Bacteriophages: Biology, Technology, Therapy

Preface
Contents
About the Editors
Contributors
Part I: Introduction to Bacteriophages: Biology, Technology, Therapy
	Introduction to Bacteriophages
		Bacteriophages
		On the Nature of Bacteriophages
		Antibiotics: From Savior to Crisis
		Antibiotics: A Biological Approach
		References
Part II: Bacteriophage Biology
	Structure and Function of Bacteriophages
		Introduction
		Overview of Phage Families
			The Leviviridae Family of Single-Stranded RNA Phages
				Assembly
				Structure
			The Microviridae Family of Single-Stranded DNA Phages
				Assembly
				Structure
			The Inoviridae Family of Filamentous, Single-Stranded DNA Phages
				Assembly
				Structure
			The Cystoviridae Family of Double-Stranded RNA Phages
				Assembly
				Structure
		Overview of Bacteriophages Containing Double-Stranded DNA Genomes
			The Tectiviridae Family
				Assembly
				Structure
			The Corticoviridae Family
				Assembly
				Structure
		Overview of Order Caudovirales
			Caudovirales Head Assembly
			Caudovirales Head Structure
			Podoviridae Tail Assembly and Structure
			Siphoviridae Tail Assembly
			Siphoviridae Tail Structure
			Myoviridae Tail Assembly
			Myoviridae Tail Structure
		Conclusions and Perspectives
		Cross-References
		References
	Adsorption: Phage Acquisition of Bacteria
		Introduction
		Free Virions to Virocells
		Movement of Free Phages
			Virion Diffusion
			Non-diffusive Movement
		Encounter
		Attachment
			Overview
			Adsorption Factors and Reversible Attachment
			Irreversible Attachment
		Adsorption Kinetics
			Importance of Phage Titers
			Adsorption Rate Constants
			How Fast Adsorption Is Fast Enough?
			Adsorption Rate Constant Determination
		Genome Translocation
		Conclusion
		Cross-References
		References
	Temperate Phages, Prophages, and Lysogeny
		Introduction
		Lysogeny and Lysogenic Cycles
			Stably Associating with Host Bacteria
			Benefiting the Lysogen
			Lysis-Lysogeny Decisions
			Induction
			Immunity
			Lysogenic Conversion and Morons
		Bacteriophage Lambda
			The Lysogeny Decision
		Lambda and Lambdoid Genomes
		Prophages P2 and P2-Likes
			P2 Virion and Genome
			P2 Lysogeny
		Temperate Phages of Staphylococci and Streptococci
			Staphylococcal Phages
				Morphological Families and Classification
			Streptococcal Phages
				Morphological Families and Classification
		Role of Prophages in Modulation of Bacterial Host Biology and Evolution
			Toxins and Virulence Factors Encoded by Prophages
				Staphylococcus aureus
				Clostridium botulinum
				Vibrio cholerae
				Corynebacterium diphtheriae
				Streptococcus pyogenes
		Conclusions
		References
	Bacteriophage-Mediated Horizontal Gene Transfer: Transduction
		Introduction
		Overview of Transduction
		Historical Perspectives on Horizontal Gene Transfer in Bacteria
			The Discovery of Transformation
			The Discovery of Conjugation
			The Discovery of Transduction
		Examples of Transducing Phages
			Bacteriophage P1
			Bacteriophage P22
			Lambda (λ)
			Mu
		Phage Features Important for Transduction
			Host DNA Degradation
			DNA Packaging Method
			Mobile Genetic Elements Within Phage Genomes
			Integration Site Specificity
		Examples of Transduction Hijackers
			Gene Transfer Agents
			Phage-Inducible Chromosomal Islands
		Methods to Detect and Characterize Transducing Particles
			Phenotypic Changes of the Bacterial Host
			Phenotypic Changes of the Transducing Particle
			Genotypic Changes of the Bacterial Host
			Genotypic Changes of the Transducing Particle
		Potential Risks to Human Health Posed by Transduction
			Transduction and the Spread of Antibiotic Resistance Genes
			Transduction and the Implications to Phage Therapy
		Conclusions
		Cross-References
		References
	Genetics and Genomics of Bacteriophages
		Introduction
		The Nature of Bacteriophage Genomes: Evolution and Mosaicism
		Main Structural Features
			Structural Components of Tailed Bacteriophages
			Structural Components of PFP Bacteriophages
			Bacteriophage T4
		Hallmark Genes and Variability
			Hallmark Genes and Bacteriophage Evolution
		Comparative Genomics
			Bacterial Comparative Genomics: The Impact of Bacteriophages
			Comparative Genomics of Bacteriophages: Phylogeny and Evolution
			Comparative Genomics and Bacteriophage Taxonomy
			Comparative Metagenomics
		Functional Genomics
			Functional Genomics and Genetic Characterization
			Site-Directed Mutagenesis (SDM) and Gene Knockouts
		Transcriptomics
			Host Response to Bacteriophage Infection
		Discussion and Future Perspectives
		References
	Bacteriophage Discovery and Genomics
		Introduction
		Phage Discovery and Genomics as a Platform for Science Education
			The Phage Hunters Integrating Research and Education (PHIRE) Program
			The Mycobacterial Genetics Course
			The Science Education Alliance Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) Program
		A View of Bacteriophage Genetic Diversity
		Concluding Remarks
		Cross-References
		References
	Bacteria-Phage Antagonistic Coevolution and the Implications for Phage Therapy
		The Coevolutionary Process and the Red Queen´s Race
		Experimental Studies of Bacteria-Bacteriophage Antagonistic Coevolution
		Mechanisms of Bacterial Resistance to Lytic Bacteriophage and Phage Counter-Resistance
		Effects of Resistance and Counter-Resistance on Fitness and Phenotype
		(Co)Evolutionary Considerations of Phage Therapy
		Cross-References
		References
	Bacteriophage Ecology
		Introduction
		Phage Presence in Nature
			Viable Phage Counts
			Phage Total Counts
			Nucleic Acid Sequence-Based Determinations
				Linking Phages to Bacteria via Metagenomics
			Biogeography
				Phage Prevalence Within Environments
				Phage Movement
				Phage Distributions Within Environments
		Phage Population Ecology
			Phage Life Cycles as Ecological Phenomena
			Virion Durability
			Virion Attachment Affinity
			Evasion of Host Defenses
			Phage Latent Periods
			Phage Burst Size
		Phage Community Ecology
			Predators, Parasites, or Parasitoids?
			Killing the Winner
			Phage Exploitation of Biofilm-Associated Bacteria
		Phage Ecosystem Ecology
		Summary
		Cross-References
		References
	Bacteriophage Pharmacology and Immunology
		Introduction
		Safety Considerations in Phage Choice for Phage Therapy
			Avoiding Temperate Phages
			Avoiding Phages Encoding Virulence Factors
			Avoiding Transducing Phages
			Summary: Safety Considerations
		Phage Therapy Pharmacology Basics
			Pharmacokinetics and Pharmacodynamics
			Pharmacokinetics in More Detail
			Conceptualizing Pharmacology
		Passive Treatment Versus Active Treatment
			Passive Phage Therapy
			Active Penetration
			Active Phage Therapy
			Summarizing Phage Therapy Pharmacological Phenomena
		Phage Interactions with Immune Systems
			Overview of Immunity
			Adaptive Immunity
			Innate Immunity
		Effects of Immune Response on Phage Pharmacokinetics
		Relation of Immune Response to Phage Pharmacodynamics
		Conclusion
		Cross-References
		References
	Phage Infection and Lysis
		Introduction
		Productive, Reductive, and Destructive Phage Infections
			Phage-Productive Infections
			Phage-Reductive Infections
			Phage-Destructive Infections
			Phage Growth Parameters
				Latent Periods
				Burst Sizes
				Utility for Phage Therapy
			Strictly Lytic Phages for Phage Therapy
		Infection
			The Eclipse
			Phage Gene Expression
			Phage Genomes and Replication
			The Latent Period Continues. . .
			. . .And Continues?
			The Rise
			Host Physiology Considerations
		Virion Release
			Holin-Mediated Lysis from Within
				Lysis-Mediating Phage Proteins
					Antiholins
			Inhibition of Peptidoglycan Production
			Chronic Release
		Determination of Phage Growth Parameter Values
			Virion Durability Determination
			Eclipse, Latent Period, Burst Size, and Rise
			Phage Population Growth Rates
			Continuous Culture
			Plaques
		Conclusion
		Cross-References
		References
Part III: History of Bacteriophages
	The Discovery of Bacteriophages and the Historical Context
		Discovery in Science: No Eureka Moment
		Microbes Before Twort and d´Herelle
			Viruses
			Filters
			``Fact Making´´: Bacterial Lysis and ``Lytic Principles´´
			Cultures, Cells, Microbial Mutations
		Twort and d´Herelle: The Canonical Account
			The Canonical History of Bacteriophage
		Becoming Phage: 1920-1940
		Phage Discovery Completed
		Discovery and Priority Revisited
		Cross-References
		References
	Early Therapeutic and Prophylactic Uses of Bacteriophages
		Introduction: A Brief History
		Phage Therapy for Wound Treatment, Surgery, and Dermatology
		Phage Therapy for Treatment of Enteric Infections
		Prophylactic Use of Phages
		Intravenous Staphylococcal Bacteriophage: The Highest Achievement of the Georgian Scientists
		Conclusions
		References
Part IV: Bacteriophage Technology
	Isolation of Bacteriophages
		Glossary
		Introduction
		Basic Phage Isolation Techniques
			Biases in Isolation
			Spot Testing
			Increasing Phage Concentrations
			Generation of Pure Isolates and Phage Stocks
			Storing Isolated Phages
		Where to Hunt for Phages
			Isolation from the Environment
			Common Sources for Isolation of Therapeutic Phages
			Isolation from Lysogens
			Isolation from Diverse Environments
		Resulting Phage Phenotypes
			Altered Procedures and Their Effect on Host Range
			Plaque Morphology Indicates Diverse Phage Characteristics
		Phage Isolation for Biotechnology
			Implications for the Use of Phages as Antimicrobials
			High-Throughput (HTP) Phage Isolation
		Conclusions
		Cross-References
		References
	Bacteriophage Use in Molecular Biology and Biotechnology
		Early Contributions to Molecular Biology
			Early Studies of Phage Chemical Composition and Physical Structure
			Spontaneous Mutation and Heredity
		Phage Growth and Plaque Formation
			Phage Typing
			Phage Amplification and MALDI-TOF MS for Bacterial Identification
		The Discovery of Lysogeny and Prophages
			The First Unequivocal Demonstration of Lysogeny
			The Discovery of Phage λ and the Beginning of Prophage Genome Mapping
			The Discovery of Phage P1 Used in Bacterial Transduction
		The Discovery of Site-Specific Recombination and Its Applications
			The Campbell Model of Phage Integration
			The Determination of Attachment Sites on Host and Phage DNA Molecules
			The Development of Site-Specific Integrating Plasmids
			Discovery of the Cre/LoxP System in Phage P1
			Applications of the Cre/Lox System in Genetically Engineered Mice
		Discovery of Bacterial DNA Restriction and Modification Systems
			Discovery of the Restriction/Modification Phenomenon
			Discovery of the Role of Methylation in Restriction/Modification
			The First Identification of a Specific Restriction Site in Bacterial DNA
			The Development of Molecular Cloning
			The Development of Restriction Mapping
		The Discovery of Viral Metabolic Products
			The Discovery of Thymineless Death and Its Implications for Chemotherapy Research
		The Role of Phages in Understanding Gene Structure, Expression, and Regulation
			Luria and Delbruck´s ``Fluctuation Test´´ Shows That Spontaneous Mutations Occur in Bacterial Genes
			Hershey and Chase Demonstrate That DNA Is the Hereditary Material
			Benzer Maps the Fine Structure of Genes in the Phage T4 rII Region
			The Discovery that Messenger RNA Is a Direct Product of Genes
			The Determination of the Genetic Code
			The First Observation of Gene Regulation Due to a Repressor Factor (The PaJaMo Experiment)
			The First Isolation and Characterization of a Repressor Factor Involved in Gene Regulation
			Isolation of the ρ Termination Factor and Characterization of Termination/Antitermination in λ
			The Construction of a Synthetic Genetic Regulatory Circuit that Exhibits Complex Behavior
			Refactoring Phages to Learn About Complex Genetic Regulatory Circuits at the Organismal Level
		Conclusion
			Systems Biology
			Evolution
			Ecology
		Cross-References
		References
	Detection of Bacteriophages: Phage Plaques
		Introduction
		Plaques
			Initiating Plaques
			Bacterial Lawns, Plaques, and Spots
			Plaque-Forming Units
			Too Many or Too Few Plaques
		Plaques Versus Spots
		Plaque Formation
			Phage Clumped Dispersion
			Bacterial Clumped Dispersion
		Plaque Size
		Plaque-Based Phage Characterization
			Efficiency of Plating
			Efficiency of Center of Infection
			Mixed-Indicator Technique
		Conclusions
		References
	Detection of Bacteriophages: Statistical Aspects of Plaque Assay
		Introduction
		Overview
		TFTC and TNTC
		Spot Counts Versus Plate Counts
		Number of Repeats
		Utility of Trimmed Means
		Number of Dilution Series
		Utility of Larger Volumes
		When Comparisons Matter
		Conclusions
		Cross-References
		References
	Detection of Bacteriophages: Electron Microscopy and Visualization
		Introduction
		Electron Microscopy
		Initial Electron Microscopy of Phages
		Staining
		Metal Shadowing
		Negative Staining
		Thin-Sectioning, Room-Temperature and Cryogenic
		Frozen-Hydrated Specimens, Unstained and Negatively Stained
		Scanning Electron and Helium-Ion Microscopy
		Scanning Transmission Electron Microscopy, Dark-Field
		In Situ, Liquid-Cell Electron Microscopy
		Analytical Electron Microscopy
		Two-Dimensional Image Averaging and Diffraction
		Three Dimensions from Two
		Immuno-EM
		Conclusion
		References
	Detection of Bacteriophages: Sequence-Based Systems
		Introduction
		Overview of Sequencing Methods
			Short-Read Platforms
			Long-Read Platforms
			A Note on Sequence Data and Sharing
		Applications of Sequencing-Based Detection Methods
			Gene-Based Detection of Bacteriophages
				Cyanophages
				T4-Like Phages
				T7-Like Phages
				Gokushoviruses
				Other Potential Signature Genes
			Genome Sequencing
			Sequence-Based Identification of Prophages
			Metagenomics-Based Detection of Bacteriophages
		Conclusions
		Cross-References
		References
	Novel Approaches for Detection of Bacteriophage
		Mass Spectrometry
		Raman Spectroscopy
		Summary
		Cross-References
		References
	Bacteriophages in Nanotechnology: History and Future
		Introduction
		Bacteriophage Capsids
		Phage Virions and Phage Proteins in Biosensors
			Attachment of Whole, Non-modified Virions
			Attachment of Phage Proteins
			Signal Transduction Methods
		Phage Display
		Phages in Nanostructured Materials
			Applications Using Whole Phages
			Applications Using Phage Display-Generated Peptides or Proteins
			Whole Phage in Structured Materials
		Phage Catalysts
		Phages as Drug Delivery Carriers
		Phage-Containing Antibacterial Materials
		Challenges to Commercialization
		Conclusion
		Glossary
		References
	The Selection and Optimization of Phage Hosts
		Introduction
		Phage Propagation
		Surrogate Hosts
		Prophages
		Reducing Host Toxicity
		Endotoxin
		Conclusion
		References
	Bacteriophage Manufacturing: From Early Twentieth-Century Processes to Current GMP
		What Is GMP, and When Is It Needed?
			Definition of GMP
			History of GMP Guidelines, Need, Definition, and Place of Bacteriophage
			Evolution of GMP: Current Guidelines and Discrepancies Between Regions
		History of Phage Production. How It Was Handled Until Now
		Applying GMP to Bacteriophages
			General Considerations
			Cell Culture Media AOF, No Animal, or Human-Origin Material
			Biological Material Choice
			Phage Banking
			Cell Banking
		Manufacturing
			Why Prefer Single Use to Stainless Steel?
			Upstream Process: Phage Production
			Downstream Process: Phage Purification
			Formulation, Fill, and Finish
		Control Quality
			Cell Substrate Quality Control
			Bacteriophage QC
				Phage Host Range
				Host Cell Protein
				Endotoxins and Pyrogens
				Transmission Electron Microscopy
				Genome Sequence
				RFLP (Restriction Fragment Length Polymorphism)
				RAPD (Random Amplified Polymorphic DNA)
				Phage DS Infectious Titer
				Stability Studies
			Future of the Regulation for Phage-Based Drug Products
		Perspectives of Phage Manufacturing
			Impact of GMO Phage Manufacturing
			Perspective About Phage Quality Control
			Conclusion
		Cross-References
		Glossary
		References
	Intellectual Property Issues for Bacteriophages
		Patents and Patenting
			What Is a Patent?
			Why Apply for a Patent?
			Patentability and Freedom-to-Operate Assessments
			What Is Needed to Apply for a Patent?
			Novelty, Inventive Step, and the Undue Burden
			Unity of the Invention
			Elements of a Typical Patent Application
			From Filing to Grant
			After Award
			Timing and Patent Terms
			First to File Versus First to Invent
			Safe Harbors and Extinct Submarines
			Future Developments
		Patenting of Bacteriophages
			Patenting Natural (Unmodified) Bacteriophages
			Patenting Genetically Modified Bacteriophages
			Patenting Novel Processes or Technologies
			Bacteriophage Patents
			Accessing Bacteriophage Patents Online
		Glossary
		References
	Bacteriophage as Biocontrol Agents
		Introduction
		Biological Control
		Bacteriophages in Nontherapeutic Biological Control
		Efficacy of Biological Control
		The Damage Threshold: A Biocontrol Concept
		IPM
		The Kinetics of Biological Control
		Implications for Clinical Trials
		Nature of the Agent
		Summary
		Cross-References
		Glossary
		References
Part V: Agriculture, Food, and Environmental Use of Bacteriophages
	Bacteriophages as Bio-sanitizers in Food Production and Healthcare Settings
		Introduction
		Phages as Bio-sanitizers on Food Contact Surfaces
			Phages as Bio-sanitizers for Listeria monocytogenes
			Phages as Bio-sanitizers for Other Foodborne Pathogens
				Escherichia Coli O157:H7
				Salmonella spp.
		Phages as Bio-sanitizers in Hospital Settings
		Phage Components as Bio-sanitizers or Sanitizing Adjuncts
		Phage Application Approaches
		Conclusion and Future Directions
		References
	Biofilm Applications of Bacteriophages
		Applications of Bacteriophages to Biofilms and Surface-Attached Bacteria
			Introduction
		Biofilms in Food Industrial Settings
			Prevalence of Biofilms
			Biofilm Formation on Surfaces
			Impact of Biofilms in Different Food Industry Settings
				Dairy Industry
				Poultry Industry
				Red Meat Industry
				Seafood Industry
				Fresh Fruit and Vegetable Industry
			Currently Used Removal Strategies
		Bacteriophage Application to Biofilms and Other Surface-Attached Bacteria Formed in the Food Industry
			Application of Bacteriophages to Equipment Surfaces and to Working Surfaces and to Enhance Equipment Performance
			Application of Bacteriophages to Foods
				Fresh-Cut Fruits and Vegetables
				Red and Poultry Meats
				Dairy Products
		Single Bacteriophage Versus Cocktail Approach and Other Approaches
		Challenges that Bacteriophages Face in Industrial Environments
		Bacteriophages and Biofilms in the Clinical Setting
			Background
		Bacteriophage-Biofilm Interactions
		Applications
		Conclusion
		Cross-References
		References
	Industrial Processes Involving Bacteriophages
		Introduction
		Highly Complex Bacterial Culture-Driven Bioprocesses
		Bioprocesses Driven by Well Defined, Mixed Cultures
		Monocultures in Industrial Biotechnology: Pharmaceutical Catalysts, Bacterial Biomass, and Green Chemistry
		Pharmaceutical Bioprocesses
		Phage Growth in Bioprocesses
		Prophages in Bioprocesses
		Bacteriophage Contamination Troubleshooting and Prevention
		Bacteriophages as Antibacterials in Bioprocesses
		Conclusions
		Cross-References
		References
	Crop Use of Bacteriophages
		Management of Plant Bacterial Diseases
		Use of Bacteriophages in Agriculture
		Factors that Challenge Use of Phages
		Bacteriophages and the Bacterial Carrier System
			Phage as Infectious Bacteriostatic Particles
			Application Challenges in Agriculture
			Historical Perspective on the Phage Delivery System
			Avoidance of Host Resistance and Other Considerations
			Field Application of Carrier-Phage
		Cross-References
		References
	Food Safety
		Introduction
		Food and Microorganisms
			Limiting Food Contamination
			Food Legislation Enhances Safety
			Trends Towards Industrialized Convenience Foods
		Bacteriophages and Food Safety
		Bacteriophages for the Biological Control of Gram-Negative Bacteria
			Treatment of Poultry by Campylobacteriophage
			Control of Salmonella enterica Using Bacteriophage
			Bacteriophage for the Control of Escherichia coli
			Phages Infecting Yersinia enterocolitica
			Cronobacter sakazakii
			Pseudomonas aeruginosa
			Legionella pneumophila
			Vibrio parahaemolyticus
		Control of Gram-Positive Bacteria by Bacteriophage Treatment
			Listeriophages Are Well Suited for Biocontrol
			Bacteriophages for the Control of Staphylococcus aureus
			Control of Bacillus cereus in Food
		Future Perspectives
		Conclusion
		Glossary
		References
	Bacteriophage Utilization in Animal Hygiene
		Introduction
		Brief History of Phage Use in Animal Hygiene
		Live Animal Pathogen Reduction Treatments
			Phages as a Strategy to Improve Hygiene in Live Animals
			Challenges to Phage Treatment of Live Animals
		Poultry Specific Issues and Phage Usage
			Poultry Health
		Swine Specific Issues and Phage Usage
			Swine Health
		Ruminant Specific Issues and Phage Usage
			Hide-Specific Spray Treatments for Cattle
			Cattle Health
		Conclusions and the Future
		Cross-References
		References
Part VI: Therapeutic Use of Bacteriophages
	Current Updates from the Long-Standing Phage Research Centers in Georgia, Poland, and Russia
		Introduction
		Current Status of Georgian Research
			History
			Spin-Off Entities
			International Collaboration
			Targeting Antibiotic-Resistant Bacteria
			Treatment of Cystic Fibrosis-Associated Infections
			Treatment of Urinary Tract Infections
			Targeting Infected Wounds
			Conclusions: Georgian Phage Therapy
		Current Status of Polish Research
			The Hirszfeld Institute
			Progress of the Phage Therapy Unit 2012-2017
			Immunological Responses to Phages
			The Hirszfeld Institute´s Phage Collection
			Plasmid-Prophage-Free Hosts for Optimal Therapeutic Phage Preparations
			Our Published Output
			Conclusions: Polish Phage Therapy
			Acknowledgments: Polish Phage Therapy
		Current Status of Russian Research
			Background
			Russian Phage Therapy Research and Implementation
			Conclusions: Russian Phage Therapy
		References
	The Use of Bacteriophages in Veterinary Therapy
		Introduction
		History of Bacteriophages in Veterinary Use
		Systemic Escherichia coli Infections
		Systemic Salmonella Infections
		Intestinal E. coli Infections
		Intestinal Salmonella Infections
		Intestinal Campylobacter Infections
		Other Systemic Infections
		Other Infections
		Use in the Food Production Chain
		Applications, Potential, and Problems
		Regulatory Approval
		Cross-References
		References
	Enzybiotics: Endolysins and Bacteriocins
		Endolysins, the Model Enzybiotic
			Lysis from Without
			Endolysin Structure
			Bacteriolytic Mechanism
		Antimicrobial Development of Endolysins
			Specificity and Resistance
			Toxicity
			Immunogenicity
			Synergy
		Endolysin Applications
			Medicine
			Food Safety
			Agriculture
		Endolysin Engineering Approaches
			Chimeragenesis
			Directed Evolution
			Structure-Guided Site-Directed Mutagenesis
			Gram-Negative Engineering Approaches
			Targeting Intracellular Pathogens
		Regulatory Aspects of Endolysins
		Other Phage-Based Enzybiotics
			Particulate Bacteriocins
			Applications in Crop Health
			Applications in Medicine
			Engineering Particulate Bacteriocins
		Concluding Remarks
		Cross-References
		References
	Phage Display Technology and the Development of Phage-Based Vaccines
		Phage Display Technology
			Introduction
			Combinatorial Chemistry and Display Phage Technology
				Combinatorial Chemistry
				Phage Display Technology
				Display Phage
				Affinity Selection
				Amplification Bias
				Subtractive Panning
				Biopanning Apparatus
			Characterization of Selected Ligands
				Phage Capture Assay
				Western Blot Analysis
				Inhibition ELISA
				DNA Sequence Analysis
			Conclusion
		Phage as Delivery Vehicles
			Introduction
			Preparation of Modified Phages
				Modification of Coat Proteins
				Modification and Addition of Nonstructural Genes
				Synthetic Phages
			Applications of Modified Phages
				Phages as Vaccine Delivery Vehicles
				Phage-Mediated Protein Vaccines
				Phage DNA Vaccines
				Phages as Gene Therapy Vectors
			Conclusion
		Development of Monoclonal Antibody Mimics
			Introduction
			Small Peptide Mimics of mAbs
				Mimotopes
			Selection of mAb Mimics (Anti-epitopic Peptides)
			Peptide-Based Cytotoxicity
			Conclusion
		References
	Phage Therapy Collaboration and Compassionate Use
		Introduction
		Regulation of Compassionate Phage Therapy
		General Process of Compassionate Phage Therapy
			Establishing Medical Need and Complying with Regulations
			Identifying Sources of Phages
			Phage Susceptibility Testing
			Preparing Phages for the Patient
			Administering Phages to the Patient
			Coordinating Compassionate Phage Therapy
			Costs of Compassionate Phage Therapy
		Recent Examples of Compassionate Phage Therapy
			Diabetic Toe Ulcers (US/Georgia Collaboration)
			Disseminated Acinetobacter Infection (US Collaboration)
			Posttransplant Burkholderia Infection (US/Canada Collaboration)
			Other US Cases
			Polymicrobial Bone Infection (Israel/US Collaboration)
			Engineered Phage Therapy (UK/US Collaboration)
			Global Crowdsourcing of Klebsiella Phages (Finland/US/Israel/UK/Switzerland Collaboration)
		New Systems Created to Facilitate Compassionate Phage Therapy
			Phage Therapy at the Queen Astrid Military Hospital
			Phage Directory
			Center for Innovative Phage Applications and Therapeutics (IPATH)
			Vitalis Phage Therapy
		Summary and Concluding Remarks
		Cross-References
		References
	Clinical Trials of Bacteriophage Therapeutics
		Introduction
			From Antibiotics to Phage
			From East to West
		Modern Phage Trials to Date
			P. areuginosa Chronic Otitis Externa
			Chronic Venous Leg Ulcers
			Immunoresponse in HIV Patients
			E. coli Pediatric Diarrhea: The Bangladesh Trials
			Burn Wound Infections
			S. aureus Diabetic Toe Ulcers
			S. aureus Chronic Rhinosinusitis
		Case Reports
			Phage Therapy Unit, Wroclaw, Poland
			PhagoBioDerm
			The Eliava Institute and Medical Tourism
		Hurdles to the Development of Phage Therapy in the Western World
			Regulatory Framework
			Finances
			Resistance
			Biological Considerations
		Recommendations
		Conclusion
		Cross-References
		References
	Selection of Disease Targets for Phage Therapy
		Introduction
		Current Position
		Issues in the Selection of the Target Disease
			Bacterial Nature of the Underlying Disease
			Unmet Need
			Limited Range of Bacterial Targets
			Bacteria Causing the Disease Are Identified
			Bacteria Targeted Are Responsible for the Observed Pathology
			Availability of Model Systems for Preclinical Work
			Practicality of Clinical Trials
			Accessibility of the Bacterial Target
			The Effect of Bacterial Numbers
		Issues in the Selection of Bacteriophages
			Availability of Bacteriophages
			Suitability of Bacteriophages for Therapeutic Use
			Lysogeny
			Transduction and Toxin Carriage
			Host Range
			Other Issues with Bacteriophage Mixtures
			Storage and Stability
			Commercial Considerations
			Suitability of Product
		Considerations on the Nature of Bacteriophages
		Summary
		Glossary
		References
	Regulatory Considerations for Bacteriophage Therapy Products: USA
		Background
		Regulatory Oversight of Biological Products
		Chemistry, Manufacturing, and Controls for Bacteriophage Products
		Expanded Access to Investigational Drugs for Treatment Use
		Defined Bacteriophage Products Versus Bacteriophage Banks
		Conclusions
		References
	Regulatory Aspects of the Therapeutic Use of Bacteriophages: Europe
		Background
		Regulatory Oversight and Development Guidance
			Regulatory Framework
			Development Guidance
				Quality Parameters
				Non-clinical and Clinical Development
		Phage Therapy Modalities and Ongoing Research Activities
		Access Issues
		Conclusion
		References
Index