Mycoplasmopsis-associated Proliferative Pneumonia in a Bog Turtle (Glyptemys muhlenbergii).

Lower respiratory tract disease associated with mycoplasmal infection was detected in a free-ranging bog turtle (Glyptemys muhlenbergii) from New Jersey, US. The presence of a mycoplasmal organism was confirmed by PCR and electron microscopy. Fluid-filled lungs were observed grossly, and there was proliferative pneumonia on histopathology. Respiratory disease caused by Mycoplasmopsis (Mycoplasma) spp. has been widely documented across animal taxa. In reptiles, these infections are predominantly implicated in upper respiratory tract disease (URTD). Typical disease in chelonids presents as oculonasal discharge, conjunctivitis, palpebral edema, and rhinitis, which is most frequently associated with Mycoplasma agassizii and Mycoplasma testudineum and is largely identified in tortoises (Kolesnik et al. 2017; Pasmans et al. 2021). Mycoplasmosis is reported less frequently in turtles, but it has been associated with URTD in Eastern box turtles (Terrapene carolina carolina; Pasmans et al. 2021) and European pond turtles (Emys orbicularis; Schönbächler et al. 2022) and documented in European diagnostic submission surveys in turtles from the Emydidae, Geoemydidae, Kinosternidae, and Chelidae families (Kolesnik et al. 2017). Mycoplasma spp. have also been identified in the absence of clinical disease in multiple species, including North American western pond turtles (Actinemys [Emys] marmorata), red-eared sliders (Trachemys scripta elegans; Silbernagel et al. 2013), three-toed box turtles (Terrapene carolina triunguis; Palmer et al. 2016), spotted turtles (Clemmys guttata), and bog turtles (Glyptemys muhlenbergii; Ossiboff et al. 2015). In contrast, documented reports of lower respiratory tract disease in reptiles with mycoplasmosis are scant. A single case of proliferative tracheitis and pneumonia in a Burmese python (Python molurus bivittatus) was associated with a novel Mycoplasma sp. (Penner et al. 1997).

HEALTH ASSESSMENT OF WILD NORTHERN DIAMOND-BACKED TERRAPINS (MALACLEMYS TERRAPIN TERRAPIN) IN NEW JERSEY, USA.

A popular species in the pet trade, and therefore in the illegal wildlife trade, the diamond-backed terrapin, Malaclemys terrapin terrapin, population has seen significant declines. Associated with the illegal wildlife trade, occasions arise in which terrapins are confiscated, and no standard operating procedures exist for repatriation into the wild. To develop such procedures, an understanding of the pathogens circulating in the wild diamond-backed terrapin population in New Jersey is needed. We sampled 30 wild female diamond-backed terrapins for herpesvirus, Mycoplasmopsis, ranavirus, and intestinal and blood parasites and performed white blood cell counts and differentials and evaluated biochemistry values. Terrapins had an average age of 10 yr (8-15 yr), and 70% were gravid at the time of sampling. Thirty-three percent of the sampled northern diamond-backed terrapins were positive for Mycoplasmopsis sp., and all were negative for ranavirus and herpesviruses. Occasional blood parasites were found, and few intestinal parasites were noted. There was no significant difference between gravid status and any of the blood parameters (P<0.05). Blood chemistry values appeared to vary according to feeding activity; no differences were noted in the values in relation to gravid status. Four terrapins had heterophil to lymphocyte (H:L) ratios above 4.5, significantly higher than the other terrapins sampled, which may indicate inflammation. Two of the four had Mycoplasmopsis, one sample was contaminated by other bacteria and was discarded, and one was negative. No significant difference was found between Mycoplasmopsis infection status and H:L ratio (P=0.926). Our findings, though conducted on a small number of female terrapins at a specific time point, provide data on the pathogens that may be circulating in this population, adding to the current body of knowledge and helping to guide decision making for the reintroduction of confiscated diamond-backed terrapins into New Jersey's wild population.

Is the future female for turtles? Climate change and wetland configuration predict sex ratios of a freshwater species.

Climate change and land-use change are leading drivers of biodiversity decline, affecting demographic parameters that are important for population persistence. For example, scientists have speculated for decades that climate change may skew adult sex ratios in taxa that express temperature-dependent sex determination (TSD), but limited evidence exists that this phenomenon is occurring in natural settings. For species that are vulnerable to anthropogenic land-use practices, differential mortality among sexes may also skew sex ratios. We sampled the spotted turtle (Clemmys guttata), a freshwater species with TSD, across a large portion of its geographic range (Florida to Maine), to assess the environmental factors influencing adult sex ratios. We present evidence that suggests recent climate change has potentially skewed the adult sex ratio of spotted turtles, with samples following a pattern of increased proportions of females concomitant with warming trends, but only within the warmer areas sampled. At intermediate temperatures, there was no relationship with climate, while in the cooler areas we found the opposite pattern, with samples becoming more male biased with increasing temperatures. These patterns might be explained in part by variation in relative adaptive capacity via phenotypic plasticity in nest site selection. Our findings also suggest that spotted turtles have a context-dependent and multi-scale relationship with land use. We observed a negative relationship between male proportion and the amount of crop cover (within 300 m) when wetlands were less spatially aggregated. However, when wetlands were aggregated, sex ratios remained consistent. This pattern may reflect sex-specific patterns in movement that render males more vulnerable to mortality from agricultural machinery and other threats. Our findings highlight the complexity of species' responses to both climate change and land use, and emphasize the role that landscape structure can play in shaping wildlife population demographics.

Follow-up ecological studies for cryptic species discoveries: Decrypting the leopard frogs of the eastern U.S.

Cryptic species are a challenge for systematics, but their elucidation also may leave critical information gaps about the distribution, conservation status, and behavior of affected species. We use the leopard frogs of the eastern U.S. as a case study of this issue. We refined the known range of the recently described Rana kauffeldi, the Atlantic Coast Leopard Frog, relative to the region's two other leopard frog species, conducted assessments of conservation status, and improved methods for separating the three species using morphological field characters. We conducted over 2,000 call and visual surveys and took photographs of and tissue samples from hundreds of frogs. Genetic analysis supported a three-species taxonomy and provided determinations for 220 individual photographed frogs. Rana kauffeldi was confirmed in eight U.S. states, from North Carolina to southern Connecticut, hewing closely to the Atlantic Coastal Plain. It can be reliably differentiated in life from R. pipiens, and from R. sphenocephala 90% of the time, based on such characters as the femoral reticulum patterning, dorsal spot size and number, and presence of a snout spot. However, the only diagnostic character separating R. kauffeldi from R. sphenocephala remains the breeding call described in 2014. Based on our field study, museum specimens, and prior survey data, we suggest that R. kauffeldi has declined substantially in the northern part of its range, but is more secure in the core of its range. We also report, for the first time, apparent extirpations of R. pipiens from the southeastern portion of its range, previously overlooked because of confusion with R. kauffeldi. We conclude with a generalized ecological research agenda for cryptic species. For R. kauffeldi, needs include descriptions of earlier life stages, studies of niche partitioning with sympatric congeners and the potential for hybridization, and identification of conservation actions to prevent further declines.

Three novel herpesviruses of endangered Clemmys and Glyptemys turtles.

The rich diversity of the world's reptiles is at risk due to significant population declines of broad taxonomic and geographic scope. Significant factors attributed to these declines include habitat loss, pollution, unsustainable collection and infectious disease. To investigate the presence and significance of a potential pathogen on populations of critically endangered bog turtles (Glyptemys muhlenbergii) as well sympatric endangered wood (G. insculpta) and endangered spotted (Clemmys guttata) turtles in the northeastern United States, choanal and cloacal swabs collected from 230 turtles from 19 sites in 5 states were screened for herpesvirus by polymerase chain reaction. We found a high incidence of herpesvirus infection in bog turtles (51.5%; 105/204) and smaller numbers of positive wood (5) and spotted (1) turtles. Sequence and phylogenetic analysis revealed three previously uncharacterized alphaherpesviruses. Glyptemys herpesvirus 1 was the predominant herpesvirus detected and was found exclusively in bog turtles in all states sampled. Glyptemys herpesvirus 2 was found only in wood turtles. Emydid herpesvirus 2 was found in a small number of bog turtles and a single spotted turtle from one state. Based on these findings, Glyptemys herpesvirus 1 appears to be a common infection in the study population, whereas Glyptemys herpesvirus 2 and Emydid herpesvirus 2 were not as frequently detected. Emydid herpesvirus 2 was the only virus detected in more than one species. Herpesviruses are most often associated with subclinical or mild infections in their natural hosts, and no sampled turtles showed overt signs of disease at sampling. However, infection of host-adapted viruses in closely related species can result in significant disease. The pathogenic potential of these viruses, particularly Emydid herpesvirus 2, in sympatric chelonians warrants additional study in order to better understand the relationship of these viruses with their endangered hosts.

A Mycoplasma species of Emydidae turtles in the northeastern USA.

Mycoplasma infections can cause significant morbidity and mortality in captive and wild chelonians. As part of a health assessment of endangered bog turtles (Glyptemys muhlenbergii) in the northeastern US, choanal and cloacal swabs from these and other sympatric species, including spotted turtles (Clemmys guttata), eastern box turtles (Terrapene carolina carolina), wood turtles (Glyptemys insculpta), and common snapping turtles (Chelydra serpentina) from 10 sampling sites in the states (US) of Delaware, New Jersey, and Pennsylvania, were tested by PCR for Mycoplasma. Of 108 turtles tested, 63 (58.3%) were PCR positive for Mycoplasma including 58 of 83 bog turtles (70%), three of three (100%) eastern box turtles, and two of 11 (18%) spotted turtles; all snapping turtles (n = 7) and wood turtles (n = 4) were negative. Sequence analysis of portions of the 16S-23S intergenic spacer region and the 16S ribosomal RNA gene revealed a single, unclassified species of Mycoplasma that has been previously reported in eastern box turtles, ornate box turtles (Terrapene ornata ornata), western pond turtles (Emys marmorata), and red-eared sliders (Trachemys scripta elegans). We document a high incidence of Mycoplasma, in the absence of clinical disease, in wild emydid turtles. These findings, along with wide distribution of the identified Mycoplasma sp. across a broad geographic region, suggest this bacterium is likely a commensal inhabitant of bog turtles, and possibly other species of emydid turtles, in the northeastern US.

Cryptic diversity in metropolis: confirmation of a new leopard frog species (Anura: Ranidae) from New York City and surrounding Atlantic coast regions.

We describe a new cryptic species of leopard frog from the New York City metropolitan area and surrounding coastal regions. This species is morphologically similar to two largely parapatric eastern congeners, Rana sphenocephala and R. pipiens. We primarily use bioacoustic and molecular data to characterize the new species, but also examine other lines of evidence. This discovery is unexpected in one of the largest and most densely populated urban parts of the world. It also demonstrates that new vertebrate species can still be found periodically even in well-studied locales rarely associated with undocumented biodiversity. The new species typically occurs in expansive open-canopied wetlands interspersed with upland patches, but centuries of loss and impact to these habitats give some cause for conservation concern. Other concerns include regional extirpations, fragmented extant populations, and a restricted overall geographic distribution. We assign a type locality within New York City and report a narrow and largely coastal lowland distribution from central Connecticut to northern New Jersey (based on genetic data) and south to North Carolina (based on call data).

Statewide mapping and assessment of vernal pools: a New Jersey case study.

In 2001, the New Jersey Department of Environmental Protection (NJDEP) adopted rules specifically protecting vernal pool habitat for the first time. Vernal pools are small isolated temporary bodies of water that provide critical breeding habitat for a number of amphibian species. To implement these rules and ultimately afford vernal pools protection, the NJDEP first needed to assemble a statewide database of vernal pool locations. In response, the Rutgers University Center for Remote Sensing and Spatial Analysis (CRSSA) was funded to develop a cost effective technique to map vernal pool locations statewide. The objective of CRSSA's mapping effort was to develop a complete potential vernal pool database to be able to identify individual isolated vernal pools as well as areas of high local density, or 'hotspots'. CRSSA used visual interpretation of leaf-off color infrared digital orthophotography in a computerized GIS environment to identify and map over 13,000 potential vernal pools. Using the 1m scale imagery, we determined the minimum detectable pool size to be on the order of 0.02 ha in size. Subsequent field checking has revealed a 12% error of commission that was due to our inclination towards erring on the side of inclusion in mapping many water features as potential vernal pools. For a vernal pool to receive regulated protection, it must be 'certified' that it serves as habitat for obligate or facultative vernal pool amphibian species. To aid in these efforts, CRSSA developed an interactive internet mapping site to assist NJDEP and its citizen volunteer corps in locating and navigating to their survey areas and to facilitate the on-line submittal of survey observations.