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Chapter 12 : Culture Techniques

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Abstract:

This chapter talks about the main culture techniques for microbial growth. It first describes the solid, semisolid, biphasic, membrane surface, and immobilized culture techniques. This is followed by laboratory scale liquid culture techniques including specialized liquid cultures such as synchronous and dialysis cultures. Solid media are also used in mass culture, bioautography, and physiological studies of bacterial cells. Solid culture is one of the most useful techniques in the isolation and cultivation from single cells. The solid surface usually is that of an agar or otherwise solidified medium. Semisolid media are also useful in chemotaxis studies. For example, a semisolid medium containing an oxidizable carbon and energy source can be used to investigate positive chemotaxis in , , , and many other species. Laboratory scale liquid cultures provide one of the most common techniques to grow and study the behavior of microorganisms. The section on energetics and stoichiometry describes the theoretical aspects of microbial growth focusing on the electron acceptor and donor and the carbon and nitrogen sources. There are two basic principles underlying dialysis culture. First, it provides a means for achieving substrate limited growth. Second, dialysis culture provides a means for lowering the concentration of a diffusible metabolite product inhibitory to growth; the product in the culture chamber diffuses through the membrane and is diluted in the larger dialysate reservoir, thus relieving the feedback inhibition by the product that normally regulates its production.

Citation: Hashsham S. 2007. Culture Techniques , p 270-285. 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.ch12

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FIGURE 1

Thick glass culture tube with butyl rubber stoppers and aluminum crimp caps useful in isolation and culturing of anaerobic microorganisms.

Citation: Hashsham S. 2007. Culture Techniques , p 270-285. 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.ch12
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Image of FIGURE 2
FIGURE 2

Schematic of an anaerobic continuous stirred tank reactor with separate pumps for supplying substrate and nutrients, effluent pump with water seal, and gas collection line. For smaller systems with multiple reactors, syringe pumps with multiple ports can be used.

Citation: Hashsham S. 2007. Culture Techniques , p 270-285. 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.ch12
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Tables

Generic image for table
TABLE 1

Electron acceptors and donors and carbon sources for selected microorganisms

TCE, trichloroethene.

PCE, tetrachloroethene.

Citation: Hashsham S. 2007. Culture Techniques , p 270-285. 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.ch12

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