scholarly journals Enterococcus faecalis Colonizes and Forms Persistent Biofilm Microcolonies on Undamaged Endothelial Surfaces in a Rabbit Endovascular Infection Model

FEMS Microbes ◽  
2021 ◽  
Author(s):  
Aaron M T Barnes ◽  
Kristi L Frank ◽  
Jennifer L Dale ◽  
Dawn A Manias ◽  
Jennifer L Powers ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Cell Attachment ◽  
Thrombus Formation ◽  
Bacterial Colonization ◽  
Surface Damage ◽  
Endothelial Damage ◽  
Bacterial Attachment ◽  
Infectious Endocarditis ◽  
Morphological Evidence ◽  
Infection Model

Abstract Infectious endocarditis is an uncommon disease with significant morbidity and mortality. The pathogenesis of infectious endocarditis has historically been described as a cascade of host-specific events beginning with endothelial damage and thrombus formation and followed by bacterial colonization of the nascent thrombus. Enterococcus faecalis is a Gram-positive commensal bacterial member of the gastrointestinal tract microbiota in most terrestrial animals and a leading cause of opportunistic biofilm-associated infections, including endocarditis. Here we provide evidence that E. faecalis can colonize the endocardial surface without pre-existing damage and in the absence of thrombus formation in a rabbit endovascular infection model. Using previously described light and scanning electron microscopy techniques, we show that inoculation of a well-characterized E. faecalis lab strain in the marginal ear vein of New Zealand White rabbits resulted in rapid colonization of the endocardium throughout the heart within four days of administration. Unexpectedly, ultrastructural imaging revealed that the microcolonies were firmly attached directly to the endocardium in areas without morphological evidence of gross tissue damage. Further, the attached bacterial aggregates were not associated with significant cellular components of coagulation or host extracellular matrix damage repair (i.e., platelets). These results suggest that the canonical model of mechanical surface damage as a prerequisite for bacterial attachment to host sub-endothelial components is not required. Furthermore, these findings are consistent with a model of initial establishment of stable, endocarditis-associated E. faecalis biofilm microcolonies that may provide a reservoir for the eventual valvular infection characteristic of clinical endocarditis. The similarities between the E. faecalis colonization and biofilm morphologies seen in this rabbit endovascular infection model and our previously published murine gastrointestinal colonization model indicate that biofilm production and common host cell attachment factors are conserved in disparate mammalian hosts under both commensal and pathogenic contexts.

An Insight into Surface Topographical Parameters and Bacterial Adhesion: A Case Study of Listeria monocytogenes Scott A Attachment on 304 Stainless Steel

2020 ◽  
Vol 83 (3) ◽  
pp. 426-433
Author(s):  
JAYANTI DAS ◽  
JENNIFER A. CHASE ◽  
MELISSA L. PARTYKA ◽  
EDWARD R. ATWILL ◽  
BARBARA LINKE
Keyword(s):  
Surface Roughness ◽  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Cell Attachment ◽  
Bacterial Colonization ◽  
Multivariate Regression Analysis ◽  
Bacterial Attachment ◽  
304 Stainless Steel ◽  
Attachment Behavior ◽  
Steel Surfaces

ABSTRACT Bacterial attachment on surfaces is an important biological and industrial concern. Many parameters affect cell attachment behavior, including surface roughness and other topographical features. An understanding of these relationships is critical in the light of recent outbreaks caused by foodborne bacteria. Postharvest packing lines have been identified as a potential source of cross-contamination with pathogens, which can cause subsequent foodborne illness. The objective of this article is to evaluate the influence of surface topographical features on bacterial attachment at various processing temperatures to determine the extent of bacterial colonization. Type 304 stainless steel surfaces and pathogenic Listeria monocytogenes Scott A were used for a detailed investigation. Two commonly used surface types, extruded and ground, were evaluated to determine differences in bacterial attachment on the same type of material. Fifteen surface topography parameters at three processing temperatures were studied to evaluate possible correlations with microbial attachment on these surfaces. Scanning electron microscopy, energy-dispersive X-ray spectroscopy, and confocal microscopy were used for both qualitative and quantitative analyses of surfaces. An analysis of variance and multivariate regression analysis were used to predict the attachment behavior of L. monocytogenes Scott A on stainless steel surfaces. Surface isotropy, average surface roughness, surface spacing, and processing temperatures were strongly correlated with bacterial attachment on 304 stainless steel material. HIGHLIGHTS

An Electron Microscopic Study of Platelet Thrombus Formation in the Rabbit with Particular Regard to 5-Hydroxytryptamine Release

1967 ◽  
Vol 18 (03/04) ◽  
pp. 592-604 ◽  
Author(s):  
H. R Baumgartner ◽  
J. P Tranzer ◽  
A Studer
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Vessel Wall ◽  
Thrombus Formation ◽  
Endothelial Damage ◽  
Electron Microscopic ◽  
Rabbit Ear ◽  
Basement Lamina ◽  
Platelet Thrombus

SummaryElectron microscopic and histologic examination of rabbit ear vein segments 4 and 30 min after slight endothelial damage have yielded the following findings :1. Platelets do not adhere to damaged endothelial cells.2. If the vessel wall is denuded of the whole endothelial cell, platelets adhere to the intimai basement lamina as do endothelial cells.3. The distance between adherent platelets as well as endothelial cells and intimai basement lamina measures 10 to 20 mµ, whereas the distance between aggregated platelets is 30 to 60 mµ.4. 5-hydroxytryptamine (5-HT) is released from platelets during viscous metamorphosis at least in part as 5-HT organelles.It should be noted that the presence of collagen fibers is not necessary for platelet thrombus formation in vivo.

Naturally Produced Extracellular Matrix Is an Excellent Substrate for Canine Endothelial Cell Proliferation and Resistance to Shear Stress on PTFE Vascular Grafts

1997 ◽  
Vol 78 (05) ◽  
pp. 1392-1398 ◽  
Author(s):  
A Schneider ◽  
M Chandra ◽  
G Lazarovici ◽  
I Vlodavsky ◽  
G Merin ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Cell Attachment ◽  
Thrombus Formation ◽  
Endothelial Cell Proliferation ◽  
Ptfe Graft ◽  
Vascular Prostheses ◽  
Endothelial Cell Attachment

SummaryPurpose: Successful development of a vascular prosthesis lined with endothelial cells (EC) may depend on the ability of the attached cells to resist shear forces after implantation. The present study was designed to investigate EC detachment from extracellular matrix (ECM) precoated vascular prostheses, caused by shear stress in vitro and to test the performance of these grafts in vivo. Methods: Bovine aortic endothelial cells were seeded inside untreated polytetrafluoro-ethylene (PTFE) vascular graft (10 X 0.6 cm), PTFE graft precoated with fibronectin (FN), or PTFE precoated with FN and a naturally produced ECM (106 cells/graft). Sixteen hours after seeding the medium was replaced and unattached cells counted. The strength of endothelial cell attachment was evaluated by subjecting the grafts to a physiologic shear stress of 15 dynes/cm2 for 1 h. The detached cells were collected and quantitated. PTFE or EC preseeded ECM coated grafts were implanted in the common carotid arteries of dogs. Results: While little or no differences were found in the extent of endothelial cell attachment to the various grafts (79%, 87% and 94% of the cells attached to PTFE, FN precoated PTFE, or FN+ECM precoated PTFE, respectively), the number of cells retained after a shear stress was significanly increased on ECM coated PTFE (20%, 54% and 85% on PTFE, FN coated PTFE, and FN+ECM coated PTFE, respectively, p <0.01). Implantation experiments in dogs revealed a significant increase in EC coverage and a reduced incidence of thrombus formation on ECM coated grafts that were seeded with autologous saphenous vein endothelial cells prior to implantation. Conclusion: ECM coating significantly increased the strength of endothelial cell attachment to vascular prostheses subjected to shear stress. The presence of adhesive macromolecules and potent endothelial cell growth promoting factors may render the ECM a promising substrate for vascular prostheses.

scholarly journals Pharmacodynamics of Daptomycin against Enterococcus faecium and Enterococcus faecalis in the Murine Thigh Infection Model

2018 ◽  
Vol 62 (10) ◽  
Author(s):  
James M. Kidd ◽  
Kamilia Abdelraouf ◽  
Tomefa E. Asempa ◽  
Romney M. Humphries ◽  
David P. Nicolau
Keyword(s):  
Enterococcus Faecalis ◽  
Enterococcus Faecium ◽  
Hill Equation ◽  
Infection Model ◽  
Free Drug Concentration ◽  
Time Curve ◽  
Content Type ◽  
Concentration Time ◽  
The Hill ◽  
Susceptibility Breakpoint

ABSTRACT The Clinical and Laboratory Standards Institute (CLSI) daptomycin MIC susceptibility breakpoint for the treatment of enterococcal infections is ≤4 μg/ml. However, patients receiving daptomycin for the treatment of infections caused by enterococci with MICs of ≤4 μg/ml may experience treatment failures. We assessed the pharmacodynamics of daptomycin against enterococci in a neutropenic murine thigh infection model and determined the exposures necessary for bacteriostasis and a 1-log10-CFU reduction of Enterococcus faecalis and Enterococcus faecium. We further characterized daptomycin efficacy at clinically achievable exposures. Six E. faecium and 6 E. faecalis isolates (daptomycin MICs, 0.5 to 32 μg/ml) were studied. Daptomycin was administered at various doses over 24 h to achieve area under the free drug concentration-time curve-to-MIC ratios (fAUC0–24/MIC) ranging from 1 to 148. Daptomycin regimens that simulate mean human exposures following doses of 6, 8, and 10 mg/kg of body weight/day were also studied. Efficacy was assessed by the differences in the number of log10 CFU per thigh at 24 h. The Hill equation was used to estimate the fAUC0–24/MIC required to achieve bacteriostasis and a 1-log10-CFU reduction. For E. faecium, a 1-log10-CFU reduction required an fAUC0–24/MIC of 12.9 (R2 = 0.71). For E. faecalis, a 1-log10-CFU reduction was not achieved, while the fAUC0–24/MIC required for stasis was 7.2 (R2 = 0.8). With a human-simulated regimen of 6 mg/kg/day, a 1-log10-CFU reduction was observed in 3/3 E. faecium isolates with MICs of <4 μg/ml and 0/3 E. faecium isolates with MICs of ≥4 μg/ml; however, a 1-log10-CFU reduction was not achieved for any of the 6 E. faecalis isolates. These results, alongside clinical data, prompt a reevaluation of the current breakpoint.

scholarly journals Pseudomonas aeruginosa las and rhl quorum-sensing systems are important for infection and inflammation in a rat prostatitis model

Microbiology ◽  
2009 ◽  
Vol 155 (8) ◽  
pp. 2612-2619 ◽  
Author(s):  
Lisa K. Nelson ◽  
Genevieve H. D'Amours ◽  
Kimberley M. Sproule-Willoughby ◽  
Douglas W. Morck ◽  
Howard Ceri
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Tissue Damage ◽  
Inflammatory Markers ◽  
Bacterial Colonization ◽  
Opportunistic Pathogen ◽  
Infection Model ◽  
Chronic Infections ◽  
Strain Pao1 ◽  
Rat Prostate

Pseudomonas aeruginosa frequently acts as an opportunistic pathogen of mucosal surfaces; yet, despite causing aggressive prostatitis in some men, its role as a pathogen in the prostate has not been investigated. Consequently, we developed a Ps. aeruginosa infection model in the rat prostate by instilling wild-type (WT) Ps. aeruginosa strain PAO1 into the rat prostate. It was found that Ps. aeruginosa produced acute and chronic infections in this mucosal tissue as determined by bacterial colonization, gross morphology, tissue damage and inflammatory markers. WT strain PAO1 and its isogenic mutant PAO-JP2, in which both the lasI and rhlI quorum-sensing signal systems have been silenced, were compared during both acute and chronic prostate infections. In acute infections, bacterial numbers and inflammatory markers were comparable between WT PA01 and PAO-JP2; however, considerably less tissue damage occurred in infections with PAO-JP2. Chronic infections with PAO-JP2 resulted in reduced bacterial colonization, tissue damage and inflammation as compared to WT PAO1 infections. Therefore, the quorum-sensing lasI and rhlI genes in Ps. aeruginosa affect acute prostate infections, but play a considerably more important role in maintaining chronic infections. We have thus developed a highly reproducible model for the study of Ps. aeruginosa virulence in the prostate.

Bacterial Adhesion to Oral Tissues: A Model for Infectious Diseases

1989 ◽  
Vol 68 (5) ◽  
pp. 750-760 ◽  
Author(s):  
R.J. Gibbons
Keyword(s):  
Conformational Changes ◽  
Oral Hygiene ◽  
Bacterial Adhesion ◽  
Periodontal Diseases ◽  
Bacterial Colonization ◽  
Human Saliva ◽  
Bacterial Attachment ◽  
Poor Oral Hygiene ◽  
Human Teeth ◽  
Mucosal Surfaces

The majority of bacteria which colonize humans display sharp host and tissue tropisms; consequently, relatively little is known about how they initiate colonization on mucosal surfaces. The mouth has a variety of features which have enabled it to serve as a useful model for the discovery of basic principles of host-parasite interactions occurring in mucosal environments. Early studies demonstrated that indigenous bacteria attach to surfaces of the mouth in a highly selective manner; attachment was often observed to correlate with colonization. These studies led to the recognition that bacterial attachment is an essential step for colonization in environments which contain surfaces exposed to a fluid flow. Bacterial adhesion has subsequently grown into a major area of infectious disease research. Many bacteria have been found to possess proteinaceous components, called "adhesins", on their surfaces which bind in a stereochemically specific manner to complementary molecules, or "receptors", on the tissue surface. Adhesins are often lectins which bind to saccharide receptors, but some adhesins are thought to bind to proteinaceous receptors. Studies of components of human saliva, which adsorb to hydroxyapatite (HA) surfaces similar to those of teeth, and promote the attachment of prominent plaque bacteria, have revealed that the acidic proline-rich proteins (PRPs) promote the attachment of several important bacteria. These include strains of Actinomyces viscosus, Bacteroides gingival is, some strains of Streptococcus mutans, and others. The salivary PRP's are a unique family of molecules. However, segments of PRPs are structurally related to collagen. This may be significant, since B. gingivalis and certain cariogenic streptococci bind to collagenous substrata, and such interactions may facilitate their invasion into gingival tissues, or into dentin or cementum, respectively. Another unexpected observation was that although A. viscosus and other bacteria bind avidly to PRPs adsorbed onto apatitic surfaces, they do not interact with PRPs in solution. PRP molecules evidently undergo a conformational change when they adsorb to HA, and adhesins of A. viscosus recognize cryptic segments which are only exposed in adsorbed molecules. This provides the bacteria with a mechanism for efficiently attaching to teeth while suspended in saliva. It also offers a molecular explanation for their sharp tropisms for human teeth. It has proven convenient to refer to such hidden receptors for bacterial adhesins as "cryptitopes" (from cryptic, meaning hidden, and topo, meaning place). The generation of cryptitopes due to conformational changes or because of enzymatic modifications appears to be involved in the colonization of several bacteria on mucosal surfaces. In addition, there is evidence which suggests that elevated levels of neuraminidases and proteases associated with poor oral hygiene and gingivitis may also generate cryptitopes which promote colonization of certain Gram-negative bacteria associated with destructive periodontal diseases. These enzymes concurrently destroy receptors required for attachment of relatively benign species such as S. mitis and S. sanguis. Thus, the elevated levels of enzymes previously reported present in crevicular fluid and saliva of individuals with poor oral hygiene appear to have the potential to modulate bacterial colonization.

Decreased Bacterial Adhesion to Surface-Treated Titanium

2005 ◽  
Vol 28 (7) ◽  
pp. 718-730 ◽  
Author(s):  
B. Del Curto ◽  
M.F. Brunella ◽  
C. Giordano ◽  
M.P. Pedeferri ◽  
V Valtulina ◽  
...  
Keyword(s):  
Titanium Oxide ◽  
Bacterial Adhesion ◽  
Bacterial Colonization ◽  
Antibacterial Properties ◽  
Bacterial Attachment ◽  
Treatment Results ◽  
Bone Implant ◽  
Crystalline Layer ◽  
Progressive Loss ◽  
Modification Treatment

Osteointegrative dental implants are widely used in implantology for their well-known excellent performance once implanted in the host. Remarkable bacterial colonization along the transgingival region may result in a progressive loss of adhesion at gum-implant interface and an increase of the bone area exposed to pathogens. This phenomenon may negatively effect the osteointegration process and cause, in the most severe cases, implant failure. The presence of bacteria at implant site affect the growth of new bone tissue and consequently, the achievement of a mechanically stable bone-implant interface, key parameters for a suitable implant osteointegration. In the present work, a novel surface treatment has been developed and optimized in order to convert the amorphous titanium oxide in a crystalline layer enriched in anatase capable of providing not only antibacterial properties but also of stimulating the precipitation of apatite when placed in simulated body fluid. The collected data have shown that the tested treatment results in a crystalline anatase-type titanium oxide layer able to provide a remarkable decrease in bacterial attachment without negatively effecting cell metabolic activity. In conclusion, the surface modification treatment analyzed in the present study might be an elegant way to reduce the risk of bacterial adhesion and increase the lifetime of the transgingival component in the osteointegrated dental implant.

scholarly journals Rethinking Pyogenic Flexor Tenosynovitis: Biofilm Formation Treated in a Cadaveric Model

2017 ◽  
Vol 09 (03) ◽  
pp. 131-138 ◽  
Author(s):  
Constantinos Ketonis ◽  
Noreen Hickock ◽  
Asif Ilyas
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Bacterial Colonization ◽  
Bacterial Load ◽  
Bacterial Attachment ◽  
Laser Scanning Microscopy ◽  
Local Antibiotics ◽  
Saline Irrigation ◽  
Pyogenic Flexor Tenosynovitis ◽  
Flexor Tenosynovitis

Introduction Pyogenic flexor tenosynovitis (PFT) of the hand remains a challenging problem that often requires surgical irrigation and parenteral or oral antibiotics. The authors hypothesize that the pathophysiology and microenvironment of PFT can be likened to that of periprosthetic joint infections (PJIs), in which bacteria thrive in a closed synovial space with limited blood supply. As such, they postulate that PFT is also facilitated by bacterial attachment and biofilm formation rendering standard treatments less effective. In this study, they evaluate infected tendons for the presence of biofilm and explore new treatment strategies. Methods Fresh human cadaveric hand tendons were harvested and divided into 0.5-cm segments. Samples were sterilized and inoculated with 1 × 104 CFU/mL green fluorescent Staphylococcus aureus (GFP-SA) for 48 hours at 37°C. After saline washing to remove plank tonic bacteria, samples were treated for 24 hours with (1) saline irrigation, (2) antibiotics (vancomycin), (3) corticosteroids, or (4) antibiotics/corticosteroid combined. Samples were visualized using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Results Following bacterial challenge, CLSM revealed heterogeneous green fluorescence representing bacterial attachment with dense biofilm formation. SEM at > 3,000X, also demonstrated bacterial colonization in grape-like clusters consisted with a thick matrix characteristic of biofilm. Bacterial load by direct colony counting decreased by 18.5% with saline irrigation alone, 42.6% with steroids, 54.4% with antibiotics, and 77.3% with antibiotics/steroids combined (p < 0.05). Conclusion Staphylococcus aureus readily formed thick biofilm on human cadaveric tendons. The addition of both local antibiotics and corticosteroids resulted in greater decreases in biofilm formation on flexor tendons than the traditional treatment of saline irrigation alone. We suggest rethinking the current treatment of PFT and recommend considering a strategy more analogous to PJI management with the adjunctive use of local antibiotics, corticosteroids, and mechanical agitation.

scholarly journals Bacterial Adhesion at Synthetic Surfaces

1999 ◽  
Vol 65 (11) ◽  
pp. 4995-5002 ◽  
Author(s):  
D. Cunliffe ◽  
C. A. Smart ◽  
C. Alexander ◽  
E. N. Vulfson
Keyword(s):  
Molecular Weight ◽  
Protein Adsorption ◽  
Bacterial Adhesion ◽  
Cell Attachment ◽  
Systematic Investigation ◽  
Bacterial Attachment ◽  
E Coli ◽  
Glass Substrates ◽  
Cell Adsorption

ABSTRACT A systematic investigation into the effect of surface chemistry on bacterial adhesion was carried out. In particular, a number of physicochemical factors important in defining the surface at the molecular level were assessed for their effect on the adhesion ofListeria monocytogenes, Salmonella typhimurium,Staphylococcus aureus, and Escherichia coli. The primary experiments involved the grafting of groups varying in hydrophilicity, hydrophobicity, chain length, and chemical functionality onto glass substrates such that the surfaces were homogeneous and densely packed with functional groups. All of the surfaces were found to be chemically well defined, and their measured surface energies varied from 15 to 41 mJ · m−2. Protein adsorption experiments were performed with3H-labelled bovine serum albumin and cytochromec prior to bacterial attachment studies. Hydrophilic uncharged surfaces showed the greatest resistance to protein adsorption; however, our studies also showed that the effectiveness of poly(ethyleneoxide) (PEO) polymers was not simply a result of its hydrophilicity and molecular weight alone. The adsorption of the two proteins approximately correlated with short-term cell adhesion, and bacterial attachment for L. monocytogenes and E. coli also correlated with the chemistry of the underlying substrate. However, for S. aureus and S. typhimurium a different pattern of attachment occurred, suggesting a dissimilar mechanism of cell attachment, although high-molecular-weight PEO was still the least-cell-adsorbing surface. The implications of this for in vivo attachment of cells suggest that hydrophilic passivating groups may be the best method for preventing cell adsorption to synthetic substrates provided they can be grafted uniformly and in sufficient density at the surface.

scholarly journals Pharmacodynamics of RWJ-54428 against Staphylococcus aureus, Streptococcus pneumoniae, and Enterococcus faecalis in a Neutropenic Mouse Thigh Infection Model

2008 ◽  
Vol 52 (1) ◽  
pp. 244-247 ◽  
Author(s):  
David C. Griffith ◽  
David Rodriguez ◽  
Erik Corcoran ◽  
Michael N. Dudley
Keyword(s):  
Staphylococcus Aureus ◽  
Streptococcus Pneumoniae ◽  
Enterococcus Faecalis ◽  
Infection Model ◽  
Antibacterial Effects ◽  
Bacteriostatic Effect ◽  
Dosing Interval ◽  
Gram Positive Bacteria ◽  
Initiation Of Therapy

ABSTRACT RWJ-54428 (also known as MC-02,479) is a new cephalosporin with promising activity against gram-positive bacteria. The pharmacodynamics (PDs) of RWJ-54428 against Staphylococcus aureus, Streptococcus pneumoniae, and Enterococcus faecalis were studied in a neutropenic mouse thigh infection model. The RWJ-54428 MICs ranged from 0.25 to 1 mg/liter. Mice with ca. 106 CFU/thigh at the initiation of therapy were treated intraperitoneally with RWJ-54428 at doses that ranged from 3 to 1,200 mg/kg of body weight/day (in 2, 3, 4, 6, or 12 divided doses) for 24 h. The maximal reductions in bacterial counts in thigh tissues at 24 h for the methicillin-resistant S. aureus, penicillin-resistant S. pneumoniae, and E. faecalis strains were −2.8, −3.8, and −1.7 log10 CFU/thigh, respectively. The percentage of a 24-h dosing interval that the unbound serum RWJ-54428 concentrations exceeded the MIC (fT > MIC) was the pharmacokinetic (PK)-PD parameter that best described the efficacy of RWJ-54428. The fT > MICs for a bacteriostatic effect (no net change in the numbers of CFU/thigh over 24 h) ranged from 14 to 20% for staphylococci and streptococci; for maximal reductions in the numbers of CFU/thigh, the fT > MICs ranged from 22 to 36% for these strains. For E. faecalis, the ranges of fT > MICs for static and maximal effects were 30 to 46% and 55 to 60%, respectively. These data show that treatment with RWJ-54428 results in marked antibacterial effects in vivo, with the PK-PD parameters for efficacy being comparable to those for the efficacy of penicillins and carbapenems active against staphylococci and pneumococci.

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