Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 37
Filter
1.
J Infect Dis ; 215(5): 703-712, 2017 03 01.
Article in English | MEDLINE | ID: mdl-28007922

ABSTRACT

Background: Increasing antibiotic resistance warrants therapeutic alternatives. Here we investigated the efficacy of bacteriophage-therapy (phage) alone or combined with antibiotics against experimental endocarditis (EE) due to Pseudomonas aeruginosa, an archetype of difficult-to-treat infection. Methods: In vitro fibrin clots and rats with aortic EE were treated with an antipseudomonas phage cocktail alone or combined with ciprofloxacin. Phage pharmacology, therapeutic efficacy, and resistance were determined. Results: In vitro, single-dose phage therapy killed 7 log colony-forming units (CFUs)/g of fibrin clots in 6 hours. Phage-resistant mutants regrew after 24 hours but were prevented by combination with ciprofloxacin (2.5 × minimum inhibitory concentration). In vivo, single-dose phage therapy killed 2.5 log CFUs/g of vegetations in 6 hours (P < .001 vs untreated controls) and was comparable with ciprofloxacin monotherapy. Moreover, phage/ciprofloxacin combinations were highly synergistic, killing >6 log CFUs/g of vegetations in 6 hours and successfully treating 64% (n = 7/11) of rats. Phage-resistant mutants emerged in vitro but not in vivo, most likely because resistant mutations affected bacterial surface determinants important for infectivity (eg, the pilT and galU genes involved in pilus motility and LPS formation). Conclusions: Single-dose phage therapy was active against P. aeruginosa EE and highly synergistic with ciprofloxacin. Phage-resistant mutants had impaired infectivity. Phage-therapy alone or combined with antibiotics merits further clinical consideration.


Subject(s)
Anti-Bacterial Agents/pharmacology , Endocarditis/therapy , Phage Therapy/methods , Pseudomonas Infections/therapy , Pseudomonas aeruginosa/drug effects , Animals , Ciprofloxacin/pharmacology , Drug Resistance, Multiple, Bacterial , Endocarditis/microbiology , Female , Microbial Sensitivity Tests , Pseudomonas aeruginosa/pathogenicity , Rats , Rats, Wistar , Virulence
2.
Antimicrob Agents Chemother ; 60(9): 5349-56, 2016 09.
Article in English | MEDLINE | ID: mdl-27353266

ABSTRACT

In this study, silver/copper (Ag/Cu)-coated catheters were investigated for their efficacy in preventing methicillin-resistant Staphylococcus aureus (MRSA) infection in vitro and in vivo Ag and Cu were sputtered (67/33% atomic ratio) on polyurethane catheters by direct-current magnetron sputtering. In vitro, Ag/Cu-coated and uncoated catheters were immersed in phosphate-buffered saline (PBS) or rat plasma and exposed to MRSA ATCC 43300 at 10(4) to 10(8) CFU/ml. In vivo, Ag/Cu-coated and uncoated catheters were placed in the jugular vein of rats. Directly after, MRSA (10(7) CFU/ml) was inoculated in the tail vein. Catheters were removed 48 h later and cultured. In vitro, Ag/Cu-coated catheters preincubated in PBS and exposed to 10(4) to 10(7) CFU/ml prevented the adherence of MRSA (0 to 12% colonization) compared to uncoated catheters (50 to 100% colonization; P < 0.005) and Ag/Cu-coated catheters retained their activity (0 to 20% colonization) when preincubated in rat plasma, whereas colonization of uncoated catheters increased (83 to 100%; P < 0.005). Ag/Cu-coating protection diminished with 10(8) CFU/ml in both PBS and plasma (50 to 100% colonization). In vivo, Ag/Cu-coated catheters reduced the incidence of catheter infection compared to uncoated catheters (57% versus 79%, respectively; P = 0.16) and bacteremia (31% versus 68%, respectively; P < 0.05). Scanning electron microscopy of explanted catheters suggests that the suboptimal activity of Ag/Cu catheters in vivo was due to the formation of a dense fibrin sheath over their surface. Ag/Cu-coated catheters thus may be able to prevent MRSA infections. Their activity might be improved by limiting plasma protein adsorption on their surfaces.


Subject(s)
Anti-Infective Agents/pharmacology , Bacteremia/prevention & control , Catheters, Indwelling/microbiology , Coated Materials, Biocompatible/pharmacology , Copper/pharmacology , Silver/pharmacology , Staphylococcal Infections/prevention & control , Adsorption , Animals , Bacteremia/microbiology , Colony Count, Microbial , Fibrin/chemistry , Jugular Veins , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/growth & development , Nanoparticles/chemistry , Nanoparticles/ultrastructure , Polyurethanes/chemistry , Rats , Staphylococcal Infections/microbiology
3.
Appl Microbiol Biotechnol ; 100(13): 5945-53, 2016 Jul.
Article in English | MEDLINE | ID: mdl-27020284

ABSTRACT

Using direct current magnetron sputtering (DCMS), we generated flexible copper polyester surfaces (Cu-PES) and investigated their antimicrobial activity against a range of multidrug-resistant (MDR) pathogens including eight Gram-positive isolates (three methicillin-resistant Staphylococcus aureus [MRSA], four vancomycin-resistant enterococci, one methicillin-resistant Staphylococcus epidermidis) and four Gram-negative strains (one extended-spectrum ß-lactamase-producing [ESBL] Escherichia coli, one ESBL Klebsiella pneumoniae, one imipenem-resistant Pseudomonas aeruginosa, and one ciprofloxacin-resistant Acinetobacter baumannii). Bactericidal activity (≥3 log10 CFU reduction of the starting inoculum) was reached within 15-30 min exposure to Cu-PES. Antimicrobial activity of Cu-PES persisted in the absence of oxygen and against both Gram-positive and Gram-negative bacteria containing elevated levels of catalases, indicating that reactive oxygen species (ROS) do not play a primary role in the killing process. The decrease in cell viability of MRSA ATCC 43300 and Enterococcus faecalis V583 correlated with the progressive loss of cytoplasmic membrane integrity both under aerobic and anaerobic conditions, suggesting that Cu-PES mediated killing is primarily induced by disruption of the cytoplasmic membrane function. Overall, we here present novel antimicrobial copper surfaces with improved stability and sustainability and provide further insights into their mechanism of killing.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Infections/microbiology , Copper/pharmacology , Drug Resistance, Bacterial , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Anti-Bacterial Agents/chemistry , Copper/chemistry , Drug Evaluation, Preclinical , Humans , Microbial Sensitivity Tests , Nanoparticles/chemistry
4.
Antimicrob Agents Chemother ; 59(4): 2435-8, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25605361

ABSTRACT

Methicillin-resistant Staphylococcus aureus (MRSA) carrying the mecC gene (mecC-MRSA) exhibited at 37°C MICs of oxacillin close to those of methicillin-susceptible S. aureus (MSSA). We investigated whether at this temperature, mecC-MRSA strains respond to flucloxacillin treatment like MSSA strains, using a rat model of endocarditis. Flucloxacillin (human-like kinetics of 2 g intravenously every 6 h) cured 80 to 100% of aortic vegetations infected with five different mecC-MRSA strains. These results suggest that mecC-MRSA infections may successfully respond to treatment with ß-lactams.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Endocarditis, Bacterial/drug therapy , Floxacillin/therapeutic use , Methicillin-Resistant Staphylococcus aureus/genetics , Staphylococcal Infections/drug therapy , Animals , Anti-Bacterial Agents/administration & dosage , Aorta/microbiology , Cefoxitin/pharmacology , Chromatography, Micellar Electrokinetic Capillary , Endocarditis, Bacterial/microbiology , Floxacillin/administration & dosage , Infusion Pumps , Microbial Sensitivity Tests , Oxacillin/pharmacology , Rats , Staphylococcal Infections/microbiology , Temperature
5.
Antimicrob Agents Chemother ; 56(8): 4511-5, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22644033

ABSTRACT

We describe 3 patients with left-sided staphylococcal endocarditis (1 with methicillin-susceptible Staphylococcus aureus [MSSA] prosthetic aortic valve endocarditis and 2 with methicillin-resistant S. aureus [MRSA] native-valve endocarditis) who were successfully treated with high-dose intravenous daptomycin (10 mg/kg/day) plus fosfomycin (2 g every 6 h) for 6 weeks. This combination was tested in vitro against 7 MSSA, 5 MRSA, and 2 intermediately glycopeptide-resistant S. aureus isolates and proved to be synergistic against 11 (79%) strains and bactericidal against 8 (57%) strains. This combination deserves further clinical study.


Subject(s)
Anti-Bacterial Agents/administration & dosage , Daptomycin/administration & dosage , Endocarditis, Bacterial/drug therapy , Fosfomycin/administration & dosage , Methicillin-Resistant Staphylococcus aureus/drug effects , Staphylococcal Infections/drug therapy , Aged , Anti-Bacterial Agents/adverse effects , Anti-Bacterial Agents/therapeutic use , Daptomycin/adverse effects , Daptomycin/therapeutic use , Drug Synergism , Drug Therapy, Combination , Female , Fosfomycin/adverse effects , Fosfomycin/therapeutic use , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Staphylococcal Infections/microbiology
6.
J Antimicrob Chemother ; 67(3): 652-60, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22167243

ABSTRACT

OBJECTIVES: Laboratory detection of vancomycin-intermediate Staphylococcus aureus (VISA) and their heterogeneous VISA (hVISA) precursors is difficult. Thus, it is possible that vancomycin failures against supposedly vancomycin-susceptible S. aureus are due to undiagnosed VISA or hVISA. We tested this hypothesis in experimental endocarditis. METHODS: Rats with aortic valve infection due to the vancomycin-susceptible (MIC 2 mg/L), methicillin-resistant S. aureus M1V2 were treated for 2 days with doses of vancomycin that mimicked the pharmacokinetics seen in humans following intravenous administration of 1 g of the drug every 12 h. Half of the treated animals were killed 8 h after treatment arrest and half 3 days thereafter. Population analyses were done directly on vegetation homogenates or after one subculture in drug-free medium to mimic standard diagnostic procedures. RESULTS: Vancomycin cured 14 of 26 animals (54%; P<0.05 versus controls) after 2 days of treatment. When vegetation homogenates were plated directly on vancomycin-containing plates, 6 of 13 rats killed 8 h after treatment arrest had positive cultures, 1 of which harboured hVISA. Likewise, 6 of 13 rats killed 3 days thereafter had positive valve cultures, 5 of which harboured hVISA. However, one subculture of vegetations in drug-free broth was enough to revert all the hVISA phenotypes to the susceptible pattern of the parent. Thus, vancomycin selected for hVISA during therapy of experimental endocarditis due to vancomycin-susceptible S. aureus. These hVISA were associated with vancomycin failure. The hVISA phenotype persisted in vivo, even after vancomycin arrest, but was missed in vitro after a single passage of the vegetation homogenate on drug-free medium. CONCLUSIONS: hVISA might escape detection in clinical samples if they are subcultured before susceptibility tests.


Subject(s)
Anti-Bacterial Agents/administration & dosage , Bacteriological Techniques/methods , Endocarditis, Bacterial/drug therapy , Staphylococcal Infections/drug therapy , Staphylococcus aureus/drug effects , Vancomycin Resistance , Vancomycin/administration & dosage , Animals , Disease Models, Animal , Endocarditis, Bacterial/microbiology , Injections, Intravenous , Rats , Selection, Genetic , Sensitivity and Specificity , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification
7.
J Infect Dis ; 204(12): 1960-70, 2011 Dec 15.
Article in English | MEDLINE | ID: mdl-22043018

ABSTRACT

BACKGROUND: Streptococcus gallolyticus is a causative agent of infective endocarditis associated with colon cancer. Genome sequence of strain UCN34 revealed the existence of 3 pilus loci (pil1, pil2, and pil3). Pili are long filamentous structures playing a key role as adhesive organelles in many pathogens. The pil1 locus encodes 2 LPXTG proteins (Gallo2178 and Gallo2179) and 1 sortase C (Gallo2177). Gallo2179 displaying a functional collagen-binding domain was referred to as the adhesin, whereas Gallo2178 was designated as the major pilin. METHODS: S. gallolyticus UCN34, Pil1(+) and Pil1(-), expressing various levels of pil1, and recombinant Lactococcus lactis strains, constitutively expressing pil1, were studied. Polyclonal antibodies raised against the putative pilin subunits Gallo2178 and Gallo2179 were used in immunoblotting and immunogold electron microscopy. The role of pil1 was tested in a rat model of endocarditis. RESULTS: We showed that the pil1 locus (gallo2179-78-77) forms an operon differentially expressed among S. gallolyticus strains. Short pilus appendages were identified both on the surface of S. gallolyticus UCN34 and recombinant L. lactis-expressing pil1. We demonstrated that Pil1 pilus is involved in binding to collagen, biofilm formation, and virulence in experimental endocarditis. CONCLUSIONS: This study identifies Pil1 as the first virulence factor characterized in S. gallolyticus.


Subject(s)
Biofilms , Endocarditis/microbiology , Fimbriae Proteins/genetics , Fimbriae, Bacterial/genetics , Genomic Islands/genetics , Streptococcus/genetics , Streptococcus/physiology , Animals , Collagen/metabolism , Female , Fimbriae Proteins/metabolism , Fimbriae, Bacterial/metabolism , Genetic Loci/genetics , Operon/genetics , Rats , Rats, Wistar , Streptococcus/metabolism , Streptococcus/pathogenicity , Virulence Factors/genetics , Virulence Factors/metabolism
8.
J Am Heart Assoc ; 11(3): e023080, 2022 02.
Article in English | MEDLINE | ID: mdl-35043655

ABSTRACT

Background The potential of phage therapy for the treatment of endovascular Staphylococcus aureus infections remains to be evaluated. Methods and Results The efficacy of a phage cocktail combining Herelleviridae phage vB_SauH_2002 and Podoviriae phage 66 was evaluated against a methicillin-sensitive S. aureus strain in vitro and in vivo in a rodent model of experimental endocarditis. Six hours after bacterial challenge, animals were treated with (1) the phage cocktail. (2) subtherapeutic flucloxacillin dosage, (3) combination of the phage cocktail and flucloxacillin, or (4) saline. Bacterial loads in cardiac vegetations at 30 hours were the primary outcome. Secondary outcomes were phage loads at 30 hours in cardiac vegetations, blood, spleen, liver, and kidneys. We evaluated phage resistance 30 hours post infection in vegetations of rats under combination treatment. In vitro, phages synergized against S. aureus planktonic cells and the cocktail synergized with flucloxacillin to eradicated biofilms. In infected animals, the phage cocktail achieved bacteriostatic effect. The addition of low-dose flucloxacillin elevated bacterial suppression (∆ of -5.25 log10 colony forming unit/g [CFU/g] versus treatment onset, P<0.0001) and synergism was confirmed (∆ of -2.15 log10 CFU/g versus low-dose flucloxacillin alone, P<0.01). Importantly, 9/12 rats given the combination treatment had sterile vegetations at 30 hours. In vivo phage replication was partially suppressed by the antibiotic and selection of resistance to the Podoviridae component of the phage cocktail occurred. Plasma-mediated inhibition of phage killing activity was observed in vitro. Conclusions Combining phages with a low-dose standard of care antibiotic represents a promising strategy for the treatment of S. aureus infective endocarditis.


Subject(s)
Bacteriophages , Endocarditis, Bacterial , Endocarditis , Staphylococcal Infections , Animals , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Bacteriophages/physiology , Endocarditis/microbiology , Endocarditis, Bacterial/therapy , Floxacillin/pharmacology , Floxacillin/therapeutic use , Rats , Staphylococcal Infections/therapy , Staphylococcus aureus/physiology
9.
J Exp Med ; 201(10): 1627-35, 2005 May 16.
Article in English | MEDLINE | ID: mdl-15897276

ABSTRACT

The expression of Staphylococcus aureus adhesins in Lactococcus lactis identified clumping factor A (ClfA) and fibronectin-binding protein A (FnBPA) as critical for valve colonization in rats with experimental endocarditis. This study further analyzed their role in disease evolution. Infected animals were followed for 3 d. ClfA-positive lactococci successfully colonized damaged valves, but were spontaneously eradicated over 48 h. In contrast, FnBPA-positive lactococci progressively increased bacterial titers in vegetations and spleens. At imaging, ClfA-positive lactococci were restricted to the vegetations, whereas FnBPA-positive lactococci also invaded the adjacent endothelium. This reflected the capacity of FnBPA to trigger cell internalization in vitro. Because FnBPA carries both fibrinogen- and fibronectin-binding domains, we tested the role of these functionalities by deleting the fibrinogen-binding domain of FnBPA and supplementing it with the fibrinogen-binding domain of ClfA in cis or in trans. Deletion of the fibrinogen-binding domain of FnBPA did not alter fibronectin binding and cell internalization in vitro. However, it totally abrogated valve infectivity in vivo. This ability was restored in cis by inserting the fibrinogen-binding domain of ClfA into truncated FnBPA, and in trans by coexpressing full-length ClfA and truncated FnBPA on two separate plasmids. Thus, fibrinogen and fibronectin binding could cooperate for S. aureus valve colonization and endothelial invasion in vivo.


Subject(s)
Adhesins, Bacterial/metabolism , Bacterial Adhesion/genetics , Coagulase/metabolism , Endocarditis, Bacterial/microbiology , Heart Valves/microbiology , Staphylococcal Infections/microbiology , Staphylococcus aureus/pathogenicity , Adhesins, Bacterial/genetics , Animals , Coagulase/genetics , Endocarditis, Bacterial/metabolism , Endocarditis, Bacterial/pathology , Endothelium, Vascular/metabolism , Endothelium, Vascular/microbiology , Endothelium, Vascular/pathology , Female , Fibrinogen/metabolism , Fibronectins/metabolism , Heart Valves/metabolism , Heart Valves/pathology , Lactococcus lactis/genetics , Lactococcus lactis/pathogenicity , Protein Binding , Protein Structure, Tertiary/genetics , Rats , Rats, Wistar , Sequence Deletion , Spleen/metabolism , Spleen/microbiology , Spleen/pathology , Staphylococcal Infections/metabolism , Staphylococcal Infections/pathology , Staphylococcus aureus/genetics
10.
Infect Immun ; 78(4): 1711-6, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20065030

ABSTRACT

Adherence to fibrinogen and fibronectin plays a crucial role in Staphylococcus aureus experimental endocarditis. Previous genetic studies have shown that infection and carriage isolates do not systematically differ in their virulence-related genes, including genes conferring adherence, such as clfA and fnbA. We set out to determine the range of adherence phenotypes in carriage isolates of S. aureus, to compare the adherence of these isolates to the adherence of infection isolates, and to determine the relationship between adherence and infectivity in a rat model of experimental endocarditis. A total of 133 healthy carriage isolates were screened for in vitro adherence to fibrinogen and fibronectin, and 30 isolates were randomly chosen for further investigation. These 30 isolates were compared to 30 infective endocarditis isolates and 30 blood culture isolates. The infectivities of the carriage isolates, which displayed either extremely low or high adherence to fibrinogen and fibronectin, were tested using a rat model of experimental endocarditis. The levels of adherence to both fibrinogen and fibronectin were very similar for isolates from healthy carriers and members of the two groups of infection isolates. All three groups of isolates showed a wide range of adherence to fibrinogen and fibronectin. Moreover, the carriage isolates that showed minimal adherence and the carriage isolates that showed strong adherence had the same infectivity in experimental endocarditis. Adherence was proven to be important for pathogenesis in experimental endocarditis, but even the least adherent carriage strains had the ability to induce infection. We discuss the roles of differential gene expression, human host factors, and gene redundancy in resolving this apparent paradox.


Subject(s)
Bacterial Adhesion , Carrier State/microbiology , Endocarditis, Bacterial/microbiology , Fibrinogen/metabolism , Fibronectins/metabolism , Staphylococcal Infections/microbiology , Staphylococcus aureus/pathogenicity , Adult , Animals , Disease Models, Animal , Gene Expression , Genetic Variation , Humans , Phenotype , Rats , Staphylococcus aureus/isolation & purification , Staphylococcus aureus/physiology , Young Adult
11.
Antimicrob Agents Chemother ; 53(9): 3635-41, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19564362

ABSTRACT

Iclaprim is a novel diaminopyrimidine antibiotic that is active against methicillin-resistant Staphylococcus aureus (MRSA). However, it is known that the activity of diaminopyrimidines against S. aureus is antagonized by thymidine through uptake and conversion to thymidylate by thymidine kinase. Unlike with humans, for whom thymidine levels are low, thymidine levels in rodents are high, thus precluding the accurate evaluation of iclaprim efficacy in animal models. We have studied the bactericidal activity of iclaprim against an isogenic pair of MRSA isolates, the wild-type parent AW6 and its thymidine kinase-deficient mutant AH1252, in an in vitro fibrin clot model. Clots, which were aimed at mimicking vegetation structure, were made from human or rat plasma containing either the parent AW6 or the mutant AH1252, and they were exposed to homologous serum supplemented with iclaprim (3.5 microg/ml), trimethoprim-sulfamethoxazole (TMP-SMX; 8/40 microg/ml), vancomycin (40 microg/ml), or saline, each of which was added one time for 48 h. In rat clots, iclaprim and TMP-SMX were bacteriostatic against the parent, AW6. In contrast, they were bactericidal (> or = 3 log10 CFU/clot killing of the original inoculum) against the mutant AH1252. Vancomycin was the most active drug against AW6 (P < 0.05), but it showed an activity similar those of iclaprim and TMP-SMX against AH1252. In human clots, iclaprim was bactericidal against both AW6 and AH1252 strains and was as effective as TMP-SMX and vancomycin (P > 0.05). Future studies of animals using simulated human kinetics of iclaprim and thymidine kinase-deficient MRSA, which eliminate the thymidine-induced confounding effect, are warranted to support the use of iclaprim in the treatment of severe MRSA infections in humans.


Subject(s)
Anti-Bacterial Agents/pharmacology , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/genetics , Pyrimidines/pharmacology , Thymidine Kinase/genetics , Animals , Humans , Microbial Sensitivity Tests , Mutation , Rats , Thymidine Kinase/physiology , Trimethoprim, Sulfamethoxazole Drug Combination/pharmacology , Vancomycin/pharmacology
12.
J Med Microbiol ; 58(Pt 7): 863-866, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19498205

ABSTRACT

Tissue microarray technology was used to establish immunohistochemistry protocols and to determine the specificity of new antisera against various Chlamydia-like bacteria for future use on formalin-fixed and paraffin-embedded tissues. The antisera exhibited strong reactivity against autologous antigen and closely related heterologous antigen, but no cross-reactivity with distantly related species.


Subject(s)
Antibodies, Bacterial/immunology , Chlamydiales/immunology , Immunohistochemistry , Microarray Analysis/methods , Acanthamoeba castellanii/microbiology , Animals , Antibody Specificity , Chlorocebus aethiops , Cross Reactions , Immune Sera/immunology , Mice , Vero Cells
13.
Infect Immun ; 76(8): 3824-31, 2008 Aug.
Article in English | MEDLINE | ID: mdl-18541660

ABSTRACT

Staphylococcus aureus experimental endocarditis relies on sequential fibrinogen binding (for valve colonization) and fibronectin binding (for endothelial invasion) conferred by peptidoglycan-attached adhesins. Fibronectin-binding protein A (FnBPA) reconciles these two properties--as well as elastin binding--and promotes experimental endocarditis by itself. Here we attempted to delineate the minimal subdomain of FnBPA responsible for fibrinogen and fibronectin binding, cell invasion, and in vivo endocarditis. A large library of truncated constructs of FnBPA was expressed in Lactococcus lactis and tested in vitro and in animals. A 127-amino-acid subdomain spanning the hinge of the FnBPA fibrinogen-binding and fibronectin-binding regions appeared necessary and sufficient to confer the sum of these properties. Competition with synthetic peptides could not delineate specific fibrinogen- and fibronectin-binding sites, suggesting that dual binding arose from protein folding, irrespective of clearly defined binding domains. Moreover, coexpressing the 127-amino-acid subdomain with remote domains of FnBPA further increased fibrinogen binding by > or =10 times, confirming the importance of domain interactions for binding efficacy. In animals, fibrinogen binding (but not fibronectin binding) was significantly associated with endocarditis induction, whereas both fibrinogen binding and fibronectin binding were associated with disease severity. Moreover, fibrinogen binding also combined with fibronectin binding to synergize the invasion of cultured cell lines significantly, a feature correlating with endocarditis severity. Thus, while fibrinogen binding and fibronectin binding were believed to act sequentially in colonization and invasion, they appeared unexpectedly intertwined in terms of both functional anatomy and pathogenicity (in endocarditis). This unforeseen FnBPA subtlety might bear importance for the development of antiadhesin strategies.


Subject(s)
Adhesins, Bacterial/metabolism , Endocarditis, Bacterial/microbiology , Endothelial Cells/microbiology , Staphylococcus aureus/pathogenicity , Virulence Factors/metabolism , Adhesins, Bacterial/genetics , Animals , Cattle , Cell Line , Female , Fibrinogen/metabolism , Fibronectins/metabolism , Gene Expression , Lactococcus lactis/genetics , Protein Binding , Protein Interaction Domains and Motifs , Protein Interaction Mapping , Rats , Sequence Deletion , Severity of Illness Index , Virulence Factors/genetics
14.
J Antimicrob Chemother ; 61(2): 371-4, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18033780

ABSTRACT

OBJECTIVES: To test the activity of tigecycline combined with 16 antimicrobials in vitro against 22 gram-positive and 55 gram-negative clinical isolates. METHODS: Antibiotic interactions were determined by chequerboard and time-kill methods. RESULTS: By chequerboard, of 891 organism-drug interactions tested, 97 (11%) were synergistic, 793 (89%) were indifferent and 1 (0.1%) was antagonistic. Among gram-positive pathogens, most synergisms occurred against Enterococcus spp. (7/11 isolates) with the tigecycline/rifampicin combination. No antagonism was detected. Among gram-negative organisms, synergism was observed mainly with trimethoprim/sulfamethoxazole against Serratia marcescens (5/5 isolates), Proteus spp. (2/5) and Stenotrophomonas maltophilia (2/5), with aztreonam against S. maltophilia (3/5), with cefepime and imipenem against Enterobacter cloacae (3/5), with ceftazidime against Morganella morganii (3/5), and with ceftriaxone against Klebsiella pneumoniae (3/5). The only case of antagonism occurred against one S. marcescens with the tigecycline/imipenem combination. Selected time-kill assays confirmed the bacteriostatic interactions observed by the chequerboard method. Moreover, they revealed a bactericidal synergism of tigecycline with piperacillin/tazobactam against one penicillin-resistant Streptococcus pneumoniae and with amikacin against Proteus vulgaris. CONCLUSIONS: Combinations of tigecycline with other antimicrobials produce primarily an indifferent response. Specific synergisms, especially against enterococci and problematic gram-negative isolates, might be worth investigating in in vitro models and/or in animal models simulating the human environment.


Subject(s)
Anti-Infective Agents/administration & dosage , Drug Resistance, Multiple, Bacterial/drug effects , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Minocycline/analogs & derivatives , Drug Resistance, Multiple, Bacterial/physiology , Drug Therapy, Combination , Gram-Negative Bacteria/physiology , Gram-Positive Bacteria/physiology , Humans , Microbial Sensitivity Tests/methods , Minocycline/administration & dosage , Tigecycline
15.
Thromb Haemost ; 118(7): 1230-1241, 2018 Jul.
Article in English | MEDLINE | ID: mdl-29909601

ABSTRACT

Adhesion of Staphylococcus aureus to endothelial cells (ECs) is paramount in infective endocarditis. Bacterial proteins such as clumping factor A (ClfA) and fibronectin binding protein A (FnbpA) mediate adhesion to EC surface molecules and (sub)endothelial matrix proteins including fibrinogen (Fg), fibrin, fibronectin (Fn) and von Willebrand factor (vWF). We studied the influence of shear flow and plasma on the binding of ClfA and FnbpA (including its sub-domains A, A16+, ABC, CD) to coverslip-coated vWF, Fg/fibrin, Fn or confluent ECs, making use of Lactococcus lactis, expressing these adhesins heterologously. Global adherence profiles were similar in static and flow conditions. In the absence of plasma, L. lactis-clfA binding to Fg increased with shear forces, whereas binding to fibrin did not. The degree of adhesion of L. lactis-fnbpA to EC-bound Fn and of L. lactis-clfA to EC-bound Fg, furthermore, was similar to that of L. lactis-clfA to coated vWF domain A1, in the presence of vWF-binding protein (vWbp). Yet, in plasma, L. lactis-clfA adherence to activated EC-vWF/vWbp dropped over 10 minutes by 80% due to vWF-hydrolysis by a disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13 and that of L. lactis-fnbpA likewise by > 70% compared to the adhesion in absence of plasma. In contrast, plasma Fg supported high L. lactis-clfA binding to resting and activated ECs. Or, in plasma S. aureus adhesion to active endothelium occurs mainly via two complementary pathways: a rapid but short-lived vWF/vWbp pathway and a stable integrin-coupled Fg-pathway. Hence, the pharmacological inhibition of ClfA-Fg interactions may constitute a valuable additive treatment in infective endocarditis.


Subject(s)
ADAMTS13 Protein/blood , Bacterial Adhesion , Coagulase/metabolism , Endocarditis, Bacterial/microbiology , Human Umbilical Vein Endothelial Cells/microbiology , Plasma/enzymology , Staphylococcus aureus/metabolism , Adhesins, Bacterial/genetics , Adhesins, Bacterial/metabolism , Cells, Cultured , Coagulase/genetics , Endocarditis, Bacterial/blood , Fibrin/metabolism , Fibrinogen , Fibronectins/metabolism , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Lactococcus lactis/genetics , Lactococcus lactis/metabolism , Protein Binding , Protein Interaction Domains and Motifs , Staphylococcus aureus/genetics , Stress, Mechanical , von Willebrand Factor/metabolism
16.
J Med Microbiol ; 56(Pt 8): 1017-1024, 2007 Aug.
Article in English | MEDLINE | ID: mdl-17644707

ABSTRACT

The potential pathogenicity of selected (potentially) probiotic and clinical isolates of Lactobacillus rhamnosus and Lactobacillus paracasei was investigated in a rat model of experimental endocarditis. In addition, adhesion properties of the lactobacilli for fibrinogen, fibronectin, collagen and laminin, as well as the killing activity of the platelet-microbicidal proteins fibrinopeptide A (FP-A) and connective tissue activating peptide 3 (CTAP-3), were assessed. The 90 % infective dose (ID(90)) of the L. rhamnosus endocarditis isolates varied between 10(6) and 10(7) c.f.u., whereas four of the six (potentially) probiotic L. rhamnosus isolates showed an ID(90) that was at least 10-fold higher (10(8) c.f.u.) (P<0.001). In contrast, the two other probiotic L. rhamnosus isolates exhibited an ID(90) (10(6) and 10(7) c.f.u.) comparable to the ID(90) of the clinical isolates of this species investigated (P>0.05). Importantly, these two probiotic isolates shared the same fluorescent amplified fragment length polymorphism cluster type as the clinical isolate showing the lowest ID(90) (10(6) c.f.u.). L. paracasei tended to have a lower infectivity than L. rhamnosus (ID(90) of 10(7) to > or =10(8) c.f.u.). All isolates had comparable bacterial counts in cardiac vegetations (P>0.05). Except for one L. paracasei strain adhering to all substrates, all tested lactobacilli adhered only weakly or not at all. The platelet peptide FP-A did not show any microbicidal activity against the tested lactobacilli, whereas CTAP-3 killed the majority of the isolates. In general, these results indicate that probiotic lactobacilli display a lower infectivity in experimental endocarditis compared with true endocarditis pathogens. However, the difference in infectivity between L. rhamnosus endocarditis and (potentially) probiotic isolates could not be explained by differences in adherence or platelet microbicidal protein susceptibility. Other disease-promoting factors may exist in these organisms and warrant further investigation.


Subject(s)
Disease Models, Animal , Endocarditis, Bacterial/microbiology , Lactobacillus/pathogenicity , Animals , Bacterial Adhesion/drug effects , Dose-Response Relationship, Drug , Female , Fibrinopeptide A/pharmacology , Humans , Lactobacillus/drug effects , Lactobacillus/physiology , Lacticaseibacillus rhamnosus/drug effects , Lacticaseibacillus rhamnosus/pathogenicity , Lacticaseibacillus rhamnosus/physiology , Peptides/immunology , Peptides/pharmacology , Probiotics , Rats , Rats, Wistar , Species Specificity , Virulence
17.
J Photochem Photobiol B ; 174: 229-234, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28802173

ABSTRACT

Candida spp. are able to survive on hospital surfaces and causes healthcare-associated infections (HCAIs). Since surface cleaning and disinfecting interventions are not totally effective to eliminate Candida spp., new approaches should be devised. Copper (Cu) has widely recognized antifungal activity and the use of Cu-sputtered surfaces has recently been proposed to curb the spread of HCAIs. Moreover, the activity of Cu under the action of actinic light remains underexplored. We investigated the antifungal activity of Cu-sputtered polyester surfaces (Cu-PES) against azole-resistant Candida albicans and Candida glabrata under dark and low intensity visible light irradiation (4.65mW/cm2). The surface properties of Cu-PES photocatalysts were characterized by diffuse reflectance spectroscopy (DRS) and X-ray fluorescence (XRF). Under dark, Cu-PES showed a fungicidal activity (≥3log10CFU reduction of the initial inoculum) against both C. albicans DSY296 and C. glabrata DSY565 leading to a reduction of the starting inoculum of 3.1 and 3.0log10CFU, respectively, within 60min of exposure. Under low intensity visible light irradiation, Cu-PES exhibited an accelerated fungicidal activity against both strains with a reduction of 3.0 and 3.4log10CFU, respectively, within 30min of exposure. This effect was likely due to the semiconductor Cu2O/CuO charge separation. The decrease in cell viability of the two Candida strains under dark and light conditions correlated with the progressive loss of membrane integrity. These results indicate that Cu-PES represent a promising strategy for decreasing the colonization of surfaces by yeasts and that actinic light can improve its self-disinfecting activity.


Subject(s)
Antifungal Agents/pharmacology , Azoles/pharmacology , Candida albicans/drug effects , Candida albicans/radiation effects , Copper/pharmacology , Darkness , Drug Resistance, Fungal/drug effects , Antifungal Agents/chemistry , Candida albicans/cytology , Cell Membrane/drug effects , Cell Membrane/metabolism , Cell Membrane/radiation effects , Copper/chemistry , Drug Resistance, Fungal/radiation effects , Polyesters/chemistry , Surface Properties
18.
FEMS Immunol Med Microbiol ; 48(2): 215-7, 2006 Nov.
Article in English | MEDLINE | ID: mdl-17064277

ABSTRACT

During a 6-year period, we isolated three Abiotrophia defectiva, six Granulicatella adiacens and two G. 'para-adiacens' strains from clinical specimens. All A. defectiva strains were isolated from immunocompetent patients with endovascular infections, whereas the Granulicatella spp. strains were isolated from immunosuppressed patients with primary bacteremia. As the capacity of bacteria to adhere to the host extracellular matrix (ECM) has been implicated in the pathogenesis of endovascular infection, we investigated the ability of A. defectiva and Granulicatella spp. isolates to bind different ECM components immobilized in microtiter plates. Adherence tests showed a strong attachment of A. defectiva strains to fibronectin, whereas Granulicatella spp. strains were not adherent. The poor adherence of Granulicatella spp. strains to the ECM could be correlated with a lower propensity to induce endocarditis.


Subject(s)
Bacterial Adhesion/physiology , Endocarditis, Bacterial/microbiology , Fibronectins/metabolism , Streptococcal Infections/microbiology , Streptococcus/physiology , Extracellular Matrix/metabolism , Humans , Streptococcus/metabolism , Streptococcus/pathogenicity
19.
FEMS Immunol Med Microbiol ; 48(3): 419-23, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17087816

ABSTRACT

The ability to induce experimental endocarditis of biofilm-deficient mutants of Streptococcus gordonii was studied in an isogenic background. Strains were inactivated in either comD, fruK or pbp2b genes, which are involved in biofilm formation. These strains were clearly impaired (>75% reduction) in biofilm production in vitro. However, this did not result in a decreased severity of infection in vivo.


Subject(s)
Biofilms/growth & development , Endocarditis, Bacterial/microbiology , Streptococcal Infections/microbiology , Streptococcus/pathogenicity , Virulence/physiology , Animals , Bacterial Adhesion/genetics , Bacterial Adhesion/physiology , Disease Models, Animal , Female , Rats , Streptococcus/genetics , Virulence/genetics
20.
BMC Infect Dis ; 6: 9, 2006 Jan 20.
Article in English | MEDLINE | ID: mdl-16426445

ABSTRACT

BACKGROUND: Abiotrophia and Granulicatella species, previously referred to as nutritionally variant streptococci (NVS), are significant causative agents of endocarditis and bacteraemia. In this study, we reviewed the clinical manifestations of infections due to A. defectiva and Granulicatella species that occurred at our institution between 1998 and 2004. METHODS: The analysis included all strains of NVS that were isolated from blood cultures or vascular graft specimens. All strains were identified by 16S rRNA sequence analysis. Patients' medical charts were reviewed for each case of infection. RESULTS: Eleven strains of NVS were isolated during the 6-year period. Identification of the strains by 16S rRNA showed 2 genogroups: Abiotrophia defectiva (3) and Granulicatella adiacens (6) or "para-adiacens" (2). The three A. defectiva strains were isolated from immunocompetent patients with endovascular infections, whereas 7 of 8 Granulicatella spp. strains were isolated from immunosuppressed patients, mainly febrile neutropenic patients. We report the first case of "G. para-adiacens" bacteraemia in the setting of febrile neutropenia. CONCLUSION: We propose that Granulicatella spp. be considered as a possible agent of bacteraemia in neutropenic patients.


Subject(s)
Bacteremia/microbiology , Endocarditis, Bacterial/microbiology , Streptococcaceae/classification , Streptococcaceae/physiology , Adult , Female , Humans , Infant , Male , Microbial Sensitivity Tests , Middle Aged , Neutropenia/complications , Phylogeny , RNA, Ribosomal, 16S/genetics , Streptococcaceae/genetics
SELECTION OF CITATIONS
SEARCH DETAIL