C. albicans cepa de campo, C. glabrata ATCC2001) sobre cateter urinário de Látex siliconizado (LS) e Silicone total (ST); avaliação da sensibilidade ao M-EDTA® dos biofilmes formados e quantificação dos genes ALS1 e ALS3 expressos por células sésseis de C. albicans. Os biofilmes de Candida spp. foram produzidos sobre cateteres urinários (LS e ST) segmentados em 1,0 cm de comprimento e separadamente introduzidos em poços de placas de poliestireno, sendo incubados a 35±1°C em contato com as suspensões de leveduras por 6, 24 e 48 e 72 horas. Os segmentos de cateter foram examinados por MEV, por cultura microbiológica, para avaliação da viabilidade celular e redução do sal de tetrazólio (XTT), para medir a atividade metabólica das células em biofilme. A sensibilidade das células sésseis ao M-EDTA® foi determinada após a exposição dos biofilmes ao produto; a expressão dos genes ALS1 e ALS3 de C. albicans SC5314 foi analisada por reação em cadeia de polimerase quantitativo (qPCR) e o resultado da recuperação de células viáveis de biofilme de Candida spp. foi expresso em log média ± desvio-padrão. Do biofilme de C. albicans ATCC18804 formado sobre LS, a recuperação foi de 4,48±2,52 e sobre ST; 4,76±1,62. A média de recuperação do biofilme de C. albicans SC5314 sobre LS foi de 4,35±0,39 e sobre ST, de 4,21±0,22. Para C. albicans cepa de campo, a média no LS foi 5,15±0,18 e no ST; 4,79±0,17. Do biofilme de C. glabrata ATCC2001 formado sobre o LS, a recuperação média foi de 4,69±0,19 e sobre ST, de 4,75±0,68. A atividade metabólica das células em biofilme foi expressa em valores de absorbância. Células de C. albicans ATCC18804 aderidas ao LS apresentaram atividade de 0,070±0,06 e sobre ST; 0,0681±0,06. A atividade metabólica do biofilme de C. albicans SC5314 sobre o LS foi de 0,0622±0,05 e sobre ST; 0,0785±0,05. Das células de C. albicans cepa de campo aderidas ao LS detectou-se atividade de 0,1064±0,01 e sobre ST, de 0,904±0,03. Células de C. glabrata ATCC2001 aderidas ao LS apresentaram uma atividade de 0,0785±0,04 e sobre ST; 0,0755±0,03. A observação ao MEV mostrou células leveduriformes de C. albicans SC5314 aderidas ao LS e ao ST formando monocamadas. Candida albicans ATCC18804, C. albicans cepa de campo e C. glabrata ATCC2001 formaram monocamadas de células leveduriformes e produziram filamentos sobre a superfície LS e ST, sugerindo uma estrutura tridimensional do biofilme. A exposição ao M-EDTA® dos biofilmes de C. albicans ATCC18804, C. albicans SC5314, C. albicans cepa de campo e C. glabrata ATCC2001 mostrou que os mesmo foram sensíveis à ação do produto. Após o crescimento de C. albicans SC5314 em biofilme, não houve detecção de expressão dos genes ALS1 e ALS3. Em conclusão, todas as espécies de Candida formaram biofilme sobre os biomateriais. Os biofilmes foram sensíveis a ação do M-EDTA® e C. albicans SC5314 não expressou os genes ALS1 e ALS3. / Infections caused by yeasts represent a serious problem in Public Health. The aim of this research was to study the Candida spp. (C. albicans ATCC18804, C. albicans SC5314, C. albicans wild type, C. glabrata ATCC2001) attachment onto urinary catheters made of Siliconized latex (LS) and Total silicon (ST); evaluating the susceptibility of biofilms to M-EDTA® and quantifying genes ALS1 and AL3 expressed by sessile cells of C. albicans. The Candida spp. biofilms were produced over the surface of urinary catheters (LS and ST). The catheters were cut in 1.0cm in length, introduced in wells of polystyrene plates and incubated at 35±1°C in contact with yeasts suspensions during 6, 24, 48 and 72 hours. The catheter samples were examined by SEM, by microbiology culture to evaluate the viable cells and reduction of tetrazolium salt (XTT) to measure the metabolic activity of Candida spp. cells into biofilms. The sensitivity of sessile cells to M-EDTA® was determinated after the exposition of biofilms to the product. The C. albicans SC5314 expression of ALS1 and ALS3 genes was analyzed by quantitative polymerase chain reaction (qPCR). The results of Candida spp. viable cells recovery were determined in log average ± standard deviation. From C. albicans ATCC18804 biofilm cells formed onto LS, the recovery was 4.48±2.52 e onto ST; 4.76±1.62. The average of recovery from C. albicans SC5314 biofilm formed onto LS was 4.35±0.39 and onto ST; 4.21±0.22. For C. albicans wild type, the recovery onto LS was 5.15±0.18 and above ST; 4.79±0.17. C. glabrata ATCC2001 biofilm formed onto LS, the medium was 4.69±0.19 and onto ST; 4.75±0.68. The metabolic activity from cells into biofilms was determined in absorbance values. C. albicans ATCC18804 cells attachment on LS had an activity of 0.070±0.06 and on ST; 0.0681±0.06. The metabolic activity of C. albicans SC5314 biofilm formed onto LS was 0.0622±0.05 and onto ST; 0.0785±0.05. From C. albicans wild type attached onto LS, the activity detected was 0.1064±0.01 and on ST; 0.904±0.03. C. glabrata ATCC2001 cells attached on LS had an activity of 0.0785±0.04 and on ST; 0.0755±0.03. The SEM observations showed C. albicans SC5314 yeast cells attached on LS and ST forming a monolayer. C. albicans ATCC18804, C. albicans wild type and C. glabrata ATCC2001 formed yeast monolayer and produced filaments over the surface of LS and ST, suggesting the three-dimensional structure of biofilm. The M-EDTA® exposition of C. albicans ATCC18804, C. albicans SC5314, C. albicans wild type and C. glabrata ATCC2001 showed that the biofilms were sensitive to the product action. After the C. albicans SC5314 biofilm growth, there was not expression of the ALS1 and ALS3 genes. In conclusion, all the Candida species formed biofilms on the biomaterials. The biofilms were sensitive to M-EDTA® and the C. albicans SC5314 didn´t express the ALS1 and ALS3 genes.

Les réseaux trophiques jouent un rôle primordial dans la régulation des flux de matière et d’énergie au sein des écosystèmes. Dans le cadre des pratiques de recharge artificielle des aquifères, les biocénoses souterraines sont pleinement sollicitées et leur capacité à dégrader les flux de matière organique de surface conditionne le maintien de la qualité des eaux souterraines. L’objectif de ce travail est de déterminer l’influence d’une augmentation des flux de carbone organique dissous sur l’intensité des interactions trophiques entre les communautés microbiennes et les assemblages d’invertébrés au toit des nappes phréatiques rechargées artificiellement avec des eaux de ruissellement pluvial. A travers une approche expérimentale de terrain et de laboratoire, ce travail permet d’évaluer l’intensité des relations existant entre les flux de carbone organique dissous, les conditions environnementales, l’activité et la diversité de communautés microbiennes et l’abondance des communautés d’invertébrés. / Food webs play a crucial role in regulating the fluxes of matter and energy within ecosystems. Artificial recharge of aquifers relies heavily on the ability of groundwater biocenoses to degrade organic matter fluxes that is a condition to maintain the quality of groundwater. The objective of this work is to determine the impact of increased dissolved organic carbon supply on the trophic interactions between the microbial communities and invertebrate assemblages at the upper layers of groundwater artificially recharged with stormwater. Through a combined field and laboratory experimental approach, this work allows ranking the strength of relationship between dissolved organic carbon fluxes, environmental conditions, activity and diversity of microbial communities and abundance of invertebrate assemblages