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While sea spray aerosols (SSAs) generation by oceanic breaking waves continues to be an active research area, lake spray aerosols (LSAs) production by freshwater breaking waves is an emerging research field. Recent studies have linked LSAs to regional cloud processes and the aerosolization of freshwater pathogens and pollutants. Yet, differences in spray aerosol ejection between freshwater and saltwater and their impact on the water-to-air dispersal of microorganisms and pollutants are poorly understood. The goals of this dissertation work were to understand mechanistic differences between spray aerosol generation in freshwater and saltwater, develop a representation of LSA emissions in atmospheric models and evaluate their impact on regional aerosol loading, and compare the aerosolization of bacteria and microplastics by SSAs and LSAs. Experiments in a breaking-waves analogue tank revealed that the subsurface bubble plume in saltwater is characterized by more submillimeter bubbles than that in freshwater, and hence, saltwater surface foams were more persistent and were comprised of more submillimeter surface bubbles. Consequently, the average number concentration of generated SSAs was eight times higher than that of LSAs. Using these measurements, the developed LSA emission parametrization revealed that freshwater emissions were, at least, an order of magnitude lower than saltwater emissions for the same wave-breaking conditions. When implementing this emission parameterization to simulate LSA emissions from the Laurentian Great Lakes, LSAs did not contribute significantly to regional aerosol loading (< 2%), yet their impact on coarse-mode aerosols was more significant with up to a 19-fold increase in some areas. Furthermore, modeled LSAs reached up to 1000 km inland and were incorporated in the lakes' cloud layer. Despite the generation of more spray aerosols in saltier waters, cumulative salt additions in the freshwater–saltwater continuum (0-35 g/kg) led to a nonmonotonic increase in freshwater bacterial aerosolization abundance, which exhibited a peak at lower oligohaline conditions (0.5-1 g/kg). However, the aerosolization of microplastics by SSAs was one order of magnitude higher than that by LSAs. Overall, this dissertation work showed that LSA emissions are intrinsically different from SSA emissions, which influences their role in transferring microorganisms and pollutants at the air-water interface. / Doctor of Philosophy / When waves break, they entrain large volumes of air in the form of subsurface bubbles. These bubbles rise to the surface and pop ejecting small droplets into the air, also known as spray aerosols. The droplets ejected from saltwater breaking waves are referred to as sea spray aerosols (SSAs) and are extensively studied due to their important role in Earth's atmosphere. However, the ejection of lake spray aerosols (LSAs) from freshwater breaking waves is far less understood. With recent studies linking freshwater breaking waves to regional cloud processes and the transfer of aquatic pathogens to the air, a better understanding of LSAs formation and how it compares to SSAs production was needed. The goals of this dissertation work were to understand the differences between spray aerosol generation in freshwater and saltwater, develop a representation of LSA emissions in atmospheric models and assess their contribution to atmospheric aerosols, and contrast the role of LSAs and SSAs in transferring bacteria and microplastics to the air. Experiments in a spray aerosol generation tank revealed that saltwater breaking waves generate more submillimeter bubbles at the subsurface level than freshwater breaking waves and that the generated surface foam is more persistent and is comprised of smaller bubbles in saltwater. Consequently, SSA generation in the experimental tank was eight times higher than LSA generation. When implementing these results in an atmospheric model to simulate LSA emission from the surface of the Laurentian Great Lakes, it was found that the regional aerosol population was not significantly affected. However, LSA particles were transported inland up to 1000 km and reached cloud level which hints at possible implications on public health and regional climate. Despite a higher generation of aerosols by breaking waves in saltier waters, the abundance of freshwater bacteria that was dispersed to the air by spray aerosols did not increase monotonically in response to a gradual increase in freshwater salinity. Yet, microplastics transfer to the air by SSAs was an order of magnitude higher than that by LSAs. The results of this dissertation work highlight the important differences between the generation of spray aerosols by breaking waves in freshwater and saltwater and their corresponding roles in the water-to-air dispersal of microorganisms and pollutants.

La qualité de l’air dans les habitats est un problème majeur en Europe, où les personnes passent une grande partie de leur temps en milieu intérieur. Selon l’OMS, de nombreux habitats sont touchés par l’humidité entraînant le développement de moisissures qui ont des conséquences sanitaires et économiques.Dans ce travail, une approche globale, associant étude de terrain et expérimentations en laboratoire, a été employée pour décrire l’exposition aux bioaérosols dans des habitations dégradées par les moisissures, étudier leurs effets sur la santé des résidents et proposer une démarche diagnostique applicable aux habitats dégradés. Nous avons ainsi mis en évidence la présence récurrente de certaines espèces fongiques, notamment Aspergillus versicolor, Penicillium chrysogenum et P. crustosum, qui pourraient être utilisées comme indicateurs microbiologiques de contamination fongique de l’air intérieur. Par ailleurs les analyses statistiques ont permis de montrer des liens entre les concentrations de certaines espèces de micromycètes dans l’air et les catégories de surface contaminées proposées par l’ANSES. Des relations ont également pu être établies entre l’exposition à ces moisissures et certaines manifestations respiratoires et cutanées mentionnées chez les résidents. L’évaluation du potentiel cytotoxique des bioaérosols collectés dans les habitats dégradés a, quant à elle, permis de mettre en évidence un lien entre les réponses observées sur deux lignées cellulaires testées (pulmonaire A549 et cutanée HaCaT).D’un point de vue méthodologique, la qPCR couplée à la cytométrie en flux semble être une méthode rapide qui peut être corrélée à l’approche culturale et permet ainsi de suivre l’exposition à Aspergillus versicolor dans les habitats dégradés. Huit espèces fongiques récurrentes dans les bioaérosols ont également été sélectionnées pour une étude en enceinte climatique montrant l’impact de la température et de l’humidité relative sur la croissance et la toxinogenèse fongique. / Air quality in houses is a major concern in Europe, as people spend most of their time indoors. According to the WHO, numerous houses are exposed to dampness that can lead to mold growth, causing health and economic damages.In this work, a global approach including both field study and laboratory experimentations was used to characterize the human exposure to bioaerosols in mold-damaged houses, to study their health effects on inhabitants and to propose a diagnostic process that could be applied to mold-damaged houses. We showed the recurrence of several fungal species, specifically Aspergillus versicolor, Penicillium chrysogenum and P. crustosum, which could be used as microbial indicators of airborne fungal contamination. In addition, our statistical analyses showed relations between the concentrations of recurrent molds in air and the levels of surface contamination by molds proposed by ANSES. Statistical link has also been found between mold exposure and respiratory or cutaneous symptoms described by the inhabitants. A cytotoxic potential evaluation of bioaerosols collected in these mold-damaged houses allowed to highlight a relation between the responses observed for the two cell lines (pulmonary A549 and cutaneous HaCaT).From a methodological point of view, qPCR coupled with flow cytometry appears to be a fast and reliable method that can be correlated to cultural approach, allowing to monitor the human exposure to Aspergillus versicolor in mold-damaged buildings. Eight recurrent fungal species identified in bioaerosols were also selected for a study in a climatic chamber that showed the effects of temperature and relative humidity on fungal growth and toxinogenic potential.

Há poucos estudos relacionados com a caracterização dos componentes biológicos presentes no material particulado atmosférico, chamados bioaerossóis. Esses componentes biológicos podem assumir uma relação direta com a deflagração de doenças e também estão relacionados com os processos climáticos, em especial como núcleos de formação de gelo. Dentre os bioaerossóis, foram estudados, neste trabalho, os fungos anemófilos, sendo os principais os Basidiomycota, Ascomycota e os Fungos mitospóricos. O objetivo desse trabalho foi estimar a contribuição dos aerossóis fúngicos para a composição biogênica do material particulado atmosférico da Região Metropolitana de São Paulo. Foram realizadas medidas em dois sítios: na Cidade Universitária e no Pico do Jaguará. Para isso, foram caracterizados os tipos fúngicos utilizando-se um microscópio ótico e estimada a massa dos aerossóis de origem fúngica utilizando-se biomarcadores. Foram realizadas coletas durante 2013, 2014 e 2015 com o amostrador Burkard 7-day Recording Sampler, sendo que em 2015 também foram realizadas coletas com filtros de quartzo com o amostrador Airmetrics MiniVol portable Sampler. Foram identificados e obtida a concentração dos tipos fúngicos observados na atmosfera da RMSP, sendo encontrados 39 grupos principais de fungos, sendo os principais os Basidiomycotas. A concentração média total foi de 5736 (± 2459) esporos/m³ por dia. Os Ascomycota, Basidiomycota e os fungos mitospóricos se correlacionaram com as variáveis meteorológicas de forma diferente. Observou-se variação da concentração de esporos durante os diferentes períodos do dia, sendo que a maior concentração de esporos ocorreu na madrugada, possivelmente devido às condições meteorológicas (elevada umidade e temperaturas mais amenas), contudo a concentração dos fungos mitospóricos foi maior durante o período da tarde, principalmente devido aos mecanismos de liberação de esporos. No verão e na primavera foram obtidas as maiores concentrações de Ascósporos e Basidiósporos e no inverno e outono foram obtidas as maiores concentrações dos fungos mitospóricos. Além disso, observou-se uma correlação positiva e significativa entre a concentração de esporos totais e o material particulado inalável (MP10), indicando que ambos são influenciados da mesma forma pelas variáveis meteorológicas e/ou tem uma fonte em comum, como por exemplo a ressuspensão da poeira do solo. O Arabitol, Manitol, Treitol e os esporos de fungos são positivamente correlacionados, indicando que os três açucares podem ser traçadores de esporos de fungos na atmosfera. Utilizando-se um fator de conversão proposto na literatura foi possível estimar que 2% da concentração do MP10 e 8% da concentração do OC estão associados aos esporos de fungos, indicando a importância dos esporos fúngicos. Com o uso de modelos receptores foram observadas 6 fontes para o MP10: queima de biomassa resultante de processos industriais; queima de biomassa resultante da queima da vegetação; aerossóis fúngicos; formação de aerossol secundário; ressuspensão do solo e emissão veicular. Esses resultados constituem um avanço para a pesquisa em bioaerossóis no Brasil, já que muitas dessas análises ainda não haviam sido realizadas em áreas urbanas no Brasil / Although the biogenic components of Particulate Matter (PM) can have a direct relationship with the outbreak of respiratory diseases and can be linked to changes in climate processes there are very few studies related to its characterization in the atmospheric aerosols. Among the bioaerosols, the airborne fungi were studied, and the Basidiomycota, Ascomycota and Mitosporic fungi were the main characterized. The goal of this work was to estimate the contribution of the fungal aerosol to the RMSP atmosphere, with samplings in two sites: Cidade universitária (USP) and Pico do Jaraguá (PJ). For that, the fungi types were characterized using an optical microscope and it was estimated the mass of the fungal aerosol using biomarkers. Sampling was carried out during 2013, 2014 and 2015 with the \"Burkard 7-day Recording Sampler\", and in 2015, filters were sampled with the \"Airmetrics MiniVol portable Sampler. It was identified and determined the concentration of the fungi types observed in the RMSP atmosphere, and were found 39 main groups of fungi. The main group was the Basidiomycota. The mean concentration of the total fungi was 5736 (± 2459) spores/m³ per day. The Ascomycota, Basidiomycota and the mitosporic fungi correlated in different ways with the meteorological variables. A variation of the spores concentration during different times of the day was observed, with the occurrence of the highest concentration of spores at dawn, possibly due to weather conditions (high humidity and cooler temperatures), but the concentration of the mitosporic fungi was higher during the afternoon, mainly due to the spores release mechanisms. In the summer and spring were observed higher concentrations of Ascospores and Basidiospores and in the autumn and winter were obtained higher concentration of Mitospores. Besides that, it was also observed a positive and significant correlation between the fungal spore with the particulate matter, indicating that both are influenced in the same way by weather variables and/or has a common source such as soil resuspension. Arabitol, Mannitol, Threitol and the fungi spores are positive correlated, indicating that these three sugar alcohols can be a tracer for fungi spores in the atmosphere. Using the conversion factor proposed in the literature was possible to estimate that 2% of the PM10 concentration and 8% of the OC concentration it is due to fungi spores, indicating its importance. Using receptor modeling it was identified six sources for PM10, the biomass burning resulting from industrial processes, the biomass burning resulting from vegetation burning, the fungal aerosol, the secondary formation aerosol, the soil resuspension and vehicular emission. These results are a breakthrough for research on bioaerosols in Brazil, since many of this analyzes had never been performed in urban areas in Brazil

Há poucos estudos relacionados com a caracterização dos componentes biológicos presentes no material particulado atmosférico, chamados bioaerossóis. Esses componentes biológicos podem assumir uma relação direta com a deflagração de doenças e também estão relacionados com os processos climáticos, em especial como núcleos de formação de gelo. Dentre os bioaerossóis, foram estudados, neste trabalho, os fungos anemófilos, sendo os principais os Basidiomycota, Ascomycota e os Fungos mitospóricos. O objetivo desse trabalho foi estimar a contribuição dos aerossóis fúngicos para a composição biogênica do material particulado atmosférico da Região Metropolitana de São Paulo. Foram realizadas medidas em dois sítios: na Cidade Universitária e no Pico do Jaguará. Para isso, foram caracterizados os tipos fúngicos utilizando-se um microscópio ótico e estimada a massa dos aerossóis de origem fúngica utilizando-se biomarcadores. Foram realizadas coletas durante 2013, 2014 e 2015 com o amostrador Burkard 7-day Recording Sampler, sendo que em 2015 também foram realizadas coletas com filtros de quartzo com o amostrador Airmetrics MiniVol portable Sampler. Foram identificados e obtida a concentração dos tipos fúngicos observados na atmosfera da RMSP, sendo encontrados 39 grupos principais de fungos, sendo os principais os Basidiomycotas. A concentração média total foi de 5736 (± 2459) esporos/m³ por dia. Os Ascomycota, Basidiomycota e os fungos mitospóricos se correlacionaram com as variáveis meteorológicas de forma diferente. Observou-se variação da concentração de esporos durante os diferentes períodos do dia, sendo que a maior concentração de esporos ocorreu na madrugada, possivelmente devido às condições meteorológicas (elevada umidade e temperaturas mais amenas), contudo a concentração dos fungos mitospóricos foi maior durante o período da tarde, principalmente devido aos mecanismos de liberação de esporos. No verão e na primavera foram obtidas as maiores concentrações de Ascósporos e Basidiósporos e no inverno e outono foram obtidas as maiores concentrações dos fungos mitospóricos. Além disso, observou-se uma correlação positiva e significativa entre a concentração de esporos totais e o material particulado inalável (MP10), indicando que ambos são influenciados da mesma forma pelas variáveis meteorológicas e/ou tem uma fonte em comum, como por exemplo a ressuspensão da poeira do solo. O Arabitol, Manitol, Treitol e os esporos de fungos são positivamente correlacionados, indicando que os três açucares podem ser traçadores de esporos de fungos na atmosfera. Utilizando-se um fator de conversão proposto na literatura foi possível estimar que 2% da concentração do MP10 e 8% da concentração do OC estão associados aos esporos de fungos, indicando a importância dos esporos fúngicos. Com o uso de modelos receptores foram observadas 6 fontes para o MP10: queima de biomassa resultante de processos industriais; queima de biomassa resultante da queima da vegetação; aerossóis fúngicos; formação de aerossol secundário; ressuspensão do solo e emissão veicular. Esses resultados constituem um avanço para a pesquisa em bioaerossóis no Brasil, já que muitas dessas análises ainda não haviam sido realizadas em áreas urbanas no Brasil / Although the biogenic components of Particulate Matter (PM) can have a direct relationship with the outbreak of respiratory diseases and can be linked to changes in climate processes there are very few studies related to its characterization in the atmospheric aerosols. Among the bioaerosols, the airborne fungi were studied, and the Basidiomycota, Ascomycota and Mitosporic fungi were the main characterized. The goal of this work was to estimate the contribution of the fungal aerosol to the RMSP atmosphere, with samplings in two sites: Cidade universitária (USP) and Pico do Jaraguá (PJ). For that, the fungi types were characterized using an optical microscope and it was estimated the mass of the fungal aerosol using biomarkers. Sampling was carried out during 2013, 2014 and 2015 with the \"Burkard 7-day Recording Sampler\", and in 2015, filters were sampled with the \"Airmetrics MiniVol portable Sampler. It was identified and determined the concentration of the fungi types observed in the RMSP atmosphere, and were found 39 main groups of fungi. The main group was the Basidiomycota. The mean concentration of the total fungi was 5736 (± 2459) spores/m³ per day. The Ascomycota, Basidiomycota and the mitosporic fungi correlated in different ways with the meteorological variables. A variation of the spores concentration during different times of the day was observed, with the occurrence of the highest concentration of spores at dawn, possibly due to weather conditions (high humidity and cooler temperatures), but the concentration of the mitosporic fungi was higher during the afternoon, mainly due to the spores release mechanisms. In the summer and spring were observed higher concentrations of Ascospores and Basidiospores and in the autumn and winter were obtained higher concentration of Mitospores. Besides that, it was also observed a positive and significant correlation between the fungal spore with the particulate matter, indicating that both are influenced in the same way by weather variables and/or has a common source such as soil resuspension. Arabitol, Mannitol, Threitol and the fungi spores are positive correlated, indicating that these three sugar alcohols can be a tracer for fungi spores in the atmosphere. Using the conversion factor proposed in the literature was possible to estimate that 2% of the PM10 concentration and 8% of the OC concentration it is due to fungi spores, indicating its importance. Using receptor modeling it was identified six sources for PM10, the biomass burning resulting from industrial processes, the biomass burning resulting from vegetation burning, the fungal aerosol, the secondary formation aerosol, the soil resuspension and vehicular emission. These results are a breakthrough for research on bioaerosols in Brazil, since many of this analyzes had never been performed in urban areas in Brazil

Airborne infectious diseases remain a major global health threat. The primary vector for their transmission is coarse and fine droplets, known as bioaerosols, exhaled from infected individuals during natural respiratory maneuvers, such as breathing, coughing and sneezing. Unfortunately, our current knowledge of the mechanisms by which these exhaled bioaerosols are formed in the lungs is in its infancy. In particular, progress in this field has been hindered by the complex structure of the respiratory fluid and the resulting lack of understanding of the biophysical properties of the fluid. In this thesis, a series of in vitro mimetics of conducting airway mucus were developed to enable in depth studies of mucus properties and bioaerosol formation from mucus-like surfaces. These mucus mimetics overcome major limitations of currently available models by matching the primary chemical composition and key physical properties of the mimetic to that of native tracheal mucus. Three mimetics were chosen to facilitate the study of highly viscoelastic (diseased) mucus and non-diseased mucus under low shear conditions, such as breathing, or high shear conditions, such as cough. To study bioaerosol formation in vitro, an enhanced simulated cough machine (SCM) was developed to generate bioaerosols from mucus mimetic surfaces during cough. By controlling the temperature and relative humidity within the SCM, the detectability of bioaerosols generated from the mimetic surfaces was improved due to limited aerosol drying. The size distribution of the bioaerosols was multimodal, with four to five modes being observed for all surfaces probed. While varying the composition of surfactant at the air-mucus interface had a significant impact on surface viscoelastic properties, the size distribution of bioaerosols generated from these surfaces did not differ significantly. However, the ability to generate bioaerosols from different surfaces was highly dependent on surface properties of the mimetic, with highly viscoelastic surfaces generating bioaerosols in only half the experiments. This research will enhance our knowledge of bioaerosol formation in the respiratory tract and ultimately guide the development of alternative strategies to suppress bioaerosol formation.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-10-17T12:04:04Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Laureana Sobral - Dissertação PPGSHMA - Turma 2014.1.pdf: 2229448 bytes, checksum: e64e13ba35ae5b8c0a2b0c1183a66d3f (MD5) / Made available in DSpace on 2016-10-17T12:04:04Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Laureana Sobral - Dissertação PPGSHMA - Turma 2014.1.pdf: 2229448 bytes, checksum: e64e13ba35ae5b8c0a2b0c1183a66d3f (MD5) Previous issue date: 2016-07-12 / Fungos anemófilos são os principais contaminantes no ar de ambientes climatizados podendo promover agravos à saúde. Alguns produzem enzimas de interesse industrial e metabólitos à biotecnologia. O objetivo do trabalho foi avaliar a microbiota fúngica do ar de ambientes climatizados do Centro Acadêmico de Vitória/Universidade Federal de Pernambuco (CAV/UFPE) e seu potencial biotecnológico para produção enzimática e antimicrobiana. A amostra foi constituída por 82 ambientes climatizados e coletadas da área central dos ambientes através da técnica de sedimentação passiva sobre meios de cultura Ágar Extrato de Malte acrescido de Cloranfenicol e Ágar Aspergillus Flavus e Parasiticus para calcular a relação entre o ar interno e externo (I/E), quantificar colônias fúngicas e avaliar presença de cepas aflatoxigênicas confrontamento a legislação vigente da ANVISA. As colônias foram purificadas, identificada as espécies, preservadas e depositadas no acervo da Coleção de Culturas Micoteca URM da UFPE. Na determinação da atividade enzimática, discos de micélio puros foram repicados para a área central da placa de Petri contendo meios de cultura específicos para as enzimas lipase, amilase e protease e as reações foram constatadas respectivamente pela presença de cristais de sal de cálcio do ácido láurico, utilização de solução de iodo 0,1N e reação química, formando halo translúcido que determinou o potencial enzimático das colônias. Para realizar a atividade antimicrobiana 05 líquidos metabólicos foram testados para determinar a Concentrações Mínimas Inibitórias Bactericidas e Bacteriostáticas, os discos de micélios foram sepados do líquido metabólito para realização dos testes, utilizanddo cepas de bactérias Gram positivas e negativas, seguindo o método de diluição em caldo da norma técnica da ANVISA M7-A6 e revelados com cloreto de 2,3,5-trifeniltetrazólio. Foi observado crescimento fúngico e diversidade em todos os ambientes, a sala de aula apresentou o menor quantitativo de UFC/m3 (14) enquanto que o espaço farmácia viva, sala da CIPA e o gabinete docente, apresentaram a maior quantidade (290 UFC/m3). A relação I/E dos ambientes atendeu a legislação da ANVISA para qualidade do ar de ambientes climatizados (≤ 1,5), variando de 0,1 a 1,5, uma vez que a quantificação do ar externo foi de 188 UFC/m3 e nenhum ambiente apresentou fungos aflatoxigênicos. A frequência dos gêneros fúngicos identificados foram: Aspergillus (50%), Penicillium (21%), Talaromyces (14%), Curvularia e Paecilomyces (7% cada) e as espécies identificadas foram: Paecilomyces variotti, Penicillium fellutanum, Aspergillus flavus, Aspergillus parasiticus, Aspergillus sydowii, Talaromyces purpurogenus, Aspergillus japonicus, Penicillium oxalicum, Penicullium chrysogenum e Curvularia luneta. xiii Dentre as espécies testadas para atividade enzimática, Aspergillus sydowii apresentou melhor resultado para produção de amilase (6 mm), lipase (14 mm) e protease (5 mm). Outras duas espécies produziram as três enzimas (A. parasiticus e Penicillium fellutanum), porém com menor atividade. Apenas três isolados pertencentes a três espécies foram negativos para as 3 enzimas simultaneamente, a saber: Paecilomyces variotii, A. parasiticus lunata e Curvularia lunata. Em relação aos testes de atividade antibacteriana, três líquidos metabólicos apresentaram atividade contra todas as bactérias testadas (Paecilomyces variotii, Talaromyces purpurogenus e A. parasiticus). O líquido metabólico de Curvularia lunata não apresentou atividade antibacteriana contra as bactérias testadas e a espécie Talaromyces purpurogenus apresentou melhor atividade antibacteriana contra todas as bactérias testadas, chegando à concentração mínima inibitória de 125 μL/mL. Contudo, nenhum líquido metabólico apresentou ação bactericida, apenas bacteriostática. Concluindo que todos os ambientes apresentam biodiversidade de espécies fúngica e não foi encontrado nenhum fungo toxigênico ou patogênico ao homem, atendendo à legislação da ANVISA para ambientes climatizados, o maior potencial enzimático foi a lipolítica alcançada pelas espécies Aspergillus sydowii e Aspergillus parasiticus e os líquidos metabólicos das espécies Paecilomyces variotti, Talaromyces purpurogenus e Aspergillus parasiticus apresentaram atividade antimicrobiana contra todas as bactérias desafiadas, sendo a espécie Talaromyces purpurogenus a que obteve melhor resultado. Os fungos filamentosos do ar apresentam potencial para atividade enzimática e antimicrobiana. / Airborne fungi are the main contaminants in the air-conditioned environments can promote health problems. Some produce enzymes of industrial interest and metabolites biotechnology. The objective was to evaluate the fungal microbiota of the air-conditioned rooms of the Academic Center of Vitória/Federal University of Pernambuco (CAV/UFPE) and its biotechnological potential for enzyme and antimicrobial production. The sample consisted of 82 air-conditioned rooms and collected the central area of the environment through passive sedimentation technique on culture media Agar Malt Extract plus chloramphenicol and agar Aspergillus Flavus and parasiticus to calculate the relationship between indoor air and outdoor (I/E), quantify fungal colonies and evaluate the presence of aflatoxigênicas confrontation strains current legislation ANVISA. The colonies were purified, identified the species, preserved and deposited in the collection of the Culture Collection URM Culture Collection of the UFPE. In the determination of enzymatic activity, pure mycelial discs were transferred to the central area of the Petri plate containing specific culture media for lipase, amylase and protease and the reactions were detected respectively by the presence of calcium salt crystals of lauric acid, iodine solution using 0.1N and chemical reaction, forming translucent halo that determined the enzyme potential of the colonies. To achieve the antimicrobial activity 05 metabolic liquids were tested for minimal inhibitory concentrations bactericides and bacteriostatic the mycelial discs were sepados liquid metabolite for carrying out the tests, utilizanddo strains of positive and Gram negative bacteria, following the dilution method in bouillon the technical standard of ANVISA M7-A6 and stained with 2,3,5- triphenyltetrazolium. Growth was observed fungal and diversity in all environments, the classroom had the lowest quantity of UFC/m3 (14) while the space living pharmacy, CIPA room and the teacher's office, had the highest number (290 UFC/m3). The I/E ratio of environments picked ANVISA legislation for air quality in conditioned environment (≤ 1.5), ranging from 0.1 to 1.5, since the quantification of outdoor air was 188 UFC/m3 and no environment presented aflatoxigenic fungi. The frequency of the identified fungal genera are Aspergillus (50%), Penicillium (21%), Talaromyces (14%), Curvularia and Paecilomyces (7% each) and the identified species were Paecilomyces variotti, Penicillium fellutanum, Aspergillus flavus, Aspergillus parasiticus, Aspergillus sydowii, Talaromyces purpurogenus, Aspergillus japonicus, Penicillium oxalicum, Penicullium chrysogenum and Curvularia spyglass. Among the species tested for enzymatic activity, Aspergillus sydowii showed better results for amylase production (6 mm), lipase (14 mm) and protease (5 mm). Two other xv species produced the three enzymes (A. parasiticus and Penicillium fellutanum), but with less activity. Only three isolates belonging to three species were negative for the 3 enzymes simultaneously, namely Paecilomyces variotii, A. parasiticus lunata and Curvularia lunata. Regarding the antibacterial activity tests three metabolic liquid showed activity against all tested bacteria (Paecilomyces variotii, Talaromyces purpurogenus and A. parasiticus). Metabolic liquid Curvularia lunata showed no antibacterial activity against bacteria tested and Talaromyces purpurogenus species showed better antibacterial activity against all bacteria tested, reaching the minimum inhibitory concentration of 125 μL/mL. However, no metabolic liquid showed bactericidal action, only bacteriostatic. Concluding that all environments present biodiversity of fungal species and not found any toxigenic or pathogenic fungus to man, given the ANVISA legislation for climate-controlled environments, the highest enzyme potential was lipolytic achieved by the species Aspergillus sydowii and Aspergillus parasiticus and metabolic liquids species Paecilomyces variotti, Talaromyces purpurogenus and Aspergillus parasiticus presented antimicrobial activity against all challenged bacteria, Talaromyces purpurogenus species being that obtained better results. Filamentous fungi air show potential for enzyme and antimicrobial activity.

La qualité de l’air que nous respirons à l’intérieur des espaces clos est devenue une préoccupation environnementale majeure. Il est aujourd’hui avéré que la présence de microorganismes dans l’air intérieur peut avoir des conséquences néfastes sur la santé humaine. Les systèmes de ventilation, lorsqu’ils sont mal entretenus, peuvent être eux-mêmes la source de bioaérosols suite à la contamination de leurs filtres. Afin de pallier ce problème, des filtres photocatalytiques ont été développés et testés dans des centrales de traitement d’air (CTA). Ce projet de thèse s’intéresse donc au devenir des microorganismes aérosolisés dans des filtres photocatalytiques. La première étape a consisté à étudier les mécanismes de dégradation par photochimie et photocatalyse d’une bactérie modèle, Escherichia coli (E. coli), en suspension dans l’eau sous irradiation UV-A ou UV-C. Ces deux traitements sont plus efficaces sous UV-C et engendrent des dommages oxydatifs au niveau de la membrane externe et des acides nucléiques bactériens. Le dosage de sous-produits chimiques permet de conclure que la photocatalyse UV-C est plus efficace pour dégrader irréversiblement les bactéries. La seconde étape a été consacrée à l’étude de l’inactivation de spores d’Aspergillus niger et de bactéries E. coli dans les filtres Ahlstrom. Nos résultats confirment l’intérêt d’utiliser des filtres photocatalytiques dans les CTA en comparaison avec les filtres non photocatalytiques. L’utilisation de filtres à haute efficacité, afin d’optimiser le temps de contact entre les microorganismes et le dépôt de dioxyde de titane, permet d’augmenter l’efficacité de dégradation du procédé photocatalytique / Indoor Air Quality (IAQ) has become of particular concern from an environmental and health and safety point of view. It is now proven that the presence of microorganisms in indoor air can have hazardous consequences on human health. HVAC systems, when poorly maintained, can themselves become source of bioaerosols when their filters are contaminated. To tackle this issue, photocatalytic filters have been developed and tested within HVAC systems. The aim of this PhD thesis is to look at the behaviour of microorganisms aerosolized through photocatalytic filters. The first step consisted of analysing the photochemistry and photocatalysis degradation mechanisms of a model bacterium, Escherichia coli (E. coli), in suspension in water under UV-A or UV-C radiation. These two treatments were proven to be more efficient under UV-C radiation and led to oxidative damages to outer membrane and nucleic acids. The measurement of chemical by-products allowed us to conclude that UV-C photocatalysis is more effective in damaging bacteria in an irreversible manner. The second step focused on the analysis of the inactivation of Aspergillus niger spores and E. coli bacteria within Ahlstrom filters. Our results confirm the benefits of using photocatalytic filters in HVAC systems instead of non-photocatalytic filters. The use of high efficiency filters to optimise the contact between the microorganisms and the titanium dioxide deposit, allows an increase in the damaging efficiency of the photocatalytic process

En passant près de 90% de son temps dans les espaces clos, l'Homme est exposé à des polluants particulaires de diverses natures d'origines exogène et endogène au bâtiment, pour lesquels aucune valeur guide n'est disponible. Parmi ces polluants figurent les particules biologiques et notamment les spores fongiques, particules vivantes les plus nombreuses et les plus diversifiées de l'air que nous respirons (Nolard, 1997). Ubiquitaire et délétère, la pollution particulaire fongique est mise en cause dans la survenue de nombreuses pathologies parmi lesquelles se trouvent les maladies immuno-allergiques. Dans le cadre de la surveillance de la qualité micro-biologique de l'air des espaces clos, cette thèse vise à fournir les premiers éléments de conception d'un outil individuel de diagnostic dédié à l'évaluation de l'exposition des occupants aux aérobiocontaminants allergéniques en s'intéressant plus particulièrement à la pollution fongique aéroportée. Cette recherche repose sur les expertises techniques et scientifiques du CSTB, de l'ESIEE et de l'Université Paris-Est en matière de détection fongique, de miniaturisation d'instruments de mesure via les micro technologies et de physique des aérosols. Se faisant, ces travaux cristallisent autour d'une architecture système reposant sur trois axes : la capture et sélection des particules selon leurs propriétés physico-chimiques de surface, la quantification de la masse des particules et l'identification de la nature des particules à l'aide d'une analyse chimique. Ces axes correspondent à autant de thématiques abordées au cours de ces travaux de thèse. Ainsi, la première a consisté à étudier l'adhérence des conidies aux surfaces afin de mieux cerner les déterminants de ce phénomène physique et évaluer les énergies mises en jeu. Les résultats subséquents ont permis, lors d'une deuxième phase de travail, de dimensionner une microbalance en silicium de type MEMS en replaçant les problématiques liées aux interactions particules-résonateurs au cœur du débat. Par ce biais, l'enjeu a été de lever certains verrous scientifiques constatés dans la littérature, telles des sensibilités non-uniformes sur toute la surface des dispositifs de mesure ou encore des réponses en fréquence non linéaires avec la masse déposée. Une telle approche a en outre permis d'évaluer les performances attendues pour de tels capteurs. Enfin, le dernier aspect de cette recherche a porté sur l'identification des particules aéroportées biologiques par voie chimique en combinant pyrolyse des entités biologiques d'intérêt, chromatographie en phase gazeuse et spectroscopie de masse (Py-CPG/MS). A cette occasion, un travail collaboratif engagé avec le Réseau National de Surveillance Aérobiologique a permis d'éprouver la solution technologique et la méthodologie employées puisqu'une seconde catégorie de particules modèles a alors été considérée : les pollens. L'analyse des composés organiques volatiles issus de l'analyse Py-CPG/MS des micromycètes et des pollens a permis de démontrer l'existence d'une signature chimique spécifique de l'origine biologique des particules. Suite à cela, il a été possible d'établir diverses listes de traceurs chimiques caractéristiques des phyla voire des espèces des différents contaminants étudiés. La pertinence de ces marqueurs a alors été éprouvée lors d'un essai in situ / Nowadays people pass 90% of their time in closed spaces, and in consequence are exposed to indoor and outdoor particulate matter for which no reference value is available. These pollutants include biological particles and in particular fungal spores, the most numerous living particles and most diverse on the air we breathe (Nolard, 1997). Ubiquitous and harmful, fungal particulate pollution is implicated in the occurrence of many diseases including immuno-allergic diseases. In the context of the monitoring of the microbiological quality of air in indoor spaces, this thesis aims to provide first design elements of an individual diagnostic device dedicated to the exposure assessment of allergenic bio-contaminants focusing in particular on airborne fungal pollution. This research relies on the technical and scientific expertise of CSTB, ESIEE Paris and Université Paris-Est for fungal detection, miniaturization of measurement instrumentation and aerosol physics. Thus, this work is built around a system architecture based on three main elements: the capture and selection of particles according to their surface physical and chemical properties, the particles mass quantification and the identification of the nature of the particle using chemical analysis. These elements relate to many topics covered during the thesis work. In this way, the first topic consists in studying the adhesion of conidia to surfaces to better understand the determinants of this physical phenomenon and evaluate the energies involved. Subsequent results were used during a second stage of this work, to design a MEMS-type silicon microbalance considering the particle-resonator interaction. By this mean the issue was to solve some scientific challenges identified in the literature, such non-uniform sensitivity over the entire device surface or nonlinear frequency responses due to the added mass. Such an approach has also allowed evaluating the performance expected for such sensors. The last aspect of this research focused on the identification of biological airborne particles chemically combining pyrolysis of biological entities of interest, and gas chromatography and mass spectrometry (Py-GC/MS). On this occasion, a collaborative work engaged with the "Réseau National de Surveillance Aérobiologique" allowed to experience the technological solution and our methodology since another class of particles was considered: pollens. The analysis of volatile organic compounds obtained from Py-GC/MS characterization of micro-fungi and pollens demonstrated the existence of a specific chemical signature for each biological particle class. Thereafter, it was then possible to establish a variety of chemical markers lists for phyla and different species of the contaminants studied. The relevance of these markers has been further tested in an in-situ assay