Improved specificity for detection of Mycobacterium bovis in fresh tissues using IS6110 real-time PCR

<p>Abstract</p> <p>Background</p> <p>Culture of <it>M. bovis </it>from diagnostic specimens is the gold standard for bovine tuberculosis diagnostics in the USA. Detection of <it>M. bovis </it>by PCR in tissue homogenates may provide a simple rapid method to complement bacterial culture. A significant impediment to PCR based assays on tissue homogenates is specificity since mycobacteria other than <it>M. bovis </it>may be associated with the tissues.</p> <p>Results</p> <p>Previously published IS<it>6110 </it>based PCR diagnostic assays, along with one developed in house, were tested against environmental mycobacteria commonly isolated from diagnostic tissues submitted to the National Veterinary Services Laboratory. A real-time PCR assay was developed (IS6110_T) that had increased specificity over other IS<it>6110 </it>based assays. Of the 13 non-tuberculous mycobacteria tested with IS6110_T only <it>M. wolinskyi </it>was positive. Thirty <it>M. bovis </it>infected tissue homogenates and 18 control tissues were used to evaluate the potential for the assay as a diagnostic test. In this small sample, IS6110_T detected 20/30 samples from <it>M. bovis </it>infected animals and 0/18 control tissues.</p> <p>Conclusions</p> <p>The IS6110_T assay provides a PCR based assay system that is compatible with current diagnostic protocols for the detection of <it>M. bovis </it>in the USA and compliments current testing strategies.</p

The Thermal Structure of the Circumstellar Disk Surrounding the Classical Be Star gamma Cassiopeia

We have computed radiative equilibrium models for the gas in the circumstellar envelope surrounding the hot, classical Be star $\gamma$Cassiopeia. This calculation is performed using a code that incorporates a number of improvements over previous treatments of the disk's thermal structure by \citet{mil98} and \citet{jon04}; most importantly, heating and cooling rates are computed with atomic models for H, He, CNO, Mg, Si, Ca, & Fe and their relevant ions. Thus, for the first time, the thermal structure of a Be disk is computed for a gas with a solar chemical composition as opposed to assuming a pure hydrogen envelope. We compare the predicted average disk temperature, the total energy loss in H$\alpha$, and the near-IR excess with observations and find that all can be accounted for by a disk that is in vertical hydrostatic equilibrium with a density in the equatorial plane of $\rho(R)\approx 3$ to $5\cdot 10^{-11} (R/R_*)^{-2.5} \rm g cm^{-3}$. We also discuss the changes in the disk's thermal structure that result from the additional heating and cooling processes available to a gas with a solar chemical composition over those available to a pure hydrogen plasma.Comment: 11 pages, 8 figures high resolution figures available at http://inverse.astro.uwo.ca/sig_jon07.htm

Development of a Multidimensional Proteomic Approach to Detect Circulating Immune Complexes in Cattle Experimentally Infected With Mycobacterium bovis

Objective: To evaluate a high-resolution method to identify pathogen-specific biomarkers in serum of calves infected with Mycobacterium bovis.Methods: Serum samples from four calves infected with M. bovis were collected before and after infection at weeks 9, 14, 15, 31, and 36. Immune-complex-associated mycobacterial antigens in the serum were enriched using an immunochromatography method termed, dual path platform (DPP). All regions of antigen capture zones, that consisted of monospecific rabbit polyclonal antibodies raised against M. tuberculosis lysates, on DPP strips were excised and analyzed by multidimensional proteomics. The resulting proteins were then passed through 4 rigorous peptide quality filters-false-hits, decoys, non-M. tuberculosis complex proteins were all removed followed by individual quality check of those remaining. Peptides were then checked on NCBI's BLASTp for M. tuberculosis complex specificity.Results: Proteins in 2 of the animals passed the multipronged-highly stringent peptide quality analysis. Animal#54 had 7 unique M. tuberculosis complex proteins at week 14 post-infection, while animal#56 had 4 at week 36 post-infection along with 1 immunoglobulin.Conclusion:M. tuberculosis complex -specific peptides identified in this study were identified in 2 animals and at 2 separate time points post infection. Further studies with better enrichment protocols and using larger sample sizes and replications are required to develop a TB-specific diagnostic tool for bovine tuberculosis

Detection of a Hot Subdwarf Companion to the Be Star FY Canis Majoris

The rapid rotation of Be stars may be caused in some cases by past mass and angular momentum accretion in an interacting binary in which the mass donor is currently viewed as a small, hot subdwarf stripped of its outer envelope. Here we report on the spectroscopic detection of such a subdwarf in the Be binary system FY Canis Majoris from the analysis of data acquired by the IUE spacecraft and KPNO Coude Feed Telescope over the course of 16 and 21 years, respectively. We present a double-lined spectroscopic orbit for the binary based upon radial velocities from the IUE spectra and use the orbital solutions with a Doppler tomography algorithm to reconstruct the components' UV spectra. The subdwarf is hot (T_eff = 45+/-5 kK) and has a mass of about 1.3 M_sun and a radius of about 0.6 R_sun. It contributes about 4% as much flux as the Be star does in the FUV. We also present observations of the H-alpha and He I 6678 emission features that are formed in the circumstellar disk of the Be star. Orbital flux and velocity variations in the He I 6678 profile indicate that much of the emission forms along the disk rim facing the hot subdwarf where the disk is probably heated by the incident radiation from the subdwarf. A study of the FUV infall shell lines discovered in the 1980s confirms their episodic presence but reveals that they tend to be found around both quadrature phases, unlike the pattern in Algol binaries. Phase-dependent variations in the UV N V doublet suggest the presence of a N-enhanced wind from the subdwarf and a possible shock-interaction region between the stars where the subdwarf's wind collides with the disk of the Be star.Comment: 46 pages, 15 figures, ApJ in pres

A Multiphase Suzaku Study Of X-Rays From Tau Sco

We obtained relatively high signal-to-noise X-ray spectral data of the early massive star tau Sco (B0.2V) with the Suzaku X-ray Imaging Spectrometer (XIS) instrument. This source displays several unusual features that motivated our study: (1) redshifted absorption in UV P Cygni lines to approximately +250 km s(-1) suggestive of infalling gas, (2) unusually hard X-ray emission requiring hot plasma at temperatures in excess of 10 MK whereas most massive stars show relatively soft X-rays at a few MK, and (3) a complex photospheric magnetic field of open and closed field lines. In an attempt to understand the hard component better, X-ray data were obtained at six roughly equally spaced phases within the same epoch of tau Sco\u27s 41 day rotation period. The XIS instrument has three operable detectors: XIS1 is back-illuminated with sensitivity down to 0.2 keV; XIS0 and XIS2 are front-illuminated with sensitivity only down to 0.4 keV and have an overall less effective area than XIS1. The XIS0 and XIS3 detectors show relatively little variability. In contrast, there is a approximate to 4 sigma detection of a approximate to 4% drop in the count rate of the XIS1 detector at one rotational phase. In addition, all three detectors show a approximate to 3% increase in count rate at the same phase. The most optimistic prediction of X-ray variability allows for a 40% change in the count rate, particularly near phases where we have pointings. Observed modulations in the X-ray light curve on the rotation cycle is an order of magnitude smaller than this, which places new stringent constraints on future modeling of this interesting magnetic massive star

On the presence and absence of disks around O-type stars

(abridged) As the favoured progenitors of long-duration gamma-ray bursts, massive stars may represent our best signposts of individual objects in the early Universe, but special conditions seem required to make these bursters, which might originate from the progenitor's rapid rotation and associated asymmetry. To obtain empirical constraints on the interplay between stellar rotation and wind asymmetry, we perform linear Halpha spectropolarimetry on a sample of 18 spectroscopically peculiar massive O stars, including OVz, Of?p, Oe, and Onfp stars, supplemented by an earlier sample of 20 O supergiants. Despite their rapid rotation (with vsin(i) up to 400 km/s) most O-type stars are found to be spherically symmetric, but with notable exceptions amongst specific object classes. We divide the peculiar O stars into four distinct categories: Groups III and IV include the Oe stars and Onfp stars, which are on the high-end tail of the O star rotation distribution and have in the past been claimed to be embedded in disks. Here we report the detection of a classical depolarization ``line effect'' in the Oe star HD 45314, but the overall incidence of line effects amongst Oe stars is significantly lower (1 out of 6) than amongst Be stars. The chance that the Oe and Be datasets are drawn from the same parent population is negligible (with 95% confidence). This implies there is as yet no evidence for a disk hypothesis in Oe stars, providing relevant constraints on the physical mechanism that is responsible for the Be phenomenon. Finally, we find that 3 out of 4 of the group IV Onfp stars show evidence for complex polarization effects, which are likely related to rapid rotation, and we speculate on the evolutionary links to B[e] stars.Comment: Astronomy & Astrophyics (in press) 12 page

Biopsy and tracheobronchial aspirates as additional tools for the diagnosis of bovine tuberculosis in living European bison (bison bonasus)

CITATION: Didkowska, Anna et al. 2020. Biopsy and tracheobronchial aspirates as additional tools for the diagnosis of bovine tuberculosis in living European bison (bison bonasus). Animals, 10(11):2017, doi:10.3390/ani10112017.The original publication is available at: https://www.mdpi.comENGLISH ABSTRACT: The diagnosis of bovine tuberculosis (BTB) in living wildlife remains a complex problem, and one of particular importance in endangered species like European bison (Bison bonasus). To identify infection and avoid the unnecessary culling of such valuable individuals, current best practice requires the collection and culture of material from living animals, as mycobacteria isolation remains the gold standard in BTB diagnosis. However, such isolation is challenging due to the need for the immobilization and collection of appropriate clinical material, and because of the sporadic shedding of mycobacteria. In the present study, we evaluated the potential of sampling for the detection of BTB in a group of seven living European bison suspected of being infected with Mycobacterium caprae. The specimens were collected both as swabs from the nasal and pharyngeal cavities, tracheobronchial aspirates (TBA), ultrasound-guided biopsies from lateral retropharyngeal lymph nodes, and post mortem, from mandibular, retropharyngeal and mediastinal lymph nodes. Clinical samples were tested for mycobacterial species via mycobacteriological culture and PCR. M. caprae was isolated from collected material in two out of four living infected individuals (TBA, biopsy) and mycobacterialDNAwas detected in three out of four (TBA, pharyngeal swab) bison. This is the first report of isolation of M. caprae in living European bison. Our findings demonstrate the value of diagnostic tests based on both molecular testing and culture in European bison and confirm the respiratory shedding of viable M. caprae in this host species.Publisher's versio

Gas Distribution, Kinematics, and Excitation Structure in the Disks around the Classical Be Stars Beta Canis Minoris and Zeta Tauri

Using CHARA and VLTI near-infrared spectro-interferometry with hectometric baseline lengths (up to 330m) and with high spectral resolution (up to 12000), we studied the gas distribution and kinematics around two classical Be stars. The combination of high spatial and spectral resolution achieved allows us to constrain the gas velocity field on scales of a few stellar radii and to obtain, for the first time in optical interferometry, a dynamical mass estimate using the position-velocity analysis technique known from radio astronomy. For our first target star, Beta Canis Minoris, we model the H+K-band continuum and Br-gamma line geometry with a near-critical rotating stellar photosphere and a geometrically thin equatorial disk. Testing different disk rotation laws, we find that the disk is in Keplerian rotation (v(r)=r^(-0.5+/-0.1)) and derive the disk position angle (140+/-1.7 deg), inclination (38.5+/-1 deg), and the mass of the central star (3.5+/-0.2 M_sun). As a second target star, we observed the prototypical Be star Zeta Tauri and spatially resolved the Br{\gamma} emission as well as nine transitions from the hydrogen Pfund series (Pf14-Pf22). Comparing the spatial origin of the different line transitions, we find that the Brackett (Br-gamma), Pfund (Pf14-17), and Balmer (H-alpha) lines originate from different stellocentric radii (R_cont < R_Pf < R_Br-gamma = R_H-alpha), which we can reproduce with an LTE line radiative transfer computation. Discussing different disk-formation scenarios, we conclude that our constraints are inconsistent with wind compression models predicting a strong outflowing velocity component, but support viscous decretion disk models, where the Keplerian-rotating disk is replenished with material from the near-critical rotating star.Comment: 16 pages, 15 figures, accepted by Ap

Current Welfare Problems Facing Horses in Great Britain as Identified by Equine Stakeholders

Despite growing concerns about the welfare of horses in Great Britain (GB) there has been little surveillance of the welfare status of the horse population. Consequently we have limited knowledge of the range of welfare problems experienced by horses in GB and the situations in which poor welfare occurs. Thirty-one in-depth interviews were conducted with a cross -section of equine stakeholders, in order to explore their perceptions of the welfare problems faced by horses in GB. Welfare problems relating to health, management and riding and training were identified, including horses being under or over weight, stabling 24 hours a day and the inappropriate use of training aids. The interviewees also discussed broader contexts in which they perceived that welfare was compromised. The most commonly discussed context was where horses are kept in unsuitable environments, for example environments with poor grazing. The racing industry and travellers horses were identified as areas of the industry where horse welfare was particularly vulnerable to compromise. Lack of knowledge and financial constraints were perceived to be the root cause of poor welfare by many interviewees. The findings give insight into the range of welfare problems that may be faced by horses in GB, the contexts in which these may occur and their possible causes. Many of the problems identified by the interviewees have undergone limited scientific investigation pointing to areas where further research is likely to be necessary for welfare improvement. The large number of issues identified suggests that some form of prioritisation may be necessary to target research and resources effectively

Crop Updates 2005 - Cereals

This session covers thirty six papers from different authors: WHEAT AGRONOMY 1. Optimum sowing time of new wheat varieties in Western Australia, Darshan Sharma, Brenda Shackley, Mohammad Amjad, Christine M. Zaicou-Kunesch and Wal Anderson, Department of Agriculture 2. Wheat varieties updated in ‘Flowering Calculator’: A model predicting flowering time, B. Shackley, D. Tennant, D. Sharma and C.M. Zaicou-Kunesch, Department of Agriculture 3. Plant populations for wheat varieties, Christine M. Zaicou-Kunesch, Wal Anderson, Darshan Sharma, Brenda Shackley and Mohammad Amjad, Department of Agriculture 4. New wheat cultivars response to fertiliser nitrogen in four major agricultural regions of Western Australia, Mohammad Amjad, Wal Anderson, Brenda Shackley, Darshan Sharma and Christine Zaicou-Kunesch, Department of Agriculture 5. Agronomic package for EGA Eagle Rock, Steve Penny, Department of Agriculture 6. Field evaluation of eastern and western wheats in large-scale farmer’s trials, Mohammad Amjad, Ben Curtis and Veronika Reck, Department of Agriculture 7. New wheat varieties for a changing environment, Richard Richards, CSIRO Plant Industry; Canberra 8. Farmers can profitably minimise exposure to frost! Garren Knell, Steve Curtin and David Sermon, ConsultAg 9. National Variety Trials, Alan Bedggood, Australian Crops Accreditation System; Horsham 10. Preharvest-sprouting tolerance of wheat in the field, T.B. Biddulph1, T.L. Setter2, J.A. Plummer1 and D.J. Mares3; 1Plant Biology; FNAS, University of Western Australia; 2Department of Agriculture, 3School of Agriculture and Wine, University of Adelaide 11. Waterlogging induces high concentration of Mn and Al in wheat genotypes in acidic soils, H. Khabaz-Saberi, T. Setter, I. Waters and G. McDonald, Department of Agriculture 12. Agronomic responses of new wheat varieties in the Northern Agricultural Region, Christine M. Zaicou-Kunesch and Wal Anderson, Department of Agriculture 13. Agronomic responses of new wheat varieties in the Central Agricultural Region of WA, Darshan Sharma, Steve Penny and Wal Anderson, Department of Agriculture 14. EGA Eagle Rock tolerance to metribuzin and its mixtures, Harmohinder Dhammu, David Nicholson and Chris Roberts, Department of Agriculture 15. Herbicide tolerance of new bread wheats, Harmohinder Dhammu1 and David Nicholson2, Department of Agriculture NUTRITION 16. The impact of fertiliser placement, timing and rates on nitrogen-use efficiency, Stephen Loss, CSBP Ltd 17. Cereals deficient in potassium are most susceptible to some leaf diseases, Ross Brennan and Kith Jayasena, Department of Agriculture 18. Responses of cereal yields to potassium fertiliser type, placement and timing, Eddy Pol, CSBP Limited 19. Sulphate of Potash, the potash of choice at seeding, Simon Teakle, United Farmers Co-operative 20. Essential disease management for successful barley production, K. Jayasena, R. Loughman, C. Beard, B. Paynter, K. Tanaka, G. Poulish and A. Smith, Department of Agriculture 21. Genotypic differences in potassium efficiency of wheat, Paul Damon and Zed Rengel, Faculty of Natural and Agricultural Sciences, University of Western Australia 22. Genotypic differences in potassium efficiency of barley, Paul Damon and Zed Rengel, Faculty of Natural and Agricultural Sciences, University of Western Australia 23. Investigating timing of nitrogen application in wheat, Darshan Sharma and Lionel Martin, Department of Agriculture, and Muresk Institute of Agriculture, Curtin University of Technology 24. Nutrient timing requirements for increased crop yields in the high rainfall cropping zone, Narelle Hill, Ron McTaggart, Dr Wal Anderson and Ray Tugwell, Department of Agriculture DISEASES 25. Integrate strategies to manage stripe rust risk, Geoff Thomas, Robert Loughman, Ciara Beard, Kith Jayasena and Manisha Shankar, Department of Agriculture 26. Effect of primary inoculum level of stripe rust on variety response in wheat, Manisha Shankar, John Majewski and Robert Loughman, Department of Agriculture 27. Disease resistance update for wheat varieties in WA, M. Shankar, J.M. Majewski, D. Foster, H. Golzar, J. Piotrowski and R. Loughman, Department of Agriculture 28. Big droplets for wheat fungicides, Rob Grima, Agronomist, Elders 29. On farm research to investigate fungicide applications to minimise leaf disease impacts in wheat, Jeff Russell and Angie Roe, Department of Agriculture, and Farm Focus Consultants PESTS 30. Rotations for nematode management, Vivien A. Vanstone, Sean J. Kelly, Helen F. Hunter and Mena C. Gilchrist, Department of Agriculture 31. Investigation into the adaqyacy of sealed farm silos in Western Australia to control phosphine-resistant Rhyzopertha dominica, C.R. Newman, Department of Agriculture 32.Insect contamination of cereal grain at harvest, Svetlana Micic and Phil Michael, Department of Agriculture 33. Phosure – Extending the life of phosphine, Gabrielle Coupland and Ern Kostas, Co-operative Bulk Handling SOIL 34. Optimum combinations of ripping depth and tine spacing for increasing wheat yield, Mohammed Hamza and Wal Anderson, Department of Agriculture 35. Hardpan penetration ability of wheat roots, Tina Botwright Acuña and Len Wade, School of Plant Biology, University of Western Australia MARKETS 36. Latin America: An emerging agricultural powerhouse, Ingrid Richardson, Food and Agribusiness Research, Rabobank; Sydne