Biomedical subjects
Publications and source records attributed to J Mikucki.
In eleven strains of genus Enterococcus the growth kinetics and hydroxamate siderophore production in iron-deficient medium were investigated. The classical pattern of bacterial growth kinetics and siderophore synthesis was observed.
In eleven strains of genus Enterococcus relation between hydroxamate siderophore production and growth in iron-restricted liquid medium was investigated. Fe(III) deficiency caused moderate decrease of growth rate: the exponential growth phase was prolonged and yield of bacterial mass was lower. The hydroxamate siderophore was produced by all strains but occurred at different phase of growth. Only part of strains showed classical pattern of growth kinetic and siderophore synthesis.
Under iron restricted conditions enterococcal strains could utilize in vitro several animal body iron sources in form of bovine hemoglobin, hemin, lactoferrin and transferrin, ovotransferrin, horse myoglobin, ferritin and cytochrome C. Spectrum of utilized iron sources was not depended on species affiliation and origin of strains.
Among 20 strains of genus Enterococcus isolated from different sources 12 strains multiplied in iron unsupplemented normal human serum. The excess of iron did not increase the rate of multiplication. Three strains survived in serum and growth of five strains was inhibited. The siderophores production had no effect on the multiplication of enterococci in serum unsupplemented with iron.
The study was undertaken to investigate the ability of enterococci to assimilate iron via siderophores of bacteria living in the same habitats in the human organism. The potential recipients of exogenous siderophores were six Enterococcus faecalis and six Enterococcus faecium strains, isolated from clinical materials of human origin. The donors of siderophores were Gram-negative rods (various species of the Enterobacteriaceae, Pseudomonas and Acinetobacter) and Gram-positive cocci (various species of Staphylococcus and Streptococcus). All of the investigated E. faecium and only two E. faecalis strains demonstrated the ability to utilize the siderophores of the aforementioned bacterial groups, predominantly the chelators of Gram-negative rods, those of Gram-positive cocci were utilized to a smaller extent. Four recipient strains from E. faecalis species did not demonstrate the ability to utilize siderophores synthesized by all of 40 investigated donor strains.
The influence of Cefodizime (CDZ) on in vitro activity of polymorphonuclear leukocytes (PMNL) from healthy subjects was assessed. Preincubation with CDZ enhanced phagocytosis and intracellular killing of Staphylococcus aureus by PMNL. Contrary to numerous clinical reports, no significant effect of CDZ preincubation on PMNL response to n-formyl-methionyl-leucyl-phenylalanine was found with respect to intracellular calcium changes, degranulation, hydrogen peroxide production, and chemiluminescence. These results suggest that augmented microbicidal activity of PMNL is not related to the enhanced production of reactive oxygen species in healthy subjects.
In 70 strains of the genus Enterococcus belonging to 16 species and isolated from clinical material, animals and the environment, a hydroxamic class of siderophores was detected with the aid of chemical and biological tests. A correlation between siderophore activity and species affiliation was found: E. faecalis strains showed the highest siderophore activity. A correlation between siderophore activity and pathogenicity could not be confirmed.
A total of 237 strains of staphylococci belonging to 27 species were tested for susceptibility to natural and synthetic iron-chelators. Coagulase-negative staphylococci were much more susceptible than coagulase-positive ones: 82 out of 148 coagulase-negative strains were susceptible to desferrioxamine B, rhodotorulic acid and ovotransferrin and 7 out of 89 coagulase-positive strains. A correlation between the susceptibility to iron-chelators species affiliation and the origin of strain was not found.
The functional interchangeability of siderophores was tested among 62 strais belonging to 12 species of genus Enterococcus. Most investigated strains were from E. faecalis and E. faecium species. The majority of examined enterococcal strains appeared highly resistant to EDDA (ethylene-di-amine-di-ortho-hydroxyphenylacetic acid), therefore the group of sensitive strains involved only 11 used as indicator strains. The determination of interchangeability of siderophores within enterococcal strains was performed using EDDA-agar media into which the indicator strains were included. Test colonies (donor strains) were applied to the surface of the media to determine whether the indicator organisms could obtain the required iron for growth by utilizing chelators from the test colony. Only two strains: E. solitarius DSM 5634 and E. pseudoavium DSM 5632 did not demonstrate the ability to utilize siderophores synthesized by all investigated strains. The other tested indicator strains appeared to be recipients of siderophores from 20-52 donor enterococcal strains. The ability to exchange siderophores in enterococci was found as the feature characterizing individual strains.
The ability of iron utilizing by means of staphylococcal siderophores by bacteria belonging to genera: Acinetobacter, Corynebacterium, Curtobacterium, Clavibacter, Bacillus and Mycobacterium was investigated. The staphylococcal donor strains (18 species) used in these experiments were characterized by the ability to utilize siderophores produced by various strains belonging to aforenamed genera. The utilization of staphylococcal siderophores was studied on agar media in which minimally effective concentrations of ethylenediaminedi-ortho-hydroxyphenylacetic acid (EDDA) were used to inhibit indicator strains. Test colonies (staphylococcal) were applied to the surface of the media to determine whether the indicator organisms could obtain the required iron for growth by utilizing chelators from the test colony. The growth inhibition by EDDA of most strains from the Acinetobacter rods and from the coryneform-organisms (plant pathogen) genera, and strains from the species: B. subtilis, M. phlei, M. smegmatis, M. fortuitum was reversed by staphylococcal siderophores. None of the staphylococcal strains investigated, had the ability to exchange siderophores with strains from the species: C. pseudodiphtheriticum, Corynebacterium ANF group, B. megaterium, M. vaccae, M. chitae and M. parafortuitum.
In 70 enterococcal strains of diverse origin belonging to 16 species a hydrooxamate class siderophore was detected with chemical and biological tests. A correlation between hydrooxamate siderophore production, species affilIation and pathogenicity of enterococci was not found.
The ability of iron utilizing by means of siderophores (extracellular exochelins and cell-associated mycobactins) produced by mycobacteria (7 stains) by 24 staphylococcal strains was investigated. The mycobacterial donor strains belonged to rapid growing species: M. fortuitum, M. smegmatis, M. aurum, M. vaccae, M. chitae, M. phlei, M. parafortuitum. The utilization of mycobacterial siderophores was studied on agar or liquid media in which minimally effective concentrations of ethylene diamine di-ortho-hydroxyphenyl acetic acid (EDDA) were used to inhibit indicator staphylococcal strains. Mycobacterial siderophores (exochelins or mycobactins) reversed the inhibition of the majority (22/24) staphylococcal strains. Most of strains utilized exochelins from M. phlei as well as mycobactins from M. parafortuitum and M. chitae.
In previous study we demonstrated that Staphylococci aureus clinical and environmental isolates differ by siderophore production (Lisiecki et al., 1997), the aim of the present study was to check a possible role of siderophore-dependent iron acquisition system of outcome of staphylococcal diseases. The systemic and local staphylococcal infections were induced in mice by inoculation of three S. aureus strains differing by siderophore production. We found that S. aureus B 47 strain characterized by enhanced siderophore activity was more virulent in both systemic and local infection models and it was more resistant to anti-bacterial activity of neutrophils than S. aureus B 63 and B 32 strains expressing weaker siderophore production. The results suggest that effective siderophore-dependent iron acquisition system may be beneficial to S. aureus strains in their pathogenic activity in vivo.
The ability of iron utilizing by means of siderophores produced by donor strains--the members of the genus Acinetobacter (8 strains) by 24 staphylococcal strains was investigated. All the donor strains synthesized hydroxamate class siderophores and six strains also catecholate class. The majority of staphylococcal strains could utilize these siderophores. Most strains utilized siderophores from A. juni 321 and A. johnsonii 349 strains. Only three staphylococcal strains were not be able to utilize siderophores from all donor strains.
Under iron-restricted conditions staphylococcal strains could utilize in vitro several animals body iron sources in form of bovine haemoglobin, hemin, lactoferrin and transferrin, ovotransferrin, horse myoglobin ferritin and cytochrome C. Spectrum of utilized iron sources was not dependent on species affiliation and kind of siderophores system. Strains isolated from clinical materials utilized largest spectrum of animal iron body sources.
The aim of this study was to investigate the influence of iron present in the growth medium of Staphylococcus aureus on the bacterial adhesion to collagen. The experiments were extended to determinate the siderophore production and to examine the S. aureus isolates surface hydrophobicity. The addition of iron to metal deficient defined medium causes the change in hydrophobicity of the examined S. aureus strains surfaces from hydrophilic to hydrophobic. The presence of iron in staphylococcal growth medium alters also the adhesion to the surface covered with collagen. Four out of six S. aureus strains adhere to collagen weaker when cells come from iron-rich medium. Majority of tested strains produce markedly less of siderophores in media containing the excess of iron (1 and 10 microM Fe) and there is no staphylococcal siderophore activity in the growth medium with a very high concentration of this compound (120 microM Fe). The obtained results indicate that the iron-stressed conditions influence the staphylococcal adhesion to collagen.
The ability of iron utilizing by means of siderophores produced by donor strains-Coryne-bacterium and coryneform organisms (8 strains) by 24 staphylococcal strains was investigated. All the donor strains synthesized catecholate class siderophores and two strains also hydroxamate class. The majority of staphylococcal strain could utilize these siderophores. Most of strains utilized siderophores from Corynebacterium pseudodiphtheriticum and Corynebacterium aquaticum as well as chelators from plant pathogens-coryneform organisms. Only two staphylococcal strains were not be able to utilize siderophores from all donor strains.
Under iron-restricted conditions staphylococcal strains could utilize in vitro human body iron sources in form of haemoglobin, haemin, transferrin and lactoferrin. Iron was taken up after proteolytic degradation of the binding protein and by means of siderophores. Siderophores were identified as hydroxamates.