Ganoderma lucidum-Mediated Green Synthesis of Silver Nanoparticles with Antimicrobial Activity.

"Green chemistry" is a simple and easily reproductible method that provides nanoparticles characterized by better stability and good dispersion in an aqueous solution. Nanoparticles can be synthesized by algae, bacteria, fungi, and plant extracts. Ganoderma lucidum is a commonly used medicinal mushroom with distinctive biological properties, such as antibacterial, antifungal, antioxidant, anti-inflammatory, anticancer, etc. In this study, aqueous mycelial extracts of Ganoderma lucidum were used to reduce AgNO3 to form silver nanoparticles (AgNPs). The biosynthesized nanoparticles were characterized by UV-visible spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) analysis. The maximum UV absorption was obtained at 420 nm, which represents the specific surface plasmon resonance band for biosynthesized silver nanoparticles. SEM images showed particles as predominantly spherical, while FTIR spectroscopic studies illustrated the presence of functional groups that can support the reducing of ion Ag+ to Ag(0). XRD peaks ratified the presence of AgNPs. The antimicrobial effectiveness of synthesized nanoparticles was tested against Gram-positive and Gram-negative bacterial and yeasts strains. The silver nanoparticles were effective against pathogens, inhibiting their proliferation, and thus reducing the risk to the environment and to public health.

Microbial Removal of Heavy Metals from Contaminated Environments Using Metal-Resistant Indigenous Strains.

Contamination of soil with heavy metals has become a matter of global importance due to its impact on agriculture, environmental integrity, and therefore human health and safety. Several microbial strains isolated from soil contaminated by long-term chemical and petrochemical activities were found to manifest various levels of tolerance to Cr, Pb, and Zn, out of which Bacillus marisflavi and Trichoderma longibrachiatum exhibited above-moderate tolerance. The concentrations of target heavy metals before and after bioremediation were determined using electrochemical screen-printed electrodes (SPE) modified with different nanomaterials. The morpho-structural SEM/EDX analyses confirmed the presence of metal ions on the surface of the cell, with metal uptake being mediated by biosorption with hydroxyl, carboxyl, and amino groups as per FTIR observations. T. longibrachiatum was observed to pose a higher bioremediation potential compared to B. marisflavi, removing 87% of Cr and 67% of Zn, respectively. Conversely, B. marisflavi removed 86% of Pb from the solution, compared to 48% by T. longibrachiatum. Therefore, the fungal strain T. longibrachiatum could represent a viable option for Cr and Zn bioremediation strategies, whereas the bacterial strain B. marisflavi may be used in Pb bioremediation applications.

Highlighting Bacteria with Calcifying Abilities Suitable to Improve Mortar Properties.

Biomineralization, the use of microorganisms to produce calcium carbonate, became a green solution for application in construction materials to improve their strength and durability. The calcifying abilities of several bacteria were investigated by culturing on a medium with urea and calcium ions. The characterization of the precipitates from bacterial cultures was performed using X-ray diffraction, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The formation of carbonate crystals was demonstrated by optical and scanning electron microscopy. Water absorption and compressive strength measurements were applied to mortars embedded with sporal suspension. The efficiency of the supplementation of mortar mixtures with bacterial cells was evaluated by properties, namely the compressive strength and the water absorption, which are in a relationship of direct dependence, the increase in compressive strength implying the decrease in water absorption. The results showed that Bacillus subtilis was the best-performing bacterium, its introduction into the mortar producing an increase in compressive strength by 11.81% and 9.50%, and a decrease in water absorption by 11.79% and 10.94%, after 28 and 56 days of curing, respectively, as compared to standards. The exploitation of B. subtilis as a calcifying agent can be an interesting prospect in construction materials.

Fungal Based Biopolymer Composites for Construction Materials.

Environmental contamination, extensive exploitation of fuel sources and accessibility of natural renewable resources represent the premises for the development of composite biomaterials. These materials have controlled properties, being obtained through processes operated in mild conditions with low costs, and contributing to the valorization of byproducts from agriculture and industry fields. A novel board composite including lignocelullosic substrate as wheat straws, fungal mycelium and polypropylene embedded with bacterial spores was developed and investigated in the present study. The bacterial spores embedded in polymer were found to be viable even after heat exposure, helping to increase the compatibility of polymer with hydrophilic microorganisms. Fungal based biopolymer composite was obtained after cultivation of Ganoderma lucidum macromycetes on a mixture including wheat straws and polypropylene embedded with spores from Bacillus amyloliquefaciens. Scanning electron microscopy (SEM) and light microscopy images showed the fungal mycelium covering the substrates with a dense network of filaments. The resulted biomaterial is safe, inert, renewable, natural, biodegradable and it can be molded in the desired shape. The fungal biocomposite presented similar compressive strength and improved thermal insulation capacity compared to polystyrene with high potential to be used as thermal insulation material for applications in construction sector.

Achievements and Prospects in Electrochemical-Based Biosensing Platforms for Aflatoxin M1 Detection in Milk and Dairy Products.

Aflatoxins, which are mainly produced by Aspergillus flavus and parasiticus growing on plants and products stored under inappropriate conditions, represent the most studied group of mycotoxins. Contamination of human and animal milk with aflatoxin M1, the hydroxylated metabolite of aflatoxin B1, is an important health risk factor due to its carcinogenicity and mutagenicity. Due to the low concentration of this aflatoxin in milk and milk products, the analytical methods used for its quantification have to be highly sensitive, specific and simple. This paper presents an overview of the analytical methods, especially of the electrochemical immunosensors and aptasensors, used for determination of aflatoxin M1.

Tetramethyl-6-carboxyrhodamine quenching-based aptasensing platform for aflatoxin B1: Analytical performance comparison of two aptamers.

In this study, a simple TAMRA (tetramethyl-6-carboxyrhodamine) quenching-based aptasensing platform was designed for the detection of aflatoxin B1 (AFB1). Here, we compared the analytical performance of two aptamer sequences: seqA and seqB. The AFB1 detection was based on the interactions of FAM (carboxyfluorescein)-labeled aptamer with TAMRA-labeled DNA complementary strand in the presence and absence of target analyte. Under optimized experimental conditions, TAMRA-labeled strand quenched the fluorescence response of FAM-labeled aptamer due to the noncovalent interaction between the two DNA strands. The binding of AFB1 induced the complex formation and weakened the interaction between FAM-labeled aptamer and TAMRA-labeled complementary strand, resulting in the fluorescence recovery. By using this principle concept, an assay was constructed for the detection of AFB1. The method exhibited good sensitivity, good selectivity with a limit of detection of 0.2 ng ml(-1), and a wide linear range from 0.25 to 32 ng ml(-1). For real sample application, the aptasensors were tested in beer and wine samples, with good recovery rates obtained for AFB1 detection.

Enhanced sensitive love wave surface acoustic wave sensor designed for immunoassay formats.

We report a Love wave surface acoustic wave (LW-SAW) immunosensor designed for the detection of high molecular weight targets in liquid samples, amenable also for low molecular targets in surface competition assays. We implemented a label-free interaction protocol similar to other surface plasmon resonance bioassays having the advantage of requiring reduced time analysis. The fabricated LW-SAW sensor supports the detection of the target in the nanomolar range, and can be ultimately incorporated in portable devices, suitable for point-of-care testing (POCT) applications.

Sensitive detection of endocrine disrupters using ionic liquid--single walled carbon nanotubes modified screen-printed based biosensors.

Simple and low cost biosensor based on screen-printed electrode for sensitive detection of some alkylphenols was developed, by entrapment of HRP in a nanocomposite gel based on single-walled carbon nanotubes (SWCNTs) and 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF(6)]) ionic liquid. Raman and FTIR spectroscopy, CV and EIS studies demonstrate the interaction between SWCNTs and ionic liquid. The nanocomposite gel, SWCNT-[BMIM][PF(6)] provides to the modified sensor a considerable enhanced electrocatalytic activity toward hydrogen peroxide reduction. The HRP based biosensor exhibits high sensitivity and good stability, allowing a detection of the alkylphenols at an applied potential of -0.2V vs. Ag/AgCl, in linear range from 5.5 to 97.7 µM for 4-t-octylphenol and respectively, between 5.5 and 140 µM for 4-n-nonylphenol, with a response time of about 5s. The detection limit was 1.1 µM for 4-t-octylphenol, and respectively 0.4 µM for 4-n-nonylphenol (S/N=3).

Synergistic effect of mediator-carbon nanotube composites for dehydrogenases and peroxidases based biosensors.

Very sensitive, low cost and reliable NADH and H(2)O(2) sensors were realised and used for development of enzyme based biosensors. The active surface of the electrodes was modified with a nanocomposite obtained by modification of SWNT with a proper mediator: Meldola Blue (for NADH) and Prussian Blue (for H(2)O(2)). Low applied potential of -50 mV vs. Ag/AgCl reference electrode proved the synergistic effect of nanocomposite materials towards NADH and H(2)O(2) detection. Biosensors for malic acid and alkylphenols have been developed, using mediator-functionalized-SWNT-based electrodes and two different classes of enzymes: NAD(+)-dependent dehydrogenases and peroxidases. Immobilization of the enzymes was realised using a series of different procedures - adsorption, Nafion membrane, sol-gel and glutaraldehyde, in order to find the best configuration for a good operational stability. A higher sensitivity comparing with other reported biosensors of about 12.41 mA/M.cm(2) was obtained for l-malic acid biosensor with enzyme immobilised in Nafion membrane. Phenol, 4-t-octylphenol and 4-n-nonylphenol were used as standard compounds for HRP based biosensor. Fast biosensor response and comparable detection limit with HPLC methods were achieved.

Strategies to develop malic acid biosensors based on malate quinone oxidoreductase (MQO).

An amperometric biosensor based on malate quinone oxidoreductase (MQO) was developed for monitoring of the malolactic fermentation of wines. Screen-printed electrodes coupled with appropriate mediators were used as transducers for this novel biosensor. MQO was immobilized by physical entrapment in a photo-cross-linkable poly(vinyl alcohol) polymer (PVA-SbQ) on the surface of the working electrode. Several electrochemical mediators were studied in order to lower the applied potential and minimise the matrix effects. Among them, 2,6-dichlorophenol indophenol (DPIP) and phenazine methosulfate (PMS) were chosen for further development. The working conditions (mediator concentration, applied potential and pH) were optimised for both DPIP and PMS. Detection limits for both types of biosensors were of 5 microM malic acid. Sensitivities obtained for the linear part of the calibration curve were 0.85 and 1.7 mA/M for the biosensors based on DPIP and PMS, respectively. Interferences due to non-specific oxidations were shown to be negligible when using PMS as mediator.