Petroleum hydrocarbons are the most widespread contaminants in the environment. Interest in the biodegradation of polycyclic aromatic hydrocarbons and compounds (PAHs/PACs) is motivated by their ubiquitous distribution, their low bioavailability, high persistence in soils and their potentially deleterious effects to human health. Identifying the diversity of microorganisms that degrade PAHs/PACs can be utilised in the development of bioremediation techniques. Understanding the mechanisms of bacterial populations to adapt to the presence of pollutants and the extent that lateral transfer of key functional genes occurs, will allow the exploitation of microbial PAC/PAH-degradative capabilities and therefore enhance the successful application of bioremediation strategies. A key aim of this study was to isolate and identify PAC-degrading bacteria for potential use in future bioremediation programmes. A series of PAC enrichments were established under the same experimental conditions from a single sediment sample taken from a highly polluted estuarine site. Distinct microbial community shifts were directly attributable to enrichment with different PAC substrates. The findings of this study demonstrate that five divisions of the Proteobacteria and Actinobacteria can degrade PACs. By determining the precise identity of the PAC-degrading bacteria isolated, and by comparing these with previously published research, this study showed how bacteria with similar PAC degrading capabilities and 16S rRNA signatures are found in similarly polluted environments in geographically very distant locations e.g. China, Italy, Japan and Hawaii. Such a finding suggests that geographical barriers do not limit the distribution of key PAC-degrading bacteria. This is significant when considering the diversity and global distribution of microbes with PAC-degradative capabilities and the potential for utilising these microbial populations in future bioremediation strategies. In the laboratory, enrichment of bacteria able to utilise PAHs has commonly been performed in liquid media, with the PAH dissolved in a carrier solvent. This study found the presence of a carrier solvent significantly affects the resultant microbial population. Although the same sediment sample was used as the bacterial source in all enrichments, different bacterial strains were obtained depending upon the presence of the carrier solvent and the PAH. This is important when considering appropriate methodology for the isolation of PAH-degrading bacteria for future bioremediation programmes. Additionally, the species comprising the resultant population of the enrichment when a carrier solvent was present were similar to previously reported PAH-degrading species. Such a finding necessitates review of previously reported PAH-degrading bacterial species that have been isolated and identified from enrichments using a carrier solvent. Understanding how bacteria acclimatise to environmental pollutants is vital for exploiting these mechanisms within clear up strategies of contaminated sites. Two major lineages of the α subunit of PAH dioxygenases were identified: Actinobacteria and Proteobacteria. Comparison of the α subunit phylogeny with the 16S rRNA phylogeny implies that the PAH-dioxygenases evolved prior to the separation of these phyla or that lateral transfer occurred in the very distant past. No evidence for lateral transfer of the α subunit between the Actinobacteria and Proteobacteria was found in the phylogenetic analyses of this research. Multiple lateral transfer events were inferred between the species of the Actinobacteria and between the classes of the Proteobacteria. The clustering of the taxa within the α subunit phylogeny indicates that lateral transfer of the α subunit gene occurred after the separation of the classes of Proteobacteria and also after the speciation of the γ-Proteobacteria. These findings reveal how bacteria have acclimatised to PAH pollutants through multiple lateral transfer events of a key PAH-degradative gene. This knowledge of the transfer of genetic material will broaden our prospects of exploiting microbial populations.

Η παρούσα μελέτη διερευνά την ικανότητα των πολυκυκλικών αρωματικών υδρογονανθράκων να (PAHs), όπως είναι το φαινανθρένιο και το ανθρακένιο, να επάγουν συνθήκες stress (stress syndrome) σε ιστούς (π.χ. βράγχια) του είδους Mytilus galloprovincialis. Σύμφωνα με τα αποτελέσματα αυτής της μελέτης, τα μύδια τα οποία εκτέθηκαν για 7 ημέρες σε PH και/ή AN (τελικής συγκέντρωσης 0,1 mg/L σε κάθε περίπτωση) καθώς και σε μίγμα αυτών των οργανικών ρύπων (τελικής συγκέντρωσης 0,2 mg/L), παρουσίασαν αυξημένη συχνότητα εμφάνισης πυρηνικών ανωμαλιών, όπως είναι οι μικροπυρήνων (MN), και επαγωγή νευρολογικών διαταραχών, πράγμα που υποδεικνύεται από την μέτρηση της δραστικότητας της ακετυλχολινεστεράσης (AChE). Επιπλέον, τα αυξημένα επίπεδα της MDA στα βράγχια των εκτιθέμενων ατόμων καθώς και η μείωση του χρόνου ημιζωής (LT50) των μυδιών που εκτέθηκαν στον αέρα (τεχνική Stress on Stress), αντανακλούν στις οξειδωτικές επιπτώσεις που επάγουν οι πολυκυκλικοί αρωματικοί υδρογονανθράκες στους ιστούς των εκτιθέμενων στον αέρα μυδιών, πράγμα που οδηγεί στην επαύξηση των τοξικών επιδράσεων, σε διαφορετικά επίπεδα της λειτουργίας του οργανισμού. / This study investigates the ability of certain polycyclic aromatic hydrocarbons (PAHs), such as phenanthrene (PH) and anthracene (AN), to induce stress conditions (stress syndrome) in tissues (e.g. gills) of mussel Mytilus galloprovincialis. According to the results of this study, mussels exposed for 7 days to PH and/or AN (at a final concentration of 0,1mg/L in each case) as well as to a mixture of these organic pollutants (at a final concentration of 0,2 mg/L) showed increased frequencies of nuclear abnormalities, such as the presence of micronuclei (MN), and induction of neurological disorders, as shown by measuring the activity of acetylcholinesterase (AChE). Furthermore, increased levels of MDA in the gills of exposed individuals, and the reduction of the lethal time (LT50) of challenge mussels superimposed in air (Stress on Stress technique), merely reflect PAHs-induced oxidative effects on tissues of challenge mussels, thus leading to the enhancement of toxic effects to different levels of organism function.

No description available.

Soils and groundwaters face an influx of contaminants, many of which are known to be hazardous to the public's health. Several remediation technologies have been developed to clean-up contaminated soils, but additional information on the behavior of organic chemicals in the subsurface environment is needed for effective remediation. The purpose of this study was to investigate the relationship between sorption and biodegradation on the ultimate fate of subsurface contaminants. Sorption and biodegradation of phenanthrene, a hydrophobic polycyclic aromatic hydrocarbon, were modeled using soil and water microcosms. The soils were prepared in the laboratory from a parent soil to obtain sorbents that were similar in all respects except for soil organic matter content. Biodegradation was evaluated by production of CO2 and disappearance of phenanthrene from the sorbed and aqueous phases. The sorption of phenanthrene to each soil occurred in two distinct stages. The majority of phenanthrene mass sorbed during the first few days. After the initial rapid sorption period, the sorption rate continually decreased. The time required for complete sorption varied according to soil organic matter content, with soils having higher organic matter contents requiring more time for complete adsorption. Desorption of phenanthrene from soils was hysteretic, although it appeared that phenanthrene would completely desorb. Phenanthrene desorption rates decreased as the chemical exposure time increased. The biodegradation of phenanthrene also occurred with a fast and slow phase. The slowly degradable fraction increased with soil organic matter content. The data suggest that the biodegradation of phenanthrene is desorption controlled. / Master of Science

L'ingénierie écologique a permis l'émergence de nouvelles technologies telles que la phytoremédiation, une approche de dépollution des sols basée sur l'utilisation des plantes. Cependant, jusqu'à présent, les mécanismes physiologiques et moléculaires qui contrôlent la dégradation des polluants organiques, en particulier les HAPs, restent très mal documentés chez les plantes supérieures. Dans ce travail, une approche intégrative et un profilage trancriptomique à l'échelle du génome entier ont permis d'étudier les premiers événements moléculaires impliqués dans la réponse, chez les plantes, à un stress provoqué par le phénanthrène. Un réseau de gènes susceptibles d'intervenir dans la perception, la dégradation et/ou la transformation des HAPs en molécules moins dangereuses a ainsi été identifié. D'autre part, nous avons pu montrer que le saccharose induit une tolérance à ce polluant, qui se traduit (i) par une activité transcriptionnelle très rapide, ce qui suggère un effet signalétique, (ii) une reconfiguration importante à l'échelle transcriptionnelle du génome exprimé, qui conduit à une mise en place des voies métaboliques de production d'énergie, de détoxification et de réparation cellulaire. Parallèlement, un site pilote de dépollution a été conçu et mis en œuvre, en exploitant les données obtenues au laboratoire. Cette expérimentation, sur deux ans, nous a permis de comparer l'efficacité de plusieurs espèces végétales mais aussi de tester l'effet protecteur du saccharose in-situ. Malgré l'obtention de résultats contrastés, cette étude confirme l'incidence de la nature des espèces végétales et des pratiques culturales sur l'efficacité de cette méthode de dépollution. Elle souligne également le rôle important de l'interaction plante-microorganismes du sol. / Ecological engineering strategies allowed the emergence of new technologies such as phytoremediation to clean up environmental pollution. Polycyclic Aromatic Hydrocarbons (PAHs) are ubiquitous and hazardous molecules for natural ecosystems and human health. However, molecular mechanisms involved in PAHs detoxification in plant remain largely unknown. This work allowed to decipher the early plant response of phenanthrene induced stress through an integrative investigation using wide genome transcriptomic and targeted metabolomic approaches. Hence, a gene network was identified to be involved in the early perception and signalization of PAHs and putative degradation and/or transformation to harmless molecules. Additionally, we showed that sucrose mitigate phenanthrene induced stress, while Arabidopsis development was arrested and seedlings were unable to accumulate chlorophyll in the presence of high phenanthrene concentration, sucrose allowed growth and chlorophyll accumulation. These phenotypical changes were associated with (i) very early transcriptional regulation, suggesting a sucrose signaling effect (ii) reconfiguration of wide genome transcriptional changes, as metabolic, detoxification and cell repair pathways were induced. In parallel, based on previous results accumulated in controlled conditions. Field investigation allowed the evaluation of the ability of different plant species to clean up PAHs polluted soil. We also evaluate the impact of sucrose supply on natural phytoremediation efficiency. Despite contrasted results, this study confirms the incidence of the plant species and the cultural practices on the efficiency of this method. It also underlines the role of soil microorganisms and plant interactions.

Ο στόχος την παρούσας εργασίας είναι η αποδόμηση των PAHs με χρήση όζοντος.Για το σκοπό αυτό επιλέξαμε το φαινανθρένιο ως ένωση-μοντέλο σε οργανικό διαλύτη, με απώτερο στόχο την επεξεργασία επικίνδυνων αποβλήτων. / The perpose of this study is the degradation of polycyclic aromatic hydrocarbons with ozon. For this reason we use phenanthrene like model-compound in organic solution.

Μέχρι σήμερα η επίδραση της αλατότητας στη διεργασία της ρόφησης των οργανικών ρύπων σε στερεούς ροφητές έχει μελετηθεί θεωρώντας ότι η ρόφηση είναι μία γραμμική διεργασία που εξαρτάται από το κλάσμα του οργανικού άνθρακα. Μελέτες της τελευταίας δεκαετίας έχουν αποδείξει ότι η ρόφηση των οργανικών στο περιβάλλον δεν είναι απαραίτητα μία γραμμική διεργασία. Για ιζήματα και εδάφη με σωματιδιακούς ανθρακούχους ρύπους, όπως είναι τα σωματίδια της αιθάλης ή των εξανθρακωμάτων, έχουν παρατηρηθεί ισόθερμες καμπύλες οργανικών ρύπων με μεγάλη έλλειψη γραμμικότητας. Προκαταρκτικές μελέτες έχουν δείξει ότι πράγματι η αλατότητα επηρεάζει τη ρόφηση ανάλογα με το μηχανισμό που παρατηρείται για κάθε είδος ροφητή. Αλλιώς επηρεάζεται η ρόφηση από την αλατότητα, όταν λαμβάνει χώρα στην επιφάνεια του ροφητή, και αλλιώς στους πόρους στο εσωτερικό του ροφητή. Η παρούσα μεταπτυχιακή εργασία περιλαμβάνει πειραματική μελέτη της ισορροπίας της ρόφησης ενός οργανικού ρύπου (του φαινανθρενίου) σε διάφορους ροφητές από το γλυκό και το θαλασσινό νερό. Οι ροφητές περιλαμβάνουν ανθρακούχα σωματίδια και ετερογενή υλικά, όπως γαιάνθρακα, λιγνίτη και θαλάσσια ιζήματα από την περιοχή του Αλιβερίου, του Λαυρίου και του Σαρωνικού κόλπου (Λουτρόπυργος) που έχουν ρυπανθεί από τα παραπάνω ανθρακούχα σωματίδια. Η αλατότητα φαίνεται να επηρεάζει σημαντικά τη ρόφηση του φαινανθρενίου σε σωματίδια λιθάνθρακα όταν τα πειράματα διεξάγονται σε σχετικά χαμηλές συγκεντρώσεις (<200 μg/L). / Until recently sorption of organic compounds onto sediments was believed to be a linear process dependent on the organic carbon fraction content. Non linear sorption and heterogeneous behavior based has been recognized the last decade. Due to salinity, various effects are observed for the different carbonaceous materials on the capacity and also on the degree of nonlinearity. Each material presents a different sorption pattern and various effects due to salinity are observed (e.g. coal sorption isotherm becomes linear in the presence of salinity whereas for lignite no significant difference is observed). In the present work, we study the sorption behavior of phenanthrene onto different sorbents in salt water. Sorbents used include carbonaceous materials, coal, lignite, and marine sediments from Aliveri, Laurio, and Saronikos gulf that are polluted with carbonaceous particles. Salinity seems to affect significantly the sorption of phenanthrene onto coal particles if the experiments are performed at concentrations lower than 200 μg/L.

Made available in DSpace on 2015-03-26T13:53:05Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1808310 bytes, checksum: ca499fceff48952d8454c181384108c7 (MD5) Previous issue date: 2008-07-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Petroleum is the main source of fuel, and the operations of exploration, transportation, refining and distribution are potential sources of environmental pollution. In contaminated environments, the degradation of PAHs (Polycyclic Aromatic Hydrocarbons) depends on climatic factors, soil and microbial population. This study aimed to assess the potential for degradation of naphthalene, phenanthrene and benzo (a) pyrene hydrocarbons in different soils and sediments from coastal regions, both tropical and polar (Antarctica). The samples were collected in three regions, and (1) main island in the archipelago of Fernando de Noronha, (2) in the Prado coast, in southern Bahia, and (3) Islands in the South Shetlands archipelago, Antarctica. After obtaining samples of TFSA (Earth Thin Air Dried), the soils were submitted to physical and chemical analysis, and experiments were set to evaluate the degradation through respirometry, and tested the PAHs naphthalene, phenanthrene and benzo (a) pyrene. Most degradation of naphthalene was observed in soil contaminated by oil derived from oil, in Fildes (Antarctica), which was attributed to the selection of microbial populations in the efficient use of HC. The rates of degradation of phenanthrene and benzo (a) pyrene were lesser in comparison to naphthalene, which have lower molecular weight and greater solubility. The results of this study suggest that the microorganisms present in these soils, wherever there was no impact of oil spill, were not able to efficiently degrade the oil naphthalene, phenanthrene and benzo (a) pyrene, in experimental conditions tested. That would, after selection, the use of these microorganisms (adapted), the remediation of contaminants soluble in other similar areas of Antarctica. The differences between the rates of degradation of hidrocarbons relate to the levels of TOC (Total Organic Carbon) and TN (Total Nitrogen) soil of local noncontaminated. / O petróleo representa a principal fonte de combustível da humanidade, e as operações de exploração, transporte, refino e distribuição representam fontes potenciais de poluição ambiental. Em ambientes contaminados, a degradação dos HPAs (Hidrocarbonetos Policíclicos Aromáticos) depende de fatores climáticos, dos tipos de solo e das populações microbianas presentes. O presente trabalho teve por objetivo avaliar o potencial de degradação dos hidrocarbonetos naftaleno, fenantreno e benzo(a)pireno em diferentes solos e sedimentos provenientes de regiões costeiras, oceânicas tropicais e polares (Antárticas). As amostras foram coletadas em três regiões, sendo (1) na ilha principal do Arquipélago de Fernando de Noronha, (2) no litoral do Prado, no sul da Bahia, e (3) nas Ilhas do Arquipélago Shetlands do Sul, Antártica. Após a obtenção das amostras de TFSA (Terra Fina Seca ao Ar), os solos foram submetidos a análises químicas e físicas, e foram montados experimentos de degradação através de respirometria, sendo testados os HPAs naftaleno, fenantreno e benzo(a)pireno. A maior degradação de naftaleno foi observada em solo contaminado por hidrocarbonetos derivados de petróleo, em Fildes (Antártica), a qual foi atribuída à seleção de populações microbianas eficientes na utilização desse HC. A maior degradação do fenantreno e do benzo(a)pireno nos solos do Brasil ocorreu na area P5 (Manguezal de Cumuruxatiba), sendo neste caso atribuído à maior disponibilidade de nutrientes. As taxas de degradação de fenantreno e benzo(a)pireno foram menores em comparação do que a do naftaleno, que possui menor peso molecular e maior solubilidade. Os resultados deste trabalho demostram que os microrganismos presentes nos solos em estudo, onde não houve impacto conhecido de derramamento de óleo, não foram capazes de degradar eficientemente os hidrocarbonetos naftaleno, fenantreno e benzo(a)pireno, nas condições experimentais testadas. Nestes ambientes, as diferenças entre as taxas de degradação dos hidrocarbonetos se relacionam com os teores de COT (Carbono Orgânico Total) e NT (Nitrogênio Total).

Dissertação(mestrado) - Universidade Federal do Rio Grande, Programa de Pós–Graduação em Oceanografia Biológica, Instituto de Oceanografia, 2012. / Submitted by Cristiane Gomides (cristiane_gomides@hotmail.com) on 2013-11-18T15:23:39Z No. of bitstreams: 1 anderson.pdf: 2147843 bytes, checksum: 1fd27dd610099252c3f5f6c1e2b14fed (MD5) / Approved for entry into archive by Angelica Miranda (angelicacdm@gmail.com) on 2013-11-18T19:21:19Z (GMT) No. of bitstreams: 1 anderson.pdf: 2147843 bytes, checksum: 1fd27dd610099252c3f5f6c1e2b14fed (MD5) / Made available in DSpace on 2013-11-18T19:21:19Z (GMT). No. of bitstreams: 1 anderson.pdf: 2147843 bytes, checksum: 1fd27dd610099252c3f5f6c1e2b14fed (MD5) Previous issue date: 2012 / A poluição aquática em zonas costeiras é crescente, exigindo cada vez mais o uso de ferramentas alternativas para seu monitoramento. Dentre estas, as de maior potencial são os biomarcadores, que são medidas biológicas capazes de expressar exposição ou efeito deletério de contaminantes. Porém, a falta de informação sobre respostas de biomarcadores a contaminantes em concentrações ambientalmente relevantes constitui empecilho ao emprego destas ferramentas. Neste trabalho foram analisados os efeitos de três contaminantes em diversos biomarcadores usualmente utilizados em estudos de avaliação ambiental. Os contaminantes testados foram cobre (metal), atrazina (herbicida) e fenantreno (hidrocarboneto policíclico aromático). Os biomarcadores avaliados foram atividades enzimáticas (superóxido dismutase, catalase, glutationa redutase e glutationa s-transferase), concentração de metalotioneínas, espécies reativas de oxigênio, capacidade antioxidante, peroxidação lipídica, quebras na fita de DNA, freqüência de células micronucleadas e de anomalias nucleares, e composição iônica corporal. Os três contaminantes demonstraram potencial para afetar os biomarcadores avaliados, porém cada um afetou distintamente os grupos de biomarcadores. O efeito do cobre foi caracterizado por um elevado potencial de geração de estresse oxidativo em brânquias, fígado e músculo, além de causar dano genético através da quebra da fita de DNA e clastogenicidade. A atrazina aumentou a atividade da glutationa s-transferase, causou estresse oxidativo branquial e danos genéticos não detectáveis no ensaio cometa. Por fim, o fenantreno gerou estresse oxidativo muscular, sendo a maioria dos danos possivelmente relacionada à geração de metabólitos reativos durante sua degradação, resultando em elevada genotoxicidade e citotoxicidade, bem como perturbações de sistemas enzimáticos e da fisiologia do animal. / Aquatic pollution in coastal areas is increasing, demanding the use of alternative tools for its adequate monitoring. Among them, biomarkers are those showing higher potential for practical use. They are biological measurements capable of detecting the exposure and effects of contaminants. However, the lack of information on the response of biomarkers to contaminants at environmentally relevant levels is a constraint for the employment of these tools in monitoring programs. In the present study, the effects of three contaminants on several biomarkers commonly employed in environmental analyses were evaluated. Contaminants tested were copper (metal), atrazine (herbicide) and phenanthrene (polycyclic aromatic hydrocarbon). Biomarkers evaluated were enzyme activities (superoxide dismutase, catalase, glutathione reductase and glutathione s-transferase), metallothioneins concentration, reactive oxygen species, antioxidant capacity, lipid peroxidation, DNA strand-breaks, micronucleated cells and nuclear anomalies frequencies, and whole-body ion composition. The three contaminants showed potential to affect the biomarkers evaluated. However, each contaminant affected distinctly the groups of biomarkers. Copper effect was characterized by a elevated potential for oxidative stress generation in gills, liver and muscle. It also induced genetic damage (DNA strand-breaks) and clastogenicity. Atrazine increased glutathione stransferase activity, caused oxidative stress in gills and genetic damage non detectable by the Comet assay. Finally, phenanthrene induced oxidative stress in muscle, being most of the damage likely associated with the generation of reactive metabolites during the contaminant degradation process. Exposure to phenanthrene resulted in elevated genotoxicity and cytotoxicity, as well as perturbations of the enzymatic systems and the whole-animal physiology.

The gas-phase ion chemistry of hydroxyl- and amino-substituted polycyclic aromatic hydrocarbons (PAHs) and their protonated counterparts were studied using mass spectrometry. Ions were generated using an electron ionization (EI) source and the unimolecular chemistry of metastable ions was studied by performing mass-analysed ion kinetic energy spectrometry (MIKES) experiments with a magnetic sector tandem mass spectrometer. Collision-induced dissociation (CID) experiments were used in conjunction with MIKES experiments to determine ion structure. The ten molecules studied were: 1-naphthol, 2-naphthol, 1-naphthylamine, 2-naphthylamine, 1-aminoanthracene, 2-aminoanthracene, 1-phenanthrol, 9-phenanthrol, 1-hydroxypyrene and 1-aminopyrene. Since it is believed that larger PAHs, on the order of more than 50 carbon atoms, populate the interstellar medium, the goal of this study was to attempt to extrapolate the results from smaller systems to larger ones. The trends found include: hydroxy-substituted PAH radical cations lose carbon monoxide spontaneously and amino-substituted PAH radical cations lose HCN. Mechanisms for both processes are proposed, and it appears from the present results that this process should extrapolate to larger PAHs. Another trend found was that all the remaining fragment ions were always a closed ring. Protonated amino-substituted PAHs were generated by electrospray ionization using a quadruple time-of-flight mass spectrometer. By protonating 1-naphthol and 2-naphthol using methane in the high-pressure EI source, it was found that they lost exclusively H2O. As for 2-naphthylamine, 1-aminoanthracene and 2-aminoanthracene, it was found that 2-naphthylamine lost NH3 and a hydrogen atom, NH3being the dominant channel. However, as the ion size 3 increases, the hydrogen-loss channel became the dominant channel. This means that larger PAHs will likely lose exclusively a hydrogen atom to reform the parent radical cation.