Search

The distribution of polysaccharide-forming activity from sucrose was investigated in cultures of three strains of Streptococcus mutans by using an assay which conveniently determines total polysaccharide. The enzymatic activity for polysaccharide formation from sucrose is almost exclusively extracellular. The ratio of the fructan to glucan in the polysaccharide differs among the three strains investigated. The enzymatic activity for the formation of polysaccharide from sucrose has been shown to be bound to the cell-free polymer itself.

This paper describes the properties and use of a fiber optic probe as an attachment to a spectrophotometer and its use for measurements in solutions and turbid suspensions. Measurements of a standard were identical when a spectrophotometer equipped with the probe was used or when a spectrophotometer was used in a conventional manner. The probe was calibrated for turbidimetric measurements with microorganisms by relating the apparent absorbancy measured on the spectrophotometer to the dry weight of each species of organism. Continuous measurements were made of the turbidity of growing cultures of Escherichia coli, Streptococcus mutans, and Saccharomyces cerevisiae. Transient changes in cell mass were observed in some cultures during continuous monitoring of growth. The data were recovered in a manner which allowed direct computer processing.

Rogosa, M., (National Institutes of Health, Bethesda, Md.), M. I. Krichevsky, and F. S. Bishop. Truncated glycolytic system in Veillonella. J. Bacteriol. 90:164-171. 1965.-Intact Veillonella cells do not utilize carbohydrates for growth, nor are carbohydrates fermented. In cell extracts, there is no detectable glucokinase or fructokinase. Cell extracts do not degrade glucose or fructose unless supplemented with yeast hexokinase. Under these conditions, triose phosphates are formed in the presence of a hydrazine trap. When glucose-C(14) plus added hexokinase or fructose-1,6-diphosphate-C(14) was incubated with cell extracts, the production of CO(2), acetate, pyruvate, propionate, and lactate was detected. It is concluded that, except for a hexokinase, all the activities required for a glycolytic system are present.

Cultures of the amoeboid slime mold, Dictyostelium discoideum, were grown in the presence of various antibiotics and antimetabolites. Results indicate that many antibiotics may be used in amoeba cultures to help minimize contamination (for example: polymyxin B, kanamycin, tetracycline, neomycin, oxacillin, ampicillin, cephalothin, and erythromycin). A few potent antibiotics were found to be unsuitable (colistin, nystatin, amphotericin B, actinomycin, and cycloheximide); the mechanisms of the inhibitions observed were not investigated further.

A simple, sensitive, and rapid method is presented for the determination of deoxyribonucleic acid (DNA) in both gram-positive and gram-negative bacteria. It is based upon the fluorometric determination of DNA with ethidium bromide after alkaline digestion of the bacteria to hydrolyze the interfering ribonucleic acid. The assay takes less than 2 hr. Its sensitivity is at least 0.2 mug of DNA in a final solution of 4 ml and it uses commonly available filter or double monochromator fluorometers. Judicious choice of light source and filters allows an additional 10-fold increase in sensitivity with a filter fluorometer. Turbidity caused by bacteria or insoluble polysaccharides does not interfere with the fluorescence measurements. There was no significant difference between the results obtained with this method and those obtained with the indole and diphenylamine methods when these assays were applied to Escherichia coli and sucrose- or glucose-grown Streptococcus mutans. The method was also tested by determining the specific growth rate of E. coli. This new procedure should be especially useful for the determination of bacterial DNA in dilute suspensions and for the estimation of bacterial growth or DNA replication where more conventional methods are not applicable or sensitive enough.

A specific antibody directed against adenosine 3':5'-cyclic monophosphate diesterase (c-AMP diesterase; EC 3.1.4.c), which is secreted by the amoeboid slime mold, Dictyostelium discoideum, was prepared in rabbits. Purified gammaglobulin fractions that contain the antibodies inhibited the activity of cAMP diesterase preparations in vitro and interfered with aggregation and development in vivo. While cells that were treated with antibody were unable to aggregate because of the inability to destroy cAMP, they aggregated normally when washed free of antibody.

3',5'-Cyclic nucleotides are acrasins for the cellular slime mold, Dictyostelium discoideum; they have chemotactic activity on the myxamoebae at very low concentrations. However, not all cyclic nucleotides can evoke the adhesiveness necessary for aggregate formation. 2',3'-Cyclic nucleotides and dibutyrylcyclic adenosine monophosphate are not acrasins, though they enhance the rate of differentiation and morphogenesis. The addition of 3',5'-cyclic nucleotides to agar test plates stimulates the rate of morphogenesis. While the stimulatory effect of cyclic nucleotides is concentration-dependent, it is fairly uniform between pH 5 and 7, with an optimum at pH 6. The acrasins may be metabolized by extracellular phosphodiesterase to 5'-nucelotides which may then stimulate differentiation and morphogenesis.