Pathogenicity of stable L-phase variants of Staphylococcus aureus: failure to colonize experimental endocarditis in rabbits.
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Endocarditis was induced in the rabbit by the placement of a polyethylene catheter in the right heart. The catheter was filled with stable L-phase variants of Staphylococcus aureus to determine if the variant form would colonize the damaged endocardium and produce further tissue injury similar to that produced by the vegetative bacterial phase. No L-phase variants were recovered from cultures of blood or vegetations, although pure cultures of L-phase variants were obtained from all catheters, including one in place for 70 days. The vegetations were grossly similar in control animals with sterile media in the catheters and animals with catheters containing L-phase variants, although polymorphonuclear leukocytes and eosinophils were more frequently found in vegetations from animals inoculated with L-phase variants. Endocarditis was also induced in animals with vegetative S. aureus and then treated with penicillin G. Blood cultures in hypertonic media often grew vegetative S. aureus when there was no growth in routine media, but no L-phase variants were detected. Vegetative S. aureus, but not wall-defective variants, were isolated from vegetations of all treated animals. (+info)
Probable L-forms of Nocardia asteroides induced in cultured mouse peritoneal macrophages.
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Mouse peritoneal macrophages were infected with varying numbers of Nocardia asteroides 10905, and the fate of the ingested organisms was determined by viable plate count (VPC), light microscopy (LM), immunofluorescent microscopy (IM), and electron microscopy (EM). The results obtained with these methods differed. VPC indicated that intracellular Nocardia decreased in numbers during the first 12 days, followed by significant increases after 16 days. LM suggested that N. asteroides 10905 was slowly degraded by macrophages with no subsequent increases observed. In contrast, IM demonstrated large numbers of intracellular Nocardia throughout the experiment. EM studies of infected macrophages failed to demonstrate intact bacteria after 8 days; however, wall-less and spheroplast-like organisms were seen. These results suggested that N. asteroides 10905 was present within the macrophages in an altered form. By using hypertonic culture medium, we were able to isolate, from infected macrophages, organisms which exhibited many of the properties of bacterial L-forms. IM demonstrated these variants to be of nocardial origin. These altered forms also reverted to typical nocardial cells either spontaneously or upon transfer into broth. These findings indicate that N. asteroides 10905 is capable of existing within macrophages in an altered state. Further investigation is in progress to determine whether these altered forms represent L-forms or transitional-phase variants. (+info)
Detection of capsular antigen production in unencapsulated strains of Staphylococcus aureus.
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Of 91 compact-type strains of Staphylococcus aureus in regular serum soft agar (SSA), 82 converted diffuse-type growth in serum soft agar (pH adjusted to 6.0). With the addition of four different rabbit anticapsular sera (anti-type A, B, C, and D sera) in low pH (6.0) SSA, 21 strains of S. aureus showed compact-type colonial morphologies. Eleven, one, and one strains of S. aureus reacted singly with rabbit anticapsular sera types A, B, and C, respectively, and no strain reacted with rabbit anticapsular type D. Eight of the e strains reacted with both rabbit anticapsular sera types A and B. When the ability to absorb the converting activities of the antisera (changes of colonial morphologies of anticapsular sera in SSA) was quantitatively tested, 7- to 27-fold of these organisms were capable of absorbing the activities compared with the Smith diffuse organisms. These results suggest that even unencapsulated S. aureus strains are capable of producing capsular substance, although the capability is quantitatively different from strain to strain. (+info)
Incorporation of D-alanine into the membrane of Streptococcus pyogenes and its stabilized L-form.
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A principal aim of this study was to explain our earlier finding of a lack of d-alanine in the glycerol teichoic acid from the membrane of a stabilized L-form of Streptococcus pyogenes (B. M. Slabyj and C. Panos, 1973. J. Bacteriol. 114:934-942). It was found that the incorporation of d-alanine into the membrane teichoic acid of S. pyogenes requires either supernatant fraction or two enzymes from supernatant fraction, stimulator (d-alanine activating enzyme) and d-alanine:membrane acceptor ligase, plus membrane fragments, ATP and Mg(2+). A similar system from the L-form is inoperative. Also, no incorporation is observed with L-form or coccal supernatant fractions when L-form membranes are used. However, d-alanine incorporation is observed when L-form enzymes are used with membrane fragments from the parental streptococcus. Thus, the L-form possesses the required soluble components for d-alanine incorporation but the L-form membrane cannot function as acceptor even though it contains d-alanine-deficient membrane teichoic acid. These results suggest that a defect has occurred in the membrane of this stabilized L-form for d-alanine incorporation into membrane teichoic acid. (+info)
Osmotic stability and sodium and potassium content of L-forms of Streptococcus faecalis.
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A stable L-form of Streptococcus faecalis (T(53)) was transferred in media containing decreasing concentrations of sucrose until it grew in medium without added osmotic stabilizer. This L-form (designated T(53I)) was compared with T(53) from which it was derived. The survival of these two L-forms suspended at different osmolalities showed that maximal survival for T(53I) was 350 to 400 milliosmolal and for T(53) was 900 to 1,000 milliosmolal. Both peaks were at the osmolality of their growth media. Measurement of intracellular potassium and sodium showed that the concentration of these ions was reduced in T(53I). (+info)
Phagocytosis of microbial L-forms.
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Little data are available regarding L-form phagocyte interactions. Due to difficulty with more conventional methods, a serum-free technique of surface phagocytosis employing cellulose ester micropore filters, fluorochrome staining, and reflecting fluorescence microscopy was developed. Intracellular localization of L-frms was confirmed by electron microscopy. In timed experiments, phagocytosis of a group D Streptococcus (ATCC 23241) and its L-form (T53I) were maximal at 30 min, although the bacteria were consistently better phagocyted. Phagocytosis of an Escherichia coli (strain Yale) and its L-form was also demonstrated with the same technique. The lesser phagocytosis of L-forms may be related to their lower chemotactic activity. (+info)
Characterization of a stable L-form of Bacillus subtilis 168.
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A stable L-form of Bacillus subtilis 168 (sal-1) has been isolated which grows and divides logarithmically in liquid medium with a generation time of 60 min. This mutant does not synthesize cell wall as evidenced by chemical, biochemical, and morphological analyses. Antibiotics which specifically inhibit cell wall biosynthesis do not affect the growth of the L-form. Significant differences exist between the membrane proteins of the bacillary form and the L-form. The relative profile of membrane proteins varies with the salt concentration of the medium in both the L-form and the bacillary form. (+info)
Surface structure of intact cells and spheroplasts of pseudomonas aeruginosa.
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This report describes the ultrastructural features of Pseudomonas aeruginosa after freeze-etching of intact cells and enzymatically prepared spheroplasts. Freeze-etching of intact cells revealed two convex layers of the cell wall and particles within the hydrophobic interior of the cell membrane. Areas of the membrane free of particles were sometimes elevated in the form of rather large dome-shaped structures. Spheroplasts were formed from intact cells by the addition of trypsin to a reaction mixture of lysozyme and ethylenediaminetetraacetic acid. Spheroplasts contained the outer lipoid layer of the cell wall. It was possible to observe this cell wall layer in freeze-etch preparations of spheroplasts. The spheroplast membrane like that of intact cells was cleaved along a central plane to expose particles and particle-free areas. (+info)