Entomopathogenic Nematodes and Their Symbiotic Bacteria: A Laboratory Manual (Springer Protocols Handbooks)

Foreword
Preface
Acknowledgments
Contents
About the Authors
Chapter 1: Safety Guidelines
	1 General Instruction of Laboratory Practices
	2 Safety Measures While
		2.1 Handling Instruments
		2.2 Handling Chemicals
		2.3 Handling Glassware
	3 Personal Protection Safety Rules
	4 Lab Infrastructural Facilities
Chapter 2: Introduction
Chapter 3: Historical Aspects
	1 Evolution
	2 Biogeography
		2.1 Geographical Distribution
		2.2 Habitat Distribution
	3 Systematic Position
		3.1 Steinernema
		3.2 Heterorhabditis
		3.3 Xenorhabdus
		3.4 Photorhabdus
Chapter 4: Nematode-Bacterium Symbiosis
	1 Steinernema and Their Xenorhabdus Symbiont
	2 Heterorhabditis and Their Photorhabdus Symbiont
Chapter 5: Biology
	1 Life Cycle
	2 Bacterial Metabolites
	3 Biocontrol Potential
Chapter 6: Nematode Behavior
	1 Host-Seeking Behavior
	2 Foraging Strategies
		2.1 Ambush Foraging Strategy
			2.1.1 Standing Behavior
			2.1.2 Jumping Behavior
		2.2 Cruise Foraging Strategy
			2.2.1 Crawling Behavior
		2.3 Intermediate Foraging Strategy
	3 Foraging Assays
		3.1 Standing and Jumping Behavior
		3.2 Attachment Rate [134]
		3.3 Effect of Substrate [134]
		3.4 Host Finding [134]
Chapter 7: Chemical Ecology
	1 Chemical Cues
	2 Tritrophic Interactions [148]
		2.1 Materials Required
		2.2 Plants, Insects, and Nematodes
			2.2.1 Procedure
		2.3 Collection and Assaying of EPN Volatiles
		2.4 Larval Performance Analysis
		2.5 Insect Herbivore Oviposition Preference Analysis
	3 Volatile Cues
		3.1 Detection of Volatiles Emitted by Insect Larval Hosts
			3.1.1 Materials Required
			3.1.2 Procedure
	4 Olfactometer Assay [139]
		4.1 Materials Required
		4.2 Procedure
	5 Chemotaxis Assay
		5.1 Chemotaxis for EPN Behavior [146, 151, 152]
			5.1.1 Materials Required
			5.1.2 Procedure
		5.2 Chemotaxis to Host Volatiles [141, 154, 155]
			5.2.1 Materials Required
			5.2.2 Procedure
		5.3 Jumping Assay [156]
			5.3.1 Materials Required
			5.3.2 Procedure
		5.4 Host Chemotaxis Assay [156]
			5.4.1 Materials Required
			5.4.2 Procedure
		5.5 Nematode Chemotaxis Using Microfluidics [157]
			5.5.1 Materials Required
			5.5.2 Procedure
			5.5.3 Procedure
Chapter 8: Laboratory Techniques for Entomopathogenic Nematodes
	1 Host Insect Rearing
		1.1 Rearing of Galleria mellonella (Greater Wax Moth)
			1.1.1 Life Cycle
				Egg
				Larva
				Pupa
				Adult
			1.1.2 Rearing
				Materials Required
				Preparation of Diet
				Procedure
		1.2 Rearing of Tenebrio molitor (Mealworm)
			1.2.1 Life Cycle
				Egg
				Larva
				Pupa
				Adult
			1.2.2 Rearing
				Materials Required
				Preparation of Diet
				Procedure
		1.3 Rearing of Spodoptera litura (Tobacco Cutworm or Cotton Leaf worm)
			1.3.1 Life Cycle
				Egg
				Larva
				Pupae
				Adult
			1.3.2 Rearing
				Materials Required
				Preparation of Diet
			1.3.3 Procedure
	2 Collection and Processing of Soil Samples
		2.1 Soil Samples
		2.2 Soil Sampling Approach
			2.2.1 Random Sampling
			2.2.2 Stratified Sampling
				Materials Required
				Procedure
	3 Isolation and Storage of Soil Nematodes
		3.1 Isolation Techniques of Soil Nematodes Depend On
		3.2 Direct Method
			3.2.1 Baermann Funnel Technique [158] (Fig. 7)
				Materials Required
				Procedure
			3.2.2 Flotation Technique [159]
				Materials Required
				Procedure
		3.3 Indirect Method
			3.3.1 Insect-Baiting Technique [160]
				Materials Required
				Procedure
			3.3.2 Modified Baiting Technique [161]
				Materials Required
				Procedure
		3.4 Collection of Nematode Infective Juveniles from Infected Cadavers
			3.4.1 White Trap Method [162]
				Materials Required
				Procedure
			3.4.2 Modified White Trap Method [163]
				Materials Required
				Procedure
				Harvesting
		3.5 Koch´s Postulates [163]
			3.5.1 Isolation of Parasites from Infected G. mellonella
			3.5.2 Inoculation of IJs to a Susceptible Host
		3.6 Counting of Nematodes
			3.6.1 Quantification Methods
				Direct Count Method [164]
					Materials Required
					Procedure
				Dilution Method [164]
					Materials Required
					Procedure
		3.7 Storage of Soil Nematodes
			3.7.1 Materials Required
			3.7.2 Procedure
	4 Morphological Characterization
		4.1 Light Microscopy Studies
			4.1.1 Handling Nematodes
				Materials Required
				Procedure
			4.1.2 Preparation of Nematode Mounts
				Step 1: Killing and Fixing of Nematodes
					Materials Required
					Procedure
				Another Method of Killing Nematodes
					Materials Required
					Procedure
				Fixatives
					Materials Required
				Procedure
				Step 2: Processing Nematodes to Glycerin [166]
					Materials Required
					Procedure
				Step 3: Mounting Nematodes
					Materials Required
					Procedure
					Nematode Stages
			4.1.3 Diagnostic Characters [62]
				Family Steinernematidae
					Diagnosis
					Female
					Male
					Infective Juvenile
					Identification of Steinernema Species
					Key to Species of the Genus Steinernema (Fig. 12)
				Family Heterorhabditidae
					Diagnosis
					Hermaphroditic Female
					Amphimictic Female
					Male
					Infective Juvenile
					Identification of Heterorhabditis Species
					Key to Species of the Genus Heterorhabditis (Fig.