Comparison of Different Leaf Preservation Methods To Obtain High Quality DNA From Enset (Ensete Ventricosum (Welw.) Cheesman), A Native And Orphan Food Security Crop In Ethiopia

Background: Enset (Ensete ventricosum (Welw.) Cheesman) is a staple food for more than 20 million Ethiopians and only cultivated in the native indigenous farming systems of Ethiopia. In contrast to other cultivated species in the Musaceae family, enset has been relatively little studied at the molecular level. Application of advanced molecular genetic techniques requires rapid extraction of DNA of high quality and quantity. Fresh, lyophilized tissues, as well as tissues stored in liquid nitrogen are mainly preferred to avoid DNA degradation, thus most of the DNA extraction protocols recommend these types of tissues as starting material. However, such sample processing techniques are difficult to utilize in many developing countries and at collection sites of many endemic plant species, underutilized or orphan crop species like enset. These situations necessitate the development of alternative protocols for leaf preservation and optimized methods for isolating high-quality DNA from dried or preserved leaf samples. Results: In this study, three different leaf preservation and two DNA extraction methods were compared. Fresh young leaf tissue was preserved using the minor modified saturated NaCl-CTAB solution, silica gel or 96% ethanol at ambient temperature for more than 35 days. Subsequently, DNA was extracted using either the DNeasy Plant Mini Kit or the CTAB method. As compared to silica gel and 96% ethanol, the minor modified saturated NaCl-CTAB solution preserved the quality, quantity, and integrity of enset genomic DNA. This method consistently produced genomic DNA of high-quality and quantity at affordable cost. The DNeasy Plant Mini Kit method was found to be more efficient than the standard CTAB method, being faster and producing genomic DNA of higher quality. Conclusions: Using saturated NaCl-CTAB solution is an accessible, efficient, scalable, and inexpensive way to preserve enset leaves during collection and transportation. The preservation protocol was validated for leaf tissues of all cultivated and wild enset, and Entada landraces. Genomic DNA of high quality and quantity was obtained from preserved enset leaves, which can be used for further downstream applications including PCR and sequencing.

Spheroid Nickel Nanoparticles Synthesized in CTAB Solution Using Gamma Radiation

Pure nickel nanoparticles with some paired grain shaped has been successfully synthesized using gamma radiation technique in aqueous system at ambient temperature without using reducing agent. Cetyl trimethylammonium bromide was used to prevent oxidation during radiolysis process and help to shape the nickel nanoparticles into spheroid. Synthesized nanoparticles were characterized using X-ray diffraction, tunnelling electron microscopy and vibrating sample magnetometer. The particles formed are crystallized with fcc phase without any oxidation state. The particle size ranging from 20 – 50 nm which consists of unique morphology of paired spheroid. Vibrating sample magnetometer analysis shows that sample has ferromagnetic properties with value of magnetic remanence smaller that bulk due to its size.

Production of Poly-γ-Glutamic Acid (γ-PGA) by Clinical Isolates of Staphylococcus Epidermidis

Background and Objective: Poly-γ-glutamic acid (γ-PGA) is a constituent of the Bacillus anthracis capsule and a potential virulence factor of S. epidermidis. In this study, a methodology for the isolation, purification and quantification of γ-PGA in the isolates was adapted. In addition, the fate of the produced γ-PGA and its antiphagocytic activity were investigated. Methods: The capB gene was investigated by the PCR method in 50 isolates of S. epidermidis. A modified methodology was used for the extraction, purification, and quantification of γ-PGA using Cetyltrimethylammonium Bromide (CTAB) solution. The fate of γ-PGA was determined in Tryptic Soy Broth (TSB) medium, as well as the effect of ethanol, NaCl and KCl on the induction of the polymer production. The ability of neutrophils to phagocyte both FITC-labeled latex particles in the presence of free γ-PGA and S. epidermidis with and without anchored γ-PGA was evaluated by cytometry. Results: The production of γ-PGA was detected in 40 isolates; all of them were capB gene carriers. Free γ-PGA was detected and in the strain, the amount of released γ-PGA in the supernatant was 67% greater than the cell anchored γ-PGA. Phagocytosis tests performed with one γ-PGA producer isolate showed a significant reduction in neutrophil internalization. Conclusion: The adapted methodology was able to detect γ-PGA in the isolates studied. In addition to being found attached to the cell wall, it was demonstrated in this study that γ-PGA can also be found in the culture supernatant. Free γ-PGA did not determine a reduction in the internalization of latex by neutrophils, but cells with anchored γ-PGA showed significant protection against phagocytosis.

Fabrication of magnesium oxide nanoparticles by solvent alteration and their bactericidal applications

MgO nanoparticles are synthesized using water, ethanol and aqueous CTAB solution. The nanoparticles synthesized in ethanol exhibited smallest size, maximum reactive oxygen species generation and maximum antibacterial ability, and low haemolysis.

Modified Silica Adsorbent from Volcanic Ash for Cr(VI) Anionic Removal

In the present research, cetyltrymethyl ammonium bromide (CTAB)-modified silica from Kelud’s volcanic ash has been prepared and examined as adsorbent for removal of the hazardous Cr(VI) anion. The research was initiated with purification of SiO2 from the volcanic ash that was carried out by reacting the volcanic ash with NaOH powder at 900 °C for 2 h, followed by dissolving the ash to water at 100 °C, and then was acidified with HCl 1 M to form hydrogel. By calcination of the hydrogel, silica (SiO2) gel was obtained. The next step was modification of the silica with CTAB, that was performed by interacting the CTAB solution with the gel, in which the concentration of the CTAB was varied. Then the CTAB-modified silica samples were characterized by using FTIR, XRD, and SEM machines. The activity of the adsorbent was examined for adsorption of CrO4= in the solution. The results of the research demonstrate that the amorphous silica gel and the amorphous CTAB-modified silica have been obtained. The CTAB-modified silica was found to possess much higher ability in the adsorption of CrO4= anion, that was 48.90 mg/g, compared to that of the unmodified silica gel, as much 5.68 mg/g. These findings strongly prove that the negative surface of the CTAB-modified silica adsorbent has been successfully formed. Furthermore, it is also observed that increasing concentration of CTAB in SiO2-CTA can promote more effective adsorption of the CrO4= from the solution, but the further enlargement of the CTAB concentration leads to the adsorption decreased, and the highest adsorption was shown by CTAB-modified silica prepared with 0.10 mole of CTAB/1 mole SiO2.

Effect of Citrate and Hexadecyltrimethylammonium Bromide on the Formation of Hydroxyapatite Crystal in a Thermal Aqueous Solution

To calrify the combined effect of soft template and biominormolecule on the formation of hydroxyapatite (HAp), the study on the growth of HAp regulated by citrate and(or) hexadecyltrimethylammonium bromide (CTAB) solution was developed in a thermal aqueous. The as-obtained products were characterized by XRD, FT-IR and TEM, altogether with HRTEM and ED. Plate-like HAp crystals in nanosize with relative weak crystallinity were obtained in the presence of citrate. Nano-needle shape HAp crystals were produced in the presence of CTAB. Rod-like HAp crystals in a uniform nanosize were formed under the regulation of CTAB and citrate, which showed the same orientation as that of CTAB only. However, the possible mechanism of the growth of HAp was discussed in detail. CTAB micelles worked as a soft template modified by citrate, which guided to form HAp crystals in different morphology. The study is benefit to control the formation of HAp for its bioapplication.

Soft Template CTAB-Assisted Synthesis of Mesoporous Ni-B Amorphous Alloy Nanoparticles

Ni-B amorphous alloy particle is a newly promising candidate of catalyst for methanol electrooxidation in Direct Methanol Fuel Cell (DMFC). For enhancing catalytic activity and increasing surface area, mesoporous Ni-B amorphous alloy nanoparticles was manufactured by chemical reduction of NiCl2 and KBH4 via surfactant hexadecyltrimethyl ammonium bromide (CTAB) as an soft template. The effects of the CTAB content and the feeding order on the phase evolution, microstructure, and surface area of the resulting products were investigated. Phase and structure of the samples were investigated using XRD, SEM and BET. Amorphous Ni-B nanoparticles with mesoporous structure were obtained in this study. The CTAB content great impacted the mesoporous structure of the resulting products, although the obviously effect on the phase was not found. The surface area, pore volume and pore diameter of samples improved with the increase of CTAB content. Compared to nanoparticles obtained by CTAB solution added to NiCl2 solution, nanoparticles obtained by NiCl2 solution added to CTAB solution exhibited better mesoporous structure.