Short description:

This book is a compilation of protocols for conducting experiments with a variety of yeasts, including S. cerevisiae, Candida albicans, non-albicans Candida sp., Cryptococcus sp.; and molds, including Aspergillus sp., Fusarium sp., Mucor, Rhizopus. The chapters not only discuss National Committee for Clinical Laboratory Standards (NCCLS) accredited protocols but also emphasize the significance of Biosafety Level-2 (BSL-2) facilities. Subsequent chapters cover the essentials of handling, culturing, preserving, and reviving various fungal pathogens. Moreover, advanced techniques are explored, including counting colony-forming units (CFU), growth curve preparation, and microtiter plate assays. A separate chapter discusses the protocols for disc-diffusion, spot and kill-kinetics assays, addressing analyses like a checkerboard and Fractional Inhibitory Concentration (FIC) index assessments. Additionally, the book covers the formation of biofilm, capsule and melanin, fungal cell adhesion, and sterols extraction. Finally, it elaborates on the optimized impact achieved by assessing mitochondrial fitness through non-fermentable carbon.

This book is relevant for mycologists and scientists working in the field, providing insights into fungal pathogens and protocols for handling different fungal species.

Long description:

This book is a compilation of protocols for conducting experiments with a variety of yeasts, including S. cerevisiae, Candida albicans, non-albicans Candida sp., Cryptococcus sp.; and molds, including Aspergillus sp., Fusarium sp., Mucor, Rhizopus. The chapters not only discuss National Committee for Clinical Laboratory Standards (NCCLS) accredited protocols but also emphasize the significance of Biosafety Level-2 (BSL-2) facilities. Subsequent chapters cover the essentials of handling, culturing, preserving, and reviving various fungal pathogens. Moreover, advanced techniques are explored, including counting colony-forming units (CFU), growth curve preparation, and microtiter plate assays. A separate chapter discusses the protocols for disc-diffusion, spot and kill-kinetics assays, addressing analyses like a checkerboard and Fractional Inhibitory Concentration (FIC) index assessments. Additionally, the book covers the formation of biofilm, capsule and melanin, fungal cell adhesion, and sterols extraction. Finally, it elaborates on the optimized impact achieved by assessing mitochondrial fitness through non-fermentable carbon.

This book is relevant for mycologists and scientists working in the field, providing insights into fungal pathogens and protocols for handling different fungal species.

Table of Contents:

1. Introduction to BSL-2 Facilities.- 2. Handling, Culturing, Preservation and Revival of Fungal Pathogens.- 3. Counting Colony Forming Units (CFU) to Estimate Viable Fungal Cells in a Sample.- 4. Growth Curve Preparation for the Calculation of Fungal Cell Count Over Time.- 5. Microtiter Assay for Drug Susceptibility Testing of Pathogenic Fungi in Broth.- 6. Disk Diffusion Assay for Testing the Drug Sensitivity of Pathogenic Fungi in Agar.- 7. Spot Assay for Comparing Drug Susceptibility of Pathogenic Fungi in Agar.- 8. Time-Kill Kinetics to Determine the Number of Living Cells Over a Period of Time.- 9. Checkerboard Analysis to Calculate the Fractional Inhibitory Concentration (FIC) Index of Pathogenic Fungi for Drug Interaction.- 10. Hyphal Induction in Yeasts and Molds.- 11. Isolation of Genomic DNA from Pathogenic Fungi.- 12. Exopolysaccharide Secretion by Pathogenic Fungi.- 13. Biofilm Formation Test and Determination of the Effect of Drugs on their Formation.- 14. Induction of Capsule Formation in Cryptococcus sp.- 15. Melanin Formation in Cryptococcus sp.- 16. Urease Assay for Cryptococcus sp.- 17. Phospholipase Assay for the Detection of Virulence of Pathogenic Fungi.- 18. Hydrophobicity Test for Virulence of Pathogenic Fungi.- 19. Adhesion of Fungal Cells to Buccal Epithelial Cells.- 20. Sterols Extraction, Analysis and Quantification in Pathogenic Fungi.- 21. Assessing Mitochondrial Fitness by Assessing Microbial Growth on Non-Fermentable Carbon Sources.- 22. In Vivo Assay using Caenorhabditis elegans.