2D TiO2 nanosheets decorated via sphere-like BiVO4: a promising non-toxic material for liquid phase photocatalysis and bacterial eradication.

An in-depth investigation was conducted on a promising composite material (BiVO4/TiO2), focusing on its potential toxicity, photoinduced catalytic properties, as well as its antibiofilm and antimicrobial functionalities. The preparation process involved the synthesis of 2D-TiO2 using the lyophilization method, which was subsequently functionalized with sphere-like BiVO4. Finally, we developed BiVO4/TiO2 S-scheme heterojunctions which can greatly promote the separation of electron-hole pairs to achieve high photocatalytic performance. The evaluation of concentration- and time-dependent viability inhibition was performed on human lung carcinoma epithelial A549 cells. This assessment included the estimation of glutathione levels and mitochondrial dehydrogenase activity. Significantly, the BiVO4/TiO2 composite demonstrated minimal toxicity towards A549 cells. Impressively, the BiVO4/TiO2 composite exhibited notable photocatalytic performance in the degradation of rhodamine B (k =0.135 min-1) and phenol (k = 0.016 min-1). In terms of photoinduced antimicrobial performance, the composite effectively inactivated both gram-negative E. coli and gram-positive E. faecalis bacteria upon 60-min of UV-A light exposure, resulting in a significant log6(log10CFU/mL) reduction in bacterial count. These promising results can be attributed to the unique 2D morphology of TiO2 modified by sphere-like BiVO4, leading to an increased generation of (intracellular)hydroxyl radicals, which plays a crucial role in treatments of both organic pollutants and bacteria.

Mechanism of Action and Efficiency of Ag3PO4-Based Photocatalysts for the Control of Hazardous Gram-Positive Pathogens.

Silver phosphate and its composites have been attracting extensive interest as photocatalysts potentially effective against pathogenic microorganisms. The purpose of the present study was to investigate the mechanism of bactericidal action on cells of opportunistic pathogens. The Ag3PO4/P25 (AGP/P25) and Ag3PO4/HA (HA/AGP) powders were prepared via a co-precipitation method. Thereafter, their antimicrobial properties against Enterococcus faecalis, Staphylococcus epidermidis, and Staphylococcus aureus (clinical and reference strains) were analyzed in the dark and after exposure to visible light (VIS). The mechanism leading to cell death was investigated by the leakage of metabolites and potassium ions, oxidative stress, and ROS production. Morphological changes of the bacterial cells were visualized by transmission electron microscopy (TEM) and scanning transmission electron microscopy with energy-dispersive X-ray spectroscopy (SEM EDS) analysis. It has been shown that Ag3PO4-based composites are highly effective agents that can eradicate 100% of bacterial populations during the 60 min photocatalytic inactivation. Their action is mainly due to the production of hydroxyl radicals and photogenerated holes which lead to oxidative stress in cells. The strong affinity to the bacterial cell wall, as well as the well-known biocidal properties of silver itself, increase undoubtedly the antimicrobial potential of the Ag3PO4-based composites.

Biocidal activity of multifunctional cuprite-doped anion exchanger - Influence of bacteria type and medium composition.

The study presents unconventional, bifunctional, heterogeneous antimicrobial agents - Cu2O-loaded anion exchangers. The synergetic effect of a cuprous oxide deposit and polymeric support with trimethyl ammonium groups was studied against the reference strains of Enterococcus faecalis ATCC 29212 and Pseudomonas aeruginosa ATCC 27853. Biological testing (minimum bactericidal concentration, MBC), time- and dose-dependent bactericidal effect (under different conditions - medium composition and static/dynamic culture) demonstrated promising antimicrobial activity and confirmed its multimode character. The standard values of MBC, for all studied hybrid polymers and bacteria, were similar (64-128 mg/mL). However, depending on the medium conditions, due to the copper release into the bulk solution, bacteria were actively killed even at much lower doses of the hybrid polymer (25 mg/mL) and low Cu(II) concentrations in solution (0.01 mg/L). Simultaneously, confocal microscopic studies confirmed the effective inhibition of bacterial adhesion and biofilm formation on their surface. The studies conducted under different conditions showed also the influence of the structure and physical properties of studied materials on the biocidal efficacy and an antimicrobial action mechanism was proposed that could be significantly affected by electrostatic interactions and copper release to the solution. Although the antibacterial activity was also dependent on various strategies of bacterial cell resistance to heavy metals present in the aqueous medium, the studied hybrid polymers are versatile and efficient biocidal agents against bacteria of both types, Gram-positive and Gram-negative. Therefore, they can be a convenient alternative for point-of-use water disinfection systems providing water quality in medical devices such as dental units, spa equipment, and aesthetic devices used in the cosmetic sector.

Photo-responsive functional gold nanocapsules for inactivation of community-acquired, highly virulent, multidrug-resistant MRSA.

The indiscriminate and sporadic use of antibiotics has contributed to the emergence of drug resistance phenomenon in bacteria including but not limited to Staphylococcus aureus. These drug-resistant bacteria have been threatening safety in hospitals and adversely affecting human health. Here we report a strategy to design photo-stimulated theranostic nanoprobes against methicillin-resistant Staphylococcus aureus (MRSA) "superbug" USA300. The nanocapsule probe is based on gold nanorods (GNRs) coated with pegylated thiol, mPEG-SH, which has been further modified by adding successively a natural antibacterial compound such as curcumin, and a cell targeting deoxyribonucleic acid (DNA) aptamer. We have used this novel gold nanocapsules for near-infrared (NIR) photophysical stimulation against pathogenic bacteria. We have found that the novel nanocapsule blocks biofilm formation and kills bacteria by photothermal action that causes disruption of the bacterial cell wall and membrane. In this approach, multiple drug-resistant Staphylococcus aureus has been captured by these nanocapsules through DNA aptamer targeting. All of the trapped bacteria could be killed in 30 minutes during the NIR stimulation due to the combination of photothermal effect, the generation of reactive oxygen species (ROS) and a loss of transmembrane potential (Δψ). Importantly we did not notice any resistance developed against the photothermal treatment. This is remarkable from an anti-biofilm activity point of view. Importantly, these multifunctional nanocapsules have also shown a surface enhanced Raman spectroscopy (SERS) effect, which could be used to evaluate the success of the inactivation effect during treatment. These results indicate that nanocapsule-based photo treatment can be an alternative antibacterial strategy without contributing to antibiotic resistance, and thus can be used for both environmental and therapeutic applications.

In Silico Screening and In Vitro Assessment of Natural Products with Anti-Virulence Activity against Helicobacter pylori.

Helicobacter pylori is one of the most frequent human pathogens and a leading etiological agent of various gastric diseases. As stringent response, coordinated by a SpoT protein, seems to be crucial for the survivability of H. pylori, the main goal of this article was to use in silico computational studies to find phytochemical compounds capable of binding to the active site of SpoT from H. pylori and confirm the ability of the most active candidates to interfere with the virulence of this bacterium through in vitro experiments. From 791 natural substances submitted for the virtual screening procedure, 10 were chosen and followed for further in vitro examinations. Among these, dioscin showed the most interesting parameters (the lowest MIC, the highest anti-biofilm activity in static conditions, and a relatively low stimulation of morphological transition into coccoids). Therefore, in the last part, we extended the research with a number of further experiments and observed the ability of dioscin to significantly reduce the formation of H. pylori biofilm under Bioflux-generated flow conditions and its capacity for additive enhancement of the antibacterial activity of all three commonly used antibiotics (clarithromycin, metronidazole, and levofloxacin). Based on these results, we suggest that dioscin may be an interesting candidate for new therapies targeting H. pylori survivability and virulence.

Antimicrobial O-Alkyl Derivatives of Naringenin and Their Oximes Against Multidrug-Resistant Bacteria.

New antimicrobial agents are needed to address infections caused by multidrug-resistant bacteria. Here, we are reporting novel O-alkyl derivatives of naringenin and their oximes, including novel compounds with a naringenin core and O-hexyl chains, showing activity against clinical strains of clarithromycin-resistant Helicobacter pylori, vancomycin-resistant Enterococcus faecalis, methicillin-resistant Staphylococcus aureus, and beta-lactam-resistant Acinetobacter baumannii and Klebsiella pneumoniae. The minimum inhibitory concentrations (MICs), which provide a quantitative measure of antimicrobial activity, were in the low microgram range for the selected compounds. Checkerboard assays for the most active compounds in combination with antibiotics revealed interactions that varied from synergistic to neutral.

Photo-catalytic inactivation of an Enterococcus biofilm: the anti-microbial effect of sulphated and europium-doped titanium dioxide nanopowders.

The control and prevention of biofilm-related infections is an important public healthcare issue. Given the increasing antibiotic resistance among bacteria and fungi that cause serious infections in humans, promotion of new strategies combating microorganisms has been essential. One attractive approach to inactivate microorganisms is the use of semiconductor photo-catalysis, which has become the subject of extensive research. In this study, the bactericidal properties of four photo-catalysts, TiO2, TiO2-S, TiO2-Eu and TiO2-Eu-S, were investigated against established 24, 48, 72 and 96 h biofilms of Enterococcus The exposure of biofilms to the catalysts induced the production of superoxide radical anions. The best photo-catalytic inactivation was achieved with the TiO2-Eu-S and TiO2-S nanopowders and 24 h biofilms. Transmission electron microscopy images showed significant changes in the structure of the biofilm cells following photo-inactivation. The results suggest that doping with europium and modifying the surface with sulphate groups enhanced the bactericidal activity of the TiO2 nanoparticles against enterococcal biofilms.

Activity of fluconazole and its Cu(II) complex towards Candida species.

Candida species, although they are present as commensal organisms in the digestive tract of healthy individuals, can produce a broad spectrum of serious illnesses in compromised hosts. Fluconazole, a water-soluble triazole with bioavailability greater than 90 %, has been extensively used to treat a wide range of Candida infections. However, a growing resistance of microorganisms in the treatment leads to the discovery of new drugs or modifications of existing ones. The aim of the present study was to investigate whether coordination of Cu(II) ions to fluconazole affects its antifungal activity. The in vitro susceptibility tests and antifungal studies were performed with two Candida spp.: Candidaglabrata and Candida albicans. Overall, 34 strains of the former and 16 strains of the latter were treated with fluconazole, its Cu(II) complex and free Cu(II) ions. The obtained MIC values in 16 cases of the C. glabrata and in 5 cases of the C. albicans were lower for the complex in comparison to the drug. This implies that the complex is more effective against particular strains than the parent drug. The most significant improvement in the complex drug efficacy was observed for fluconazole-resistant species.

The influence of biodegradable gemini surfactants, N,N'-bis(1-decyloxy-1-oxopronan-2-yl)-N,N,N',N' tetramethylpropane-1,3-diammonium dibromide and N,N'-bis(1-dodecyloxy-1-oxopronan-2-yl) N,N,N',N'-tetramethylethane-1,2-diammonium dibromide, on fungal biofilm and adhesion.

A group of biodegradable alanine-derived gemini quaternary ammonium salts (bromides and chlorides) with various alkyl chains and spacer lengths was tested for anti-adhesive and anti-biofilm activity. The strongest antifungal activity was exhibited by bromides with 10 and 12 carbon atoms within hydrophobic chains (N,N'-bis(1-decyloxy-1-oxopronan-2-yl)-N,N,N',N'-tetramethylpropane-1,3-diammonium dibromide and N,N'-bis(1-dodecyloxy-1-oxopronan-2-yl)-N,N,N',N'-tetramethylethane-1,2-diammonium dibromide). It was also demonstrated that these gemini surfactants enhanced the sensitivity of Candida albicans to azoles (itraconazole and fluconazole) and polyenes (amphotericin B and nystatine). Gemini quaternary ammonium salts effectively inhibited fungal cell adhesion to polystyrene and silicone surface. These compounds reduced C. albicans filamentation and eradicated C. albicans and Rhodotorula mucilaginosa biofilms, as it was shown in crystal violet and fluorescent staining. None of the tested compounds were cytotoxic against yeast mitochondrial metabolism.

Enterococcus--virulence and susceptibility to photodynamic therapy of clinical isolates from Lower Silesia, Poland.

BACKGROUND: A collection of 400 enterococcal isolates from clinical samples of hospitalized patients were studied for their virulence traits according to the isolation site. Formation of biofilm and production of DNase, hemolysin, lipase, and gelatinase were characterized. Biofilms of selected strains were examined for their susceptibility to antimicrobial photodynamic therapy (aPDT). METHODS: All strains were tested for biofilm production by microtiter method and the activity of hemolysin, gelatinase, lipase, and DNase by plate method with an adequate substrate. Photodynamic therapy with Photolon and red laser light was performed towards 48 h biofilms of eight representative strains. The viability of biofilms was tested by the BactLight assay and visualized under a fluorescent microscope. RESULTS: Among the 400 isolates, 69.8% strains of Enterococcus faecalis, 30% of Enterococcus faecium, and 0.2% of Enterococcus casseliflavus were identified. In vitro, production of biofilm was found in 65.7% of enterococci. Biofilm-positive strains were isolated from urinary tract infections (81%), wound infections (71%), respiratory tract infections (62%), and gastrointestinal tract (47%) (colonization). Hemolysin activity was observed in 28.5%, gelatinase in 24.5%, lipase in 23%, and DNase in 3.5% of all, mostly biofilm-positive, isolates. Photodynamic therapy with Photolon efficiently reduced the enterococcal biofilms. CONCLUSIONS: The study demonstrated the high prevalence of biofilm-producing clinical enterococci, their virulence potential being higher than for biofilm-negative strains, and the susceptibility to aPDT of mature biofilms produced by strains, regardless of their species and site of isolation.

Antibacterial activity of gemini quaternary ammonium salts.

A series of gemini quaternary ammonium salts (chlorides and bromides), with various hydrocarbon chain and spacer lengths, were tested. These compounds exhibited antibacterial activity against both Gram-positive and Gram-negative bacteria and were not mutagenic. The strongest antibacterial effect was observed for TMPG-10 Cl (against Pseudomonas aeruginosa ATCC 27853) and TMPG-12 Br (against Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 11229 and clinical ESBL(+) isolate 434) surfactants. These compounds inhibited the adhesion of Staphylococcus epidermidis ATCC 35984 to a polystyrene surface and eradicated biofilm formed by P. aeruginosa PAO1. The activity of studied compounds was dependent on hydrocarbon chain length.

Enterococcus in wound infections: virulence and antimicrobial resistance.

Enterococci, a complex group of facultative pathogens have become increasingly isolated in various hospital settings. They are considerable frequently cultured from traumatic and surgical wounds. We investigated 57 strains of the species E. faecalis, E. faecium and E. casseliflavus isolated from infected wounds. Their ability to produce virulence factors and their sensitivity to antibiotics were evaluated using phenotypic and genotyping methods. In the phenotype studies, significant portion of the isolates produced biofilm (66.7%) and gelatinase (36.8%). Nearly 30% of the strains expressed hemolytic properties. Only a few produced DNAse (15.8%) and lipase (7.0%). The genes esp, gelE, cylA, cylB, cylM and agg were detected in most of the isolates (38.6-87.7%). All the isolated enterococci were susceptible to vancomycin and were characterized by their low resistance to antibiotics, except aminoglycosides (HLR).

In situ photoexcitation of silver-doped titania nanopowders for activity against bacteria and yeasts.

Photocatalytic and in situ microbial activity of the amorphous and annealed states of Ag-doped and un-doped titania were examined. Studies on their structure, morphology, composition, and the photo-absorption characteristics of these materials were performed. These results were correlated with the photocatalytic and microbial activity against methicillin resistant Staphylococcus aureus K324 (MRSA), methicillin susceptible S. aureus ATCC 25923 (MSSA), Escherichia coli PA 170, and yeasts Candida albicans ATCC 90028. The annealed powders containing anatase form of titania exhibited relatively higher photocatalytic activity,corresponding to activity against MRSA,when exposed to UV-A radiation. In comparison, amorphous powders exhibited low photoactivity and showed poor antibacterial performance against MRSA under UV-A exposure. Doping of amorphous titania with Ag resulted in an anti-MRSA effect without exposure to UV radiation. In the Ag-doped crystalline anatase samples, the size of Ag primary nanocrystallites increased, which led to the decrease in the surface concentration of Ag and detriment anti-MRSA activity.

Virulence of Enterococcus isolates collected in Lower Silesia (Poland).

148 enterococcal strains: E.faecalis (108), E.faecium (35), E.gallinarum (3), E.casseliflavus (1) and E.durans (1) from various clinical specimens were investigated for their ability to adhere to Caco-2 and HEp-2 cell lines, and also for the presence of the esp gene, biofilm formation, production of haemolysins, DNAse and lipase. Several types of enterococcal adhesion to both cell lines were noted. An aggregative adherence was the most frequent among E.faecalis and E.faecium isolates. Other species presented various adhesive types. The occurrence of virulence factors in the whole group of strains was as follows: esp gene in 53.4%, biofilm in 45.3%, haemolysins in 15.5%, DNAse in 12.2% and lipase in 33.1% of enterococcal isolates. It appears that the adherence of the enterococci studied was not significantly associated with the presence of virulence factors.

The carrier state of shiga-like toxin II (SLT II) and hemolysin-producing enteroaggregative Escherichia coli strain.

Shiga-like toxin-producing (SLTEC) Escherichia coli strains are one of the most important food borne emerging pathogens. One hundred and fifty-seven E. coli strains isolated from 39 children with diarrhea of unknown origin and one hundred and five E. coli strains from 20 healthy children were examined for Shiga-like toxin production in Vero cell line assay. The synthesis of Shiga-like toxin was observed on Vero cell line and confirmed by PCR for one of 262 E. coli strains tested. The shiga-like toxin II-positive E. coli strain was isolated from 2-years old healthy child with no symptoms of gastrointestinal tract infection.

Prevalence of non-O157 Escherichia coli strains among shiga-like toxin-producing (SLTEC) isolates in the region of Lower Silesia, Poland.

During 5 y (1997-2002) in the region of Lower Silesia, Poland, 55 shiga-like toxin-producing strains were isolated from children with diarrhoea, none of whom developed haemolytic uremic syndrome. Shiga-like toxin production was detected on Vero cells and confirmed by the detection of slt1 and slt2 gene sequences. Most of isolates examined possessed slt2 gene alone (41.8%), or in combination with slt1 gene (38.2%). None of the shiga-like toxin-producing strains was of serotype O157:H7. It appears that non-O157 shiga-like toxin-producing E. coil strains are prevalent as the causal agent of severe diarrhoea in the region.

In vitro oxidative activity of cupric complexes of kanamycin A in comparison to in vivo bactericidal efficacy.

The interactions of copper(II) complexes of kanamycin A with oxidation-susceptible biomolecules: 2'-deoxyguanosine, plasmid DNA and yeast tRNA(Phe) were studied in both the presence and absence of hydrogen peroxide. The mixture of complex with H(2)O(2) was found to be an efficient oxidant, converting dG to its 8-oxo derivative, generating strand breaks in plasmid DNA and multiple cleavages in tRNA(Phe). Some of these reactions may play a role in toxic effects of aminoglycoside antibiotics. These complexes were screened for their antibacterial activity. The microbiological studies undertaken to compare the bactericidal action of kanamycin A alone and complexed with copper(II) ions in both neutral and oxidative environment revealed that the enhancement of bactericidal action by Cu(II) was not statistically significant.