Development of super nanoantimicrobials combining AgCl, tetracycline and benzalkonium chloride.

In this work, we demonstrate that a simple argentometric titration is a scalable, fast, green and robust approach for producing AgCl/antibiotic hybrid antimicrobial materials. We titrated AgNO3 into tetracycline hydrochloride (TCH) aqueous solution, thus forming AgCl/TCH in a one-step procedure. Furthermore, we investigated the one-pot synthesis of triply synergistic super-nanoantimicrobials, combining an inorganic source of Ag+ ions (AgCl), a disinfecting agent (benzyl-dimethyl-hexadecyl-ammonium chloride, BAC) and a molecular antibiotic (tetracycline hydrochloride, TCH). Conventional antimicrobial tests, industrial biofilm detection protocols, and in situ IR-ATR microbial biofilm monitoring, have been adapted to understand the performance of the synthesized super-nanoantimicrobial. The resulting hybrid AgCl/BAC/TCH nanoantimicrobials are found to be synergistically active in eradicating Salmonella enterica and Lentilactobacillus parabuchneri bacteria and biofilms. This study paves the way for the development of a new class of super-efficient nanoantimicrobials that combine relatively low amounts of multiple active species into a single (nano)formulation, thus preventing the development of antimicrobial resistance towards a single active principle.

Using staining as reference for spectral imaging: Its application for the development of an analytical method to predict the presence of bacterial biofilms.

At present, although spectral imaging is known to have a great potential to provide a massive amount of valuable information, the lack of reference methods remains as one of the bottlenecks to access the full capacity of this technique. This work aims to present a staining-based reference method with digital image treatment for spectral imaging, in order to propose a fast, efficient, contactless and non-invasive analytical method to predict the presence of biofilms. Spectral images of Pseudomonasaeruginosa biofilms formed on high density polyethylene coupons were acquired in visible and near infrared (vis-NIR) range between 400 and 1000 nm. Crystal violet staining served as a biofilm indicator, allowing the bacterial cells and the extracellular matrix to be marked on the coupon. Treated digital images of the stained biofilms were used as a reference. The size and pixels of the hyperspectral and digital images were scaled and matched to each other. Intensity color thresholds were used to differentiate the pixels associate to areas containing biofilms from those ones placed in biofilm-free areas. The model facultative Gram-negative bacterium, P. aeruginosa, which can form highly irregularly shaped and heterogeneous biofilm structures, was used to enhance the strength of the method, due to its inherent difficulties. The results showed that the areas with high and low intensities were modeled with good performance, but the moderate intensity areas (with potentially weak or nascent biofilms) were quite challenging. Image processing and artificial neural networks (ANN) methods were performed to overcome the issues resulted from biofilm heterogeneity, as well as to train the spectral data for biofilm predictions.

Highly Stable Core-Shell Nanocolloids: Synergy between Nano-Silver and Natural Polymers to Prevent Biofilm Formation.

Active investment in research time in the development and study of novel unconventional antimicrobials is trending for several reasons. First, it is one of the ways which might help to fight antimicrobial resistance and bacterial contamination due to uncontrolled biofilm growth. Second, minimizing harmful environmental outcomes due to the overuse of toxic chemicals is one of the highest priorities nowadays. We propose the application of two common natural compounds, chitosan and tannic acid, for the creation of a highly crosslinked polymer blend with not only intrinsic antimicrobial properties but also reducing and stabilizing powers. Thus, the fast and green synthesis of fine spherically shaped Ag nanoparticles and further study of the composition and properties of the colloids took place. A positively charged core-shell nanocomposition, with an average size in terms of the metal core of 17 ± 4 nm, was developed. Nanoantimicrobials were characterized by several spectroscopic (UV-vis and FTIR) and microscopic (transmission and scanning electron microscopies) techniques. The use of AgNPs as a core and an organic polymer blend as a shell potentially enable a synergistic long-lasting antipathogen effect. The antibiofilm potential was studied against the food-borne pathogens Salmonella enterica and Listeria monocytogenes. The antibiofilm protocol efficiency was evaluated by performing crystal violet assay and optical density measurements, direct visualization by confocal laser scanning microscopy and morphological studies by SEM. It was found that the complex nanocomposite has the ability to prevent the growth of biofilm. Further investigation for the potential application of this stable composition in food packaging will be carried out.