Bacterial Spores: Current Research and Applications | Book
"well-written and edited" (Microbiol. Today)
Caister Academic Press
Department of Chemistry, University of Nevada, Las Vegas, USA
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Bacterial endospores are dormant structures produced by stressed bacterial cells. Due to their intrinsic resistance endospores are perfect vehicles for causing infection. A number of endospore producing bacteria can cause diseases including tetanus, anthrax and botulism; other spore forming species have been used in biotechnological applications such as probiotics and biocides. Modern molecular studies are providing new insights into the biochemical and biophysical characteristics of spore forming bacteria that may lead to promising new applications, detection methods and therapeutics.
In this book expert authors from around the world contribute comprehensive, up-to-date reviews on the current state of our knowledge of bacterial endospores. Topics covered include: gene regulation of sporulation, fruiting body development in Myxococcus xanthus, sporulation in Streptomyces, structure and composition of the bacterial spore, mechanisms of spore survival, germination of spores, spore peptidoglycan degradation, water and cations flux during sporulation and germination, the spore as an infectious agent, heterogeneity in spore populations, detection of bacterial spores, and the expression of recombinant proteins using spores.
An essential text for everyone involved in spore research, the expression of recombinant proteins and pathogen detection, this book is also recommended for all scientists that like to keep up with cutting-edge research in microbiology and biotechnology.
"well-written and edited chapters on bacterial sporulation, with most chapters being relatively short, but comprehensive ... This is an imaginatively edited book with up-to-date references and a sensible index. It is suitable both for the specialist and general reader." from Microbiology Today (2013)
Table of contents
1. Historical Notes and Introduction to Bacterial Spores
Mark Torred, Elias Benjelloun, Eramelle Dibala, Ernesto Abel-Santos and Christian Ross
2. Gene Regulation of Sporulation in Bacillus subtilis
Eduardo A. Robleto, Holly A. Martin, Amber M. Pepper and Mario Pedraza-Reyes
Endospore formation in the Bacilli has been used extensively as a proxy for cell growth and differentiation. Research that spans many years has revealed that endospore formation is a complex and elegant process that exquisitely coordinates the development of a dormant spore. The spore structure allows survival when nutrients and moisture are scarce. Once growth-conducive conditions are restored, the process of germination gives rise to vegetative cells. The cell differentiation program of sporulation is activated upon cessation of growth. Comparison of gene regulons activated during sporulation and stress survival suggest that the mode of regulation of endospore formation is well conserved between Bacilli and Clostridia.
3. Fruiting Body Development in Myxococcus xanthus: a Multicellular Developmental Program That Leads to Sporulation
Krista M. Giglio and Anthony G. Garza
The myxobacteria are Gram-negative spore formers. Although the spores of myxobacteria and those of Bacillus species, which are Gram-positive bacteria, have many properties in common, the developmental processes that lead to their formation are fundamentally different. In this chapter, we give a general overview of sporulation in the model myxobacterium Myxoccocus xanthus. We discuss the developmental process that yields M. xanthus spores, the structural and biochemical properties of the spores and the genes that are associated with sporulation in this organism, and we compare each to its counterpart in Bacillus.
4. Streptomyces Sporulation
Julia P. Swiercz and Marie A. Elliot
Streptomyces are soil-dwelling Gram positive bacteria with a complex, multicellular life cycle. The latter stages of their life cycle are defined by the metamorphosis of multi-genomic aerial hyphae into chains of unigenomic exospores. Here, we discuss the classical studies that established a solid genetic understanding of aerial development and sporulation, and highlight important new advances in the areas of cell division and spore septum placement, chromosome segregation and condensation during sporulation, and spore maturation.
5. The Structure and Composition of the Outer Layers of the Bacterial Spore
Arthur I. Aronson
This review summarizes the basic structural features, composition and protein interactions of the bacterial spore. Aspects of the possible interactions between the external layers of the bacterial spore, the coat and exosporium and the functions of each is discussed in detail. An important theme is that spores are not simply dormant resistant structures for long term survival under conditions unsuitable for germination. Rather they are adaptable to physical and nutritional conditions in order to enhance their dormancy (resistance), infectivity and dispersal with the spore coat and exosporium having significant roles. Multiple functions beyond protection of the spore cytoplasm, a role in germination and attachment to substrata can begin to account for the complexity of each of these structures.
6. Mechanisms of Bacterial Spore Survival
Mario Pedraza-Reyes, Norma Ramírez-Ramírez, Luz E. Vidales-Rodríguez and Eduardo A. Robleto
Sporulation is the process by which a spore is formed and is generally induced when microorganism of the Bacilli and Clostridia genus are challenged by nutritional stress. Spores of Bacillus species contaminate food and other products of human consumption compromising health. Therefore, a deeper understanding of the mechanisms involved in long-term survival is important to develop rational strategies for spore killing. Spores can survive for extended periods of time and possess the ability to return to life if nutrients become available. For decades bacterial spores have served as extraordinary paradigms to understand the consequences of long-term exposure to environmental factors that potentially compromise cell viability. This special type of cells can withstand a wide range of assaults that would otherwise destroy its vegetative cell counterparts. Spores of the genus Bacillus are several times more resistant to heat, desiccation, UV-C light, oxidizing chemicals and other genotoxic drugs. The particular cell architecture of spores and multiple mechanisms to protect its genome from citotoxic and genotoxic factors explain in great part such resistance. The present chapter will describe the processes underlying the capability of the genus Bacillus spores to survive under extreme conditions.
7. Initiation of Germination in Bacillus and Clostridium Spores
Dormant bacterial spores of the genera Bacillus and Clostridium employ receptors belonging to the GerA family as environmental sensors. The recognition and binding of appropriate small molecule ligands, termed germinants, irreversibly commits the spore to initiate the cascade of biophysical and hydrolytic reactions that comprise the germination process. Germination is a crucial event in the lifecycle of spore forming organisms, since in virtually all cases the detrimental properties associated with the bacterium are manifested during vegetative growth. However, despite intensive research spanning several decades, few details concerning the precise function of the nutrient germinant receptors (other than their ability to trigger spore germination in response to specific germinants) or the molecular mechanisms that underpin the earliest stages of the process, are known. The current chapter describes genetic, structural and functional aspects of spore germinant receptors, highlighting diverse functional roles and mechanistic strategies associated with GerA-type receptors. Indeed, present outputs from mutational and bioinformatic analyses seem to support the notion that, whereas the general mechanism of triggering germination may be largely conserved between Bacillus and Clostridium species, a number of different mechanisms involving GerA-type receptor proteins, and perhaps unrelated proteins, also exist, and are employed by at least by some pathogenic Clostridium.
8. Germination of Bacillus anthracis Spores
Jonathan D. Giebel, Katherine A. Carr and Philip C. Hanna
Bacillus anthracis exists in two morphologically distinct states, the spore and the vegetative cell. B. anthracis spores, not the vegetative bacilli, are the etiologic agent of the disease anthrax. Anthrax infections result from these inert spores entering a suitable host, recognizing that the environment is suitable for growth, and then transitioning to the rapidly growing, toxin- and capsule-producing vegetative cell through a programed germination process whose proteins are produced during spore formation and remain inactive until triggered by specific cues. The process is, generally, conserved among spore forming bacteria. In this chapter we outline some of the unique attributes of B. anthracis spore germination, such as the spatial and temporal regulation of spore germination inside a host, the repertoire of small molecule germinants recognized by the spore, their concomitant germinant receptors and the group of lytic enzymes necessary to complete the cortex degradation that allows for vegetative cell outgrowth and discuss how the proper functioning of each of these steps contributes directly to the pathogenesis of this organism.
9. Degradation of Spore Peptidoglycan During Germination
David L. Popham, Jared D. Heffron and Emily A. Lambert
During spore germination, several spore components are broken down and are discarded or recycled. The first major degradative step in germination is depolymerization of the spore cortex peptidoglycan. This is essential to allow full hydration of the spore core and the resumption of cellular metabolism. The spore cortex is a thick layer of peptidoglycan with structural modifications that differentiate it from vegetative cell wall material. Germination-specific cortex lytic enzymes exhibit specificity for the muramic-δ-lactam modification of the peptidoglycan strands. As no protein synthesis can take place during germination prior to cortex breakdown, the cortex lytic enzymes must be produced during spore formation and packaged within the dormant spore in an inactive and highly stable form. A mechanism(s) must then exist for the activation of lytic enzymes during germination. This chapter will cover the current knowledge concerning the identities of cortex lytic enzymes in Bacilli and Clostridia, their expression and incorporation into dormant spores, the mechanisms that hold them inactive during spore dormancy and their activation during germination, and their specific lytic activities, substrates, and products.
10. Water and Cations Flux During Sporulation and Germination
Daniela Bassi, Fabrizio Cappa and Pier Sandro Cocconcelli
Bacterial spore water uptake and solute permeability are hypothesised to play a central role during the sporulation and germination processes but the leading mechanisms are at now poorly understood. In this chapter, a state of the art and the last experiments to better study this topic are presented. In the first part, major questions are addressed to the role of water content in spore dormancy and resistance, to the mechanisms which lead to spore dehydration and to the permeability of the spore layers to water and solutes. In the second part, the attention is focused on the ion composition of spores and their importance for the spore life cycle, with particular attention to the leading mechanisms acting during spore formation in the mother cell and during the germination process and outgrowth.
11. The Spore as an Infectious Agent: Anthrax Disease as a Paradigm
James M. Vergis, Christy L. Ventura, Louise D. Teel and Alison D. O'Brien
The zoonotic disease anthrax is caused by the inhalation, ingestion, or cutaneous inoculation of Bacillus anthracis spores into a susceptible host. While inhalational anthrax is rare in humans, the 2001 distribution of B. anthracis spores through the U.S. mail led to an acceleration of research on disease progression in animal models, vaccines and therapeutics. After inoculation, spores are rapidly phagocytosed by macrophages and dendritic cells where they germinate into vegetative bacilli that produce toxins and capsule, both of which facilitate dissemination in the host. The vegetative bacilli are susceptible to antibiotics, the only licensed post-exposure treatment against B. anthracis. Despite the fact that the spore is the infectious particle, the currently approved vaccines for humans in the US and UK are derived from the vegetative culture filtrate of a toxigenic, nonencapsulated B. anthracis strain; the vaccines primarily contain Protective Antigen (PA). Here we describe the role of the spore in anthrax disease progression. In addition, we discuss PA-based vaccines and therapeutics, as well as historical applications of spore-based vaccines. Finally, we present current research focused on spore antigen vaccines and germination modulators.
12. Heterogeneity in Bacterial Spore Populations
Peter Setlow, Jintao Liu and James R. Faeder
Genetically identical populations of spores of Bacillus and Clostridium species invariably exhibit significant heterogeneity, including wide variation in individual spores' resistance to wet heat, speed of germination, and requirements for heat activation before germination. This heterogeneity appears due to both stochasticity in gene expression and variations in culture conditions during sporulation, and presumably facilitates the survival of spore populations. However, small percentages of individuals in spore populations that germinate extremely slowly (superdormant spores) or are much more resistant to environmental insults greatly complicates spore eradication in the food and health care industries. Recently a number of methods have been developed to simultaneously analyze the behavior and biochemical properties of multiple individual spores in populations, in particular their: i) response to wet heat; ii) germination; iii) levels of specific components including dipicolinic acid and individual proteins; and iv) protein status, whether native or denatured. Results using these methods have led to a much greater appreciation of the scope of spore heterogeneity. In addition, analysis of the germination of multiple individual spores has facilitated the development of a quantitative model for spore germination, and such a model suggests experiments to further elucidate the mechanism(s) of spore germination.
13. Detection of Bacterial Spores: Prospects and Challenges
Sainath Rao Shilpakala, Krishna Mohan V. Ketha and Chintamani D. Atreya
Bacteria of several genera are able to form endospores when subjected to certain starvation conditions. The endospores are dormant forms that are structurally and biochemically different from the corresponding growing or vegetative cells. These bacterial endospores resist antibiotics, desciccation, and ordinary boiling than the vegetative cells. The detection of bacterial endospores can be important for a wide variety of purposes. In the sanitation and hygiene fields, detection of bacterial spores can be critical to monitor indoor environments, water quality and food quality. Similarly, in the public health perspective detection of bacterial spores is very important in ensuring regulation of safer transfusion and other therapeutic products, administered either orally or intravenously. Monitoring of bacterial spores is gaining importance even in other areas such as agriculture wherein soil or plant samples are periodically monitored for bacterial spores to ensure high levels of the bacterial population/toxin to be effective against insect pests. More recently, with the possibility of bacterial spores being used as bio-threat agents there has been an augmented effort in developing much more sensitive, specific and rapid detection systems for bacterial spores. The review discusses about current methods of bacterial spore detection and the challenges involved.
14. Properties and Detection Methods of Bacilli Spores in Food and in Medical Settings
Olga Tarasenko, Pierre Alusta, Sergey Kazakov and Kalle Levon
Numerous members of Bacillus and Clostridium genera are widely distributed in nature. Their control is considerably important since these species are responsible for a variety of food spoilage, foodborne illnesses, and food-poisoning problems, in addition to their potential use as biological warfare agents. The present study concentrates on the taxonomic Bacillus cereus group covering studies of genetics and morphology, including that of harmful bacteria. Moreover, several classical issues in spore resistance biology are being considered. Examining a number of reports will provide us with morphological properties of spores as well as their biochemical composition. We will then extrapolate on some of the findings, thus enabling us to determine the role of how to identify proteins in spore immunology and outline a possible issue in this regard. This paper presents an overview of different physicochemical techniques meaningful for spore detection and identification as a basis for biosensor construction and/or other detection techniques. Special attention is given to different methods of spore detection and identification including but not limited to microbiological, immunological, genetic, and radiometric methods, automation of assay procedures, biosensors as well as new approaches. Future directions for development and problems related to biosensors are discussed in the final part of this review.
15. Expression of Recombinant Proteins Using Bacillus subtilis Spores
Luis Carlos de Souza Ferreia and Wolfgang Schumann
Surface display of peptides and proteins on recombinant phages and bacterial cells has developed into a powerful technique to identify proteins with desired properties with multiple applications. This technique is of utmost importance as a tool for fundamental and applied research in the fields of microbiology, biotechnology and vaccination. Another promising bioparticle are spores produced by Bacillus subtilis cells that allow fusion of recombinant proteins to one of several outer spore coat proteins. There are several reports were antigens and proteins of biotechnological interest have been successfully displayed on the outside of spores. A major advantage of spores over bacterial cells is their intracellular biogenesis that does not require specific secretion sequences of the cellular apparatus. Therefore, heterologous proteins do not have to cross membranes and correct folding of these proteins can be achieved with the aid of cytoplasmic folder chaperones.
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(EAN: 9781908230003 9781912530434 Subjects: [microbiology] [bacteriology] [medical microbiology] [molecular microbiology] )