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The order Onygenales contains multiple fungal pathogens that affect free-ranging and zoo-housed reptilian species. Emydomyces testavorans, an onygenalean fungus associated with skin and shell disease, has been sporadically detected in aquatic chelonians. Because of the recent discovery of this organism, little is known about its prevalence in free-ranging chelonians. The objective of this study was to perform surveillance for E. testavorans in six free-ranging aquatic and terrestrial chelonian species in Illinois, USA: Blanding's turtles (n=437; Emydoidea blandingii), painted turtles (n=199; Chrysemys picta), common snapping turtles (n=35; Chelydra serpentina), red-eared sliders (n=62; RES; Trachemys scripta elegans), eastern box turtles (n=73; Terrapene carolina carolina) and ornate box turtles (n=29; Terrapene ornata). Combined cloacal-oral swabs (COSs) or shell (carapace and plastron surfaces) swabs were collected from 2019 to 2021 and tested for E. testavorans using quantitative PCR. The PCR detected E. testavorans in COSs of an adult male, subadult female, and juvenile male Blanding's turtle (0.6%; 95% confidence interval [CI], 0.2-1.9%) and a shell swab from an adult female RES (1.6%; 95% CI, 0-8.7%). Shell lesions consistent with E. testavorans infection were present in two of the positive Blanding's turtles. These results document the rarity of this pathogen on the landscape in Illinois. Additional studies should determine this pathogen's impact on individuals and clarify its significance for conservation efforts of Blanding's turtle, in which E. testavorans has not been reported previously. [ABSTRACT FROM AUTHOR]

Two studies have identified Coxiella burnetii in poikilotherms (vertebrates that cannot regulate body temperature physiologically); both studies originated in India. Two tortoises had antibodies to C. burnetii by capillary agglutination [...]

Freshwater turtles face numerous anthropogenic threats worldwide. Health assessments are a key component of chelonian population assessment and monitoring but are under reported in many species. The purpose of this study was to characterize the health of spotted turtles (Clemmys guttata; n = 30) and painted turtles (Chrysemys picta; n = 24) at Camp Edwards, a military base in Cape Cod, Massachusetts, using physical examinations, hematology, plasma heavy metal analyses, and pathogen surveillance via PCR. Spotted turtles had a high prevalence of carapace (n = 27, 90%) and plastron (n = 14, 46.7%) lesions, and a previously undescribed adenovirus was detected in three animals (proposed as Clemmys adenovirus-1). Female painted turtles had lower plasma copper (p = 0.012) and higher strontium (p = 0.0003) than males, and appeared to be in a similar plane of health to previous reports. This initial health assessment effort provides useful baseline data for future comparison in these species. Conservation efforts on Camp Edwards should incorporate continued health surveillance of these populations to identify intervention opportunities and determine the conservation threats, if any, of the novel adenovirus. [ABSTRACT FROM AUTHOR]

Uric acid (UA) is excreted as an end product of protein metabolism in many reptiles, including some chelonians. Elevated plasma UA concentrations can occur due to many physiologic and pathologic changes, and determining plasma UA concentrations is part of a complete general health assessment in this taxon. UA concentrations are typically measured using benchtop chemistry analyzers, but point-of-care (POC) UA meters have also been developed for human use. However, these POC UA meters have not been investigated for use in any reptile species. The purpose of this study was to assess agreement between UA measurements produced by a standard benchtop chemistry analyzer and a POC UA meter in free-living eastern box turtles (Terrapene carolina carolina). UA concentrations were measured with a POC meter using fresh whole blood and frozen-thawed plasma and with a standard benchtop chemistry analyzer using frozen-thawed plasma. Poor-to-moderate agreement was present between each of the three methods as evidenced by mixed models, Passing–Bablok regression, Bland–Altman plots, and Cohen's κ. Differences between methods fell outside of clinically acceptable limits, indicating that the POC UA meter should not be used in eastern box turtles. [ABSTRACT FROM AUTHOR]

Turtles and tortoises are among the most threatened vertebrate groups. Their life history is characterized by delayed sexual maturity and a long lifespan, making populations susceptible to decline following perturbations. Despite the urgent conservation need, we are missing estimates of basic demographic traits for many species and populations. The ornate box turtle Terrapene ornata is a species lacking crucial demographic data. Many populations are isolated in fragmented habitats, especially in the eastern portion of their range. We carried out long-term capture-mark-recapture surveys on two isolated populations in northern Illinois to estimate population vital rates and project population persistence with deterministic stage-based matrix models. Using 34 years of data, we estimated adult female survival = 0.974 (95% CI: 0.946-0.988) and juvenile survival = 0.867 (95% CI: 0.688-0.951) at our most intensively surveyed site. At a second site using eight years of data, we estimated adult female survival = 0.897 (95% CI: 0.783-0.954) and juvenile survival = 0.844 (95% CI: 0.551-0.960). Despite seemingly high annual survival rates, populations declined under population projections using mean vital rates. Population growth was most sensitive to adult survival, with increasing sensitivity under more pessimistic scenarios. Our results highlight the importance of long-term demographic studies for threatened species and demonstrate protecting adult female ornate box turtles is critical for ensuring populations persist at the northern edge of their distribution. [ABSTRACT FROM AUTHOR]

Evaluating leukocytes provides insight in the overall health of animals by characterizing pathologies related to infection, inflammation, and stress. In reptile species, many leukocyte quantification methods are used to evaluate white blood cell (WBC) parameters; however, agreement and analytical variability (repeatability) between the various methods have yet to be reported and critically assessed in many species, including the prairie rattlesnake (Crotalus viridis). The objectives of this study were to 1) test agreement between hematology parameters determined by phloxine-based stain (Eosinophilic leukopet, EO), Natt–Herrick's (NH), and estimate (EST) methods using fresh whole blood and lithium-heparinized blood; 2) determine the agreement between estimates from a veterinary student and from a board-certified clinical pathologist; and 3) determine the analytical variability of EO, NH, and EST methods. Venipuncture was performed in 27 adult snakes, and agreement was then assessed using Bland–Altman plots and Passing–Bablock regression between all methods. There was a variable degree of proportional and constant error between many of the quantification methods, but descriptive statistics for all methods fell within biologically relevant values. There was good agreement in the EST method between the clinical pathologist and the veterinary student, but the student had a significantly higher mean WBC count in fresh whole blood. The lowest analytical variability was observed with the NH method (15.4%), whereas the highest was observed with the EST method (58.6% for the clinical pathologist, 59.4% for the veterinary student). These results identify the intrinsic variability in reptile hematological methods currently used and emphasize the need to develop a singular diagnostic standard for leukocyte quantification in reptile species. [ABSTRACT FROM AUTHOR]

Frog virus 3 (FV3) and related ranaviruses are emerging infectious disease threats to ectothermic vertebrate species globally. Although the impact of these viruses on amphibian health is relatively well studied, less is understood about their effects on reptile health. We report two cases of FV3 infection, 11 mo apart, in three-toed box turtles (Terrapene mexicana triunguis) from a wildlife rehabilitation center. Case 1 had upper respiratory signs upon intake but had no clinical signs at the time of euthanasia 1 mo later. Case 2 presented for vehicular trauma, had ulcerative pharyngitis and glossitis, and died overnight. In case 1, we detected FV3 nucleic acid with qPCR in oral swabs, kidney, liver, spleen, and tongue. In case 2, we detected FV3 in an oral swab, an oral plaque, heart, kidney, lung, liver, spleen, and tongue. We also detected FV3 nucleic acid with in situ hybridization for case 2. For both cases, FV3 was isolated in cell culture and identified with DNA sequencing. Histopathologic examination of postmortem tissue from case 1 was unremarkable, whereas acute hemorrhagic pneumonia and splenic necrosis were noted in case 2. The difference in clinical signs between the two cases may have been due to differences in the temporal course of FV3 disease at the time of necropsy. Failure to detect this infection previously in Missouri reptiles may be due to lack of surveillance, although cases may also represent a novel spillover to box turtles in Missouri. Our findings reiterate previous suggestions that the range of FV3 infection may be greater than previously documented and that infection may occur in host species yet to be tested. [ABSTRACT FROM AUTHOR]

Eastern box turtles (Terrapene carolina carolina) face a variety of anthropogenic, infectious, and environmental threats and have been affected by high morbidity and mortality disease events. Wellness parameters in free-ranging eastern box turtles with a high prevalence of myiasis on Cape Cod, MA, were documented to identify epidemiologic trends or associations with several health parameters. There were 109 samples collected from 59 individual box turtles over the course of 4 mon. Six turtles died over the course of this study. Fly larvae infestations varied in severity and were observed in the cutaneous and subcutaneous tissue (n = 18; 30.5%). Animals with myiasis had fewer plastron abnormalities than those without (P = 0.034), and all turtles found in bogs had evidence of fly larvae infections (P < 0.0001). Individuals with myiasis also had lower body condition index (P = 0.014), lower total white blood cells (P = 0.031), lower PCV (P < 0.0001), lower total solids (P < 0.0001), higher erythrocyte sedimentation rate (P < 0.0001), lower calcium (P = 0.018), and lower phosphorus (P = 0.017). Three turtles tested positive for terrapene herpesvirus 1, but presence was not associated with myiasis. Heavy metal analysis revealed no significant differences between turtles with and without myiasis. This study examined the health of a population of eastern box turtles, and continued health assessments will be beneficial in determining the impact of myiasis on future conservation plans. [ABSTRACT FROM AUTHOR]

Anticoagulants prevent clotting of blood samples and preserve cellular morphology for hematologic evaluations, but studies comparing anticoagulants are limited in snakes. The objective of this study was to evaluate the effects of lithium heparin (LH) and dipotassium ethylenediaminetetraacetic acid (EDTA) on hematologic values in prairie rattlesnakes (PR; Crotalus viridis, n = 16) and Lake Erie water snakes (LEWS; Nerodia sipedon insularum, n = 21). Venipuncture was performed and blood samples were immediately aliquoted into LH and EDTA microtainers. Packed cell volume (PCV), total solids (TS), 100-cell differential counts, and Avian Leukopet white blood cell counts (WBC) were determined for each anticoagulant. Passing–Bablok regression and Bland–Altman plots revealed that anticoagulant choice did not constantly or proportionally bias the values of any WBC parameter. Mixed models demonstrated that blood anticoagulated with EDTA had higher PCV in PR (P = 0.04) and TS in both species (P < 0.05). However, the magnitude of the differences attributable to anticoagulant choice was relatively small and likely not clinically important. Hemolysis was not appreciated in any samples. Our findings demonstrate that LH and EDTA are equally appropriate for use in PR and LEWS, but may require separate reference values. [ABSTRACT FROM AUTHOR]

Freshwater turtles are sensitive to environmental changes and can serve as sentinel species for ecosystem health assessments. The Efroymson Restoration at Kankakee Sands in northwestern Indiana, USA has been restored in the past 25 yr from primarily agricultural land to a mosaic of prairie and wetland habitats. Health assessments of 40 free-ranging painted turtles (Chrysemys picta) at Kankakee Sands were performed in May 2021 to evaluate overall health status, screen for infectious diseases, and obtain baseline clinical pathology values in this population. Assessment of each turtle included a physical examination, complete blood count, plasma biochemistry panel, blood lactate level, venous blood gas analysis, serum trace mineral panel, serum vitamin D3 level, and plasma protein electrophoresis. Oral and cloacal swabs were tested for adenoviruses, herpesviruses, frog virus 3, and Mycoplasmopsis species by PCR in 39 painted turtles. Four turtles were positive for adenovirus, which shared 100% homology to Sulawesi tortoise adenovirus. Two turtles were herpesvirus-positive with 100% homology to emydid herpesvirus 1. No Mycoplasmopsis spp. or frog virus 3 was detected. Female turtles had significantly higher manganese, prealbumin, uric acid, triglycerides, and ionized calcium levels, while male turtles had significantly higher cholesterol, glutamate dehydrogenase, and CO2 levels. These baseline data can be used for future research into freshwater turtle health in restored wetland habitats. [ABSTRACT FROM AUTHOR]

Raccoon roundworm (Baylisascaris procyonis) negatively affects woodrat (Neotoma spp.) populations but is not known to occur in the endemic range of endangered Key Largo woodrats (Neotoma floridana smalli). Rectal swabs from 23 raccoons (Procyon lotor) in Key Largo were screened for raccoon roundworm by PCR. All tests were negative, suggesting continued absence. [ABSTRACT FROM AUTHOR]