Download or read book Biodegradation and Molecular Analysis of 1 4 dioxane and Other Organic Contaminants in Soils Using Metagenomic Tools written by Vidhya Ramalingam and published by . This book was released on 2021 with total page 176 pages. Available in PDF, EPUB and Kindle. Book excerpt: Historically, 1,4-dioxane, a potential human carcinogen, was used as a stabilizer in 1,1,1-trichloroethane (1,1,1-TCA) formulations and is now frequently detected at chlorinated solvent contaminated sites. Bioremediation has emerged as an effective strategy to treat 1,4-dioxane. However, the distribution of 1,4-dioxane degrading species across various environmental samples is generally unknown. Additionally, 1,4-dioxane contamination typically occurs in groundwater under highly reducing conditions. There is a significant knowledge gap and a lack of information on the susceptibility of 1,4-dioxane to biodegradation under such reducing conditions. The success of organic contaminant bioremediation is often linked to the abundance of functional genes present in the soil that are associated with the degradation process. Although some information exists on the presence of these genes in contaminated soils, there is limited knowledge on the presence and diversity of these genes in uncontaminated soils. To address all these knowledge gaps, a series of studies were conducted. The first study aims at identifying which 1,4-dioxane degrading functional genes are present in soil communities and which genera may be using 1,4-dioxane and/or metabolites to support growth across different microbial communities. For this, laboratory sample microcosms and abiotic control microcosms (containing media) were inoculated with four uncontaminated soils and sediments from two contaminated sites. The sample microcosms were amended with 1,4-dioxane thrice and live control microcosms were treated in the same manner, except 1,4-dioxane was not added. Biodegradation was observed and whole genome shotgun sequencing was carried out. Although some degraders previously linked to 1,4-dioxane degradation were detected, Nocardioides, Gordonia and Kribbella were found to be potentially novel degraders. The functional genes associated with 1,4-dioxane demonstrated three genes were present at higher relative abundance values, including Rhodococcus sp. RR1 prmA, Rhodococcus jostii RHA1 prmA and Burkholderia cepacia G4 tomA3. The second study is focused on anaerobic biodegradation of 1,4-dioxane. The potential for 1,4-dioxane biodegradation was examined using multiple inocula and electron acceptor amendments. Compound specific isotope analysis (CSIA) was used to further investigate biodegradation in a subset of the microcosms. DNA was extracted from microcosms exhibiting 1,4-dioxane biodegradation for microbial community analysis using 16S rRNA gene amplicon high throughput sequencing. 1,4-dioxane biodegradation was most commonly observed in the nitrate amended and no electron acceptor treatments. However, it is important to note that the degradation was slow (approximately one year). The third study examines a set of genes associated with organic contaminant degradation in four uncontaminated (agricultural) soils. The abundance and diversity of benA, bph, dbfA, dxnA, etnC, etnE, ppaH, npaH, vcrA, xenA, xenB and xplA were investigated using protein sequences from the Functional Gene Pipeline and Repository (FunGene). The phylogenetic trees created indicate many genera may potentially be associated with each gene including Pseudomonas, Rhodococcus, Mycobacterium and Nocardioides. From these, some strains are well studied and are known to be involved in the biodegradation of organic contaminants and others are potentially new genera that may be associated with the biodegradation of the targeted group of contaminants.

Download or read book Stable Isotopic and Molecular Biological Tools to Validate Biodegradation of 1 4 dioxane written by Peerapong Pornwongthong and published by . This book was released on 2014 with total page 186 pages. Available in PDF, EPUB and Kindle. Book excerpt: 1,4-Dioxane, a probable human carcinogen, is a heterocyclic ether increasingly found as a contaminant in water supplies. Recent studies have reported that 1,4-dioxane can be biodegraded by a variety of microorganisms, and bioremediation may be an effective strategy for 1,4-dioxane contaminated sites. However, reliable monitoring tools to validate biodegradation of 1,4-dioxane are still lacking. Molecular biological tools and stable isotope-based tools have been previously applied as diagnostic tools for monitored natural attenuation and engineered bioremediation of various organic and inorganic compounds. In this study, molecular biological tools were used for determining bacterial populations, and for associating 1,4-dioxane biodegradation with relative copy numbers of phylogenetic and functional genes. These biomarkers were amplified using primers designed from the genome sequence data of 1,4-dioxane-degrading bacterium Pseudonocardia dioxanivorans CB1190, and were correlated with measured biodegradation rates. The results revealed that abundance of DXMO and 16S rRNA were in agreement with 1,4-dioxane biodegradation rates, and could be used to illustrate the inhibitory effect of co-contaminant transition metals Cu(II), Cd(II), Ni(II), and organic ligands such as tannic acid and L-cysteine. It should be recognized that biomarkers provide an indirect association between genes and enzyme activity. Factors regulating protein synthesis and catalytic activities of enzymes are not captured by nucleic acid-based biomarkers. This complicates the interpretation of biomarkers for predicting biodegradation rates. Compound specific isotope analysis (CSIA) could be used as another diagnostic tool to assess 1,4-dioxane biodegradation. In this study, hydrogen and carbon isotope analyses of 1,4-dioxane were successfully developed to determine isotope signatures of commercial 1,4-dioxane, and applied to determine kinetic isotope fractionation associated with biodegradation in both pure and mixed cultures, as well as abiotic degradation of 1,4-dioxane. During biodegradation, both 2H and 13C were enriched, while abiotic processes could enrich only 2H in residual 1,4-dioxane. This indicated that combined carbon and hydrogen isotope analyses of 1,4-dioxane allow differentiation of biological processes from abiotic mechanisms. Availability of stable isotopic and molecular biological tools will allow environmental engineering professionals to include bioremediation as an effective strategy in the cleanup of specific environmental contaminants.

Download or read book Microbial Ecology and Bioremediation of 1 4 Dioxane and Chlorinated Solvents Contaminated Groundwater and Soil written by Yu Miao and published by . This book was released on 2019 with total page 350 pages. Available in PDF, EPUB and Kindle. Book excerpt: It is crucial to consider the impact of abiotic and biological remediation technologies on the microbial ecology to predict the success of short-term active treatments and long-term passive attenuation processes. In this research, three bioremediation strategies were tested individually or coupled with chemical remedies in bench- and pilot-scale studies for removing 1,4-dioxane and chlorinated volatile organic compounds (CVOCs), which are widespread co-occurring contaminants in soils and water resources across the U.S., attracting attention because of their potential carcinogenicities. In each project, amplification of taxonomic and functional genes by qPCR as well as metagenomics including high-throughput sequencing were applied to provide reliable information about microbial communities in the ecological matrices as they transitioned from 1,4-dioxane and CVOC contaminations to exposures from treatment technologies and degradation products. A comprehensive multiple lines of evidence approach provided evidence of natural attenuation by microorganisms capable of metabolic or co-metabolic degradation of 1,4-dioxane within a large, diffuse plume. A pilot study of bioaugmentation with Pseudonocardia dioxanivorans CB1190 through direct injection as well as in-situ bioreactor was successfully conducted at a site impacted by 1,4-dioxane and CVOCs. Bench-scale microcosms were established to inform pilot-scale ex-situ bioreactors and in-situ propane biosparging at an industrial site. 1,4-Dioxane co-metabolism by indigenous microbes was accelerated by biostimulation with propane and nutrients. Inoculations with CB1190 or propanotroph, Rhodococcus ruber ENV425, were eventually outcompeted by native microbes, but gene allocations for xenobiotics and lipid metabolism were enhanced and accompanied rapid 1,4-dioxane degradation rates. Three synergistic treatment trains: oxidation & catalysis, oxidation & biodegradation, and catalysis & biodegradation, were applied to achieve nearly complete 1,4-dioxane removals even in the presence of inhibitory CVOCs. While oxidant- or nanocatalyst-tolerant microbes were dominant immediately after chemical processes, the microbial community thrived during the biodegradation in a deterministic process over time, presenting higher biodiversity that indicated a more stable community. The post-treatment community carried various functional potentials, such as degradation of CVOCs and aromatic hydrocarbons, as well as nitrogen fixation. These mechanistic and quantitative data will be valuable for developing synergistic treatments that lead to savings in cost, energy, and substrate amendments for the remediation of contaminant mixtures.

Download or read book Taxonomic and Functional Characterization of Microbial Communities Linked to Chlorinated Solvent 1 4 dioxane and Rdx Biodegradation in Groundwater and Soil Microcosms written by Hongyu Dang and published by . This book was released on 2021 with total page 221 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioremediation is becoming increasing popular for the remediation of sites contaminated with a range of different contaminants. Molecular methods such as 16S rRNA gene amplicon sequencing, shotgun sequencing, and high throughput quantitative PCR offer much potential for examining the microorganisms and functional genes associated with contaminant biodegradation, which can provide critical additional lines of evidence for effective site remediation. In this work, the first project examined the taxonomic and functional biomarkers associated with chlorinated solvent and 1,4-dioxane biodegradation in groundwater from five contaminated sites. Each site had previously been bioaugmented with the commercially available dechlorinating mixed culture, SDC-9. The results highlighted the occurrence of numerous genera previously linked to chlorinated solvent biodegradation. The functional gene analysis indicated two reductive dehalogenase genes (vcrA and tceA) from Dehalococcoides mccartyi were abundant. Additionally, aerobic and anaerobic biomarkers for the biodegradation of various chlorinated compounds were observed across all sites. The approach used (shotgun sequencing) is advantageous over many other methods because an unlimited number of functional genes can be examined and a more complete picture of the functional abilities of microbial communities can be depicted. Another research project evaluated the functional genes and species associated with RDX biodegradation at a RDX contaminated Navy site where biostimulation had been adopted. For this, DNA samples extracted from groundwater samples pre- and post-biostimulation were subject to shotgun sequencing and high throughput qPCR. DNA sequences from thirty-one RDX biodegraders were detected, with the most abundant species being Variovorax sp. JS1663. Further, nine RDX biodegrading species significantly (p

Download or read book Biodegradation of Dioxins and Furans written by Rolf-Michael Wittich and published by Springer Science & Business Media. This book was released on 1998-05-26 with total page 320 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioremediation of organic pollutants and heavy metals by use of microorganisms represents a safe, inexpensive, and environmentally-friendly concept in modern environmental engineering. During the last three decades intense efforts have been made by microbiologists and environmental engineers in the isolation and characterization of microorganisms capable of degradation, transformation and detoxification of recalcitrant chemical compounds of environmental concern: (polyhalogenated) dibenzo-p-dioxins, dibenzofurans, and diphenyl ethers. Special emphasis is placed on the potential of molecular biology techniques to improve presently available biocatalysts.

Download or read book Microbial Degradation of Xenobiotics written by Shree Nath Singh and published by Springer Science & Business Media. This book was released on 2011-10-07 with total page 488 pages. Available in PDF, EPUB and Kindle. Book excerpt: Our interest in the microbial biodegradation of xenobiotics has increased many folds in recent years to find out sustainable ways for environmental cleanup. Bioremediation and biotransformation processes harness the naturally occurring ability of microbes to degrade, transform or accumulate a wide range of organic pollutants. Major methodological breakthroughs in recent years through detailed genomic, metagenomic, proteomic, bioinformatic and other high-throughput analyses of environmentally relevant microorganisms have provided us unprecedented insights into key biodegradative pathways and the ability of organisms to adapt to changing environmental conditions. The degradation of a wide spectrum of organic pollutants and wastes discharged into the environment by anthropogenic activities is an emerging need today to promote sustainable development of our society with low environmental impact. Microbial processes play a major role in the removal of recalcitrant compounds taking advantage of the astonishing catabolic versatility of microorganisms to degrade or transform such compounds. New breakthroughs in sequencing, genomics, proteomics, bioinformatics and imaging are generating vital information which opens a new era providing new insights of metabolic and regulatory networks, as well as clues to the evolution of degradation pathways and to the molecular adaptation strategies to changing environmental conditions. Functional genomic and metagenomic approaches are increasing our understanding of the relative importance of different pathways and regulatory networks to carbon flux in particular environments and for particular compounds. New approaches will certainly accelerate the development of bioremediation technologies and biotransformation processes in coming years for natural attenuation of contaminated environments

Download or read book Biodegradation and Bioremediation written by Ajay Singh and published by Springer Science & Business Media. This book was released on 2013-03-09 with total page 332 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this volume, experts from universities, government labs and industry share their findings on the microbiological, biochemical and molecular aspects of biodegradation and bioremediation. The text covers numerous topics, including: bioavailability, biodegradation of various pollutants, microbial community dynamics, properties and engineering of important biocatalysts, and methods for monitoring bioremediation processes. Microbial processes are environmentally compatible and can be integrated with non-biological processes to detoxify, degrade and immobilize environmental contaminants.

Download or read book Environmental Investigation and Remediation written by Thomas K.G. Mohr and published by CRC Press. This book was released on 2010-03-25 with total page 550 pages. Available in PDF, EPUB and Kindle. Book excerpt: A ubiquitous, largely overlooked groundwater contaminant, 1,4-dioxane escaped notice by almost everyone until the late 1990s. While some dismissed 1,4-dioxane because it was not regulated, others were concerned and required testing and remediation at sites they oversaw. Drawing years of 1,4-dioxane research into a convenient resource, Environmental Investigation and Remediation: 1,4-Dioxane and other Solvent Stabilizers profiles the nature of 1,4-dioxane and several dozen other solvent stabilizer compounds. The author takes an approach he calls "contaminant archeology", i.e., reviewing the history of the contaminating chemical’s use in the industrial workplace at the site of release and how those uses impart chemical characteristics to the waste that affects its fate and transport properties. The book examines the uses, environmental fate, laboratory analysis, toxicology, risk assessment, and treatment of 1,4-dioxane in extensive detail. It provides case studies that document the contaminant migration, regulation, treatment, and legal aspects of 1,4-dioxane releases. It also describes the controversy over interpretation of 1,4-dioxane’s toxicology and associated risk, as well as the corresponding disparity in states’ regulation of 1,4-dioxane. A final chapter examines the policy implications of emerging contaminants like 1,4-dioxane, with discussion of opportunities to improve the regulatory and remedial response to this persistent contaminant in the face of toxicological uncertainty. Mobility, persistence, and treatment challenges combine to make 1,4-dioxane a particularly vexing contaminant. It is more mobile than any other contaminant you are likely to find at solvent release sites. Filled with case studies, equations, tables, figures, and citations, the book supplies a wide range of information on 1,4-dioxane. It then provides passive and active remediation strategies and treatment technologies for 1,4-dioxane in groundwater and provides you with the technical resources to help you decide which are appropriate for your site. For more information about Thomase Mohr and his book, go to http://www.The14DioxaneBook.com

Download or read book Biodegradation of 1 4 Dioxane in Co Contaminant Mixtures written by Shu Zhang and published by . This book was released on 2017 with total page 108 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioremediation is a promising technology to degrade or detoxify various organic and inorganic compounds in polluted environments by using microbiological activity, but it is sensitive to biogeochemical conditions as well as co-occuring compounds at impacted sites. This study focused on biodegradation of 1,4-dioxane, which is a carcinogen and an emerging water contaminant. 1,4-Dioxane was utilized as a stabilizer of chlorinated solvents, such as 1,1,1-trichloroethane (TCA); and it has been found widespread in groundwater. Many US states are implementing lower regulatory advisory levels based on the toxicity profile of 1,4-dioxane and the potential public health risks. However, the unique chemical properties of 1,4-dioxane, such as high water solubility, low Henry's law constant, and importantly, the co-occurrence with chlorinated solvents and other contaminants, increase the challenges to efficiently cleanup 1,4-dioxane contaminations. The objectives of this research were to measure and model the effects of chlorinated solvents on 1,4-dioxane metabolic biodegradation by laboratory pure cultures, elucidate the mechanisms of the inhibition, and test the effects of mixtures of co-contaminants in samples collected from actual 1,4-dioxane contaminated sites. It was determined that individual solvents inhibited biodegradation of 1,4-dioxane in the following order: 1,1-dichloroethene (1,1-DCE) > cis-1,2-diochloroethene (cDCE) > trichloroethene (TCE) > 1,1,1-trichloroethane (TCA). The results confirmed that 1,1-DCE was the strongest inhibitor of 1,4-dioxane biodegradation, even in chlorinated ethene mixtures. The energy production was delayed, and the genes coding for catalytic enzymes, dioxane monooxygenase (dxmB) and alcohol dehydrogenase (aldH) were down regulated, in the presence of chlorinated solvents. These results will be useful to scientists in understanding the fundamentals of enzymatic processes that catalyze biological degradation of hazardous compounds, and to environmental engineers by providing quantitative data valuable for the development of in-situ bioremediation approaches for contaminant mixtures.

Download or read book Biodegradation Pollutants and Bioremediation Principles written by Ederio Dino Bidoia and published by CRC Press. This book was released on 2021-04-20 with total page 398 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book presents a broad compendium of biodegradation research and discussions on the most up-to-date bioremediation strategies. The most relevant microbiological, biochemical and genetic concepts are presented alongside the fundamentals of bioremediation. The topics include: a wide variety of contaminant impacts evaluation, key methodologies required to measure biodegradation and propose new bioremediation protocols, as well as the handling of microbial communities related to such processes. The selected collaborating authors are renowned for their microbiology expertise and will provide an in-depth reference for students and specialists. The contents provide a valuable source of information for researchers, professionals, and policy makers alike.

Download or read book Microbial Biodegradation of Xenobiotic Compounds written by Young-Cheol Chang and published by CRC Press. This book was released on 2019-01-30 with total page 242 pages. Available in PDF, EPUB and Kindle. Book excerpt: Microbial Biodegradation of Xenobiotic Compounds examines and collects the recent information on the bioremediation technologies around the world. This book focuses on methods to decrease pollutants created by anthropogenic activities, industrial activities, and agricultural activities. This book answers some of the questions about – how to reduce contaminants? And whether there is a possibility of converting these pollutants in to useful energy by advanced biotechnological methods? The book combines present obtainable data with the expert knowledge of researchers from all over the world covering different aspects of environmental biotechnology and microbiology. It covers basic concepts of bioremediation and various methods involved in the bioremediation process, and provides specific chapters on the role of different genes and enzymes involved in microbial bioremediation process. It also gives special attention to heavy metal bioremediation by microalgae and the mechanisms involved during the degradation process. Recent innovative technologies about converting toxic pollutants in to useful energy like bioplastics and electricity are also discussed by specialist authors. Various chapters address the bioremediation of pesticides in soil using microbial metabolites, and molecular aspects of biodegradation which cover topics including identification of novel genes through the metagenomic approach and bioremediation using fungal laccase enzymes.

Download or read book Consequences of Microbial Interactions with Hydrocarbons Oils and Lipids Biodegradation and Bioremediation written by Robert J. Steffan and published by . This book was released on 2019 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: In this book international experts discuss the state-of-the-art in the biological degradation of hydrocarbons to meet remedial or disposal goals. The work focuses on practical applications, often on globally important scales including the remediation of some of the world's largest crude oil spills. Other related chapters discuss important implications of microbial transformation of hydrocarbons, including treatment of high fat processing wastes, impacts of microbial biodegradation activity on industrial processes, and the implications of microbial oil degradation in relation to modern oil extraction processes like hydraulic fracturing of shales and extraction of oil sands.

Download or read book Biodegradation of 1 4 Dioxane written by Yi Xiong and published by . This book was released on 2020 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: 1,4-Dioxane, a likely human carcinogen, has recently been detected in surface and groundwaters throughout the United States. As a contaminant of emerging concern, 1,4-dioxane has not yet been regulated by federal drinking water standards. The future regulatory limit for 1,4-dioxane is likely to be strict with federal maximum contaminant level lower than 1 μg/L. Therefore, 1,4-dioxane has stimulated great research interests. Conventional treatment processes including air stripping and carbon adsorption are not effective in removing 1,4-dioxane. Many advanced oxidation processes have demonstrated effectiveness at the expense of high energy and chemical costs. Biodegradation of 1,4-dioxane is an attractive alternative. However, 1,4-dioxane is generally recalcitrant to biodegradation, meaning that the growth rate of 1,4-dioxane degrading bacteria and the biodegradation rate of 1,4-dioxane is often slow. To promote the biodegradation of 1,4-dioxane, two approaches were investigated. In the first approach, a water film non-thermal plasma reactor and an aerobic biological reactor was combined to treat water contaminated by 1,4-dioxane. The plasma reactor converted 1,4-dioxane to organic products including ethylene glycol, ethylene glycol monoformate, ethylene glycol diformate, 1,4-dioxan-2-one, 1,4-dioxan-2-ol, and formic, oxalic, glycolic, and methoxyacetic acids. The concentration and composition of the organic products exiting the plasma reactor varied with hydraulic retention time (HRT) and affected the following biological treatment time. At the optimum HRT for the plasma reactor (i.e., 1 second), ~25% of 1,4-dioxane was mineralized to CO2, and ~75% was converted to organic products, which were further degraded in the following biological reactor. In the second approach, biodegradation of 1,4-dioxane with the addition of primary substrates was studied. Seven aqueous amendments (i.e., tetrahydrofuran, butanone, acetone, 1-butanol, 2-butanol, phenol and acetate) and five gaseous amendments (i.e., C1-C4 alkanes and ethylene) were evaluated as the primary substrates for dioxane degradation by mixed microbial consortia. The aqueous amendments were tested in microcosm bottles and the gaseous amendments were tested in a continuous-flow membrane biofilm reactor with hollow fibers pressurized by the gaseous amendments. Ethane was found to be the most effective gaseous substrate and tetrahydrofuran (THF) was the only aqueous substrate that promoted dioxane degradation. To understand the mechanisms by which THF affects dioxane biodegradation, changes in microbial community structure in microcosm experiments with synthetic media and landfill leachate were examined over time. The data revealed that in experiments with synthetic media, the addition of THF significantly increased the abundance of Pseudonocardia, a genus with several representatives that can grow on both dioxane and THF, and Rhodococus ruber, a species that can use THF as the primary growth substrate while co-metabolizing dioxane. However, in similar experiments with landfill leachate, only Rhodococcus ruber was significantly enriched. When the THF concentration was higher than the dioxane concentration, THF competitively inhibited dioxane degradation since dioxane degradation was negligible while the dioxane-degrading bacteria and the corresponding THF/dioxane monooxygenase gene copies increased by a few orders of magnitude. The effects of THF on 1,4-dioxane degradation was further investigated using two biological activated carbon reactors. The first reactor (denoted by Reactor H) treated dioxane at 500 [mu]g/L, a typical concentration in groundwater close to the contamination source. The second reactor (denoted by Reactor L) treated dioxane at 25 [mu]g/L at a comparable surface loading rate. The addition of THF promoted the dioxane degradation with an optimum THF: dioxane ratio of 3:1 in both reactors. In the reactor H, the relative abundance of Rhodococcus ruber increased from from 0.1% to 43.6% when the THF: dioxane ratio increased from 0:1 to 3:1, and decreased to 5.2% when the ratio further increased to 7:1. The influent dioxane concentration significantly affected the microbial community since Rhodococcus ruber was not enriched in the reactor L, and Mycobacterium was likely the microbe mainly responsible for dioxane degradation. The influent dioxane concentration also affected the mechanisms of dioxane degradation because the dioxane-monooxygenase and gene copies increased by 3 folds in the reactor H, but not in the reactor L. This dissertation is based on and adapted from published journal papers and manuscripts that are ready to submit.

Download or read book Investigations of Biodegradability and Toxicity of Organic Compounds written by Jan Dojlido and published by . This book was released on 1979 with total page 120 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Microbial Degradation of Organic Compounds written by David T. Gibson and published by . This book was released on 1984 with total page 562 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Biodegradation and Treatability of Specific Pollutants written by Edwin F. Barth and published by . This book was released on 1979 with total page 72 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Manual for Soil Analysis Monitoring and Assessing Soil Bioremediation written by Rosa Margesin and published by Springer Science & Business Media. This book was released on 2005-08-02 with total page 394 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume presents detailed descriptions of methods for evaluating, monitoring and assessing bioremediation of soil contaminated with organic pollutants or heavy metals. Traditional soil investigation techniques, including chemical, physical and microbiological methods, are complemented by the most suitable modern methods, including bioreporter technology, immunological, ecotoxicological and molecular assays. Step-by-step procedures, lists of required equipment and reagents and notes on evaluation and quality control allow immediate application