Potent Antibiofilm Agent Research Articles

Evaluation of local Origanum vulgare aqueous extract for eradication of biofilm production bacteria

Origanum vulgareleaves are collected from a local garden, Baghdad, Iraq. The hot and cold aqueous extract was prepared to perform an In vitro antibacterial and antibiofilm actions .Antibacterial activities on various antibiotic resistant bacteria (Pseudomonas aeruginosa, Klebsiella pneumoniae, Echerichia coli, Staphylococcus aurous and staphylococcus epidermidis) were tested by well diffusion method. The biofilm-forming ability of the tested microorganisms was determined by the Microtiter-plate method. Fresh hot extract showed the highest effect and the dried cold extract had the lowest effect against gram-negative and gram-positive bacteria . fresh hot extract decrease significantly the biofilm formation and showed good ability as an antibiofilm to all strains that tested..Antimicrobial activity of the extract seemed to be related to the aqueous extract preparation method and that appeared by the difference between fresh and dried leave extract activity against pathogens and it can be used as potential antibiofilm agent against the biofilm forming strains.

Biosynthesized Silver Nanoparticles (AgNPs) from Trapa natans Peel Extract Exhibits Anti-Metastasis and Anti-Biofilm Potentials

Abstract: Background: This study deals with the anti-metastasis and anti-biofilm activities using silver nanoparticles (AgNPs) against human breast cancer cells (MCF-7) via in vitro methods. Different studies have been reported that biofilms formed by Klebsiella pneumoniae, Escherichia coli and Staphylococcus aureus are resistance to most of the recently used antibiotics due to MDR effect. Materials and methods: The characterization of biosynthesized particles was done by UV-vis spectroscopy, FTIR, SEM and XRD techniques. Cytotoxicity analysis was done by MTT assay and Neutral Red Uptake (NRU) assay. Anti-biofilm activity was carried out by using the bacterial strains of Klebsiella pneumoniae, Escherichia coli and Staphylococcus aureus respectively. Results: AgNPs exhibit strong absorption peak in the 420–500 nm range due to Surface Plasmon Resonance in UV- Vis spectroscopy. In FTIR analysis, we found phenol and alcohol compounds (at 3447 cm-1), carbonyl groups (at 1631 cm-1), alcoholic groups (at 1384 cm-1) and polysaccharides (at 1093 cm-1). In XRD that AgNPs formed were highly crystalline in nature. In SEM the diameter of AgNPs ranges from 30-90 nm and TEM depicts the size of AgNPs to be 15 ± 2 nm. T. natans-AgNPs exhibits strong cytotoxic activity against MCF-7 breast cancer cell line. Conclusion: The presence of different bioactive molecules in T.natans shows that, green synthesized AgNPs significantly inhibited proliferation of MCF-7 breast cancer cells by inducing apoptosis as well as antibiofilm activity using different bacterial colonies.For the very first time we are reporting the anticancer drugs repositioned as a potential antibiofilm agent in the Klebsiella pneumoniae, Escherichia coli and Staphylococcus aureus organisms. Key words: Trapa natans, Silver Nanoparticles (AgNPs),Human Breast Cancer Cells (MCF-7), Anti-biofilm, Anti-metastasis, Cytotoxicity. Key words: Trapa natans, Silver Nanoparticles (AgNPs),Human Breast Cancer Cells (MCF-7), Anti-biofilm, Anti-metastasis, Cytotoxicity.

Effect of Lactobacillus plantarum Biofilms on the Adhesion of Escherichia coli to Urinary Tract Devices.

Novel technologies to prevent biofilm formation on urinary tract devices (UTDs) are continually being developed, with the ultimate purpose of reducing the incidence of urinary infections. Probiotics have been described as having the ability to displace adhering uropathogens and inhibit microbial adhesion to UTD materials. This work aimed to evaluate the effect of pre-established Lactobacillus plantarum biofilms on the adhesion of Escherichia coli to medical-grade silicone. The optimal growth conditions of lactobacilli biofilms on silicone were first assessed in 12-well plates. Then, biofilms of L. plantarum were placed in contact with E. coli suspensions for up to 24 h under quasi-static conditions. Biofilm monitoring was performed by determining the number of culturable cells and by confocal laser scanning microscopy (CLSM). Results showed significant reductions of 76%, 77% and 99% in E. coli culturability after exposure to L. plantarum biofilms for 3, 6 and 12 h, respectively, corroborating the CLSM analysis. The interactions between microbial cell surfaces and the silicone surface with and without L. plantarum biofilms were also characterized using contact angle measurements, where E. coli was shown to be thermodynamically less prone to adhere to L. plantarum biofilms than to silicone. Thus, this study suggests the use of probiotic cells as potential antibiofilm agents for urinary tract applications.

Antioxidant and antibacterial study of 10 flavonoids revealed rutin as a potential antibiofilm agent in Klebsiella pneumoniae strains isolated from hospitalized patients

The emergence of multidrug resistance (MDR) and extensive drug resistance (XDR) in Klebsiella pneumoniae strains has posed great threats to conventional antibiotics. Previous studies have shown that plant-derived flavonoids have inhibitory functions against pathogens. However, in K. pneumoniae, the antibacterial activity of different flavonoids against growth and biofilm formation remains a mystery. The aim of the present study was to evaluate the antioxidant abilities of different flavonoids, to screen active ingredients and to identify their inhibitory effects on K. pneumoniae growth and biofilm formation. In total, 10 flavonoids representing 4 major categories were screened and used in this study. The antioxidant capacity of each flavonoid was evaluated through a DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Rutin showed the highest level of free radical scavenging capacity, followed by kaempferol, luteolin, quercetin, apigenin, hesperidin, sinensetin, naringenin, naringin and 3,5,6,7,8,3′,4′-heptamethoxyflavone. The inhibitory effects of rutin and naringin on bacterial growth were also compared. The lowest MICs of rutin were found against K. pneumoniae ATCC700603 (1024 μg/mL) and E. coli ATCC25922 (512 μg/mL). However, the MBICs were not found. Rutin showed strong inhibitory ability against both the growth curve and biofilm production. The expression profiles of 15 biofilm-related genes were analyzed in biofilm cells both with and without rutin treatment. The luxS gene and wabG gene were downregulated significantly by rutin treatment. Correlation analysis showed that mrkA gene expression was positively correlated with biofilm biomass accumulation. Our study indicated that biofilm production is correlated with the expression of several genes rather than one. MrkA gene expression was positively correlated with biofilm biomass accumulation. Our study screened rutin as a potential agent to inhibit K. pneumoniae biofilm formation.

Tryptophan interferes with the quorum sensing and cell surface hydrophobicity of Staphylococcus aureus: a promising approach to inhibit the biofilm development.

The online version contains supplementary material available at 10.1007/s13205-021-02924-3.

Identification, characterization and evaluation of novel antifungal cyclic peptides from Neobacillus drentensis

Marine microbes secrete exopolymeric substances (EPS), which surrounds the biofilm and inhibits the fungal growth. Elucidation of the structure and function of the extracellular exopolymeric substances is of vital relevance therapeutically. The active compound responsible for bioactivity was purified and characterized using TLC, LC/MS/MS, GC/MS and FT-IR. Bioactivity of the characterized cyclic peptides (CLPs) against azole resistant and susceptible Candida strains were examined for growth and biofilm formation using scanning electron microscopy, flow cytometry, confocal microscopy. In the present study we identified bioactive cyclic peptides from marine isolated Neobacillus drentensis that exhibited promising tensio-active properties and antifungal efficacy against azole resistant and susceptible Candida albicans. The cluster is composed of five CLP isoforms which were sequenced and identified as new peptides with compositional and structural variations in the amino acid sequence and fatty acid chain. In vitro cytotoxic activity of CLPs was tested in human fibroblast normal cells. We have observed that the CLPs repressed the Candida albicans growth and multiplication by inhibiting the biofilm formation and disruption of branching filamentous hyphae. CLPs have been found to arrest the C. albicans cell cycle by a block at G1-S transition followed by apoptotic cell death. The current studies suggest these natural marine derived CLPs function as potential anti-biofilm agents against azole C. albicans resistant strains.

Biosurfactant synergized with marine bacterial DNase disrupts polymicrobial biofilms.

Globally, the occurrence of biofilm associated infection has become an alarming menace to the medical fraternity because the thick exopolysaccharide layer encasing the biofilms makes the biofilm producing pathogens inherently resistant to antibiotics. Candida albicans, the most common pathogen among Candida spp. is the causative agent for superficial and invasive candidiasis. The morphological phase switching from yeast to hyphal form is one of the virulent traits of C. albicans critical for its pathogenicity. Owing to the emergence of antifungal resistance among this opportunistic fungus, there is a dire need for improvised alternative antifungal agents. In the present study, we have evaluated a biosurfactant from a marine bacterium for its biofilm disruption ability against C. albicans. This biosurfactant had the potential to disrupt biofilms as well as to inhibit the morphological transition from yeast to hyphae. In addition, this biosurfactant showed enhance disruption of mixed species biofilms of C. albicans and Staphylococcus epidermidis when combined with DNase isolated from marine bacteria. From the results obtained, it is evident that the biosurfactant could act as a potential antibiofilm agent against drug resistant C. albicans strains.

Antimicrobial, antibiofilm, antioxidant, anticancer, and phytochemical composition of the seed extract of Pongamia pinnata.

Ethyl acetate seed extract of Pongamia pinnata displayed the highest antimicrobial potential against all test pathogens and Staphylococcus epidermidis was reported as the most sensitive strain with MIC/MBC 1.56/3.12mgml-1. It inhibited S. epidermidis biofilm 97.43% at MIC and LM as well as FE-SEM micrographs displayed extensive disintegration in biofilm. It showed the highest TPC (1.23 ± 0.04g GAE g-1), TFC (0.95 ± 0.05g CE g-1), and antioxidant activity with IC50 18.47 ± 0.33μgml-1. MTT assay displayed concentration-dependent strong cytotoxicity on K562 cells on the treatment of ethyl acetate extract with an IC50 value of 84.41μgml-1. On the other hand, it showed minute cytotoxicity on normal PBMCs with an IC50 value of 410.14μgml-1. GC-MS analysis revealed that Hexadecanoic acid (35.97%); 2-(1,3-Benzodioxol-5-yl)furo[2,3-h]chromen-4-one (Pongaglabrone) (22.82%); 2,2-Dimethylindane-1,3-dione- (13.05%) were the three major components in ethyl acetate extract. The present investigation showcases ethyl acetate extract as a potent antimicrobial, antibiofilm, antioxidant, and anticancer agent that opens a new avenue for its phytochemicals as a therapeutic agent.

On-demand pH-sensitive surface charge-switchable polymeric micelles for targeting Pseudomonas aeruginosa biofilms development

Bacterial biofilm is the complicated clinical issues, which usually results in bacterial resistance and reduce the therapeutic efficacy of antibiotics. Although micelles have been drawn attention in treatment of the biofilms, the micelles effectively permeate and retain in biofilms still facing a big challenge. In this study, we fabricated on-demand pH-sensitive surface charge-switchable azithromycin (AZM)-encapsulated micelles (denoted as AZM-SCSMs), aiming to act as therapeutic agent for treating Pseudomonas aeruginosa (P. aeruginosa) biofilms. The AZM-SCSMs was composed of poly(l-lactide)-polyetherimide-hyd-methoxy polyethylene glycol (PLA-PEI-hyd-mPEG). It was noteworthy that the pH-sensitive acylhydrazone bond could be cleaved in acidic biofilm microenvironment, releasing the secondary AZM-loaded cationic micelles based on PLA-PEI (AZM-SCMs) without destroying the micellar integrity, which could tailor drug-bacterium interaction using micelles through electrostatic attraction. The results proved that positively charged AZM-SCMs could facilitate the enhanced penetration and retention inside biofilms, improved binding affinity with bacterial membrane, and added drug internalization, thus characterized as potential anti-biofilm agent. The excellent in vivo therapeutic performance of AZM-SCSMs was confirmed by the targeting delivery to the infected tissue and reduced bacterial burden in the abscess-bearing mice model. This study not only developed a novel method for construction non-depolymerized pH-sensitive SCSMs, but also provided an effective means for the treatment of biofilm-related infections.

In vitro anti-biofilm efficacy of sanguinarine against carbapenem-resistant Serratia marcescens

Sanguinarine, a plant-derived benzophenanthridine alkaloid, was studied in terms of its anti-biofilm effects against carbapenem-resistant Serratia marcescens (CRSM). Minimum inhibitory concentrations (MICs) and cell membrane integrity were measured to investigate the antimicrobial mechanism of sanguinarine. Additionally, the extent of biofilm formation by CRSM exposed to sanguinarine was measured by crystal violet staining and visualized via field emission scanning electron microscopy and confocal laser scanning microscopy. Sanguinarine displayed moderate activity against CRSM, with a MIC90 of 32 μg ml−1. Moreover, cell membrane integrity was severely disrupted by sanguinarine at 64 μg ml−1, and biofilm formation was sharply inhibited at 32 μg ml−1. The minimum biofilm eradication concentration was 512 μg ml−1 against mature CRSM biofilms. The overall results suggest that sanguinarine is a potential anti-biofilm agent that can be explored to treat CRSM infections.

Quercetin Inhibits Biofilm Formation by Decreasing the Production of EPS and Altering the Composition of EPS in Staphylococcus epidermidis

Staphylococcus epidermidis is an opportunistic pathogen, and its biofilm formation ability is an important virulent factor. Quercetin, a typical flavonoid ubiquitously used in dietary supplementation, is known for its antioxidant property, but its anti-biofilm activity against S. epidermidis remains unknown. In this study, the anti-biofilm activity of quercetin was investigated using S. epidermidis ATCC35984, a strong biofilm-positive strain. An attempt was made to disclose the mechanisms of the anti-biofilm activity of quercetin. S. epidermidis exhibited a less cell surface hydrophobicity after quercetin treatment. Also, quercetin effectively inhibited S. epidermidis cells from adhering to the glass slides. Quercetin downregulated the intercellular adhesion (ica) locus and then polysaccharide intercellular adhesin (PIA) production was reduced. Therefore, S. epidermidis cells became less hydrophobic, which supported quercetin’s anti-biofilm effect. Our study suggests that quercetin from plants be given further attention as a potential anti-biofilm agent against the biofilm formation of S. epidermidis, even biofilm infections of other bacteria.

Biofabricated Fatty Acids-Capped Silver Nanoparticles as Potential Antibacterial, Antifungal, Antibiofilm and Anticancer Agents.

The current study demonstrates the synthesis of fatty acids (FAs) capped silver nanoparticles (AgNPs) using aqueous poly-herbal drug Liv52 extract (PLE) as a reducing, dispersing and stabilizing agent. The NPs were characterized by various techniques and used to investigate their potent antibacterial, antibiofilm, antifungal and anticancer activities. GC-MS analysis of PLE shows a total of 37 peaks for a variety of bio-actives compounds. Amongst them, n-hexadecanoic acid (21.95%), linoleic acid (20.45%), oleic acid (18.01%) and stearic acid (13.99%) were found predominately and most likely acted as reducing, stabilizing and encapsulation FAs in LIV-AgNPs formation. FTIR analysis of LIV-AgNPs shows some other functional bio-actives like proteins, sugars and alkenes in the soft PLE corona. The zone of inhibition was 10.0 ± 2.2–18.5 ± 1.0 mm, 10.5 ± 2.5–22.5 ± 1.5 mm and 13.7 ± 1.0–16.5 ± 1.2 against P. aeruginosa, S. aureus and C. albicans, respectively. LIV-AgNPs inhibit biofilm formation in a dose-dependent manner i.e., 54.4 ± 3.1%—10.12 ± 2.3% (S. aureus), 72.7 ± 2.2%–23.3 ± 5.2% (P. aeruginosa) and 85.4 ± 3.3%–25.6 ± 2.2% (C. albicans), and SEM analysis of treated planktonic cells and their biofilm biomass validated the fitness of LIV-AgNPs in future nanoantibiotics. In addition, as prepared FAs rich PLE capped AgNPs have also exhibited significant (p < 0.05 *) antiproliferative activity against cultured HCT-116 cells. Overall, this is a very first demonstration on employment of FAs rich PLE for the synthesis of highly dispersible, stable and uniform sized AgNPs and their antibacterial, antifungal, antibiofilm and anticancer efficacy.

Antibiotic Efficacy Testing in an Ex vivo Model of Pseudomonas aeruginosa and Staphylococcus aureus Biofilms in the Cystic Fibrosis Lung.

The effective prescription of antibiotics for the bacterial biofilms present within the lungs of individuals with cystic fibrosis (CF) is limited by a poor correlation between antibiotic susceptibility testing (AST) results using standard diagnostic methods (e.g., broth microdilution, disk diffusion, or Etest) and clinical outcomes after antibiotic treatment. Attempts to improve AST by the use of off-the-shelf biofilm growth platforms show little improvement in results. The limited ability of in vitro biofilm systems to mimic the physicochemical environment of the CF lung and, therefore bacterial physiology and biofilm architecture, also acts as a brake on the discovery of novel therapies for CF infection. Here, we present a protocol to perform AST of CF pathogens grown as mature, in vivo-like biofilms in an ex vivo CF lung model comprised of pig bronchiolar tissue and synthetic CF sputum (ex vivopig lung, EVPL). Several in vitroassays exist for biofilm susceptibility testing, using either standard laboratory medium or various formulations of synthetic CF sputum in microtiter plates. Both growth medium and biofilm substrate (polystyrene plate vs. bronchiolar tissue) are likely to affect biofilm antibiotic tolerance. We show enhanced tolerance of clinical Pseudomonas aeruginosa and Staphylococcus aureus isolates in the ex vivo model; the effects of antibiotic treatment of biofilms is not correlated with the minimum inhibitory concentration (MIC) in standard microdilution assays or a sensitive/resistant classification in disk diffusion assays. The ex vivo platform could be used for bespoke biofilm AST of patient samples and as an enhanced testing platform for potential antibiofilm agents during pharmaceutical research and development. Improving the prescription or acceleration of antibiofilm drug discovery through the use of more in vivo-like testing platforms could drastically improve health outcomes for individuals with CF, as well as reduce the costs of clinical treatment and discovery research.

Antibiotic Efficacy Testing in an Ex vivo Model of &lt;em&gt;Pseudomonas aeruginosa&lt;/em&gt; and &lt;em&gt;Staphylococcus aureus&lt;/em&gt; Biofilms in the Cystic Fibrosis Lung

The effective prescription of antibiotics for the bacterial biofilms present within the lungs of individuals with cystic fibrosis (CF) is limited by a poor correlation between antibiotic susceptibility testing (AST) results using standard diagnostic methods (e.g., broth microdilution, disk diffusion, or Etest) and clinical outcomes after antibiotic treatment. Attempts to improve AST by the use of off-the-shelf biofilm growth platforms show little improvement in results. The limited ability of in vitro biofilm systems to mimic the physicochemical environment of the CF lung and, therefore bacterial physiology and biofilm architecture, also acts as a brake on the discovery of novel therapies for CF infection. Here, we present a protocol to perform AST of CF pathogens grown as mature, in vivo-like biofilms in an ex vivo CF lung model comprised of pig bronchiolar tissue and synthetic CF sputum (ex vivo pig lung, EVPL). Several in vitro assays exist for biofilm susceptibility testing, using either standard laboratory medium or various formulations of synthetic CF sputum in microtiter plates. Both growth medium and biofilm substrate (polystyrene plate vs. bronchiolar tissue) are likely to affect biofilm antibiotic tolerance. We show enhanced tolerance of clinical Pseudomonas aeruginosa and Staphylococcus aureus isolates in the ex vivo model; the effects of antibiotic treatment of biofilms is not correlated with the minimum inhibitory concentration (MIC) in standard microdilution assays or a sensitive/resistant classification in disk diffusion assays. The ex vivo platform could be used for bespoke biofilm AST of patient samples and as an enhanced testing platform for potential antibiofilm agents during pharmaceutical research and development. Improving the prescription or acceleration of antibiofilm drug discovery through the use of more in vivo-like testing platforms could drastically improve health outcomes for individuals with CF, as well as reduce the costs of clinical treatment and discovery research.

Optimization of biosurfactant production by marine Streptomyces youssoufiensis SNSAA03: A comparative study of RSM and ANN approach

The aim of the present study was to optimize media components to increase the production of biosurfactant (Bs) by Streptomyces youssoufiensis SNSAA03 employing response surface methodology (RSM) and artificial neural network (ANN). The RSM showed the following as the optimal condition for the production of Bs: glucose 15 g L–1, casein 0.3 g L–1, NaCl 2.25 g L–1 and pH 7. The superiority of the ANN model was proved with more accurate and consistent R2 values 0.998 when compared to RSM R2 value 0.980. The Bs produced by SNSAA03 was identified as glycolipid with 4–methoxy–2, 5–dimethylfuran–3(2H)–one and 1–Nanodecene acid. The micro titre plate assay showed that glycolipid at the concentration of 125 µg mL−1 was effective in controlling the biofilm formation by S. aureus. Thus, the glycolipid produced from SNSAA03 can be used as a potent antibiofilm agent.

Quorum Sensing Inhibitory Activities of Oridonin in Pseudomonas Aeruginosa

Pseudomonas aeruginosa is an opportunistic pathogen, and it can produce cell-associated and extracellular virulence factors. Several of these virulence factors have been demonstrated to be regulated by quorum sensing (QS). Disabling QS system with anti-infective agent is considered as a potential strategy to prevent bacterial infection. Rabdosia rubescens has been used as antibacterial agents for many centuries in China. In this study, Oridonin, the major active components of Rabdosia rubescens, was tested for QS inhibition in Pseudomonas aeruginosa. QS inhibitory activity is demonstrated by reduction in pyocyanin (58.4%), rhamnolipids (64.3%), elastase (58.6%), and protease (49.1%) in Pseudomonas aeruginosa PAO1 at 125 µg/ml (MIC) concentration. Biofilm formation by Pseudomonas aeruginosa PAO1 was reduced considerably (40.3-57.7%) over control. These findings suggest that Oridonin might be a potent Quorum Sensing Inhibitor (QSI) and anti-biofilm agent in the treatment of Pseudomonas aeruginosa infections.

AHL-Based QS Modulation and Inhibition of Biofilm Forming Foot Ulcer Pathogens by Selected Medicinal Plants

The crisis of antibiotic resistance necessitates the search of phytochemicals as potential antimicrobials, anti-quorum sensing and antibiofilm agents. For the present study, fifteen (15) medicinal plants were evaluated for biological activities against multi-drug resistant (MDR) pathogenic bacteria (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis), associated with diabetic foot ulcer. Antibacterial activities revealed noteworthy minimum inhibitory concentration (MIC) values ≤ 1 mg/mL for thirteen (13) out of the sixty (60) plant extracts screened. The potent extracts included Euclea natalensis ethyl acetate (0.25 mg/mL), Aloe ferox methanol (0.5 mg/ml) and Warburgia salutaris aqueous (0.5 mg/mL) extracts. Chemical profiling of the active extracts using gas chromatography-mass spectrometry (GC-MS) identified neophytadiene, guanosine, squalene, cis megastigma-5,8-diene-4-one and sorbitol as prevalent compounds. Anti-quorum sensing activities of E. natalensis (ethyl acetate), A. ferox (methanol) and W. salutaris (aqueous) extracts ranged from 4.81 - 58.34% with E. natalensis (ethyl-acetate) showing the highest activity. Molecular docking against CviR protein showed selected compounds having high docking scores, with sorbitol showing the highest score of -7.04 kcal/mol. Warburgia salutaris aqueous extract exhibited the highest biofilm inhibition (73%) against E. coli. Euclea natalensis, Aloe ferox and Warburgia salutaris compounds act as antagonists of AHL signaling, thus may serve as candidates in antipathogenic and antibiofilm phytomedicine development for drug-resistant foot ulcer pathogens.

Octominin: An antibacterial and anti-biofilm peptide for controlling the multidrug resistance and pathogenic Streptococcus parauberis

Streptococcus parauberis is a pathogenic gram-positive bacterium that causes streptococcosis infection in fish. Since S. parauberis is becoming resistant to multiple antibiotics, the development of alternatives, such as antimicrobial peptides, has gained great attention. Octominin, derived from the defense protein of Octopus minor, showed a significant antimicrobial activity against multidrug resistance S. parauberis, with a minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of 50 and 100 μg/mL, respectively. Furthermore, time-kill kinetics, agar diffusion, and bacterial viability assays confirmed the concentration-dependent antibacterial activity of Octominin against S. parauberis. Field emission scanning electron microscopy analysis showed morphological and ultra-structural changes in S. parauberis upon Octominin treatment. Moreover, Octominin treatment demonstrated changes in membrane permeability, induced reactive oxygen species (ROS), and its binding ability to genomic DNA, suggesting its strong bactericidal activity with multiple modes of action. We confirmed the inhibition of biofilm formation and the eradication of existing biofilms in a concentration-dependent manner. Additionally, Octominin on S. parauberis at transcriptional level exhibited downregulation of membrane formation (pgsA and cds1), DNA repairing (recF), biofilm formation (pgaB and epsF) genes, while upregulation of ROS detoxification (sodA) and DNA protecting (ahpF) related genes. An in vivo study confirmed a significantly (P < 0.05) higher relative percentage survival in Octominin-treated larval zebrafish exposed to S. parauberis (93.3%) compared to the control group (20.0%). Collectively, our results confirm that Octominin could be a potential antibacterial and anti-biofilm agent against S. parauberis.

Inhibition of biofilm and quorum sensing-regulated virulence factors in Pseudomonas aeruginosa by Cuphea carthagenensis (Jacq.) J. F. Macbr. Leaf extract: An in vitro study

Ethnopharmcological relevanceMicrobial biofilm formation, a quorum sensing (QS) regulated process, is one of the major causes of nosocomial and chronic infections, foodborne diseases, and associated deaths. Various approaches have been used to eradicate the menace of biofilm. Ethnomedicinal plants as potent antibiofilm agents are gaining a lot of interest in an era where the drug resistance is increasing and the availability of potent antibiotics is no longer promised. In this context, the methanol extract of Cuphea carthagenensis (CCMD), an ethno-medicinal and culinary herb, was evaluated as an antibiofilm and anti-QS agent against Pseudomonas aeruginosa.Aim of the study: The aim of the study is to evaluate the antibiofilm and anti-QS activity of an ethnomedicinal plant against a strong biofilm forming microorganism, P. aeruginosa. MethodsAntibiofilm activity of CCMD was demonstrated at different concentrations by Tissue Culture Plate, Test Tube method and other microscopic techniques. The effect of CCMD on QS and QS-related virulence factors viz. Pyocyanin, exopolymeric substance matrix (EPS), total protease, elastase, pyoverdin and swimming motility in P. aeruginosa were also evaluated. Antioxidant activity (DPPH & FRAP), total phenolic and flavonoid content were also checked. In order to determine the composition of the extract HPLC analysis was also performed. ResultsIn vitro study demonstrated a significant inhibition of biofilm formation (81.88 ± 2.57%) as well as production of QS-dependent virulence factors in P. aeruginosa. The extract also inhibited violacein production (83.31 ± 2.77%) in Chromobacterium violaceum which correlates with the reduction in QS-mediated virulence factors. The extract showed 64.79% ± 0.83% DPPH scavenging activity and reduction of ferricyanide complex (Fe3+) to the ferrous form (Fe2+) in DPPH and FRAP assay, respectively. Furthermore, the extract showed thermal stability and does not have any growth inhibitory effect on P. aeruginosa. The HPLC analysis demonstrated the presence of ellagic acid, ascorbic acid and hippuric acid in the extract. ConclusionThis work is the first to demonstrate that C. carthagenensis can attenuate biofilm formation and QS-mediated virulence factors of P. aeruginosa. Further investigation is required to use this ethnomedicinal plant (CCMD) as an important source of antibiofilm agents.

Laser-Irradiated Chlorpromazine as a Potent Anti-Biofilm Agent for Coating of Biomedical Devices

Nowadays, antibiotic resistance has become increasingly common, triggering a global health crisis, immediately needing alternative, including repurposed drugs with potent bactericidal effects. We demonstrated that chlorpromazine aqueous solutions exposed to laser radiation exhibited visible activity against various microorganisms. The aim of this study was to investigate the quantitative antimicrobial activity of chlorpromazine in non-irradiated and 4-h laser irradiated form. Also, we examined the effect of both solutions impregnated on a cotton patch, cannula, and urinary catheter against Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa and Escherichia coli. In all experimental versions, the chlorpromazine antimicrobial activity was enhanced by laser exposure. Besides the experimental results, the in silico analyses using molecular docking proved that the improved antimicrobial activity of the irradiated compound was a result of the combined action of the photoproducts on the biological target (s). Our results show that laser radiation could alter the molecular structure of various drugs and their effects, proving to be a promising strategy to halt antibiotic resistance, by repurposing current medicines for new antimicrobial strategies, thereby decreasing the costs and time for the development of more efficient drugs.