Detectable quorum signaling molecule via PANI-metal oxides nanocomposites sensors.

The detection of N-hexanoyl-l-homoserine lactone (C6-HSL), a crucial signal in Gram-negative bacterial communication, is essential for addressing microbiologically influenced corrosion (MIC) induced by sulfate-reducing bacteria (SRB) in oil and gas industries. Metal oxides (MOx) intercalated into conducting polymers (CPs) offer a promising sensing approach due to their effective detection of biological molecules such as C6-HSL. In this study, we synthesized and characterized two MOx/polyaniline-dodecyl benzene sulfonic acid (PANI-DBSA) nanocomposites, namely ZnO/PANI-DBSA and Fe2O3/PANI-DBSA. These nanocomposites were applied with 1% by-weight carbon paste over a carbon working electrode (WE) for qualitative and quantitative detection of C6-HSL through electrochemical analysis. The electrochemical impedance spectroscopy (EIS) confirmed the composites' capability to monitor C6-HSL produced by SRB-biofilm, with detection limits of 624 ppm for ZnO/PANI-DBSA and 441 ppm for Fe2O3/PANI-DBSA. Furthermore, calorimetric measurements validated the presence of SRB-biofilm, supporting the EIS analysis. The utilization of these MOx/CP nanocomposites offers a practical approach for detecting C6-HSL and monitoring SRB-biofilm formation, aiding in MIC management in oil and gas wells. The ZnO/PANI-DBSA-based sensor exhibited higher sensitivity towards C6-HSL compared to Fe2O3/PANI-DBSA, indicating its potential for enhanced detection capabilities in this context. Stability tests revealed ZnO/PANI-DBSA's superior stability over Fe2O3/PANI-DBSA, with both sensors retaining approximately 85-90% of their initial current after 1 month, demonstrating remarkable reproducibility and durability.

Enhancing physical characteristics and antibacterial efficacy of chitosan through investigation of microwave-assisted chemically formulated chitosan-coated ZnO and chitosan/ZnO physical composite.

This study investigates the creation and analysis of chitosan-zinc oxide (CS-ZnO) nanocomposites, exploring their effectiveness in inhibiting bacteria. Two synthesis approaches, physical and chemical, were utilized. The CS-ZnO nanocomposites demonstrated strong antibacterial properties, especially against Staphylococcus aureus, a Gram-positive bacterium. Chemically synthesized nanocomposites (CZ10 and CZ100) exhibited larger inhibition zones (16.4 mm and 18.7 mm) compared to physically prepared CS-Z5 and CS-Z20 (12.2 mm and 13.8 mm) against Staphylococcus aureus. Moreover, CZ nanocomposites displayed enhanced thermal stability, with decomposition temperatures of 281°C and 290°C, surpassing CS-Z5 and CS-Z20 (260°C and 258°C). The residual mass percentages at 800°C were significantly higher for CZ10 and CZ100 (58% and 61%) than for CS-Z5 and CS-Z20 (36% and 34%). UV-Visible spectroscopy revealed reduced band gaps in the CS-ZnO nanocomposites, indicating improved light absorption. Transmission electron microscopy (TEM) confirmed uniform dispersion of ZnO nanoparticles within the chitosan matrix. In conclusion, this research underscores the impressive antimicrobial potential of CS-ZnO nanocomposites, especially against Gram-positive bacteria, and highlights their enhanced thermal stability. These findings hold promise for diverse applications in industries such as medicine, pharmaceuticals, and materials science, contributing to the development of sustainable materials with robust antimicrobial properties.

Chitosan-Aspirin Combination Inhibits Quorum-Sensing Synthases (lasI and rhlI) in Pseudomonas aeruginosa.

BACKGROUND: Quorum sensing (QS) controls the virulence of P. aeruginosa. This study aims to determine the anti-QS activity of aspirin alone and in combination with chitosan to reach maximum inhibition. We tested ten virulent Pseudomonas aeruginosa (P. aeruginosa) isolates and screened for N-acyl homoserine lactone (AHL) production using Agrobacterium tumefaciens as a biosensor. P. aeruginosa isolates were treated with sub-minimum inhibitory concentrations (MICs) of aspirin and chitosan-aspirin. We used broth microdilution and checkerboard titration methods to determine the MICs and the synergistic effect of these two compounds, respectively. Real-time polymerase chain reaction (PCR) was used to estimate the anti-QS activity of the aspirin-chitosan combination on the expression of lasI and rhlI genes. RESULTS: Aspirin decreased the motility and production of AHLs, pyocyanin, and biofilm. Chitosan potentiated the inhibitory effect of aspirin. The chitosan-aspirin combination inhibited lasI and rhlI gene expression in PAO1 (ATCC 15692) by 7.12- and 0.92-fold, respectively. In clinical isolates, the expression of lasI and rhlI was decreased by 1.76 × 102- and 1.63 × 104-fold, respectively. Molecular docking analysis revealed that aspirin could fit into the active sites of the QS synthases lasI and rhlI with a high binding affinity, causing conformational changes that resulted in their inhibition. CONCLUSIONS: The chitosan-aspirin combination provides new insights into treating virulent and resistant P. aeruginosa.

Endophytic Aspergillus hiratsukae mediated biosynthesis of silver nanoparticles and their antimicrobial and photocatalytic activities.

In the current study, endophytic Aspergillus hiratsukae was used for the biosynthesis of silver nanoparticles (Ag-NPs) for the first time. The characterizations were performed using X ray diffraction (XRD), Transmission electron microscopy (TEM), Scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX), Dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), and UV-Vis spectroscopy. The obtained results demonstrated the successful formation of crystalline, spherical Ag-NPs with particle diameters ranging from 16 to 31 nm. The FT-IR studied and displayed the various functional groups involved, which played a role in capping and reducing agents for Ag-NPs production. The SEM-EDX revealed that the main constituent of the AS-formed sample was primarily Ag, with a weight percentage of 64.2%. The mycosynthesized Ag-NPs were assessed for antimicrobial as well as photocatalytic activities. The antimicrobial results indicated that the synthesized Ag-NPs possess notable antibacterial efficacy against Staphylococcus aureus, Bacillus subtilis, and Escherichia coli, with minimum inhibitory concentrations (MICs) of Ag-NPs ranging from 62.5 to 250 µg/mL. Moreover, the biosynthesized Ag-NPs demonstrated weak antifungal activity against Aspergillus brasiliensis and Candida albicans, with MICs of 500 and 1,000 µg/mL, respectively. In addition, the mycosynthesized Ag-NPs exhibited photocatalytic activity toward acid black 2 (nigrosine) dye under both light and dark stimulation. Notably, After 300 min exposure to light, the nigrosine dye was degraded by 93%. In contrast, 51% degradation was observed after 300 min in darkness. In conclusion, Ag-NPs were successfully biosynthesized using endophytic A. hiratsukae and also exhibited antimicrobial and photocatalytic activities that can be used in environmental applications.

Modulating the transcriptomic profile of multidrug-resistant Klebsiella pneumoniae biofilm formation by antibiotics in combination with zinc sulfate.

BACKGROUND: Klebsiella pneumoniae is a significant healthcare-associated pathogen. We investigated the antimicrobial interaction pattern between zinc sulfate and antibiotics against K. pneumoniae biofilm on the phenotypic and genotypic levels. METHODS: Determining the minimum biofilm inhibitory concentrations and the transcriptomic profile of K. pneumoniae biofilm formation genes post-treatment were carried out to evaluate the effect on the phenotypic and genotypic levels, respectively. RESULTS: Zinc enhanced the antibiofilm potentials of cephalosporins, aminoglycosides, and ertapenem, whereas it antagonizes the effectiveness of fluoroquinolones and meropenem on the phenotypic level. On the molecular level, zinc enhanced the anti-biofilm efficacies of cephalosporins (cefotaxime, ceftriaxone, ceftazidime, cefpirome, and cefepime) via down-regulating the expression of biofilm-related genes by 18-, 38-, 5-, 77- and 2-folds, respectively. Zinc in combination with aminoglycosides (kanamycin, gentamicin, and amikacin) reduced the expression of biofilm-related genes by 40-, 2602- and 20-folds, respectively, and by 2-folds in combination with ertapenem. However, a reduction in the down-regulatory potentials of fluoroquinolones was recorded following combination with zinc by 2-, 2-, 15- and 14-folds, respectively, and an up-regulation in the expression levels of the tested genes by 2-folds in the case of zinc/meropenem combination. CONCLUSIONS: Results revealed variable interaction patterns between different antibiotics in combination with zinc. Current findings also shed light on the antibiofilm potentials of zinc/antibiotics combinations especially when combining zinc with fluoroquinolones or meropenem to avoid their antagonistic effects.

Evaluation of the Cariogenic and Anti-Cariogenic Potential of Human Colostrum and Colostrum-Derived Probiotics: Impact on S. mutans Growth, Biofilm Formation, and L. rhamnosus Growth.

Human colostrum (HC) is essential for oral health as it is rich in probiotics that could affect the growth of the cariogenic S. mutans and its biofilm formation; hindering dental caries in advance. In this study, HC was collected from 36 healthy mothers 1-3 days postpartum. The effect of HC on oral health was carried out by assessing the impact of HC and its derived probiotics' cell-free supernatants (CFS) on the growth of S. mutans (using modified well diffusion) and its biofilm formation (using microtiter plate assay). Moreover, the effect of whole HC on L. rhamnosus, a probiotic oral bacterium, was examined. Probiotics were isolated and identified phenotypically by API 50 CH carbohydrate fermentation and genotypically by 16S rRNA amplification. The in vitro study revealed that HC has cariogenic activity and is associated with biofilm formation. Biofilm strength was inversely proportional to HC dilution (p-value < 0.0001). Nevertheless, HC and colostrum-derived probiotics improve oral health by inhibiting the growth of caries-inducing S. mutans with lower inhibition to L. rhamnosus probiotics. The CFS of isolated probiotics reduced the biofilm formation via the cariogenic S. mutans. These results are not only promising for caries eradication, but they also highlight the importance of breastfeeding infants from their first hours to shape healthy oral microbiota, protecting them from various diseases including dental caries.

Mathematical pharmacodynamic modeling for antimicrobial assessment of ceftazidime/colistin versus gentamicin/meropenem combinations against carbapenem-resistant Pseudomonas aeruginosa biofilm.

BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) represents an escalating healthcare hazard with high mortality worldwide, especially in presence of biofilm. The current study aimed to evaluate the anti-biofilm potentials of ceftazidime, colistin, gentamicin, and meropenem alone and in combinations against biofilm-forming CRPA. METHODS: Biofilm killing and checkerboard assay were performed to detect the effectiveness of combined antibiotics against biofilms and planktonic cells, respectively. The bacterial bioburden retrieved from the established biofilms following treatment with combined antibiotics was utilized to construct a three-dimensional response surface plot. A sigmoidal maximum effect model was applied to determine the pharmacodynamic parameters (maximal effect, median effective concentration, and Hill factor) of each antibiotic to create a mathematical three-dimensional response surface plot. RESULTS: Data revealed statistically significant (p < 0.05) superior anti-biofilm potential in the case of colistin followed by a lower effect in the case of gentamicin and meropenem, while ceftazidime exhibited the least anti-biofilm activity. The fractional inhibitory concentration index (FICI ≤ 0.5) indicated synergism following treatment with the combined antibiotics. An elevated anti-biofilm activity was recorded in the case of gentamicin/meropenem compared to ceftazidime/colistin. Synergistic anti-biofilm potentials were also detected via the simulated pharmacodynamic modeling, with higher anti-biofilm activity in the case of the in vitro observation compared to the simulated anti-biofilm profile. CONCLUSIONS: The present study highlighted the synergistic potentials of the tested antibiotic combinations against P. aeruginosa biofilms and the importance of the mathematical pharmacodynamic modeling in investigating the efficacy of antibiotics in combination as an effective strategy for successful antibiotic therapy to tackle the extensively growing resistance to the currently available antibiotics.

Molecular docking and nucleotide sequencing of successive expressed recombinant fungal peroxidase gene in E.coli.

BACKGROUND: Fungal peroxidases are oxidoreductases that utilize hydrogen peroxide to catalyze lignin biodegradation. RESULTS: PER-K (peroxidase synthesis codon gene) was transformed from Aspergillus niger strain AN512 deposited in the National Center for Biotechnology Information with the accession number OK323140 to Escherichia coli strain (BL21-T7 with YEp356R recombinant plasmid) via calcium chloride heat-shock method. The impact of four parameters (CaCl2 concentrations, centrifugation time, shaking speed, growth intensity) on the efficacy of the transformation process was evaluated. Furthermore, peroxidase production after optimization was assessed both qualitatively and quantitatively, as well as SDS-PAGE analysis. The optimum conditions for a successful transformation process were as follows: CaCl2 concentrations (50 mM), centrifugation time (20 min), shaking speed (200 rpm), and growth optical density (0.45). PCR and gel electrophoresis detect DNA bands with lengths 175, 179, and 211 bps corresponding to UA3, AmpR, and PER-K genes respectively besides partially sequencing the PER-K gene. Pyrogallol/hydrogen peroxide assay confirmed peroxidase production, and the activity of the enzyme was determined to be 3924 U/L. SDS-PAGE analysis also confirms peroxidase production illustrated by the appearance of a single peroxidase protein band after staining with Coomassie blue R-250. CONCLUSION: A successful peroxidase-gene (PER-K) transformation from fungi to bacteria was performed correctly. The enzyme activity was screened, and partial sequencing of PER-K gene was analyzed successively. The protein 3D structure was generated via in silico homology modeling, and determination of binding sites and biological annotations of the constructed protein were carried out via COACH and COFACTOR based on the I-TASSER structure prediction.

Green Biosynthesis of Selenium Nanoparticles Using Orange Peel Waste: Characterization, Antibacterial and Antibiofilm Activities against Multidrug-Resistant Bacteria.

There is an increase of pathogenic multidrug-resistant bacteria globally due to the misuse of antibiotics. Recently, more scientists used metal nanoparticles to counteract antibacterial resistance. In this study, orange peel waste (OPW) was used for selenium nanoparticles' (Se-NPs) biosynthesis through the green and ecofriendly method, and their applications as antibacterial and antibiofilm agents. Green biosynthesized Se-NPs were characterized using FTIR, XRD, SEM, EDAX, and TEM. Characterization results revealed that biosynthesized Se-NPs were highly crystalline, spherical, and polydisperse, and had sizes in the range of 16-95 nm. The biosynthesized Se-NPs were evaluated as antibacterial and antibiofilm activities against multidrug-resistant bacteria. Results illustrated that Se-NPs exhibited potential antibacterial activity against Gram-positive bacteria (S. aureus ATCC 29213 and biofilm-producing clinical isolates of S. aureus) and Gram-negative bacteria (Pseudomonas aeruginosa PAO1, MDR, biofilm, and quorum-sensing and producing clinical isolates of MDR P. aeruginosa, MDR E. coli, and K. pneumonia). Moreover, results illustrated that S. aureus ATCC 29213 was the most sensitive bacteria to Se-NPs at 1000 µg/mL, where the inhibition zone was 35 mm and MIC was 25 µg/mL. Furthermore, Se-NPs at 0.25 and 0.5 MIC decreased the biofilm significantly. The largest inhibition of biofilm was noticed in MDR K. pneumonia, which was 62% and 92% at 0.25 and 0.5 MIC, respectively. In conclusion, Se-NPs were successfully biosynthesized using OPW through the green method and had promising antibacterial and antibiofilm activity against multidrug-resistant bacteria, which can be used later in fighting resistant bacteria.

Chitosan Silver and Gold Nanoparticle Formation Using Endophytic Fungi as Powerful Antimicrobial and Anti-Biofilm Potentialities.

Nanotechnology is emerging as a new technology with encouraging innovations. Global antibiotic use has grown enormously, with antibiotic resistance increasing by about 80 percent. In view of this alarming situation, intensive research has been carried out into biogenic nanoparticles and their antibacterial, antifungal, and antitumor activities. Many methods are available to enhance stability and dispersion via peroration of conjugate with a polymer, such as chitosan, and other bioactive natural products. Two marine fungi were isolated and identified as Aspergillus sp. and Alternaria sp. via sequencing of the 16S rRNA gene. In this work, these strains were used to form the conjugation of biogenic silver nanoparticles (AgNPs) from Aspergillus sp. Silv2 extract and gold nanoparticles (AuNPs) from Alternaria sp. Gol2 extracts with chitosan to prepare chitosan-AgNPs and chitosan-AuNP conjugates. A variety of imaging and analytical methods, such as UV-vis, X-ray powder diffraction (XRD), FTIR spectroscopy, transmission electron microscopy (TEM), and scanning electron microscopy (SEM) were utilized to characterize biogenic nanoparticles and conjugates. The biosynthesized Ag and Au nanoparticles along with the prepared conjugates were evaluated for their antimicrobial effects on Gram-negative and Gram-positive bacterial isolates, including Escherichia coli and Staphylococcus aureus. Both chitosan-AgNP and AuNP showed powerful antimicrobial activities compared to the control. On the other hand, chitosan-AgNP conjugation had better antibacterial ctivity than chitosan-AuNPs, which exhibited moderate activity against S. aureus and very low activity against E. coli. Furthermore, the antibiofilm potentials of the prepared conjugates were tested against four biofilm-forming bacteria, including P. aeruginosa, B. subtilis, E. coli, and S. aureus. The obtained results indicate that the chitosan-AgNP showed a promising anti-biofilm activities on all strains, especially S. aureus, while chitosan-AuNP conjugates showed moderate anti-biofilm against B. subtilis and weak activities against the other three strains. These results showed the superiority of chitosan-AgNP as a promising antibacterial as well as biofilm formation inhibitors.

The Chemical Profiling, Docking Study, and Antimicrobial and Antibiofilm Activities of the Endophytic fungi Aspergillus sp. AP5.

Growing data suggest that Aspergillus niger, an endophytic fungus, is a rich source of natural compounds with a wide range of biological properties. This study aimed to examine the antimicrobial and antibiofilm capabilities of the Phragmites australis-derived endophyte against a set of pathogenic bacteria and fungi. The endophytic fungus Aspergillus sp. AP5 was isolated from the leaves of P. australis. The chemical profile of the fungal crude extract was identified by spectroscopic analysis using LC-HRESIMS. The fungal-derived extract was evaluated for its antimicrobial activity towards a set of pathogenic bacterial and fungal strains including Staphylococcus aureus, Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella sp., Candida albicans, and Aspergillus niger. Moreover, antibiofilm activity toward four resistant biofilm-forming bacteria was also evaluated. Additionally, a neural-networking pharmacophore-based visual screening predicted the most probable bioactive compounds in the obtained extract. The AP5-EtOAc extract was found to have potent antibacterial activities against S. aureus, E. coli, and Klebsiella sp., while it exhibited low antibacterial activity toward P. Vulgaris and P. aeruginosa and displayed anticandidal activity. The AP5-EtOAc extract had significant antibiofilm activity in S. aureus, followed by P. aeruginosa. The active metabolites' antifungal and/or antibacterial activities may be due to targeting the fungal CYP 51 and/or the bacterial Gyr-B.

Biofilm Inhibitory Activity of Actinomycete-Synthesized AgNPs with Low Cytotoxic Effect: Experimental and In Silico Study.

The emergence of resistance by biofilm-forming bacteria has reached alarming and dangerous levels that threaten human civilization. The current study sought to investigate the antibiofilm potential of green-synthesized silver nanoparticles, mediated by a new Streptomyces strain. Zeta potential, transmission electron microscopy (TEM), and UV-Vis spectroscopy were used to analyze the biosynthesized AgNPs. Results revealed that silver nanoparticles had a size of (5.55 and 45.00 nm) nm and a spherical shape, with surface plasmon resonance (SPR) absorption at 400-460 nm in the UV-vis spectra establishing the formation of Streptomyces-Ag-NPs. The biosynthesized AgNPs showed a pronounced antibacterial efficacy against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. Moreover, the obtained Streptomyces-AgNPs exerted biofilm inhibition activity against nosocomial hospital-resistant bacteria, including Bacillus subtilis, Staphylococcus aureus, and Escherichia coli. The mechanism of biogenic AgNPs antibacterial action was visualized using TEM, which indicated the AgNPs accumulation and disruption of bacterial cell membrane function. Additionally, a molecular docking study was conducted to evaluate the binding mode of AgNPs with an Escherichia coli outer membrane. Furthermore, the cytotoxic profile of the AgNPs was evaluated toward three cell lines (MCF-7, HepG2 & HCT 116), and the low cytotoxic effects of the obtained nanoparticles indicated their possible medical application with low risks to human health.

Chitosan and chitosan-zinc oxide nanocomposite inhibit expression of LasI and RhlI genes and quorum sensing dependent virulence factors of Pseudomonas aeruginosa.

The great ability of Pseudomonas aeruginosa to cause chronic infection is attributed to several virulence factors, biofilm formation, intrinsic and acquired resistance to many antibiotics. Anti-quorum sensing (QS) and anti-virulence therapy are promising alternatives to the existing antibiotic therapy. In this study, the effect of chitosan and the prepared chitosan-zinc oxide (CH/ZnO) nanocomposite on QS-dependent virulence factors and acyl homoserine lactone "AHL" production was studied. The chemical structure of the prepared CH/ZnO nanocomposite was characterized by FT-IR spectrum and XRD. The thermal stability and particle size were determined. Chitosan causes a significant decrease in AHL, biofilm, pyocyanin production and motility of P. aeruginosa. CH/ZnO nanocomposite augments the inhibitory activity of chitosan in both phenotypic and genotypic levels. Both chitosan and CH/ZnO nanocomposite downregulate the expression of LasI and RhlI genes using quantitative real-time PCR. The expression of RhlI gene in PAO1 is reduced by 1240 folds after treatment with CH/ZnO nanocomposite. The expression of LasI and RhlI genes in clinical isolates is reduced by 1778.07 and 627.29 folds upon treatment with CH/ZnO. These promising results may find a rescue in the battle of fighting P. aeruginosa by repressing its QS-dependent virulence factors.