Animal Biotechnology: Models in Discovery and Translation

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Animal Biotechnology: Models in Discovery and Translation
Copyright
Dedication
Contents
List of Contributors
Preface
Acknowledgments
Section I: Human diseases: in vivo and in vitro models
1 Drosophila: a model for biotechnologist
	Summary
	What you can expect to know
	Introduction
		Classical aspects of Drosophila melanogaster
			Physical appearance
			Life cycle
			Drosophila development
			Embryogenesis in Drosophila
			Pattern formation in Drosophila
			Homeotic genes in Drosophila
			Drosophila genome
	History
		Historical perspective of Drosophila contributions to biotechnology
	Principle
	Methodology
		Culturing of Drosophila
		Preparation of Drosophila food medium
			Materials required
		Handling of flies
		Fly disposal
		Egg collection
		Dechorination of eggs
		Preparation of DNA for injection
	Protocols
		Protocol for germ-line transformation in Drosophila
			Materials required
			Procedure
	Ethical issues
	Translational significance
	Clinical significance
		Drosophila-based models for understanding human neurodegenerative diseases
		Drosophila as a model for understanding human metabolic disorders
		Drosophila as a model for understanding nephrolithiasis (kidney stones)
		Drosophila-based model for understanding the human immunodeficiency virus pathology
		Drosophila-based therapeutic peptide production
	Turning point
	World Wide Web resources
	Acknowledgments
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
2 Animal models of tuberculosis
	Summary
	Introduction
	Comparative pathology of tuberculosis in humans and animals
		Characteristics of a model for tuberculosis with respect to infection and pathogenesis
	Pathogen diversity: crossing species barriers
	Host diversity: fundamental processes and fine-tuning
	Animal models of tuberculosis: limits and lessons
	Animal models: contributions in tuberculosis vaccine testing
	Various animal models
		Mouse model
		Guinea pig model
		Rabbit model
		Nonhuman primate model
		Cattle model
	Protocols
		Preparing M. tuberculosis inoculum for aerosol exposure
		Aerosol infection of mice using the middlebrook apparatus
		Aerosol infection of guinea pigs using a madison chamber
		Bacteria loading
		Intravenous infection of mice with M. tuberculosis
		Isolation of samples for determining M. tuberculosis load by real-time-PCR
		Determination of bacterial loads in target organs
		Preparation of lungs or other tissues for histology
		Preparation of lung cell suspension
	Ethical issues
	Translational significance
	World wide web resources
	Safety considerations
	References
	Further reading
	Glossary
	Abbreviations
	Long-answer questions
	Short-answer questions
	Answers to short-answer questions
	Yes/no-type questions
	Answers for yes/no type questions
3 Animal models for neurodegenerative disorders
	Summary
	What you can expect to know
	History and methods
		Introduction
		Neurodegenerative diseases
		Amyotrophic lateral sclerosis
		Spinal muscular atrophy
		Spinal and bulbar muscular atrophy
	Principles
		Genetics of amyotrophic lateral sclerosis
			Superoxide dismutase 1-amyotrophic lateral sclerosis
			Amyotrophic lateral sclerosis: genes implicated in RNA metabolism
				TAR-DNA-binding protein 43
				Fused-in sarcoma/translocated in liposarcoma protein
				C9orf72
		Genetics of spinal muscular atrophy
		Genetics of spinal and bulbar muscular atrophy
	Methodology
		Generation of transgenic mice
			Preparation and purification of transgenic construct (step 1)
			Harvesting donor eggs (step 2)
			Microinjection of transgene to fertilized egg (step 3)
			Implantation of microinjected egg to pseudopregnant female mice (step 4)
			Screening of founder mice for expression of transgene (step 5)
			Establishing stable transgenic line (step 6)
		Cre-loxP technology
		Amyotrophic lateral sclerosis models
			SOD1G37R transgenic mice
			SOD1G93A transgenic mice
			SOD1WT transgenic mice
		Spinal muscular atrophy models
			Severe spinal muscular atrophy mice (mSMN−/−;SMN2+/+)
			Spinal muscular atrophy type II mice (mSMN−/−;SMN2+/+;SMNΔ7+/+)
		Spinal and bulbar muscular atrophy models
			AR-97Q and AR-24Q transgenic mice
	Examples and their applications
		Superoxide dismutase 1-linked amyotrophic lateral sclerosis
			Gain of toxicity from mutant superoxide dismutase 1 established as pathomechanisms through engineering mutant superoxide di...
			Evaluating phenotype and clinical course of mutant superoxide dismutase 1 transgenic mice
			Toxicity from misfolded mutant superoxide dismutase 1 protein
			Non-cell autonomous neurodegeneration demonstrated by superoxide dismutase 1 mouse models
			Stem cell-derived motor neurons established from mutant superoxide dismutase 1 mice
		Other amyotrophic lateral sclerosis
		Spinal muscular atrophy
			Human SMN2 transgenic mice
			Neuron-specific deletion of survival of motor neuron in mice using Cre-loxP systems
		Spinal and bulbar muscular atrophy
			AR-97Q mice as spinal and bulbar muscular atrophy Model
			Androgen hormone and mutant androgen receptor central to spinal and bulbar muscular atrophy pathogenesis
	Clinical correlations
	Protocols
	Ethical issues
	Translational significance
	World Wide Web resources
	Acknowledgment
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers for yes/no type questions
4 Epigenetics and animal models: applications in cancer control and treatment
	Summary
	What you expect to know
	Introduction
	History
	Principle
		Use of mouse models in the epigenetics of cancer
		Examples with applications
			Brain cancer
			Breast cancer
			Colorectal cancer
			Esophageal cancer
			Gastric cancer
			Head and neck cancer
			Lung cancer
			Lymphoma and leukemia
			Prostate cancer
			Liver cancer
			Other approaches
	Methodology
		Methylation profiling
		Histone profiling
		Nucleosome mapping
	Protocols
	Ethical issues
	Translation significance
		Clinical significance
		Web resources
		Turning point
		Flow chart
	World Wide Web resources
	Something interesting about this chapter
	References
	Glossary (terms used in text with examples)
	Abbreviations
	Long answer questions
	Answers to long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
5 Development of mouse models for cancer research
	Summary
	What you can expect to know
	Introduction
	History
	Principle
		Institutional Animal Care and Use Committee approval
		Institutional Animal Care and Use Committee guidelines
	Methodology
		Inbred mice
	Examples with applications
		Immunocompetent mice
			Spontaneous tumor models
			The genetically engineered mouse models
			The Cre/Lox system: a superior genetically engineered mouse model
		Immunodeficient mice
			Allograft transplants
			Xenograft transplants
			Humanized mice
	Checklist for a successful in vivo experiment
	Protocols
		An orthotopic mouse model of colorectal cancer
			Design and execution
			Interpretation of results
		A xenograft model of prostate cancer metastasis
			Design and execution
			Interpretation of results
		Humanized mouse models for tumor xenografts
			Design and execution
	Ethical issues
		Turning point
	Translational significance
	World Wide Web resources
	References
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
6 The clinico-molecular approaches for detection of human papillomavirus
	Summary
	What you can expect to know
	Introduction
		Cancer
		Cervical cancer
	Historical overview
	Mistaken theories of cervical cancer causation
	The first breakthrough
		zur Hausen
	Prevalence and epidemiology of cervical cancer
		Global scenario
		Symptoms of cervical cancer
		Anatomy of female pelvis
		Types of cervical cancer
		Risk factors for cervical cancer
		Human papillomaviruses
			Genomic organization of human papillomavirus
			Transcriptional regulation of human papillomavirus
			Life cycle of human papillomavirus
			Functions of human papillomavirus oncoproteins E6 and E7
			Inactivation and degradation of p53 through the E6/E6AP complex
	Screening and diagnostic methodologies of cervical cancer
		Screening
	Methods used for screening/diagnosis of cervical cancer
		Visual methods
			Indications
		Other screening tests
		Reporting systems terminology
		Precancer classification
			WHO classification
			CIN classification (Bhambhani., 2007)
			Bethesda classification
		Cancer classification
			Colposcopy and biopsy
			New technologies
			DNA cytometry
			Human papillomavirus DNA-based screening methods (protocol)
			Urine-based noninvasive human papillomavirus DNA detection method
			Simple “paper smear” method for rapid detection of human papillomavirus infection
			Detection of HPV by multiplex PCR and RFLP
			Statistical information
			Hybrid Capture II Method
			Principle
	Applications
		Next-generation sequencing
		Clinical correlations
	Treatment
		Ablative techniques
		Excisional techniques
		Follow-up for excisional/ablative treatment
			Hysterectomy
	Stage-wise management of cervical cancer
		Treatment of microinvasive carcinoma
			Stage IA1 and IA2
		Treatment of early invasive cancer (stage IB1 and IIA %3c 4cm)
		Treatment of early bulky disease (stage IB2 and IIA %3e 4cm)
		Treatment of extensive disease: stages IIB–IIIB
		Treatment of stage IVA
		Treatment of stage IVB or recurrent disease
		Human papillomavirus vaccines
			Prophylactic human papillomavirus vaccines
			Therapeutic human papillomavirus vaccine
			Genetic-based DNA vaccine
		Issues/unanswered questions associated with human papillomavirus vaccine
			Vaccine efficacy
			Vaccine protection
			Who should be vaccinated
	Ethical issues
		With screening implementation
			Risk
			Benefit versus cost
			Patient autonomy and coercion
		With vaccine implementation
	Translational significance
		MicroRNA expression profiles in cervical cancer
		siRNAs for human papillomavirus oncogenes as potential gene therapy for cervical cancer
		Chemotherapeutic drugs and siRNAs
	Conclusion
	World Wide Web resources
		The American Cancer Society Southwest Division
		CancerNet
		People Living Through Cancer
		American Institute for Cancer Research
		American Society of Plastic and Reconstructive Surgeons
		Asian and Pacific Islander American Health Forum
		Avon’s Breast Cancer Awareness Crusade
		Cancer Information Service
		Cancer Mail
		CancerNet
		Cancer Patient Education Database
		Cancer Research Foundation of America
		Cancer source.com
	Further information
		Keywords
		Key points to remember
		Key points for effective cervical cancer screening program in low-resource settings
	Ablative techniques
	Excisional techniques
	References
	Glossary
	Abbreviations
	Long answer questions
	Answers to long answer questions
	Short answer questions
	Answers to short answer questions
	“Yes/no” type questions
	Answers to yes/no type questions
7 Human DNA tumor viruses and oncogenesis
	Summary
	What you can expect to know
	History and methods
	Transformation and oncogenesis
	History of human DNA tumor viruses and cancer
		Epstein–Barr virus
		Kaposi’s sarcoma–associated herpesvirus
			Kaposi’s sarcoma
			Primary effusion lymphoma
			Multicentric Castleman’s disease
			KSHV inflammatory cytokine syndrome
		Human papillomavirus
		Hepatitis B virus
		Human polyomaviruses
			Merkel cell polyomavirus
	Principle
		Epstein–Barr virus genome structure
		Entry into the cell
		Epstein–Barr virus lytic replication
		Epstein–Barr virus latency
		Epstein–Barr virus latent genes
			Epstein–Barr virus nuclear antigen 1
			Epstein–Barr virus nuclear antigen 2
			Epstein–Barr virus nuclear antigen 3 family
			EBNA-LP
			LMP1
			LMP2
			EBERs
			BARTs
			Micro RNAs
	Clinical correlation
		Burkitt’s lymphoma
		Nasopharyngeal cancer
		Hodgkin’s lymphoma
		Infectious mononucleosis
		X-linked lymphoproliferative disease
		Research methods and protocols
	Turning point: modeling Epstein–Barr virus infection and pathogenesis
	Current research perspectives
	Ethical issues
	Translational significance
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
8 Animal models for human disease
	Summary
	Outline
	What you expect to know
	Introduction
	Rheumatoid arthritis
		Epidemiology and etiology
		Pathogenesis
		Clinical manifestations
		Treatment
		Experimental models
			Spontaneous models
			Induced models
		Pathology of collagen-induced arthritis
		Methodology and protocols
			Clinical assessment of collagen-induced arthritis
			Histological assessment
			Radiographic evaluation
	Multiple sclerosis
		Epidemiology and etiology
		Pathogenesis
		Clinical manifestations
		Treatment
		Experimental models
			Spontaneous models
			Induced models
		Methodology and protocol
			Protocol
			Clinical evaluation
			Histology
			Immunohistochemistry
			Enzyme-linked immunosorbent assay
			Real-time polymerase chain reaction
	Ethical issues
	Translational significance
	Clinical correlations
	Conclusion
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long-answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
9 HIV and antiretroviral drugs
	Summary
	What you can expect to know
	History and methods
		Introduction
	Discovery and origin of HIV
	History of HIV and AIDS
	Global disease burden
	Clinical stages of HIV
		Classification of clinical stages
			Stage I: primary HIV infection or seroconversion stage
			Stage II: asymptomatic stage
			Stage III: persistent generalized lymphadenopathy
			Stage IV: symptomatic stage
	Molecular biology of HIV
		Envelope
		Group-specific antigen
		Long-terminal repeats
		Negative factor
		Polymerase
		Regulator of expression of viral proteins
		Transactivator of transcription
		Viral infectivity factor
		Viral protein U
		Viral protein R
	Replication: steps and drug targets
	Antiretroviral drugs
	HIV resistance and antiretroviral treatment
		Highly active antiretroviral treatment
		Salvage therapy
		Drug holiday
	New types of antiretrovirals
	Methodology and principles
		Growing HIV stock
			Principles
			Crucial steps
		Assays for antiretroviral drugs
		Monitoring antiretroviral drug toxicity
			Principles
			Special note
			Crucial steps
		Evaluating anti-HIV effects of antiretroviral drugs
			Principles
			Special note
			Crucial steps
	Understanding HIV reservoir
		Mechanism of viral latency
	NeuroAIDS: an emerging health concern
	Bone marrow transplantation: a probable cure for HIV
	Ethical issues
	Translational significance
	Clinical correlation
	Acknowledgments
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
10 Animal models in advancement of research in enteric diseases
	Summary
	What you can expect to know
	Introduction
	Animal models for enterotoxigenic Escherichia coli
		Suckling mouse model
		Infant mouse model
		Adult mouse model
		Rat model
		Adult rabbit ligated ileal loop model
	Animal models for Vibrio cholerae
		Infant mouse model
		Suckling mouse model
		Infant rabbit model
		Rabbit ligated ileal loop model
		Ileal ligated mouse model of cholera
	Animal models for Campylobacter jejuni
		Nonhuman primate model
		Ferret model
		Pig model
		Mouse model
		Rabbit model
		Chicken model
	Animal models for Shigella
		Macaque monkey model
		Pig model
		Rabbit model
		Guinea pig model
		Mouse model
	Animal models for Salmonella typhi
		Mouse model
	Animal models for nontyphoidal Salmonella
		Opium-treated guinea pig model
		Calf gastroenteritis model
		Mouse model
		Coinfection model
	Translational significance
	World Wide Web resources
	References
	Further reading
	Glossary
	Long answer questions
	Short answer questions
	Yes/no types questions
	Answers to yes/no questions
11 Chick chorioallantoic membrane assay: a 3D animal model for cancer invasion and metastasis
	Summary
	What you can expect to know
	Introduction
	History
	Principle
		Structure and function of the CAM
		Methodology
			Examples With Applications
				Tumor Growth
				Tumor Invasion
				Tumor Metastasis
				Patient-Derived Xenografts
				Response to anticancer drugs
				Advantages and limitations of the cam assay
	Conclusions
	Ethical Issues
	Translational significance
		Clinical correlations
		Turning point
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
Section II: Animal biotechnology: tools and techniques
12 Animal biotechnology as a tool to understand and fight aging
	Summary
	What you expect to know
	Introduction
		Theories of aging
			Evolutionary theories
			Molecular theories
			Cellular theories
				Cell senescence/telomere theory
			Free radical theory of aging
			System-based theories
				Neuroendocrine theory
			Caloric restriction theory
	Principle
		Reactive oxygen species—causative agent of aging
			Superoxide anion (O2−)
			Hydroxyl radical (OH)
			Hydrogen peroxide (H2O2)
			Singlet oxygen
			Peroxyl and alkoxyl radicals
			Nitric oxide and nitric dioxide
			Peroxynitrite
			Enzymatic formation
	Methodology: measurement of free radicals and methods to monitor aging
		Protein oxidation/protein carbonyl content
		Antioxidant capacity
	Common laboratory animal experimental models for aging research
		Mice
		Fish
	Polyphenols as an agent to fight aging
		Flavonoids
		Tea as antiaging compound
		Types of teas
		Tea catechins
		Health benefits of tea
		Molecular mechanisms of green tea effects
		Green tea in aging and neurodegenerative diseases
	Animal biotechnology as a tool to understand aging and fight aging
	Human aging: a translational perspective and significance
	Ethical issues related with aging research
	World wide web resources
	Clinical correlation
	References
	Further reading
	Glossary
	Abbreviations
	Long-answer questions
	Short-answer questions
	Answers to short-answer questions
	Yes/no-type questions
	Answers to yes/no-type questions
13 Multicellular tumor spheroids as in vitro models for studying tumor responses to anticancer therapies
	Summary
	What you can expect to know
	History and methods
		Introduction
	Multicellular tumor spheroids
	Historical facts toward the development of tissue culture technology from 2D and 3D cultures
		Examples where 3D culture is more beneficial over 2D culture
	Techniques for the generation of spheroids
		Hanging-drop method
		Liquid overlay method
		Microfabricated microstructures method
		Rotatory flask methods
		Surface modification-based methods
		Chip-based spheroid generation
		Emerging technologies for the generation of spheroids
	Protocol for tumor spheroid generation
	Drug treatment protocol
	Parameters to monitor drug efficacy in 3D cultures
	Radiation response of tumor cells and its modifications
	Response to anticancer drugs
	Response to photodynamic therapy
	Response to antiangiogenesis therapeutics
	Evaluation of response to immunotherapy
	Application of 3D cultures in other diseases
	Conclusions
	Ethical issues
	Translational significance
	World Wide Web resources
	References
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
14 Animal tissue culture principles and applications
	Summary
	What you can expect to know
	History and methods
		Introduction
	Development of animal cell culture
	Basic concept of cell culture
		How are cell cultures obtained?
			Organ culture
			Primary explant culture
			Cell culture
				Monolayer cultures
				Suspension cultures
		Cell passage and use of trypsin
		Quantitation
			Hemocytometer
			Electronic counting
			Other quantitation
		Reconstruction of three-dimensional structures
			Histotypic culture
			Organotypic culture
	Types of cell culture
		Primary cell culture
			Advantages and disadvantages of primary cell culture
			Anchorage-dependent/adherent cells
			Anchorage-independent/suspension cells
		Secondary cell culture
			Advantages and disadvantages of secondary cell culture
	Cell line
		Finite cell lines
		Indefinite cell lines
		Commonly used cell lines
		Advantages of continuous cell lines
	Growth cycle
		Phases of the growth cycle
			Lag phase
			Log phase
			Plateau phase
	Monitoring cell growth
		Characteristics of cell cultures
			Temperature
			pH
			Oxygen
	Cell viability
		Cytotoxicity
		Hayflick’s phenomenon
	Culture media
		Basic components in culture media
			Natural media
			Artificial media
		Serum
			Advantages of serum in cell culture medium
			Disadvantages of serum-containing medium
		Serum-free media
			Advantages of serum-free culture media
			Disadvantages of serum-free media
		Chemically defined media
		Protein-free media
	Characterization of cell lines
		Identity testing
			Karyotyping
		Purity testing
		Stability testing
		Viral testing assays
	Advantages of animal cell culture
	Disadvantages of animal cell culture
	Ethical issues
	Use of fetal bovine serum in animal culture of media
	Translational significance
	Antiviral vaccines
		Viral particles production by cell culture
		Production of virus-like particles
			Vaccines based on virus-like particles
		Human papilloma virus vaccine
	Recombinant therapeutic proteins
		Main therapeutic proteins
			Cytokines
		Applications of interferons
			Growth factors
			Hormones
			Therapeutic enzymes
			Blood coagulation factors
			Antibodies
	Gene therapy
		Importance of cell culture in gene therapy
		Clinical correlation
	Biopesticides
		Baculovirus production in animal cell culture
		Cell lines for biopesticide production
		Viral mutant formation in cell culture
	Monoclonal antibodies
	Stem cells
		Culturing embryonic stem cells in the laboratory
			Microfluidics three-dimensional culture
			Applications
			Organ-on-a-chip technology
			Tissue models on a chip
			Liver-on-a-chip
			Tumor-on-a-chip
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
15 Concepts of tissue engineering
	Summary
	What can you expect to know?
	Introduction
	History
	Basic approach to tissue engineering: principles and methodology
		Cells
		Scaffolds
		Media
		Bioreactors
		Methodology
	Scaffold design
	Materials for scaffolds
	Scaffold fabrication methods
		Fiber bonding
		Solvent casting and particulate leaching
		Melt molding
		Membrane lamination
		Phase separation
		Gas foaming
		Polymer ceramic composite foam
		Solid-free form techniques
			Selective laser sintering
			Stereolithography
			Fused deposition modeling
			3D Printing
		Pressure-assisted micro-syringe method
		Freeze-drying
	Examples of tissue-engineered organs
		Skin
		Pancreas
		Liver
		Kidney
		Bone/cartilage
		Nerves
		Blood vessels
	Tissue engineering using stem cells
	Issues and challenges
	Ethical issues
	Translational significance
	World wide web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long-answer questions
	Short-answer questions
	Answers to short-answer questions
	Kindly state yes or no against the following statements
	Answers to yes/no statements
16 Nanotechnology and its applications to animal biotechnology
	Summary
	What you can expect to know
	History and methods
		Introduction
	Methodologies
		Nanotools and nanotechniques
			SPM techniques
			Raman spectroscopy and imaging
			AFM–Raman confocal hybrid systems
		Chemical modification of AFM probes
		Nanostructural features of animal cells and tissues
		Nanomechanical properties of animal cells and tissues
		Nanomanipulation
		Nanofabrication
	Examples of nanotechnology applications to animal biotechnology
		Nanoparticles in animal production
		Nanosurgery
			AFM as a diagnostic tool to identify orthopoxvirus in animals
			Frictional response of bovine articular cartilage
			Microstructure and nanomechanical properties of cortical bone osteons from baboons
			Use of calf thymus DNA for cancer experiments
			Characterization of mitochondria isolated from normal and ischemic hearts in rats
			Polymorphism and ultrastructural organization of prion protein
			Ultrastructural investigation of animal spermatozoa using AFM
			Multifactor analysis of living animal cells for biotechnology and medicine
	Ethical issues
	Translational significance
		Clinical correlations
	Acknowledgments
	World wide web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long Answer Questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to “Yes/no” questions
17 Antibodies: monoclonal and polyclonal
	Summary
	What you can expect to know
	History and methods
		Introduction
	Tiselius and Kabat’s experiment
	History
	Elucidation of immunoglobulin structure
		Edelman’s experiment
		Porter’s experiment
		Nisonoff’s experiment
		Conclusion from papain and pepsin digestion
		Turning point
	Immunoglobulin G: a prototype for immunoglobulin
	Polyclonal antibody versus monoclonal antibody
		Polyclonal antibodies
		Monoclonal antibodies
		Naming monoclonal antibodies
			Prefix
			Infix-1
			Infix-2
			Additional words
	Antibodies as therapeutics: adverse effects
		Serum sickness
		Human antimonoclonal antibody response
		Human antichimeric antibody response
		Human antihumanized antibody response
	Applications of antibodies
		Therapeutic applications
		Analytical applications
		Preparative applications
	Methodology, principles, and protocols
		Polyclonal antibodies
			Principle
			Methodology and rationale for PoAb production
				Step 1: antigen preparation
				Step 2: immunization of animals
				Step 3: antibody titer
				Step 4: purification and identification
		Monoclonal antibodies
			Principle
			Methodology and rationale of MoAb production
				Step 1: immunization of mouse
				Step 2: preparation of splenocytes
				Step 3: fusion of cells
				Step 4: selection of hybrid cells
				Step 5: selection of clones
				Step 6: expansion of clone
				Step 7: purification
		Antibody titration
	Biochemical pathway: hybridoma selection
	Ethical issues
	Camelid nanobodies/single-domain antibodies/variable domain of camelid heavy chain only antibody
	Translational significance
	Clinical correlations
	World Wide Web resources
	Acknowledgments
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type question
	Answers to yes/no type questions
18 Molecular markers: tool for genetic analysis
	Summary
	What can you expect to know
	Introduction
	Methodology
		Restriction fragment length polymorphism
			Steps involved in restriction fragment length polymorphism analysis
			Applications of restriction fragment length polymorphism
		Allele-specific oligonucleotide
			Applications of allele-specific oligonucleotide
		Allele-specific polymerase chain reaction
			Applications of allele-specific polymerase chain reaction
		Single-strand conformation polymorphism
			Applications of single-strand conformation polymorphism
		Sequence-tagged site
			Applications of sequence-tagged site
		Random amplified polymorphic DNA
			Steps involved in random amplified polymorphic DNA analysis
			Applications of random amplified polymeric DNA analysis
		Restriction landmark genome scanning
			Applications of landmark genome scanning
		Single nucleotide polymorphisms
			Applications of single-nucleotide polymorphisms
		Amplified fragment length polymorphism
			Steps involved in amplified fragment length polymorphism analysis
			Applications of amplified fragment length polymorphism
		Methylation-sensitive amplification polymorphism
			Applications of methylation-sensitive amplification polymorphism
		Miniature inverted-repeat transposable element
			Applications of miniature inverted-repeat transposable element
		Microsatellites
			Simple sequence repeats/Short tandem repeats/Simple sequence tandem repeats
			Applications of simple sequence repeats
			Inter simple sequence repeats
			Applications of inter simple sequence repeats
			Variable number of tandem repeat markers
			Applications of variable number of tandem repeat markers
			Sequence tagged microsatellite site
			Applications of sequence-tagged microsatellite site
			Simple sequence length polymorphisms
			Applications of sequence length polymorphisms
			Example of microsatellites
			Advantage of microsatellite marker
			Disadvantages of microsatellite marker
			Some general applications of microsatellites
				Parentage determination
				Determination of twin zygosity and freemartinism
				Identification of disease carrier
	Ethical issues
	Translational significance
	Clinical correlations
	Turning point
	World wide web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long-answer questions
	Short-answer questions
	Answers to short questions
	Yes/no-type questions
	Answers to yes/No-type questions
19 Ribotyping: a tool for molecular taxonomy
	Summary
	What you can expect to know
	History and methods
		Introduction
	Historical developments in bacterial taxonomy
	Typing methods used for bacterial systematics
		Phenotypic typing methods
		Genotypic typing methods
	Basis of using rRNA and rRNA genes as taxonomic tools
		Organization of the ribosomal operon
	Different techniques of ribotyping
		Conventional ribotyping
		Selection of restriction endonuclease for ribotyping by sequence analysis (in silico)
		Automated ribotyping
		Polymerase chain reaction ribotyping
			Polymerase chain reaction ribotyping and endonuclease subtyping
			Polymerase chain reaction ribotyping followed by sequencing of ISR
			Amplified ribosomal DNA restriction analysis
			Terminal restriction fragment length polymorphism of 16S rRNA gene
			Long polymerase chain reaction ribotyping
			Broad-range polymerase chain reaction ribotyping
			Limitations of polymerase chain reaction ribotyping
			Ribosomal DNA sequence analysis
				In situ hybridization targeted to detect rRNA
			Clone-fluorescent in situ hybridization
			Catalyzed reported deposition fluorescent in situ hybridization
			Recognition of individual gene fluorescent in situ hybridization
			Stable isotope-labeled probing of rRNA and rDNA
			Peptide nucleic acid probes
	Limitations of ribotyping
		Other genotyping methods
			Multilocus sequence typing
			Whole genome sequencing
			Single cell sequencing
			Metagenomics
	Future perspectives
	Ethical issues
	Translational significance
		Clinical significance
	World Wide Web resources
	References
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
20 Next generation sequencing and its applications
	Summary
	What you can expect to know
	Introduction
	History of DNA sequencing
	Generation of sequencing technologies
	Principle of Sanger sequencing versus NGS
	NGS technologies: Initial phase
		Pyrosequencing technology
		Sequencing by ligation technology
		True single molecule sequencing
	NGS technologies: recent phase
		Reversible terminator technology
		Ion semiconductor sequencing
		Single molecule real time sequencing
	Other newer sequencing technologies
		Nanopore sequencing
		Polony-based sequencing technology
		DNA nanoball sequencing
	Downstream bioinformatics
		Primary analysis
		Secondary analysis
		Tertiary analysis
	General principles of NGS methods in various applications
		Whole genome de novo sequencing
		Whole genome resequencing
		Targeted resequencing
		Transcriptome sequencing
		Amplicon sequencing
		Chromatin immunoprecipitation DNA sequencing
		Small RNA sequencing
	Animal biotechnology and the cattle genome
	Applications of NGS in animal biotechnology
		Evolutionary research
		Epigenetics
		Metagenome sequencing
		Ancient DNA
		Genomic variability, SNP/copy number variations discovery
		Beef cattle selection
		Animal breeding and improvement of livestock productivity and health
		Food, safety and nutrition
		Transgenics
	Applications of NGS in human health
		Cancer research
		Genetic disorders
		Human microbiome
		Pre and postnatal diagnoses
		Infectious diseases
		Personalized medicine
	Clinical correlation
	Translational significance
	Ethical issues
	Future perspectives
	Challenges
		Web resources for NGS
	World wide web resources
	References
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/No type questions
	Answers to Yes/No type questions
21 Biomolecular display technology: a new tool for drug discovery
	Summary
	What you can expect to know
	Introduction
	Principle
	Necessity: small molecule versus biomolecular (biologics) drugs
	Methodology: biomolecular display technologies
		Phage display
			Selection of peptides
		Ribosome display
			Selection of peptides
		mRNA display and “head-to-head” mRNA display
			Selection of peptides
		Other display systems
		A general method for discovery of functional peptide aptamers
	Translational significance
	World Wide Web resources
	Conclusion and future perspective
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
22 In silico disease model: from simple networks to complex diseases
	Summary
	What you can expect to know
	Bioinformatics in animal biotechnology
	Bioinformatics and systems biology
	Common computational methods in systems biology
	Experimental methods in systems biology
	Protein–protein interactions
	Transcriptional control networks
	Signal transduction networks
	Mathematical modeling techniques
	Concept of modeling
	In silico models of cells
		In silico metabolic modeling
		In silico modeling of disease: in practice
		In silico models of cancer
		In silico models and inflammatory response syndrome in trauma and infection
	Applications of in silico disease modeling
		Infectious diseases
			Triad of infectious diseases as the source of parameters for in silico modeling of infectious diseases
			Parameters for in silico modeling of infectious diseases
				Parameters derived from characteristics of agent
				Parameters derived from characteristics of host
				Parameters derived from characteristics of environment
			Infectious diseases in silico model proper, a typical approach/scenario
			Specific examples of infectious diseases in silico model
		Model of bacterial and viral dynamics
			Challenges in in silico modeling of infectious diseases
		Neuronal diseases
			Pathophysiology of neuronal diseases as the source of parameters for in silico modeling of neuronal diseases
			Parameters for in silico modeling of neuronal diseases
				Parameters derived from characteristics of nervous cell
				Parameters derived from characteristics of signaling chemicals and body electrolytes
				Parameters derived from host/organism factors
				Parameters derived from environmental factors
			Neuronal disease in silico model proper, a typical approach/scenario
			Specific examples of neuronal disease in silico model
			Possible limitations of in silico modeling of neuronal diseases
	Conclusion
	References
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
Section III: Animal biotechnology: applications and concerns
23 Transgenic animals in research and industry
	Summary
	What you expect to know
	Introduction
	Creating transgenic animals
		Construction of a transgene
		Microinjection
		Embryonic stem cell transfer
		Retrovirus-mediated gene transfer
		Gene knockdown and RNA interference
		CRISPR/Cas9 systems
		Screening for transgenic positives
			Transgenesis versus cloning
	Transgenic animals as disease models
		Oncomouse
		AIDS mouse
		Alzheimer’s mouse
		Parkinson’s fly
	Transgenic animals as biological models
		ANDi (monkey)
		Doogie (the smart mouse)
		Supermouse
		Youth mouse
		Influenza-resistant mouse
	Transgenic animals as xenotransplanters
	Transgenic animals as food source
		Superpig
		Superfish
	Transgenic animals for drug and industrial production
	Transgenic animals’ impact on the environment
	Patenting transgenic animals
	Ethics in transgenesis
	FDA guidelines on genetically engineered animals
	Translational significance
	Clinical correlations
	Turning point
	World Wide Web resources
	References
	Further reading
	Glossary
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type of questions
	Answers to yes/no type questions
24 Role of cytogenetics and molecular genetics in human health and medicine
	Summary
	What you can expect to know
	Introduction
	Cytogenetics: an overview
	Chromosome morphology and classification
		Nomenclature
	Chromosomal disorders
		Structural abnormalities
		Chromosome breakage and fragile sites
	Methodology: application of different cytogenetic techniques in the diagnosis of genetic disorders
		Identification of chromosomes and karyotyping
		Fluorescence in situ hybridization
		Array-comparative genomic hybridization
		Principle
	Molecular genetics: an overview
		Hereditary material
			The structure of DNA
			Single nucleotide polymorphism
		Single-gene disorders
		Multigenic and multifactorial disorders
		Mitochondrial disorder
			Epigenetic disorders
	Methodology: application of different molecular techniques for diagnosis of genetic disorders
		Southern blotting
			Real-time polymerase chain reaction
			Quantitative fluorescent polymerase chain reaction
		DNA sequencing
		Principle
		Case study 1
			Prenatal diagnosis
	Ethical issues
	Translational significance
		Promising future
	World Wide Web resources
	References
	Further Reading
	Glossary
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
25 Antibodies and their applications
	Summary
	What you can expect to know
	History and methods
		Immunodiagnostics: role of antibodies
			Introduction
		History
		Antigens and antibodies
		Polyclonal and monoclonal antibodies
			Principle
		Hybridoma technology and methodology
		Application of monoclonal antibodies
			Immunosorbent chromatography
			Blood typing reagent
			More sensitive in RIA
			Positive and negative selection of cells
			Negative selection
			Positive selection
			Diagnostic and therapeutic application
			Choice of linkers
			Chimeric and humanized antibodies
				Methodology
			Phage display technology: screening recombinant antibody libraries
				Methodology
		Antibody constructs
		Diabody
		Human antibodies from transgenic mouse
			Methodology
		Bispecific antibodies
	Ethical issues
		Clinical corelation of the chapter content
	Translational significance
	World Wide Web resources
		Pros
		Cons
		Websites
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/No type questions
	Answers to Yes/No type questions
26 Vaccines: present status and applications
	Summary
	What you can expect to know
	Introduction
	Types of vaccines
		Traditional vaccines
			Live, attenuated vaccines
				Methodology
				Advantages of live, attenuated vaccines
				Disadvantages of live, attenuated vaccines
			Inactivated whole virus vaccines
				Methodology
				Advantages of inactivated whole virus vaccines
				Disadvantages of inactivated whole virus vaccines
			Toxoid vaccines
				Methodology
			Subunit vaccines
				Methodology
			Conjugate vaccines
			DNA vaccines
				Methodology
					Construction of DNA vaccine
				Action mechanisms of DNA vaccines
				Advantages of DNA vaccines
				Limitations of DNA vaccines
				Future of DNA vaccines
			Recombinant vector vaccines
	Molecular farming using plants as bioreactor
	Advancement in vaccine adjuvants
		Alum salts
		MF59 and AS03
		Virosomes
		Toll-like receptors agonist
	Immunostimulating complexes
	Future challenges in vaccine development
		Foremost infectious disease problems
		Infectious disease threats
	Ethical issues
	Mandates
	Vaccine research and testing
	Informed consent
	Access issues
	Translational significance
	World Wide Web resources
	Protocols
		Protocol for the development of vaccines
	References
	Further reading
	Glossary
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
27 Perspectives on the human genome
	Summary
	What you can expect to know
	History and methods
		Introduction
	Human genome sequencing project
		History
		Human genome: organization and perspective
		Complexity of human genome
		Gene content
			Development of next-generation sequencing technology
			Principle and history of human genome sequencing
			Methodology used for initial Human Genome Project
			Examples with applications
			The 1000 Genomes Project
			Genome-wide association studies
	Ethical issues
	Translational significance
		Precision medicine
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
28 Marine resources and animals in modern biotechnology
	Summary
	What can you expect to know
	Introduction
	Marine biological diversity
		Historical background
		Biotechnologically important marine animals
	Advances in mariculture
		Captive rearing technology
		Feed technology for culture fisheries
		Marine food and agri-biotech industry
		Marine ornamental fish trade
		Chromosomal manipulation in marine fish
			Polyploidy
			Gynogenesis
			Androgenesis
			Cryopreservation of gametes
	Genetic engineering technology
		Marine genomics
		Marine transcriptomics
		Marine proteomics
	Marine metagenomics
	Marine animals and nanotechnology
		Marine animals in synthesis of nanomaterials
		Application of nanotechnology in aquaculture and fisheries
	Pharmaceuticals and therapeutics from marine animals
		Marine natural products of animal origin
		Commercial bioproducts from marine organisms
		Green fluorescent protein from jelly fish and its application
		Red fluorescent protein from corals and its application
	Ethical issues
	Translational significance
	Future directions
	World wide web resources
	References
	Further Reading
	Glossary
	Long-answer questions
	Short-answer questions
	Answers to short answer questions
	Yes/no-type questions
	Answers to yes/no-type questions
29 Nanotechnology and detection of microbial pathogens
	Summary
	What you can expect to know
	Introduction
	Indicators of microbial water quality
	Need for detection of waterborne and foodborne pathogens
	Conventional methods to detect fecal indicator organism and other pathogenic bacteria
		Most probable number method
		Membrane filtration method
		Defined substrate methods
		Rapid detection using chromogenic substrates
		Immunological methods
			Antibodies-based detection
			Immunomagnetic separation and other rapid culture-based methods
	Molecular methods based on genetic signature of target pathogen
		Polymerase chain reaction technique and quantitative polymerase chain reaction
			Nucleic acid microarrays
			Next-generation sequencing
			Digital polymerase chain reaction
			Loop-mediated isothermal amplification
	Nanotechnology and promises
		Metallic nanoparticles
			Gold nanoparticles–based colorimetric assays
			Silver nanoparticles colorimetric assays
			Quantum dots
			Silica/other nanomaterials
	History
	Detection principle
	Methodology
		Synthesis of gold nanoparticles
		Computation of single-stranded DNA sequences for functionalization of gold nanoparticles
		Functionalization of gold nanoparticles with thiol-modified DNA
	Examples of application of gold nanoparticles and few examples of silver nanoparticle and quantum dots for detection of bac...
		Colorimetric detection of DNA of shiga toxin producing E. coli using bioconjugated gold nanoparticles
		Colorimetric detection of enterotoxigenic E. coli gene using gold nanoparticle probes
		Clinical significance of nanoparticle-based detection
	Ethical issues
	Translational significance
	Futuristic approach
	World Wide Web resources
	Clinical correlations
	Turning point
	Awards/recognitions
	Acknowledgment
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
30 Herbal medicine and biotechnology for the benefit of human health
	Summary
	What you can expect to know
	Introduction
		Traditional medicine
		Ancient system of medicine
	Methodology
		Investigation of medicinal plants
			Extraction
			Chemical screening
			Biological assays
	Identification, quantification, and characterization of bioactive compounds
	Biotechnological approaches for herbal drug production
		Organ cultures
		Callus cultures
		Suspension cultures
		Case study: Lantana camara L
	Opportunities and challenges
	Conclusions and outlook
	Ethical issues
	Translational significance
		Clinical correlation
		Turning point
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
31 Enzyme inhibition assay for metabolic disorders—exploring leads from medicinal plants
	Summary
	What you can expect to know
	Introduction
		Lead molecules from herbs
		Principle
	Methodology
		Maceration
		Infusion
		Digestion
		Decoction
		Percolation
		Hot continuous extraction
		Aqueous alcoholic extraction by fermentation
		Countercurrent extraction
		Ultrasound-assisted extraction (sonication)
		Microwave-assisted extraction
		Supercritical fluid extraction
		Bioguided fractionation of extract and isolation of phytoconstituents
	Examples with applications
		Instrumental techniques for quality evaluation
	In vitro enzyme inhibition assays for screening of medicinal plants in metabolic disorders
		Metabolic disorders
		Enzyme inhibition: a target of drug discovery
		Enzymes involved in metabolic disorder: its significance in inhibition
			Pancreatic lipase
			3-Hydroxy-3-methylglutaryl coenzyme A reductase
			Glucosidase
			Aldose reductase
			Carbonic anhydrase
	Medicinal plants used in metabolic disorders
	Ethical issues
	Translational significance
	World Wide Web resources
	Acknowledgment
	References
	Glossary
	Abbreviations
	Long answer questions
	Answers to long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
32 Safety assessment of food derived from genetically modified crops
	Summary
	What you can expect to know
	History and methods
		Introduction
	Rationale for the allergenicity assessment of genetically modified Foods
	Mechanism of food protein-induced allergenicity
	Simulated gastric fluid assay
		How simulated gastric fluid assay works
		Components of simulated gastric fluid
		General protocol of the simulated gastric fluid assay
		Factors relevant to gastrointestinal digestion of allergens
		Supportive and negative evidence of simulated gastric fluid
		General protocol of simulated intestinal fluid assay
		Effect of assay conditions on protein stability in simulated intestinal fluid assay
		Supportive and negative evidence of simulated intestinal fluid assay
		Contradictory result of simulated intestinal fluid digestibility of food proteins
	Thermal treatment assay
		Mechanism of thermal treatment assay
		Standard protocol for thermal treatment assay
		Functional stability of proteins and importance of thermal stability assay
		Contradictory result in thermal treatment procedure
	Ethical issues
		Clinical correlation
	Translational significance
	Acknowledgment
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/no type questions
	Answers to yes/no type questions
33 Correlating Ayurveda and biotechnology: approaches for the 21st century and beyond
	Summary
	What you can expect to know
	Introduction
		Principles of Ayurveda
			Ayurvedic physiology (Sharir kriya)
		Panchamahabhuta
		Doshas
			Manasik doshas
			Types of vata (Vagbhatta, 2001b)
			Types of pitta
			Types of kapha
		Dhatus and Mala (tissues and waste)
			Characteristics of Dhatus
		Agni (energy)
		Prakriti (psychosomatic constitution)
			Traits of Vata Prakriti person
			Traits of Pitta Prakriti person
			Traits of Kapha Prakriti person
		Aahar (diet)
			Inappropriate food
			Diet of a Vataj Prakriti person
			Diet of a Pittaj Prakriti person
			Diet of a Kaphaj Prakriti person
		Vihar (behavioral suggestions)
			Dinacharya
			Ritucharya
			Hemanta ritucharya (mid-November to mid-January)
			Shishir ritucharya (mid-January to mid-March)
			Vasanta ritucharya (mid-March to mid-May)
			Grishma ritucharya (mid-May to mid-July)
			Varsa ritucharya (mid-July to mid-Sep)
			Sharad ritucharya (mid-Sep to mid-Nov)
	Ageing process
	Prakriti and genomics
	World Wide Web resources
	Way forward
	Acknowledgment
	References
	Further reading
	Glossary
	Long answer questions
	Short answer questions
	Yes/no type questions
	Answers to yes/no questions
34 Nanoparticle synthesis harnessing benign green routes
	Summary
	What you can expect to know
	Introduction
	History
	Principle
	Methodology
		Materials required
		Methods
		Nanoparticle production using algae
		Nanoparticle production using plants
		Nanoparticle production using biomolecules
	Examples
		Bacteria-mediated synthesis of nanoparticles
		Algae-mediated synthesis of nanoparticles
		Fungi-mediated synthesis of nanoparticles
		Plant-mediated synthesis of nanoparticles
		Biomolecule-mediated synthesis of nanoparticles
		Alginate-mediated synthesis
		Pectin-mediated synthesis
		Microbial exopolysaccharide-mediated synthesis
		Nanoparticle synthesis based on tree gums
		Gum ghatti
		Gum arabic (gum acacia)
		Neem gum
		Microbial biosurfactant–mediated synthesis of nanoparticles
		Enzyme-mediated synthesis of nanoparticles
		Pigment-mediated synthesis of nanoparticles
		Honey-mediated synthesis of nanoparticles
	Protocol
		Nanoparticle production using bacteria
		Synthesis using fungal culture supernatant
		Synthesis using fungal biomass
		Nanoparticle production using biomolecules
	Ethical issues
	Translational significance
	World Wide Web resources
	Turning point
	Acknowledgement
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/No type questions
	Answers to Yes/No type questions
35 Ethical issues in animal biotechnology
	Summary
	What you can expect to know
	History and methods
		Introduction
	A brief overview of ethical thoughts and principles
		Virtue ethics
		Deontological (duty-based) ethics
		Consequentialist ethics
	Principles
	Methodology
	Application of ethics in animal biotechnology
	Ethical concerns in animal biotechnology
		Intrinsic concerns
			Religious intrinsic critique of biotechnology
			Religious critique of human stem cell research
			Religious critique of human reproductive cloning
			Religious critique of in vitro fertilization
			Secular intrinsic objections to biotechnology
			Public perceptions about genetic modifications in biotechnology
		Extrinsic concerns
			Animal welfare
			Human health
				Nature of risk
				Public health ethics and animal biotechnology
			Environmental concerns
				Ethical positions
				Concerns posed by transgenic technology
			Precautionary principle
	Some challenging ethical issues in animal biotechnology
		Chimeras
		Animal biopharming
			Risks to human health
			Food chain contamination
			Escape of biopharm animals
			Horizontal gene transfer
			Welfare issues of biopharm animals
				Genome editing
		Constitution of ethics committees
	Translational significance
		Human therapeutic cloning and other techniques in animal biotechnology
	Conclusions
	World Wide Web resources
	References
	Glossary
	Abbreviations
	Long-answer questions
	Short-answer questions
	Answers to short-answer questions
	Yes/no-type questions
	Answers to yes/no-type questions
36 Approaches to the humane euthanasia of research animals
	Summary
	What you can expect to learn
	Introduction and background
	Proper training of personnel
	Methodology, equipment, and principles
	Chemical methods
		Injectable agents
		Inhalant agents
	Physical methods
		Unacceptable methods
	Special considerations: fetuses and neonate animals
	Ethical issues
	Translational significance
	World Wide Web resources
	References
	Further reading
	Glossary
	Abbreviations
	Long answer questions
	Short answer questions
	Answers to short answer questions
	Yes/No type questions
	Answers to Yes/No type questions
Index
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