Main description:

This book encompasses broad aspects of Cryptosporidium research with established methods that have been improved and expanded over the years as well as recent cutting-edge techniques. Within this collection are numerous molecular methods as well as protocols for genotyping and diagnostics, while also including room for in vitro cell culture techniques to address the issues with growing this difficult organism continuously. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Cryptosporidium: Methods and Protocols serves as an idea guide to the inherent challenges of working with cryptosporidial parasites to provide a foundation for new investigators to build upon.
The chapter "Accessing Cryptosporidium Omic and Isolate Data via CryptoDB.org" is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Contents:

1. Cryptosporidium Diagnostic Assays: Microscopy

Guy Robinson and Rachel M. Chalmers

2. Cryptosporidium Diagnostic Assays: Molecular Detection

Guy Robinson, Kristin Elwin, and Rachel M. Chalmers

3. Detection of Cryptosporidium Recovered From Large-Volume Water Samples Using Dead-End Ultrafiltration

Amy M. Kahler and Vincent R. Hill

4. Cryptosporidium Oocyst Purification Using Discontinuous Gradient Centrifugation

Michael J. Arrowood

5. Measuring Cryptosporidium Serologic Responses by Multiplex Bead Assay

Jeffrey W. Priest and Delynn M. Moss

6. Production and Purification of Functional Cryptosporidium Glycoproteins by Heterologous Expression in Toxoplasma gondii

Iwona Driskell and Roberta M. O'Connor

7. Cryptosporidium Genotyping for Epidemiology Tracking

Dawn M. Roellig and Lihua Xiao

8. Use of Markers to Determine Cryptosporidium Genotypes for Epidemiology Tracking and Detection

Anson V. Koehler and Jan Slapeta

9. Preparation of Cryptosporidium DNA for Whole Genome Sequencing

Guy Robinson and Rachel M. Chalmers

10. Accessing Cryptosporidium Omic and Isolate Data Via CryptoDB.org

Susanne Warrenfeltz and Jessica C. Kissinger on behalf of the EuPathDB Team

11. A Novel Method to Silence Genes in Cryptosporidium

Alejandro Castellanos-Gonzalez

12. Use of miRNAs to Study Host Cell-Parasite Interactions

Ji-Hao Xu, Nicholas W. Mathy, and Xian-Ming Chen

13. Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9

Adam Sateriale, Mattie Pawlowic, Sumiti Vinayak, Carrie Brooks, and Boris Striepen

14. Mouse Models for Use in Cryptosporidium Infection Studies and Quantification of Parasite Burden Using Flow Cytometry, qPCR, and Histopathology

Karine Sonzogni-Desautels, Jan R. Mead, and Momar Ndao

15. Calf Clinical Model of Cryptosporidiosis for Efficacy Evaluation of Therapeutics

Michael W. Riggs and Deborah A. Schaefer

16. Cell Culture Infectivity to Assess Chlorine Disinfection of Cryptosporidium Oocysts in Water

Jennifer L. Murphy and Michael J. Arrowood

17. High-Content Screening for Cryptosporidium Drug Discovery

Melissa S. Love and Case W. McNamara

18. High-Throughput Screening of Drugs Against the Growth of Cryptosporidium parvum In Vitro by qRT-PCR

Haili Zhang and Guan Zhu

19. In Vitro Culture of Cryptosporidium parvum Using Hollow Fiber Bioreactor: Applications for Simultaneous Pharmacokinetic and Pharmacodynamic Evaluation of Test Compounds

Nigel Yarlett, Mary Morada, Mohini Gobin, Wesley Van Voorhis, and Samuel Arnold

20. In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers

Georgia Wilke, Yi Wang, Soumya Ravindran, Thaddeus Stappenbeck, William Witola, and L. David Sibley

21. Two and Three Dimensional Bioengineered Human Intestinal Tissue Models for Cryptosporidium

Daviel Cardenas, Seema Bhalchandra, Hymlaire Lamisere, Ying Chen, Xi-Lei Zeng, Sasirekha Ramani, Umesh C. Karandikar, David L. Kaplan, Mary K. Estes, and Honorine D. Ward