Alterations in the composition and bacteriophage-binding properties of walls of Staphylococcus aureus H grown in continuous culture in simplified defined media

A nutritional mutant of Staphylococcus aureus H has been isolated and grown in media in which the only amino acids are arginine, cysteine, glutamic acid and proline. Walls of the bacteria grown in such media in continuous culture under potassium limitation differ in composition from walls of the bacteria grown in batch culture in rich nutrient broth in that they contain less glycine, the peptidoglycan component is less highly cross-linked and the teichoic acid component contains a reduced proportion of $\text{N-}\text{acetylglucosaminyl}$ substituents. Walls of the potassium-limited bacteria retain the ability to bind bacteriophage 52a but are more susceptible to the action of lytic peptidases than are wall samples in which the peptidoglycan is more highly cross-linked. Teichoic acid was present in walls of the bacteria grown under phosphate limitation in the defined medium and these walls were also able to absorb bacteriophage 52a.     

Effect of dilution rate and Mg2+ limitation on toxic shock syndrome toxin-1 production by Staphylococcus aureus grown in defined continuous culture

A toxic shock syndrome isolate of Staphylococcus aureus was grown in a chemostat, in a defined synthetic medium of six amino acids, glucose, two vitamins and salts. Steady states were achieved under limiting and replete Mg2+ conditions and at a range of relative specific growth rates. The biomass and toxic shock syndrome toxin-1 (TSST-1) were estimated at each condition. Under Mg2+ limitation the biomass and TSST-1 production rates were reduced compared to Mg2+ replete conditions. Optimal TSST-1 production occurred at 0.81 relative specific growth rate.     

Evaluation of culture media for recovery of Staphylococcus aureus from swimming pools.

Several selective media were evaluated for the primary isolation and enumeration of Staphylococcus aureus from halogenated indoor swimming pool waters. Standard plate counts of the viable population and total coliform densities were also determined to ascertain their value as indicator systems. All studies were done with membrane filters. The most selective, accurate, and reliable medium was Vogel-Johnson (VJ) medium supplemented with 0.5% pyruvate. This medium recovered two times more typical colonies than VJ medium alone, and subsequent identification of these well-defined black colonies proved that approximately 80% were S. aureus. The S. aureus recoveries correlated well with halogen levels and bather density use also. In contrast, VJ medium alone was 60% selective for S. aureus, and VJ medium supplemented with catalase did not increase either the percent recovery or the selectivity over that of VJ medium alone. Standard plate counts did not correlate with halogen levels, bather density, or total viable colonies. Coliforms were rarely recovered from indoor pool waters and were not considered to be useful indicators of water quality.     

Evaluation of culture media for recovery of Staphylococcus aureus from swimming pools

Several selective media were evaluated for the primary isolation and enumeration of Staphylococcus aureus from halogenated indoor swimming pool waters. Standard plate counts of the viable population and total coliform densities were also determined to ascertain their value as indicator systems. All studies were done with membrane filters. The most selective, accurate, and reliable medium was Vogel-Johnson (VJ) medium supplemented with 0.5% pyruvate. This medium recovered two times more typical colonies than VJ medium alone, and subsequent identification of these well-defined black colonies proved that approximately 80% were S. aureus. The S. aureus recoveries correlated well with halogen levels and bather density use also. In contrast, VJ medium alone was 60% selective for S. aureus, and VJ medium supplemented with catalase did not increase either the percent recovery or the selectivity over that of VJ medium alone. Standard plate counts did not correlate with halogen levels, bather density, or total viable colonies. Coliforms were rarely recovered from indoor pool waters and were not considered to be useful indicators of water quality.     

Comparative study of the growth of Listeria monocytogenes in defined media and demonstration of growth in continuous culture

C.E. JONES, G. SHAMA, P.W. ANDREW, I.S. ROBERTS AND D.JONES. 1995 A basic requirement for physiological studies with Listeria monocytogenes is a chemically defined medium that supports growth of the bacterium in batch and continuous culture. A number of such media have been devised but comparative studies of their efficiency are few and none has been used in continuous culture. Six of the media were compared for their ability to sustain sequential growth of L. monocytogenes in static and aerated batch culture with glucose as sole carbon source. The most suitable, judged on the basis of ease of preparation, growth rate and yield, was that of Trivett and Meyer (1971). This medium was shown to support growth of L. monocytogenes NCTC 7973 in continuous culture in a chemostat. A lytic phenomenon, noted with the same strain under anaerobic conditions and in batch culture in the chemostat, is discussed.     

Lipid composition of Staphylococcus aureus and its derived L-forms.

Two strains of Staphylococcus aureus (Newman and Tazaki) and their derived L-forms were cultured in serum-containing broth and the differences in their lipid compositions were analyzed. Cardiolipin accounted for more than 50% of the total phospholipid phosphorus in L-forms, but for less than 25% in parent bacteria. The cardiolipin content of L-forms was very high through all growth phases, although it increased gradually as growth proceeded. Significant amounts of cholesterol and its esters were present in parent strains and L-forms, all of which incorporated serum cholesterol into the cell membrane. On the other hand, they could be detected in the L-forms but not in the parent strains when they were cultured in serum-free broth. To examine the ability of L-forms to synthesize cholesterol, the cholesterol content of L-forms cultured in serum-free broth was compared with that of the medium. The results indicated that staphylococcal L-forms could synthesize cholesterol and its esters. These differences in lipid composition suggested that modification of membrane lipids may occur as an adaptational change in response to the disappearance of the cell wall.     

Production of lipase from Staphylococcus aureus grown on glucose media. II) Synthesis and release of enzyme in media containing glucose and cyclic AMP

Catabolite repression by glucose on lipase from Staphylococcus aureus is revealed as inhibition of the appearance of the enzyme in the surrounding medium but not as synthesis; dibutyryl cAMP results in an induction of cellular membrane permeation of the synthesized enzyme towards the external medium.     

Control of composition and activity of the photosynthetic apparatus of Rhodopseudomonas capsulata grown in ammonium-limited continuous culture

Rhodopseudomonas capsulata was grown either phototropically in the light or chemotrophically in the dark at oxygen tensions of 5 mm and 3 mm Hg in ammonium-limited continuous culture. During growth limitation bacteriochlorophyll content of cells and membranes varied dependent on growth rate drastically in chemotrophic cultures. Concomittantly, the ratio of membrane protein to total protein varied in the range of 30-41%. This dependence of membrane differentiation on growth rate was less evident in phototrophically grown cells. The incorporation of the bulk of bacteriochlorophyll was shown to be quantitatively correlated to the incorporation of 1-3 low molecular weight proteins with molecular weights in the range of 14 to less than 10 k daltons. Supported by similar findings of other authors it is proposed, that these proteins are to be attributed to the species of antenna bacteriochlorophyll and represent components of the photosynthetic apparatus. With decreasing growth rates the size of the photosynthetic unit with respect to the population of bacteriochlorophyll- and protein molecules was reduced subsequent to a reduction in the rate of incorporation of antenna-bacteriochlorophyll and the low molecular weight proteins, the reaction-center bacteriochlorophyll content of the membranes remaining constant. A parallel decrease in potential phosphorylating capacity was observed. It is concluded, that under these conditions, primary photochemical reactions in the reaction center were not the rate-limiting step in photophosphorylation. The interaction of growth limitation by an anabolic precursor (NH+4) and control of membrane differentiation by light intensity or oxygen tension is discussed.     

Comparison of selective media for coagulase-positive enterotoxigenic Staphylococcus aureus.

Different culture media were used to detect enterotoxigenic Staphylococcus aureus strains from peptone salt solution and inoculated minced meat. When recovery, reactions, and counting mistakes were considered, it was concluded that Baird-Parker agar was the best medium.     

Effect of specific growth limitations on cell wall composition of Staphylococcus aureus H

Conditions are described for the continuous culture of a derivative of Staphylococcus aureus H in a fully defined minimal medium in which cysteine is the sole amino acid. The effects of growth under various nutrient limitations on the composition and properties of the cell wall have been studied. The proportion of ribitol teichoic acid present in the wall, and the extent to which it is substituted with N-acetylglucosamine, varies in bacteria grown under different conditions as does the composition and extent of cross-linking of the peptidoglycan. Neither the derivative nor the original strain H produced teichuronic acid when grown under phosphate limitation.Non-Standard Abbreviation SDS Sodium dodecyl sulphate