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Books > Professional & Technical > Biochemical engineering > General
This Volume describes methods for investigating microbes in their natural environment and how to obtain representative samples and preserve them for subsequent analyses. Chapters are arranged according to the environments under investigation, which include: oil reservoirs, fracking fluids, aquifers, coal beds, oil sands and their tailing ponds, lakes, rivers, leaves, polar seas and ice, the sea-surface microlayer, mud flats, microbialites, and deep-sea fauna. A variety of downstream analytical procedures are described, including: nucleic-acid extraction and preparation for high-throughput sequencing, fluorescence in-situ hybridisation, and cultivation of aerobic and anaerobic hydrocarbon-degrading microbes. Though most chapters focus on hydrocarbon-rich environments, many of the approaches used are generic, and as such will be of value to researchers embarking on studies of microbes and their processes in the field. Hydrocarbon and Lipid Microbiology Protocols There are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
The series Topics in Heterocyclic Chemistry presents critical reviews on present and future trends in the research of heterocyclic compounds. Overall the scope is to cover topics dealing with all areas within heterocyclic chemistry, both experimental and theoretical, of interest to the general heterocyclic chemistry community. The series consists of topic related volumes edited by renowned editors with contributions of experts in the field. All chapters from Topics in Heterocyclic Chemistry are published Online First with an individual DOI. In references, Topics in Heterocyclic Chemistry is abbreviated as Top Heterocycl Chem and cited as a journal.
This book addresses different aspects of green biocomposite manufacture from natural fibres and bioplastics, including the manufacturing procedures and the physical, mechanical, thermal and electrical properties of green biocomposites. Featuring illustrations and tables that maximize reader insights into the current research on biocomposites, it emphasises the role of green technology in the manufacture of biocomposites and analysis of properties of biocomposites for different applications. It is a valuable resource for researchers and scientists in industry wanting to understand the need for biocomposites in the development of green, biodegradable and sustainable products for different applications.
This book offers an up-to-date overview of the concepts, modeling, technical and technological details and practical applications of different types of sensors, and discusses the trends of next generation of sensors and systems for environmental and food engineering. This book is aimed at researchers, graduate students, academics and industry professionals working in the field of environmental and food engineering, environmental monitoring, precision agriculture and food quality control.
This book serves to highlight the seamless integration of the sciences leading to sustainable technologies. Chemical engineering is one of the major disciplines catering to the societal needs in the fields of energy, environment and materials. The chapters of this book have been selected to encompass the latest in industrial biotechnology and biochemical engineering principles and applications. The chapters are included here after careful review for content and depth. The book focuses on the relatively new areas of molecular biotechnology and nanotechnology which have a strong impact at the fundamental and process levels in chemical engineering. The book also covers analytical procedures, experimental techniques and process analysis in bioprocessing, bioremediation, green separation methods, and emerging nanoparticle applications. It should be useful to students, academicians, and practitioners alike.
This book provides a critical, carefully researched, up-to-date summary of membranes for membrane bioreactors. It presents a comprehensive and self-contained outline of the fundamentals of membrane bioreactors, especially their relevance as an advanced water treatment technology. This outline helps to bring the technology to the readers' attention, and positions the critical topic of membrane fouling as one of the key impediments to its more widescale adoption. The target readership includes researchers and industrial practitioners with an interest in membrane bioreactors.
The book introduces the outcomes of latest research in the field of Chemical Engineering. The book also illustrates the application of Chemical Engineering principles to provide innovative and state of the art solutions to problems associated with chemical industries. It covers a wide spectrum of topics in the area of Chemical Engineering such as Transfer operations, novel separation processes, adsorption, photooxidation, process control, modelling, and simulation. The book provides timely contribution towards implementation of recent approaches and methods in Chemical Engineering Research. It presents chapters focussed on several Chemical Engineering principles and methodologies of wide multidisciplinary applicability. The intended audience of this book will mainly consist of researchers, research students, and practitioners in Chemical Engineering and allied fields. The book can also serve researchers and students involved in multidisciplinary research.
This book covers the latest developments in enzyme immobilization with its wide applications, such as for industry, agriculture, medicine, and the environment. Topics covered include basics of enzyme immobilization, its implication in therapeutics and disease diagnostics, and its significance in solving environmental problems. This is an ideal book for researchers, graduate and postgraduate students, as well as scientists in industry, agriculture and health sectors. This book is a complete summary of enzyme immobilization and also thoroughly covers all the latest research. This book covers: The last one-hundred years of innovative research done in enzyme immobilization Recent developments in immobilization techniques, such as types of matrices, immobilization methods, and linking agents, as well as enzyme immobilization without any matrices and its properties The physiological and industrial significance of enzymes from plants and the implementation of immobilized enzymes in the treatment of waste water and polluted air Biomedical and bioanalytical applications of immobilized enzymes
This book delves into the recent developments in the microscale and microfluidic technologies that allow manipulation at the single and cell aggregate level. Expert authors review the dominant mechanisms that manipulate and sort biological structures, making this a state-of-the-art overview of conventional cell sorting techniques, the principles of microfluidics, and of microfluidic devices. All chapters highlight the benefits and drawbacks of each technique they discuss, which include magnetic, electrical, optical, acoustic, gravity/sedimentation, inertial, deformability, and aqueous two-phase systems as the dominant mechanisms utilized by microfluidic devices to handle biological samples. Each chapter explains the physics of the mechanism at work, and reviews common geometries and devices to help readers decide the type of style of device required for various applications. This book is appropriate for graduate-level biomedical engineering and analytical chemistry students, as well as engineers and scientists working in the biotechnology industry.
Sensors were developed to detect and quantify structures and functions of human body as well as to gather information from the environment in order to optimize the efficiency, cost-effectiveness and quality of healthcare services as well as to improve health and quality of life. This book offers an up-to-date overview of the concepts, modeling, technical and technological details and practical applications of different types of sensors. It also discusses the trends for the next generation of sensors and systems for healthcare settings. It is aimed at researchers and graduate students in the field of healthcare technologies, as well as academics and industry professionals involved in developing sensing systems for human body structures and functions, and for monitoring activities and health.
This Volume presents protocols for systems and synthetic biology applications in the field of hydrocarbon and lipid microbiology. It complements another Volume that describes generic protocols for wet experimental and computer-based systems and synthetic biology studies. The protocols in this Volume demonstrate how to employ systems and synthetic biology approaches in the design of microbes for the production of esters, isoprenoids, hydrophobic polymers, rhamnolipid biosurfactant, and peptide antimicrobial and thioether-stabilised molecules. Also presented is a protocol for the engineering of transcription factor-based biosensors for intracellular products, and another for the creation of a synthetic hydroxylase with novel activity for the selective oxyfunctionalisation of linear alkanes. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
This Volume provides protocols for the biochemical analysis of hydrocarbon- and lipid-relevant products, cell components and activities of microbes that interact with hydrophobic compounds. They include methods for the extraction, purification and characterisation of surface tension-reducing bioemulsifiers and biosurfactants that increase the surface area and hence bioavailability of hydrophobic substrates. Protocols for the isolation and biochemical analysis of lipids and polyhydroxyalkanoates, food storage products made during nutrient abundance that represent important biotechnological products, are presented. The extraction of membrane lipid rafts, sub-organelles that fulfil important functional roles for the cell membrane, and the isolation and characterisation of membrane phospholipid biomarkers, are also described. The purification and characterisation of integral membrane hydrocarbon-oxidising enzymes are addressed. Lastly, two generic methods for the genetic analysis of catabolic pathways and analysis of ligand binding are presented. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
This Volume covers protocols for in-silico approaches to hydrocarbon microbiology, including the selection and use of appropriate statistical tools for experimental design replication, data analysis, and computer-assisted approaches to data storage, management and utilisation. The application of algorithms to analyse the composition and function of microbial communities is presented, as are prediction tools for biodegradation and protein interactions. The basics of a major open-source programming language, Python, are explained. Protocols for calculating reaction kinetics and thermodynamics are presented, and modelling the environmental fate of hydrocarbons during bioremediation is explained. With the exception of molecular biology studies of molecular interactions, the use of statistics is absolutely essential for both experimental design and data analysis in microbiological research, and indeed in the biomedical sciences in general. Moreover, studies of highly varying systems call for the modelling and/or application of theoretical frameworks. Thus, while two protocols in this Volume are specific to hydrocarbon microbiology, the others are generic, and as such will be of use to researchers investigating a broad range of topics in microbiology and the biomedical sciences in general. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
This thesis explores the ability of M. maripaludis to capture and convert CO2 to methane in the presence of free nitrogen, and offers a consolidated review of the metabolic processes and applications of M. maripaludis. Further, it develops, validates and analyzes the first genome-scale metabolic model (iMM518) of M. maripaludis. Readers will discover, for the first time, the impact of nitrogen fixation on methane production. As such, the thesis will be of interest to researchers working on M. maripaludis, biofuels and bioenergy, systems biology modeling and its experimental validation, estimation of maintenance energy parameters, nitrogen fixing microbes, and bioremediation.
This Volume presents generic protocols for wet experimental and computer-based systems and synthetic biology approaches relevant to the field of hydrocarbon and lipid microbiology. It complements a second Volume that describes protocols for systems and synthetic biology applications. The wet experimental tools presented in this Volume include protocols for the standardisation of transcriptional measurements, application of uracil excision-based DNA editing for, inter alia, multi-gene assembly, the use of fluxomics to optimise "reducing power availability", and the incorporation of non-canonical amino acids into proteins for optimisation of activities. Phenome-ing microbes, using a combination of RNA-seq and bioinformatic algorithms, is presented, as is an illustration, using methylotrophs as an example, of how the different key omics approaches constitute a pipeline for functional analysis, acquisition of a systems overview, and metabolic optimisation. Complementary computational tools that are presented include protocols for probing the genome architecture of regulatory networks, genome-scale metabolic reconstruction, and bioinformatic approaches to guide metabolic engineering. The Volume also includes an overview of how synthetic biology approaches can be used to improve biocontainment. Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
A unique feature of this book is its focus on nanotechnological solutions for the production of bioenergy and biofuels. Coverage includes topics such as nanobiotechnology, microalgae, biofuel cells, biomass pretreatment, and biomass conversion. An international team of experts also addresses the need to precisely characterize nanoparticles and the role of catalysts. The range of topics addressed, together with a chapter on risk management, make this book a highly useful resource for a broad readership including physicists, chemists, microbiologists, biotechnologists, food technologists, agricultural engineers, and nanotechnologists.
This Volume presents methods for quantifying microbial populations and characterising microbial communities by extracting and analysing biomarkers such as RNA, DNA and lipids. The chapters cover a wide range of topics, including: cell separation from oil-rich environments, enumeration of hydrocarbon degraders and sulphate reducers using most-probable-number techniques, and quantification by means of real-time PCR. A variety of molecular methods are described for microbial community profiling, such as phospholipid fatty acid analysis, DGGE, T-RFLP and SSCP. One chapter examines high-throughput sequencing, and provides important information on the associated procedures required for thorough data analysis. A further chapter is devoted to the characterisation of protistan communities, while the closing chapter describes multiplex fluorescent antibody microarrays for detecting microbial biomarkers. Hydrocarbon and Lipid Microbiology Protocols There are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
This Volume features protocols for investigating the hydrocarbon- and lipid-specific activities of microbes. They include methods for studying chemotaxis, the colonisation of hydrocarbon surfaces, hydrocarbon uptake, respiration, nitrogen fixation, sulphate reduction, membrane stabilisation through cis-trans isomerisation of membrane fatty acids, and the production of biosurfactants and biopolymers in response to the presence of hydrocarbons. A protocol for studying the ability of microbes to control the concentration of hydrocarbons in their aqueous environment is also described, and phenotyping methods to reveal microbes' more general metabolic activities are presented. Several protocols for investigating acid production in connection with oil souring and biocorrosion by microbes in oil well, oil transportation and storage settings are presented. Lastly, protocols for measuring methanogenesis, as an example of microbial hydrocarbon production, are described.< Hydrocarbon and Lipid Microbiology ProtocolsThere are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
This book covers the latest developments in rolling circle amplification (RCA) technology with applications in clinical diagnostic tests and molecular medicine. Topics covered include new enzymes useful in RCA, techniques involving RCA for enhanced signal amplification, novel RCA diagnostics, sensors for expediting RCA detection, and prospective RCA-based therapeutics. This is a valuable book for university professors and students in the field of biomedical engineering and biomolecular pharmacology as well as R&D managers of biotechnology and biopharmaceutical companies. Specifically, this book: Reviews prospective RCA-based therapeutics, including RCA-derived DNA nanoparticles that strongly bind to cancer cells Expands readers' understanding of sensor systems for expediting detection of RCA products by using probe-tagged magnetic nanobeads Maximizes reader insights into novel RCA diagnostics, such as PNA openers-assisted RCA for detection of single target cells and in situ RCA diagnosis of cancer cells and malignant tissues Presents innovative methods for quasi-exponential enhancement of RCA-generated signals, such as nicking enzyme-assisted cascade RCA and RCA coupled with loop-mediated amplification Advance Praise for Rolling Circle Amplification (RCA): "This book provides a badly needed compendium of innovative RCA methods and applications. It should help further increase the community of scientists that have employed RCA in research and diagnostic programs."- Charles Cantor, Professor Emeritus of Biomedical Engineering, Boston University Executive Director, Retrotope Inc. (USA) "In this new book Vadim Demidov has assembled an enticing menu of articles that illustrate the evolution of the RCA field, including improved protein parts for building superior DNA nanomachines, enhanced modalities of amplification and detection, diagnostic applications, and even a sampling of potential therapeutic applications. The reader will appreciate that while RCA has come of age, there is no lack of exciting surprises, turns, and twists in the continuing evolution of the technology."- Paul Lizardi, Professor of Pathology, Yale University School of Medicine (retired) Investigator, University of Granada, Spain, President, PetaOmics, Inc., San Marcos, Texas.
This Volume presents applications of hydrocarbon microbiology in the context of environmental pollutant degradation, covering pollutants such as petroleum and related wastes (i.e. oil sludge), biofuels, lipid-rich wastes, chlorinated solvents and BTEX, in several environments (marine, soil, groundwater). The approaches presented range from laboratory experiments and treatment in reactors to field applications. Two chapters highlight innovative approaches to address relevant questions in pollutant degradation, such as low environmental concentrations of pollutants, and the biodegradation of complex pollutant mixtures using biofilms. Rather than presenting the applications in the form of protocols, some of the chapters in this Volume include detailed practical information on the opportunities offered by and limitations of the different approaches, providing valuable information for researchers planning to perform bioremediation experiments. Hydrocarbon and Lipid Microbiology Protocols There are tens of thousands of structurally different hydrocarbons, hydrocarbon derivatives and lipids, and a wide array of these molecules are required for cells to function. The global hydrocarbon cycle, which is largely driven by microorganisms, has a major impact on our environment and climate. Microbes are responsible for cleaning up the environmental pollution caused by the exploitation of hydrocarbon reservoirs and will also be pivotal in reducing our reliance on fossil fuels by providing biofuels, plastics and industrial chemicals. Gaining an understanding of the relevant functions of the wide range of microbes that produce, consume and modify hydrocarbons and related compounds will be key to responding to these challenges. This comprehensive collection of current and emerging protocols will facilitate acquisition of this understanding and exploitation of useful activities of such microbes.
This textbook teaches advanced undergraduate and first-year graduate students in Engineering and Applied Sciences to gather and analyze empirical observations (data) in order to aid in making design decisions. While science is about discovery, the primary paradigm of engineering and "applied science" is design. Scientists are in the discovery business and want, in general, to understand the natural world rather than to alter it. In contrast, engineers and applied scientists design products, processes, and solutions to problems. That said, statistics, as a discipline, is mostly oriented toward the discovery paradigm. Young engineers come out of their degree programs having taken courses such as "Statistics for Engineers and Scientists" without any clear idea as to how they can use statistical methods to help them design products or processes. Many seem to think that statistics is only useful for demonstrating that a device or process actually does what it was designed to do. Statistics courses emphasize creating predictive or classification models - predicting nature or classifying individuals, and statistics is often used to prove or disprove phenomena as opposed to aiding in the design of a product or process. In industry however, Chemical Engineers use designed experiments to optimize petroleum extraction; Manufacturing Engineers use experimental data to optimize machine operation; Industrial Engineers might use data to determine the optimal number of operators required in a manual assembly process. This text teaches engineering and applied science students to incorporate empirical investigation into such design processes. Much of the discussion in this book is about models, not whether the models truly represent reality but whether they adequately represent reality with respect to the problems at hand; many ideas focus on how to gather data in the most efficient way possible to construct adequate models. Includes chapters on subjects not often seen together in a single text (e.g., measurement systems, mixture experiments, logistic regression, Taguchi methods, simulation) Techniques and concepts introduced present a wide variety of design situations familiar to engineers and applied scientists and inspire incorporation of experimentation and empirical investigation into the design process. Software is integrally linked to statistical analyses with fully worked examples in each chapter; fully worked using several packages: SAS, R, JMP, Minitab, and MS Excel - also including discussion questions at the end of each chapter. The fundamental learning objective of this textbook is for the reader to understand how experimental data can be used to make design decisions and to be familiar with the most common types of experimental designs and analysis methods.
Artificial organs may be considered as small-scale process plants, in which heat, mass and momentum transfer operations and, possibly, chemical transformations are carried out. This book proposes a novel analysis of artificial organs based on the typical bottom-up approach used in process engineering. Starting from a description of the fundamental physico-chemical phenomena involved in the process, the whole system is rebuilt as an interconnected ensemble of elemental unit operations. Each artificial organ is presented with a short introduction provided by expert clinicians. Devices commonly used in clinical practice are reviewed and their performance is assessed and compared by using a mathematical model based approach. Whilst mathematical modelling is a fundamental tool for quantitative descriptions of clinical devices, models are kept simple to remain focused on the essential features of each process. Postgraduate students and researchers in the field of chemical and biomedical engineering will find that this book provides a novel and useful tool for the analysis of existing devices and, possibly, the design of new ones. This approach will also be useful for medical researchers who want to get a deeper insight into the basic working principles of artificial organs.
This book connects a retrosynthetic or disconnection approach with synthetic methods in the preparation of target molecules from simple, achiral ones to complex, chiral structures in the optically pure form. Retrosynthetic considerations and asymmetric syntheses are presented as closely related topics, often in the same chapter, underlining the importance of retrosynthetic consideration of target molecules neglecting stereochemistry and equipping readers to overcome the difficulties they may encounter in the planning and experimental implementation of asymmetric syntheses. This approach prepares students in advanced organic chemistry courses, and in particular young scientists working at academic and industrial laboratories, for independently solving synthetic problems and creating proposals for the synthesis of complex structures.
This book covers the state-of-the-art research on molecular biology assays and molecular techniques enabled or enhanced by microfluidic platforms. Topics covered include microfluidic methods for cellular separations and single cell studies, droplet-based approaches to study protein expression and forensics, and microfluidic in situ hybridization for RNA analysis. Key molecular biology studies using model organisms are reviewed in detail. This is an ideal book for students and researchers in the microfluidics and molecular biology fields as well as engineers working in the biotechnology industry. This book also: Reviews exhaustively the latest techniques for single-cell genetic, epigenetic, metabolomic, and proteomic analysis Illustrates microfluidic approaches for inverse metabolic engineering, as well as analysis of circulating exosomes Broadens readers' understanding of microfluidics convection-based PCR technology, microfluidic RNA-seq, and microfluidics for robust mobile diagnostics
This book presents the latest advances in and current research perspectives on the field of urban/industrial solid waste recycling for bio-energy and bio-fuel recovery. It chiefly focuses on five main thematic areas, namely bioreactor landfills coupled with energy and nutrient recovery; microbial insights into anaerobic digestion; greenhouse emission assessment; pyrolysis techniques for special waste treatment; and industrial waste stabilization options. In addition, it compiles the results of case studies and solid waste management perspectives from different countries. |
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