Tuberculin Skin Testing in White-Tailed Deer (<i>Odocoileus virginianusi>)

The comparative cervical skin test for antemortem diagnosis of tuberculosis was done 169 times on 116 different white-tailed deer of known Mycobacterium bovis infection status. The sensitivity and specificity were 97 and 81%, respectively. The magnitude of change in skin thickness at test sites was not significantly influenced by dosage of inoculum, dissemination of the disease process, or repeated skin testing. However, the magnitude of change in skin thickness was significantly greater in deer infected for less than 109 days than in deer infected for more than 109 days. As used in the present study, the comparative cervical skin test is a sensitive method of antemortem diagnosis of M. bovis infection in white-tailed deer.

Evaluation of Blood Assays for Detection of <i>Mycobacterium bovisi> in White-Tailed Deer (<i>Odocoileus virginianusi>) in Michigan

Surveillance and control activities related to bovine tuberculosis (TB) in free-ranging, Michigan white-tailed deer (Odocoileus virginianus) have been underway for over a decade, with significant progress. However, foci of higher TB prevalence on private lands and limited agency ability to eliminate them using broad control strategies have led to development and trial of new control strategies, such as live trapping, testing, and culling or release. Such strategies require a prompt, accurate live animal test, which has thus far been lacking. We report here the ability of seven candidate blood assays to determine the TB infection status of Michigan deer. Our aims were twofold: to characterize the accuracy of the tests using field-collected samples and to evaluate the feasibility of the tests for use in a test-and-cull strategy. Samples were collected from 760 deer obtained via five different surveys conducted between 2004 and 2007. Blood samples were subjected to one or more of the candidate blood assays and evaluated against the results of mycobacterial culture of the cranial lymph nodes. Sensitivities of the tests ranged from 46% to 68%, whereas specificities and negative predictive values were all .92%. Positive predictive values were highly variable. An exploratory analysis of associations among several host and sampling-related factors and the agreement between blood assay and culture results suggested these assays were minimally affected. This study demonstrated the capabilities and limitations of several available blood tests for Mycobacterium bovis on specimens obtained through a variety of field surveillance methods. Although these blood assays cannot replace mass culling, information on their performance may prove useful as wildlife disease managers develop innovative methods of detecting infected animals where mass culling is publicly unacceptable and cannot be used as a control strategy.

<i>Mycobacterium bovisi>–Infected White-Tailed Deer (<i>Odocoileus Virginianusi>): Detection of Immunoglobulin Specific to Crude Mycobacterial Antigens by ELISA

White-tailed deer (Odocoileus virginianus) have recently emerged as a source of Mycobacterium bovis infection for cattle within North America. The objective of this study was to evaluate the antibody response of M. bovis–infected deer to crude mycobacterial antigens. Deer were experimentally inoculated with M. bovis strain 1315 either by intratonsilar instillation or by exposure to M. bovis–infected (i.e., in contact) deer. To determine the time course of the response, including the effects of antigen administration for comparative cervical skin testing, serum was collected periodically and evaluated by enzyme-linked immunosorbent assay (ELISA) for immunoglobulin (i.e., IgG heavy and light chains) reactivity to mycobacterial antigens. The reactivity to M. bovis purified protein derivative (PPDb) exceeded (P < 0.05) the reactivity to M. avium PPD (PPDa) only after in vivo administration of PPDa and PPDb for comparative cervical testing of the infected deer. The mean immunoglobulin response, as measured by ELISA, of intratonsilar-inoculated deer to a proteinase K–digested whole-cell sonicate (WCS-PK) of M. bovis strain 1315 exceeded (P < 0.05) the mean of the prechallenge responses to this antigen at approximately 1 month after inoculation and throughout the remainder of the study (i.e., ~11 months). This response also exceeded (P < 0.05) that of the uninfected deer. Although this is encouraging, further studies are necessary to validate the use of the proteinase K–digested M. bovis antigens in the antibody-based assays of tuberculosis.

Evaluation of an In Vitro Blood-Based Assay to Detect Production of Interferon-γ By <i>Mycobacterium Bovisi>–Infected White-Tailed Deer (<i>Odocoileus Virginianusi>)

Tuberculosis due to Mycobacterium bovis in captive Cervidae was identified as an important disease in the United States in 1990 and prompted the addition of captive Cervidae to the USDA Uniform Methods and Rules for eradication of bovine tuberculosis. As well, M. bovis infection was identified in free-ranging white-tailed deer in northeast Michigan in 1995. Tuberculosis in both captive and free-ranging Cervidae represents a serious challenge to the eradication of M. bovis infection from the United States. Currently, the only approved antemortem tests for tuberculosis in Cervidae are the intradermal tuberculin skin test and the blood tuberculosis test (BTB). At present, the BTB is not available in North America. Tuberculin skin testing of Cervidae is time-consuming and involves repeated animal handling and risk of injury to animals and humans. This study evaluated the potential of a new blood-based assay for tuberculosis in Cervidae that would decrease animal handling, stress, and losses due to injury. In addition, a blood-based assay could provide a more rapid diagnosis. Twenty 6–9-month-old white-tailed deer, male and female, were experimentally inoculated by instillation of 300 colony-forming units of M. bovis in the tonsillar crypts. Seven, age-matched uninfected deer served as controls. Blood was collected on days 90, 126, 158, 180, 210, 238, 263, and 307 after inoculation and was analyzed for the production of interferon-γ (IFN-γ) in response to incubation with M. bovis purified protein derivative (PPDb), M. avium PPDa, pokeweed mitogen (PWM), or media alone. Production of IFN-g in response to PPDb was significantly greater (P < 0.05) at all time points in samples from M. bovis–infected deer as compared with uninfected control deer, whereas IFN-γ production to PWM did not differ significantly between infected and control deer. Measurement of IFN-γ production to PPDb may serve as a useful assay for the antemortem diagnosis of tuberculosis in Cervidae.

The Viability of <i>B. tuberculosis i>(<i>Bovinusi>) on Pasture Land, In Stored Faeces and In Liquid Manure

The object of these experiments was to determine the length of time during which B. tuberculosis in cow's faeces remain alive and virulent on pasture land in the south of England. The method of testing for living B. tuberculosis is given in Appendix II.

Antibody Responses in Reindeer (<i>Rangifer tarandusi>) Infected with <i>Mycobacterium bovisi>

Despite having a very low incidence of disease, reindeer (Rangifer tarandus) are subject to tuberculosis (TB) testing requirements for interstate shipment and herd accreditation in the United States. Improved TB tests are desperately needed, as many reindeer are falsely classified as reactors by current testing procedures. Sera collected sequentially from 11 (experimentally) Mycobacterium bovis-infected reindeer and 4 noninfected reindeer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for antibody specific to M. bovis antigens. Specific antibody was detected as early as 4 weeks after challenge with M. bovis. By MAPIA, sera were tested with 12 native and recombinant antigens, which were used to coat nitrocellulose. All M. bovis-infected reindeer developed responses to MPB83 and a fusion protein, Acr1/MPB83, and 9/11 had responses to MPB70. Other antigens less commonly recognized included MPB59, ESAT-6, and CFP10. Administration of purified protein derivatives for skin testing boosted serum antibody responses, as detected by each of the assays. Of the noninfected reindeer, 2/4 had responses that were detectable immediately following skin testing, which correlated with pathological findings (i.e., presence of granulomatous lesions yet the absence of acid-fast bacteria). The levels of specific antibody produced by infected reindeer appeared to be associated with disease progression but not with cell-mediated immunity. These findings indicate that M. bovis infection of reindeer elicits an antibody response to multiple antigens that can be boosted by skin testing. Serological tests using carefully selected specific antigens have potential for early detection of infections in reindeer.

Constructability Testing of Folded Plate Girders

A new steel girder bridge system was developed at the University of Nebraska. The innovative girder design is a box girder folded from a single steel plate that has a trapezoid shape with an opening on the bottom. The girder has application in short span bridges and accelerated construction situations. The structural performance of the girder requires investigation in all stages of a bridge’s lifecycle. This thesis contains descriptions and results from the first two tests from a series of tests developed to evaluate this new girder shape. The objective of these two tests was to investigate the constructability of the girders. During construction a bridge is in its least stable condition and it is important that the bridge components exhibit both adequate strength and stability during this critical stage. To this end, two girders were tested in flexure over a simple span as a non-composite beam simulating the loading the girders would be subjected to during construction. The results of the two tests indicate that the folded girder as a whole, and its components, provide adequate strength and stability at construction load levels. Failure occurred at loads that were above normal construction load levels and resulted in a ductile failure mode, which is a well documented benefit of steel components. The girders remained stable through all phases of loading including failure. The top flange was the weakest component of the beam during construction due to its role as a compression element that has a slender and un-braced form. The compression in the top flange caused local buckling in the top flange even at elastic load levels. This was the cause for loss of stiffness and failure in both specimens. Incorporation of a ridge at the center of the top flange of specimens, results of which are not reported in this thesis, proved to resolve this very early buckling issue.

Review of Julien Gœury: <i>L 'Autopsie et le théorème: poétique des Théorèmes spirituels (1613-1622) de Jean de La Ceppède.i>

Les lecteurs de La Ceppède seront contents de recevoir le livre de Julien Gœury car son étude représente une addition importante aux ouvrages récents sur le poète aixois. Faisant partie de la redécouverte critique des Théorèmes initiée par les travaux de Jean Rousset dans les années 50, L'Autopsie et le théorème jette un nouveau regard sur l'oeuvre laceppédienne en adoptant ce que l'on peut appeler une perspective néo-structuraliste. L'exposé se divise en quatre parties: 1) Morphologie, 2) Anatomie, 3) Physiologie et 4) Psychologie. Une telle répartition suggere le désir de dégager le caractère organique du texte dans le cadre d'une organisation bien schématisée. Concernant la première categorie, Gœury met en exergue la construction générale du texte, signalant au départ “l'architecture extérieure” (23) ainsi que “l'architecture intérieure” (54) dans la composition des livres et des recueils qui édifient l'ouvrage. Ici, le lecteur note l'accent mis sur la signification du frontispice, des pages de titres, et sur d'autres éléments paratextuels. Toujours dans la première partie, Gœury suit l'exemple de plusieurs critiques en examinant l'emploi du sonnet comme mode de discours. L'auteur met en avant des “lois de composition” (141) qui renforcent “l'engagement formel” (151) du texte ainsi que son “architecture phonetique” (157). S'ajoutent à l'examen morphologique des observations sur les différentes formes “d'enjambement” (168) et de “fragmentation” (174) qui se manifestent dans les sonnets.