Title: Preservation of entomopathogenic fungal cultures Author
Submitted to: Manual of Techniques in Invertebrate Pathology
Publication Type: Book / Chapter
Publication Acceptance Date: January 19, 2012
Publication Date: April 1, 2012
Citation: Humber, R.A. 2012. Preservation of entomopathogenic fungal cultures. In: Lacey, L.A., editor. Manual of Techniques in Invertebrate Pathology. Chapter 10. London, UK: Academic Press. p. 317-328. Technical Abstract: There are many possible techniques that are widely used for the long-term preservation of fungal cultures. This chapter focusses on the fungi that are pathogenic to insects and other invertebrate hosts (and which provide more challenges for preservation than most fungal saprobes) while providing means to evaluate the most technologically appropriate and cost-effective method for any given laboratory to use to meet their needs for preserving fungal cultures. Step-by-step protocols are given for the preservation (and recovery from preservation) for the most routinely used methods which are organized by the temperatures at which preservation is done: Three methods (serial transfer, preservation under mineral oil, and immersion in distilled water) are described from no frozen storage. Freeze-drying techniques that are almost universally satisfactory are described, as well as the fundamental differences between fungi that will or will not tolerate freeze-drying. Storage techniques in freezers are broken into those that may tolerate freezing in standard appliance freezers (at ca. -20 deg C), and cryogenic techniques in ultracold freezers (at -80 and -120 deg C), and in liquid nitrogen dewars (in either vapor- or liquid-phase storage) at temperatures ranging downwards from ca. -120 deg C to -196 deg C. There is also a discussion of the recordkeeping that needs to be done for long-term culture storage, and of techniques that are not widely used but that may be excellent choices in some laboratories for specialized culture preservation needs.