Manual of Techniques in Invertebrate Pathology

Dedication

Contributors

Preface to first edition

Preface to second edition

Chapter I. Initial handling and diagnosis of diseased invertebrates

1 Introduction

2 Collection of diseased invertebrates and entomopathogens

3 Examination in the laboratory

4 Safety considerations

Acknowledgments

REFERENCES

Chapter II. Basic techniques in insect virology

1 Introduction

2 Brief overview of the different groups of insect viruses

3 Identification

4 Virus propagation

5 Virus quantification

6 Quantification of virus activity (virulence)

7 Cell culture techniques

8 Reverse genetics

REFERENCES

Glossary

Appendix: Recipes for stains

Chapter III. Isolation, culture, preservation, and identification of entomopathogenic bacteria of the Bacilli

1 Introduction

2 Isolation

3 Cultivation

4 Preservation

5 Identification

6 Concluding remarks

Acknowledgments

Disclaimer

REFERENCES

Appendix

Chapter IV. Bioassay of bacterial entomopathogens against insect larvae

1 Introduction

2 Laboratory bioassay of Bacillus pathogens against lepidopteran larvae

3 Bioassay of Bacillus thuringiensis against dipteran larvae

4 Bioassay of Bacillus thuringiensis against other orders

5 Bioassay of other entomopathogens

6 Conclusions

Acknowledgments

REFERENCES

Appendix: artificial diet recipes

Chapter V. Bacteria for use against soil-inhabiting insects

1 Introduction

2 Isolation

3 Identification

4 Propagation and production

5 Bioassay

6 Preservation

REFERENCES

Appendix 1

Chapter VI. Identification of entomopathogenic fungi

1 Introduction

2 Quick guide to light microscopy

3 Key to major genera of fungal entomopathogens

4 Major genera of fungal entomopathogens

REFERENCES

Notes

Brief glossary of mycological terms

Appendix: recipes of stains and reagents

Chapter VII. Laboratory techniques used for entomopathogenic fungi: Hypocreales

1 Introduction

2 Detection and characterization

3 Inoculum propagation and preparation

4 Quantification

5 Infection and bioassay

6 Assessment of environmental parameters

Acknowledgments

REFERENCES

Appendix

Chapter VIII. Mass production of entomopathogenic Hypocreales

1 Introduction

2 Culture maintenance

3 Solid Substrate Production of Aerial Conidia

4 Liquid Fermentation Production

Acknowledgments

REFERENCES

Appendix

Chapter IX. Methods for study of the Entomophthorales

1 Introduction

2 Collecting and preparing specimens

3 Species identification

4 Isolation

5 In vitro culture and production

6 In vivo culture and production

7 Bioassays

Acknowledgments

REFERENCES

Appendix: media

Chapter X. Preservation of entomopathogenic fungal cultures

1 Introduction

2 Preservation at temperatures above freezing

3 Preservation by freeze-drying (lyophilization)

4 Preservation in a frozen state

5 Record keeping for storage facilities

6 Other less common preservation techniques

REFERENCES

Chapter XI. Research methods for entomopathogenic microsporidia and other protists

1 Introduction

2 Identification

3 Techniques for managing spores

4 Molecular and proteomic techniques

5 Propagation and production

6 Controlling infections in insect colonies

7 Bioassays and experimental infections

Acknowledgments

REFERENCES

Chapter XII. Nematode parasites, pathogens and associates of insects and invertebrates of economic importance

1 Introduction

2 Major groups of nematodes associated with insects and other invertebrates of economic importance

3 Sampling techniques

4 Diagnostic methods

5 Staining methods

6 Culturing methods

7 Nematode quantification methods

8 Preservation methods

9 Shipping and handling of samples

10 Bioassays with entomopathogenic nematodes and symbiotic bacteria

Acknowledgments

REFERENCES

Appendix I

Appendix II

Chapter XIII. Pathogens and parasites of terrestrial molluscs

1 Introduction

2 General techniques for working with terrestrial mollucs

3 Major groups of pathogens and parasites associated with terrestrial molluscs

Acknowledgments

REFERENCES

Appendix

Chapter XIV. Testing the pathogenicity and infectivity of entomopathogens to mammals

1 Introduction

2 Definition of infection

3 Definition of persistence

4 Mammalian safety tests and philosophy

5 Animal procurement and housing

6 Modified tier scheme

7 Conclusion

Acknowledgments

REFERENCES

Chapter XV. Complementary techniques: preparations of entomopathogens and diseased specimens for more detailed study using microscopy

1 Introduction

2 Light microscopy

3 Electron microscopy

B Scanning electron microscopy (SEM)

Disclaimer

Acknowledgments

REFERENCES

Appendix

Index

Color Plates