Isolation and characterisation of a novel Silviavirus bacteriophage promising antimicrobial agent against methicillin-resistant Staphylococcus aureus infections.

The increasing prevalence of methicillin-resistant Staphylococcus aureus (MRSA) emphasises the urgent need for novel antimicrobial agents as alternatives to antibiotics. Bacteriophage therapy is one of the most promising antimicrobial strategies. Here, we isolated and comprehensively characterized a novel Staphylococcus phage, vB_SauM_VL10 (VL10), from urban sewage. The VL10 genome displays 141,746 bp of linear double-stranded DNA, containing 193 open reading frames and lacking tRNA, virulence, or antibiotic resistance genes. Phylogenetic analysis categorizes VL10 as a novel species within the Silviavirus genus, Twortvirinae subfamily. VL10 exhibits lytic behaviour characterized by efficient adsorption, a short latent period, and substantial burst size, with environmental stability. It demonstrates lytic activity against 79.06% of tested S. aureus strains, highlighting its species specificity. Additionally, VL10 effectively targets MRSA biofilms, reducing biomass and viable cells. In MRSA-infected G. mellonella larvae, VL10 enhances survival rates, supporting its potential for phage therapy applications. Moreover, the emergence of VL10-resistant S. aureus strains associated with fitness trade-offs, including reduced growth, biofilm formation, and virulence. Altogether, these findings emphasize VL10 as a promising candidate for developing therapeutic agents against MRSA infections, providing insights into phage biology and resistance dynamics.

Enhanced Antibacterial Activity at Ag-Cu Nanojunctions: Unveiling the Mechanism with Simple Surfaces of CuNPs-on-Ag Films.

Deposition of CuNPs on silver film gives rise to the formation of active Ag-Cu interfaces leading to dramatic enhancements in antibacterial activity against Escherichia coli. Transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDAX) analyses reveal that CuNPs are covered in a thin Cu2O shell, while X-ray photoelectron spectroscopy measurements (XPS) reveal that the Ag film samples contain significant amounts of Ag2O. XPS analyses show that the deposition of CuNPs on Ag films leads to the formation of a photoactive Ag2O-Cu2O heterostructure. Following a Z-scheme mechanism, electrons from the conduction band of Ag2O recombine with photogenerated holes from the valence band of Cu2O. Consequently, electrons at Cu2O's conduction band render Cu reduced and cause reductive activation of surface oxygen species on Cu forming reactive oxygen species (ROS). Interaction between metallic Cu and ROS species leads to the formation of a Cu(OH)2 phase. Both ROS and Cu(OH)2 species have previously been reported to lead to enhanced antibacterial properties. Holes on Ag2O produce a highly oxidized AgO phase, a phase reported to exhibit excellent antibacterial properties. Quantitative analysis of Cu and Ag high-resolution X-ray photoelectron spectroscopy (HR-XPS) spectra directly reveals several-fold increases in these active phases in full agreement with the observed increase in antibacterial activities. This study provides insight and surface design parameters by elucidating the important roles of Ag and Cu's bifunctionality as active antibacterial materials.

Supramolecular self-assembled polymeric nanospheres based on hydrazino naphthalimide functionalised pillar[5]arene for long chain aldehyde detection.

A novel fluorescent sensor HNP5A is constructed consisting of bis-hydrazine naphthalimide decorated pillar[5]arene. Interestingly, this sensor possessed the potential for sensitive and selective detection of long-chain aldehydes, particularly nonanal (C9), and subsequently formed supramolecular pseudorotaxane polymeric nanoparticles inducing a strong fluorescence enhancement. In addition, this as-produced HNP5A⊂C9 exhibited an unexpected reduction of Ag+ to produce AgNPs in an aqueous system and the resulting AgNPs-HNP5A⊂C9 demonstrated a significant fluorescence enhancement under metal-enhanced fluorescence (MEF) behaviour.

Osmotic Pressure as Driving Force for Reducing the Size of Nanoparticles in Emulsions.

We describe here a method to decrease particle size of nanoparticles synthesized by miniemulsion polymerization. Small nanoparticles or nanocapsules were obtained by generating an osmotic pressure to induce the diffusion of monomer molecules from the dispersed phase of a miniemulsion before polymerization to an upper oil layer. The size reduction is dependent on the difference in concentration of monomer in the dispersed phase and in the upper oil layer and on the solubility of the monomer in water. By labeling the emulsion droplets with a copolymer of stearyl methacrylate and a polymerizable dye, we demonstrated that the migration of the monomer to the upper hexadecane layer relied on molecular diffusion rather than diffusion of monomer droplets to the oil layer. Moreover, surface tension measurements confirmed that the emulsions were still in the miniemulsion regime and not in the microemulsion regime. The particle size can be tuned by controlling the duration during which the miniemulsion stayed in contact with the hexadecane layer, the interfacial area between the miniemulsion and the hexadecane layer and by the concentration of surfactant. Our method was applied to reduce the size of polystyrene and poly(methyl methacrylate) nanoparticles, nanocapsules of a copolymer of styrene and methyl methacrylic acid, and silica nanocapsules. This work demonstrated that a successful reduction of nanoparticle size in the miniemulsion process can be achieved without using excess amounts of surfactant. The method relies on building osmotic pressure in oil droplets dispersed in water which acts as semipermeable membrane.

A novel virulent Litunavirus phage possesses therapeutic value against multidrug resistant Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a notable nosocomial pathogen that can cause severe infections in humans and animals. The emergence of multidrug resistant (MDR) P. aeruginosa has motivated the development of phages to treat the infections. In this study, a novel Pseudomonas phage, vB_PaeS_VL1 (VL1), was isolated from urban sewage. Phylogenetic analyses revealed that VL1 is a novel species in the genus Litunavirus of subfamily Migulavirinae. The VL1 is a virulent phage as no genes encoding lysogeny, toxins or antibiotic resistance were identified. The therapeutic potential of phage VL1 was investigated and revealed that approximately 56% (34/60 strains) of MDR P. aeruginosa strains, isolated from companion animal diseases, could be lysed by VL1. In contrast, VL1 did not lyse other Gram-negative and Gram-positive bacteria suggesting its specificity of infection. Phage VL1 demonstrated high efficiency to reduce bacterial load (~ 6 log cell number reduction) and ~ 75% reduction of biofilm in pre-formed biofilms of MDR P. aeruginosa. The result of two of the three MDR P. aeruginosa infected Galleria mellonella larvae showed that VL1 could significantly increase the survival rate of infected larvae. Taken together, phage VL1 has genetic and biological properties that make it a potential candidate for phage therapy against P. aeruginosa infections.

A new species of Chlamydia isolated from Siamese crocodiles (Crocodylus siamensis).

Chlamydia is a known pathogen in both saltwater and freshwater crocodiles. However, the exact species/strain has not been clearly identified. In this study, we successfully cultivated Siamese crocodile Chlamydia in McCoy cells at a temperature of 30°C. Electron microscopy; phylogeny based on nine conserved taxonomically informative markers, on ompA, or on seven housekeeping genes; and whole-genome sequencing and analysis of the isolate confirmed the identity of the isolate as a new member of the genus Chlamydia, a new species that we name Chlamydia crocodili.

Effect of Pore Connectivity of Pore-Opened Hierarchical MOR Zeolites on Catalytic Behaviors and Coke Formation in Ethanol Dehydration.

The hierarchical zeolite is one of the most promising materials for catalytic applications. However, the effect of its pore connectivity on catalytic behaviors and coke formation has not clearly been revealed. In this contribution, we demonstrate the visualization of the mesopore architecture in three-dimensional perspectives together with the pore connectivity network of pore-opened hierarchical mordenite (MOR), fabricated by the seed-assisted template-free synthesis followed by the fluoride treatment via the electron tomography (ET) technique. Interestingly, the pore-opened zeolites clearly display higher catalytic performance (approximately 80% of ethylene yield) in ethanol dehydration with respect to the parent one due to their additional pore-opened structures connected to the external surfaces of zeolites. In addition, the effect of pore connectivity network on the coke location and type obtained from ethanol conversion has been observed. It was found that the porous structure of the etched sample is directly connected to the external surface, and then, the large area of crystals can contribute to the reaction. Conversely, only a small amount of closed mesopores is observed inside the crystals in the case of the untreated sample, and therefore, the molecules cannot easily penetrate inside crystals for the catalytic reaction. These results open up promising perspectives for the development of hierarchical catalysts including fabrication by the template-free synthesis approach, pore-architecture characterization, and catalytic applications.

ANTI-AMEBIC ACTIVITY OF DIOSGENIN ON NAEGLERIA FOWLERI TROPHOZOITES.

The aim of this study was to investigate the activity of diosgenin against Naegleria fowleri trophozoites at the cellular and molecular levels. Diosgenin (100 µg/ml; 241.2 µM) had a 100% inhibitory effect on N. fowleri trophozoites (5 x 10(5) cell/ml). Scanning electron micrograph revealed diosgenin decreased the number of sucker-like apparatuses and food cup formation among N. fowleri trophozoites at 3 and 6 hours post-exposure, respectively. Diosgenin down-regulated the nf cysteine protease gene expression of N. fowleri trophozoites at 6 and 12 hours post-exposure. The toxicity to mammalian cells caused by diosgenin at therapeutic dose was less than amphotericin B, the current drug used to treat N. fowleri infections. Our findings suggest diosgenin has activity against the surface membrane and the nf cysteine pro tease of N. fowleri trophozoites. However, the other mechanisms of action of diosgenin against N. fowleri trophozoites require further exploration.

Effect of synthetic antimicrobial peptides on Naegleria fowleri trophozoites.

We evaluated the effect of tritrpticin, lactoferrin, killer decapeptide and scrambled peptide in vitro against Naegleria fowleri trophozoites compared with amphotericin B. Tritrpticin (100 microg/ml) caused apoptosis of N. fowleri trophozoites (2x10(5) cells/ml), while lactoferrin, killer decapeptide and scrambled peptide did not. On Gormori trichrome staining, tritrpticin affected the elasticity of the surface membrane and reduced the size of the nuclei of N. fowleri trophozoites. The ultrastructure surface membrane and food cup formation of the trophozoites were 100% inhibited. These results are consistent with inhibition of the nfa1, Mp2CL5 of the treated trophozoite, which plays a role in food cup formation. Tritrpticin 100 microg/ml was not toxic against SK-N-MC cells. Our findings suggest tritrpticin has activity against the surface membrane and nfa1 and Mp2CL5 of N. fowleri trophozoites and could be developed as a potential therapeutic agent.