Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
ix
1 Isolation and Culture of Leptospira from Clinical
and Environmental Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
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 . . . . . 11
Linda Grillova ´ and Mathieu Picardeau
3 Use of MALDI-ToF Mass Spectrometry for Identification
of Leptospira. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Dominique Girault, Malia Kainiu, Emilie Barsac,
Roman Thibeaux, and Cyrille Goarant
4 Cell Enumeration of Leptospira by Flow Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . 31
Ce ´lia Fontana and Jean Reyes
5 The Single-Step Method of RNA Purification Applied to Leptospira . . . . . . . . . . . 41
Crispin Zavala-Alvarado and Nadia Benaroudj
6 Purification of LPS from Leptospira . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Delphine Bonhomme and Catherine Werts
7 Isolation, Purification, and Characterization of Leptophages . . . . . . . . . . . . . . . . . 67
Olivier Schiettekatte and Pascale Bourhy
8 Creating a Library of Random Transposon Mutants in Leptospira . . . . . . . . . . . . . 77
Christopher J. Pappas, Hui Xu, and Md A. Motaleb
9 Transposon Sequencing in Leptospira interrogans . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
Kristel Lourdault and James Matsunaga
10 Specific Gene Silencing in Leptospira biflexa by RNA-Guided
Catalytically Inactive Cas9 (dCas9). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Luis Guilherme Virgı ´lio Fernandes,
and Ana Lucia Tabet Oller Nascimento
11 Leptospira spp. Toolbox for Chemotaxis Assay. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
Ambroise Lambert
12 In Situ Structural Analysis of Leptospira spp. by Electron
Cryotomography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Akihiro Kawamoto
13 Measurement of the Cell-Body Rotation of Leptospira. . . . . . . . . . . . . . . . . . . . . . . 139
Shuichi Nakamura
14 In Vivo Imaging of Bioluminescent Leptospires . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
Fre ´de ´rique Vernel-Pauillac and Catherine Werts
15 Cell Monolayer Translocation Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Elsio A. Wunder Jr.
vii
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
ix
1 Isolation and Culture of Leptospira from Clinical
and Environmental Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1
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 . . . . . 11
Linda Grillova ´ and Mathieu Picardeau
3 Use of MALDI-ToF Mass Spectrometry for Identification
of Leptospira. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Dominique Girault, Malia Kainiu, Emilie Barsac,
Roman Thibeaux, and Cyrille Goarant
4 Cell Enumeration of Leptospira by Flow Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . 31
Ce ´lia Fontana and Jean Reyes
5 The Single-Step Method of RNA Purification Applied to Leptospira . . . . . . . . . . . 41
Crispin Zavala-Alvarado and Nadia Benaroudj
6 Purification of LPS from Leptospira . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Delphine Bonhomme and Catherine Werts
7 Isolation, Purification, and Characterization of Leptophages . . . . . . . . . . . . . . . . . 67
Olivier Schiettekatte and Pascale Bourhy
8 Creating a Library of Random Transposon Mutants in Leptospira . . . . . . . . . . . . . 77
Christopher J. Pappas, Hui Xu, and Md A. Motaleb
9 Transposon Sequencing in Leptospira interrogans . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
Kristel Lourdault and James Matsunaga
10 Specific Gene Silencing in Leptospira biflexa by RNA-Guided
Catalytically Inactive Cas9 (dCas9). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Luis Guilherme Virgı ´lio Fernandes,
and Ana Lucia Tabet Oller Nascimento
11 Leptospira spp. Toolbox for Chemotaxis Assay. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
Ambroise Lambert
12 In Situ Structural Analysis of Leptospira spp. by Electron
Cryotomography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Akihiro Kawamoto
13 Measurement of the Cell-Body Rotation of Leptospira. . . . . . . . . . . . . . . . . . . . . . . 139
Shuichi Nakamura
14 In Vivo Imaging of Bioluminescent Leptospires . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
Fre ´de ´rique Vernel-Pauillac and Catherine Werts
15 Cell Monolayer Translocation Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Elsio A. Wunder Jr.
vii