Model For Gram-positive Bacteria Research Articles

Photostability and photodynamic antimicrobial profile of dye extracts from four (4) plants: prospects for eco-friendly low-cost food disinfection and topical biomedical applications.

In this study, photostability and photodynamic antimicrobial performance of dye extracts from Hibiscus sabdariffa (HS) calyces, Sorghum bicolor (SB) leaf sheaths, Lawsonia inermis (LI) leaves and Curcuma longa (CL) roots were investigated in Acetate-HCl (AH) Buffer (pH 4.6), Tris Base-HCl (TBH) Buffer (pH 8.6), distilled water (dH2O), and Phosphate Buffer Saline (PBS, pH 7.2) using Bacillus subtilis as model for gram positive bacteria, Escherichia coli as model for gram negative bacteria, phage MS2 as model for non-envelope viruses and phage phi6 as model for envelope viruses including SARS CoV-2 which is the causative agent of COVID-19. Our results showed that the photostability of the dye extracts is in the decreasing order of LI > CL > SB > HS. The dye extract-HS is photostable in dH2O but bleaches in buffers-AH, TBH and PBS. The rate of bleaching is higher in AH compared to in TBH and PBS. The bleaching and buffers affected the photodynamic and non-photodynamic antimicrobial activity of the dye extracts. The photodynamic antibacterial activity of the dye extracts is in the decreasing order of CL > HS > LI > SB while the non-photodynamic antibacterial activity is in the decreasing order of LI > CL > HS > SB. The non-photodynamic antiviral activity pattern observed is the same as that of non-photodynamic antibacterial activity observed. However, the photodynamic antiviral activity of the dye extracts is in the decreasing order of CL > LI > HS > SB. Given their performance, the dye extracts maybe mostly suitable for environmental applications including fresh produce and food disinfection, sanitation of hands and contact surfaces where water can serve as diluent for the extracts and the microenvironment is free of salts.

Vancomycin-Loaded Furriness Amino Magnetic Nanospheres for Rapid Detection of Gram-Positive Water Bacterial Contamination.

Bacterial pathogens pose high threat to public health worldwide. Different types of nanomaterials have been synthesized for the rapid detection and elimination of pathogens from environmental samples. However, the selectivity of these materials remains challenging, because target bacterial pathogens commonly exist in complex samples at ultralow concentrations. In this study, we fabricated novel furry amino magnetic poly-L-ornithine (PLO)/amine-poly(ethylene glycol) (PEG)-COOH/vancomycin (VCM) (AM-PPV) nanospheres with high-loading VCM for vehicle tracking and the highly efficient capture of pathogens. The magnetic core was coated with organosilica and functionalized with cilia. The core consisted of PEG/PLO loaded with VCM conjugated to Gram-positive bacterial cell membranes, forming hydrogen bonds with terminal peptides. The characterization of AM-PPV nanospheres revealed an average particle size of 56 nm. The field-emission scanning electron microscopy (FE-SEM) micrographs showed well-controlled spherical AM-PPV nanospheres with an average size of 56 nm. The nanospheres were relatively rough and contained an additional 12.4 nm hydrodynamic layer of PLO/PEG/VCM, which provided additional stability in the suspension. The furry AM-PPV nanospheres exhibited a significant capture efficiency (>90%) and a high selectivity for detecting Bacillus cereus (employed as a model for Gram-positive bacteria) within 15 min, even in the presence of other biocompatible pathogens. Moreover, AM-PPV nanospheres rapidly and accurately detected B. cereus at levels less than 10 CFU/mL. The furry nano-design can potentially satisfy the increasing demand for the rapid and sensitive detection of pathogens in clinical and environmental samples.

Exploring the potential of highly charged Ru(II)- and heteronuclear Ru(II)/Cu(II)-polypyridyl complexes as antimicrobial agents

The antimicrobial potential of two ruthenium(II) polypyridyl complexes, [Ru(phen)2L1]2+ and [Ru(phen)2L2]2+ (phen = 1,10-phenanthroline) containing the 4,4′-(2,5,8,11,14-pentaaza[15])-2,2’-bipyridilophane (L1) and the 4,4′-bis-[methylen-(1,4,7,10-tetraazacyclododecane)]-2,2′ bipyridine (L2) units, is herein investigated. These peculiar polyamine frameworks afford the formation of highly charged species in solution, influence the DNA-binding and cleavage properties of compounds, but they do not undermine their singlet oxygen sensitizing capacities, thus making these complexes attractive 1O2 generators in aqueous solution. L1 and L2 also permit to stably host Fenton -active Cu2+ ion/s, leading to the formation of mixed Ru2+/Cu2+ forms capable to further strengthen the oxidative damages to biological targets. Herein, following a characterization of the Cu2+ binding ability by [Ru(phen)2L2]2+, the water-octanol distribution coefficients, the DNA binding, cleavage and 1O2 sensitizing properties of [Ru(phen)2L2]2+ and [Cu2Ru(phen)2L2]6+ were analysed and compared with those of [Ru(phen)2L1]2+ and [CuRu(phen)2L1]4+. The antimicrobial activity of all compounds was evaluated against B. subtilis, chosen as a model for gram-positive bacteria, both under dark and upon light-activation. Our results unveil a notable phototoxicity of [Ru(phen)2L2]2+ and [Cu2Ru(phen)2L2]6+, with MIC (minimal inhibitory concentrations) values of 3.12 μM. This study highlights that the structural characteristics of polyamine ligands gathered on highly charged Ru(II)-polypyridyl complexes are versatile tools that can be exploited to achieve enhanced antibacterial strategies.

Establishment of a Markerless Mutation Delivery System in Bacillus subtilis Stimulated by a Double-Strand Break in the Chromosome

Bacillus subtilis has been a model for gram-positive bacteria and it has long been exploited for industrial and biotechnological applications. However, the availability of facile genetic tools for physiological analysis has generally lagged substantially behind traditional genetic models such as Escherichia coli and Saccharomyces cerevisiae. In this work, we have developed an efficient, precise and scarless method for rapid multiple genetic modifications without altering the chromosome of B. subtilis. This method employs upp gene as a counter-selectable marker, double-strand break (DSB) repair caused by exogenous endonuclease I-SceI and comK overexpression for fast preparation of competent cell. Foreign dsDNA can be simply and efficiently integrated into the chromosome by double-crossover homologous recombination. The DSB repair is a potent inducement for stimulating the second intramolecular homologous recombination, which not only enhances the frequency of resolution by one to two orders of magnitude, but also selects for the resolved product. This method has been successfully and reiteratively used in B. subtilis to deliver point mutations, to generate in-frame deletions, and to construct large-scale deletions. Experimental results proved that it allowed repeated use of the selectable marker gene for multiple modifications and could be a useful technique for B. subtilis.

The importance of bacterial membrane composition in the structure and function of aurein 2.2 and selected variants

For cationic antimicrobial peptides to become useful therapeutic agents, it is important to understand their mechanism of action. To obtain high resolution data, this involves studying the structure and membrane interaction of these peptides in tractable model bacterial membranes rather than directly utilizing more complex bacterial surfaces. A number of lipid mixtures have been used as bacterial mimetics, including a range of lipid headgroups, and different ratios of neutral to negatively charged headgroups. Here we examine how the structure and membrane interaction of aurein 2.2 and some of its variants depend on the choice of lipids, and how these models correlate with activity data in intact bacteria (MICs, membrane depolarization). Specifically, we investigated the structure and membrane interaction of aurein 2.2 and aurein 2.3 in 1:1 cardiolipin/1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (CL/POPG) (mol/mol), as an alternative to 1:1 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine(POPC)/POPG and a potential model for Gram positive bacteria such as S. aureus. The structure and membrane interaction of aurein 2.2, aurein 2.3, and five variants of aurein 2.2 were also investigated in 1:1 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE)/POPG (mol/mol) lipids as a possible model for other Gram positive bacteria, such as Bacillus cereus. Solution circular dichroism (CD) results demonstrated that the aurein peptides adopted α-helical structure in all lipid membranes examined, but demonstrated a greater helical content in the presence of POPE/POPG membranes. Oriented CD and 31P NMR results showed that the aurein peptides had similar membrane insertion profiles and headgroup disordering effects on POPC/POPG and CL/POPG bilayers, but demonstrated reduced membrane insertion and decreased headgroup disordering on mixing with POPE/POPG bilayers at low peptide concentrations. Since the aurein peptides behaved very differently in POPE/POPG membrane, minimal inhibitory concentrations (MICs) of the aurein peptides in B. cereus strain C737 were determined. The MIC results indicated that all aurein peptides are significantly less active against B. cereus than against S. aureus and S. epidermidis. Overall, the data suggest that it is important to use a relevant model for bacterial membranes to gain insight into the mode of action of a given antimicrobial peptide in specific bacteria.

Microbial inactivation using plasma-activated water obtained by gliding electric discharges

To evaluate the microbial disinfection efficacy of a plasmachemical solution obtained by the activation of water with gliding electric discharges. Distilled water was activated for 5 min by a nonthermal quenched plasma of the glidarc type operating in humid air and at atmospheric pressure. The plasma-activated water (PAW) was then used to treat planktonic and adherent cells of Staphylococcus epidermidis, Leuconostoc mesenteroides (as models of Gram-positive bacteria), Hafnia alvei (a Gram-negative bacteria) and Saccharomyces cerevisiae (as a yeast model). The treatments were less efficient on adherent cells than on planktonic cells in the case of bacteria, but not of S. cerevisiae. Inactivation was more effective for bacteria than for the yeast. Significant reductions in microbial populations were achieved in all cases, demonstrating the effectiveness of this new approach to treat contaminated media. PAW is a promising solution with potential application to the decontamination of equipment and surfaces.

Antimicrobial Characteristics of Silver Aerosol Nanoparticles againstBacillus subtilisBioaerosols

Silver aerosol nanoparticles generated by an atomizer were investigated as an antimicrobial agent against B. subtilis bioaerosols, a model for Gram-positive bacteria. The bioaerosols and silver aerosol nanoparticles were mixed in a test duct for 15 s and sampled on nutrient agar plates by an impactor. Peak concentrations of the silver aerosol nanoparticles at the mobility diameter of 80 nm were controlled from 1.5 × 105 to 1.0 × 106 particles/cm3, depending on the concentration of silver hydrosol nanoparticles. The maximal antimicrobial efficiency of silver aerosol nanoparticles was 76% under selected experimental conditions.