Phagocytosis of chloramine B sensitive and resistant staphylococci isolated from healthy persons and patients

Phagocytic reaction with respect to antibiotic and chloramine B sensitive and resistant staphylococci isolated from healthy persons and patients, air and stock of medical institutions was studied on albino mice. It was shown that the staphylococcal isolates included strains simultaneously sensitive to antibiotics and chloramine, sensitive to antibiotics and resistant to chloramine, resistant to antibiotics and sensitive to chloramine and simultaneously resistant to antibiotics and chloramine. Activity, intensity and completeness of phagocytosis by leucocytes from mouse abdominal cavity exudates with respect to the staphylococcal strains sensitive to antibiotics and resistant to chloramine, resistant to antibiotics and sensitive to chloramine and simultaneously resistant to antibiotics and chloramine were lower than the values of the phagocytic reaction with respect to the isolates simultaneously sensitive to antibiotics and chloramine. This suggested that not only antibiotic resistance of microbes but also their resistance to disinfectants could be referred to complicating factors of hospital infections.     

Antibiotic Therapy of Staphylococcal Infections

The antibiotic treatment of staphylococcal infections remains a problem. Isolation of the organism and sensitivity testing are necessary in the choice of antibiotic. Penicillin G is the most effective penicillin against non-penicillinase-producing staphy-lococci; for the penicillinase producers there is very little to choose between the semisynthetic penicillins, methicillin, cloxacillin, nafcillin and oxacillin. For patients who are hypersensitive to penicillin, the bacteriostatic drugs (erythromycin, novobiocin, tetracycline, chloramphenicol, oleandomycin) are useful for mild infections, while for more severe illness the bactericidal drugs (vancomycin, ristocetin, kanamycin, bacitracin, neomycin) have been used successfully. Acute staphylococcal enterocolitis is probably best treated by a semisynthetic penicillin. Other antibiotics which have been found useful, with clinical trials, for staphylococcal infections are cephalosporin, fucidin, cephaloridine and lincomycin. The latter drug has been reported of value in the treatment of osteomyelitis. There is little justification for the prophylactic use of antibiotics to prevent staphylococcal infection. Surgical drainage is still an important adjunct in the treatment of many staphylococcal infections.     

Methods for detecting drug-resistant staphylococci in the human body under natural conditions]

Two methods were compared for determination of drug resistant staphylococci on the nasal mucosa of patients, i. e. the routine method for determination of staphylococcal sensitivity to antibiotics and the method of direct plating out of the starting material onto agarized media containing antibiotics. Staphylococci resistant to streptomycin, tetracycline, chloramphenicol, erythromycin, monomycin, axacillin and less frequent to penicillin were found more often with the 2nd method. A method of proportions was developed for testing sensitivity of staphylococci in purulent inflammatory foci of the patients. It provided characterization of the staphylococcal population from the foci by the number of the antibiotic resistant microbial cells.     

Characterization of pACK4, a mobilizable plasmid from Staphylococcus simulans biovar staphylolyticus

Staphylococcus simulans biovar staphylolyticus, the lysostaphin-producing organism, contains five plasmids designated pACK1–pACK5. pACK4 was found to be relaxable and to share sequence similarity with a number of well-characterized mobilizable plasmids from other staphylococci. All mobilizable staphylococcal plasmids characterized to date mediate resistance to various antibiotics, but pACK4 is unique because it contains no recognizable antibiotic resistance genes. pACK4 was found to contain an origin of transfer (oriT) region that shares inverted repeat regions and the same nic site as several other mobilizable staphylococcal plasmids. The presence of this conserved oriT region suggested that pACK4 might be mobilized in the presence of a conjugative plasmid. Filter mating studies revealed that pACK4 was mobilized by the conjugative plasmid pGO1. In addition, pACK4 was found to be virtually identical to the recently described plasmid pVGA from Staphylococcus aureus, except that pVGA contains an additional region (vgaA) that confers resistance to pleuromutilin, streptogramin A, and lincosamide. The high sequence similarity among pACK4, pVGA, and several previously described mobilizable staphylococcal plasmids suggests a common origin for these plasmids.     

Synergism between antibiotics and plant extracts or essential oils with efflux pump inhibitory activity in coping with multidrug-resistant staphylococci

The alarming growth of the number of antibiotic resistant bacteria and in the same time limited possibilities to develop new antimicrobial compounds, lead to an urgent need to keep the sensitivity of bacteria against currently used antibiotics. Bacterial efflux pumps are an important mechanism of antibiotic resistance as the bacteria use efflux pumps for the extrusion of different types of antibiotics and chemicals. The knowledge about inhibitors of efflux pumps from natural sources suggests that this mechanism may be a good target for new drugs based on synergistic interactions of antibiotics with plant extracts, essential oils, or their constituents with efflux pump inhibitory activity. This review summarizes the current knowledge of staphylococcal efflux pumps and potential strategies to overcome them. Natural inhibitors of efflux pumps and their synergistic interactions with antibiotics are summarized.     

An examination of the inhibitory effects of three antibiotics in combination on ribosome biosynthesis in Staphylococcus aureus

Although a number of different antibiotics are used to combat staphylococcal infections, resistance has continued to develop. The use of rifampicin and ciprofloxacin in combination with azithromycin, known for its inhibitory effects on the bacterial ribosome, can create potential synergistic effects on ribosomal subunit synthesis rates. In this work, combination antibiotic treatments gave a significant decrease in cell numbers following growth in the presence of ciprofloxacin or rifampicin with azithromycin compared to those grown with azithromycin or rifampicin alone. DNA, RNA and protein synthesis rates were reduced with single antibiotic treatments and showed further decreases when drug combinations were used. 70S ribosome levels were reduced with every antibiotic treatment. DNA gyrase subunits A and B showed significant decreases for double and triple antibiotic-treated samples. Ribosomal subunit synthesis rates were diminished for each different antibiotic combination. Turnover of 16S and 23S rRNA was also observed in each case and was stimulated by antibiotic combinations. The frequency of spontaneous resistance was reduced in all double selections, and no triply resistant mutants were found.     

Study of beta-lactamase activity of staphylococci from different sources

The beta-lactamase activity of staphylococci isolated from the nasopharynx and skin of children with destructive affections of the lungs and from blood of patients with cardiovascular diseases subjected to surgical operations was determined with acidometric and microbiological procedures. Interrelation between synthesis of beta-lactamase by the staphylococcal strains and their resistance to beta-lactam antibiotics was demonstrated. No correlation of the antibiotic resistance and the taxonomic position of the staphylococcal strains was observed.     

Susceptibility of staphylococcal biofilms to enzymatic treatments depends on their chemical composition

Bacterial infections are serious complications after orthopaedic implant surgery. Staphylococci, with Staphylococcus epidermidis as a leading species, are the prevalent and most important species involved in orthopaedic implant-related infections. The biofilm mode of growth of these bacteria on an implant surface protects the organisms from the host’s immune system and from antibiotic therapy. Therapeutic agents that disintegrate the biofilm matrix would release planktonic cells into the environment and therefore allow antibiotics to eliminate the bacteria. An addition of a biofilm-degrading agent to a solution used for washing–draining procedures of infected orthopaedic implants would greatly improve the efficiency of the procedure and thus help to avoid the removal of the implant. We have previously shown that the extracellular staphylococcal matrix consists of a poly-N-acetylglucosamine (PNAG), extracellular teichoic acids (TAs) and protein components. In this study, we accessed the sensitivity of pre-formed biofilms of five clinical staphylococcal strains associated with orthopaedic prosthesis infections and with known compositions of the biofilm matrix to periodate, Pectinex Ultra SP, proteinase K, trypsin, pancreatin and dispersin B, an enzyme with a PNAG-hydrolysing activity. We also tested the effect of these agents on the purified carbohydrate components of staphylococcal biofilms, PNAG and TA. We found that the enzymatic detachment of staphylococcal biofilms depends on the nature of their constituents and varies between the clinical isolates. We suggest that a treatment with dispersin B followed by a protease (proteinase K or trypsin) could be capable to eradicate biofilms of a variety of staphylococcal strains on inert surfaces.     

In vitro testing of gentamicin-vancomycin loaded bone cement to prevent prosthetic joint infection

Sepsis is a greatly feared complication of total joint arthroplasty. One key question is how to prevent perioperative bacterial adherence, and therefore the potential for infectious complications. The objective of our study was to appraise the emerging capacity of staphylococcal survival on prosthetic materials and to analyze the in vitro effects of gentamicin and vancomycin loaded polymethylmethacrylate (PMMA) cement on bacterial adherence and growth. Hospital acquired staphylococcal strains were systematically inoculated on four orthopedic materials (ultrahigh molecular weight polyethylene, PMMA without antibiotic, commercially produced PMMA loaded with gentamicin, and manually mixed PMMA loaded with gentamicin and vancomycin). Staphylococci were identified using culture and biochemical tests. The inoculated material was allowed to incubate in a liquid broth growth media and subsequently prepared for scanning electron microscopy and bacterial growth quantification. Materials without antibiotics showed evidence of staphylococcal growth. PMMA loaded with only gentamicin grew methicillin-resistant Staphylococcus aureus. Gentamicin-vancomycin loaded PMMA completely inhibited any bacterial growth. Low-dose gentamicin-vancomycin loaded PMMA prevents staphylococcal colonization better than commercially manufactured PMMA loaded with gentamicin. We recommend this combination in high-risk procedures and revision surgeries requiring bone cement.     

Action of heliomycin on the incorporation of labelled protein precursors and nucleic acid in Staphylococcus aureus cells]

In concentrations of 0.05-0.10 gamma/ml geliomycin suppresses inclusion of C14-uracyl into the acid-soluble fractions of the staphylococcal cells. Inclusion of C14-leucine at the same concentrations of the antibiotics was suppressed to a lesser extent. The above concentrations of geliomycin had practically no effect on inclusion of C14-thimine. Suppression of C14-uracyl inclusion was observed in 5 minutes of the cell incubation with the antibiotic, a significant decrease in C14-leucine inclusion being observed only in 30 minutes of the incubation. The effect of geliomycin on the staphylococcal cells within the concentrations inhibiting the microbial growth and suppressing inclusion of labeled uracyl and leucine into the cells was of the bacteriostatic nature. It was supposed that the antibacterial effect of geliomycin was based on suppression of RNA synthesis.     

Biological properties of clinical staphylococcal strains with a varying number of antibiotic resistance markers]

The study of a number of biological properties of 1881 clinical strains of Staphylococcus showed that in the group of the antibiotic resistant staphylococci there was a tendency for different manifestation of some biological properties depending on the number of the resistance determinants. The staphylococcal strains resistant to 5--7 antibiotics differed from those resistant to a less number of the drugs by greater manifestation of the pathogenicity properties: lecithinase, hyaluronidase and hemolytic activity.     

Characteristics of hospital strains of Staphylococcus]

Some properties of hospital staphylococcal strains isolated from the patients with purulent postnatal mastitis were studied. It was found that all the strains of the phage type 80.81 widely distributed in the obstetric hospitals were polylysogenic and polyresistant to antibiotics. The resistance markers in most of the isolates were located on the plasmids. Elimination of the plasmids carrying the antibiotics resistance markers resulted in a simultaneous change in the strain lysogenic spectrum while the phage type and fermentative properties, such as production of plasmocoagulase, DNAase and lecithinase did not change. The capacity for hemotoxin production was lost simultaneously with antibiotic resistance in 1 out of 17 cultures tested.     

Bile acids in the fetal rat: Effect of maternal bile duct ligation

The effect of bile duct ligation during pregnancy in rats (thereby increasing maternal plasma bile acids levels) on the bile acid content and composition in the fetus was examined. In spite of 30-fold increase in maternal plasma cholic acid, the bile acid content in the fetus of bile duct ligated rats was significantly lower (P <0.05) with a significant reduction in cholic acid content. Plasma cholesterol levels of fetuses from bile duct ligated rats were also significantly lower (p <0.05). In addition to the commonly expected bile acids, gas-liquid Chromatographic analysis of the fetal bile acid pool showed peaks corresponding to several secondary bile acids. These results suggest that the transfer of primary bile acids of maternal origin into the fetus is minimal.     

Effect of antibiotics on the formation of specific antibodies.

Immune response to staphylococcal haemolysin and Haemophilus influenzae capsular antigen administered simultaneously with antibiotics was studied in rabbits. In addition to specific humoral antibodies, the quantitative values of IgG, IgA, IgM and C'3 complement were determined. Statistically significant deficiency of immune response was observed in all cases in animals which were given the antigen with the antibiotic in comparison with the controls which were immunized by the antigen alone. Statistically significant differences were also observed in the levels of immunoglobulins (mainly IgG and IgM) in animals which were given antigens simultaneously with antibiotics in comparison with animals which were given antibiotics alone.     

The AcrAB RND efflux system from the live vaccine strain of Francisella tularensis is a multiple drug efflux system that is required for virulence in mice

The ability of bacterial pathogens to infect and cause disease is dependent upon their ability to resist antimicrobial components produced by their host, such as bile acids, fatty acids and other detergent-like molecules, and products of the innate immune system (e.g. cationic antimicrobial peptides). Bacterial resistance to the antimicrobial effects of such compounds is often mediated by active efflux systems belonging to the resistance-nodulation-division (RND) family of transporters. RND efflux systems have been implicated in antibiotic resistance and virulence extending their clinical relevance. In this report the hypothesis that the Francisella tularensis AcrAB RND efflux system contributes to antimicrobial resistance and pathogenesis has been tested. A null mutation was generated in the gene encoding the AcrB RND efflux pump protein of the live vaccine strain of F. tularensis. The resulting mutant exhibited increased sensitivity to multiple antibiotics and antimicrobial compounds. Murine challenge experiments revealed that the acrB mutant was attenuated. Collectively these results suggest that the F. tularensis AcrAB RND efflux system encodes a multiple drug efflux system that is important for virulence.     

Characterization and expression analysis of Staphylococcus aureus pathogenicity island 3. Implications for the evolution of staphylococcal pathogenicity islands

We describe the complete sequence of the 15.9-kb staphylococcal pathogenicity island 3 encoding staphylococcal enterotoxin serotypes B, K, and Q. The island, which meets the generally accepted definition of pathogenicity islands, contains 24 open reading frames potentially encoding proteins of more than 50 amino acids, including an apparently functional integrase. The element is bordered by two 17-bp direct repeats identical to those found flanking staphylococcal pathogenicity island 1. The island has extensive regions of homology to previously described pathogenicity islands, particularly staphylococcal pathogenicity islands 1 and bov. The expression of 22 of the 24 open reading frames contained on staphylococcal pathogenicity island 3 was detected either in vitro during growth in a laboratory medium or serum or in vivo in a rabbit model of toxic shock syndrome using DNA microarrays. The effect of oxygen tension on staphylococcal pathogenicity island 3 gene expression was also examined. By comparison with the known staphylococcal pathogenicity islands in the context of gene expression described here, we propose a model of pathogenicity island origin and evolution involving specialized transduction events and addition, deletion, or recombination of pathogenicity island "modules."     

Interaction between bile acids and staphylococcal polysaccharide: inhibition of capsule formation in encapsulated mutant strains (taurine+, taurine-) of Staphylococcus aureus

In a previous paper, we showed that bile acid derivatives inhibit capsule formation as well as taurine biosynthesis in a taurine+ (Tau+) encapsulated strain of Staphylococcus aureus. In the present study, binding of [14C]cholic acid [( 14C]CA) and [14C]taurocholic acid [( 14C]TA) to the staphylococcal polysaccharide antigen (SPA) of the capsular fraction was examined. The bile acids were found to bind with SPA via taurine of the Tau+ cells. [14C]CA bound with the SPA fraction of the Tau+ strain within 10-30 min, whereas 60-120 min was required in the binding of [14C]TA. Various bile acids competed with cholic acid binding to Tau+ cells which was shown by the inhibition of binding with cholic acid or taurocholic acid but not with glycholic acid. Binding of bile acid derivatives to a Tau- encapsulated mutant or to capsular material from this mutant was not observed.     

Whole-genome sequencing of staphylococcus haemolyticus uncovers the extreme plasticity of its genome and the evolution of human-colonizing staphylococcal species

Staphylococcus haemolyticus is an opportunistic bacterial pathogen that colonizes human skin and is remarkable for its highly antibiotic-resistant phenotype. We determined the complete genome sequence of S.haemolyticus to better understand its pathogenicity and evolutionary relatedness to the other staphylococcal species. A large proportion of the open reading frames in the genomes of S.haemolyticus, Staphylococcus aureus, and Staphylococcus epidermidis were conserved in their sequence and order on the chromosome. We identified a region of the bacterial chromosome just downstream of the origin of replication that showed little homology among the species but was conserved among strains within a species. This novel region, designated the "oriC environ," likely contributes to the evolution and differentiation of the staphylococcal species, since it was enriched for species-specific nonessential genes that contribute to the biological features of each staphylococcal species. A comparative analysis of the genomes of S.haemolyticus, S.aureus, and S.epidermidis elucidated differences in their biological and genetic characteristics and pathogenic potentials. We identified as many as 82 insertion sequences in the S.haemolyticus chromosome that probably mediated frequent genomic rearrangements, resulting in phenotypic diversification of the strain. Such rearrangements could have brought genomic plasticity to this species and contributed to its acquisition of antibiotic resistance.     

Side effects of antibiotics in patients with thermal burns]

The possibilities of antibacterial therapy in the clinics of burn diseases has at present decreased because of increasing microflora resistance to antibiotics. This phenomenon is one of the most often causes of antibacterial drug side effects in burn patients. For control of infections complications in burn patients which are most often lethal it is necessary to use biologically active subtance, such as prodigiozan and lysozime in addition to the directed antibiotic therapy. The use of specific antitoxic antistaphylococcal drugs, such as antistaphylococcal plasma and antistaphylococcal gamma-globulin in combination with the antibiotics of the direct action provided effective control of infectious complications and sepsis of staphylococcal genesis in burn patients. Decamine proved to be effective along with the usual use of nystatin in cases with dysbacteriosis as a result of the antibiotic side effects. In the patients treated with decamine the sings of candidosis disappeared by the 5th--7th day. Therefore, for decreasing the side effects of antibiotics in the clinics of burn patients it is expedient to use antibiotics in combination with the biologically active and immune preparations which increases the efficacy of antibiotic therapy, impfoves the treatment results and decreases the antibiotic side effects.     

Isolation and characterization of a high molecular weight antibiotic produced by a marine bacterium

Many marine bacteria demonstrate antibiotic activity against organisms of terrestrial origin. Low molecular weight antibiotics have been extracted and, in some cases, purified, but few attempts have been made to isolate high molecular weight antibiotics produced by marine bacteria. In the study reported here, a high molecular weight antibiotic was extracted from whole cells ofAlteromonas strain P18 (NCMB 1890) grown on 2216E medium. Purification included ammonium sulfate precipitation, ultracentrifugation, chromatography on DEAE cellulose, and gel filtration on Ultrogel. A rapid method for measuring specific activity of the antibiotic was developed.