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United States Department of Agriculture

Agricultural Research Service

Title: Cultivation of Spiroplasma in Undefined and Defined Media

Authors
item HACKETT, KEVIN
item Lynn, Dwight

Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: June 30, 1995
Publication Date: N/A

Interpretive Summary: Spiroplasmas are helical, motile bacteria that cause disease in insects, ticks, and plants. Because spiroplasmas differ in their ability to grow in growth medium, strategies for their cultivation have varied, requiring specialized media. Certain fast-growing spiroplasmas and one fastidious spiroplasma grow in chemically defined media (ones in which all the chemicals are known), facilitating nutritional analysis and development of mutants for molecular studies. Spiroplasma media have been modified not only according to the target organism but also for their intended application. Whereas media used for initial isolation usually contain a rich mixture of nutrients, many fast growing, insect-associated spiroplasmas can be routinely maintained in simplified undefined media. The current chapter describes systems for cultivating both nutritionally fastidious spiroplasmas, and easy to grow ones. Principles developed for cultivating fastidious spiroplasmas should also have application to those attempting to cultivate closely related human and veterinary mycoplasmas.

Technical Abstract: Spiroplasmas are helical, motile bacteria that cause disease in insects, ticks, and plants. Because spiroplasmas differ in their ability to grow in medium formulations, strategies for their cultivation have varied, requiring specialized media. Certain fast- growing spiroplasmas and one fastidious spiroplasma grow in chemically defined media, facilitating nutritional analysis and development of mutants for molecular studies. Spiroplasma media have been modified not only according to the target organism but also for their intended application. Whereas media used for primary isolation usually contain a rich mixture of nutrients, many fast growing, insect-associated spiroplasmas can be routinely maintained in simplified undefined media. The current chapter describes systems for cultivating both fastidious spiroplasmas, and easy to grow ones. General cultivation principles, developed in the chapter, include: (i) amino acids (ii) or bovine serum albumin are important components. (iii) More fastidious spiroplasmas may be cultured under anaerobic conditions or in cell co- culture. (iv) Serum batch, or lipid mixture (for defined media), can be critical. (v) Inorganic salts are probably unimportant, within reasonable physiological concentrations. (vi) Trace amounts of some components may occur as contaminants in medium components or glassware. (vii) Organic acids are not required, but may enhance growth of some spiroplasmas. (viii) Media made with a combination of adenosine, guanosine, cytidine, and thymidine support better growth than those with fewer nucleosides.

Last Modified: 9/10/2014
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