Bacterial biofilms in nature and disease.
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Biomedical subjects
Publications and source records attributed to G G Geesey.
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Copper-binding activity by exopolymers from adherent cells of a freshwater-sediment bacterium was demonstrated by a combination of equilibrium dialysis and flameless atomic absorption spectrometry. Crude, cell-free exopolymer preparations containing protein and polysaccharide components bound up to 37 nmol of Cu per mg (dry weight). A highly purified exopolysaccharide preparation bound up to 253 nmol of Cu per mg of carbohydrate. The conditional stability constant for the crude exopolymer-Cu complex was 7.3 X 10(8). This value was similar to those obtained for Cu complexes formed with humic acids and xanthan, an exopolysaccharide produced by Xanthomonas campestris. Studies conducted at copper concentrations, pHs, and temperatures found in sediments from which the bacterium was isolated indicated that the exopolymers were capable of binding copper under natural conditions.
Cells of a freshwater sediment bacterium produced firmly bound extracellular polymers in laboratory cultures which, at the ultrastructural level, resembled those produced by natural sediment bacterial populations. Production of the exopolymers during subculture was maintained by using as a source of inoculum the population of cells which adhered to each other and to the wall of the glass culture vessel. The exopolymers were selectively released from the cells by blending and centrifugation in the presence of EDTA. Evaluation of glucose-6-phosphate dehydrogenase activity and 2-keto-3-deoxyoctonate concentration indicated that only small amounts of intracellular and cell wall components were released from the cells during exopolymer removal. Chemical analysis of the isolated crude exopolymer material indicated that it contained protein, polysaccharide, and DNA. The treatment promoted the selective isolation of firmly bound polymers from the surface of adherent cells.
Cells of a psychrophilic marine bacterium were found to take up a variety of amino acids from seawater. Some of the amino acids that were taken up were released when the cells were exposed to a hypotonic salt solution. The proportion that was released varied according to the amino acid. A pool of the amino acid arginine that was formed during very short periods of exposure of cells to the exogenously supplied amino acid was particularly sensitive to reductions in salinity. In general, exposure to hypotonic salt solutions also resulted in reduced amino acid uptake by the cells. Complete removal of seawater salts (SE treatment) produced obvious structural alterations in the cell envelope, resulting in an even greater reduction in amino acid uptake. Under these conditions, amino acid-binding components were released by the cells. Differential centrifugation and fluorescent antibody studies indicated that arginine-binding components are located on or near the surface of intact cells. The data suggest that substrate receptors were sensitive to reductions in seawater salt concentrations and that lesions at this level affected the organism's substrate uptake and retention capabilities.
The cells of the marine bacterium Ant-300 were found to take up arginine when this substrate was at low concentrations. The cells possessed an uptake system(s) that specifically transported l-arginine. The kinetic parameters for uptake appeared to differ when the cells were exposed to nanomolar and micromolar concentrations of the amino acid. Uptake over this concentration range functioned in the absence of an exogenous energy source, even after the cells had been preincubated in unsupplemented artificial seawater. Respiratory activity appeared to be a more important driving force for arginine uptake than adenosine 5'-triphosphate hydrolysis. The cells also exhibited chemotaxis toward l-arginine. The minimum arginine concentration needed to elicit a chemotactic response was between 10 and 10 M. It is proposed that the capture of arginine by cells of Ant-300 in nutrient-depleted waters, which are typical of the open ocean, proceeds via high-affinity active transport, whereas in substrate-enriched seawater, capture involves chemotaxis and an active transport mechanism with reduced affinity for the substrate.
Epifluorescent microscopy showed as many as 4 x 10(6) bacteria/mL in the turbid waters of the Athabasca River near the tar sand deposits in northeastern Alberta. The numbers were usually similar upstream and downstream (60 km) from pilot-mining operations. The majority of bacteria existed as free-living cells in spite of the fact there were high concentrations of suspended sediment present (average 220 mg/L) during the ice-free period. Fluctuations in bacterial concentration were positively correlated (r = 0.86, P less than 0.05) with total organic carbon concentrations in the river water.
Natural populations of bacteria assoiciated with the slime on submerged surfaces in a mountain stream were examined by phase-contrast and electron microscopy. The slime contained large numbers of bacteria which were predominantly gram-negative as determined by their cell wall structure. Examination of the in situ distribution of cells revealed that they were enmeshed in an extensive fibrous matrix whose component fibrils were stained with ruthenium red. The arrangement of slime fibrils immediately around individual bacterial cells suggested that this material was produced by these bacteria. This slime facilitated microcolony development and also anchored the bacteria to a particular surface. It is proposed that these slime-enmeshed microcolonies constitute functional communities within which most sessile bacteria live.
The heat inactivation of the obligately psychrophilic marine bacterium Ant-300 was investigated in terms of glucose uptake, the oxidation of glucose to CO2, and permeability control. At 13C, the maximum temperature for growth, and at slightly higher temperatures, CO2 evolution decreased with time during the oxidation of exogenously supplied glucose. The decrease in CO2 evolution appeared to be a result of heat-induced restrictions on glucose uptake. Leakage of intracellular metabolites apparently contributed to the cells decreased ability to take up glucose at elevated temperatures. A consequence of these heat-induced changes seemed to be the acceleration of cell starvation.