Main description:

This detailed volume explores the field of bacterial virulence and the effort to understand how microbial interaction with a host results in the pathology of a specific disease. This collection of selected protocols includes advanced molecular biology and bioinformatics methods, cell culture and organoid models of infection, as well as in vivo infection models that are useful to study the interaction of pathogens with plants, insects, avian, and mammalian hosts. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and beneficial, Bacterial Virulence: Methods and Protocols serves as an ideal guide for researchers seeking to promote and further develop the exciting and continuously evolving field of bacterial virulence.
Chapter 19 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Contents:

Part I: Molecular Biology and Bioinformatics Methods

1. Generation of Markerless Gene Deletion Mutants in Listeria monocytogenes Using a Mutated PheS for Counterselection

Shai Ran Sapir, Etai Boichis, and Anat A. Herskovits

2. A Rapid Fluorescence-Based Screen to Identify Regulators and Components of Interbacterial Competition Mechanisms in Bacteria

Daniel Tchelet and Dor Salomon

3. Predicting Type III Effector Proteins Using the Effectidor Web-Server

Naama Wagner, Doron Teper, and Tal Pupko

4. Assay for Type III Secretion in Escherichia coli

Bosko Mitrovic and Neta Sal-Man

5. Profiling of Secreted Type 3 Secretion System Substrates by Salmonella enterica

Rivka Shem-Tov and Ohad Gal-Mor

Part II: Cell Culture and Organoid Models of Infection

6. Analysis of SPI-1 Dependent Type III Secretion and Injection Using a NanoLuc Luciferase-Based Assay

Sara Vilela Pais, Sibel Westerhausen, Erwin Bohn, and Samuel Wagner

7. Mycobacterium tuberculosis Infection of THP-1 Cells: A Model for High Content Analysis of Intracellular Growth and Drug Susceptibility

Leah Rankine-Wilson, Celine Rens, Henok Asfaw Sahile, and Yossef Av-Gay

8. Bone Marrow-Derived Macrophage (BMDM) Infection by Listeria monocytogenes

Etai Boichis, Shai Ran Sapir, and Anat A. Herskovits

9. Preparation and Inflammasome Activation of Murine Bone Marrow-Derived and Resident Peritoneal Macrophages

Izumi Sasaki and Tsuneyasu Kaisho

10. Flow Cytometry-Based Single Cell Analyses of Bacterial Adaption to Intracellular Environments

Marc Schulte and Michael Hensel

11. Quantification of Microbial Fluorescent Sensors during Live Intracellular Infections

Erez Mills and Erik Petersen

12. Dissecting Human Blood Immune Cells Response to Intracellular Infection Using Single Cell RNA-Sequencing

Shelly Hen-Avivi and Roi Avraham

13. Salmonella enterica Infection of Human and Mouse Colon Organoid-Derived Monolayers

Erin C. Boyle, Eva J. Wunschel, and Guntram A. Grassl

Part III: In Vivo Models of Infection

14. Analysis of Salmonella enterica Adhesion to Leaves of Corn Salad or Lettuce

Laura Elpers and Michael Hensel

15. Methods for Using the Galleria mellonella Invertebrate Model to Probe Enterococcus faecalis Pathogenicity

Ling Ning Lam, Debra N. Brunson, Jessica K. Kajfasz, and Jose A. Lemos

16. Murine Soft Tissue Infection Model to Study Group A Streptococcus (GAS) Pathogenesis in Necrotizing Fasciitis

Miriam Ravins, Poornima Ambalavanan, Debabrata Biswas, Rachel Tan Ying Min, Kimberly Lim Xuan Zhen, Yael Kaufman, Aparna Anand, Abhinay Sharma, and Emanuel Hanski

17. Mouse Model to Study Salmonella-Induced Colitis

Katrin Ehrhardt and Guntram A. Grassl

18. Analysis of Salmonella Typhi Pathogenesis in a Humanized Mouse Model

Taylor A. Stepien, Stephen J. Libby, Joyce E. Karlinsey, Michael A. Brehm, Dale L. Greiner, Leonard D. Shultz, Thea Brabb, and Ferric C. Fang

19. In Vivo Tracking of Bacterial Colonization in Different Murine Models Using Bioluminescence: The Example of Salmonella

Michael Koczerka, Isabelle Lantier, Anne Pinard, Marie Morillon, Justine Deperne, Ohad Gal-Mor, Olivier Grepinet, and Isabelle Virlogeux-Payant

20. Two In Vivo Models to Study Salmonella Asymptomatic Carrier State in Chicks

Philippe Velge, Pierrette Menanteau, Thierry Chaumeil, Emilie Barilleau, Jerome Trotereau, and Isabelle Virlogeux-Payant