Optimizing gDNA purification and qPCR techniques for specific detection and enumeration of probiotic bacteria

Probiotics are live microorganisms, like bacteria and yeast, which are thought to provide health benefits when ingested. Large manufacturers produce blends of probiotics for supplementing the diets of agriculturally important animals. Commercial probiotic products are labeled with the number of cells present at the time of manufacture. Our assignment was to develop an assay which can be used to quantify the number of specific lactic acid bacteria present in certain probiotic products, thus verifying the number reported on the product labels. This quantification process involves isolating genomic DNA (gDNA) from the given samples and then running the DNA through a quantitative polymerase chain reaction (qPCR). We optimized gDNA isolation by 1) treating these gram-positive bacteria with lysozyme and proteinase K, and 2) increasing the length of cell disruption by bead beating (FastPrep). Results showed that gDNA yields were improved by longer FastPrep treatments, but not by enzyme treatments. Quantitative polymerase chain reactions (qPCR) on isolated gDNA allow researchers to make copies of a short target DNA sequence and monitor its amplification in real time through the use of a fluorescent probe. Using this method, we created standard curves for four species of bacteria which allowed us to correlate cell quantity to threshold cycle. These standard curves allow us to quantify the number of each bacteria present in probiotic mixes of unknown composition

AS-777-14 Resolution on Proposal to Establish the Strawberry Sustainability Research and Education Center

That the Academic Senate endorses the establishment of the Strawberry Sustainability Research Education Center

Regulatory Responsibility in the Atomic Energy Program ( A Symposium)

The regulatory actions taken by federal, state and local governments will exert a great impact on the present and future development of the civilian atomic energy program. All are aware of the tremendous potential that atomic energy holds for mankind. All are equally aware, on the negative side, of the potential radiation hazards associated with the use of radiation and radioactive materials. These hazards must be controlled so that the full realization of atomic energy benefits may be enjoyed

Inside NanoSail-D: A Tiny Satellite with Big Ideas

"Small But Mighty" certainly describes the NanoSail-D experiment and mission. Its unique goals and designs were simple, but the implications of this technology are far reaching. From a tiny 3U CubeSat, NanoSail-D deployed a 10 square meter solar sail. This was the first sail vehicle to orbit the earth and was only the second time a sail was unfurled in space. The NanoSail-D team included: two NASA centers, Marshall and Ames, the universities of Alabama in Huntsville and Santa Clara in California, the Air Force Research Laboratory and many contractors including NeXolve, Gray Research and several others. The collaborative nature was imperative to the success of this project. In addition, the Army Space and Missile Defense Command, the Von Braun Center for Science and Innovation and Dynetics Inc. jointly sponsored the NanoSail-D project. This paper presents in-depth insight into the NanoSail-D development. Its design was a combination of left over space hardware coupled with cutting edge technology. Since this NanoSail-D mission was different from the first, several modifications were necessary for the second NanoSail-D unit. Unforeseen problems arose during refurbishment of the second unit and the team had to overcome these obstacles. Simple interfaces, clear responsibilities and division of effort allowed the team members to work independently on the common goal. This endeavor formed working relationships lasting well beyond the end of this mission. NanoSail-D pushed the technology envelop with future applications for all classes of satellites. NanoSail-D is truly a small but mighty satellite, which may cast a very big shadow for years to come

Diversity of Microorganisms Isolated from Amber

Claims that organisms can be cultured from amber, if substantiated, would be significant contributions to our understanding of the evolution, tenacity, and potential spread of life. Three reports on the isolation of organisms from amber have been published. Cano and Borucki recently reported the isolation of Bacillus sphaericus and Lambert et al. have described a new species designated Staphylococcus succinus from 25–40 million year old Dominican amber. These characterized organisms were phylogenetically distant from extant relatives and the Staphylococcus sp. sufficiently far removed from other extant staphylococci to be considered a new species. Here we report the culture of bacteria from Dominican and previously untested 120 million year old Israeli (Lebanese lode) amber. Twenty-seven isolates from the amber matrix have been characterized by fatty-acid profiles (FAME) and/or 16S rRNA sequencing. We also performed a terminal restriction fragment pattern (TRF) analysis of the original amber before prolonged culture by consensus primer amplification of the 16S rRNA followed by restriction enzyme digestion of the amplicons. Sample TRFs were consistent with a sparse bacterial assemblage and included at least five of the isolated organisms. Finally, we microscopically mapped the internal topography of an amber slice

Optimised In 2

Indium sulphide has been extensively investigated as a component for different kind of photovoltaic devices (organic-inorganic hybrid devices, all inorganic, dye sensitized cells). In this paper, we have optimised the growth conditions of indium sulphide thin films by means of a low cost, versatile deposition technique, like spray pyrolysis. The quality of the deposited films has been characterised by micro-Raman, vis-UV spectroscopy, and atomic force microscopy. Substrate deposition temperature and different postdeposition annealing conditions have been investigated in order to obtain information about the quality of the obtained compound (which crystalline or amorphous phases are present) and the morphology of the deposited films. We have shown that the deposition temperature influences strongly the amount of amorphous phase and the roughness of the indium sulphide films. Optimised postdeposition annealing treatments can strongly improve the final amount of the beta phase almost independently from the percentage of the amorphous phase present in the as deposited films

The Impact of Feeding Cannabidiol (CBD) Containing Treats on Canine Response to a Noise-Induced Fear Response Test

Interest is increasing regarding use of Cannabidiol (CBD) in companion animals due to anecdotal evidence of beneficial behavioral and health effects. The purpose of this investigation was to evaluate the influence of CBD on behavioral responses to fear-inducing stimuli in dogs. Sixteen dogs (18.1 ± 0.2 kg) were utilized in a replicated 4 × 4 Latin square design experiment with treatments arranged in a 2 × 2 factorial, consisting of control, 25 mg CBD, trazodone (100 mg for 10–20 kg BW, 200 mg for 20.1–40 kg BW), and the combination of CBD and trazodone. A fireworks model of noise-induced fear was used to assess CBD effectiveness after 7 d of supplementation. Each test lasted a total of 6 min and consisted of a 3 min environmental habituation phase with no noise and a 3 min noise phase with a fireworks track. Plasma was collected 1 h before, immediately after, and 1 h following testing for cortisol analysis. Behaviors in each 3 min block were video recorded, and heart rate (HR) sensors were fitted for collection of HR and HR variability parameters. Research personnel administering treats and analyzing behavioral data were blinded as to the treatments administered. Data were tested for normality using the UNIVARIATE procedure in SAS, then differences examined using the MIXED procedure with fixed effects of treatment, period, time, and treatment x time interaction. Inactivity duration and HR increased during the first minute of the fireworks track compared with 1 min prior (P \u3c 0.001 and P = 0.011, respectively), indicating the fireworks model successfully generated a fear response. Trazodone lowered plasma cortisol (P \u3c 0.001), which was unaffected by CBD (P = 0.104) or the combination with CBD (P = 0.238). Neither CBD nor trazodone affected the duration of inactivity (P = 0.918 and 0.329, respectively). Trazodone increased time spent with tail relaxed (P = 0.001). CBD tended to increase HR (P = 0.093) and decreased the peak of low- and high-frequency bands (LF and HF, P = 0.011 and 0.022, respectively). These results do not support an anxiolytic effect of CBD in dogs given 1.4 mg CBD/kg BW/d