Biopolymer encapsulated silver nitrate nanoparticle substrates with surface-enhanced Raman spectroscopy (SERS) for Salmonella detection from chicken rinse.

Salmonella is commonly found on broiler chickens during processing. This study investigates the Salmonella detection method that reduces the necessary time for confirmation, by collecting surface-enhanced Raman spectroscopy (SERS) spectra from bacteria colonies, applied to a substrate of biopolymer encapsulated AgNO3 nanoparticles. Chicken rinses containing Salmonella Typhimurium (ST) were analyzed by SERS and compared to traditional plating and PCR analyses. SERS spectra from confirmed ST and non-Salmonella colonies appear similar in spectra composition, but with different peak intensities. t-Test on the peak intensities showed that ST and non-Salmonella colonies were significantly different (α = 0.0045) at 5 peaks, 692 cm-1, 718 cm-1, 791 cm-1, 859 cm-1, and 1018 cm-1. A support vector machine (SVM) classification algorithm was able to separate ST and non-Salmonella samples with an overall classification accuracy of 96.7 %.

Antibacterial 3D bone scaffolds for tissue engineering application.

Open bone fractures are not only difficult to heal but also are at a high risk of infections. Annual cases of fractures which result from osteoporosis amount to approximately 9 million. Endogenously released nitric oxide (NO) has been shown to play a role in osteogenic differentiation in addition to eradicating infection against a wide variety of pathogens. In the current work, antimicrobial NO releasing 3D bone scaffolds were fabricated using S-nitroso-N-acetyl-penicillamine (SNAP) as the NO donor. During fabrication, nano-hydroxyapatite (nHA) was added to each of the scaffolds in the concentration range of 10-50 wt % in nHA-starch-alginate and nHA-starch-chitosan scaffolds. The mechanical strength of the scaffolds increased proportionally to the concentration of nHA and 50 wt % nHA-starch-alginate possessed the highest load bearing capacity of 203.95 ± 0.3 N. The NO flux of the 50 wt % nHA-starch-alginate scaffolds was found to be 0.50 ± 0.06 × 10-10 mol/min/mg initially which reduced to 0.23 ± 0.02 × 10-10 over a 24 h period under physiological conditions. As a result, a 99.76% ± 0.33% reduction in a gram-positive bacterium, Staphylococcus aureus and a 99.80% ± 0.62% reduction in the adhered viable colonies of gram-negative bacterium, Pseudomonas aeruginosa were observed, which is a significant stride in the field of antibacterial natural polymers. The surface morphology and pore size were observed to be appropriate for the potential bone cell growth. The material showed no toxic response toward mouse fibroblast cells. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1068-1078, 2019.

Improved thermal stability of cellulose nanofibrils using low-concentration alkaline pretreatment.

The thermal stability of cellulose nanofibrils (CNFs) can be improved by converting cellulose crystalline structure to cellulose II using an alkaline treatment method. The conventional method requires around 20wt.% NaOH solutions and causes the cellulose interdigitation and aggregation, making CNFs production difficult. The objective of this study is to develop a new pretreatment method using a low-concentration alkaline solution to produce well-dispersed CNFs with improved thermal stability. CNFs with 90nm diameter were successfully prepared by treating cellulose powder with 2wt.% NaOH solution below 0°C, followed by homogenization through a French pressure cell press. The CNFs had relatively high thermal stability with the mean onset and maximum thermal decomposition temperature of 305°C and 343°C, respectively, compared with the CNFs prepared without the NaOH pretreatment (283°C and 310°C). The increased thermal stability can create new opportunities for the development of CNF-based bio-composites and electronics.

Antimicrobial and Physicochemical Characterization of Biodegradable, Nitric Oxide-Releasing Nanocellulose-Chitosan Packaging Membranes.

Biodegradable composite membranes with antimicrobial properties consisting of nanocellulose fibrils (CNFs), chitosan, and S-nitroso-N-acetyl-d-penicillamine (SNAP) were developed and tested for food packaging applications. As a nitric oxide donor, SNAP was encapsulated into completely dispersed chitosan in 100 mL of 0.1 N acetic acid and was thoroughly mixed with CNFs to produce a composite membrane. The fabricated membranes had a uniform dispersion of chitosan and SNAP within the CNFs, which was confirmed through scanning electron microscopy (SEM) micrographs and a chemiluminescence nitric oxide analyzer. The membranes prepared without SNAP showed lower water vapor permeability than that of the membranes with SNAP. The addition of SNAP resulted in a decrease in Young's modulus for both two- and three-layer membrane configurations. Antimicrobial property evaluation of SNAP-incorporated membranes showed an effective zone of inhibition against bacterial strains of Enterococcus faecalis, Staphylococcus aureus, and Listeria monocytogenes and demonstrated its potential applications for food packaging.

Stable silver/biopolymer hybrid plasmonic nanostructures for high performance surface enhanced Raman scattering (SERS).

Silver/biopolymer nanoparticles were prepared by adding 100 mg silver nitrate to 2% polyvinyl alcohol solution and reduced the silver nitrate using 2% trisodium citrate for high performance Surface Enhanced Raman Scattering (SERS) substrates. Optical properties of nanoparticle were measured using UV/VIS spectroscopy and hyperspectral imaging microscopy. Nanoparticle morphology was analyzed using transmission electron microscopy. Substrate reproducibility and repeatability was checked by measuring SERS signals of trans-1,2-bis(4-pyridyl)ethylene (BPE) and Rhodamine 6G.

Surface enhanced Raman scattering (SERS) with biopolymer encapsulated silver nanosubstrates for rapid detection of foodborne pathogens.

A biopolymer encapsulated with silver nanoparticles was prepared using silver nitrate, polyvinyl alcohol (PVA) solution, and trisodium citrate. It was deposited on a mica sheet to use as SERS substrate. Fresh cultures of Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus and Listeria innocua were washed from chicken rinse and suspended in 10 ml of sterile deionized water. Approximately 5 µl of the bacterial suspensions was placed on the substrate individually and exposed to 785 nm HeNe laser excitation. SERS spectral data were recorded over the Raman shift between 400 and 1800 cm(-1) from 15 different spots on the substrate for each sample; and three replicates were done on each bacteria type. Principal component analysis (PCA) model was developed to classify foodborne bacteria types. PC1 identified 96% of the variation among the given bacteria specimen, and PC2 identified 3%, resulted in a total of 99% classification accuracy. Soft Independent Modeling of Class Analogies (SIMCA) of validation set gave an overall correct classification of 97%. Comparison of the SERS spectra of different types of gram-negative and gram-positive bacteria indicated that all of them have similar cell walls and cell membrane structures. Conversely, major differences were noted around the nucleic acid and amino acid structure information between 1200 cm(-1) and 1700 cm(-1) and at the finger print region between 400 cm(-1) and 700 cm(-1). Silver biopolymer nanoparticle substrate could be a promising SERS tool for pathogen detection. Also this study indicates that SERS technology could be used for reliable and rapid detection and classification of food borne pathogens.

Classification and structural analysis of live and dead Salmonella cells using Fourier transform infrared spectroscopy and principal component analysis.

Fourier transform infrared spectroscopy (FT-IR) was used to detect Salmonella Typhimurium and Salmonella Enteritidis food-borne bacteria and to distinguish between live and dead cells of both serotypes. Bacteria cells were prepared in 10(8) cfu/mL concentration, and 1 mL of each bacterium was loaded individually on the ZnSe attenuated total reflection (ATR) crystal surface (45° ZnSe, 10 bounces, and 48 mm × 5 mm effective area of analysis on the crystal) and scanned for spectral data collection from 4000 to 650 cm(-1) wavenumber. Analysis of spectral signatures of Salmonella isolates was conducted using principal component analysis (PCA). Spectral data were divided into three regions such as 900-1300, 1300-1800, and 3000-2200 cm(-1) based on their spectral signatures. PCA models were developed to differentiate the serotypes and live and dead cells of each serotype. Maximum classification accuracy of 100% was obtained for serotype differentiation as well as for live and dead cells differentiation. Soft independent modeling of class analogy (SIMCA) analysis was carried out on the PCA model and applied to validation sample sets. It gave a predicted classification accuracy of 100% for both the serotypes and its live and dead cells differentiation. The Mahalanobis distance calculated in three different spectral regions showed maximum distance for the 1800-1300 cm(-1) region, followed by the 3000-2200 cm(-1) region, and then by the 1300-900 cm(-1) region. It showed that both of the serotypes have maximum differences in their nucleic acids, DNA/RNA backbone structures, protein, and amide I and amide II bands.

Porous scaffold of gelatin-starch with nanohydroxyapatite composite processed via novel microwave vacuum drying.

Hydroxyapatite (HA) is a fundamental mineral-based biomaterial, used for preparing composites for bone repair and regeneration. Gelatin blended with starch results in scaffold composites with enhanced mechanical properties. A gelatin-starch blend reinforced with HA nanocrystals (nHA) gave biocompatible composites with enhanced mechanical properties. In this study, a porous scaffold of gelatin-starch-nHA composites was fabricated through microwave vacuum drying and crosslinking using trisodium citrate. Three different composite scaffolds were prepared at three different percentages of nHA: 20%, 30% and 40%. The microstructures and compositions of the composites were analyzed. Within the porous structure, the nHA crystals were observed to precipitate. The interaction between the gelatin-starch network film and nHA crystalline material was studied using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction analysis (XRD). XRD reflections showed that there are two different minerals present in the scaffold composite. There were strong reflection peaks close to the 26 degrees and 32 degrees 2theta angles of HA, and close to the 8 degrees and 49 degrees 2theta angles for sodium citrate minerals. The FTIR result suggested that carboxyl groups, C=O and amino groups play crucial roles in HA formation on the surface of a gelatin network.