Effect of Streptococcus pyogenes on Tissue Cells

Human tissue cell lines from each of the three primary germinal sources, ectoderm (conjunctiva and carcinoma of the buccal mucosa), entoderm (intestine and liver), and mesoderm (heart and monocytes) were inoculated with group A Streptococcus pyogenes, Staphylococcus aureus, and group D streptococci and were then observed. In addition, the effect of these bacteria on mouse fibroblasts was studied. All of the cell lines appeared to be equally susceptible to damage, but damage to the cells by S. pyogenes occurred only when living, actively multiplying bacteria were in contact with the tissue cells. Streptococcal products in the form of "used" growth medium had no observable effect on the cells. Cytopathogenic effects were first noticed about the time one would expect the bacteria to have reached the end of the log phase of growth. No damage to the tissue cells was noted when group A streptococci were separated from the cells by membrane filter diffusion chambers or dialyzing membranes, but a membrane did not protect cells from deleterious effects of staphylococci or group D streptococci. Group A streptococci survived in the tissue culture medium, but multiplication did not occur unless living tissue cells were present.     

Formation of Protoporphyrin from Coproporphyrinogen in Extracts of Various Bacteria

The enzyme system capable of converting coproporphyrinogen to protoporphyrin was demonstrated in the soluble fraction of extracts of Pseudomonas fluorescens grown aerobically, of P. denitrificans grown anaerobically under denitrifying conditions, and of Escherichia coli grown both aerobically and anaerobically. Protoporphyrin accumulation by each of these extracts occurred only if the assay was conducted aerobically. Attempts to replace this oxygen requirement with several alternate electron acceptors were not successful. The conversion of coproporphyrinogen to protoporphyrin could not be demonstrated in extracts of the heme-containing organisms Staphylococcus epidermidis and Bacillus subtilis.     

Effect of antibiotics on ribosomal RNA and proteins from Streptococcus pyogenes

Summary Cells of Streptococcus pyogenes were prepared under rigid conditions. The microorganisms were then incubated for 3 hours in the presence or absence of chloramphenicol, actinomycin or puromycin. RNA, ribosomal fraction and ribosomal proteins were isolated from the cells. The materials were invesigated with the help of infra red spectroscopy using the potassium bromide pellet method. Quantitative differences in the 1750–1500 cm^-1 region were observed with materials treated with the antibiotics. Synthetic mixtures of ribosomal RNA with progressively larger amounts of ribosomal proteins show analogous changes, namely a progressive increase in the strength of the 1650 cm^-1 band relative to the 1685 cm^-1 band, and an increase in the 1535 cm^-1 band. The analytical results obtained with the ribosomal RNA isolated from S. pyogenes treated with antibiotics indicated increased amounts of proteins which could not be removed by the applied extraction method. The evidence presented suggests a change in the binding between ribosomal RNA and ribosomal proteins in the material isolated from the antibiotic treated microorganisms. The I. R. spectroscopy seems to be an useful tool in the investigation of some aspects of biological materials.     

Effect of pH on In Vitro Phagocytosis of Streptococcus pyogenes

Phagocytosis experiments were performed with mouse peritoneal leucocytes (MPL). The natural pH of the mouse peritoneal cavity was found to be between 6.1 and 6.3. The phagocytic and intracellular killing activities of MPL by pH variations was studied. It was observed that the optimal ingestion and intracellular killing of bacteria is at the natural pH of the peritoneal cavity.     

Effect of Incubation Temperature on T-Agglutination Typing of Streptococcus pyogenes

The temperature of incubation affected the typability of beta-hemolytic group A streptococci by T-agglutination tests. When strains could not be typed after routine incubation at 30 C, they were incubated at 22 to 25 C, and nearly a 10% increase in typability was achieved. The clinical source of the strains was related to their typability. Incubation at the lower temperature was required for successful typing of higher percentages of strains from the skin and other clinical sources than from the throat. Sixty per cent of the skin strains were represented by six serotypes. Of these, 53% of the strains required incubation at 22 to 25 C before they could be typed.     

Effect of date extract on growth and hemolytic activity of Streptococcus pyogenes

The effect of date extract on growth and hemolytic activity of S. pyogenes was examined. It was found that 5% DE caused 78 % growth inhibition. However, 20% DE inhibited the growth by 86%. 5% DE inhibited hemolysin and streptolysin O activities by 43% and 24% respectively,while 20% caused 95 and 91 %inhibition.     

Effect of Gramicidin on Methanogenesis by Various Methanogenic Bacteria

Methanogenesis by Methanobacterium thermoautotrophicum strains was extremely sensitive to gramicidin, total inhibition being observed at 0.2 μg/ml. In contrast, methane synthesis by Methanococcus voltae, Methanogenium marisnigri, Methanosarcina mazei, and Methanospirillum hungatei were resistant to the highest concentrations of gramicidin tested (40 μg/ml), although spheroplasts of Methanospirillum hungatei were extremely sensitive. Other species tested showed intermediate sensitivity to gramicidin, methanogenesis inhibition occurring at 4 to 20 μg/ml.     

The NAD-glycohydrolase (nga) gene of Streptococcus pyogenes

The gene for NAD-glycohydrolase (nga) of group A streptococci (Streptococcus pyogenes) was identified and shown to be located immediately adjacent to the gene for streptolysin O (slo). The nga gene contains 1341 base pairs and encodes a protein of 447 amino acids, including an N-terminal signal peptide. Results from analysis with the polymerase chain reaction indicated that the nga gene is present in all of the strains tested. Functional extracellular NAD-glycohydrolase, also known as NADase, was detected among a wide variety of clinical isolates and known laboratory strains and shown to be present in 72% of 100 strains examined. In contrast, 92% of strains isolated from patients with invasive streptococcal infections were positive for NADase production.     

The effect of the flavonol morin on adhesion and aggregation of Streptococcus pyogenes

The effect of the flavonol morin on Streptococcus pyogenes biofilm growth was determined using a static biofilm model, in which reduced biofilm biomass was observed in the presence of morin, suggesting that morin inhibited biofilm development. Morin at concentrations exceeding 225 μM had the greatest impact on biofilm biomass causing reductions of up to 65%, which was found to be statistically significant. Morin was also shown to induce rapid bacterial aggregation. Approximately 55% of S. pyogenes in liquid suspension aggregated when incubated with morin at concentrations of 275 and 300 μM for 120 min, compared to the control group in which only 10% of the cells aggregated, this was also shown to be statistically significant.     

Fluoride modifies adhesion of Streptococcus pyogenes

Streptococcus pyogenes grown in the presence of subinhibitory concentrations of sodium fluoride had a diminished ability, compared to control cells, to adhere to buccal cells, collagen, fibronectin, and laminin. In addition, sodium fluoride was a competitive inhibitor of streptococcal adhesion to collagen and fibronectin, but not laminin. It is suggested that sodium fluoride may be useful in therapy or prophylaxis in infections involving group A streptococci.