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Manual of Techniques in Invertebrate Pathology - 2nd Edition - ISBN: 9780123868992, 9780123869005

Manual of Techniques in Invertebrate Pathology

2nd Edition

Editor: Lawrence Lacey
Paperback ISBN: 9780123868992
eBook ISBN: 9780123869005
Imprint: Academic Press
Published Date: 25th April 2012
Page Count: 504
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Manual of Techniques in Invertebrate Pathology, Second Edition, describes a wide range of techniques used in the identification, isolation, propagation/cultivation, bioassay, quantification, preservation, and storage of the major groups of entomopathogens, including entomophthorales, entomopathogenic fungi, entomopathogenic bacteria of the Bacilli, Nematode parasites, and pathogens and parasites of terrestrial molluscs. The book presents the perspectives of an international group of experts in the fields of invertebrate pathology, including microbiology, mycology, virology, nematology, biological control, and integrated pest management. Organized into 15 chapters, the book covers methods for the study of virtually every major group of entomopathogen, as well as methods for discovery and diagnosis of entomopathogens and the use of complementary methods for microscopy. It discusses the use of molecular techniques for identifying and determining phylogeny, factors that contribute to resistance to entomopathogens, and several other aspects of the science of invertebrate pathology. It also explains initial handling and diagnosis of diseased invertebrates, basic techniques in insect virology, and bioassay of bacterial entomopathogens against insect larvae. In addition, the reader is introduced to the use of bacteria against soil-inhabiting insects and preservation of entomopathogenic fungal cultures. The remaining chapters focus on research methods for entomopathogenic microsporidia and other protists, how the pathogenicity and infectivity of entomopathogens to mammals are tested, and preparations of entomopathogens and diseased specimens for more detailed study using microscopy. Experienced insect pathologists, biologists, entomologists, students, biotechnology personnel, technicians, those working in the biopesticide industry, and government regulators will find this manual extremely helpful.

Key Features

  • Step-by-step instructions for the latest techniques on how to isolate, identify, culture, bioassay and store the major groups of entomopathogens
  • New edition fully updated to address changes in the taxonomy of the vast majority of taxa
  • Discussion of safety testing of entomopathogens in mammals and also broader methods such as microscopy and molecular techniques
  • Provides extensive supplemental literature and recipes for media, fixatives and stains


Insect pathologists; biological control and integrated pest management specialists; entomologists; marine biologists, microbiologists; nematologists; protozoologists; virologists; mycologists and biologists involved in basic and applied research on invertebrate pathogens

Table of Contents



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



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



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





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



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


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



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




Chapter VIII. Mass production of entomopathogenic Hypocreales

1 Introduction

2 Culture maintenance

3 Solid Substrate Production of Aerial Conidia

4 Liquid Fermentation Production




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



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


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



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



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




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



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)






Color Plates


No. of pages:
© Academic Press 2012
25th April 2012
Academic Press
Paperback ISBN:
eBook ISBN:

About the Editor

Lawrence Lacey

Dr. Lacey earned his PhD in 1978 and has been internationally active working in medical entomology, insect pathology, and microbial control of economically and medically important insects. He conducted research on the biology and microbial control of dozens of insects in the US, Amazon Basin, West Africa in the Azores, throughout Europe, the Middle East and Western and Southeast Asia and the Pacific Northwest of the USA. In his 40 year career he has written or co-authored nearly 150 refereed papers in medical entomology, agricultural entomology, and insect pathology, and edited and co-edited seven books, including the Manual of Techniques in Invertebrate Pathology, second edition and Microbial Control of Insect and Mite Pests: From Theory to Practice.

Affiliations and Expertise

IP Consulting International, Yakima, WA, USA


listing in Society of Invertebrate Pathology Newsletter

Ratings and Reviews