Structure and Temperature Regulated Expression of a Cysteine Proteinase Gene in Pachysandra terminalis Sieb. & Zucc.

A cysteine proteinase gene (DQ403257) with an open reading frame of 1125 base pairs was isolated from Pachysdandra terminalis. The primary translated peptide has a predicted length of 374 amino acids, pI (isoelectric point) of 5.70, and molecular mass of 40.9 kDa. The Peptidase_C1 domain is between residue 141 and 367. The proteinase has a conserved motif Gly-Xaa-Thy-Xaa-Phe-Xaa-Asn in the pro region. Sequence comparison shows that the deduced peptide shares 82% identity with the cysteine proteinase RD19a precursor (RD19) (accession P43296) from Arabidopsis thaliana (L.) Heynh. Real-time quantitative reverse-transcriptase–polymerase chain reaction revealed that the gene is induced by treatments of 1 to 7 days of darkness, 2 hours and 3 to 7 days at 5 °C, and 3 days at 38 °C

Magnetic Nanoparticles Enhanced Surface Plasmon Resonance Biosensor for Rapid Detection of Salmonella Typhimurium in Romaine Lettuce

Salmonella is one of the major foodborne pathogens responsible for many cases of illnesses, hospitalizations and deaths worldwide. Although different methods are available to timely detect Salmonella in foods, surface plasmon resonance (SPR) has the benefit of real-time detection with a high sensitivity and specificity. The purpose of this study was to develop an SPR method in conjunction with magnetic nanoparticles (MNPs) for the rapid detection of Salmonella Typhimurium. The assay utilizes a pair of well-characterized, flagellin-specific monoclonal antibodies; one is immobilized on the sensor surface and the other is coupled to the MNPs. Samples of romaine lettuce contaminated with Salmonella Typhimurium were washed with deionized water, and bacterial cells were captured on a filter membrane by vacuum filtration. SPR assays were compared in three different formats—direct assay, sequential two-step sandwich assay, and preincubation one-step sandwich assay. The interaction of flagellin and MNPs with the antibody-immobilized sensor surface were analyzed. SPR signals from a sequential two-step sandwich assay and preincubation one-step sandwich assay were 7.5 times and 14.0 times higher than the direct assay. The detection limits of the assay were 4.7 log cfu/mL in the buffer and 5.2 log cfu/g in romaine lettuce samples

Kinetic Analysis and Epitope Mapping of Monoclonal Antibodies to Salmonella Typhimurium Flagellin Using a Surface Plasmon Resonance Biosensor

Salmonella Typhimurium is one of the leading causes of foodborne diseases worldwide. Biosensors and immunoassays utilizing monoclonal antibodies are widely used for the detection and subtyping of S. Typhimurium. However, due to insufficient information on the nature of binding with S. Typhimurium flagellin, the selection of appropriate antibodies for assay development is a cumbersome task. Hence, we aimed to compare the binding kinetics of a panel of monoclonal antibodies and their relative binding sites to flagellin antigen using a surface plasmon resonance biosensor. Initially, the flagellin was captured on the sensor surface through an immobilized anti-flagellin antibody. The interactions of different concentrations of monoclonal antibodies to flagellin were determined, and binding curves were fitted using 1:1 bio-interaction model to calculate the kinetic parameters. For epitope mapping, pairwise comparisons were completed to determine the binding inhibition of each paired combination of monoclonal antibodies. It was found that these monoclonal antibodies differed significantly (p \u3c 0.05) in association rate, dissociation rate, and equilibrium dissociation constants. Of the five monoclonal antibodies, only two interfered with the binding of each other. Four distinct epitopes located within a 23 kDa domain of flagellin were identified. Findings from this study provide crucial information needed for the further development and optimization of biosensors and other immunoassays for the detection and subtyping of Salmonella

Prevalence and Antimicrobial Resistance of Pathogenic Bacteria in Chicken and Guinea Fowl

This study was conducted to compare the presence and antimicrobial susceptibility of Campylobacter, Salmonella spp., and other enteric bacteria between chickens and guinea fowls. Birds were reared on enclosed concrete floor housing covered with pine wood shavings litter material. Chicken (n = 40) and guinea fowl (n = 40) carcasses, drinking water (10 mL; n = 40), and litter (10 g; n = 40) were aseptically collected randomly from a poultry farm and analyzed within 1 h of collection. Individual pens served as experimental units and were replicated twice. Campylobacter spp., Salmonella spp., and other enterobactericeae were isolated and identified using standard selective media and biochemical tests. Isolates were tested for sensitivity to tetracycline, ampicillin, streptomycin, kanamycin, nalidixic acid, gentamicin, erythromycin, ciprofloxacin, cefoxitin, and colistin using the Kirby-Bauer disk diffusion test. Campylobacter spp. and Salmonella spp. were isolated from 28 and 35% of whole carcass rinses of chickens and from 18 and 23% of whole carcass rinses of guinea fowl, respectively. Although only Salmonella spp. were recovered from drinking water, both Salmonella and Campylobacter spp. were recovered from litter material. Campylobacter upsaliensis was recovered only in the guinea fowl, whereas Klebsiella oxytoca and Enterobacter sakazakii were recovered only in chickens. Although no antibiotic resistance was determined in Campylobacter upsaliensis, most Campylobacter, Salmonella, and Escherichia coli isolates from both chickens and guinea fowl were resistant to antibiotics such as ampicillin, kanamycin, erythromycin, and nalidixic acid

Development of a Game-Based e-Learning System with Augmented Reality for Improving Students’ Learning Performance

Currently, the school children usually spend a lot of time on the games in their recreational activities and some of them are even addicted to the games. Compared with other extracurricular activities, the e-Learning system reflects the fact that school children are very interested in the games. As a result, educators have lately craved to develop effective teaching activities that allow the school children to learn some subjects and to play the games simultaneously.  Therefore, this study is based on an e-Learning system which combines the serious game by Unity3D Game Engine with augmented reality (AR). Students are able to acquire their knowledge and to foster logical skills via this game-based e-Learning system.  According to its efficacy and utilities, this study has assessed and compared the game-based e-Learning system with the traditional learning and other e-Learning systems. The experimental results have indicated that the proposed game-based e-Learning system can outperform other existing systems

Evaluating the UV-C sensitivity of Coxiella burnetii in skim milk using a bench-scale collimated beam system and comparative thermal sensitivity study by high-temperature short-time pasteurization

Introduction:Coxiella burnetii is a zoonotic Gram-negative obligate intracellular bacterial pathogen and the causative agent of query (Q) fever in humans. Contamination of milk by C. burnetii, as a consequence of livestock infection, is a significant public health concern. Effective methods to inactivate C. burnetii in milk are a critical aspect of food safety. Implementation of non-thermal UV-C processing technologies in the dairy industry can effectively preserve the sensory and nutritional quality of raw milk products while ensuring their safety, making them a viable alternative to traditional high-temperature short-time (HTST) pasteurization methods.Methods: Optical light attenuation factors, such as the absorption, scattering, and reflection by skim milk (SM) were evaluated using a spectrophotometer. SM inoculated with an avirulent strain of C. burnetii was irradiated using a collimated beam device equipped with a low-pressure UV-C 254 nm lamp at doses from 0 to 12 mJ/cm2. Optical properties were considered for the evaluation of the delivered UV-C dose. The pasteurization treatment was conducted using a lab scale HTST pasteurizer (72°C/15 s). The verification studies were conducted using Escherichia coli ATCC 25922 inoculated in a phosphate buffer (transparent fluid) and humic acid (opaque fluid). Salmonella enterica serovar Muenchen ATCC BAA 1674 inoculated in SM was tested for its suitability as a surrogate for C. burnetii, a bacterium that requires specialized equipment and expertise for experimentation.Results and Discussion: Absorption, reduced scattering coefficient, and the reflectance of SM at 254 nm were measured as 19 ± 0.3/cm, 26 ± 0.5/cm, and 10.6%, respectively. The UV-C results showed a log-linear inactivation of C. burnetii in SM with the UV-C sensitivity (D10) value of 4.1 ± 0.04 mJ/cm2. The results of HTST pasteurization revealed that C. burnetii was heat-sensitive with a D value of 1.75 min. Salmonella Muenchen showed similar UV inactivation kinetics and is, thereby, suggested as a suitable surrogate to C. burnetii for the pilot-scale UV-C processing studies of SM

Prime Focus Spectrograph (PFS) for the Subaru Telescope: Overview, recent progress, and future perspectives

PFS (Prime Focus Spectrograph), a next generation facility instrument on the 8.2-meter Subaru Telescope, is a very wide-field, massively multiplexed, optical and near-infrared spectrograph. Exploiting the Subaru prime focus, 2394 reconfigurable fibers will be distributed over the 1.3 deg field of view. The spectrograph has been designed with 3 arms of blue, red, and near-infrared cameras to simultaneously observe spectra from 380nm to 1260nm in one exposure at a resolution of ~1.6-2.7A. An international collaboration is developing this instrument under the initiative of Kavli IPMU. The project is now going into the construction phase aiming at undertaking system integration in 2017-2018 and subsequently carrying out engineering operations in 2018-2019. This article gives an overview of the instrument, current project status and future paths forward.Comment: 17 pages, 10 figures. Proceeding of SPIE Astronomical Telescopes and Instrumentation 201

The Bacteria Content of Bagged, Pre-Washed Greens as Related to the Best if Used by Date

The sale of ready-to-eat salads has increased over the past years, yet little is known about consumer usage and the related safety of these products. This study evaluated the changes of microbiological quality of pre-washed spinach and mixed leafy vegetables during refrigeration storage. Microbial loads were determined by aerobic plate count (APC) and Enterobacteriaceae count (EC). The microbiological quality of pre-washed greens varied widely and deteriorated rapidly in a refrigerator. At “best if used by” date, twenty percent of samples had APC of more than 7.0 Log CFU/g and all samples had EC of more than 5.0 Log CFU/g. It is recommended that consumers purchase and eat pre-washed greens in their entirety as far in advance of the “best if used by” date as possible

Microbiological Quality of Packaged Lunchmeat as Related to the Sell-by-date

Consumers are often confused by the product dating systems used by the food manufacturers. However, they have reported that they consider these dates when purchasing lunchmeats and other ready-to-eat foods. A study was conducted to evaluate changes of microbiological quality of packaged lunchmeat during refrigerated storage as related to the sell-by-date (SBD). Thirty packages of lunchmeat with the same lot number were tested over an extended period. The microbiological quality was satisfactory at the time of purchase. It deteriorated steadily during refrigerated storage regardless of whether the packages were opened or not, and was unsatisfactory at SBD. Food manufacturers should strive to meet the microbiological quality standards and consider the usefulness of the information to consumers when setting a product date