The technique of in situ hybridization is now vital to molecular biologists and their understanding of the pathophysiology of cellular functions. This practical guide covers all aspects of in situ hybridization, describing: practical procedures and protocols; the scientific background; areas of application; and the limitations of the technology. This edition has been completely rewritten to take into acocunt the many advances in the seven years since its initial publication.

This volume, written by a range of international experts, covers a wide range of topics involving organic fluorine compounds. Each chapter is preceded by a summary and includes extensive illustrations and references. The chapters cover atmospheric chemistry, application of 19F NMR, partition, degradation and transformation, naturally occurring organic fluorine compounds, toxicology of perfluoroalkanes and phosphorofluoridates, and application of aromatic compounds to the elucidation of the mechanism of cytochrome P450.

Diazotrophic bacteria convert atmospheric nitrogen to plant-useable form and this input of nitrogen through biological fixation is of great agronomic importance. The contributions presented in this volume relate to free-living nitrogen fixers and the diazotrophs associated with plants. Symbiotic association of Frankia with non-legumes and cyanobacterial associations are also discussed. Research topics covered in this volume include the biochemistry and genetics of diazotrophs, recent developments in improvement of plant-microbe interactions and their molecular basis, the use of molecular probes in taxonomy and ecology of diazotrophs and reports on field applications, agronomic importance and improvement in methodologies for assessing their contribution to plants. This book provides valuable information not only for researchers working in the field of biological nitrogen fixation but also for biochemistry, molecular biologists, microbiologists and agronomists.

The aim of this first book is to introduce the readers of the series to why Cuatro Cienegas Basin (CCB) is so unique, starting with the reason why astrobiologists became interested in this oasis in the first place; namely, the high diversity and abundance of stromatolites and microbial mats in continental waters to be found in the desert oasis. As NASA has long since discovered, the basin may offer the best analog of early Earth. In essence, CCB is a time machine that can take us far back and forth in time. In the respective chapters, the contributing authors explain the extraordinary microbial diversity of Cuatro Cienegas Basin from various perspectives. In order to do so, they explain their journey as well as the different tools used to unravel the basin's mysteries, such as: Why are there so many species in a place without food? How has life there survived the enormity of tectonic shifts through the ages, maintaining its ancient marine heritage?

Over the past 4 billion years, microorganisms have contributed to shaping the earth and making it more habitable for higher forms of life. They are remarkable in their metabolic diversity and their ability to harvest energy from oxidation and reduction reactions. Research on these microbiological processes has led to the newly evolving fields of geomicrobiology and biogeochemistry, linking the geosphere and the biosphere. This volume of the Soil Biology series provides an overview of the biogeochemical processes and the microorganisms involved, with an emphasis on the industrial applications. Topics treated include aspects such as bioremediation of contaminated environments, biomining, biotechnological applications of extremophiles, subsurface petroleum microbiology, enhanced oil recovery using microbes and their products, metal extraction from soil, soil elemental cycling and plant nutrition.

This book is a compilation of the research which was presented during the NATO-Advanced Research Workshop (ARW) entitled "Advances in Bacterial Paracrystalline Surface Layers" held in London, Ontario, Canada during September 27 to 30, 1992. The organizing committee consisted of the two Workshop directors, S. F. Kaval and T. J. Beveridge, and H. Konig, U. B. Sleytr and T. J. Trust; their summary statements about the significance and success of the NATO-ARWare in Chapter 37 of this book. This was the third international workshop on bacterial S-layers and it demonstrated unequivocally how rapidly research is progressing. The Workshop was made possible by financial support from the North Atlantic Treaty Organization (NATO), the Medical Research Council of Canada (MRC), the Natural Seiences and Engineering Research Council of Canada (NSERC), and the Canadian Bacterial Diseases Network (CBDN) which is a Canadian National Centre of Excellence (NCE). We are very grateful for the support from all of these agencies since their financial aid made it possible to bring to London, Canada a truly international group of S-layer experts. We encouraged the attendance and participation of graduate fellows and research associates, and their presentations students, postdoctoral was an intense three constitute the "Poster" section of this book. The NATO-ARW day workshop held at a delightful secluded location (Spencer Hall) so that the delegates had both formal and informal occasions to interact and evolve new ideas."

The Chlamydomonas Sourcebook, Third Edition, Volume Three: Cell Motility and Behavior has been fully revised and updated to include the wealth of new resources for the Chlamydomonas community. The book presents the latest advances in the area from an international array of expert authors, reflecting significant advancements in our understanding of the role of basal bodies and flagella in human diseases. In addition, employing quantitative proteomics/mass spectroscopy as well as cryo EM tomography and single particle cryo EM has revolutionized our knowledge of the axoneme in terms of the location of proteins and their interactions. Current insights on mitosis and cytokinesis, flagellar assembly and motility, intraflagellar transport, and more will ensure use of this reference as a guide for understanding human diseases of the cilium.

Two of the recent books in the Methods in Molecular Biology series, Yeast Protocols and Pichia Protocols, have been narrowly focused on yeasts and, in the latter case, particular species of yeasts. Food Microbiology Pro- cols, of necessity, covers a very wide range of microorganisms. Our book treats four categories of microorganisms affecting foods: (1) Spoilage organisms; (2) pathogens; (3) microorganisms in fermented foods; and (4) microorganisms p- ducing metabolites that affect the flavor or nutritive value of foods. Detailed information is given on each of these categories. There are several chapters devoted to the microorganisms associated with fermented foods: these are of increasing importance in food microbiology, and include one bacteriophage that kills the lactic acid bacteria involved in the manufacture of different foods-cottage cheese, yogurt, sauerkraut, and many others. The other nine chapters give procedures for the maintenance of lactic acid bacteria, the isolation of plasmid and genomic DNA from species of Lac- bacillus, determination of the proteolytic activity of lactic acid bacteria, det- mination of bacteriocins, and other important topics.

There is a growing global awareness of the link between good diet and health. This fascinating book reviews various functional foods or nutraceuticals and the bio-active compounds they contain in order to identify the role of bioactive compounds such as nisin, micronutrients, and hydrocolloids in the diet in overall human health. It also provides up-to-date information on functional elements like antioxidants, dietary fibres, pre & probiotics, vitamins and mineral-enriched foods in the human diet. Consisting of fifteen chapters, the book offers a systematic review of the key factors in the preparation of functional foods from selected sources, and also describes the processing, preservation and packaging of a range of functional food products. This book is a valuable resource for students and researchers working in the field of food science, food technology, and nutrition, as well as for industry experts.

The future of agriculture greatly depends on our ability to enhance productivity without sacrificing long-term production potential. The application of microorganisms, such as the diverse bacterial species of plant growth promoting bacteria (PGPB), represents an ecologically and economically sustainable strategy. The use of these bio-resources for the enhancement of crop productivity is gaining importance worldwide. "Bacteria in Agrobiology: Disease Management" discusses various aspects of biological control and disease suppression using bacteria. Topics covered include: fluorescent pseudomonads; siderophore-producing PGPR; pseudomonas inoculants; bacillus-based biocontrol agents; bacterial control of root and tuber crop diseases; fungal pathogens of cereals; soil-borne fungal pathogens; peronosporomycete phytopathogens; and plant parasitic nematodes.

This Volume addresses the pros and cons of oligonucleotide probes, primers and primer combinations, and importantly considers how to design the best tools for the microbial taxa and/or processes being investigated. Individual chapters focus on the design of primers targeting genes that code for enzymes associated with the following functions: degradation of aromatic, aliphatic and chlorinated hydrocarbons under aerobic and anaerobic conditions, methanogenesis, methane oxidation, and the nitrogen cycle. 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 book is an overview considering yeast and fermentation. The similarities and differences between yeasts employed in brewing and distilling are reviewed. The implications of the differences during the production of beer and distilled products (potable and industrial) are discussed. This Handbook includes a review of relevant historical developments and achievements in this field, the basic yeast taxonomy and biology, as well as fundamental and practical aspects of yeast cropping (flocculation), handling, storage and propagation. Yeast stress, vitality and viability are also addressed together with flavor production, genetic manipulation, bioethanol formation and ethanol production by non-Saccharomyces yeasts and a Gram-negative bacterium. This information, and a detailed account of yeast research and its implications to both the brewing and distilling processes, is a useful resource to those engaged in fermentation, yeast and their many products and processes.

This Volume presents relevant single-cell and single-molecule approaches in the study of microbes producing and utilizing hydrocarbons and lipids. While generically applicable for all microorganisms, the approaches described are, wherever possible, adapted to the field of study of hydrocarbon and lipid microbiology. The methods include basic procedures for isolating single cells by means of microfluidics and flow cytometry, and their cultivation in arrays as pure clones; for isolating, amplifying and sequencing single-cell genomes and transcriptomes; and for analysing single-cell metabolomes by means of Raman spectroscopy. Single-molecule approaches include the use of protein:fluorescent dye fusions for protein localization and methods for the production of cell division protostructures and lipid monolayers. Methods for the functional analysis of single cells include detection of metabolically active (protein-synthesizing) cells in environmental samples by bioorthogonal non-canonical amino acid tagging, Raman spectroscopy combined with stable isotope labelling and fluorescent in situ hybridisation, and visualization of single cells participating in gene transfer activity. Lastly, protocols are presented for single-cell biotechnological applications, including biofuel production. 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 is designed to be a long term career reference. The chapters present modern procedures. This is a how-to-book with a difference. These chapters:
- reveal the background information about working with salt loving organisms,
- are loaded with information about how experiments are conducted under high salt,
- provide information about analyses that work under these conditions and those that may not,
- present a wide range of details from laboratory designs to equipment used and even to simple anecdotal hints that can only come from experience.
Microbiological training focuses largely on the growth, the handling and the study of the microbes associated with humans and animals. Yet the largest proportion of the Earth s microbiota lives in saline environments such as the Oceans, saline deserts and terminal hypersaline environments. This need for salt can be intimidating for those interested in entering the field or for those interested in understanding how such research is accomplished."

Heavy metals always pose serious ecological risks when released into the environment due to their elemental non-degradable nature, regardless of their chemical form. This calls for the development of efficient and low-cost effluent treatment and metal recuperation technologies for contaminated waste water, not only because regulatory limits need to be met but also because the waste itself can be a resource for certain precious metals.

Biosorption is a general property of living and dead biomass to rapidly bind and abiotically concentrate inorganic or organic compounds from even very diluted aqueous solutions. As a specific term, biosorption is a method that utilizes materials of biological origin - biosorbents formulated from non-living biomass - for the removal of target substances from aqueous solutions. Recent research on biosorption provides a solid understanding of the mechanism underlying microbial biosorption of heavy metals and related elements.

This book gathers review articles analyzing current views on the mechanism and (bio)chemistry of biosorption, the performance of bacterial, fungal and algal biomass, and the practical aspects of biosorbent preparation and engineering. It also reviews the physico-chemical evaluations of biosorbents and modelling of the process as well as the importance of biosorption during heavy metal removal using living cells. It is a reference work for scientists, environmental safety engineers and R&D specialists who wish to further promote biosorption research and use the accumulated knowledge to develop and build industrial applications of biosorption in heavy metal separation technologies.

"

The aims of this book remain the same, that is, that it should be of in terest to all those people concerned with, or about, food hygiene in the broadest sense. There was clearly a need for a book of this sort and its success has necessitated a second edition. It will, I hope, answer criticisms that were justifiably made about certain omissions and shortcomings levelled at the earlier edition. The whole book has been thoroughly revised with the introduction of several new sections to various chapters. During the time that has elapsed since the earlier edition appeared there has been much publicity about newer forms of 'food poisoning'. Thus listeriosis is discussed in some detail whilst the problems of salmonellas in eggs and BSE are also considered. Interest in irradiated foods has waxed and waned but it is rightly included in the relevant chapter. There has been much progress in methodology with the advent of advanced molecular techniques such as gene probes and that of PCR; these are discussed briefly. I have included sections on HACCP which has come into great prominence in recent years thus answering a specific criticism made of the earlier edition. The chapter on water and waste disposal contains material on Legionnaires' disease and cryptosporidiosis, infections of much concern at the present time. Finally, the chapter on legislation has undergone a major revision with far greater emphasis being placed on EC food hygiene legislation."

Leishmania is a vector-borne pathogenic parasite found in 88 countries worldwide and is the causative agent of leishmaniasis. The different Leishmania species infect macrophages and dendritic cells of the host immune system, causing symptoms that range from disfiguring cutaneous and mucocutaneous lesions, widespread destruction of mucous membranes, or visceral disease affecting the haemopoetic organs. The recent publication of the complete genome sequences of three different Leishmania species provides new insights into this leading pathogen and presents scientists with an exciting resource to improve the understanding of its complex molecular and cellular biology. In this book, internationally recognized Leishmania experts critically review the most important aspects of current Leishmania research, providing the first coherent picture of the organism's molecular and cellular biology since the publication of the genome sequence. Chapters are written from a molecular and genomic perspective and discuss in depth Leishmania-specific aspects of trypanosomatid biology and pathology. Topics include: diagnosis and epidemiology, genome structure and content, regulation of gene expression, the Leishmania proteome, the Leishmania metabolome, Leishmania differentiation, interaction with the sand fly vector, drug discovery, drug resistance, and much more. This will be essential reading for all researchers working with Leishmania, trypanosomes, and protozoa; and is recommended for all biology and medical libraries.

Due to the highly collaborative nature of investigators working in the field, we have rapidly advanced our understanding of "Staphylococcus epidermidis "and other staphylococci in the last two decades. The chapters in "Staphylococcus Epidermidis: Methods and Protocols" are designed to give the new investigator a series of tools so they can ask novel and exciting questions related to the biology of this opportunistic pathogen, as many exciting and unexplored questions such as defining the interaction of "S. epidermidis "and other normal flora remain to be discovered. Written in the successful "Methods in Molecular Biology" series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, "Staphylococcus Epidermidis: Methods and Protocols" seeks to serve both professionals and novices with its well-honed methodologies.

A comprehensive guide to the revolutionary area of systems biology and its application in cell culture engineering, this volume presents an overall picture of the current topics central to structural and functional genomics, proteomics, metabolomics and bioinformatics, including such hot topics as RNAi, metabolic engineering and unfolded protein response. It includes reviews of the cellular response of environmental modulation such as low temperature and osmolarity, critical assessments of the applications of metablomics and fluxomics approaches, examination of the utility of modulation of key genes and a presentation of a theory of chemical organisation which provides new view on the system's structure. The clearly written chapters by experts in the field describe methods applicable to investigating the unique facets of cell culture.

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Fungal nanotechnology has great prospects for developing new products with industrial, agricultural, medicinal, and consumer applications in a wide range of sectors. The fields of chemical engineering, agri-food, biochemistry, pharmaceuticals, diagnostics, and medical device development all employ fungal products, with fungal nanomaterials currently used in applications ranging from drug development to the food industry and agricultural biotechnology. Fungal agents are an environmentally friendly, clean, non-toxic agent for the synthesis of metal nanoparticles and employ both intracellular and extracellular methods. The simplicity of scaling up and downstream processing and the presence of fungal mycelia which afford an increased surface area provide key advantages. In addition, the large spectrum of synthesized nanoparticle morphologies and the substantially faster biosynthesis rate in cell-free filtrate (due to the higher amount of proteins secreted in fungi) make this a particularly enticing route. Understanding the diversity of fungi in assorted ecosystems, as well as their interactions with other microorganisms, animals, and plants, underpins real and innovative technological developments and the applications of metal nanoparticles in many disciplines including agriculture, catalysis, and biomedical biosensors. Importantly, biogenic fungal nanoparticles show significant synergistic characteristics when combined with antibiotics and fungicides to offer substantially greater resistance to microbial growth and applications in nanomedicine ranging from topical ointments and bandages for wound healing to coated stents.

Coronaviruses were recognized as a group of enveloped, RNA viruses in 1968 and accepted by the International Committee on the Taxonomy of Viruses as a separate family, the Coronaviridae, in 1975. By 1978, it had become evident that the coronavirus genomic RNA was infectious (i. e., positive strand), and by 1983, at least the framework of the coronavirus replication strategy had been per ceived. Subsequently, with the application of recombinant DNA techniques, there have been remarkable advances in our understanding of the molecular biology of coronaviruses, and a mass of structural data concerning coronavirus genomes, mRNAs, and pro teins now exists. More recently, attention has been focused on the role of essential and accessory gene products in the coronavirus replication cyde and a molecular analysis of the structure-function relation ships of coronavirus proteins. Nevertheless, there are still large gaps in our knowledge, for instance, in areas such as the genesis of coronavirus subgenomic mRNAs or the function of the coronavirus RNA-dependent RNA polymerase. The diseases caused by coronaviruses have been known for much longer than the agents themselves. Possibly the first coronavirus-related disease to be recorded was feline infectious peritonitis, as early as 1912. The diseases associ ated with infectious bronchitis virus, transmissible gastroenteritis virus, and murine hepatitis virus were all well known before 1950."

Methane and its oxidation product, methanol, have occupied an important position in the chemical industry for many years: the former as a feedstock, the latter as a primary chemical from which many products are produced. More recently, the role played by methane as a potent "greenhouse" gas has aroused considerable attention from environmentalists and clima tologists alike. This role for C compounds has, of course, been quite 1 incidental to the myriad of microorganisms on this planet that have adapted their life-styles to take advantage of these readily available am bient sources. Methane, a renewable energy source that will always be with us, is actually a difficult molecule to activate; so any microorganism that can effect this may point the way to catalytic chemists looking for con trollable methane oxidation. Methanol, formed as a breakdown product of plant material, is also ubiquitous and has also encouraged the growth of prokaryotes and eukaryotes alike. In an attempt to give a balanced view of how microorganisms have been able to exploit these simple carbon sources, we have asked a number ofleading scientists (modesty forbids our own inclusion here) to contribute chapters on their specialist areas of the subject."