Comparative analysis of Bacillus weihenstephanensis KBAB4 spores obtained at different temperatures

The impact of Bacillus weihenstephanensis KBAB4 sporulation temperature history was assessed on spore heat resistance, germination and outgrowth capacity at a temperature range from 7 to 30 °C. Sporulation rate and efficiency decreased at low temperature, as cells sporulated at 12, 20 and 30 °C with approximately 99% efficiency, whereas at 7 °C and 10 °C, a maximum 15% of sporulation was reached. Spores formed at 30 °C showed the highest wet heat resistance at 95 °C, with spores formed at 7 and 10 °C displaying only survival of 15 min exposure at 70 °C, indicating their low level heat resistance. RT-PCR analysis revealed expression of sporulation sigma factor sigG, and germinant receptor operons gerI, gerK, gerL, gerR, gerS, and (plasmid-located) gerS2 to be activated in all sporulation conditions tested. Subsequent germination assays revealed a combination of inosine and L-Alanine to be very efficient, triggering over 99% of the spores to germinate, with spores obtained at 30 °C showing the highest germination rates (99%). Notably, spores obtained at 12, 20 and 30 °C, germinated at all tested temperatures, showing > 70% spore germination even at temperatures as low as 5 °C. Less than 5% of spores obtained at 7 and 10 °C showed a germination response. Furthermore, spores produced at 12, 20 and 30 °C showed similar outgrowth effiency at these temperatures, indicating that low temperature sporulation history does not improve low temperature outgrowth performance. Insights obtained in sporulation and germination behaviour of B. weihenstephanensis KBAB4, in combination with the availability of its genome sequence, may contribute to our understanding of the behaviour of psychrotolerant spoilage and pathogenic Bacill

Design and evaluation of a virtual gearshift application

When a customer buys a new car, he or she wants it to address personal preferences with respect to its driving behavior. By utilizing virtual reality technology, a virtual prototyping environment (VPE) can be created in which the behavior of a vehicle or part of a vehicle can be evaluated and adjusted to match the driver's desires. This paper describes the design and the evaluation of a VPE for manually operated gearboxes. The test group considered the simulated "virtual" gearshift feel to be quite similar to the "real" gearshift feel of a test vehicle. By further developing this VPE, it should become possible to define gearshift feel by customer assessment through haptic simulation, after which the physical gearbox is designed in such a way that it matches the preferred shifting behavior

H-alpha features with hot onsets. II. A contrail fibril

The solar chromosphere observed in H-alpha consists mostly of narrow fibrils. The longest typically originate in network or plage and arch far over adjacent internetwork. We use data from multiple telescopes to analyze one well-observed example in a quiet area. It resulted from the earlier passage of an accelerating disturbance in which the gas was heated to high temperature as in the spicule-II phenomenon. After this passage a dark H-Halpha fibril appeared as a contrail. We use Saha-Boltzmann extinction estimation to gauge the onset and subsequent visibilities in various diagnostics and conclude that such H-alpha fibrils can indeed be contrail phenomena, not indicative of the thermodynamic and magnetic environment when they are observed but of more dynamic happenings before. They do not connect across internetwork cells but represent launch tracks of heating events and chart magnetic field during launch, not at present.Comment: Accepted for Astronomy & Astrophysic

Germinant receptor diversity and germination responses of four strains of the Bacillus cereus group

Four strains of the Bacillus cereus group were compared for their germinant receptor composition and spore germination capacity. Phylogenetic analysis of the germinant receptor encoding operons of the enterotoxic strains B. cereus ATCC 14579 and ATCC 10987, the emetic strain AH187, and the psychrotolerant strain Bacillus weihenstephanensis KBAB4, indicated a core group of five germinant receptor operons to be present in the four strains, with each strain containing one to three additional receptors. Using quantitative PCR, induction of expression during sporulation was confirmed for all identified germinant receptor operons in these strains. Despite the large overlap in receptors, diversity in amino acid-induced germination capacity was observed, with six out of 20 amino acids, serving as germinants for spores of all four strains. Each strain showed unique features: efficient germination of strain KBAB4 spores required non-inducing amounts of inosine as the co-germinant, strain ATCC 10987 spores germinated only efficiently after heat activation. Furthermore, strain ATCC 14579 and AH187 spores germinated without heat activation or inosine, with strain ATCC 14579 spores being triggered by all amino acids except phenylalanine and strain AH187 spores being specifically triggered efficiently only by phenylalanine. Analysis of all germination data did not reveal strict linkages between specific germinants and germinant receptors. Finally, the diversity in nutrient-induced germination capacity was also reflected in the diverse germination responses of heat-activated spores of the four B. cereus strains in food matrices, such as milk, rice water and meat bouillon, indicating that amino acid composition and/or availability of inosine are important germination determinants in foods. Keywords: Ger operon; Food preservation; B. weihenstephanensis; Sporulatio

Habitat Characteristics and Occupancy Rates of Lewis\u27s Woodpecker in Aspen

Lewis‘ woodpeckers (Melanerpes lewis) are generally associated with open ponderosa pine (Pinus ponderosa), open riparian, and burned pine habitats in the West; however, this species has recently been found to nest in aspen (Populus tremuloides) stands in Utah. This study describes the habitat characteristics of Lewis‘ woodpecker nest sites in aspen and investigates how well aspen stand characteristics predict Lewis‘ woodpecker occupancy. I surveyed for Lewis‘ woodpeckers at previously occupied nesting locations in aspen and took habitat measurements at nest sites. In addition, nest-centered Forest Inventory and Analysis (FIA)-type plots provided stand-level habitat characteristics. I used logistic regression to determine which stand-level habitat variables were associated with nest locations; significant variables were then used to select FIA plots in Utah that contained predicted suitable nesting habitat. Criteria used to select FIA plots were aspen type stands, percent canopy cover less than 46%, and average tree diameter at breast height greater than 27.9 cm (11 inches). I then conducted occupancy surveys at FIA plots predicted to contain “suitable” and “non-suitable” Lewis’ woodpecker habitat to field validate the predictive model. No predicted non-suitable plots (n=26) were occupied and only one predicted suitable plot (n=49) was occupied. My results indicated that Lewis’ woodpeckers are rare throughout Utah in aspen stands even though there seems to be abundant nesting habitat available. My results also indicated that variables measured by FIA do not, in isolation, provide sufficient capability to predict Lewis’ woodpecker nesting habitat or actual use, and that more data are needed to accurately predict Lewis’ woodpecker nesting habitat, such as distance to, age, and severity of fires