The Biofilm Inhibition Properties of Glucosamine Gold Nanoparticles in Combination with Meropenem against Pseudomonas aeruginosa on the Endotracheal Tube: A Model of Biofilm-Related Ventilator-Associated Pneumonia.

Biofilm-related infections play a significant role in the development and persistence of ventilator-associated pneumonia. Pseudomonas aeruginosa (P. aeruginosa) frequently causes biofilm-related infections associated with ventilator tubing. Glucosamine gold nanoparticles (AuNPs) may exhibit antibiofilm properties; however, more studies, including combinatorial therapy with antibiotics, are needed to explore their potential applications in clinical settings. This study aims to investigate the biofilm inhibition properties of glucosamine AuNPs in combination with meropenem against P. aeruginosa ATCC 9027 on the endotracheal tube. A biofilm inhibition assay of glucosamine AuNPs at 0.02 mg/mL, both singly and in combination with meropenem at 1 mg/mL, was carried out against P. aeruginosa ATCC 9027 on an endotracheal tube using the tissue culture plate method. Scanning electron microscopy was performed for visualization. Glucosamine AuNPs at 0.02 mg/mL combined with meropenem at 1 mg/mL showed greater biofilm inhibition (72%) on the endotracheal tube than glucosamine nanoparticles at 0.02 mg/mL alone (26%) (p = 0.001). The scanning electron microscopic visualization revealed that the untreated P. aeruginosa biofilm was denser than the glucosamine nanoparticles-treated biofilm, whether combined with meropenem or using glucosamine nanoparticles alone. The combination of glucosamine AuNPs and meropenem may have the synergistic effect of inhibiting biofilm production of P. aeruginosa on the endotracheal tubes of patients with mechanical ventilation. Conducting additional experiments to explore the impact of combining glucosamine-coated gold nanoparticles (AuNPs) with meropenem on the inhibition of biofilm production by clinical P. aeruginosa isolates would be beneficial.

Evaluation of the microplastics in bivalves and water column at Pantai Teluk Likas, North Borneo, Malaysia.

Microplastics (MPs) are a pervasive pollutant in the marine environment. Pantai Teluk Likas in Sabah, Malaysia is one of the most visited beaches where tourism, recreational, and fisheries activities are high in this area. Hence, the area suffers from severe pollution, particularly from plastics. This study aims to quantify the microplastic composition in terms of color, shapes, and polymer types in marine bivalves (Anadara granosa, Glauconome virens, and Meretrix lyrata) and water column samples from Pantai Teluk Likas. All samples were digested using sodium hydroxide (NaOH) and incubated in the oven for at least 48 h. Serial filtration was done for each sample before they were observed under the dissecting microscope. The microplastics were identified and counted based on their physical attributes which were colors and shapes. The functional group of the polymers was determined using FTIR spectroscopy. Microplastics were found present in all samples collected. G. virens had the highest abundance of microplastics at 113.6 ± 6.5 particles/g followed by M. lyrata at 78.4 ± 3.7 particles/g. On the contrary, A. granosa had the least microplastics with an abundance of 24.4 ± 0.6 particles/g. Meanwhile, 110.0 ± 36.2 particles/L of microplastics were found in water column samples from Pantai Teluk Likas. Based on the analysis, fibers were the most common shape in bivalves, while fibers and films were common in the water column. In terms of colors, black, blue, and red were a few of the most abundant colors observed in both samples. The most common polymer detected in all bivalve species and water column samples is polycarbonate (PC), followed by polymethyl methacrylate (PMMA). Future study that focuses on the correlation between microplastic abundance in the marine biota and the water column is recommended to better understand microplastic availability and exposure.

In Vitro Assessment on Designing Novel Antibiofilms of Pseudomonas aeruginosa Using a Computational Approach.

An anti-biofilm that can inhibit the matrix of biofilm formation is necessary to prevent recurrent and chronic Pseudomonas aeruginosa infection. This study aimed to design compounds with a new mechanism through competitive inhibitory activity against phosphomannomutase/phosphoglucomutase (PMM/PGM), using in vitro assessment and a computational (in silico) approach. The active site of PMM/PGM was assessed through molecular redocking using L-tartaric acid as the native ligand and other small molecules, such as glucaric acid, D-sorbitol, and ascorbic acid. The docking program set the small molecules to the active site, showing a stable complex formation. Analysis of structural similarity, bioavailability, absorption, distribution, metabolism, excretion, and toxicity properties proved the potential application of ligands as an anti-biofilm. In vitro assessment with crystal violet showed that the ligands could reach up to 95.87% inhibition at different concentrations. The nitrocellulose membrane and scanning electron microscopic visualization showed that the untreated P. aeruginosa biofilm was denser than the ligand-treated biofilm.

An intestinal Candida albicans model for monomicrobial and polymicrobial biofilms and effects of hydrolases and the Bgl2 ligand.

Background and Aim: Candida albicans is the most prevalent human fungal pathogen. In biofilms, C. albicans becomes more resistant to antifungal agents because of the production of an extracellular matrix (ECM) that protects the yeast cells. This study aimed to determine the effects of hydrolase enzymes and the Bgl2 ligand on monomicrobial and polymicrobial biofilms. Materials and Methods: Biofilm induction in rats was carried out using streptomycin (25 mg/kg) and gentamicin (7.5 mg/kg) administered orally once per day for 5 days. Rats were injected subcutaneously with cortisone acetate (225 mg/kg) as an immunosuppressant on day 5. In addition, rats were orally administered C. albicans for the single microbial model and a combination of C. albicans with Escherichia coli for the polymicrobial model. Following the biofilm production, the groups were treated with glucosamine (8.57 mg/kg body weight) and Achatina fulica hydrolases (1.5 mL) orally for 2 weeks. The reduction of the biofilm was measured using confocal laser scanning microscopy (CLSM). Data were analyzed using a t-test, with a significance value of 95%. Results: CLSM images revealed a strong association between C. albicans and E. coli in the polymicrobial biofilm. On the contrary, the combination treatment using glucosamine and A. fulica hydrolases reduced the ECM of the single microbial biofilm (53.58%). However, treatment effectiveness against the matrix (19.17%) was reduced in the polymicrobial model. Conclusion: There is a strong association between C. albicans and E. coli in the formation of polymicrobial biofilms. The combination of glucosamine and the A. fulica enzyme can reduce the single microbial biofilm ECM; however, it is ineffective in the polymicrobial model.

A Novel 1,3-Β-Glucanase Gene from the Metagenomic Expression Library of Achatina Fulica's Digestive Gland.

1,3-ß-glucanase enzyme has been proved as antibiofilm by hydrolyzing the main component of extracellular matrix of C. albicans polymicrobial biofilm, to prevent resistancy during the use of antibiotics. The aim of this study is to construct a metagenomic expression library from Achatina fulica's digestive gland and to screen for a novel 1,3-ß-glucanase genes by using its specific substrate of laminarin. A cDNA expression library was constructed using the λTriplEx2 vector in the E. coli strain XL1-Blue. Cre-recombinase circularization was used to convert λTriplEx2 to pTriplEx2 in the E. coli strain BM 25.8;then IPTG induction was used to express 1,3-ß-glucanase. High-efficiency cDNA library of A. fulica's digestive gland was constructed, from where we obtained seventeen halo positive plaques, among them is a novel 1,3-ß-glucanase gene designated MkafGlu1. Its nucleotide sequence has similarities to the endo-1,3-ß-glucanase from Gossypium hirsutum, as well as the ß-glucanases from Paenibacillus mucilaginosus, Verticillium alfalfa, and Cryptopygus antarcticus of 45%, 40%, 38% and 37%, respectively. An open reading frame of 717 bp encoded a protein of 239 amino acids. A novel 1,3-ß-glucanase gene called MkafGlu1 was successfully expressed in E.coli BM 25.8 with activity of 1.07 U mL-1.

The Efficacy of Photodynamic Inactivation of the Diode Laser in Inactivation of the Candida albicans Biofilms With Exogenous Photosensitizer of Papaya Leaf Chlorophyll.

Introduction: Photodynamic inactivation has been developed to kill pathogenic microbes. In addition, some techniques have been introduced to minimize the biofilm resistance to antifungal properties in inhibiting cell growth. The principle of photodynamic inactivation different to antifungal drugs therapy which is resistant to biofilms. The presence of reactive oxygen species (ROS) that generating in photodynamic inactivation mechanisms can be damaging of biofilm cells and the principle of light transmission that could be penetrating in matrix layers of extracellular polymeric substance (EPS) until reaching the target cells at the base layers of biofilm. The present work aims to explore the potential of chlorophyll extract of papaya leaf as an exogenous photosensitizer to kill the Candida albicans biofilms after being activated by the laser. The potential of chlorophyll photosensitizer was evaluated based on the efficacy of inactivation C. albicans biofilm cell through a cell viability test and an organic compound test. Methods: The treatment of photoinactivation was administered to 12 groups of C. albicans biofilm for four days using the 445 nm laser and the 650 nm laser. The 445 nm and 650 nm lasers activated the chlorophyll extract of the papaya leaf (0.5 mg/L) at the same energy density. The energy density variation was determined as 5, 10, 20, 30 and 40 J/cm2 with the duration of exposure of each laser adjusted to the absorbance percentage of chlorophyll extract of the papaya leaf. Results: The absorbance percentage of chlorophyll extracts of the papaya leaf on wavelengths of 650 nm and 445 nm respectively were 22.26% and 60.29%, respectively. The most effective treated group was a group of the laser with the addition of chlorophyll, done by the 650 nm lasers with inactivation about 32% (P=0.001), while the 445 nm lasers only 25% (P=0.061). The maximum malondialdehyde levels by treatment of the laser 650 nm were (0.046±0.004) nmol/mg. Conclusion: The use of chlorophyll extract of the papaya leaf as a photosensitizer, resulted in the maximum spectrum of absorption at 414 nm and 668 nm, which produced a maximum reduction effect after photoinactivation up to 32% (with chlorophyll) and 25% (without chlorophyll). The utilization of chlorophyll extract of the papaya leaf would increase the antifungal effects with the activation by the diode laser in the biofilm of C. albicans.