Selection of G-rich ssDNA aptamers for the detection of enterotoxins of the cholera toxin family.

Cholera and milder diarrheal disease are caused by Vibrio cholerae and enterotoxigenic Escherichia coli and are still a prominent public health concern. Evaluation of suspicious isolates is essential for the rapid containment of acute diarrhea outbreaks or prevention of epidemic cholera. Existing detection techniques require expensive equipment, trained personnel and are time-consuming. Antibody-based methods are also available, but cost and stability issues can limit their applications for point-of-care testing. This study focused on the selection of single stranded DNA aptamers as simpler, more stable and more cost-effective alternatives to antibodies for the co-detection of AB5 toxins secreted by enterobacteria causing acute diarrheal infections. Cholera toxin and Escherichia coli heat-labile enterotoxin, the key toxigenicity biomarkers of these bacteria, were immobilized on magnetic beads and were used in a SELEX-based selection strategy. This led to the enrichment of sequences with a high % GC content and a dominant G-rich motif as revealed by Next Generation Sequencing. Enriched sequences were confirmed to fold into G-quadruplex structures and the binding of one of the most abundant candidates to the two enterotoxins was confirmed. Ongoing work is focused on the development of monitoring tools for potential environmental surveillance of epidemic cholera and milder diarrheal disease.

Facile Preparation and Analytical Utility of ZnO/Date Palm Fiber Nanocomposites in Lead Removal from Environmental Water Samples.

This study reports a facile approach for preparing low-cost, eco-friendly nanocomposites of ZnO nanoparticles (NPs) and date palm tree fiber (DPF) as a biomass sorbent. The hypothesis of this research work is the formation of an outstanding adsorbent based on the date palm fiber and ZnO nanoparticles. ZnO NP/DPF nanocomposites were synthesized by mixing the synthesized ZnO NPs and DPF in different mass ratios and evaluating their efficacy in adsorbing Pb2+ from aqueous solutions. The structure and surface morphology of the developed ZnO NP/DPF nanocomposite were critically characterized by XRD, FESEM, and TEM techniques. Compared to ZnO NPs, the ZnO NP/DPF nanocomposites displayed significantly enhanced Pb2+ uptake. Pb2+ adsorption was confirmed via various isotherm and kinetic models and thermodynamics. The computed Langmuir sorption capacity (qm) was found to be 88.76 mg/g (R2 > 0.998), and the pseudo-second-order R2 > 0.999 model was most appropriate for describing Pb2+ adsorption. Impregnating the biomass with ZnO NPs enhanced the spontaneity of the process, and the value (−56.55 kJ/mol) of ΔH displayed the exothermic characteristics of Pb2+ retention. Only the loaded ZnO NP/DPF achieved the removal of a high percentage (84.92%) of Pb2+ from the environmental water sample (seawater). This finding suggests the use of ZnO NP/DPF nanocomposites for removing heavy metals from environmental water samples to purify the samples.

Silver nanoparticles coated with natural polysaccharides as models to study AgNP aggregation kinetics using UV-Visible spectrophotometry upon discharge in complex environments.

This study provides quantitative information on the aggregation and dissolution behaviour of silver nanoparticles (AgNPs) upon discharge in fresh and sea waters, represented here as NaCl solutions of increasing ionic strength (up to 1M) and natural fjord waters. Natural polysaccharides, sodium alginate (ALG) and gum Arabic (GA), were used as coatings to stabilize the AgNPs and the compounds acted as models to study AgNP aggregation kinetics. The DLVO theory was used to quantitatively describe the interactions between the AgNPs. The stability of AgNPs was established using UV-Visible spectrophotometry, including unique information collected during the first seconds of the aggregaton process. Alginate coating resulted in a moderate stabilization of AgNPs in terms of critical coagulation concentration (~82mM NaCl) and a low dissolution of <10% total Ag in NaCl solutions up to 1M. Gum Arabic coated AgNPs were more strongly stabilized, with ~7-30% size increase up to 77mM NaCl, but only when the silver ion content initially present in solution was low (<10% total Ag). The ALG and GA coated AgNPs showed a strongly enhanced stability in natural fjord waters (ca. 5h required to reduce the area of the surface plasmon resonance band (SPRB) by two fold) compared with NaCl at an equivalent ionic strength (1-2min period for a two fold SPRB reduction). This is ascribed to a stabilizing effect from dissolved organic matter present in natural fjord waters. Interestingly, for AgNP-GA solutions with 40% of total silver present as unreacted silver ions in the NP stock solution, fast aggregation kinetics were observed in NaCl solutions (SPRB area was reduced by ca. 50% within 40-150min), with even more rapid removal in fjord waters, attributed to the high amount of silver-chloride charged species, that interact with the NP coating and/or organic matter and reduce the NPs stabilization.