Alena Ševců

Functionalized magnetic nanoparticles and their effect on Escherichia coli and staphylococcus aureus

Magnetite (Fe3O4) nanoparticles were prepared using coprecipitation and subsequently surface-functionalized with 3- aminopropyltriethoxysilane (APTS), polyethylene glycol (PEG), and tetraethoxysilane (TEOS). Nanoparticle morphology was characterized using scanning electron microscopy, while structure and stability were assessed through infrared spectroscopy and zeta potential, respectively. Average size of the nanoparticles analysed by dynamic light scattering was 89 nm, 123 nm, 109 nm, and 130 nm for unmodified magnetite and APTS-, PEG-, and TEOS-modified magnetite nanoparticles, respectively. Biological effect was studied on two bacterial strains: Gram-negative Escherichia coli CCM 3954 and Gram-positive Staphylococcus aureus CCM 3953. Most of modified magnetite nanoparticles had a significant effect on S. aureus and not on E. coli, whereas PEG-magnetite nanoparticles displayed no significant effect on the growth rate of either bacteria.

Polyamide 6/chitosan nanofibers as support for the immobilization of Trametes versicolor laccase for the elimination of endocrine disrupting chemicals.

In recent years, there has been an increase in efforts to improve wastewater treatment as the concentration of dangerous pollutants, such as endocrine disrupting chemicals, in wastewater increases. These compounds, which mimic the effect of hormones, have a negative impact on human health and are not easily removed from water. One way to effectively eliminate these pollutants is to use enzymatically activated materials. In this study, we report on the use of laccase from the white rot fungus Trametes versicolor immobilized onto polyamide 6/chitosan (PA6/CHIT) nanofibers modified using two different spacers (bovine serum albumin and hexamethylenediamine). We then tested the ability of the PA6/CHIT-laccase biocatalysts to eliminate a mixture containing 50μM of two endocrine disrupting chemicals: bisphenol A and 17α-ethinylestradiol. The PA6/CHIT nanofiber matrix used in this study not only proved to be a suitable carrier for immobilized and modified laccase but was also efficient in the removal of a mixture of endocrine disrupting chemicals in three treatment cycles.

Fabrication, characterization, and antibacterial properties of electrospun membrane composed of gum karaya, polyvinyl alcohol, and silver nanoparticles

Gum karaya (GK), a natural hydrocolloid, was mixed with polyvinyl alcohol (PVA) at different weight ratios and electrospun to produce PVA/GK nanofibers. An 80 : 20 PVA/GK ratio produced the most suitable nanofiber for further testing. Silver nanoparticles (Ag-NPs) were synthesised through chemical reduction of AgNO3 (at different concentrations) in the PVA/GK solution, the GK hydroxyl groups being oxidised to carbonyl groups, and Ag+ cations reduced to metallic Ag-NPs. These PVA/GK/Ag solutions were then electrospun to produce nanofiber membranes containing Ag-NPs (Ag-MEMs). Membrane morphology and other characteristics were analysed using scanning electron microscopy coupled with energy dispersive X-ray analysis, transmission electron microscopy, and UV-Vis and ATR-FTIR spectroscopy. The antibacterial activity of the Ag-NP solution and Ag-MEM was then investigated against Gram-negative Escherichia coli and Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus. Our results show that electrospun nanofiber membranes based on natural hydrocolloid, synthetic polymer, and Ag-NPs have many potential uses in medical applications, food packaging, and water treatment.

Dual-modality self-heating and antibacterial polymer-coated nanoparticles for magnetic hyperthermia

Multifunctional nanoparticles for magnetic hyperthermia which simultaneously display antibacterial properties promise to decrease bacterial infections co-localized with cancers. Current methods synthesize such particles by multi-step procedures, and systematic comparisons of antibacterial properties between coatings, as well as measurements of specific absorption rate (SAR) during magnetic hyperthermia are lacking. Here we report the novel simple method for synthesis of magnetic nanoparticles with shells of oleic acid (OA), polyethyleneimine (PEI) and polyethyleneimine-methyl cellulose (PEI-mC). We compare their antibacterial properties against single gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria as well as biofilms. Magnetite nanoparticles (MNPs) with PEI-methyl cellulose were found to be most effective against both S. aureus and E. coli with concentration for 10% growth inhibition (EC10) of <150 mg/l. All the particles have high SAR and are effective for heat-generation in alternating magnetic fields.

Microbial degradation of chloroethenes: a review

Contamination by chloroethenes has a severe negative effect on both the environment and human health. This has prompted intensive remediation activity in recent years, along with research into the efficacy of natural microbial communities for degrading toxic chloroethenes into less harmful compounds. Microbial degradation of chloroethenes can take place either through anaerobic organohalide respiration, where chloroethenes serve as electron acceptors; anaerobic and aerobic metabolic degradation, where chloroethenes are used as electron donors; or anaerobic and aerobic co-metabolic degradation, with chloroethene degradation occurring as a by-product during microbial metabolism of other growth substrates, without energy or carbon benefit. Recent research has focused on optimising these natural processes to serve as effective bioremediation technologies, with particular emphasis on (a) the diversity and role of bacterial groups involved in dechlorination microbial processes, and (b) detection of bacterial enzymes and genes connected with dehalogenation activity. In this review, we summarise the different mechanisms of chloroethene bacterial degradation suitable for bioremediation and provide a list of dechlorinating bacteria. We also provide an up-to-date summary of primers available for detecting functional genes in anaerobic and aerobic bacteria degrading chloroethenes metabolically or co-metabolically.

Zero-valent iron particles for PCB degradation and an evaluation of their effects on bacteria, plants, and soil organisms

Two types of nano-scale zero-valent iron (nZVI-B prepared by borohydride reduction and nZVI-T produced by thermal reduction of iron oxide nanoparticles in H2) and a micro-scale ZVI (mZVI) were compared for PCB degradation efficiency in water and soil. In addition, the ecotoxicity of nZVI-B and nZVI-T particles in treated water and soil was evaluated on bacteria, plants, earthworms, and ostracods. All types of nZVI and mZVI were highly efficient in degradation of PCBs in water, but had little degradation effect on PCBs in soil. Although nZVI-B had a significant negative impact on the organisms tested, treatment with nZVI-T showed no negative effect, probably due to surface passivation through controlled oxidation of the nanoparticles.

Dynamics of organohalide-respiring bacteria and their genes following in-situ chemical oxidation of chlorinated ethenes and biostimulation.

Application of Fentons reagent and enhanced reductive dechlorination are currently the most common remediation strategies resulting in removal of chlorinated ethenes. In this study, the influence of such techniques on organohalide-respiring bacteria was assessed at a site contaminated by chlorinated ethenes using a wide spectrum of molecular genetic markers, including 16S rRNA gene of the organohalide-respiring bacteria Dehaloccocoides spp., Desulfitobacterium and Dehalobacter; reductive dehalogenase genes (vcrA, bvcA) responsible for dechlorination of vinyl chloride and sulphate-reducing and denitrifying bacteria. In-situ application of hydrogen peroxide to induce a Fenton-like reaction caused an instantaneous decline in all markers below detection limit. Two weeks after application, the bvcA gene and Desulfitobacterium relative abundance increased to levels significantly higher than those prior to application. No significant decrease in the concentration of a range of chlorinated ethenes was observed due to the low hydrogen peroxide dose used. A clear increase in marker levels was also observed following in-situ application of sodium lactate, which resulted in a seven-fold increase in Desulfitobacterium and a three-fold increase in Dehaloccocoides spp. after 70 days. An increase in the vcrA gene corresponded with increase in Dehaloccocoides spp. Analysis of selected markers clearly revealed a positive response of organohalide-respiring bacteria to biostimulation and unexpectedly fast recovery after the Fenton-like reaction.

Magnetic Poly(N-isopropylacrylamide) Nanocomposites: Effect of Preparation Method on Antibacterial Properties

The most challenging task in the preparation of magnetic poly(N-isopropylacrylamide) (Fe3O4-PNIPAAm) nanocomposites for bio-applications is to maximise their reactivity and stability. Emulsion polymerisation, in situ precipitation and physical addition were used to produce Fe3O4-PNIPAAm-1, Fe3O4-PNIPAAm-2 and Fe3O4-PNIPAAm-3, respectively. Their properties were characterised using scanning electron microscopy (morphology), zeta-potential (surface charge), thermogravimetric analysis (stability), vibrating sample magnetometry (magnetisation) and dynamic light scattering. Moreover, we investigated the antibacterial effect of each nanocomposite against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. Both Fe3O4-PNIPAAm-1 and Fe3O4-PNIPAAm-2 nanocomposites displayed high thermal stability, zeta potential and magnetisation values, suggesting stable colloidal systems. Overall, the presence of Fe3O4-PNIPAAm nanocomposites, even at lower concentrations, caused significant damage to both E. coli and S. aureus DNA and led to a decrease in cell viability. Fe3O4-PNIPAAm-1 displayed a stronger antimicrobial effect against both bacterial strains than Fe3O4-PNIPAAm-2 and Fe3O4-PNIPAAm-3. Staphylococcus aureus was more sensitive than E. coli to all three magnetic PNIPAAm nanocomposites.

Different effects of nano-scale and micro-scale zero-valent iron particles on planktonic microorganisms from natural reservoir water

While nano-scale and micro-scale zero-valent iron (nZVI and mZVI) particles show high potential for remediation of polluted soil aquifers and elimination of cyanobacterial blooms, this has required their release into the environment. This study compares the impact of 100 mg L−1 of nZVI and mZVI on natural planktonic microorganisms from a reservoir, incubated in 1.5 L batches over 21 days. In addition to counting cyanobacterial and algal cell numbers, bacterial community structure was assessed using Ion Torrent sequencing and the number of cultivable bacteria determined using standard cultivation methods. Surprisingly, while mZVI had no significant effect on algal cell number, cyanobacteria numbers increased slightly after 14 days (P < 0.05). Algae were only marginally affected by nZVI after seven days (P < 0.05), while cyanobacteria numbers remained unaffected after 21 days. Total species richness and less common bacteria increased significantly when treated with mZVI (compared to nZVI). The abundance of Limnohabitans (Betaproteobacteria), Roseiflexus (Chloroflexi), hgcl_clade (Actinobacteria) and Comamonadaceae_unclassified (Betaproteobacteria) increased under nZVI treatment, while mZVI enhanced Opitutae_vadinHA64 (Verrucomicrobia) and the OPB35_soil_group (Verrucomicrobia). Interestingly, the number of cultivable bacteria increased significantly after three days in water with nZVI, and further still after seven days. nZVI shaped bacterial community both directly, through release of Fe(II)/Fe(III), and indirectly, through rapid oxygen consumption and establishment of reductive conditions. The strong physico-chemical changes caused by nZVI proved temporary; hence, it can be assumed that, under natural conditions in resilient reservoirs or lakes, microbial plankton would recover within days or weeks.

A poly(3-hydroxybutyrate)–chitosan polymer conjugate for the synthesis of safer gold nanoparticles and their applications

A facile and eco-friendly approach is developed for the synthesis of a poly(3-hydroxybutyrate)–chitosan (PHB–chit) polymer conjugate, which is an ideal material for the synthesis of safer gold nanoparticles (nAu). The synthesized products are characterized by various electron-based, optical and spectroscopic techniques including the catalytic activity and the stability/toxicity of nAu. In contrast to the conventional synthesis approaches, the use of PHB–chit results in generation of a highly stable and size-controlled nAu material that exhibits important catalytic activity towards the reduction of 4-nitrophenol (4-NP) to 4-aminophenol, and at the same time shows no toxicity against living bacterial systems such as Escherichia coli or Staphylococcus aureus. The synthesis route reported herein would inspire future developments that circumvent toxicity issues of relevance to living systems while performing common catalytic experimentation.