Herpetological Review 49(1), 2018 AMPHIBIAN AND REPTILE DISEASES 47 Herpetological Review, 2018, 49(1), 47–49. © 2018 by Society for the Study of Amphibians and Reptiles Seroprevalence of Mycoplasma agassizii and Mycoplasma testudineum in Wild and Waif Gopher Tortoises (Gopherus polyphemus) in Miami-Dade County, Florida, USA Gopher Tortoises (Gopherus polyphemus) are subject to a variety of natural and anthropogenic stressors including habitat loss and degradation, chemical contaminants, overexploitation, and diseases (Rostal et al. 2014). Risk assessment and abatement is especially critical for this species because Gopher Tortoises are classified as a candidate species (Eastern population, including all of Florida, USA) for listing as Threatened under the US Endangered Species Act (USFWS 2014), Florida State Threatened (FWC 2013), and globally Vulnerable by the International Union for the Conservation of Nature Red List (Tortoise & Freshwater Turtle Specialist Group 1996). In particular, Gopher Tortoises are increasingly threatened by emerging infectious diseases (Berish et al. 2010; Jacobson et al. 2014), including Upper Respiratory Tract Disease (hereafter, “URTD”), an infection typically caused by Mycoplasma testudineum or M. agassizii. URTD has been detected in populations of Gopher Tortoises throughout much of Florida (Berish et al. 2000, 2010), and is a health concern both for individual Gopher Tortoises and entire populations (Jacobson et al. 2014; McGuire et al. 2014). In some cases, URTD may cause high mortality and morbidity in Gopher Tortoise populations, with up to 50% of individuals in a population dying in a single acute outbreak (Jacobson et al. 2014). However, infection by Mycoplasma is not necessarily associated with symptoms of disease. Herein, we report seroprevalence for wild and waif Gopher Tortoises from Miami-Dade County, Florida. Our aim is to expand surveillance of tortoise Mycoplasma exposure to the southern end of the species’ distribution, and to provide information relevant for management of the species within highly urbanized environments. We sampled wild and waif Gopher Tortoises for Mycoplasma testudineum and M. agassizii within Miami-Dade County, between September 2015 and April 2017. Our sampling of wild tortoises focused on the Richmond Tract—an isolated 4.5-km 2 patch of critically endangered pine rocklands habitat (Diamond and Heinen 2014; Possley et al. 2016) that harbors one of the few Gopher Tortoise populations within Miami-Dade County and extreme southern Florida (Kushlan and Mazotti 1984; Enge et al. 2004; Rostal et al. 2014). The Richmond Tract consists of several contiguous parcels (including Zoo Miami, Larry & Penny Thompson Memorial Park, Martinez Pineland Preserve, Gold Coast Railroad Museum and several other private and government landowners). We located tortoises sampled in this study by opportunistic encounters of tortoises, and through monitoring apparently active burrows using camera traps and Havahart traps at burrow entrances to capture tortoises from within occupied burrows. In addition to sampling within the Richmond Tract, we also opportunistically sampled two waif tortoises that were brought to Zoo Miami by local residents. Here, we define “waif” tortoise as any tortoise of unknown geographic origin found outside of a natural area or one that had been held for some time in captivity. Waif tortoises are of particular interest because long-distance movement of tortoises may be an important route for pathogen transmission between isolated populations, and with the uncertain origin of waifs, they may represent these types of vectors. In contrast to waif tortoises, we define “wild” tortoises as any tortoise found within a natural area at the Richmond Tract, although it is possible that some tortoises within the Richmond Tract were at some point displaced by people. As one of the few large protected areas of natural habitat within urbanized Miami-Dade County, the Richmond Tract properties are a popular site for illegal release of unwanted exotic pets by Miami-Dade residents (Whitfield, pers. observ.). All tortoises sampled in this study were adults or sub-adults. To test for exposure to pathogens, we extracted up to 2-mL blood from the brachial vein of tortoises. We held plasma or serum from blood draws at -20°C until processing. Samples were analyzed by the University of Florida Tortoise Mycoplasma Testing Lab using enzyme-linked immunoassay tests (ELISA) for M. testudineum and M. agassizii (Wendland et al. 2007). Exposure to M. testudineum was supported for 2 of 12 (17%) tortoises and another two tortoises showed suspect ELISA results (Table 1). Exposure to M. agassizii was supported in 1 of 12 (8%) tortoises and a further two tortoises showed suspect ELISA results for exposure to this pathogen (Table 1). Two tortoises tested positive for exposure to M. agassizii and suspect for M. testudineum. None of the tortoises examined showed clinical symptoms of URTD (e.g., dyspnea, rhinorrhea, epiphora, mucopurulent ocular and/or nasal discharge, conjunctivitis). We are not aware of any reports of disease-associated mortality in Gopher Tortoises in Miami-Dade County. Individual wild tortoises included individuals with both exposure to M. agassizii and M. testudineum, while neither of the two waif tortoises showed exposure to M. agassizii and only one of two tested suspect for exposure to M. testudineum. The ELISA assays for M. agassizii and M. testudineum indicated exposure to a pathogen, not the sustained presence of the pathogen in the Gopher Tortoise population within the Richmond Tract. The sample sizes in this study were low, in particular for waif tortoises. Small sample sizes limit our ability STEVEN M. WHITFIELD Zoo Miami, Conservation and Research Department, Miami, Florida 33177, USA; e-mail: steven.whitfield@miamidade.gov FRANK N. RIDGLEY Zoo Miami, Conservation and Research Department, Miami, Florida 33177, USA DANIEL VALLE Florida International University, Department of Earth and Environment, Miami, Florida 33199, USA NICOLE ATTEBERRY Zoo Miami, Department of Animal Sciences, Miami, Florida 33177, USA