Devido à ampla utilização do cádmio nas indústrias, muitos solos encontram-se contaminados. Vários estudos com bioindicadores estão sendo realizados para detectar cádmio em solos, sendo a maioria com minhocas (Eisenia fetida) e plantas, contudo estes testes são laboriosos e longos. Os micro-organismos tem se mostrado uma alternativa promissora para esta aplicação, pois são de fácil cultivo e manutenção, além de apresentarem uma rápida resposta às alterações ambientais. Desta forma, o objetivo deste trabalho foi utilizar diferentes gêneros de leveduras como bioindicadores de cádmio no solo. Para tanto foram avaliadas dezoito linhagens de leveduras, das quais foram selecionadas três quanto à sensibilidade na presença de cádmio. A levedura Torulopsis utilis var. thermophilus (IZ214) apresentou alta sensibilidade, a Candida utilis (IZ300) sensibilidade mediana e Saccharomyces cerevisiae (Pedra) a menor sensibilidade. O crescimento foi avaliado por espectrometria, através de leituras de D.O. e pela técnica do Trifeniltetrazólio (TTC) para avaliação da viabilidade celular. O uso do TTC mostrouse mais adequado nos bioensaios com leveduras evidenciando as células metabolicamente ativas. Foi avaliado o crescimento das leveduras em extratos de solo com cádmio obtidos pelo extrator CaCl2 0,01 mol L-1. Observou-se que o crescimento das leveduras na maioria dos tratamentos foi igual ou superior ao controle. Isso ocorreu devido à adsorção do cádmio à matéria orgânica e outros elementos. Para comprovar a adsorção do cádmio à matéria orgânica, foi realizado ensaio com a levedura S. cerevisiae (Pedra) onde ao extrato de solo obtido foi adicionado solução de Cd2+. Através da leitura de D.O. foi observada uma taxa de inibição de 75% para 100 mg L-1 de cádmio e, pela técnica do TTC, a taxa de inibição de 100% ocorreu na concentração de 44 mg L-1. Ensaios com sementes foram realizados para avaliar a germinação e crescimento na presença de cádmio. Sementes de Cucumis sativus foram expostas à solução de Cd2+, ocorrendo 100% de germinação e apresentando DL50 de 7,33 mg L-1. Este resultado foi maior comparado aos obtidos para as leveduras avaliadas, as quais apresentaram uma DL50 de 0,89 mg L-1 para a levedura T. utilis (IZ214) e DL50 de 1,86 mg L-1 para a S. cerevisiae (Pedra). As sementes de Zea mays foram expostas ao extrato de solo com cádmio, onde foi obtido uma DL50 de 33,9 ± 3,0 mg L-1 e, nas mesmas condições para a levedura S. cerevisae (Pedra), a DL50 foi de 29,8 mg L-1. Os resultados obtidos mostraram uma proximidade de valores nos ensaios com extrato de solo com cádmio entre a semente de milho e levedura. Com isso, foi possível concluir que as leveduras são sensíveis ao cádmio e podem ser uma nova opção de bioindicadores. Além disso, as leveduras e principalmente a S. cerevisiae possuem um alto grau de homologia com os eucariotos superiores, permitindo assim o estudo de aspectos de toxicidade relevantes a biologia humana. / Due to extensive use of cadmium in industries, many soils are contaminated. Several studies are being conducted with biomarkers to detect cadmium in soils, mostly with earthworms (Eisenia fetida) and plants, however, these tests are laborious and lengthy. Micro-organisms have been presented as an alternative to this application, as they are easy to grow and maintain, and offer a rapid response to environmental changes. Thus, the aim of this study was to use different kinds of yeasts as indicators of cadmium in soil. Eighteen yeasts strains were evaluated, from which three were selected for their sensitivity in the presence of cadmium. The yeast Torulopsis utilis var. thermophilus (IZ214), showed high sensitivity, Candida utilis (IZ300), showed a median sensitivity and Saccharomyces cerevisiae (Ale Yeast) a showed lower sensitivity. Yeasts growth was assessed through spectrophotometry, by reading O.D. and by the technique Triphenyltetrazolium (TTC) to assess cell viability. The use of TTC was more appropriate as an indicator of cell viability in yeast bioassays showing metabolically active cells. We evaluated the growth of yeasts in soil extracts with cadmium obtained by extraction with CaCl2 0.01 mol L-1. It was observed that the growth of the yeast on most of the treatments was equal or superior to the control. This was due to the adsorption of cadmium to organic matter and other elements. To prove the adsorption of cadmium to organic matter, an essay was performed with the yeast S. cerevisiae (Ale Yeast), in which the soil extract solution obtained was added with Cd2+. By reading the O.D. it was observed an inhibition rate of 75% for 100 mg L- 1 cadmium and, by the TTC technique, the highest rate of 100% inhibition occurred at a concentration of 44 mg L-1. Seeds trials were conducted to evaluate germination and growth. Cucumis sativus seeds were exposed to a solution of Cd2+, resulting in 100% of germination and showing a LD50 of 7.33 mg L-1, which was greater than the one measured for yeasts, who had an LD50 of 0.89 mg L-1 to the yeast T. utilis (IZ214) and LD50 of 1.86 mg L-1 for S. cerevisiae (Ale Yeast). The seeds of Zea mays were exposed to soil extract containing cadmium, in which it was noticed a LD50 of 33.9 ± 3.0 mg L-1 and, by having the same conditions for the yeast S. cerevisiae (Ale Yeast), the LD50 was of 29.8 mg L-1. The results showed similar values in tests with soil extract with cadmium comparing the corn seed and the yeast. Thus, it was concluded that the yeasts are sensitive to cadmium and may be a new option bioindicators. Also, yeasts, in special S. cerevisiae, have a high degree of homology with higher eukaryotes, therefore allowing the study of relevant toxicity aspects in human biology.

Este trabalho objetivou estudar a possibilidade de se utilizar líquen como bioindicador de poluição atmosférica para regiões impactadas por radionuclídeos, metais e elementos terras raras. Foram escolhidas como áreas de estudo a região de Pirapora do Bom Jesus, onde se localiza uma indústria de produção de estanho e chumbo metálicos e a região de Cubatão, onde se localiza o complexo industrial de produção de fertilizantes fosfatados. As duas instalações escolhidas são consideradas como indústrias TENORM - Technologically Enhanced Naturally Occurring Radioactive Material, pois podem ocasionar um aumento significativo na concentração dos radionuclídeos naturais nas várias etapas do processo industrial, ocasionando um aumento potencial de exposição à radiação natural em produtos, subprodutos e resíduos gerados. Para tanto foram analisados os radionuclídeos 238U, 226Ra, 210Pb, 232Th e 228Ra, os elementos terras raras e metais em amostras de matéria prima e resíduo das instalações, liquens e solos na área de influência. As amostras de líquen e solo foram analisadas para a determinação de urânio, tório, elementos terras raras e metais por análise por ativação com nêutrons. Os radionuclídeos 226Ra, 228Ra e 210Pb em amostras de solo foram determinados por espectrometria gama, e em amostras de liquens por separação radioquímica e contagem alfa e beta total em contador proporcional de fluxo de gás constante. As concentrações de 238U (de 19 a 473 Bq kg-1), 226Ra (de 21 a 265 Bq kg-1), 210Pb (de 401 a 1461 Bq kg-1), 232Th (de 15 a 574 Bq kg-1), 228Ra (de 176 a 535 Bq kg-1), elementos terras raras, Hf e Ta determinadas nas amostras de liquens ao redor da indústria de produção de estanho e chumbo metálicos mostram que todos estes elementos se concentram significativamente e, portanto, os liquens fornecem uma assinatura da contaminação atmosférica. Os resultados obtidos nas amostras de liquens da região de Cubatão mostram uma assinatura mais evidente do radionuclídeo 210Pb, proveniente de todo parque industrial da região. Os resultados obtidos no presente estudo mostraram que os liquens podem ser usados como bioindicadores de poluição atmosférica por radionuclídeos e elementos traço. / The main aim of this thesis is to study the possibility of using lichen as bioincicator of atmospheric pollution in regions contaminated by radionuclides, metals and rare earth elements. Two regions were chosen, one in Pirapora do Bom Jesus, where a tin and lead industry is located, and a second one in Cubatão, where a phosphate fertilizer industrial complex is located. The two industries chosen are considered as TENORM - Technologically Enhanced Naturally Occurring Radioactive Material, since they can cause a significant increase in the natural radionuclides concentration in the industrial process, and consequently a potential increase in the radiation exposure in products, byproducts and residue. To achieve this aim, the radionuclides 238U, 226Ra, 210Pb, 232Th and 228Ra, rare earth elements and metals were analyzed in samples of raw material an residue from the installations, lichens and soils. Lichens and soil samples were analyzed by instrumental neutron activation analysis for the determination of uranium, thorium, rare earth elements and metals. The radionuclides 226Ra, 228Ra and 210Pb in soil samples were determined by gamma spectrometry and in lichen sample by radiochemical separation and gross alpha and beta counting on a gas flow proportional counter. The concentrations of 238U (from 19 to 473 Bq kg-1), 226Ra (from 21 to 265 Bq kg-1), 210Pb (from 401 to 1461 Bq kg-1), 232Th (from 15 to 574 Bq kg-1), 228Ra (from 176 to 535 Bq kg-1), rare earth elements, Hf and Ta determined in lichen samples around the tin and lead industry show an enrichment in these elements. Therefore, the lichens can be used as a fingerprint of the atmospheric contamination. The results obtained for the lichen samples, in the Cubatão region, present a fingerprint mainly of 210Pb, from industries of the region. The results obtained in this study showed that the lichens can be used as bioindicators of atmospheric pollution by radionuclides and trace elements.

Devido à ampla utilização do cádmio nas indústrias, muitos solos encontram-se contaminados. Vários estudos com bioindicadores estão sendo realizados para detectar cádmio em solos, sendo a maioria com minhocas (Eisenia fetida) e plantas, contudo estes testes são laboriosos e longos. Os micro-organismos tem se mostrado uma alternativa promissora para esta aplicação, pois são de fácil cultivo e manutenção, além de apresentarem uma rápida resposta às alterações ambientais. Desta forma, o objetivo deste trabalho foi utilizar diferentes gêneros de leveduras como bioindicadores de cádmio no solo. Para tanto foram avaliadas dezoito linhagens de leveduras, das quais foram selecionadas três quanto à sensibilidade na presença de cádmio. A levedura Torulopsis utilis var. thermophilus (IZ214) apresentou alta sensibilidade, a Candida utilis (IZ300) sensibilidade mediana e Saccharomyces cerevisiae (Pedra) a menor sensibilidade. O crescimento foi avaliado por espectrometria, através de leituras de D.O. e pela técnica do Trifeniltetrazólio (TTC) para avaliação da viabilidade celular. O uso do TTC mostrouse mais adequado nos bioensaios com leveduras evidenciando as células metabolicamente ativas. Foi avaliado o crescimento das leveduras em extratos de solo com cádmio obtidos pelo extrator CaCl2 0,01 mol L-1. Observou-se que o crescimento das leveduras na maioria dos tratamentos foi igual ou superior ao controle. Isso ocorreu devido à adsorção do cádmio à matéria orgânica e outros elementos. Para comprovar a adsorção do cádmio à matéria orgânica, foi realizado ensaio com a levedura S. cerevisiae (Pedra) onde ao extrato de solo obtido foi adicionado solução de Cd2+. Através da leitura de D.O. foi observada uma taxa de inibição de 75% para 100 mg L-1 de cádmio e, pela técnica do TTC, a taxa de inibição de 100% ocorreu na concentração de 44 mg L-1. Ensaios com sementes foram realizados para avaliar a germinação e crescimento na presença de cádmio. Sementes de Cucumis sativus foram expostas à solução de Cd2+, ocorrendo 100% de germinação e apresentando DL50 de 7,33 mg L-1. Este resultado foi maior comparado aos obtidos para as leveduras avaliadas, as quais apresentaram uma DL50 de 0,89 mg L-1 para a levedura T. utilis (IZ214) e DL50 de 1,86 mg L-1 para a S. cerevisiae (Pedra). As sementes de Zea mays foram expostas ao extrato de solo com cádmio, onde foi obtido uma DL50 de 33,9 ± 3,0 mg L-1 e, nas mesmas condições para a levedura S. cerevisae (Pedra), a DL50 foi de 29,8 mg L-1. Os resultados obtidos mostraram uma proximidade de valores nos ensaios com extrato de solo com cádmio entre a semente de milho e levedura. Com isso, foi possível concluir que as leveduras são sensíveis ao cádmio e podem ser uma nova opção de bioindicadores. Além disso, as leveduras e principalmente a S. cerevisiae possuem um alto grau de homologia com os eucariotos superiores, permitindo assim o estudo de aspectos de toxicidade relevantes a biologia humana. / Due to extensive use of cadmium in industries, many soils are contaminated. Several studies are being conducted with biomarkers to detect cadmium in soils, mostly with earthworms (Eisenia fetida) and plants, however, these tests are laborious and lengthy. Micro-organisms have been presented as an alternative to this application, as they are easy to grow and maintain, and offer a rapid response to environmental changes. Thus, the aim of this study was to use different kinds of yeasts as indicators of cadmium in soil. Eighteen yeasts strains were evaluated, from which three were selected for their sensitivity in the presence of cadmium. The yeast Torulopsis utilis var. thermophilus (IZ214), showed high sensitivity, Candida utilis (IZ300), showed a median sensitivity and Saccharomyces cerevisiae (Ale Yeast) a showed lower sensitivity. Yeasts growth was assessed through spectrophotometry, by reading O.D. and by the technique Triphenyltetrazolium (TTC) to assess cell viability. The use of TTC was more appropriate as an indicator of cell viability in yeast bioassays showing metabolically active cells. We evaluated the growth of yeasts in soil extracts with cadmium obtained by extraction with CaCl2 0.01 mol L-1. It was observed that the growth of the yeast on most of the treatments was equal or superior to the control. This was due to the adsorption of cadmium to organic matter and other elements. To prove the adsorption of cadmium to organic matter, an essay was performed with the yeast S. cerevisiae (Ale Yeast), in which the soil extract solution obtained was added with Cd2+. By reading the O.D. it was observed an inhibition rate of 75% for 100 mg L- 1 cadmium and, by the TTC technique, the highest rate of 100% inhibition occurred at a concentration of 44 mg L-1. Seeds trials were conducted to evaluate germination and growth. Cucumis sativus seeds were exposed to a solution of Cd2+, resulting in 100% of germination and showing a LD50 of 7.33 mg L-1, which was greater than the one measured for yeasts, who had an LD50 of 0.89 mg L-1 to the yeast T. utilis (IZ214) and LD50 of 1.86 mg L-1 for S. cerevisiae (Ale Yeast). The seeds of Zea mays were exposed to soil extract containing cadmium, in which it was noticed a LD50 of 33.9 ± 3.0 mg L-1 and, by having the same conditions for the yeast S. cerevisiae (Ale Yeast), the LD50 was of 29.8 mg L-1. The results showed similar values in tests with soil extract with cadmium comparing the corn seed and the yeast. Thus, it was concluded that the yeasts are sensitive to cadmium and may be a new option bioindicators. Also, yeasts, in special S. cerevisiae, have a high degree of homology with higher eukaryotes, therefore allowing the study of relevant toxicity aspects in human biology.

Coral reefs are threatened in Florida and worldwide. Successful resource management requires rapid identification of anthropogenic sources of stress before they affect the reef community. I tested a multi-scale approach for assessing reef condition at seven reefs within the Florida Keys National Marine Sanctuary and Biscayne National Park between 2001 and 2003. I examined multiple environmental parameters to identify potential sources of stress. I utilized the Atlantic and Gulf Rapid Reef Assessment Biotic Reef Index to assess benthic community structure and an indicator species of Foraminifera (Amphistegina gibbosa) to determine if environmental conditions were suitable for calcareous organisms that host algal endosymbionts. Small tissue samples were extracted from colonies of Montastraea annularis species complex to assay a suite of cellular biomarkers to elucidate possible mechanisms of the coral stress response. I monitored regeneration rates of the resultant lesions to determine if the coral colonies were capable of recovering from damage. Multivariate data analyses indicated that corals at all study sites were experiencing stress with different degrees of response and decline. On reefs with coarse grain sediments that are adjacent to an intact mangrove shoreline, the Cellular Diagnostic System indicated that corals were responding to a xenobiotic stress but appeared to be compensating as evidenced by consistently high lesion regeneration rates, a high percentage of healed lesions, low coral mortality and high abundances of A. gibbosa. On reefs with silt-sized sediments adjacent to developed coastlines, corals also were responding to xenobiotic stresses, but were negatively affected as evidenced by low regeneration rates, a low percentage of healed lesions, high coral mortality, and low abundances of A. gibbosa. Corals at an 18 m offshore site exhibited abnormally low biomarker levels and some died during the study, indicating that sampled colonies were incapable of upregulating necessary protective proteins. Further research will be required to determine stressor sources. This study demonstrates that a multiple-indicator approach, spanning scales from cellular to community, can provide marine resource managers with data linking decline of coral populations to specific environmental conditions and events, thereby providing potential for early detection of stressors allowing for preventive management.

Studies of larger symbiont-bearing foraminifers on reefs have revealed their potential as indicators of environmental stress because of their physiological analogies to corals (dependence on algal symbionts for growth and calcification) and relatively short life cycle (a few months to 2 years or more). The purpose of this study is to report distribution patterns and population densities of larger benthic foraminifers (LBF) of the Florida reef tract, specifically reporting abundance data collected from offshore (1995-2000, 2006, 2007) and patch reefs (1996, 2006, 2007). Six years of quarterly data collected from two offshore reefs, Conch (10, 18 and 30m) and Tennessee (8 and 20m), revealed that LBF assemblages primarily varied with habitat depth, in turn reflecting available light and water motion.These assemblages were dominated by Amphistegina gibbosa d'Orbigny and Laevipeneroplis proteus d'Orbigny, which tended to occur together, making up ~40-50% of the assemblages and up to 80% at the Tennessee 20m site. Both overall abundance and evenness of the LBF assemblage structure exhibited the greatest variability at shallower depths. Evenness was inversely related to densities of A. gibbosa, which were typically higher at depth keeping evenness below 0.5. Across the Keys, region (location along the reef tract), reef type (offshore shallow, deep or patch reefs) and symbiont type strongly influenced LBF assemblage dynamics. Upper Keys sites shared the highest degree of inter-region similarity among assemblages (73%), while Biscayne National Park (BNP) and lower Keys sites had the lowest similarity (~60%). This likely reflects the greater variability of habitats found in the latter areas, mainly patch reefs.Chlorophyte-bearers were typically more abundant in shallower turbid waters, with diatom-bearers more abundant at depth. Additionally, I observed a significant two-fold decrease in the proportion of chlorophyte-bearers in the middle Keys likely due to light-limitation by turbid Florida Bay outflow. Finally, data comparisons revealed an inverse relationship between LBF abundances and percent coral cover. Coral cover (2005) was staggeringly low on offshore reefs (5%), but was significantly higher on nearshore patch reefs (12%). Contrastingly, LBF species showed either no difference in abundance between reef types or a greater abundance on offshore reefs.

Ένα από τα μεγαλύτερα περιβαλλοντικά προβλήματα που αντιμετωπίζει όχι μόνο ο Νομός Αχαΐας, αλλά και πολλές άλλες περιοχές της χώρας μας, είναι η υποβάθμιση του περιβάλλοντος από μια σειρά αποβλήτων αγροτοβιομηχανιών που υπάρχουν στην ελληνική ύπαιθρο. Μεταξύ των αγροτοβιομηχανιών σημαντική θέση κατέχουν οι μονάδες επεξεργασίας γάλακτος και παραγωγής τυριού. Παρά το ότι η παραγωγή γάλακτος και γαλακτοκομικών προϊόντων είναι σχετικά περιορισμένη στο Νομό Αχαΐας, ωστόσο η διάσπαρτη κατανομή τους σε συνδυασμό με τις κρατούσες συνθήκες λειτουργίας τους συμβάλλει σημαντικά στη ρύπανση του περιβάλλοντος της περιοχής. Είναι γεγονός ότι οι περισσότερες αγροτοβιομηχανίες, όπως τα τυροκομεία, λειτουργούν σε περιοδική βάση αφού τα προϊόντα τους είναι εποχικά, τα εργοστάσια είναι διασκορπισμένα σε μεγάλες αποστάσεις και οι μονάδες είναι σχετικά μικρές. Τα απόβλητα που παράγονται από τις τυροκομικές μονάδες έχουν υψηλό οργανικό φορτίο και από προηγούμενη έρευνα έχει προσδιοριστεί ότι είναι τοξικά και αποτελούν αιτία υποβάθμισης των υδάτινων οικοσυστημάτων. Η παρούσα μελέτη έχει ως βασικό σκοπό την εκτίμηση της τοξικότητας των αποβλήτων που προέρχονται από τυροκομεία μετά από την επεξεργασία τους για παραγωγή βιαερίου (υδρογόνου και μεθανίου) σε αντιδραστήρες αναερόβιας χώνευσης (Η2-CSTR και CH4-CSTR αντίστοιχα), με σκοπό την συνολική εκτίμηση των επιπτώσεών τους στο υδάτινο οικοσύστημα. Αρχικά εκτιμήθηκε ο οικολογικός κίνδυνος από την απόρριψη των αποβλήτων μιας τυροκομικής μονάδας στο ποτάμι του Βουραϊκού. Τα δεδομένα που συγκεντρώθηκαν δίνουν ετήσια πρόβλεψη για κρίσιμα επίπεδα οικολογικού κινδύνου σε εποχές όπως είναι το καλοκαίρι όπου η ροή νερού στο ποτάμι είναι μικρή, και για μικρότερο, αλλά και πάλι αξιοσημείωτο επίπεδο κινδύνου κατά το φθινόπωρο. Οι βροχοπτώσεις είναι λογικό να δημιουργούν μεγαλύτερη αραίωση του αποβλήτου στο νερό γεγονός που προκαλέι μείωση των επιπέδων κινδύνου. Η συνδυασμένη ανάλυση των δεδομένων απέδειξε ότι εξαιρετικά μικρή συγκέντρωση του αποβλήτου, μικρότερη του 0,064%, είναι προϋπόθεση για να αποφευχθεί ο οικολογικός κίνδυνος για την υδρόβια πανίδα. Όμως, η παρούσα οικολογική κατάσταση του οικοσυστήματος απέχει πολύ από το να χαρακτηριστεί ως βέλτιστη, μιας και η απόρριψη ανεπεξέργαστων αποβλήτων στο νερό προκαλεί συγκεντρώσεις 5 φορές μεγαλύτερες από το ανωτέρω όριο, πλησιάζοντας το 0,32% κατά τη διάρκεια του καλοκαιριού. Ο έλεγχος τοξικότητας επιτεύχθηκε με τη χρήση βιοδεικτών από δύο τροφικά επίπεδα (ασπόνδυλα και ψάρια του γλυκού νερού). Συγκεκριμένα, στην πρώτη περίπτωση χρησιμοποιήθηκαν δύο μακροασπόνδυλα (Daphnia magna και Thamnocephalus platyurus) με τη μορφή των μικροβιοτέστ Thamnotoxkit F και Daphtoxkit FTM magna, τα οποία επελέγησαν λόγω της αξιοπιστίας τους, της σχετικής ευκολίας τους στη χρήση, του χαμηλού κόστους και της προοπτικής τους να χρησιμοποιηθούν από μη εξειδικευμένο προσωπικό για την παρακολούθηση των επιπτώσεων των συγκεκριμένων αποβλήτων. Στη δεύτερη περίπτωση έγινε ο έλεγχος της τοξικότητας με έμβρυα zebrafish (Danio rerio) με βάση τα πρωτόκολλα των σχετικών ISO και της EPA. Λήφθηκαν συνολικά 109 δείγματα (4 διπλά δείγματα τυρόγαλου, 55 διπλά δείγματα από τον αντιδραστήρα παραγωγής υδρογόνου και 50 διπλά δείγματα από τον αντιδραστήρα παραγωγής μεθανίου). Στα τεστ τοξικότητας που εφαρμόστηκαν (Thamnotoxkit F, Daphtoxkit FTM magna και zebrafish), υπολογίσθηκαν τα L(Ε)C50 24h και 48h σύμφωνα με τα πρωτόκολλα εργασίας. Ευρέθη ότι το μεν Thamnocephalus platyurus είχε μέση τιμή LC50 ίση με 0,76 για τα δείγματα από τον αντιδραστήρα Η2-CSTR και 1,33 για τα δείγματα από τον αντιδραστήρα CH4-CSTR, οι αντίστοιχες τιμές για την Daphnia magna στις 48 ώρες ήταν 1,82 και 2,26 ενώ για το zebrafish 0,88 και 0,95 στο ίδιο διάστημα. Οι τιμές που προέκυψαν από τους τρεις ελέγχους κατατάσσουν τα απόβλητα από «πολύ τοξικά» έως «εξαιρετικά τοξικά». Από τη συσχέτιση των φυσικοχημικών παραμέτρων με τα L(Ε)C50 προκύπτει ότι για τα δείγματα του Η2-CSTR υπάρχει θετική συσχέτιση των αμμωνιακών, νιτρικών και νιτρωδών ιόντων με το Thamnocephalus platyurus (R= 0,368, R=0,442 και R=0,362 αντίστοιχα) και για τα δείγματα του CH4-CSTR συσχέτιση υπάρχει μόνο με τα ολικά διαλυμένα στερεά (R=0,860). Για το zebrafish, υπάρχει συσχέτιση με τα φωσφορικά (R= 0.542) και με τα αμμωνιακά ιόντα (R=0,562) για τα δείγματα του Η2-CSTR, ενώ για τα δείγματα του CH4-CSTR με τα φωσφορικά (R=0,963) και τα νιτρώδη ιόντα (R= 0,960). Η Daphnia magna δε δείχνει καμία σημαντική συσχέτιση, με τη μεγαλύτερη εξ αυτών να παρατηρείται με τα αμμωνιακά ιόντα (R= 0,316) μόνο για τα δείγματα του Η2-CSTR. Σύμφωνα με τα ανωτέρω αποτελέσματα τεκμηριώνεται ότι το Thamnocephalus platyurus και το zebrafish είναι οι πιο ευαίσθητοι και επομένως οι πλέον κατάλληλοι οργανισμοί για την εκτίμηση της τοξικότητας των συγκεκριμένων αποβλήτων. Η παρατήρηση σκελετικών δυσμορφιών έστω και σε μικρό ποσοστό δειγμάτων του zebrafish, οδήγησε στην ανίχνευση βαρέων μετάλλων σε όλα τα επεξεργασμένα δείγματα δεδομένου ότι και αυτά έχουν ενοχοποιηθεί για την πρόκληση αυτών. Συγκεκριμένα ανιχνεύθηκαν χρώμιο, μαγγάνιο, ψευδάργυρος και μόλυβδος, όμως ο ρόλος καθενός από αυτά ή και συνεργιστικά όλων αυτών στις παρατηρηθείσες δυσμορφίες δε μπορεί να διερευνηθεί πλήρως στα πλάισια αυτής της μελέτης και για τούτο θα απαιτηθεί περαιτέρω έρευνα. Συνοπτικά αποδεικνύεται ότι όλα τα επεξεργασμένα δείγματα και με τις δύο μεθόδους ήσαν τοξικά και όχι περιβαλλοντικά ασφαλή για άμεση διάθεση σε υδάτινο αποδέκτη χωρίς περαιτέρω επεξεργασία για την απομάκρυνση των υπόλοιπων επιβαρυντικών παραγόντων όπως ο φώσφορος και οι ενώσεις του αζώτου. Τέλος, θα πρέπει να σημειωθεί ότι η παρούσα μελέτη είναι η πρώτη μελέτη εκτίμησης της τοξικότητας των αποβλήτων των τυροκομικών μονάδων μετά από την επεξεργασία του για παραγωγή βιοαερίου που γίνεται στη χώρα μας, αλλά και γενικότερα δεν υπάρχει ανάλογη αναφορά στη διεθνή βιβλιογραφία. Δεδομένου ότι η προαναφερθείσα επεξεργασία αποτελεί μια καινοτομία συνδυασμένης μεθοδολογίας που αποβλέπει αφενός στην εξυγίασνη του αποβλήτου και αφετέρου στην παραγωγή ωφέλιμου παραπροϊόντος όπως είναι το βιοαέριο, τα αποτελέσματα αυτής της έρευνας θα μπορούσαν να αποτελέσουν ένα αξαιρετικά χρήσιμο εργαλείο, αφενός στους επιστήμονες που ασχολούνται με την επεξεργασία των αποβλήτων ώστε να αναζητήσουν τρόπους περαιτέρω αποφόρτισης αυτών από τους εναπομείναντες τοξικούς παράγοντες και αφετέρου στην πολιτεία για τη συστηματική παρακολούθηση και τον έλεγχο των συγκεκριμένων μονάδων στα πλαίσια εφαρμογής ορθής περιβαλλοντικής πολιτικής. / One of the major environmental hazard not only for the Achaia Prefecture, but also for many other regions of the Greek area has to deal with agro-industrial effluents which outfall to the countryside. Among the agro-industries, the units of milk treatment and cheese production are concerned as such of great environmental interest. Despite that the production of milk and dairy products is relatively limited in Achaia Prefecture, their scattered installation and their unlegal operation contribute considerably in the environmental pollution of the area. The most dairy units operate in periodical base, they are scattered in the countryside while they are relatively assessed as small units. Their effluents that have been considered as toxic. The basic aim of the present study is to estimate the toxicity of cheese whey effluents from dairy unit after their treatment for biogas production (hydrogen and methane) in anaerobic digestion reactors (H2-CSTR and CH4-CSTR respectively), aiming at the total estimation of their repercussions in the aquatic ecosystem. Initially, the ecological risk from the rejection of cheese-whey effluents in the river of Vouraikos has been estimated. The data drived to an annual forecast which gives critical levels of ecological risk during summertime, and remarkable level of risk during autumn. Integrated analysis proved that effluents concentration of the river smaller than 0,064% is prerequired in order to avoid the ecological risk of the aquatic fauna, while the present ecological situation of ecosystem is far from optimal, as the rejection of the untreated effluents into the water leads to concentrations 5 times higher than the above limit, approaching 0,32% during summertime. The toxicity estimation was achieved with the use of bioindicators from two trophic levels (freshwater invertebrates and fish): Daphnia magna and Thamnocephalus platyurus in the form of mikrobiotests Thamnotoxkit F and Daphtoxkit FTM magna, and the embryos of zebrafish (Danio rerio) according to the protocols of ISO and EPA. In total there have been taken 109 samples (4 duplicate samples of cheese whey, 55 duplicate samples from the reactor of hydrogen production and 50 duplicate samples from the reactor of methane production). In the toxicity tests applied (Thamnotoxkit F, Daphtoxkit FTM magna and zebrafish), L(E)C50 in 24h and 48h were calculated according to the tests protocols. It was found that mean LC50 values for the Thamnocephalus platyurus was 0.76 for the samples from H2-CSTR reactor and 1.33 for the samples from CH4-CSTR reactor. The corresponding values for Daphnia magna in the 48 hours was 1.82 and 2.26 while for the zebrafish 0.88 and 0.95 for the same period. The values resulted from the three tests classify the effluents from “very toxic” to “extremely toxic”. From the cross-correlation of physicochemical parameters with L(E)C50 results of Thamnocephalus platyurus, the samples of H2-CSTR reactor have positive correlation with the ammonium, nitrites and nitrates ions (R= 0.368, R=0.442 and R=0.362 respectively) and the samples of CH4-CSTR reactor have positive correlation only with TDS (R=0.860). Zebrafish correlates with phosphates (R= 0.542) and ammonium ions (R=0.562) for the samples of H2-CSTR, while for the samples of CH4-CSTR correlate with phosphates (R=0.963) and nitrites ions (R= 0.960). Daphnia magna does not show significant correlation. The highest values observed was for the samples of H2-CSTR with the ammonium ions (R= 0.316). According to the above results it is proved that Thamnocephalus platyurus and zebrafish are the most sensitive and suitable organisms for the toxicity estimation of the particular effluents. The observation of spinal malformations in a small percentage of zebrafish, led to the detection of heavy metals to all treated samples. Chromium, manganese, zinc and lead were detected, however the role of each one of them in the observed malformations could not be investigated in the frames of this study and further research is required. In ferentially, all the treated samples with both methods were toxic. They are not environmentally safe for direct disposal in the aquatic receiver and further treatment for the removal of the toxic factors as phosphates and the ions of nitrogen is needed. Finally, it must be noted that the present study is the first toxicity evaluation study of the dairy wastewaters after their treatment for biogas not only in the Greek area but worldwide. Since such a treatment constitutes an innovative and combined methodology that aims to the remediation of the wastewaters and the production of a beneficial by-product as biogas, the results of this study could constitute a useful tool, in one hand for the scientists concerning wastewater treatment provoking them to search methods for further remediation of these effluents from the remaining toxic factors and on the other hand for the officials in order to achieve monitoring and controlling of these particular units.

A avaliação das populações de ácaros e colêmbolos edáficos tem recebido atenção com vistas ao uso como indicadores biológicos da qualidade do solo devido às importantes funções desempenhadas por estes no solo. A análise do número, frequência e diversidade destes grupos em áreas sob cultivo agrícola, em comparação com áreas sob vegetações nativas podem indicar alterações ocorridas no solo. Com base nesta hipótese, este trabalho buscou avaliar qualitativa e quantitativamente a população de ácaros e colêmbolos presentes em uma área com o cultivo de Eucalyptus sp, uma área de mata nativa e outra de campo nativo, localizadas na Estação Experimental Agronômica da UFRGS, Eldorado do Sul – RS. Para isso, foram coletadas mensalmente, no período de janeiro a setembro de 2009, amostras de solo de três áreas. As amostras foram coletadas utilizando-se cilindros metálicos com 7,0 cm de diâmetro e 7,5 cm de altura, que eram introduzidos no solo e depois de retirados, eram envoltos em filme plástico de PVC e transportados em caixas de isopor até o laboratório de microbiologia da Faculdade de Agronomia da UFRGS. Para a extração dos ácaros e colêmbolos das amostras utilizou-se o método do funil de Berlese-Tullgren adaptado. Após eram recolhidos em frascos de vidro com líquido preservante (70% álcool e 1% glicerol). Após 168 horas de extração, todos os espécimes foram contados e identificados com auxílio de lupa e microscópio. Durante o período do estudo, nas três áreas foram capturados 1.314 ácaros e colêmbolos. O solo de área sob mata nativa apresentou o maior número de ácaros e colêmbolos em comparação com as demais áreas, mostrando que as diferentes coberturas vegetais podem influenciar na presença de indivíduos. Nas condições deste trabalho, a flutuação populacional de ácaros da família Oribatidae não foi afetada pelas diferentes coberturas vegetais do solo estudado não sendo um bom indicador da qualidade biológica do solo. A flutuação populacional dos ácaros das famílias Eupodidae e Pachygnathidae e dos colêmbolos da família Hypogastruridae podem ser indicados como bioindicadores de qualidade do solo. / The study of populations of mites and springtails edaphic has received attention in order to use as biological indicators of soil quality because of the important functions performed by them in the ground. The analysis of the number, frequency and diversity of these groups in areas under cultivation, compared with areas under native vegetation may indicate changes in the soil. Based on this hypothesis, this study sought to evaluate qualitatively and quantitatively the population of mites and springtails present in an area with growing Eucalyptus spp, a native forest and other native grassland, located at the Agronomic Experimental Station of UFRGS, Eldorado do Sul - RS. To do so, were collected monthly between January to September 2009, soil samples from three areas. Samples were collected using a metal cylinder with diameter 7.0 cm and 7.5 cm in height, which were introduced into the soil and once removed, were wrapped in PVC film and transported in coolers to the laboratory of Microbiology, Faculty of Agronomy. For the extraction of mites and springtails of the samples used the method of the Berlese-Tullgren funnel adapted. After they were collected in glass bottles with liquid preservative (70% alcohol and 1% glycerol). After 168 hours of extraction, all specimens were identified and counted with a magnifying glass and microscope. During the study period, three areas were captured 1314 mites and springtails. The land area under forest had the largest number of mites and springtails in comparison with other areas, showing that the different vegetation cover may influence the presence of individuals. In the present conditions, the population fluctuations of the mite family Oribatidae was not affected by different vegetation covers the soil studied was not a good indicator of soil biological quality. The mite population fluctuation of families and Eupodidae Pachygnathidae and Collembola family Hypogastruridae can be displayed as bioindicators of soil quality.

VIANA, Sidarta Lopes. Avaliação da toxicidade da água residuária da usina de beneficiamento da casca do coco verde em girinos de Leptodactylus vastus (Amphibia, anura, leptodactylidae). 2010. 78 f. Dissertação (Mestrado em ecologia e recursos naturais)- Universidade Federal do Ceará, Fortaleza-CE, 2010. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-05-27T17:35:19Z No. of bitstreams: 1 2010_dis_slviana.pdf: 817917 bytes, checksum: 92c46a8430dd24028846f5b3adfcdb6b (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-05-27T20:41:24Z (GMT) No. of bitstreams: 1 2010_dis_slviana.pdf: 817917 bytes, checksum: 92c46a8430dd24028846f5b3adfcdb6b (MD5) / Made available in DSpace on 2016-05-27T20:41:24Z (GMT). No. of bitstreams: 1 2010_dis_slviana.pdf: 817917 bytes, checksum: 92c46a8430dd24028846f5b3adfcdb6b (MD5) Previous issue date: 2010 / Amphibians have several characteristics that make them particularly sensitive bioindicators for environmental quality. In this study, tadpoles of the anuran species Leptodactylus vastus A. Lutz, 1930 were used in bioassays to evaluate the toxicity of the liquid from the coconut shell (LCCV), a byproduct of the processing of the green coconut shell. It was analyzed acute effects as lethality and chronic effects as changes in the rate of metamorphosis, in the length of the larval period and in the mass of the tadpoles at metamorphosis. It was performed a total of six experiments, each lasting ninety days, using both LCCV extracted from fresh coconut shells still greenish and LCCV extracted from dry coconut shells that had already became yellowish. The first four experiments had seven treatments each, with concentrations of 0% (control), 0,1%, 0,2%, 0,3%, 0,4%, 0,5% and 0,6% of LCCV. The other two experiments had six treatments each, with concentrations of 0% (control); 0,3%; 0,6%; 0,9%; 1,2% and 1,5% of LCCV. Each treatment had three replicates with ten tadpoles each. The tadpoles were collected at Campus do Pici, Fortaleza, Ceará, in different places and days to avoid pseudo-replication in space and time. The LCCV had a toxic effect on the tadpoles of Leptodactylus vastus, causing an increase in mortality and a decrease in size at metamorphosis, in the highest concentrations tested. However, in some intermediate concentrations, the LCCV contributed to an increase in the mass of tadpoles during metamorphosis. Moreover, in the concentrations tested, the LCCV did not affect the ability of surviving tadpoles to complete metamorphosis or the length of the larval period. Tadpoles of L.vastus showed a positive correlation between mass at metamorphosis and length of the larval period, except in the highest concentrations tested. These results indicate that the LCCV can negatively affect the biota of aquatic environments, making necessary to determine safe parameters of its emission to the environment. The tadpoles of Leptodactylus vastus showed a good potential for use in ecotoxicological studies and as bioindicators of disturbance in aquatic environments, as L.vastus is a quite common species that it is not under threat or danger of extinction, and that produce conspicuous ovipositions with many eggs and larvae relatively large and easily manipulated as they do not require aeration or filtration of water or a special diet. / Os anfíbios apresentam algumas características que os tornam bioindicadores particularmente sensíveis da qualidade ambiental. No presente estudo, girinos da espécie de anuro Leptodactylus vastus A. Lutz, 1930 foram utilizados em bioensaios para avaliar a toxicidade do líquido da casca do coco verde (LCCV), subproduto do processo de beneficiamento da casca do coco verde. Foram analisados efeitos agudos como letalidade, e crônicos, como alterações na taxa de metamorfose, na duração da fase larval e na massa dos girinos por ocasião da metamorfose. Foi realizado um total de seis experimentos, cada um com duração de noventa dias, utilizando tanto LCCV extraído de cascas de coco mais frescas e que conservavam a coloração verde quanto LCCV extraído de cascas de coco mais secas cuja coloração tinha se tornado amarelada. Os quatro primeiros experimentos tiveram sete tratamentos cada, com concentrações de 0% (Controle); 0,1%; 0,2%; 0,3%; 0,4%; 0,5%; 0,6% de LCCV. Os outros dois experimentos tiveram seis tratamentos cada, com concentrações de 0% (Controle); 0,3%; 0,6%; 0,9%; 1,2%; 1,5% de LCCV. Cada tratamento teve três réplicas com dez girinos cada. Os girinos foram coletados no Campus do Pici, Fortaleza, Ceará, em locais e dias variados para evitar pseudo-replicação espacial e temporal. O LCCV apresentou um efeito tóxico sobre os girinos de Leptodactylus vastus, com aumento na letalidade e diminuição no tamanho na metamorfose, nas mais altas concentrações testadas. Entretanto, em algumas concentrações intermediárias, o LCCV contribuiu com um aumento na massa dos girinos por ocasião da metamorfose. Por outro lado, o LCCV nas concentrações testadas, não afetou a capacidade dos girinos sobreviventes de completar a metamorfose nem o tempo de duração da fase larval. Os girinos de L.vastus apresentaram uma correlação positiva entre massa por ocasião da metamorfose e duração do período larval, exceto nas mais altas concentrações de LCCV testadas. Os resultados indicam que o LCCV pode afetar negativamente as comunidades bióticas de ambientes aquáticos, se fazendo necessária a determinação de parâmetros seguros de sua emissão para o meio ambiente. Os girinos de Leptodactylus vastus mostraram um bom potencial de utilização em estudos ecotoxicológicos e como bioindicadores de perturbações em ambientes aquáticos, por sua espécie ser bastante comum e não se encontrar sob perigo ou ameaça de extinção, por suas fêmeas produzirem uma postura conspícua com grande número de ovos e larvas relativamente grandes e de fácil manipulação, e pelo fato destas larvas apresentarem fácil manejo em laboratório, não exigindo aeração ou filtração da água nem rações especiais.

A avaliação das populações de ácaros e colêmbolos edáficos tem recebido atenção com vistas ao uso como indicadores biológicos da qualidade do solo devido às importantes funções desempenhadas por estes no solo. A análise do número, frequência e diversidade destes grupos em áreas sob cultivo agrícola, em comparação com áreas sob vegetações nativas podem indicar alterações ocorridas no solo. Com base nesta hipótese, este trabalho buscou avaliar qualitativa e quantitativamente a população de ácaros e colêmbolos presentes em uma área com o cultivo de Eucalyptus sp, uma área de mata nativa e outra de campo nativo, localizadas na Estação Experimental Agronômica da UFRGS, Eldorado do Sul – RS. Para isso, foram coletadas mensalmente, no período de janeiro a setembro de 2009, amostras de solo de três áreas. As amostras foram coletadas utilizando-se cilindros metálicos com 7,0 cm de diâmetro e 7,5 cm de altura, que eram introduzidos no solo e depois de retirados, eram envoltos em filme plástico de PVC e transportados em caixas de isopor até o laboratório de microbiologia da Faculdade de Agronomia da UFRGS. Para a extração dos ácaros e colêmbolos das amostras utilizou-se o método do funil de Berlese-Tullgren adaptado. Após eram recolhidos em frascos de vidro com líquido preservante (70% álcool e 1% glicerol). Após 168 horas de extração, todos os espécimes foram contados e identificados com auxílio de lupa e microscópio. Durante o período do estudo, nas três áreas foram capturados 1.314 ácaros e colêmbolos. O solo de área sob mata nativa apresentou o maior número de ácaros e colêmbolos em comparação com as demais áreas, mostrando que as diferentes coberturas vegetais podem influenciar na presença de indivíduos. Nas condições deste trabalho, a flutuação populacional de ácaros da família Oribatidae não foi afetada pelas diferentes coberturas vegetais do solo estudado não sendo um bom indicador da qualidade biológica do solo. A flutuação populacional dos ácaros das famílias Eupodidae e Pachygnathidae e dos colêmbolos da família Hypogastruridae podem ser indicados como bioindicadores de qualidade do solo. / The study of populations of mites and springtails edaphic has received attention in order to use as biological indicators of soil quality because of the important functions performed by them in the ground. The analysis of the number, frequency and diversity of these groups in areas under cultivation, compared with areas under native vegetation may indicate changes in the soil. Based on this hypothesis, this study sought to evaluate qualitatively and quantitatively the population of mites and springtails present in an area with growing Eucalyptus spp, a native forest and other native grassland, located at the Agronomic Experimental Station of UFRGS, Eldorado do Sul - RS. To do so, were collected monthly between January to September 2009, soil samples from three areas. Samples were collected using a metal cylinder with diameter 7.0 cm and 7.5 cm in height, which were introduced into the soil and once removed, were wrapped in PVC film and transported in coolers to the laboratory of Microbiology, Faculty of Agronomy. For the extraction of mites and springtails of the samples used the method of the Berlese-Tullgren funnel adapted. After they were collected in glass bottles with liquid preservative (70% alcohol and 1% glycerol). After 168 hours of extraction, all specimens were identified and counted with a magnifying glass and microscope. During the study period, three areas were captured 1314 mites and springtails. The land area under forest had the largest number of mites and springtails in comparison with other areas, showing that the different vegetation cover may influence the presence of individuals. In the present conditions, the population fluctuations of the mite family Oribatidae was not affected by different vegetation covers the soil studied was not a good indicator of soil biological quality. The mite population fluctuation of families and Eupodidae Pachygnathidae and Collembola family Hypogastruridae can be displayed as bioindicators of soil quality.

La Directive Cadre Européenne sur l'eau impose d'évaluer la qualité des cours d'eau au moyen d'indicateurs chimiques et biologiques dont les diatomées font partie. Les indices basés sur la composition taxonomique et l'abondance relative des taxa de diatomées sont robustes. Cependant, de nombreux échantillons doivent être analysés chaque année alors que l'identification de ces micro-algues en microscopie optique est difficile à cause des incertitudes taxonomiques, et nécessite temps et expertise. Ainsi, des améliorations peuvent encore être apportées pour faciliter le suivi en routine de la qualité de l'eau. Les techniques de biologie moléculaire sont des outils efficaces pour identifier les microorganismes et pourraient donc être utilisées pour améliorer l'identification des diatomées. Les objectifs de cette thèse étaient donc de compléter les connaissances sur la taxonomie des diatomées d'eau douce par des méthodes moléculaires et de progresser dans le développement d'un outil moléculaire permettant l'identification des diatomées dans des échantillons naturels, en vue de son utilisation en bioindication. L'étude de la taxonomie de plusieurs groupes de diatomées a été réalisée en combinant des approches morphologiques et des approches moléculaires. Nos travaux ont montré les capacités des séquences ADN pour discriminer les taxa de diatomées et révéler leurs relations phylogénétiques. L'utilisation de séquences ADN a montré que les critères morphologiques utilisés pour identifier les diatomées ne correspondaient pas systématiquement à leurs relations phylogénétiques. L'utilisation de différents marqueurs a permis des discriminations à différents niveaux taxonomiques. Nos résultats ont également révélé l'importance de combiner des approches complémentaires, morphologiques et moléculaires, pour améliorer notre compréhension des relations entre les différents taxa de diatomées et ainsi stabiliser leur taxonomie. Les séquences ADN permettant une discrimination des taxa de diatomées, nous avons testé un outil moléculaire de séquençage haut-débit, le pyroséquençage 454, dans le but d'identifier les taxa composant les communautés de diatomées. Nous avons ainsi assemblé des bases de séquences de référence bénéficiant d'une identification taxonomique. Nous avons également participé au développement d'outils bioinformatiques nécessaires à l'analyse des données de pyroséquençage. Enfin, nous avons pu tester ces outils pour établir des inventaires taxonomiques de diatomées dans des communautés artificielles (mélanges de souches) et dans des communautés environnementales (biofilms d'eau douce). Ces études ont prouvé le potentiel du pyroséquençage 454 pour étudier les communautés de diatomées à des niveaux taxonomiques précis. La comparaison de différents marqueurs nucléiques a révélé que le marqueur rbcL était le marqueur le plus adapté à l'identification des diatomées par pyroséquençage. En effet, en prenant en compte les bases de séquences de référence, la reproductibilité et les biais de la méthode ainsi que le pouvoir résolutif du marqueur, l'utilisation du rbcL a permis la meilleure estimation de la composition en diatomées d'échantillons complexes. Des progrès devront encore être faits avant de pouvoir utiliser les outils moléculaires pour évaluer la qualité de l'eau par les diatomées. Cependant nos différentes études permettront de guider les prochaines analyses de manière à aboutir à un suivi de la qualité de l'eau basé sur des inventaires moléculaires des taxa de diatomées. / The European Water Framework Directive requires assessing the river quality using chemical and biological indicators among which are diatoms. Indices based on taxonomic composition and relative abundance of diatom taxa are robust. However, thousands of diatom samples are analyzed every year while microscopic identification is difficult due to taxonomic uncertainties, and requires time and expertise. Thus, it is still possible to improve the routine monitoring of water quality. The molecular biology techniques are accurate tools to identify microorganisms and could be used to enhance diatom identification. The objectives of this thesis were therefore to improve our knowledge on the freshwater diatom taxonomy by molecular methods and to progress in the development of a molecular tool in order to identify diatoms in natural samples, thus improving bioindication tools. The taxonomic study of several groups of diatoms was achieved by combining morphological and molecular approaches. Our results showed the capacity of DNA sequences to discriminate diatom taxa and revealed their phylogenetic relationships. The use of DNA sequences showed that the morphological criteria used to identify diatoms do not correspond systematically to their phylogenetic relationships. The use of different markers allowed discrimination at different taxonomic levels. Our results also revealed the importance of combining complementary approaches, morphological and molecular, to improve our understanding of the relationships between different diatom taxa and thus stabilize their taxonomy. As DNA sequences allowed discrimination of diatom taxa, we tested a molecular tool of high throughput sequencing, 454 pyrosequencing, in order to identify the diatom composition of communities. We assembled reference libraries of sequences linked to taxonomic identification and we contributed to the development of bioinformatic tools required to analyze data from pyrosequencing. Finally, we tested these tools to establish taxonomic inventories of diatoms in artificial communities (mixtures of strains) and environmental communities (freshwater biofilm samples). The studies showed the potential of 454 pyrosequencing to accurately analyze diatom communities. The comparison of different nucleic markers revealed that the rbcL marker was the most suitable to identify diatoms using pyrosequencing. Indeed, taking into account reference libraries, reproducibility and bias of the method, and the resolving power of marker, the use of rbcL allowed the best estimation of the diatom composition in complex samples. Improvements will be necessary to use molecular tools in order to assess water quality using diatoms. However our studies lead the way for future experiments in order to achieve a monitoring of water quality based on molecular inventories of diatom taxa.