Novel Assays for Detection of Mycobacterium bovis Infection in Free-Ranging African Rhinoceros (Diceros bicornis, Ceratotherium simum) and Implications for Conservation
2018 Joint EAZWV/AAZV/Leibniz-IZW Conference
Michele A. Miller1, DVM, MPH, PhD, DECZM (ZHM); Peter E. Buss2, BVSc, MMedVet, PhD; Josephine Chileshe1, BSc (Hons), MSc; Wynand Goosen1, BSc (Hons), PhD; Eduard Roos1, BSc (Hons); Paul van Helden1, PhD; Sven D.C. Parsons1, BVSc, PhD
1Department of Science and Technology/National Research Foundation Centre of Excellence for Biomedical TB Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, Cape Town, South Africa; 2Veterinary Wildlife Services, South African National Parks, Kruger National Park, Skukuza, South Africa


Tuberculosis (TB) in captive African rhinoceros has been sporadically reported.2 However, mycobacterial infection has only recently been observed in rhinoceros in range countries.1,3,5 Between 2016 and 2018, Mycobacterium bovis (M. bovis) infection was confirmed in one black and five white rhinoceros in Kruger National Park, South Africa. The South African Department of Agriculture, Forest, and Fisheries (DAFF) issued a quarantine notice in December 2016, which has prevented further rhinoceros translocations until a management plan could be created that included TB testing of rhinoceros. A novel interferon-gamma (IFN-γ) release assay (IGRA), based on the human QuantiFERON® TB Gold In-Tube (QFT) (Qiagen, Hilden, Germany) whole blood stimulation platform and a commercially available equine IFN-γ ELISA (Mabtech AB, Nacka Strand, Sweden) for cytokine detection, was developed and validated for use in black and white rhinoceros. Samples from known M. bovis infected and uninfected white rhinoceros were used to determine response kinetics and diagnostic cut-off value for the ELISA.4 Screening of free-ranging and boma-confined rhinoceros has detected individuals that have had consistent positive IGRA results on serial samples, suggesting that these may be infected and require further confirmatory testing. These preliminary findings indicate that this IGRA, using commercially available reagents, has promise as diagnostic tool to screen African rhinoceros for mycobacterial infection and facilitate future translocations, especially from TB endemic populations.


The authors would like to thank the following the individuals for their contributions to this study: Leana Rossouw, Guy Hausler, Tebogo Manamela, Dr. Markus Hofmeyr, Dr. Lin-Mari deKlerk-Lorist, Dr. Louis van Schalkwyk, and the Veterinary Wildlife Services capture team from Kruger National Park for assistance with sample collection from the rhinoceros. The authors also acknowledge Drs. Eva Gelius and Jens Gertow of Mabtech Ab for their assistance. This work was supported by the South African Medical Research Council, the National Research Foundation of South Africa [SARChI grant 86949], and the American Association of Zoo Veterinarians Wild Animal Health Fund. The content is the sole responsibility of the authors and does not necessarily represent the official views of the funders.

Literature Cited

1.  Espie IW, Hlokwe TM, van Pittius NCG, Lane E, Tordiffe AS, Michel AL, Müller A, Kotze A, Van Helden PD. Pulmonary infection due to Mycobacterium bovis in a black rhinoceros (Diceros bicornis minor) in South Africa. J Wildl Dis. 2009;45:1187–1193.

2.  Miller M, Michel A, van Helden P, Buss P. Tuberculosis in rhinoceros: an underrecognized threat? Transbound Emerg Dis. 2016;doi:10.1111/tbed.12489.

3.  Miller M, Buss P, van Helden P, Parsons S. Mycobacterium bovis—report of tuberculosis in a free-ranging black rhinoceros (Diceros bicornis) in Kruger National Park, South Africa 2016. Emerg Inf Dis. 2017:23:557–558.

4.  Parsons SD, Morar-Leather D, Buss P, Hofmeyr J, McFadyen R, Rutten VP, van Helden P, Miller MA, Michel AL. The kinetics of the humoral and interferon-gamma immune responses to experimental Mycobacterium bovis infection in the white rhinoceros (Ceratotherium simum). Front Immunol. 2017;doi: 10.3389/fimmu.2017.01831.

5.  Thapa J, Paudel S, Sadaula A, Shah Y, Maharjan B, Kaufman GE, McCauley D, Gairhe KP, Tsubota T, Suzuki Y, Nakajima C. Mycobacterium orygis-associated tuberculosis in free-ranging rhinoceros, Nepal, 2015. Emerg Infect Dis. 2016;22:570.


Speaker Information
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Michele A. Miller, DVM, MPH, PhD, DECZM (ZHM)
Department of Science and Technology
Stellenbosch University
Cape Town, South Africa

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