The chemical characterization of sediments accumulated in catch basins and stormwater ponds provides important information for assessing risks associated with management of these residuals upon removal of accumulated deposits in stormwater systems. In this study, over a period of 15 months, more than 150 residual samples were collected from 77 catch basin units and 22 stormwater ponds from 16 municipalities throughout the state of Florida. Concentrations (mg/kg) of metals and metalloids (arsenic, barium, cadmium, chromium, copper, lead, mercury, nickel, selenium, silver, and zinc) and trace organics (volatile organics, semi-volatile organics, herbicides, and pesticides) in the sediments were measured. In addition, the synthetic precipitation leaching procedure (SPLP) was utilized to evaluate pollutant leachability risk for a subset of the samples collected. Measured pollutant concentrations were compared to corresponding risk-based guidelines in Florida (i.e., Florida soil cleanup target levels) to assess potential human health risks of beneficial use of these residuals through land application. Leached concentrations were compared to risk-based water quality guidelines (i.e., Florida groundwater cleanup target levels) to examine the potential for groundwater contamination. Although several metals (arsenic, barium, chromium, copper, nickel, lead, and zinc) were routinely detected in the catch basin and stormwater pond sediments, their concentrations were generally lower than the Florida's risk-based cleanup target levels for soils. A small number of organochlorine compounds (e.g., 4,4′-DDE, 4,4′-DDT) were detected, but only in a limited number of the samples (less than 10%); leaching of trace organic pollutants above the Florida risk-based groundwater thresholds was rare. The results suggest that when land-applied or beneficially used, these residuals are not expected to pose a significant threat to human health or the environment and the results of this research will provide stormwater managers and environmental management authorities with a useful resource to examine proper disposal and beneficial use of catch basin and stormwater pond sediments.

Disposal and beneficial-use options for street sweeping residuals collected as part of routine roadway maintenance activities in Florida, USA, were assessed by characterizing approximately 200 samples collected from 20 municipalities. Total concentrations (mg/kg or μg/kg) and leachable concentrations (mg/L or μg/L) of 11 metals and a number of organic pollutant groups (volatile organics, semi-volatile organics, pesticides, herbicides, carbamates) in the samples were measured. The synthetic precipitation leaching procedure (SPLP) was performed to evaluate the leachability of the pollutants. From the total metal analysis, several metals (e.g., arsenic, barium, chromium, copper, nickel, lead, and zinc) were commonly found above their detection limits. Zinc was found to have the highest mean concentration of all metals measured (46.7 mg/kg), followed by copper (10.7 mg/kg) and barium (10.5 mg/kg). The metal with the smallest mean concentration was arsenic (0.48 mg/kg). A small fraction of the total arsenic, barium, lead, and zinc leached in some samples using the SPLP; leached concentrations were relatively low. A few organic compounds (e.g., 4,4′-DDT, endrin, and endosulfan II) were detected in a limited number of samples. When the total and leaching results were compared to risk-based Florida soil cleanup target levels and groundwater cleanup target levels, the street sweepings were not found to pose a significant human-health risk via direct exposure or groundwater contamination.

Bulk atmospheric deposition samples were taken every two months for a year in 2009-2010 at two sites in northern Sweden and analyzed for a suite of legacy and emerging persistent organic chemicals including legacy organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs)  and novel flame retardants (NFRs).  To further investigate the urban occurrence and contribution to remote contamination of flame retardants, indoor air, ventilation system air, and dust were sampled in several microenvironments in Stockholm during the winter of 2012 for analysis of PBDEs, isomer-specific hexabromocyclododecane (HBCDD), and NFRs.  Outdoor air and soil samples were also sampled around the same time period for analysis of the same compounds.  Five emerging pollutants were detected in atmospheric deposition: the current-use pesticides trifluralin and chlorothalonil; and the NFRs 1,2-dibromo-4-(1,2-dibromoethyl)cyclohexane (TBECH), 1,2-bis(tribromophenoxy)ethane (BTBPE), and Dechlorane Plus (DP).  A decrease in the fraction of the anti isomer of DP was observed at the more remote site, indicating isomer-selective degradation or isomerization during long range atmospheric transport.  The more remote site also received more total deposition of organic pollutants despite its receiving less precipitation.  Although PBDEs and emerging organic pollutants were detected, the bulk of the deposition consisted of PCBs and hexachlorocyclohexanes (HCHs) at both sites.   Several NFRs were identified in indoor and ventilation air samples including TBECH, pentabromotoluene (PBT), hexabromobenzene (HBB), 1,2-bis(pentabromodiphenyl)ethane (DBDPE), 2-ethylhexyl-2,3,4,5-tetrabromobenzoate (EHTBB), and bis(2-ethylhexyl)-3,4,5,6-tetrabromo-phthalate (TBPH).  There was no significant difference between concentrations of contaminants in ventilation system air and indoor air indicating that the flame retardants included in this study reach the outdoor environment via ventilation systems.  Dust and outdoor air samples have only been analyzed for HBCDDs so far.  HBCDD concentrations in apartments, offices, and schools were lower than reported concentrations for other countries in similar microenvironments.   However, an enrichment of α-HBCDD compared to γ-HBCDD was observed in dust taken from near treated products in stores when compared to dust taken directly from those products.  Very low concentrations of HBCDDs were detected in outdoor air.

<p>U doktorskoj disertaciji evaluirani su rezultati kvantifikovanih koncentracionih nivoa lipofilnih perzistentnih i emergentnih organskih polutanata u uzorcima sedimenta kolektovanih sa deset reprezentativnih lokaliteta u srednjem Podunavlju. Na osnovu sprovedenih laboratorijskih i terenskih istraživanja definisan je prostorni trend jedinjenja na ispitivanom području. Primenom multivarijantnih tehnika eksperimentalni rezultati su uspešno modelovani statističkim metodama koje su izdiferencirale izvore kontaminacije za ukupan set ispitivanih polutanata. Prikazana je procena uticaja kontaminiranog sedimenta na akvatične organizme i humanu populaciju prema stepenu kancerogenosti jedinjenja. Predstavljeni su različiti scenariji izloženosti ingestijom i dermalnim kontaktom, u zavisnosti od vremena ekspozicije i izložene površine potencijalnih receptora, različitog uzrasta i pola. Po prvi put u istraživanom području, implementirana je ex-situ ravnotežna metodologija pasivnog uzorkovanja primenom sorpcionog medijuma od silikonske gume testirane pri različitim masenim odnosima polimer-uzorak sedimenta. Primenjenom metodom uspešno su dobijene slobodno rastvorene koncentracije lipofilnh kontaminanata u pornoj vodi sedimenta, kao prediktora za određivanje biodostupnosti jedinjenja.</p> / <p>The doctoral dissertation evaluated the results of quantified concentration levels of lipophilic persistent and emergent organic pollutants in sediment samples collected from ten representative localities in the central Danube region. Based on the conducted laboratory and field research, the spatial distribution of compounds in the studied area was defined. Using multivariate techniques, the experimental results were successfully modeled by statistical methods that differentiated the sources of contamination for the total set of tested pollutants. A risk assessment of contaminated sediment on aquatic organisms and the human population according to carcinogenicity of the tested compounds is presented in thesis. Different exposure scenarios of ingestion and dermal contact, depending on the time exposure and exposed surface skin of potential receptors, of different ages and sex, are presented. For the first time in the study area, an ex-situ equilibrium passive sampling methodology was implemented using silicone rubber sorption medium tested at different polymer-sample sediment mass ratios. Using the method, the freely dissolved concentrations of lipophilic contaminants in pore water were successfully obtained as predictors for determining the bioavailability of the compounds.</p>

Successful bioremediation of polluted soils is often limited by the bioavailability of organic contaminants to degrading bacteria. Recent studies revealed that fungal hyphae have the potential to promote bacterial dispersal, and thus raised the idea of specifically stimulating the establishment of fungal networks in soils to increase contaminant bioavailability. Can such bacterial dispersal networks improve biodegradation performance considerably? If so, how are the improvements affected by abiotic conditions and by the spatial structure of dispersal networks? This doctoral thesis aims at answering these research questions. To this end, laboratory experiments are performed and a bacterial simulation model is developed, incorporating both microbiological and ecological theory. Manifold simulations and analyses of the microbial ecosystems’ spatiotemporal dynamics under different environmental scenarios reveal key factors and processes controlling biodegradation performance and determining benefits from bacterial dispersal networks.

Polychlorinated biphenyls (PCBs) are toxic and persistent chemicals that have been largely dispersed into the environment. The biological and abiotic transformations of PCBs often generate hydroxylated derivatives, which have been detected in a variety of environmental samples, including animal tissues and feces, water, and sediments. Because of their toxicity and widespread dispersion in the environment, hydroxylated PCBs (OH-PCBs) are today increasingly considered as a new class of environmental contaminants. Although PCBs are known to be susceptible to microbial degradation under both aerobic and anaerobic conditions, bacterial degradation of OH-PCBs has received little attention. The overall objective of this study is therefore to evaluate the transformation of mono-hydroxylated PCBs by the well characterized aerobic PCB-degrading bacterium, Burkholderia xenovorans LB400. In order to achieve our overall objective, a series of model mono-hydroxylated PCBs have been selected and they are used to determine the toxicity of hydroxylated congeners toward the bacterium B. xenovorans LB400. The biodegradation kinetics and metabolic pathways of the selected OH-PCBs by B. xenovorans LB400 are then characterized using GC/MS. To understand further the molecular basis of the metabolism of OH-PCBs by B. xenovorans LB400, gene expression analyses are conducted using reverse-transcription real-time (quantitative) polymerase chain reaction (RT-qPCR) and microarray technology. More formally, the specific aims of the proposed research are stated as follows: (1) To evaluate the toxicity of selected mono-hydroxylated derivatives of lesser-chlorinated PCBs toward the bacterium B. xenovorans LB400. (2) To assess the degradation of the selected OH-PCBs by B. xenovorans LB400. (3) To gain further understanding of the molecular bases of the metabolism of the selected OH-PCBs by B. xenovorans LB400. Three hydroxylated derivatives of 4-chlorobiphenyl and 2,5-dichlorobiphenyl, including 2'-hydroxy-, 3'-hydroxy-, and 4'-hydroxy- congeners, were significantly transformed by Burkholderia xenovorans LB400 when the bacterium was growing on biphenyl (biphenyl pathway-inducing conditions). On the contrary, only 2'-OH-4-chlorobiphenyl and 2'-OH-2,5-dichlorobiphenyl were transformed by the bacterium growing on succinate (conditions non-inductive of the biphenyl pathway). Gene expression analyses showed that only exposure to 2'-OH-4-chlorobiphenyl and 2'-OH-2,5-dichlorobiphneyl resulted in induction of key genes of the biphenyl pathway, when cells grown on succinate. These observations suggest that 2'OH-PCBs were capable of inducing the genes of biphenyl pathway. These results provide the first evidence that bacteria are able to cometabolize PCB derivatives hydroxylated on the non-chlorinated ring. Genome-wide transcriptional analyses using microarrays showed that 134 genes were differentially expressed in cells exposed to biphenyl, 2,5-dichlorobiphenyl, and 2'-OH-2,5-dichlorobiphneyl as compared to non-exposed cells. A significant proportion of differentially expressed genes were simultaneously expressed or down regulated by exposure to the three target compounds i.e., biphenyl, 2,5-DCB, and 2'-OH-2,5-DCB, which suggests that these structurally similar compounds induce similar transcriptional response of B.xenovorans LB400. Results of this study may have important implications for the natural attenuation of PCBs and fate of OH-PCBs in the environment. The recalcitrance to biodegradation and the high toxicity of some OH-PCBs may provide a partial explanation for the persistence of PCBs in the environment. / Civil Engineering

<p>Persistent organic pollutants (POPs) is a group of chemicals that are toxic, undergo long-range transport and accumulate in biota. Due to their persistency the distribution and recirculation in the environment often continues for a long period of time. Thereby they appear virtually everywhere within the biosphere, and poses a toxic stress to living organisms. In this thesis, attempts are made to contribute to the understanding of factors that influence the distribution of POPs with focus on processes in the marine environment. The bioavailability and the spatial distribution are central topics for the environmental risk management of POPs. In order to study these topics, various field studies were undertaken. To determine the bioavailable fraction of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated naphthalenes (PCNs), and polychlorinated biphenyls (PCBs) the aqueous dissolved phase were sampled and analysed. In the same samples, we also measured how much of these POPs were associated with suspended particles. Different models, which predicted the phase distribution of these POPs, were then evaluated. It was found that important water characteristics, which influenced the solid-water phase distribution of POPs, were particulate organic matter (POM), particulate soot (PSC), and dissolved organic matter (DOM). The bioavailable dissolved POP-phase in the water was lower when these sorbing phases were present. Furthermore, sediments were sampled and the spatial distribution of the POPs was examined. The results showed that the concentration of PCDD/Fs, and PCNs were better described using PSC- than using POM-content of the sediment. In parallel with these field studies, we synthesized knowledge of the processes affecting the distribution of POPs in a multimedia mass balance model. This model predicted concentrations of PCDD/Fs throughout our study area, the Grenlandsfjords in Norway, within factors of ten. This makes the model capable to validate the effect of suitable remedial actions in order to decrease the exposure of these POPs to biota in the Grenlandsfjords which was the aim of the project. Also, to evaluate the influence of eutrophication on the marine occurrence PCB data from the US Musselwatch and Benthic Surveillance Programs are examined in this thesis. The dry weight based concentrations of PCB in bivalves were found to correlate positively to the organic matter content of nearby sediments, and organic matter based concentrations of PCB in sediments were negatively correlated to the organic matter content of the sediment.</p>

A new complex aspect in the matter of water quality is the occurrence of emerging organic pollutants and contaminants in waste water. The currently low extent to which treatment of waste water is performed in Brazil, and in the world as a whole, there is a considerable need for development of cheap and accurate in-situ sampling methods for far-reaching studies of surface water quality. The lack of such methods today makes the maintenance and establishing of sanitary safety difficult. This diploma work gives a brief introduction to the basic principles of the passive sampling method known as Diffusive Gradient in Thin-films (DGT). A method that could be useful for such monitoring of quality in water bodies world wide. The aim of this study is to develop a method, for the detection of organic emerging pollutants and contaminants – i.e. compounds, which usually are present at very low concentrations when found in the environment as a result of human activity. More specifically, this work investigates the potential and usefulness of the application of DGT devices in detection of organic compounds that can affect human health and ecosystems, even at low concentrations, however, their effects still are in need of further investigations. This study focuses on both purely technical as well as practical points of views. The efficiency of organic DGT (o-DGT) with biochar as the adsorbing agent is examined targeting the detection of organic pollutants and contaminants in surface water. In this sense, the specific aim of the work is to evaluate the performance of biochar as adsorbing agent. This work showed that the performance of biochar as the adsorbing agent in binding layers in o-DGT sample devices can be considered as satisfactory since all compounds of interest in this study was successfully detected, quantified an identified. Further investigations in the future are needed to determine the effects of varying pH, temperature and ion concentration in the deployment media, as well as the properties of the binding layer in relation to concentration of biochar and the thickness of the layer. These in order to optimize the method for in-situ water sampling, aiming conventional use of biochar as the adsorbing agent in the future.

A poluição atmosférica já é considerada um caso de saúde pública nos grandes centros urbanos. Freqüentemente as emissões veiculares são as principais causas dessa poluição, principalmente as provenientes dos motores diesel, os quais, além da produção de material particulado (MP), em cuja superfície são adsorvidas substâncias carcinogênicas e mutagênicas, produzem poluentes como os hidrocarbonetos policíclicos aromáticos (HPAs). Contudo não existe uma avaliação direta dos riscos dessas emissões nos seres vivos. Plantas bioindicadoras podem dar idéia desses riscos. Este trabalho teve como objetivo avaliar o potencial de risco mutagênico da exaustão proveniente de um motor diesel, utilizando o bioensaio Trad-SH (clone KU-20) como bioindicador da poluição do ar. Nos experimentos, a exaustão do motor diesel aspirado de 2.0 L de deslocamento volumétrico foi diluído com ar atmosférico de modo a atingir concentrações de uma atmosfera pesadamente poluída (aproximadamente 50, 100 e 150 ppm de CO). Obtidos estes níveis de diluição, as inflorescências foram expostas a esta mistura de poluentes por duas horas (doses agudas). A concentração de CO foi monitorada continuamente por meio de um analisador de gases Horiba Enda utilizando o princípio de absorção seletiva no infravermelho. Para se avaliar o efeito mutagênico foi feita uma comparação entre as inflorescências não expostas aos poluentes (grupo 1) e as inflorescências expostas (grupos 2, 3 e 4) com aproximadamente 50, 100 e 150 ppm de CO respectivamente. Os dados obtidos foram analisados estatisticamente observando-se que, a freqüência média das mutações no grupo 1 (controle), foi significativamente mais baixa do que aquela dos grupos 3 e 4, porém foi similar à do grupo 2. Por sua vez, não houve diferenças significativas nas freqüências de mutações entre os grupos 3 e 4. Os resultados indicam que a exaustão do motor a diesel teve um papel significativo no desenvolvimento de mutações, mas somente quando diluída para concentrações de CO acima de 100 ppm. / Air pollution caused by vehicle emission has been considered as a major public health concern in the urban centers. Emission from diesel engine powered vehicles, in particular, are highly toxic, since carcinogenic and mutagenic compounds can adsorb on the expelled particles leading to the formation of the so-called polycyclic aromatic hydrocarbons (PAH\'s). A detailed investigation on the risks of those compounds on the living beings using the methodology applied in the work has not been carried out so far. This work is aimed at evaluating the mutagenic potential of the emission from a diesel engine using the Tradescantia stamen hair assay (TSH), by monitoring the stamen hair mutation in the clone KU-20. Experimentally, the inflorescence of the KU-20 clones was kept for 2 h under a simulated urban heavily polluted atmosphere, obtained by mixing the emission from a diesel engine and atmospheric air. The CO concentration in the atmosphere was monitored using a Horiba-Enda gas analyzer. The mutagenic effects of the atmosphere were analyzed by comparing a group of non-exposed control inflorescence (group 1) to inflorescences kept under polluted atmospheres containing 50, 100 and 150 ppm of CO, assigned to as groups 2, 3 and 4. The experimental data were analyzed using statistical methods. The frequency of mutations observed in the inflorescences from group 2 was slightly higher than that observed in the group 1. In the groups 3 and 4, however, the frequency of mutations was significantly higher than that exhibited by the control group. The latter suggests that the emission from a diesel engine plays an important role in the development of plants mutation, specially for atmospheres containing more than 100 ppm of CO.

O acúmulo de poluentes orgânicos persistentes é um dos principais problemas de contaminação do meio ambiente em todo o mundo. As atividades industriais e os avanços tecnológicos na maioria dos setores de produção contribuem com a crescente liberação de compostos poluentes, altamente tóxicos, biocumulativos e resistentes à degradação física, química, fotolítica e biológica. Esses poluentes são o resultado do uso indevido de pesticidas de um modo geral, da subprodução não planejada de sintéticos tóxicos como dioxinas policloradas e de uma lista enorme de atividades que promovem a combustão incompleta da matéria orgânica e despejam no ambiente uma vasta quantidade de hidrocarbonetos aromáticos persistentes. A biorremediação é uma estratégia biotecnológica que vem lançar luz sobre as técnicas de revitalização de sítios contaminados. É baseada na utilização de microorganismos, ou suas enzimas, para reduzir ou eliminar perigos ambientais contaminantes em substâncias inertes, CO2 e água. As oxigenases são uma classe de enzimas amplamente estudadas por suas propriedades catalíticas únicas, sendo a clorocatecol 1,2- dioxigenase de Pseudomonas putida um interessante alvo biotecnológico para a biorremediação devido a sua ampla especificidade a substratos aromáticos, aromáticos halogenados e dialogenados altamente poluentes, biocumulativos e carcinogênicos. Nesse trabalho, o protocolo de expressão heteróloga em E. coli foi implementado em nosso laboratório e a purificação realizada em coluna de afinidade através do uso de resina de quitina. Os cristais de cor marrom avermelhado da enzima clorocatecol 1,2-dioxigenase Pseudomonas putida foram obtidos na presença de polietilenogligol e acetato de magnésio durante o período de 10 dias, utilizando-se a técnica de difusão de vapor em gota sentada. A estrutura cristalográfica da enzima Pp 1,2-CCD foi determinada através da utilização da técnica MR-SAD, utilizando átomos de ferro, cofator da proteína, como agentes espalhadores e as coordenadas da enzima 3-clorocatecol 1,2-dioxigenase de Rhodococcus opacus como modelo de busca. O modelo final, contendo 3 moléculas na unidade assimétrica foi refinado a 3.4 Å de resolução. A enzima se enovela na forma dimérica (Fe3+)2, e apresenta de um modo geral, dois domínios catalíticos compostos de fitas-beta e uma região de loops e um domínio de ligação composto por hélices formando um túnel hidrofóbico. Como a primeira clorocatecol de bactérias gram-negativa a ser descrita para essa classe de enzimas, será possível investigar as diferenças conformacionais que levam aos mecanismos de catálise, amplo a diversos substratos, e avaliar características de seu funcionamento e ativação, como uma importante ferramenta para validação do papel desta proteína dentro do processo de biorremediação. / Accumulation of persistent organic pollutants is one of the most important environmental problems worldwide. Industrial activities and technological advances contribute to the spread of pollutants, highly toxic, bioaccumulative, and resistant to physical, chemical, photolytic and biological degradation. The persistent organic pollutants are consequence of the inappropriate use of pesticides, the production of toxic synthetic compounds such as polychlorinated biphenyls and a large list of activities promote incomplete combustion of which organic matter and release a large amount of persistent aromatic hydrocarbons to the environment. Bioremediation is a biotechnogical strategy that has shed light on revitalization techniques of contaminated sites. It is based on the application of microorganisms, or their enzymes, to eliminate or reduce environmental contaminants into inert substances, CO2 and water. The oxygenases are a class of largely studied enzymes due to their catalytic properties, being chlorocatechol 1,2-dioxygenase from Pseudomonas putida an interesting biotechnological target for bioremediation due to its specificity to a broad spectrum of highly polluted aromatic substrates. In the present work, heterologous expression protocol has been implemented in our laboratory and purification was performed by using affinity column in chitin resin. Brown-reddish crystals of chlorocatechol 1,2-dioxygenase from Pseudomonas putida were obtained in presence of polyethylene glycol (PEG) and magnesium acetate after 10 days, by utilizing vapor diffusion techniques in sitting drops. The crystallographic structure of Pp 1,2-CCD has been solved by MR-SAD technique, using iron atoms, the enzyme cofactor, as scattering centers and the coordinates of 3- chlorocatechol 1,2-dioxygenase of Rhodococcus opacus as the search model. The final model, containing three molecules in the asymmetric unit, has been refined up to 3.4 Å resolution. The enzyme folds in the dimeric form (Fe3+)2, and shows two catalytic domains composed of -strands and a loop region, and a helical domain that pack together forming a hydrophobic channel. As being the first member of the chlorocatechol dioxygenase family of enzymes from a gram-negative bacteria to be solved, it will be possible to explore the crystallographic structure of chlorocatechol 1,2- dioxygenase from Pseudomonas putida in order to investigate the conformational differences that explain its mechanism of action, on a broad spectrum of substrates, and evaluate the functional features, as an important tool to validate the role of this enzyme in the bioremediation process.