Leptospira spp.

This detailed new edition reflects the rapid evolution of Leptospira research over the past decade, particularly in genetics, genomics, imaging, and computational analysis. The book provides practical guidance for the manipulation of Leptospira spp. and for the investigation of leptospiral pathogenesis using state-of-the-art experimental approaches by introducing alternative media for direct isolation from infected hosts and for better mimicking host-associated conditions and by exploring gene editing tools, imaging-based approaches, and host-pathogen interactions, among other subject areas. 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 and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Leptospira spp. : Methods and Protocols, Second Edition serves both established investigators and newcomers to the field, facilitating rigorous and innovative research on these fastidious yet fascinating organisms.

Isolation and Culture of Leptospira from Clinical and Environmental Samples. - Preparation of HAN Medium for the Isolation of Leptospira Directly from Infected Hosts. - EMEM and DMEM as an Alternative Media for Leptospiral Culture, Diagnostic, and Pathogenesis Studies. - High Resolution Genotyping of Leptospira Direct from Clinical Samples Using Multiplexed Amplicon Sequencing. - Core Genome Multi-Locus Sequence Typing Analyses of Leptospira spp. Using the Bacterial Isolate Genome Sequence Database. - Use of MALDI-ToF Mass Spectrometry for Identification of Leptospira. - The Single-Step Method of RNA Purification Applied to Leptospira. - Purification of Lipopolysaccharides from Leptospira. - Isolation, Purification, and Characterization of Leptophages. - Creating a Library of Random Transposon Mutants in Leptospira. - Transposon Sequencing in Leptospira interrogans. - Gene Knockout in Leptospira spp. by CRISPR/Cas9-NHEJ (Non-Homologous End-Joining) and CRISPR-Prime Editing. - Leptospira spp. Toolbox for Chemotaxis Assay. - In Situ Structural Analysis of Leptospira spp. by Electron Cryotomography. - Bacterial Size Analysis Using High-Content Imaging: Application to Leptospira Strains. - Measurement of the Cell-Body Rotation of Leptospira. - Machine Learning-Based, Label-Free Motion Tracking of Leptospira spp. on Cultured Cells. - Machine Learning-Based Quantification of Motility Parameters in Individual Leptospira Cells. - Instance Segmentation as a Foundation for Quantitative Image Analysis in Leptospira Research. - In Vivo Imaging of Bioluminescent Leptospires. - Cell Monolayer Translocation Assay. - Cell Adhesion Assays to Investigate Leptospiral Protein Interactions with ECM, Cell Receptors, and Cultured Mammalian Cells: Methods for Understanding Host-Leptospira Interactions. - Complement Resistance Assays. - Evaluation of Intracellular Trafficking in Macrophages. - Protocols for Standard and Advanced Visualization and Quantification of Leptospira Biofilms. - Hamster Model of Leptospira Infection. - Cultivation of Leptospira interrogans Within Rat Peritoneal Dialysis Membrane Chambers. - Determination of Bacterial Adherence to Mouse Tissues Using a Hematogenous Model. - Leptospira and Leptospirosis. - Laboratory Diagnosis of Leptospirosis.