Main description:

This volume looks at all aspects of manipulation of Leptospira spp. from strain isolation to the latest techniques used to study the pathogenesis of leptospirosis. The chapters in this book cover topics such as the procedure to cultivate and isolate leptospires from both clinical and environmental samples; using methods like whole genome sequencing and Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry to identify bacterial species; tools for gene inactivation and in vitro and in vivo assays to study the pathogenesis of leptospirosis; and the use of hamsters to evaluate leptospiral virulence and vaccine candidates. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Leptospira spp.: Methods and Protocols is a valuable resource for researchers interested in learning more about this developing field and these fascinating organisms.

Contents:

Preface...
Table of Contents...
Contributing Authors...

1. Isolation and Culture of Leptospira from Clinical and Environmental Samples
Cyrille Goarant, Dominique Girault, Roman Thibeaux, and Marie-Estelle Soupe-Gilbert

2. Core Genome Multi-Locus Sequence Typing Analyses of Leptospira spp. using the Bacterial Isolate Genome Sequence Database
Linda Grillova and Mathieu Picardeau

3. Use of MALDI-ToF Mass Spectrometry for Identification of Leptospira
Dominique Girault, Malia Kainiu, Emilie Barsac, Roman Thibeaux, and Cyrille Goarant

4. Cell Enumeration of Leptospira by Flow Cytometry
Celia Fontana and Jean Reyes

5. The Single-Step Method of RNA Purification Applied to Leptospira
Crispin Zavala-Alvarado and Nadia Benaroudj

6. Purification of LPS from Leptospira
Delphine Bonhomme and Catherine Werts

7. Isolation, Purification, and Characterization of Leptophages
Olivier Schiettekatte and Pascale Bourhy

8. Creating a Library of Random Transposon Mutants in Leptospira
Christopher J. Pappas, Hui Xu, and Md A. Motaleb

9. Transposon Sequencing in Leptospira interrogans
Kristel Lourdault and James Matsunaga

10. Specific Gene Silencing in Leptospira biflexa by RNA-Guided Catalytically inactive Cas9 (dCas9)
Luis Guilherme Virgilio Fernandes and Ana Lucia Tabet Oller Nascimento

11. Leptospira spp. Toolbox for Chemotaxis Assay
Ambroise Lambert

12. In Situ Structural Analysis of Leptospira spp. by Electron Cryotomography
Akihiro Kawamoto

13. Measurement of the Cell-Body Rotation of Leptospira
Shuichi Nakamura

14. In Vivo Imaging of Bioluminescent Leptospires
Frederique Vernel-Pauillac and Catherine Werts

15. Cell Monolayer Translocation Assay
Elsio A. Wunder, Jr.

16. Cell Adhesion Assay to Study Leptospiral Proteins: An Approach to Investigate Host-Pathogen Interaction
Aline F. Teixeira and Ana L. T. O. Nascimento

17. Complement Resistance Assays
Lourdes Isaac and Angela Silva Barbosa

18. Evaluation of Intracellular Trafficking in Macrophages
Claudia Toma and Toshihiko Suzuki

19. Biofilm Formation and Quantification using the 96-Microtiter Plate
Roman Thibeaux, Malia Kainiu, and Cyrille Goarant

20. Survival Tests for Leptospira spp.
Clemence Mouville and Nadia Benaroudj

21. Cultivation of Leptospira interrogans within Rat Peritoneal Dialysis Membrane Chambers
Andre Alex Grassmann and Melissa J. Caimano

22. Use of Golden Syrian Hamster as an Animal Model to Study Leptospirosis-Associated Immune Responses
Julie Cagliero, Karl Huet, and Mariko Matsui

23. Evaluation of Vaccine Candidates against Leptospirosis using Golden Syrian Hamsters
Karen V. Evangelista and Kristel Lourdault

24. Leptospira and Leptospirosis
Mathieu Picardeau

25. Laboratory Diagnosis of Leptospirosis
Nobuo Koizumi

Subject Index List...