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INTRODUCTION

Cultivation is the process of propagating organisms by providing the proper environmental conditions. Parasites, bacteria, and viruses all generally require cultivation for detailed study. The field of microbiology has the greatest experience in the cultivation of bacteria and as such, this is the focus of this chapter.

Bacteria divide by binary fission, asexual reproduction where a single cell divides giving rise to two cells. Those two cells give rise to a total of four cells and so on. This process of replication requires the acquisition of elements that make up their chemical composition. Nutrients from the environment provide these elements in metabolically accessible forms. In addition, organisms require metabolic energy to synthesize macromolecules and maintain essential chemical gradients across their membranes. Factors that must be controlled during growth include the nutrients, pH, temperature, aeration, salt concentration, and ionic strength of the medium.

REQUIREMENTS FOR GROWTH

Most of the dry weight of microorganisms is organic matter containing the elements carbon, hydrogen, nitrogen, oxygen, phosphorus, and sulfur. In addition, inorganic ions such as potassium, sodium, iron, magnesium, calcium, and chloride are required to facilitate enzymatic catalysis and to maintain chemical gradients across the cell membrane.

For the most part, the organic matter is in macromolecules formed by the introduction of anhydride bonds between building blocks. Synthesis of the anhydride bonds requires chemical energy, which is provided by the two phosphodiester bonds in adenosine triphosphate (ATP; see Chapter 6). Additional energy required to maintain a relatively constant cytoplasmic composition during growth in a range of extracellular chemical environments is derived from the proton motive force. The proton motive force is the potential energy that can be derived by passage of a proton across a membrane. In eukaryotes, the membrane may be part of the mitochondrion or the chloroplast. In prokaryotes, the membrane is the cytoplasmic membrane of the cell.

The proton motive force is an electrochemical gradient with two components: a difference in pH (hydrogen ion concentration) and a difference in ionic charge. The charge on the outside of the bacterial membrane is more positive than the charge on the inside, and the difference in charge contributes to the free energy released when a proton enters the cytoplasm from outside the membrane. Metabolic processes that generate the proton motive force are discussed in Chapter 6. The free energy may be used to move the cell, to maintain ionic or molecular gradients across the membrane, to synthesize anhydride bonds in ATP, or for a combination of these purposes. Alternatively, cells given a source of ATP may use its anhydride bond energy to create the proton motive force that in turn may be used to move the cell and to maintain chemical gradients.

To grow, a bacterium requires all the elements in its organic matter as well as the full complement of ions required ...

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