Tuberculosis in Rhinoceros: An Underrecognized Threat? M. Miller1, A. Michel2, P. van Helden1 and P. Buss3 1DST/NRF Centre of Excellence for Biomedical TB Research/MRC Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, Cape Town, South Africa 2Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa 3Veterinary Wildlife Services, South African National Parks, Kruger National Park, Skukuza, South Africa *Correspondence: M. Miller. PO Box 86, Skukuza 1350, South Africa. Tel.: +27 73 812 9141; E-mail: [email protected] Summary Historical evidence of tuberculosis (TB) affecting primarily captive rhinoceroses dates back almost two centuries. Although the causative Mycobacterium tuberculosis complex (MTC) species has not been determined in many cases, especially for those that occurred before bacterial culture techniques were available, the spectrum of documented reports illustrates the importance of TB as cause of morbidity and mortality in different rhinoceros species across continents. In more recent years, sporadic suspected or confirmed cases of TB caused by Mycobacterium bovis (M. bovis) have been reported in semi-free or free-ranging rhinoceroses in South Africa. However, the true risk TB may pose to the health and conservation of rhinoceros populations in the country's large conservation areas where M. bovis is endemic, which is unknown. Underlying the current knowledge gap is the lack of diagnostic tools available to detect infection in living animals. As documented in other wildlife species, TB could establish itself in a rhinoceros population but remain unrecognized for decades with detrimental implications for wildlife conservation at large and should such animals be moved to uninfected areas or facilities. This paper reviews the current state of knowledge regarding TB in rhinoceros including critical gaps that need to be addressed to effectively assess the threat that this disease may present to rhinoceros. Keywords: Ceratotherium simum; Dicerorhinus sumatrensis; Diceros bicornis; mycobacteriosis; rhinoceros; Rhinoceros unicornis; tuberculosis 1 Introduction Tuberculosis (TB) affects many free-ranging and captive wildlife species and presents a potential threat to conservation efforts. Detection of infection often occurs only after development of chronic, debilitating disease when clinical signs become apparent (Bengis, 1999). Susceptibility to infection, pathogenesis and impact on the affected population differs widely between species, although relatively little is known for most wildlife species (Michel et al., 2006; Pesciaroli et al., 2014). Mycobacterium bovis (M. bovis) and Mycobacterium tuberculosis (M. tuberculosis) have been shown to cause morbidity and mortality in rhinoceros species (Stetter et al., 1995; Rookmaaker et al., 1998; Espie et al., 2009). Rhinoceros are iconic species that have been exported to zoos worldwide and translocated for conservation programmes. However, large knowledge gaps exist in understanding susceptibility, transmission, diagnosis, development of disease and management of infected animals. Although TB has not been shown to be an immediate threat to free-ranging rhinoceros populations, the potential impact is currently unknown. Kruger National Park (KNP) and iMfolozi-Hluhluwe Game Reserves (HiP), KwaZulu Natal, South Africa, have large numbers of both white (Ceratotherium simum) and black (Diceros bicornis) rhinoceros which share range and resources with M. bovis-infected buffalo populations, which are important maintenance hosts for this disease (Michel et al., 2006; Fitzgerald and Kaneene, 2013). A fatal case of M. bovis infection in a recently translocated black rhinoceros on a private game ranch in South Africa (M. Otto, pers. comm. 2014) emphasizes the importance of developing accurate methods of detection suitable for surveillance and early diagnosis. TB in rhinoceros may have other consequences associated with zoonotic risks and regulatory constraints as a notifiable disease. In the event of M. bovis being diagnosed in a rhinoceros, regulatory interventions could result in quarantine of a zoo, park, or game ranch and restrictions placed on further movements of rhinoceros. In response to the inter- national demand for TB diagnostic tests in wildlife, tools such as the STAT-PAK assay (Lyashchenko et al., 2008; Duncan et al., 2009) and rhinoceros-specific interferon-gamma (IFN-gamma) assay have been recently developed (Morar et al., 2013). However, these tests need to be validated in known infected rhinoceros before they can be used reliably for diag- nostic purposes and risk assessments. At the same time, knowledge of the pathogenesis of this multistage disease in rhinoceros is a prerequisite for understanding the risk of contract- ing, developing disease and transmitting the infection. Ultimately, gaining knowledge on a few elementary disease determinants constitutes the key to successfully managing a slow, progressive disease such as TB without over- or under-estimating the necessity for control measures. This study reviews the current state of knowledge of TB both in captive and free- ranging rhinoceros and identifies critical gaps required to address the threat that this disease presents to rhinoceros. Historical Case Reports of TB in Rhinoceros Although there is very limited information for the presence of infection in free-ranging rhinoceros due to logistical and technical constraints, TB in captive rhinoceros held in zoological collections dates back to the late 1800s (Table 1). The popularity of the different 2 rhinoceros species for exhibition led to their distribution worldwide, which often required long transport in close contact with humans. Table 1. Summary of TB cases in rhinoceros Rhinoceros Date Location MTC species Key finding species Sumatran 1879 Zoo – England Unknown Died; TB in lung, spleen rhinoceros 1892 Zoo – Poland Black rhinoceros Unknown Died of TB Circa Sumatran Zoo – India Unknown Died; TB in lung, liver 1900 rhinoceros Sumatran 1910 Zoo – Austria Unknown Died; TB in lung rhinoceros 1941 Zoo – England Indian rhinoceros Unknown Died of TB 1944 Zoo – Germany Black rhinoceros Unknown Died of TB 1956 Zoo – South Africa Black rhinoceros Unknown Died of TB 1957 Zoo – Germany Black rhinoceros Unknown Died of TB 1961 Zoo – Japan Black rhinoceros Unknown Died of TB 1969 Zoo – Czech Republic Black rhinoceros Unknown Died of TB Free-ranging – South 1970 Black rhinoceros M. bovis Africa 1978 Zoo – USA Black rhinoceros M. bovis Ante-mortem diagnosis 1979 Zoo – USA Black rhinoceros M. bovis Ante-mortem diagnosis 1984 Zoo – USA Black rhinoceros Unknown Died of TB Early Free-ranging – South Black rhinoceros Unknown Died; TB lesions 1990s Africa 1990 Zoo – USA Black rhinoceros M. tuberculosis 1991 Zoo – USA White rhinoceros M. bovis Died; pulmonary TB 1992 Zoo – India Black rhinoceros M. tuberculosis Died; pulmonary TB 1994 Zoo – India Black rhinoceros M. tuberculosis Died; pulmonary TB 1994 Zoo – USA Black rhinoceros M. tuberculosis Treated; died Treated; euthanized; negative at 2001 Zoo – USA Black rhinoceros M. tuberculosis necropsy 2002 Zoo – Sweden White rhinoceros M. tuberculosis Euthanized; ZN+ lung granulomas 2007 Zoo – South Africa Black rhinoceros M. bovis Euthanized; incidental lesions 2013 Zoo – USA Black rhinoceros M. tuberculosis Died; pulmonary TB Private reserve – South 2014 Black rhinoceros M. bovis Died; pulmonary TB Africa Sumatran rhinoceros (Dicerorhinus sumatrensis) were periodically displayed in European and Asian zoos around the turn of the 20th century (Rookmaaker et al., 1998). A pair of Sumatran rhinoceros was transported from Malaysia to the Alipore Zoological Gardens, Calcutta, India, and the female died within a year of arrival with TB of the lungs and liver. Pulmonary TB was reported as the cause of death in another female Sumatran rhinoceros housed at the Schönbrunner Tiergarten (Vienna, Austria). The Austrian consul purchased the animal from the Botanical Gardens in Singapore and transported her to Europe where she lived from 1900 to 1910. 3 The London Zoo has had two recorded historical cases of TB in rhinoceros (Rookmaaker et al., 1998). A male Sumatran rhinoceros died shortly after arrival at the zoo in April 1879 with tuberculous lesions in the lungs and spleen. A young male Indian rhinoceros (Rhinoceros unicornis) was transported from Calcutta by ship and arrived at the zoo in 1924. The animal reportedly died of TB in 1941. There are several recorded historical cases in which TB was identified as the cause of death in black rhinoceros housed in zoos (Rookmaaker et al., 1998). Interestingly, all five animals of known gender were males. The earliest report of TB was in a black rhinoceros that died at Miejski Ogrod Zoologiczny (Wroclaw, Poland) in 1892. Prior to arriving at the zoo in 1888, the animal had travelled with Hagenbeck's Nubian Africa show for 10 years. A male black rhinoceros was imported from East Africa to the Zoologischer Garten (Dresden, Germany) in 1928 and died in 1944 from TB. Another male black rhinoceros died in 1957 from TB at Zoologischer Garten Köln (Cologne, Germany) after being resident for 4 years. More recent- ly, an imported male black rhinoceros succumbed to TB at the Zoologicka Zahrada (Prague, Czech Republic) in 1969 after living at the zoo for 15 years. There are also cases of TB in captive black rhinoceros in other parts