Eradication Of Preformed Biofilms Research Articles

Exploration of Antibiofilm Activities of Pediococci Strains Against Diarrheal Multi-Drug Resistant E.coli Strains

ABSTRACT The present study reports the potential of cell-free supernatant (CFS) from different probiotics for its antimicrobial and antibiofilm activity against Multi-Drug Resistant Diarrheal E. coli (MDR-DEC) strains. The CFS from Pediococcus acidilactici 100 exhibited maximum antimicrobial activity with Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of 23.33 mg/mL and 33.33 mg/mL, respectively. The CFS was able to inhibit the process of biofilm formation as well as eradication of preformed biofilms by 72–73%. A complete disintegration of aggregation of cells was observed in treated biofilms through advanced microscopic techniques. Atomic force microscopy (AFM) revealed a decrease in roughness (30%) and height of treated biofilms (2–17%). Organic acids, fatty acids, phenolic compounds and other derivatives were determined in CFS fractions using Liquid Chromatography Mass Spectrometry. The findings demonstrated the potential of CFS components from Pediococci strains against biofilms of MDR-DEC strains.

Development and Antibacterial Properties of 4-[4-(Anilinomethyl)-3-phenylpyrazol-1-yl]benzoic Acid Derivatives as Fatty Acid Biosynthesis Inhibitors.

A number of novel pyrazole derivatives have been synthesized, and several of these compounds are potent antibacterial agents with minimum inhibitory concentrations as low as 0.5 μg/mL. Human cell lines were tolerant to these lead compounds, and they showed negligible hemolytic effects at high concentrations. These bactericidal compounds are very effective against bacterial growth in both planktonic and biofilm contexts. Various techniques were applied to show the inhibition of biofilm growth and eradication of preformed biofilms by lead compounds. Potent compounds are more effective against persisters than positive controls. In vivo studies revealed that lead compounds are effective in rescuing C. elegans from bacterial infections. Several methods were applied to determine the mode of action including membrane permeability assay and SEM micrograph studies. Furthermore, CRISPRi studies led to the determination of these compounds as fatty acid biosynthesis (FAB) inhibitors.

Antibacterial and Antibiofilm Properties of the Alexidine Dihydrochloride (MMV396785) against Acinetobacter baumannii.

Antibiotic-resistant Acinetobacter baumannii infections among patients in hospital settings are rising at an alarming rate. The World Health Organization has designated carbapenem-resistant A. baumannii as a priority pathogen for drug discovery. Based on the open drug discovery approach, we screened 400 compounds provided as a Pandemic Response Box by MMV and DNDi to identify compounds with antibacterial and antibiofilm activity against two A. baumannii reference strains using a highly robust resazurin assay. In vitro screening identified thirty compounds with MIC ≤ 50μM having growth inhibitory properties against the planktonic state. Five compounds, with MMV IDs MMV396785, MMV1578568, MMV1578574, MMV1578564, and MMV1579850, were able to reduce metabolically active cells in the biofilm state. Of these five compounds, MMV396785 showed potential antibacterial and antibiofilm activity with MIC, MBIC, and MBEC of 3.125 μM, 12.5, and 25-100 µM against tested A. baumannii strains, respectively, showing biofilm formation inhibition by 93% and eradication of pre-formed biofilms by 60-77.4%. In addition, MMV396785 showed a drastic reduction in the surface area and thickness of biofilms. Further investigations at the molecular level by qRT-PCR revealed the downregulation of biofilm-associated genes when exposed to 50 µM MMV396785 in all tested strains. This study identified the novel compound MMV396785 as showing potential in vitro antibacterial and antibiofilm efficacy against A. baumannii.

Gold-, silver- and magnesium-doped zinc oxide nanoparticles prevents the formation of and eradicates bacterial biofilms.

Aim: This work aimed to synthesize magnesium-doped zinc oxide, silver and gold nanoparticles (Nps) and to evaluate their potential to prevent and eradicate Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Acinetobacter baumannii and Pseudomonas aeruginosa biofilms. Materials & methods: The Nps were synthesized by precipitation and metallic reduction techniques. Physicochemical and biological characterization of Nps was performed. Results: All the Nps tested were able to inhibit the formation of E. coli, P. mirabilis, S. aureus and A. baumannii biofilms. The effects on the eradication of preformed biofilms were variable, although all the Nps tested were able to eradicate A. baumannii biofilms. Conclusion: The observed effects make the Nps suitable for coating surfaces and/or antibiotic carriers with medical interest.

Tailored anti-biofilm activity – Liposomal delivery for mimic of small antimicrobial peptide

The eradication of bacteria embedded in biofilms is among the most challenging obstacles in the management of chronic wounds. These biofilms are found in most chronic wounds; moreover, the biofilm-embedded bacteria are considerably less susceptible to conventional antimicrobial treatment than the planktonic bacteria. Antimicrobial peptides and their mimics are considered attractive candidates in the pursuit of novel therapeutic options for the treatment of chronic wounds and general bacterial eradication. However, some limitations linked to these membrane-active antimicrobials are making their clinical use challenging. Novel innovative delivery systems addressing these limitations represent a smart solution. We hypothesized that incorporation of a novel synthetic mimic of an antimicrobial peptide in liposomes could improve its anti-biofilm effect as well as the anti-inflammatory activity. The small synthetic mimic of an antimicrobial peptide, 7e-SMAMP, was incorporated into liposomes (~280 nm) tailored for skin wounds and evaluated for its potential activity against both biofilm formation and eradication of pre-formed biofilms. The 7e-SMAMP-liposomes significantly lowered inflammatory response in murine macrophages (~30 % reduction) without affecting the viability of macrophages or keratinocytes. Importantly, the 7e-SMAMP-liposomes completely eradicated biofilms produced by Staphylococcus aureus and Escherichia coli above concentrations of 6.25 μg/mL, whereas in Pseudomonas aeruginosa the eradication reached 75 % at the same concentration. Incorporation of 7e-SMAMP in liposomes improved both the inhibition of biofilm formation as well as biofilm eradication in vitro, as compared to non-formulated antimicrobial, therefore confirming its potential as a novel therapeutic option for bacteria-infected chronic wounds.

Lactoferrin modulates the biofilm formation and bap gene expression of methicillin-resistant Staphylococcus epidermidis.

Lactoferrin is an innate glycoprotein with broad antibacterial and antibiofilm properties. The autonomous antibiofilm activity of lactoferrin against Gram-positive bacteria is postulated to involve the cell wall and biofilm components. Thus, the prevention of biomass formation and eradication of preformed biofilms by lactoferrin was investigated using a methicillin-resistant Staphylococcus epidermidis (MRSE) strain. Additionally, the ability of lactoferrin to modulate the expression of the biofilm-associated protein gene (bap) was studied. The bap gene regulates the production of biofilm-associated proteins responsible for bacterial adhesion and aggregation. In the in vitro biofilm assays, lactoferrin prevented biofilm formation and eradicated established biofilms for up to 24 and 72 h, respectively. Extensive eradication of MRSE biofilm biomass was accompanied by the significant upregulation of bap gene expression. These data suggest the interaction of lactoferrin with the biofilm components and cell wall of MRSE, including the biofilm-associated protein.

Polymyxin B in Combination with Glycerol Monolaurate Exerts Synergistic Killing against Gram-Negative Pathogens.

The rapid emergence and spread of multidrug-resistant (MDR) bacterial pathogens pose a serious danger to worldwide human health, and resistance to last-resort drugs, such as polymyxins, is being increasingly detected in MDR Gram-negative pathogens. There is an urgent need to find and optimize combination therapies as an alternative therapeutic strategy, with a dry pipeline in novel antibiotic research and development. We found a monoester formed from the combination of lauric acid and glycerol, glycerol monolaurate (GML), possessing prominent antibacterial and anti-inflammatory activity. However, it is still unclear whether GML in combination could increase antimicrobial activity. Here, we reported that polymyxin B (PMNB) combined with GML exhibited a synergistic antimicrobial impact on Gram-negative strains in vitro, including clinical MDR isolates. This synergistic antimicrobial activity correlated with the destruction of bacterial cell structures, eradication of preformed biofilms, and increased reactive oxygen species (ROS) accumulation. We also showed that PMNB synergized with GML effectively eliminated pathogens from bacterial pneumonia caused by Klebsiella pneumoniae to rescue mice. Our research demonstrated that the PMNB and GML combination induced synergistic antimicrobial activity for Gram-negative pathogens in vitro and in vivo. These findings are of great importance for treating bacterial infections and managing the spread of infectious diseases.

Activity of Biodegradable Polymeric Nanosponges against Dual-Species Bacterial Biofilms.

Infections caused by multidrug-resistant (MDR) bacteria present an emerging global health crisis, and the threat is intensified by the involvement of biofilms. Some biofilm infections involve more than one species; this can further challenge treatment using traditional antibiotics. Nanomaterials are being developed as alternative therapeutics to traditional antibiotics; here we report biodegradable polymer-stabilized oil-in-water nanosponges (BNS) and show their activity against dual-species bacterial biofilms. The described engineered nanosponges demonstrated broad-spectrum antimicrobial activity through prevention of dual-species biofilm formation as well as eradication of preformed biofilms. The BNS showed no toxicity against mammalian cells. Together, these data highlight the therapeutic potential of this platform.

Naringin inhibits the biofilms of metallo-β-lactamases (MβLs) producing Pseudomonas species isolated from camel meat.

Food producing animals harbouring bacteria carrying drug resistance genes especially the metallo-beta-lactamase (MBL) pose high risk for the human population. In addition, formation of biofilm by these drug resistant pathogens represents major threat to food safety and public health. In this study, metallo-β-lactamases (MβLs) producing Pseudomonas spp. from camel meat were isolated and assessed for their biofilm formation. Further, in vitro and in silico studies were performed to study the effect of flavone naringin on biofilm formation against isolated Pseudomonas spp. A total of 55% isolates were found to produce metallo-β-lactamase enzyme. Naringin mitigated biofilm formation of Pseudomonas isolates up to 57%. Disturbed biofilm architecture and reduced the colonization of bacteria on glass was observed under scanning electron microscope (SEM) and confocal laser scanning microscope (CLSM). The biofilm related traits such as exopolysaccharides (EPS) and alginate production was also reduced remarkably in the presence of naringin. Eradication of preformed biofilms (32–60%) was also observed at the respective 0.50 × MICs. Molecular docking revealed that naringin showed strong affinity towards docked proteins with binding energy ranging from −8.6 to −8.8 kcal mol−1. Presence of metallo-β-lactamase producers indicates that camel meat could be possible reservoir of drug-resistant Pseudomonas species of clinical importance. Naringin was successful in inhibiting biofilm formation as well as eradicating the preformed biofilms and demonstrated strong binding affinity towards biofilm associated protein. Thus, it is envisaged that naringin could be exploited as food preservative especially against the biofilm forming food-borne Pseudomonas species and is a promising prospect for the treatment of biofilm based infections.

Utilizing nanoparticles for improving anti-biofilm effects of azithromycin: A head-to-head comparison of modified hyaluronic acid nanogels and coated poly (lactic-co-glycolic acid) nanoparticles

HypothesisThe widespread resistance of bacteria to traditional antibiotic treatments has expedited the search for novel therapies against these pathogens. The hypothesis of this work is that two distinctively different polymeric delivery systems, specifically D-α-tocopherol polyethylene glycol 1000 succinate (TPGS)-poly(lactic-co-glycolic acid) (PLGA) nanoparticles and octenyl succinic anhydride-modified low molecular weight hyaluronic acid (OSA-HA) nanogels may be used to substantially improve the properties of azithromycin, allowing its use for effective treatment of Pseudomonas aeruginosa biofilm infections. ExperimentsAzithromycin was encapsulated in both delivery systems and the physicochemical properties of the loaded delivery systems, including size, surface charge and drug loading were evaluated. Additionally, particle interaction with a mucin layer, penetration into a bacterial biofilm, prevention of biofilm formation and eradication of pre-formed biofilms, the influence on production of virulence factors and bacterial motility as well as cytotoxicity towards hepatocytes and lung epithelial cells were compared head-to-head. FindingsThe TPGS-PLGA nanoparticles noticeably improved the antimicrobial activity and the biofilm prevention activity of azithromycin whereas the OSA-HA nanogels showed reduced mucin interactions together with improved reduction of pre-formed biofilms and maintained the low eukaryotic cell cytotoxicity of azithromycin.

Peptide dendrimers G3KL and TNS18 inhibit Pseudomonas aeruginosa biofilms.

Herein we report that peptide dendrimers G3KL and TNS18, which were recently reported to control multidrug-resistant bacteria such as Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii, strongly inhibit biofilm formation by P. aeruginosa PA14 below their minimum inhibitory concentration (MIC) value, under which conditions they also strongly affect swarming motility. Eradication of preformed biofilms, however, required concentrations above the MIC values. Scanning electron microscopy observation and confocal laser scanning micrographs showed that peptide dendrimers can destroy the biofilm morphological structure and thickness in a dose-dependent manner, even make the biofilm dispersed completely. Membrane potential analysis indicated that planktonic cells treated with peptide dendrimers presented an increase in fluorescence intensity, suggesting that cytoplasmic membrane could be the target of G3KL and TNS18 similarly to polymyxin B. RNA-seq analysis showed that the expressions of genes in the arnBCADTEF operon-regulating lipid A modification resulting in resistance to AMPs are differentially affected between these three compounds, suggesting that each compound targets the cell membrane but in different manner. Potent activity on planktonic cells and biofilms of P. aeruginosa suggests that peptide dendrimers G3KL and TNS18 are promising candidates of clinical development for treating infections.

Utilization of supercritical carbon dioxide in fabrication of cellulose acetate films with anti-biofilm effects against Pseudomonas aeruginosa and Staphylococcus aureus

This study discusses utilization of supercritical carbon dioxide for impregnation of cellulose acetate films with thymol in order to produce material with anti-biofilm activity against Pseudomonas aeruginosa and Staphylococcus aureus. Analysis of anti-biofilm activity of cellulose acetate beads impregnated with thymol suggested that optimal thymol loading was in the range from 26% to 30% for efficient reduction of biofilm formation and eradication of pre-formed biofilms. Polymer films were fabricated by the solvent casting method from polymer solutions of different contents, and loaded with thymol using supercritical carbon dioxide at 15.5 MPa and 35 °C. The film containing 30% of thymol (F1 30%) exhibited substantial anti-adhesion properties inhibiting biofilm formation on its surface and considerably reduced formation of biofilms on the surrounding surfaces (up to 80%) by all tested strains including antibiotic resistant P. aeruginosa DM50 and methicillin-resistant S. aureus.

Broad‐spectrum inhibitory effect of green synthesised silver nanoparticles from Withania somnifera (L.) on microbial growth, biofilm and respiration: a putative mechanistic approach

Multi-drug resistance in pathogenic bacteria has created immense clinical problem globally. To address these, there is need to develop new therapeutic strategies to combat bacterial infections. Silver nanoparticles (AgNPs) might prove to be next generation nano-antibiotics. However, improved efficacy and broad-spectrum activity is still needed to be evaluated and understood. The authors have synthesised AgNPs from Withania somnifera (WS) by green process and characterised. The effect of WS-AgNPs on growth kinetics, biofilm inhibition as well as eradication of preformed biofilms on both gram-positive and gram-negative pathogenic bacteria was evaluated. The authors have demonstrated the inhibitory effect on bacterial respiration and disruption of membrane permeability and integrity. It was found that WS-AgNPs inhibited growth of pathogenic bacteria even at 16 µg/ml. At sub-minimum inhibitory concentration concentration, there was approximately 50% inhibition in biofilm formation which was further validated by light and electron microscopy. WS-AgNPs also eradicated the performed biofilms by varying levels at elevated concentration. The bacterial respiration was also significantly inhibited. Interaction of WS-AgNPs with test pathogen caused the disruption of cell membrane leading to leakage of cellular content. The production of intracellular reactive oxygen species reveals that WS-AgNPs exerted oxidative stress inside bacterial cell causing microbial growth inhibition and disrupting cellular functions.

Removing Legionella pneumophila and biofilms from water supply systems using plant essential oils

Antibacterial and antibiofilm activities of sage (Salvia officinalis), thyme (Thymus vulgaris), peppermint (Mentha x piperita) and lemon (Citrus limonum) essential oils (EOs), supplied as commercial preparations, were investigated on Legionella pneumophila. Results showed that all EOs had antibacterial activity. Lemon oil demonstrated the best activity, with minimum inhibition concentration (MIC) and minimum bactericidal concentration values detected as 0.83–1.25 and 1.25–3.75 μL/mL, respectively. Antibiofilm activities of EOs were studied in two ways: (1) inhibition of biofilm formation at subMIC concentrations; and (2) eradication of preformed biofilms at MIC and subMIC concentrations. The highest biofilm inhibition values were 63.3, 45.5, 35.6 and 23%, and the highest eradication values were 34.2, 16.7, 39.4 and 31.9% for lemon oil, peppermint oil, sage oil and thyme oil, respectively. Lemon oil was detected to have the highest antibiofilm activity, as with antibacterial activity. The minimum antibiofilm activity was found for thyme oil. For all the EOs, the inhibition of biofilm formation was higher than the eradication of preformed biofilms at the same concentrations. The results suggested that EOs of lemon, peppermint, sage and thyme had the potential for being used in the removal of L. pneumophila from potable water systems, due to their antibacterial and antibiofilm activities.