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United States Department of Agriculture

Agricultural Research Service

Title: Analysis of Interferon-Gamma Production by Mycobacterium Bovis Infected White-Tailed Deer (Odocoileus Virginianus) Using An in-Vitro Blood Based Assay

Authors
item Palmer, Mitchell
item Waters, Wade
item Whipple, Diana
item Slaughter, Ralph - BIOCOR ANIMAL HEALTH
item Jones, Stephen - CSL LIMITED

Submitted to: Journal of Veterinary Diagnostic Investigation
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 3, 2003
Publication Date: January 16, 2004
Citation: Palmer, M.V., Waters, W.R., Whipple, D.L., Slaughter, R.E., Jones, S.L. 2004. Analysis of interferon-gamma production by mycobacterium bovis infected white-tailed deer (odocoileus virginianus) using an in-vitro blood based assay. Journal of Veterinary Diagnostic Investigation. 16:17-21.

Interpretive Summary: Tuberculosis due to Mycobacterium bovis in captive Cervidae was identified as an important disease in the US in 1990 and prompted the addition of captive Cervidae to the USDA program for the 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 eradication of M. bovis infection from the US. Currently, the only approved test for tuberculosis in Cervidae is tuberculin skin testing. 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. Additionally a blood-based assay could provide a more rapid diagnosis. Experimentally infected white-tailed deer were used while age-matched uninfected deer served as controls. Blood was collected periodically after inoculation and analyzed for the production of the chemical messenger, interferon-gamma (IFN-gamma) Production of IFN-gamma was significantly greater at all time points in samples from M. bovis infected deer as compared to uninfected control deer. Measurement of IFN-gamma production to may serve as a useful assay for the antemortem diagnosis of tuberculosis in Cervidae.

Technical Abstract: Tuberculosis due to Mycobacterium bovis in captive Cervidae was identified as an important disease in the US in 1990 and prompted the addition of captive Cervidae to the USDA Uniform Methods and Rules for the 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 eradication of M. bovis infection from the US. Currently, the only approved test for tuberculosis in Cervidae is intradermal tuberculin skin testing. 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. Additionally 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 CFU 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 analyzed for the production of interferon-gamma (IFN-gamma) in response to incubation with M. bovis PPD, M. avium PPD, pokeweed mitogen (PWM), or media alone. Production of IFN-gamma in response to M. bovis PPD was significantly greater (P<0.05) at all time points in samples from M. bovis infected deer as compared to uninfected control deer. Measurement of IFN-gamma production to M. bovis PPD may serve as a useful assay for the antemortem diagnosis of tuberculosis in Cervidae.

Last Modified: 9/10/2014
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