scholarly journals Antibiotic-Loaded Synthetic Calcium Sulfate Beads for Prevention of Bacterial Colonization and Biofilm Formation in Periprosthetic Infections

2014 ◽  
Vol 59 (1) ◽  
pp. 111-120 ◽  
Author(s):  
R. P. Howlin ◽  
M. J. Brayford ◽  
J. S. Webb ◽  
J. J. Cooper ◽  
S. S. Aiken ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Calcium Sulfate ◽  
Periprosthetic Infection ◽  
Bacterial Colonization ◽  
Viable Cell ◽  
Surgical Removal ◽  
Content Type ◽  
Cell Counts ◽  
Antibiotic Release ◽  
Prosthesis Material

ABSTRACTPeriprosthetic infection (PI) causes significant morbidity and mortality after fixation and joint arthroplasty and has been extensively linked to the formation of bacterial biofilms. Poly(methyl methacrylate) (PMMA), as a cement or as beads, is commonly used for antibiotic release to the site of infection but displays variable elution kinetics and also represents a potential nidus for infection, therefore requiring surgical removal once antibiotics have eluted. Absorbable cements have shown improved elution of a wider range of antibiotics and, crucially, complete biodegradation, but limited data exist as to their antimicrobial and antibiofilm efficacy. Synthetic calcium sulfate beads loaded with tobramycin, vancomycin, or vancomycin-tobramycin dual treatment (in a 1:0.24 [wt/wt] ratio) were assessed for their abilities to eradicate planktonic methicillin-resistantStaphylococcus aureus(MRSA) andStaphylococcus epidermidisrelative to that of PMMA beads. The ability of the calcium sulfate beads to prevent biofilm formation over multiple days and to eradicate preformed biofilms was studied using a combination of viable cell counts, confocal microscopy, and scanning electron microscopy of the bead surface. Biofilm bacteria displayed a greater tolerance to the antibiotics than their planktonic counterparts. Antibiotic-loaded beads were able to kill planktonic cultures of 106CFU/ml, prevent bacterial colonization, and significantly reduce biofilm formation over multiple days. However, established biofilms were harder to eradicate. These data further demonstrate the difficulty in clearing established biofilms; therefore, early preventive measures are key to reducing the risk of PI. Synthetic calcium sulfate loaded with antibiotics has the potential to reduce or eliminate biofilm formation on adjacent periprosthetic tissue and prosthesis material and, thus, to reduce the rates of periprosthetic infection.

scholarly journals The Urinary Antibiotic 5-Nitro-8-Hydroxyquinoline (Nitroxoline) Reduces the Formation and Induces the Dispersal of Pseudomonas aeruginosa Biofilms by Chelation of Iron and Zinc

2012 ◽  
Vol 56 (11) ◽  
pp. 6021-6025 ◽  
Author(s):  
A. Sobke ◽  
M. Klinger ◽  
B. Hermann ◽  
S. Sachse ◽  
S. Nietzsche ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Broad Spectrum ◽  
Viable Cell ◽  
Antibiofilm Activity ◽  
Content Type ◽  
Cell Counts ◽  
Iron And Zinc ◽  
Chelation Of Iron

ABSTRACTSince cations have been reported as essential regulators of biofilm, we investigated the potential of the broad-spectrum antimicrobial and cation-chelator nitroxoline as an antibiofilm agent. Biofilm mass synthesis was reduced by up to 80% at sub-MIC nitroxoline concentrations inPseudomonas aeruginosa, and structures formed were reticulate rather than compact. In preformed biofilms, viable cell counts were reduced by 4 logs at therapeutic concentrations. Complexation of iron and zinc was demonstrated to underlie nitroxoline's potent antibiofilm activity.

scholarly journals Bactericidal Activity of ACH-702 against Nondividing and Biofilm Staphylococci

2012 ◽  
Vol 56 (7) ◽  
pp. 3812-3818 ◽  
Author(s):  
Steven D. Podos ◽  
Jane A. Thanassi ◽  
Melissa Leggio ◽  
Michael J. Pucci
Keyword(s):  
Protein Synthesis ◽  
Stationary Phase ◽  
Bactericidal Activity ◽  
Bacterial Infections ◽  
De Novo ◽  
Viable Cell ◽  
Bacterial Cells ◽  
Protein Synthesis Inhibitors ◽  
Content Type ◽  
Cell Counts

ABSTRACTMany bacterial infections involve slow or nondividing bacterial growth states and localized high cell densities. Antibiotics with demonstrated bactericidal activity rarely remain bactericidal at therapeutic concentrations under these conditions. The isothiazoloquinolone (ITQ) ACH-702 is a potent, bactericidal compound with activity against many antibiotic-resistant pathogens, including methicillin-resistantStaphylococcus aureus(MRSA). We evaluated its bactericidal activity under conditions where bacterial cells were not dividing and/or had slowed their growth. AgainstS. aureuscultures in stationary phase, ACH-702 showed concentration-dependent bactericidal activity and achieved a 3-log-unit reduction in viable cell counts within 6 h of treatment at ≥16× MIC values; in comparison, the bactericidal quinolone moxifloxacin and the additional comparator compounds vancomycin, linezolid, and rifampin at 16× to 32× MICs showed little or no bactericidal activity against stationary-phase cells. ACH-702 at 32× MIC retained bactericidal activity against stationary-phaseS. aureusacross a range of inoculum densities. ACH-702 did not kill cold-arrested cells yet remained bactericidal against cells arrested by protein synthesis inhibitors, suggesting that its bactericidal activity against nondividing cells requires active metabolism but notde novoprotein synthesis. ACH-702 also showed a degree of bactericidal activity at 16× MIC againstS. epidermidisbiofilm cells that was superior to that of moxifloxacin, rifampin, and vancomycin. The bactericidal activity of ACH-702 against stationary-phase staphylococci and biofilms suggests potential clinical utility in infections containing cells in these physiological states.

Antibacterial and antibiofilm effects of iron chelators against Prevotella intermedia

2013 ◽  
Vol 62 (9) ◽  
pp. 1307-1316 ◽  
Author(s):  
Ji-Hoi Moon ◽  
Cheul Kim ◽  
Hee-Su Lee ◽  
Sung-Woon Kim ◽  
Jin-Yong Lee
Keyword(s):  
Bacterial Growth ◽  
Biofilm Formation ◽  
Therapeutic Agent ◽  
Viable Cell ◽  
Iron Chelators ◽  
Prevotella Intermedia ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Cell Counts ◽  
Strong Antimicrobial Activity

Prevotella intermedia, a major periodontopathogen, has been shown to be resistant to many antibiotics. In the present study, we examined the effect of the FDA-approved iron chelators deferoxamine (DFO) and deferasirox (DFRA) against planktonic and biofilm cells of P. intermedia in order to evaluate the possibility of using these iron chelators as alternative control agents against P. intermedia. DFRA showed strong antimicrobial activity (MIC and MBC values of 0.16 mg ml−1) against planktonic P. intermedia. At subMICs, DFRA partially inhibited the bacterial growth and considerably prolonged the bacterial doubling time. DFO was unable to completely inhibit the bacterial growth in the concentration range tested and was not bactericidal. Crystal violet binding assay for the assessment of biofilm formation by P. intermedia showed that DFRA significantly decreased the biofilm-forming activity as well as the biofilm formation, while DFO was less effective. DFRA was chosen for further study. In the ATP-bioluminescent assay, which reflects viable cell counts, subMICs of DFRA significantly decreased the bioactivity of biofilms in a concentration-dependent manner. Under the scanning electron microscope, P. intermedia cells in DFRA-treated biofilm were significantly elongated compared to those in untreated biofilm. Further experiments are necessary to show that iron chelators may be used as a therapeutic agent for periodontal disease.

scholarly journals Novel Two-Component System ofStreptococcus sanguinisAffecting Functions Associated with Viability in Saliva and Biofilm Formation

2018 ◽  
Vol 86 (4) ◽  
Author(s):  
Tarsila M. Camargo ◽  
Rafael N. Stipp ◽  
Lívia A. Alves ◽  
Erika N. Harth-Chu ◽  
José F. Höfling ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Ex Vivo ◽  
Opportunistic Pathogen ◽  
Dna Looping ◽  
Extracellular Dna ◽  
Two Component System ◽  
Component System ◽  
Content Type ◽  
Cell Counts ◽  
Two Component

ABSTRACTStreptococcus sanguinisis a pioneer species of teeth and a common opportunistic pathogen of infective endocarditis. In this study, we identified a two-component system,S. sanguinisSptRS (SptRSSs), affectingS. sanguinissurvival in saliva and biofilm formation. Isogenic mutants ofsptRSs(SKsptR) andsptSSs(SKsptS) showed reduced cell counts inex vivoassays of viability in saliva compared to those of parent strain SK36 and complemented mutants. Reduced counts of the mutants in saliva were associated with reduced growth rates in nutrient-poor medium (RPMI) and increased susceptibility to the deposition of C3b and the membrane attach complex (MAC) of the complement system, a defense component of saliva and serum. Conversely,sptRSsandsptSSsmutants showed increased biofilm formation associated with higher levels of production of H2O2and extracellular DNA. Reverse transcription-quantitative PCR (RT-qPCR) comparisons of strains indicated a global role of SptRSSsin repressing genes for H2O2production (2.5- to 15-fold upregulation ofspxB,spxR,vicR,tpk, andackAinsptRSsandsptSSsmutants), biofilm formation, and/or evasion of host immunity (2.1- to 11.4-fold upregulation ofsrtA,pcsB,cwdP,iga, andnt5e). Compatible with the homology of SptRSswith AraC-type regulators, duplicate to multiple conserved repeats were identified in 1,000-bp regulatory regions of downstream genes, suggesting that SptRSsregulates transcription by DNA looping. Significant transcriptional changes in the regulatory genesvicR,spxR,comE,comX, andmecAin thesptRSsandsptSSsmutants further indicated that SptRSSsis part of a regulatory network that coordinates cell wall homeostasis, H2O2production, and competence. This study reveals that SptRSSsis involved in the regulation of crucial functions forS. sanguinispersistence in the oral cavity.

scholarly journals Subinhibitory Concentrations of LFF571 Reduce Toxin Production by Clostridium difficile

2014 ◽  
Vol 59 (2) ◽  
pp. 1252-1257 ◽  
Author(s):  
Meena Sachdeva ◽  
Jennifer A. Leeds
Keyword(s):  
Clostridium Difficile ◽  
Elongation Factor ◽  
Viable Cell ◽  
Toxin Production ◽  
Synthesis Inhibitor ◽  
Content Type ◽  
Cell Counts ◽  
A Cell ◽  
Subinhibitory Concentrations ◽  
Bacterial Protein Synthesis

ABSTRACTLFF571 is a novel semisynthetic thiopeptide antibacterial that is undergoing investigation for safety and efficacy in patients with moderateClostridium difficileinfections. LFF571 inhibits bacterial protein synthesis by interacting with elongation factor Tu (EF-Tu) and interrupting complex formation between EF-Tu and aminoacyl-tRNA. Given this mechanism of action, we hypothesized that concentrations of LFF571 below those necessary to inhibit bacterial growth would reduce steady-state toxin levels inC. difficilecultures. We investigatedC. difficilegrowth and toxin A and B levels in the presence of LFF571, fidaxomicin, vancomycin, and metronidazole. LFF571 led to strain-dependent effects on toxin production, including decreased toxin levels after treatment with subinhibitory concentrations, and more rapid declines in toxin production than in inhibition of colony formation. Fidaxomicin, which is an RNA synthesis inhibitor, conferred a similar pattern to LFF571 with respect to toxin levels versus viable cell counts. The incubation of two toxigenicC. difficilestrains with subinhibitory concentrations of vancomycin, a cell wall synthesis inhibitor, increased toxin levels in the supernatant over those of untreated cultures. A similar phenomenon was observed with one metronidazole-treated strain ofC. difficile. These studies indicate that LFF571 and fidaxomicin generally result in decreasedC. difficiletoxin levels in culture supernatants, whereas treatment of some strains with vancomycin or metronidazole had the potential to increase toxin levels. Although the relevance of these findings remains to be studied in patients, reducing toxin levels with sub-growth-inhibitory concentrations of an antibiotic is hypothesized to be beneficial in alleviating symptoms.

scholarly journals Temperature Effect on Listeria Monocytogenes Planktonic Growth and Biofilm-Forming Ability

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Joana Catarina Andrade ◽  
◽  
Rita Bernardo ◽  
António Salvador Barreto ◽  
Telmo Nunes ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Temperature Effect ◽  
Optical Density ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Viable Cell ◽  
Crystal Violet Staining ◽  
Cell Counts ◽  
Viable Cells ◽  
Density Results

Listeria Monocytogenes is an important foodborne pathogen with the capacity to grow at low temperatures and the ability to form biofilms. These features are particularly significant to food business operators producing readyto-eat foods with a long refrigerated shelf-life not undergoing any listericidal treatment before consumption. Objectives: This work aims to assess the temperature effect on L. monocytogenes growth in planktonic suspension and in mono-species biofilms. Methods and results: Isothermal planktonic growth at 12o C and 37o C was assayed using viable cell counts and optical density measurements that revealed a strong positive correlation, confirming the reliability of combining both methods to estimate L. monocytogenes concentration. Experimental data were then fitted to Baranyi and Roberts primary predictive model and the estimated growth parameters confirmed that μmax at 37o C (0.375 ± 0.072 log Cfu/ ml/h) was higher than at 120 C (0.054 ± 0.001 log Cfu/ml/h), with identical L. monocytogenes final concentrations which emphasizes its ability to grow at refrigerated temperatures. Experimental results from the isothermal growth assay and ComBase Predictor growth model were similar, with slightly higher estimated μmax (37o C: 0.480 log Cfu/ml/h; 12o C: 0.068 log Cfu/ml/h) in the predictor growth model. The studied strains demonstrated biofilm-forming ability at 12o C, 20o C and 300 C after 5 days of growth. No significant differences in biofilm formation at different temperatures were detected considering viable cell counts values, but when using crystal violet staining optical density results significant differences were found, with the highest formation occurring at 30ºC. A positive strong correlation was found between viable cell counts and crystal violet staining optical density results. In fact, both methods complement each other, because while viable cell counts measures viable cells, crystal violet staining optical density considers total biomass (viable and non-viable cells and extracellular matrix components). Nevertheless, in this work all L. monocytogenes strains revealed to be weak biofilm producers. Conclusion: Overall, this studys results contribute with important initial information on L. monocytogenes growth and biofilm formation to further assist predictive growth modeling in food matrices and environments, also enabling subsequent quantitative microbial risk assessment, to improve pathogen’s control.

scholarly journals A Multiprotein Complex Anchors Adhesive Holdfast at the Outer Membrane ofCaulobacter crescentus

2019 ◽  
Vol 201 (18) ◽  
Author(s):  
Nina I. Sulkowski ◽  
Gail G. Hardy ◽  
Yves V. Brun ◽  
Tanmay A. M. Bharat
Keyword(s):  
Cell Surface ◽  
Outer Membrane ◽  
Biofilm Formation ◽  
Caulobacter Crescentus ◽  
Cell Envelope ◽  
Bacterial Colonization ◽  
Bacterial Species ◽  
Specific Cell ◽  
Content Type ◽  
In Cells

ABSTRACTAdhesion allows microbes to colonize surfaces and is the first stage in biofilm formation. Stable attachment of the freshwater alphaproteobacteriumCaulobacter crescentusto surfaces requires an adhesive polysaccharide called holdfast, which is synthesized at a specific cell pole and ultimately found at the tip of cylindrical extensions of the cell envelope called stalks. Secretion and anchoring of holdfast to the cell surface are governed by proteins HfsDAB and HfaABD, respectively. The arrangement and organization of these proteins with respect to each other and the cell envelope, and the mechanism by which the holdfast is anchored on cells, are unknown. In this study, we have imaged a series ofC. crescentusmutants using electron cryotomography, revealing the architecture and arrangement of the molecular machinery involved in holdfast anchoring in cells. We found that the holdfast is anchored to cells by a defined complex made up of the HfaABD proteins and that the HfsDAB secretion proteins are essential for proper assembly and localization of the HfaABD anchor. Subtomogram averaging of cell stalk tips showed that the HfaABD complex spans the outer membrane. The anchor protein HfaB is the major component of the anchor complex located on the periplasmic side of the outer membrane, while HfaA and HfaD are located on the cell surface. HfaB is the critical component of the complex, without which no HfaABD complex was observed in cells. These results allow us to propose a working model of holdfast anchoring, laying the groundwork for further structural and cell biological investigations.IMPORTANCEAdhesion and biofilm formation are fundamental processes that accompany bacterial colonization of surfaces, which are of critical importance in many infections.Caulobacter crescentusbiofilm formation proceeds via irreversible adhesion mediated by a polar polysaccharide called holdfast. Mechanistic and structural details of how the holdfast is secreted and anchored on cells are still lacking. Here, we have assigned the location and described the arrangement of the holdfast anchor complex. This work increases our knowledge of the relatively underexplored field of polysaccharide-mediated adhesion by identifying structural elements that anchor polysaccharides to the cell envelope, which is important in a variety of bacterial species.

scholarly journals Biocide Exposure Induces Changes in Susceptibility, Pathogenicity, and Biofilm Formation in Uropathogenic Escherichia coli

2019 ◽  
Vol 63 (3) ◽  
Author(s):  
E. L. Henly ◽  
J. A. R. Dowling ◽  
J. B. Maingay ◽  
M. M. Lacey ◽  
T. J. Smith ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Silver Nitrate ◽  
Biofilm Formation ◽  
Galleria Mellonella ◽  
Bacterial Colonization ◽  
Fibroblast Cell ◽  
Cross Resistance ◽  
Coating Agent ◽  
Content Type

ABSTRACT Uropathogenic Escherichia coli (UPEC) is a frequent cause of catheter-associated urinary tract infection (CAUTI). Biocides have been incorporated into catheter coatings to inhibit bacterial colonization while, ideally, exhibiting low cytotoxicity and mitigating the selection of resistant bacterial populations. We compared the effects of long-term biocide exposure on susceptibility, biofilm formation, and relative pathogenicity in eight UPEC isolates. MICs, minimum bactericidal concentrations (MBCs), minimum biofilm eradication concentrations (MBECs), and antibiotic susceptibilities were determined before and after long-term exposure to triclosan, polyhexamethylene biguanide (PHMB), benzalkonium chloride (BAC), and silver nitrate. Biofilm formation was quantified using a crystal violet assay, and relative pathogenicity was assessed via a Galleria mellonella waxworm model. Cytotoxicity and the resulting biocompatibility index values were determined by use of an L929 murine fibroblast cell line. Biocide exposure resulted in multiple decreases in biocide susceptibility in planktonic and biofilm-associated UPEC. Triclosan exposure induced the largest frequency and magnitude of susceptibility decreases at the MIC, MBC, and MBEC, which correlated with an increase in biofilm biomass in all isolates. Induction of antibiotic cross-resistance occurred in 6/84 possible combinations of bacteria, biocide, and antibiotic. Relative pathogenicity significantly decreased after triclosan exposure (5/8 isolates), increased after silver nitrate exposure (2/8 isolates), and varied between isolates for PHMB and BAC. The biocompatibility index ranked the antiseptic potential as PHMB > triclosan > BAC > silver nitrate. Biocide exposure in UPEC may lead to reductions in biocide and antibiotic susceptibility, changes in biofilm formation, and alterations in relative pathogenicity. These data indicate the multiple consequences of biocide adaptation that should be considered when selecting an anti-infective catheter-coating agent.

scholarly journals The Acinetobacter baumannii Biofilm-Associated Protein Plays a Role in Adherence to Human Epithelial Cells

2011 ◽  
Vol 80 (1) ◽  
pp. 228-233 ◽  
Author(s):  
Kari A. Brossard ◽  
Anthony A. Campagnari
Keyword(s):  
Epithelial Cells ◽  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Bacterial Colonization ◽  
Water Channel ◽  
Cell Surface Hydrophobicity ◽  
Health Care Facilities ◽  
Content Type ◽  
Abiotic Surfaces ◽  
Normal Human

ABSTRACTAcinetobacter baumanniiis a significant source of nosocomial infections worldwide. This bacterium has the ability to survive and persist on multiple abiotic surfaces in health care facilities, and once a focus has been established, this opportunistic pathogen is difficult to eradicate. This paper demonstrates that theA. baumanniibiofilm-associated protein (Bap) is necessary for mature biofilm formation on medically relevant surfaces, including polypropylene, polystyrene, and titanium. Scanning electron microscopy analyses of biofilms show that Bap is required for three-dimensional tower structure and water channel formation. In conjunction with persistence on abiotic surfaces, adherence to eukaryotic cells is an important step in bacterial colonization resulting in infection of the host. We have described Bap as the surface structure involved in adherence ofA. baumanniito both normal human bronchial epithelial cells and normal human neonatal keratinocytes. However, Bap is not involved in internalization of the bacterium in these two cell lines. Furthermore, this study shows that the presence of Bap increases the bacterial cell surface hydrophobicity. The results of this study are pertinent, as the data lead to a better understanding of the role of Bap in biofilm formation on medical surfaces and in colonization of the host.

scholarly journals Assessing toxicity of titanium dioxide (TiO2) nanoparticles on Pseudomonas species biofilms

2021 ◽  
Author(s):  
Yevheniya Chabanyuk
Keyword(s):  
Titanium Dioxide ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Titanium Dioxide Nanoparticles ◽  
Viable Cell ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Long Term Effects ◽  
Software Analysis ◽  
Cell Counts

Biofilms are essential to the aquatic environment. Recent advances in technology resulted in increased use of nanomaterials (such as titanium dioxide nanoparticles) and their release into aquatic environments with unknown long-term effects. Potential toxicity of titanium dioxide, known for its photocatalytic properties, on Pseudomonas aeruginosa (PAO1-gfp) and Pseudomonas sp. (CT07-gfp) biofilm formation and proliferation was assessed using flowcells, confocal laser scanning microscopy (CLSM), and total and viable cell release into effluent under different titanium dioxide concentrations (100 ppm, 10 ppm and 1 ppm). COMSTAT software analysis was used to obtain quantitative morphological biofilm data. Results showed that titanium dioxide had a concentration and media-dependent effect on biofilm formation, growth, proliferation and viability. Viable effluent cell counts remained within the same order of magnitude. Biofilm recovery was evident within 24-48 hours after exposure. At environmentally relevant concentration (1 ppm), there was no effect on formation, proliferation or growth of the biofilm.

Sign in / Sign up

Export Citation Format

Share Document