Detection of pathogenic bacteria in large volume food samples using an enzyme-linked immunoelectrochemical biosensor
FOOD CONTROL
Authors: Capobianco, Joseph A.; Armstrong, Cheryl M.; Lee, Joe; Gehring, Andrew G.
Abstract
Increased speed and sensitivity of testing are always desired for the detection of pathogens in foods. Presented a test sample with low microbial analyte concentration, it is an advantage to analyze as much volume as possible of the sample to attain the best limit of detection (LOD). Therefore, a rapid screening method using a novel flowthrough immunoelectrochemical biosensor was developed for the detection of pathogenic bacteria (E. coli O157: H7 and Salmonella) in food (ground beef). As the working electrode employed was comprised of a porous, antibody-coated graphite felt electrode that served as both a biorecognition-element coated solid support for capture of targeted pathogens as well as a signal transducer, high volumes of aqueous sample could be rapidly exposed to the solid support via gravity flow. Flow rates as high as 16.7 mL/min and 12.3 mL/min could be achieved for bacterial samples in buffer and 1:4 ground meat (beef) homogenate, respectively, with no significant effect on LOD. Fastest flow rates for beef homogenate, without clogging of the porous electrode as well as reduction in apparent electrochemical interference, was realized with a tandem combination of sample pretreatment strategies that included filtration with glass wool and graphite felt as well as continuous flow centrifugation. The LOD for 10,000 E. coli O157 cells in 5, 60, and 1000 mL of buffer was 2000, 170, and 10 cells/mL, respectively in a total assay time of 3 h whereas the LOD for E. coli O157 was 400 cells/mL in 1:4 beef homogenate.
Bactericidal and In-Vitro Cytotoxic Efficacy of Silver Nanoparticles (Ag-NPs) Fabricated by Endophytic Actinomycetes and Their Use as Coating for the Textile Fabrics
NANOMATERIALS
Authors: Salem, Salem S.; EL-Belely, Ehab F.; Niedbala, Gniewko; Alnoman, Maryam M.; Hassan, Saad El-Din; Eid, Ahmed Mohamed; Shaheen, Tharwat I.; Elkelish, Amr; Fouda, Amr
Abstract
An endophytic strain of Streptomyces antimycoticus L-1 was isolated from healthy medicinal plant leaves of Mentha longifolia L. and used for the green synthesis of silver nanoparticles (Ag-NPs), through the use of secreted enzymes and proteins. UV-vis spectroscopy, Fourier-transform infrared (FT-IR), transmission electron microscopy (TEM), X-ray diffraction (XRD), and dynamic light scattering (DLS) analyses of the Ag-NPs were carried out. The XRD, TEM, and FT-IR analysis results demonstrated the successful biosynthesis of crystalline, spherical Ag-NPs with a particle size of 13-40 nm. Further, the stability of the Ag-NPs was assessed by detecting the surface Plasmon resonance (SPR) at 415 nm for one month or by measuring the NPs surface charge (-19.2 mV) by zeta potential analysis (zeta). The green-synthesized Ag-NPs exhibited broad-spectrum antibacterial activity at different concentrations (6.25-100 ppm) against the pathogens Staphylococcus aureus, Bacillus subtilis Pseudomonas aeruginosa, Escherichia coli, and Salmonella typhimurium with a clear inhibition zone ranging from (9.5 +/- 0.4) nm to (21.7 +/- 1.0) mm. Furthermore, the green-synthesized Ag-NPs displayed high efficacy against the Caco-2 cancerous cell line (the half maximal inhibitory concentration (IC50) = 5.7 +/- 0.2 ppm). With respect to antibacterial and in-vitro cytotoxicity analyses, the Ag-NPs concentration of 100 ppm was selected as a safe dose for loading onto cotton fabrics. The scanning electron microscopy connected with energy-dispersive X-ray spectroscopy (SEM-EDX) for the nano-finished fabrics showed the distribution of Ag-NPs as 2% of the total fabric elements. Moreover, the nano-finished fabrics exhibited more activity against pathogenic Gram-positive and Gram-negative bacteria, even after 10 washing cycles, indicating the stability of the treated fabrics.