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Zoological institutions manage animals for conservation, education, entertainment, and research purposes. Zoological staff have a responsibility to safeguard the welfare of animals in their care. Retrospective morbidity and/or mortality studies (MMSs) can be useful tools to highlight common diseases in captive wildlife populations. There is currently no standardized methodology for conducting MMSs. Variation in the methodology of MMSs, particularly the categorization of diseases, can make comparisons between studies challenging and may limit the applicability of the results. A Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) compliant systematic review was performed, which identified 67 MMSs describing 146 species of captive wildlife. These MMSs are becoming more common and were predominantly performed on mammals (76/146). Prospective authors are encouraged to perform MMSs on amphibians, birds, reptiles, fish, and invertebrates. The studied animals were mostly managed at institutions in the United States of America (28/67, 41.2%) and Europe (14/67, 20.9%). Classifying individuals into age groups facilitates the identification of disease trends within age classes. Only 22/67 (32.8%) studies cited justification for their age classification; classifications should be based on a referenced source on the breeding biology of the studied species. There is variation in the body systems used by authors and into which system a disease is categorized, which makes study comparisons challenging. Diseases were predominantly categorized by etiology and body system (28/77, 36.4%). Because of its ubiquity, the use of the categorization system employed by the pathology module of the Zoological Information Management System is recommended as a useful standard. This system is imperfect, and amendments to it are suggested. The results and recommendations of this study were discussed with a panel of zoo and wildlife experts; guidelines have been formulated for prospective authors aiming to conduct MMSs in captive wildlife.
Identifying common causes of mortality in zoo giraffe (Giraffa spp.) and okapi (Okapia johnstoni) provides an opportunity to help improve welfare and population management for these endangered species. Mortality reports from 1,024 giraffe and 95 okapi in zoos were compiled from the Species 360 Zoological Information Management Software (ZIMS) utilizing the Morbidity & Mortality Analysis tool. Thirty years of mortality reports (1991–2020) were evaluated to help identify trends and evaluate the impacts, if any, of changes over time in husbandry and management practices. The most common causes of death for giraffe from 1991 to 2015 were neonatal issues (234/845, 27.7%), trauma (213/845, 25.2%), noninfectious disease (190/845, 22.5%), and infectious disease (188/845, 22.2%). In comparison, the most common causes of mortality for giraffe from 2016 to 2020, were noninfectious disease (78/179, 43.6%), trauma (39/179, 21.8%), neonatal issues (39/179, 21.8%), and infectious disease (17/179, 9.5%). The most common cause of death for okapi from 1991 to 2015 were neonatal issues (29/64, 45.3%), infectious disease (13/64, 20.3%), noninfectious disease (11/64, 17.2%), and trauma (10/64, 15.6%). In comparison, the most common cause of death for okapi from 2016 to 2020 was noninfectious disease (15/31, 48.4%), neonatal issues (8/31, 25.8%), and infectious disease (5/31, 16.1%). The results suggest that zoo giraffids have had a relative decrease in mortality from infectious diseases in recent years, whereas death from noninfectious causes has increased significantly. Trauma-related giraffe mortalities and neonatal mortality in both giraffe and okapi, although decreasing in prevalence between time periods, continue to be important causes of death in zoos. This is the first descriptive mortality review for the Giraffidae family and provides data on potential giraffe and okapi health issues that zoos could proactively address.
Insects are increasingly common in households, zoological collections, research, and food industries. Increased knowledge of insect health parameters is necessary to ensure proper welfare. The study goal was to develop a hemolymph collection technique and report hemolymph serum biochemical parameters for the thorny devil stick insect (Eurycantha calcarata). Clinically healthy adult stick insects (5 males and 14 females, 15.1–24.7 g) were enrolled. Four collection techniques were evaluated. Hemolymph collection was unsuccessful in males, but was successful in females by using a single technique. The insect was manually restrained in an elevated position and an 18-ga × 2.54-cm needle was used to puncture the membrane just caudal to the third pair of legs. With the puncture site directed ventrally, ≤1 ml of hemolymph was collected via gravity and allowed to visibly clot. The sample was then centrifuged, and the serum was separated. Serum samples were individually analyzed (Avian/Reptilian Profile Plus, VetScan VS2, Abaxis, Inc, Union City, CA 94587, USA). Fourteen samples (0.2–1.0 ml) were collected from 14 females. Median (minimum–maximum) parameters included the following: aspartate aminotransferase 12 (0–45) U/L, creatinine kinase 25 (0–76) U/L, uric acid 7.5 (3.1–13.7) mg/dl, glucose 12 (8–22) mg/dl, calcium 18.6 (17.2–19.4) mg/dl, phosphorus 15.0 (n = 1) or >30.0 (n = 13) mg/dL, total protein 2.7 (1.6–2.9) g/dL, albumin 0.9 (0.2–1.2) g/dL, globulin 1.7 (1.6–1.8) g/dL (n = 6) or not quantified (n = 8), potassium 10.6 (9.0–11.8) mmol/L, sodium < 100 mmol/L, and bile acids 0 lmol/L. This is the first report of biochemistry parameters in clinically healthy female stick insects. Larger sample sizes are needed to establish statistically valid reference ranges. Hemolymph collection techniques for male stick insects warrant further investigation.
Edmund J. Flach, Mark F. Stidworthy, Sam Aberdeen, David Clarke, Hannah Davidson, Helen Donald, Grace Goodey, Alysa Hulbert, Shinto John, Nic Masters, Shaheed Macgregor, Paul Pearce-Kelly, Simon Spiro, Amanda Guthrie
Between 2015 and 2019, a health screening was carried out annually on captive-bred Partula snails prior to export for reintroduction as part of an international effort to repopulate areas of French Polynesia, where the snails were extinct or critically endangered. In total, 129 separate tank populations of 12 different species were screened at ZSL London Zoo. Wet mounts and smears stained with modified Ziehl–Neelsen (MZN) of 535 fecal samples were examined, and 45% contained flagellated protozoa, and 35.5% had MZN-positive oocysts, measuring 3–5 µm in diameter. Smaller (2 µm) presumptive spores, MZN-positive bacilli, ciliated protozoa and nematodes were recorded less frequently. Fecal bacterial culture yielded mixed species, with a clear predominance of Myroides species (88.9% of samples). The MZN-positive oocysts (3–5 µm) were present in 6.5% of impression smears from the apices of 432 snails examined postmortem, plus acid-fast bacilli in a few cases, but no 2 µm spores. Mixed bacteria were cultured from coelomic swabs, with Myroides species again the most common (63.5%). Histologic examination was carried out on 292 snails. Autolysis affected almost 90% of those found dead but only 3.4% of euthanized snails. Histology commonly identified microsporidial sporocysts in the digestive gland and midgut epithelium of all but two species. Intracellular, extracytoplasmic Cryptosporidium-like organisms were also common in the midgut but were only observed when snails were fixed in 10% formalin (2017–2019), not ethanol. There were no clear pathologic changes associated with either organism. Pigmented hemocytic nodules were commonly observed, most frequently in the foot process; these were either age related or evidence of prior chronic inflammatory reaction and of low clinical significance. With no evidence of poor health and no significant organisms found, a total of 4,978 individuals representing 12 species were exported for reintroduction.
The Masai giraffe (Giraffa camelopardalis tippelskirchi) is endangered in the wild, and successful reproduction in managed care is important to help maintain assurance populations of this highly charismatic subspecies. Detection of pregnancy in giraffes using hormonal monitoring requires multiple samples and cannot distinguish pregnancy from pseudopregnancy. A novel enzyme-linked immunosorbent assay that can detect pregnancy-specific protein B (PSPB) for pregnancy diagnosis with a single serum sample was developed from a reticulated giraffe (Giraffa camelopardalis reticulata) placenta. Seventy-eight serum samples were analyzed from three female Masai giraffes before and during five gestation periods that resulted in live calf births. Using an optical density cutoff of 0.2, the assay showed a sensitivity of 93% and specificity of 100% for all samples tested. At 59 d of gestation, sensitivity increased to 100%. The earliest pregnancy detection was at 40 d of gestation. This study documents the successful development of a blood-based PSPB assay for pregnancy diagnosis in Masai giraffe, which can help advance conservation efforts in this endangered species.
Renal disease is an important cause of morbidity and mortality in managed black-footed ferrets (BFF; Mustela nigripes).4,6,12 The objectives of this study were to establish reference intervals for blood analytes of clinically normal BFF (1–2 yr old), summarize the frequency of various renal histopathologic findings in a managed population of BFF, assess the diagnostic performance of blood analytes and urine specific gravity (USG) for the diagnosis of renal disease, and assess if comorbidities or age affects the performance of these analytes in diagnosing renal disease. Reference intervals were established using a cohort (n = 35) of clinically normal, young adult BFF. Postmortem records for all BFF at the Phoenix Zoo between 2001 and 2020 were reviewed, and those with available blood analyte data within 2 wk of death were included (n = 89). Ferrets were placed into one of three groups, based on the organ location of histopathologic abnormalities following necropsy: renal disease as the primary change; those with renal disease and at least one other affected major organ system; or absence of abnormalities in the kidneys. In ferrets with substantial renal changes, the primary diagnosis was amyloidosis (29 of 39; 74.4%). Creatinine, blood urea nitrogen, phosphorus (P), calcium (Ca), Ca:P ratio, USG, globulins, and cholesterol were the best-performing analytes for the diagnosis of renal disease, with an area under the curve of at least 0.90 (95% CI $ 0.80, 1.00). Serum renal markers were within reference intervals in BFF that died without histologic evidence of renal disease. Several blood analytes were significantly affected by age in animals that died of renal disease. This study provides reference intervals for blood analytes in young adult clinically normal BFF and illustrates the clinical utility for the diagnosis of renal disease in this species, particularly creatinine, USG, and P.
The Rhynonyssid mesostigmatic mite, Sternostoma tracheacolum, is a well-documented endoparasitic hematophagous arthropod of the respiratory tracts of multiple avian species, particularly Estrildid finches and canaries. In this retrospective study, 175 medical and 278 pathology records for the Gouldian finch (Chloebia gouldiae) population (N = 377) at the San Diego Zoo between 2013 and 2021 were analyzed to evaluate the effectiveness of ivermectin-based prophylaxis. A multivariable negative binomial regression model was constructed to evaluate the population effects of monthly treatments on morbidity or mortality associated with respiratory mites. While controlling for other factors in the model, the prophylactic treatment did not significantly reduce the monthly rate of mite-associated morbidity or mortality (IRR = 1.017, 95% CI: 0.997-1.036, P = 0.0759); however, low proportions of the population were prophylactically treated over time. Different factors were significant when separately evaluating adjusted associations with respiratory morbidity and mortality. The findings suggest increased rates of respiratory morbidity for each successive year of the study period (IRR = 1.180, 95% CI: 1.046–1.342, P = 0.0090) and increased rates of mite-associated mortality occurring annually between May and October (IRR = 1.697, 95% CI: 1.034–2.855, P = 0.0404) compared to the wet winter season. Our findings highlight the need to continually evaluate and optimize treatment regimens in zoological collections. Further investigations into this host-parasite relationship and potential treatments and preventive therapies are warranted.
Ashley Richardson, Liza Dadone, Matthew Johnston, Priya Bapodra-Villaverde, Amy Schilz, Eunice Contreras, Anne Rivas, Sarah Schwenzer, Stephanie Zec, Brenda Cordova, Sara Ferguson, Krista E. Banks, Daniel L. Gustafson, Miranda J. Sadar
Growing resistance to current antiparasitic medications, both in livestock and in zoological species under human care, makes it imperative to evaluate available drugs on the market, such as eprinomectin. In this prospective study, five males and one female of reticulated (Giraffa reticulata; n = 2), Masai (Giraffa tippelskirchii; n = 1), Nubian (Giraffa camelopardalis; n = 2), and hybrid subspecies (n = 1) of giraffe, received 1.5 mg/kg eprinomectin topically along the dorsum. Using high-performance liquid chromatography, concentrations of eprinomectin in plasma samples collected at 0, 4, 24, and 48 h, and 7, 14, 21, and 28 d were evaluated following drug administration. Complete blood cell counts and biochemistry panels were performed before (n = 6) and after (n = 3) eprinomectin administration. Samples for modified double centrifugal fecal flotation (n = 6) were evaluated prior to eprinomectin administration to evaluate for endoparasites and were repeated after the study (n = 5). Noncompartmental pharmacokinetic analysis was applied to the data. The observed maximum plasma concentration was 11.45 ng/ml and the time of observed maximum concentration was 2.67 d. The mean terminal half-life was 5.16 d. No adverse effects were observed related to eprinomectin administration and no blood work changes were observed. Parasite loads decreased (n = 3) or did not change (n = 2) after eprinomectin administration. The mean peak plasma concentration of eprinomectin in giraffe was similar to that achieved in cattle, despite using three times the dose.
Firocoxib is a COX-2–selective nonsteroidal anti-inflammatory drug (NSAID) with limited effects on COX-1, which means it likely has fewer side effects than typically associated with other NSAIDs. This study determined possible doses of firocoxib based on single- and multidose pharmacokinetic trials conducted in 10 Asian elephants (Elephas maximus). Initially, two single oral dose trials (0.01 and 0.1 mg/kg) of a commercially available tablet (n = 6) and paste (n = 4) formulation were used to determine a preferred dose. The 0.1 mg/kg dose was further evaluated via IV single dose (n = 3) and oral multidose trials (tablets n = 6; paste n = 4). Serum peak and trough firocoxib concentrations were also evaluated in Asian elephants (n = 4) that had been being treated for a minimum of 90 consecutive days. Key pharmacokinetic parameters for the 0.1 mg/kg single-dose trials included mean peak serum concentrations of 49 ± 3.3 ng/ml for tablets and 62 ± 14.8 ng/ml for paste, area under the curve (AUC) of 1,332 ± 878 h*mg/ml for tablets and 1,455 ± 634 h*mg/ml for paste, and half-life (T1/2) of 34.3 ± 30.3 h for tablets and 19.9 ± 12.8 h for paste. After 8 d of dosing at 0.1 mg/kg every 24 h, pharmacokinetic parameters stabilized to an AUC of 6,341 ± 3,003 h*mg/ml for tablets and 5,613 ± 2,262 for paste, and T1/2 of 84.4 ± 32.2 h for tablets and 62.9 ± 2.3 h for paste. Serum COX inhibition was evaluated in vitro and ex vivo in untreated elephant plasma, where firocoxib demonstrated preferential inhibition of COX-2. No adverse effects from firocoxib administration were identified in this study. Results suggest administering firocoxib to Asian elephants at a dose of 0.1 mg/kg orally, using either tablet or paste formulations, every 24 h.
The objective of this study was to determine the pharmacokinetics of two orally administered doses of tramadol (1 mg/kg and 5 mg/kg) and its metabolite, O-desmethyltramadol (M1) in giant tortoises (Chelonoidis vandenburghi, Chelonoidis vicina). Eleven giant tortoises (C. vandenburghi, C. vicina) received two randomly assigned, oral doses of tramadol (either 1 mg/kg or 5 mg/kg), with a washout period of 3 wk between each dose. The half-life (t½) of orally administered tramadol at 1 mg/kg and 5 mg/kg was 11.9 ± 4.6 h and 13.2 ± 6.1 h, respectively. After oral administration of tramadol at 1 mg/kg and 5 mg/kg, the maximum concentration (Cmax) was 125 ± 69 ng/ml and 518 ± 411 ng/ml, respectively. There were not enough data points to determine pharmacokinetic (PK) parameters for the M1 metabolite from either dose. Tramadol administered orally to giant tortoises at both doses provided measurable plasma concentrations of tramadol for approximately 48 h with occasional transient sedation. Oral tramadol at 5 mg/kg, on average, achieves concentrations of >100 ng/ml, the reported human therapeutic threshold, for 24 h. Based on the low levels of M1 seen in this study, M1 may not be a major metabolite in this taxon.
Hematology is a routine component of clinical management in veterinary patients. Anticoagulant choice can profoundly influence morphologic assessment of erythrocytes, leukocytes, thrombocytes, and their subsequent quantification. Previous chelonian studies suggest that lithium heparin (LH) is a superior anticoagulant due to hemolysis resulting from dipotassium ethylenediaminetetraacetic acid (dEDTA) in some species. The aim of this study was to compare the effects of dEDTA and LH on hematologic values in Blanding's turtles (Emydoidea blandingii, n = 35), painted turtles (Chrysemys picta, n = 34), and common snapping turtles (Chelydra serpentina, n = 36). We collected samples from free-ranging turtles and immediately divided whole blood into LH and dEDTA tubes. Packed cell volume, total solids, erythrocyte sedimentation rate, white blood cell counts, and differential leukocyte counts were determined. Hemolysis was observed macro- and microscopically in dEDTA samples from painted turtles and common snapping turtles. Packed cell volume and heterophil:lymphocyte was lower and erythrocyte sedimentation rate was higher in LH samples from painted turtles (p, 0.05). In snapping turtles, the PCV, number of monocytes, and number of eosinophils was lower in LH samples (p, 0.05). In Blanding's turtles, the number of eosinophils and basophils was higher in LH samples, while heterophil counts were lower (p, 0.05). Anticoagulant choice created constant and proportional bias for multiple analytes in a species-dependent fashion. LH is the recommended anticoagulant for hematology in painted turtles and common snapping turtles. Either LH or dEDTA may be used in Blanding's turtles, though anticoagulant-specific reference intervals may be necessary.
The Houston toad (Anaxyrus houstonensis), a primarily terrestrial amphibian of south-central Texas, has been listed as federally endangered since 1970. Sedation is an important tool for obtaining diagnostics and providing treatment in this species. This prospective, randomized, and blinded study compared the sedative effects of SC alfaxalone (Protocol A) at approximately 12 mg/kg (median [range] = 12.70 [12.09–13.95] mg/kg] to SC alfaxalone–dexmedetomidine (Protocol AD) at approximately 12 mg/kg (median [range] = 12.68 [12.16–13.56] mg/kg) and 0.1 mg/kg (median [range] = 0.1 [0.07–0.13] mg/kg), respectively, in adult Houston toads (n = 26). Toads from Protocol AD received atipamezole SC at approximately 1 mg/kg (median [range] = 0.96 [0.75–1.25] mg/kg) 45 min postinduction, whereas toads from Protocol A received the equivalent volume of SC sterile saline at the same time point. Heart rate, gular rate, and times to first effect, loss of righting reflex, ability to position for radiographs, loss of nociception, return of righting reflex, and full recovery were recorded. A significantly greater number of toads lost righting reflex, positioned for radiographs, and lost nociception with Protocol AD compared with Protocol A. Additionally, time to return of righting reflex and time to full recovery were significantly longer with Protocol AD than with Protocol A. The protocols did not differ significantly in time to first effect, time to radiographic positioning, or time to loss of nociception. Histologic examination of four toads euthanized during the study revealed acute injection site reactions from all administered drugs, including saline. No clinical adverse reactions were observed. This study demonstrates that the combination of SC alfaxalone and dexmedetomidine results in deeper sedation than SC alfaxalone alone, but also correlates with longer recovery times despite antagonist administration.
This randomized, crossover study evaluated three sedation protocols administered subcutaneously in nine budgerigars (Melopsittacus undulatus) and nine black-cheeked lovebirds (Agapornis nigrigenis). All protocols included midazolam (5 mg/kg), combined with butorphanol (5 mg/kg) (BM), medetomidine (20 lg/kg) (MM), or alfaxalone (13 mg/kg) (AM). Mortalities from suspected cardiorespiratory arrest were observed when AM was used in lovebirds, even after reduction of alfaxalone dosage to 3 mg/kg, and therefore this protocol was excluded from further use in this species. Induction and recovery times were recorded and their quality assessed. Sedation depth and heart and respiratory rates were measured every 5 min and radiographic positioning was attempted at 10 and 20 min. At 30 min, midazolam and medetomidine were reversed with flumazenil (0.05 mg/kg, SC), and atipamezole (0.2 mg/kg, SC), respectively. MM consistently provided deep sedation in both species, with successful radiographic positioning at every attempt. As expected, heart rate was often lower with MM than with other protocols, but no associated complications were noted. In budgerigars, BM had the lowest radiographic positioning success rate (10 min: 5/9, 20 min: 3/9), whereas in lovebirds it provided significantly deeper sedation (P < 0.001), allowing radiographic positioning in all subjects. In both species, BM provided the shortest recovery times. AM resulted in reliable radiographic positioning of all budgerigars at 10 min, but not at 20 min (5/ 9), and provided consistently poor recoveries. This study highlights how differently two psittacine species of similar size may react to the same sedation protocols. AM sedation cannot be fully reversed and produced significant undesirable effects, several of which have been previously reported with alfaxalone administration to avian species. The authors therefore caution against using alfaxalone–midazolam combinations in budgerigars and black-cheeked lovebirds. Both BM and MM provided reliable sedation in these species, and appear to be suitable alternatives to AM.
Salene A. Colombo, Daniela M.D. de Mello, Bruna R.M. Morais, Lauranne A. Salvato, Fernanda A. Dorella, Guilherme C. Tavares, Vera M.F. da Silva, Maria I. de Azevedo
The present study characterized the filamentous and yeast-like fungal microbiota of the nasal cavity and rectum of Amazonian manatees (Trichechus inunguis) undergoing rehabilitation at the Laboratory of Aquatic Mammals, National Institute of Amazonian Research, Manaus, Amazonas, and determined the antifungal susceptibility of these organisms. Nasal and rectal swabs were collected from 22 calves and three juveniles. The samples were seeded in Sabouraud agar supplemented with chloramphenicol 10%, incubated at 26°C, and observed daily for up to 7 d. The growth of different filamentous and yeast-like fungi was observed among the two anatomical sites. Filamentous fungi were categorized by macro- and microscopic characteristics of the colonies. Representatives of each group were selected for molecular identification based on the internal transcribed spacer region. Yeast identification was performed using MALDI-TOF MS and molecular analyses. Thirteen genera of filamentous fungi and six genera of yeasts were isolated and identified. The dominant filamentous species were Fusarium spp., Aspergillus spp., and Cochliobolus lunatus in the nostril samples and Aspergillus melleus in the rectal samples. Candida was the dominant genus among the identified yeasts at both anatomical sites. In the antifungal susceptibility test, 28 isolates showed resistance to fluconazole (78%), itraconazole (39%), and nystatin (42%). The knowledge of fungal microbiota composition of Amazonian manatees provides information that assists in monitoring the health status of individuals maintained in captivity, as these organisms can behave either as opportunists or as primary pathogens. Moreover, the composition and resistance of these organisms may vary among different rehabilitation institutions or different time frames of search, reinforcing the importance of constant in loco surveillance of these microorganisms. This study provides new perspectives on the fungal diversity in the microbiota of manatees and supports future studies concerning the clinical and epidemiological aspects and the impacts of these agents on the health of Amazonian manatees undergoing rehabilitation.
A mixture of butorphanol, azaperone, and medetomidine (BAM) is frequently used for immobilization of North American hoofstock. Common adverse effects include respiratory depression, hypoxemia, and bradycardia. In this nonblinded crossover study the efficacy of two a-2 adrenergic antagonists, tolazoline and vatinoxan, were evaluated in alleviating adverse effects of BAM in Rocky Mountain elk (Cervus canadensis). Early administration of these antagonists was hypothesized to cause an increase in heart rate, respiratory rate, partial pressure of oxygen (PaO2) and hemoglobin oxygen saturation (SpO2), as well as reduction in mean arterial blood pressure without affecting sedation levels. Eight captive adult female elk were immobilized on three separate occasions at least 14 d apart with 0.15 mg/kg butorphanol, 0.05 mg/kg azaperone, and 0.06 mg/kg medetomidine. Tolazoline (2 mg/kg IM), vatinoxan (3 mg/mg medetomidine IV) or sterile saline (2 ml IM) were administered 20 min postinduction. The BAM caused hypoxemia, bradycardia, and moderate hypertension, and because of the severe hypoxemia observed, all animals received intratracheal oxygen throughout immobilization. Heart rate, respiratory rate, rectal temperature, SpO2, PaO2, and systolic, diastolic, and mean arterial blood pressure were monitored every 5 min throughout the immobilization. Intramuscular tolazoline caused a brief but significant drop in mean arterial pressure compared with controls and a brief but nonsignificant increase in heart rate. Vatinoxan caused a significant drop in blood pressure and a brief significant increase in heart rate. Changes in respiratory rates and PaO2 were not observed with either antagonist; however, all animals received oxygen, which may have influenced this result. The depth of sedation was unchanged after administration of either drug.
Based upon previous clinical experience with domestic cats (Felis catus), the ability to assess ABC blood types and blood (in-)compatibilities of nondomestic felids, and adequately consider and plan for blood transfusions, may be important. Although nondomestic felids appear to have an ABC blood group system similar to domestic cats, typing with point-of-care kits and by CMAH genotyping for domestic cats have not been reported. In this study, 162 blood samples from 18 different nondomestic felid species (cheetah [Acinonyx jubatus, n = 42], lion [Panthera leo, n = 33], tiger [Panthera tigris, n = 23], Canada lynx [Lynx canadensis, n = 11], snow leopard [Uncia uncia, n = 10], puma [Puma concolor, n = 7], clouded leopard [Neofelis nebulosa, n = 6], serval [Leptailurus serval, n = 5], jaguar [Panthera onca, n = 5], fishing cat [Prionailurus viverrinus, n = 4], Pallas cat [Felis manul, n = 3], bobcat [Lynx rufus, n = 3], ocelot [Leopardus pardalis, n = 3], black footed cat [Felis nigripes, n = 2], leopard [Panthera pardus, n = 2], African wildcat [Felis lybica, n = 1], caracal [Caracal caracal, n = 1], and sand cat [Felis margarita, n = 1]) were ABC blood typed by laboratory and point-of-care tests, genotyped for four known CMAH variants for type B and type C (AB) phenotypes, and crossmatched with one another and domestic type A cats. Traditional tube typing identified blood type A (n = 106), type B (n = 8), type C (n = 43), and no discernible ABC type (n = 4). Several discrepancies were found between point-of-care and traditional typing test results. None of the tested felids possessed the four CMAH variants responsible for type B and C (AB) in domestic cats. Crossmatch incompatibilities (≥2+ agglutination) were identified within and between nondomestic felid species and beyond ABC incompatibilities. Of 26 crossmatches performed between domestic cats and various nondomestic felids, only 7 (27%) were compatible. In conclusion, point-of-care typing kits and CMAH genotyping, successfully used in domestic cats, may not identify the correct ABC blood type in nondomestic felids. Prior crossmatching is recommended to increase the likelihood of compatible transfusions between any nondomestic felids.
Meerkats (Suricata suricatta) housed at two accredited zoological institutions in the United States were evaluated via echocardiography, thoracic radiography, and blood biomarkers—taurine and feline N-terminal pro-B-type natriuretic peptide—to determine the prevalence and severity of dilated cardiomyopathy (DCM) in both populations. In total, 24 meerkats were evaluated and 7 were diagnosed with DCM based on the following parameters: left ventricular internal diameter at end diastole > 1.30 cm, left ventricular internal diameter at end systole > 1.10 cm, and a fractional shortening of <18%. Echocardiographic parameters were identified and reported for normal and affected meerkats, whereas thoracic radiographs were not useful for screening for DCM. Meerkats with DCM were treated with pimobendan and/or benazepril and furosemide if indicated. Seven meerkats died during the study period, with the majority exhibiting myocardial fibrosis. Of the blood parameters tested, elevated taurine levels were associated with DCM. Further research is necessary to characterize the etiology of DCM in meerkats.
Elephant endotheliotropic herpesvirus (EEHV) can induce fatal hemorrhagic disease (HD) in African elephants (Loxodonta africana). Once clinical signs develop, progression is rapid, even with aggressive treatment. There is a critical need to develop point-of-care diagnostic tests to aid in identification of EEHV-HD prior to the onset of overt clinical signs. Study objectives were to investigate a novel, point-of-care viscoelastic coagulation monitor (VCM Vet), compare the results to thromboelastography (TEG), and report traditional hemostatic analytes in adult African elephants. Whole blood was collected from seven clinically healthy elephants (four females and three males, 18–47 yr) and analyzed in duplicate via VCM Vet and kaolin-activated TEG 1–3 and 30 min following collection, respectively. Separated plasma was frozen for ancillary coagulation testing. Both analyses generated quantifiable clotting reactions with variables (median [range]) describing clot formation rate (VCM Vet, clot time = 682 s [530–987 s], clot formation time = 244 s [186–744 s], Alpha = 40° [14–47°]; TEG, reaction time = 6.2 min [3.7–11.8 min], kinetic time = 1.3 min [0.9–2.6 min], Alpha = 70° [57–77°]), clot strength (VCM Vet, maximum clot formation = 34 units [20–45 units]; TEG, maximum amplitude = 75 mm [69–80 mm], shear elastic modulus strength = 14.7 Kdynes/s [11.3–19.5 Kdynes/s]), and clot lysis (VCM Vet, lysis index at 30 min = 100% [100–99%], lysis index at 45 min = 98% [95–100%]; TEG, lysis index at 30 min = 0% [0–0.4%], lysis index at 60 min = 1.4% [0–2.6%]) recorded. Additional testing (median [range]) included D-dimer concentration (33 ng/ml [28–94 ng/ml]), prothrombin time (12.4 s [12.2–13.2 s]), activated partial thromboplastin time (17.2 s [14.2–18.8 s]), and fibrinogen concentration (297 [282–383] mg/dL). Tracings generated by VCM Vet and TEG were clinically similar, and there was visual agreement and minimal difference between quantitative variables for duplicate tests. VCM Vet is a promising, user-friendly tool for use in identification and management of coagulopathies in African elephants.
Detailed knowledge of biological variation can facilitate accurate interpretation of clinical pathology parameters. A recent biological variation study in Asian elephants (Elephas maximus) found that hematology parameters had high individuality, which suggests that population-derived reference intervals may be an insensitive diagnostic tool. In elephant medicine, sensitive hematology-related diagnostics are crucial for clinical decision-making, particularly in elephants at risk for elephant endotheliotropic herpesvirus hemorrhagic disease (EEHV-HD). The objective of this study was to assess biological variation of hematology parameters in African elephants to determine whether population-derived reference intervals are a sensitive diagnostic tool for interpreting results and to provide a useful alternative. Eight healthy African elephants had blood collected under behavioral training every other week for 8 wk. Complete blood cell count (CBC) analysis was performed in duplicate to assess analytical variation. Previous methods were used to determine between-individual variation, within-individual variation, index of individuality, and reference change values (RCV). This study found that most hematology parameters displayed intermediate-to-high individuality, which suggests that alternatives to population-derived reference intervals are necessary to detect pathologic changes. To test the results of our biological variation data, a case of EEHV-HD was retrospectively evaluated. Individual normal values and calculated RCV detected a clinically significant monocytopenia, leukopenia, and thrombocytopenia associated with EEHV2 viremia. However, none of these parameters fell outside a population-derived reference interval. This study highlights the utility of biological variation in clinical decision-making and demonstrates that individual normal values and RCV may be important diagnostic tools for CBC interpretation in African elephants.
This study examined the viral shedding kinetics of elephant endotheliotropic herpesvirus (EEHV) in African elephants (Loxodonta africana) compared to viral shedding behavior in Asian elephants (Elephas maximus). Little is known about the transmission dynamics and epidemiology of this disease in African elephants. In light of recent clinical cases and mortalities, this paper aims to identify trends in viral biology. Trunk wash samples were collected from 22 African elephants from four North American zoological institutions that had recently experienced herd viremias or translocations. Processing of these samples included DNA extraction followed by qPCR to quantitate viral DNA load. The results were then compared with available literature that chronicled similar cases in Asian and African elephants. Minimal EEHV shedding was detected in response to varied herd translocations. Increased shedding was recorded in herds in which an elephant experienced an EEHV viremia when compared to baseline shedding. These index infections were followed by subsequent viremias in other elephants, although it is not known if these were recrudescence, transient controlled viremias, and/or primary infections via transmission to other elephants. When compared to historically published data, it was observed that EEHV3 cases in African elephants and EEHV1A cases in Asian elephants had consistently higher levels of viral DNA in the blood than were shed in trunk secretions, a fact that is seemingly inconsistent with such severe cases of disease and the high mortality rates associated with those respective types. The findings produced in this study highlight the need for more routine monitoring of viral shedding in African elephant herds to elucidate possible EEHV transmission and recrudescence factors for ex situ population management.
Heart disease in birds contributes to premature death and is usually detected postmortem. Echocardiography is a sensitive and noninvasive diagnostic modality but reported standard values for many species of birds, including American flamingos (Phoenicopterus ruber), are unavailable. Echocardiography was opportunistically performed on 30 unsedated American flamingos during their annual routine health examination. Structural heart disease was not found in any of the examined birds. However, 18 birds showed varying degrees of dynamic intraventricular obstruction. Echocardiographic parameters are reported. Benign neurocardiogenic weakness or syncopal events during handling were suspected in three birds. Stress combined with an intraventricular obstruction is believed to have triggered a cascade of parasympathetic innervation and sympathetic inhibition, similar to neurally mediated reflex syncope in humans.
The maned sloth (Bradypus torquatus) is an endemic and endangered species of two Brazilian states, with much unknown biological information needed to direct conservation actions. Other sloth species have been studied regarding anesthesia; however, there is a lack of anesthesia research for the maned sloth. Anesthetic data were collected from 12 free-range maned sloths that were immobilized for a field examination. Individuals were anesthetized using a combination of ketamine (4.0 mg/kg) and medetomidine (0.03 mg/kg), and antagonized with atipamezole (0.1 mg/kg). Time to induction and recovery were recorded and compared with sex and age classes. After the induction and until antagonist administration, physiological parameters (rectal temperature, heart rate, respiratory rate, and oxygen saturation) were recorded every 10 min during anesthesia and were statistically evaluated over time. Induction was fast (3.21 ± 0.76), but recovery was longer (113.3 ± 18) when compared to other studies. Induction and recovery times were not different across sex or age classes. Rectal temperature, heart rate, and oxygen saturation remained stable throughout the procedure. Respiratory rate significantly decreased over time, from 18.25 ± 7.03 to 13.17 ± 3.66 movements per minute. Our results indicate that the described combination of ketamine and medetomidine is a safe and effective choice for anesthesia of maned sloths.
Sedation, recovery response, and physiologic outcomes were evaluated in five captive reindeer (Rangifer tarandus) in Minnesota using a completely reversible immobilization protocol. Reindeer were immobilized with butorphanol (0.23–0.32 mg/kg), midazolam (0.23–0.32 mg/kg), and medetomidine (0.15 mg/kg) (BMM) via IM dart. Induction time (IT), recumbency time (DT), and recovery time (RT) were recorded. Temperature (T), respiratory rate (RR), pulse rate (PR), pulse oximetry (SpO2), arterial blood gas values including oxygen (PaO2), and carbon dioxide (PaCO2) tensions and lactate (Lac) were recorded preoxygen supplementation and 15 min postoxygen supplementation. Reversal was done using naltrexone (2.3–3.0 mg/kg), flumazenil (0.008–0.01 mg/kg) and atipamezole (0.62–0.78 mg/kg) (NFA) IM, limiting recumbency to 1 h. Median IT, DT, and RT were 5 min, 46 min, and 7 min, respectively. SpO2 (92 to 99%, P = 0.125), PaO2 (45.5 to 97 mmHg, P = 0.25), and PaCO2 (46.5 to 54.6 mmHg, P = 0.25) all increased, whereas Lac (3.02 to 1.93 mmol/L, P = 0.25) decreased between baseline and 15 min postoxygen supplementation, without statistical significance. BMM immobilization, and reversal with NFA provided rapid and effective immobilization and recovery, respectively. Oxygen supplementation mitigated hypoxemia in all reindeer.
Improved methods are needed to prevent wildlife deaths from anthrax. Caused by Bacillus anthracis, naturally occurring outbreaks of anthrax are frequent but unpredictable. The commercially available veterinary vaccine is labeled for subcutaneous injection and is impractical for large-scale wildlife vaccination programs; therefore, oral vaccination is the most realistic method to control and prevent these outbreaks. We reported the induction of an anthrax-specific lethal toxin (LeTx) neutralizing antibody response in mice following oral vaccination with alginate microcapsules containing B. anthracis Sterne strain 34F2 spores, coated with poly-L-lysine (PLL) and vitelline protein B (VpB). We continued evaluating our novel vaccine formulation through this proof-of-concept study in white-tailed deer (WTD; Odocoileus virginianus; n = 9). We orally vaccinated WTD via needle-free syringe with three formulations of the encapsulated vaccine: 1) PLL-VpB–coated microcapsules with 107–8 spores/ml (n = 5), 2) PLL-VpB–coated microcapsules with 109–10 spores/ml (n = 2), and 3) PLL-coated microcapsules with 109–10 spores/ml (n = 2). Although the limited sample sizes require continued experimentation, we observed an anthrax-specific antibody response in WTD serum following oral vaccination with PLL-coated microcapsules containing 109 spores/ ml. Furthermore, this antibody response neutralized anthrax LeTx in vitro, suggesting that continued development of this vaccine may allow for realistic wildlife anthrax vaccination programs.
With vulture population numbers on the decline globally, many countries resort to supplementary feeding to maintain colony health. Despite what is perceived as adequate feeding in South Africa, colonies are still characterized by poor breeding success. One reason could be that supplementary sites fail to meet micronutrient needs of birds. With results from zoological gardens indicating that some carcasses are low in their vitamin E concentrations, vitamin deficiencies may be an underlying problem. For this study it was determined if the feeding of whole pig carcasses, a common food item, could have a negative effect on plasma vitamin E concentrations in a captive colony. Plasma vitamin E concentrations were 7.38 ± 2.92 and 4.51 ± 1.24 after feeding whole pig carcasses (n = 14). Behaviorally, the birds also avoided the viscera and fat when feeding. Reasons for their low vitamin E concentrations could have resulted from the birds consuming only the pork meat, which is known to be low in vitamin E, or from natural peroxidation because of the high fat content of the carcasses. The study thus highlights the need for further research to ascertain the impact of feeding pig carcasses on wild vultures feeding routinely at supplementary feeding sites and also for considerations towards vitamin E supplementation.
Ten cases of small intestinal tympany with or without volvulus were documented in Central American river turtles (Dermatemys mawii). These turtles were under managed care at the Philadelphia Zoo in Philadelphia, PA, USA, with one case followed after transfer to a different institution. The primary clinical presentation was abnormal buoyancy in nearly all cases (9/10) and anorexia in two cases (2/10). Five of 10 turtles with suspected or radiographically confirmed small intestinal tympany recovered (5/10), whereas five cases (5/ 10) resulted in death or euthanasia. In all fatal cases (5/5), small intestinal volvulus was identified at gross necropsy, with concurrent colonic volvulus identified in 2/5 cases. Other notable necropsy findings were hepatic lipidosis (3/5) and thrombosis of intestinal or renal vasculature (2/5). In all fatal cases (5/5), there was short (1 to 2 days) clinical progression from abnormal buoyancy to death or euthanasia. In the majority of cases (6/10), an abrupt change in diet, notably the overfeeding of fresh fruit or excessive amounts of mulberry (Morus spp.) browse, or ingestion of indigestible foreign material, occurred prior to presentation. Temporary suboptimal environmental temperatures were suspected prior to the onset of clinical signs in 4/10 cases. Optimal husbandry conditions including nutrition and environmental temperature appear vital to preventing this condition. Recognition of early clinical signs of this condition, such as abnormal buoyancy and anorexia, and environmental correction or medical therapy, may prevent fatality and result in a better outcome in these cases.
This case series investigates a cluster of deaths in a captive colony of Leschenault's rousettes (Rousettus leschenaultii). Six of seven bats that died between March and September 2021 were diagnosed postmortem with both iron overload (IO) and neoplasia, neither of which have previously been reported in this species. Iron status was assessed via hepatic histopathological grading, hepatic iron concentration, and, in two cases, serum iron concentration. On histopathological grading, all cases had hemochromatosis except one, which had hemosiderosis. Hepatic iron concentrations did not correlate with histopathological grading. Neoplasms in these six bats included hepatocellular carcinoma (HCC; 4), bronchioloalveolar adenocarcinoma (1), pancreatic adenocarcinoma (1), and sarcoma of the spleen and stomach (1). One bat had two neoplasms (HCC and sarcoma of the spleen and stomach). One additional case of HCC in 2018 was identified on retrospective case review. Etiology was investigated to the extent possible in a clinical setting. Nutritional analysis and drinking water testing found oral iron intake within acceptable bounds; however, dietary vitamin C was potentially excessive and may have contributed to IO. Panhepadnavirus PCR testing of liver tissue was negative for all bats. A species-associated susceptibility to IO, as seen in Egyptian fruit bats (Rousettus aegyptiacus), is possible. The high incidence of HCC is suspected to be related to IO; other differentials include viral infection. Causes or contributing factors were not definitively identified for the other neoplasms seen but could include age, inherited risk (given a high level of inbreeding), or an oncogenic virus. Pending further research in this species, it is recommended that keepers of Leschenault's rousettes offer conservative amounts of vitamin C and iron (as for Egyptian fruit bats), submit for postmortem examination any euthanized or found dead, and share records of similar cases.
The striped bass (Morone saxatilis) has been a fish species of special concern in Canada since its marked decline in the early 21st century in the St. Lawrence River. Individuals kept in public aquaria contribute to public education and could support conservation efforts through research. Over a 3-yr period, 12 male striped bass housed in a multispecies exhibit developed coelomic distension. The testes were enlarged (12/12), cystic (2/12), and heterogeneous (3/12) on coelomic ultrasound. Upon coeliotomy, enlarged (12/12), partially (4/12) or totally white discolored (6/12) testes were noted. These were associated with coelomic hemorrhage (8/12), effusion (3/12) or adhesions to surrounding organs (9/12). Orchiectomies were performed in all fish. Among these, seven fish survived 2 mon postsurgery, and four fish were still alive 900 d postsurgery. Germ cell neoplasia was diagnosed on histopathological examination in 9 of 12 individuals, but no abnormalities were found in the three other cases. Preventive orchiectomies were performed on the remaining six male striped bass in this exhibit. Germ cell neoplasms were present in two of these six fish. No anesthetic or surgical complications were noted; all six cases were alive 2 mon postsurgery and four of the fish survived 900 d postsurgery. Survival times were not significantly different between fish that underwent preventive or curative orchiectomy (P = 0.19). Although risk factors associated with the development of these gonadal tumors remain unknown, a genetic or environmental origin is suspected. Orchiectomy should be considered in suspected cases of testicular tumors.
Paula Rodriguez, Danielle K. Tarbert, Frank Ridgley, Kenneth J. Conley, James Steeil, Rachel E. Burns, Meg Sutherland-Smith, Karen Rosenthal, Joanne R. Paul-Murphy
Four green iguanas (Iguana iguana) and one blue iguana (Cyclura lewisi) from five facilities were diagnosed with sodium urate cholelithiasis. One case was diagnosed antemortem via ultrasonography, and the iguana underwent a choledochotomy for treatment. The other four cases were identified at necropsy. Pathologic hepatic and biliary changes were present in four of the five cases at necropsy. Histologically, four iguanas had hepatic fibrosis, three had bile duct hyperplasia, and one had cholangiohepatitis and pancreaticocholedochitis. Two iguanas had pathologic renal changes. This is the first report of sodium urate cholelithiasis in reptiles. This case series highlights the potential significant clinical disease caused by sodium urate cholelithiasis and the importance of biliary system evaluation. Further investigation is recommended to explore the pathogenesis of reptilian sodium urate cholelith formation.
Listeria monocytogenes is an ubiquitous environmental saprophytic bacterium causing listeriosis in domestic animals, humans, and occasionally wildlife. In animals, this foodborne zoonotic disease mainly occurs in ruminants and it is rare in carnivores. Seven red foxes (Vulpes vulpes) and one Eurasian lynx (Lynx lynx) were diagnosed with listeriosis between 2010 and 2021 at the Institute for Fish and Wildlife Health, Bern, Switzerland. Necropsy and histopathology revealed meningitis (six of seven red foxes), hepatitis (six of seven red foxes), pneumonia (five of seven red foxes), splenitis (two of seven red foxes) and splenomegaly (the Eurasian lynx, two of seven red foxes). Listeria monocytogenes was isolated from either lung, spleen, liver, or kidney of all animals. Serotyping detected L. monocytogenes serotype 1/2a in five red foxes and the Eurasian lynx and serotype 4b in two red foxes. Six red foxes were positive for canine distemper virus (CDV) by polymerase chain reaction, whereas the Eurasian lynx and one red fox were negative. One red fox that was positive for CDV and listeriosis was also diagnosed with salmonellosis. The identified L. monocytogenes serotypes are among the three most frequently isolated serotypes (1/2a, 1/2b, and 4b) from food or the food production environment and those that cause most listeriosis cases in humans and animals. Coinfection with CDV in six red foxes questions the role of CDV as potential predisposing factor for septicemic listeriosis. The detection of listeriosis in the regionally endangered Eurasian lynx and in carnivores highly abundant in urban settings, such as red foxes, reinforces the importance of wildlife health surveillance in a One Health context and adds the Eurasian lynx to the list of carnivores susceptible to the disease. Further investigations are required to assess the prevalence and epidemiology of L. monocytogenes in free-ranging carnivores and its interaction with CDV.
Two female (FL 1, FL 2) and one male (ML) 11-wk-old, intact, captive African lion cubs (Panthera leo leo) were presented with a history of mild vestibular signs. Initial serum vitamin A concentrations were low (140 nmol/L) for ML. Calvarial hyperostosis was confirmed using computed tomography (CT) of the head and cervical vertebrae in each cub. CT measurements were adapted in relation to the skull width. ML showed the most pronounced thickening of the tentorium cerebelli and occipital bone, represented by a tentorium cerebelli to skull width ratio (TCR) of 0.08 (FL 1: 0.06, FL 2: 0.05) and a basisphenoid to skull width ratio (BBR) of 0.07 (FL 1: 0.06, FL 2: 0.04). Magnetic resonance imaging (MRI) revealed cerebellar herniation and cervical intramedullary T2-weighted hyperintensity from C1, extending caudally for at least two cervical vertebrae in all cubs. Treatment was initiated with subcutaneous vitamin A supplementation and feeding of whole carcasses. Improvement in ataxia was noticed 3 wk later. Follow-up CT and MRI examinations were performed in ML after 3 and 8 mon. The affected bones appeared slightly less thickened and TCR and BBR had decreased to 0.05 after 3 mon. The cerebellum remained mildly herniated, accompanied by amelioration of cervical T2w hyperintensities. After 8 mon, evaluation and diagnostic imaging revealed further improvement regarding the neurologic status and measurements (TCR 0.05, BBR 0.04) despite persistence of a subtle cerebellar herniation. In conclusion, bone remodeling and improvement in clinical signs may be achievable in young lion cubs presented with calvarial hyperostosis and may be attributable to high-dose vitamin A supplementation.
Joint luxations commonly occur in animals secondary to traumatic injury. Because of the unique hind-limb anatomy of macropods, surgical stabilization of orthopedic injuries is considered challenging, and reports of successful management are limited. A 4-yr-old male neutered red kangaroo (Osphranter rufus) presented with a dorsolateral luxation of the left tibiotarsal joint. Surgical reduction and tarsal arthrodesis were performed. Although the full range of motion of the tarsal joint was limited, this kangaroo was still able to ambulate normally at slow speeds following surgery and recovery. The aim of this report was to describe the surgical and postoperative management of a tibiotarsal luxation in a kangaroo. There were significant postoperative complications in this kangaroo, and antibiotic regional limb perfusion was used to treat wound and implant infection.
Multiple species of elephant endotheliotropic herpesvirus (EEHV) have caused fatal hemorrhagic disease in African (Loxodonta africana) and Asian (Elephas maximus) elephants. To date, EEHV7 has been detected only in benign pulmonary and skin nodules and in saliva of African elephants and has not been associated with clinical illness. Low-level viremia due to EEHV7A was detected via qPCR in two subadult African elephants during routine surveillance. Hematologic changes were noted in both elephants, including leukopenia, lymphopenia, monocytopenia, and band heterophilia. Treatment was initiated with famciclovir, antimicrobials, and rectal fluids, and one elephant received plasma transfusions due to a progressive decrease in platelet count. Both elephants remained asymptomatic throughout the viremias, with rapid resolution of hematologic abnormalities. These cases add to the current understanding of the epidemiology of EEHV in African elephants; to the authors' knowledge, they represent the first documentation of clinical disease due to EEHV7 infection in any elephant.
Two zoo-maintained short-beaked echidnas (Tachyglossus aculeatus) had long histories of intermittent anorexia and lethargy. Case 1 presented with a recurrence of these signs after transfer to another facility and died shortly after arrival. A focal area of hyperattenuation within the paratracheal tissue of the cranial mediastinum was noted antemortem on CT. Postmortem, this corresponded with severe thyroid follicular hyperplasia with lymphoplasmacytic thyroiditis. Additional findings included a systemic fungal infection without an inflammatory response, suggesting underlying factors such as torpor or immunosuppression. In Case 2, an intrathoracic mass was identified during a preshipment examination. CT confirmed a contrast-enhanced mass compressing the cranial vena cava and right atrium, and the animal was euthanized. The mass was diagnosed histologically as thyroid adenocarcinoma. These cases report thyroiditis and thyroid adenocarcinoma in echidna and describe the use of IV contrast and CT as a diagnostic aid in this species.
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