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Chapter 47 : Culture Preservation

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Culture Preservation, Page 1 of 2

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Abstract:

The primary aim of culture preservation is to maintain the organism alive, uncontaminated, and without variation or mutation, that is, to preserve the culture in a condition that is as close as possible to the original isolate. The major disadvantages of the serial-transfer technique are the risks of contamination, transposition of strain numbers or designations (mislabeling), selection of variants or mutants, and possible loss of culture, as well as the required storage space. Commercially available strips impregnated with spores are used to check the efficacy of the sterilization equipment and cycle. Many heterotrophic bacteria can be preserved in dried gelatin drops or disk. A variety of bacteria have been successfully preserved by drying on sterile silica gel granules. Freeze-drying (lyophilization) is one of the most economical and effective methods for long-term preservation of bacteria and other microorganisms. Two of the most common methods used are centrifugal freeze-drying and prefreezing. Furthermore, many culture collections serve as centers of expertise for preserving the microbial germplasm and are very useful in training others in the do's and don't's of culture preservation and maintenance.

Citation: Gherna R, Reddy C. 2007. Culture Preservation, p 1019-1034. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch47

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Figures

Image of FIGURE 1
FIGURE 1

Double-vial method of freeze-drying. (A) Vacuum gauge; (B) vacuum pump; (C) VirTis condenser; (D) reservoir filled with dry ice and ethylene glycol; (E) acrylic top plate; (F) stainless steel pan filled with crushed dry ice and ethylene glycol; (G) specimen vial.

Citation: Gherna R, Reddy C. 2007. Culture Preservation, p 1019-1034. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch47
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Image of FIGURE 2
FIGURE 2

Manifold method of freeze-drying. (A) Vacuum gauge; (B) vacuum pump; (C) VirTis condenser; (D) reservoir filled with dry ice and ethylene glycol; (E) specimen vial; (F) stainless steel pan filled with crushed dry ice and ethylene glycol; (G) manifold.

Citation: Gherna R, Reddy C. 2007. Culture Preservation, p 1019-1034. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch47
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Image of FIGURE 3
FIGURE 3

Sectional view (top) of acrylic top plate, chamber double-vial method. (A) Vacuum release valve; (B) acrylic plastic top (7 in. wide by 7 in. long by 1 in. deep); (C) stainless steel inlet port (outer diameter, 11/16 in.; inner diameter, 1/2 in.); (D) hole (inner diameter, 1/2 in.) drilled in acrylic plastic to accept the stainless steel tube; (E) rubber gasket (7 in. wide by 7 in. long by 1/4 in. deep) with 2 3/8-in. hole drilled in the center; (F) center hole in bottom of acrylic plastic plate (diameter, 0.5 in.); (G) stainless steel specimen pan (5 in. long by 5 in. wide by 4 in. deep). Top view (bottom) of acrylic top plate, showing Tygon pressure tubing attached to the stainless steel tube and the pressure release valve on top.

Citation: Gherna R, Reddy C. 2007. Culture Preservation, p 1019-1034. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch47
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Image of FIGURE 4
FIGURE 4

Opening of chamber double vials and manifold single vials. (Courtesy of the ATCC.)

Citation: Gherna R, Reddy C. 2007. Culture Preservation, p 1019-1034. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch47
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References

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Tables

Generic image for table
TABLE 1

Expected shelf life of 33 representative bacterial genera preserved by various methods

The transfer schedule depends on the medium used. The times listed are approximations, and species variation occurs within a genus.

Depends on species.

Spirillum volutans.

Citation: Gherna R, Reddy C. 2007. Culture Preservation, p 1019-1034. In Reddy C, Beveridge T, Breznak J, Marzluf G, Schmidt T, Snyder L (ed), Methods for General and Molecular Microbiology, Third Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817497.ch47

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