Mycobacterium paratuberculosis and milk

The possibility that milk from cattle with Johne's disease could be a potential vehicle of transmission of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) to humans has been the focus of a UK government-funded research programme at Queen's University, Belfast since 1993. The main findings of this research programme are reported and practical advice about the most appropriate methods for the isolation/detection of this organism in milk is given. The findings of several milk surveys during which optimised sensitive detection methods were employed (decontamination with 0.75% cetyl pyridinium chloride for 5 h prior to culture and a novel immunomagnetic PCR technique) have revealed that detectable levels of M. paratuberculosis are present in bulked raw cows' milk in the UK at both the farm level and at dairy processing plants prior to pasteurisation. Furthermore, results of three different experimental approaches to assess the effect of pasteurisation time/temperature conditions on the viability of M. paratuberculosis (laboratory pasteurisation studies, a national survey of commercially pasteurised milk, and processing of naturally infected milk through commercial-scale pasteurising plant) provide firm evidence that this organism is capable of surviving commercial milk pasteurisation on occasion. Hence, both raw and pasteurised cows' milk are potential vehicles of transmission of M. paratuberculosis to humans.     

Serological responses against the pathogenic dimorphic fungus Mucor amphibiorum in populations of platypus (Ornithorhynchus anatinus) with and without ulcerative mycotic dermatitis

Mucor amphibiorum, a dimorphic fungus, causes ulcerative dermatitis and systemic infections in the platypus Ornithorhynchus anatinus in some river systems in Tasmania but apparently not in other regions of Australia. As yet there are no suitable tests for population surveys, nor for detection of internal lesions in live animals. Consequently, immunoglobulins were purified from the serum of platypuses and anti-immunoglobulin antisera were prepared in rabbits in order to develop an enzyme-linked immunosorbent assay (ELISA) for anti-M. amphibiorum antibodies. Antigens from plate-grown cultures resulted in greater signal-to-noise ratios in indirect ELISA than those from broth-grown cultures. Platypuses with clinical ulcerative dermatitis had elevated anti-Mucor antibody levels compared to apparently unaffected individuals. Seroconversion was observed in one animal coincident with the development of cutaneous ulcers. The results suggested that platypuses in affected rivers were exposed to M. amphibiorum at a higher frequency than the occurrence of clinical disease. Some platypuses from New South Wales had elevated antibody levels but these increased significantly with age suggesting exposure to cross-reactive antigens, although exposure to M. amphibiorum cannot be excluded. Further studies are warranted to determine factors that result in progression from infection to disease, the occurrence of the fungus in areas where disease has not been observed and the specificity of antigen used in ELISA.     

Outcomes assessment in veterinary medical education

The Virginia-Maryland Regional College of Veterinary Medicine (VMRCVM) agreed to perform outcomes assessment (OA) as part of the accreditation review process for the American Veterinary Medical Association (AVMA). Nine OA instruments were developed and validated by a 20-member accreditation committee. The instruments were also pre-tested by a subset of the target population. The instruments were for alumni one to five years post-graduate, alumni 6-15 years post-graduate, faculty, staff, DVM students, employers of veterinarians, referring veterinarians using the Blacksburg campus, and referring veterinarians using the Leesburg campus. In addition, data were used from OA surveys previously done for the Office of Research and Graduate Studies. Data from the surveys were incorporated into each of the 11 Essentials for Accreditation required by the AVMA. The process of OA provided a comprehensive assessment of the many aspects of the operation of the college. An important follow-up to the OA process is use of data to enhance and/or re-prioritize existing programs.     

Coronavirus-associated epizootic catarrhal enteritis in ferrets

OBJECTIVE: To characterize clinical signs and lesions and identify the etiologic agent associated with epizootic catarrhal enteritis in domestic ferrets. DESIGN: Cross-sectional study. ANIMALS: 119 ferrets with epizootic diarrhea of presumed viral cause and 5 control ferrets. PROCEDURE: Clinical records and biopsy or necropsy specimens of ferrets with presumed epizootic catarrhal enteritis were reviewed. Immunohistochemical staining for coronavirus antigen was performed on paraffin-embedded tissues from approximately 10% of affected ferrets to identify viral antigen and determine its distribution. Transmission electron microscopy was performed on fecal samples and sections of jejunum. Virus isolation studies as well as immunofluorescent tests for other similar viruses were performed. RESULTS: Characteristic microscopic lesions consistent with intestinal coronavirus infection (vacuolar degeneration and necrosis of villus enterocytes; villus atrophy, fusion, and blunting; and lymphocytic enteritis) were consistently detected in affected ferrets. Coronavirus particles were identified in feces and jejunal enterocytes by use of transmission electron microscopy. Immunohistochemical staining of jejunal sections revealed coronavirus antigens. Antigen staining was not detected in healthy ferrets or ferrets with other gastrointestinal tract diseases. Virus isolation was unsuccessful, and other similar viruses were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results strongly implicate a coronavirus as the causative agent of epizootic catarrhal enteritis in ferrets. Diagnosis may be made on the basis of a combination of historical, clinical, and microscopic findings.     

Surveillance for aquabirnavirus in fish hatcheries in Victoria

OBJECTIVE: To determine whether or not aquabirnavirus is present in Victorian fish hatcheries. PROCEDURE: Milt and ovarian fluids were collected from brood stock at 12 hatcheries and cultured in two sensitive cell lines for the presence of viruses. RESULTS: No cytopathic effect was detected indicating the absence of virus. CONCLUSION: There is no evidence of infection with aquabirnavirus or Epizootic Haematopoietic Necrosis virus in Victorian fish hatcheries.     

Application and field validation of a PCR assay for the detection of Mycoplasma hyopneumoniae from swine lung tissue samples.

A PCR assay was validated for the detection of Mycoplasma hyopneumoniae in porcine lung tissue. The detection limit of the assay was 0.18 colony-forming units/g of lung sample spiked with M. hyopneumoniae. In field validation, 426 pigs from 220 cases were examined for M. hyopneumoniae infection by M. hyopneumoniae PCR and a fluorescent antibody (FA) test. In total, 103 pig lungs (24.2%) were positive in the PCR test, and 69 pig lungs (16.2%) were positive in the FA test, among which, 62 pigs were positive for both PCR and FA test. Most of the PCR-positive but FA test-negative cases had lesions compatible with M. hyopneumoniae infection. With Bayesian modeling, the diagnostic sensitivity and specificity of the PCR were determined to be 97.3% and 93.0%, respectively.     

Effects of alternative management practices on the economics,energy and GHG emissions of a wheat–pea cropping system in the Canadian prairies

In recent years alternative farming practices have received considerable attention from Canadian producers as a means to improve their net return from grain and oilseed production. Enhancing the efficiency of nitrogen fertilizer use, including a pulse crop in the rotation, reducing tillage and pesticide use are seen as viable options to reduce reliance on fossil fuel, lower input costs and decrease the risk of soil, air and water degradation. The objective of this study was to determine the effects of 16 alternative management practices for a 2-year spring wheat (Triticum aestivum L.)–field pea (Pisum sativum L.) rotation on economic returns, non-renewable energy use efficiency, and greenhouse gas emissions. The alternative management methods for wheat consisted of a factorial combination of high vs. low soil disturbance one pass seeding, four nitrogen (N) fertilizer rates (20 kg N ha^?1, 40 kg N ha^?1, 60 kg N ha^?1 and 80 kg N ha^?1), and recommended vs. reduced rates of in-crop herbicide application. Alternative management practices for field pea were high vs. low soil disturbance one pass seeding. The resulting 16 cropping systems were evaluated at the whole farm level based on 4 years (two rotation cycles) of data from field experiments conducted on two Orthic Black Chernozem soils (clay loam and loam textures) in Manitoba, Canada. The highest net returns on the clay loam soil were for the high disturbance system with 60 kg N ha^?1 applied to wheat and the recommended rates of in-crop herbicides. The lowest application rate of N, together with low disturbance seeding, provided the highest economic returns on the loam soil. Energy use efficiency was highest for the lowest rate of N application for both tillage systems. The highest rate of N fertilizer and recommended rates of in-crop herbicide produced little additional yield response, lower net returns, and higher GHG emissions. An increase in N fertilizer application from 20 kg ha^?1 to 80 kg ha^?1 increased whole farm energy requirements by about 40%, while reducing herbicide rates had negligible effects on grain yields and total energy input. Overall, as N fertilizer rate increased, the associated GHG emissions were not offset by an increase in carbon retained in the above-ground crop biomass. Moderate to high soil test NO₃-N levels at experimental sites reduced the potential for positive yield responses to N fertilizer in this study, thus minimizing the economic benefits derived from N fertilizer application.     

Modeling First Break Milling of Debranned Wheat Using the Double Normalized Kumaraswamy Breakage Function

Debranning of wheat affects flour quality, initially by altering the breakage of wheat kernels during first break. The double normalized Kumaraswamy breakage function was applied to model the effect of debranning on wheat breakage during first break milling, in which type 1 breakage describes a relatively narrow distribution of midsized particles, whereas type 2 breakage describes a wide size range of predominantly small particles extending to very large particles. Mallacca (hard) and Consort (soft) wheats were debranned and milled at three roll gaps under sharp‐to‐sharp and dull‐to‐dull dispositions. Type 1 breakage increased at longer debranning times, whereas type 2 breakage decreased, for both wheat varieties under both dispositions. Sharp‐to‐sharp milling tended to produce more type 1 breakage than dull‐to‐dull. A mechanism of wheat breakage is proposed to explain the coproduction of very large and small particles via type 2 breakage and, hence, the effect of debranning. The proposed mechanism is that small particles of endosperm arise from scraping of large flat particles of wheat bran under the differential action of the rolls, such that removal of the bran reduces the production of the large bran particles and thus reduces the opportunity for the scraping mechanism that produces the very small particles.     

Direct measurement of the reaction front in chemically amplified photoresists

The continuing drive by the semiconductor industry to fabricate smaller structures using photolithography will soon require dimensional control at length scales comparable to the size of the polymeric molecules in the materials used to pattern them. The current technology, chemically amplified photoresists, uses a complex reaction-diffusion process to delineate patterned areas with high spatial resolution. However, nanometer-level control of this critical process is limited by the lack of direct measurements of the reaction front. We demonstrate the use of x-ray and neutron reflectometry as a general method to measure the spatial evolution of the reaction-diffusion process with nanometer resolution. Measuring compositional profiles, provided by deuterium-labeled reactant groups for neutron scattering contrast, we show that the reaction front within the material is broad rather than sharply defined and the compositional profile is altered during development. Measuring the density profile, we directly correlate the developed film structure with that of the reaction front.