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Obstructive and incidental urolithiasis cases were evaluated in 5 Asian colobine monkey species (n= 21 individuals) that included 12 silvery langurs (Trachypithecus cristatus), 6 spectacled langurs (Trachypithecus obscurus), 1 Javan langur (Trachypithecus auratus auratus), 1 François' langur (Trachypithecus francoisi), and 1 red-shanked douc langur (Pygathrix nemaeus) from eight zoologic institutions. All institutions that responded were Association of Zoos and Aquariums, European Association of Zoos and Aquaria, or World Association of Zoos and Aquariums accredited. Males were more commonly represented in the total number of cases (86%), and all cases of obstructive urolithiasis occurred in males. The most common clinical signs observed in obstructive cases included stranguria (58.8%), lethargy (41.2%), anorexia (29.4%), depression (17.6%), and penile manipulation (11.8%). Clinicopathologic abnormalities revealed azotemia (76.5%), anemia (35.3%), and hyperkalemia (23.5%). Eleven of the 21 cases included urinalysis results, and crystalluria was reported in all 11. Obstructive cases were more commonly managed surgically, with medical management following. Instances of individual obstruction ranged from 0 (incidental finding) to 18, with a median of 8 (mean of 4.3) recurrent obstructions. A total of 39 urolith analyses were available from 17 of 21 cases, with calcium carbonate being the most common type isolated (37 of 39, 94.9%). Calcium oxalate was observed in the remaining 5.1% of cases (2 of 39). No cases exhibited a consistent match of crystalluria and urolith type. Death or euthanasia secondary to obstructive urolithiasis occurred in 52.4% of cases. Urinary obstruction secondary to urolithiasis appears to be a relevant cause of morbidity and mortality in Asian colobine species, and further study into etiology and preventive medicine should be undertaken.
Tufted puffins (Fratercula cirrhata) are commonly exhibited in zoologic institutions across the world, yet there is a paucity of information on causes of mortality in managed populations. This retrospective review reports the pathologic findings associated with 91 tufted puffins at a single institution over 35 years from 1982 to 2017. Common pathologic findings were evaluated by age at death, sex, year, and season. With the exception of neonates, the leading pathologic finding across all age classes was aspergillosis, particularly in adults. Hemoparasitism, predation, and trauma were also frequent causes of mortality. Neonatal mortality was common and primarily caused by omphalitis, yolk sac disease, and bacterial septicemia, with most cultures revealing Escherichia coli. This study also provides documentation of mortality in tufted puffins secondary to avian pox and suspected toxoplasmosis. Understanding morbidity and mortality trends within a population allows institutions to form management plans and implement practices to improve outcomes and survival.
Sara Goñi Martinez, Simon Spiro, Amanda Guthrie, Alberto Rodriguez Barbon, Javier Lopez, Mark F. Stidworthy, Daniela Denk, Hannah Jenkins, Taina Strike
The mountain chicken frog (Leptodactylus fallax) is a critically endangered species highly reliant on ex situ conservation programs. In captivity, cholelithiasis has been reported during clinical examinations using diagnostic imaging and observed during postmortem examinations. Some individuals have presented with nonspecific clinical signs potentially associated with this condition, but little is known about its clinical relevance. A retrospective evaluation of ultrasonographic (n = 139) and radiographic (n = 156) images and histopathology samples (n = 32) obtained between 2014 and 2020 from 133 individuals was performed. An ultrasound scoring system was developed to evaluate changes in the gallbladder, with 38% (53/139) of the images showing abnormal contents. Gallbladder distension appeared associated with these findings (generalized estimating equation model, P < 0.001). In contrast, radiography proved to have low overall sensitivity (48%) when compared to ultrasonography for identifying abnormal gallbladder contents. A weak level of agreement (Cohen's kappa = 0.535) was observed between the imaging modalities. Histologically, cholecystitis was present in 86% (12/14) of the individuals, thickening of the gallbladder wall in 71% (10/14), and intrahepatic cholangitis in 90% (18/20). There was no evidence of cholestasis in any of the affected individuals. A significant difference was observed in the hepatic melanomacrophage density between animals with and without cholelithiasis (Welch two-sample t test, P < 0.001). This study provides an overview on the use of diagnostic imaging modalities to evaluate cholelithiasis and proposes a convenient ultrasound score to standardize monitoring of the condition in L. fallax. It is also the first systematic review of histopathological changes associated with cholelithiasis in this species.
This retrospective study evaluated whether six methods (glutamyltransferase, glutaraldehyde coagulation test, sodium sulfite precipitation test, total serum protein, glucose, and fibrinogen) used to assess passive transfer status in ruminants were predictive of survival of nondomestic Caprinae neonates in a zoological collection. A total of 184 neonates from 10 nondomestic Caprinae species had one or more testing methods performed within 7 d of birth. Results of each test were compared with the clinical condition (alive or dead) at 7, 30, and 90 d of age. Total protein (TP) results were not considered for statistical significance in this study. No statistical correlations between results of the serum gamma glutamyltransferase (GGT), glutaraldehyde coagulation test, or sodium sulfite precipitation test (BOVA-S) and survival at any age were found. A higher glucose level within 7 d of birth was associated with a greater probability of survival. Fibrinogen levels were found to have a strong negative association with survival at 30 and 90 d. Increased glucose concentration was negatively associated with the probability of an infectious cause of mortality and the need for medical intervention. In contrast, increased fibrinogen levels were associated with higher probabilities of infectious death and the need for major medical care. Neonates who were confirmed to have nursed had a lower likelihood of requiring major medical intervention. These findings suggest that glucose and fibrinogen levels are better predictors of neonatal survival in nondomestic Caprinae when compared to the other three tests reviewed in this study. Using survival as an indicator of adequate passive transfer in this group of neonates failed to identify a gold standard of diagnosis of failure of passive transfer, so more than one diagnostic test should be utilized.
Valentina Caliendo, Marian Mensink, Lineke Begeman, Carmen Embregts, Monique de Vrijer, André De Baerdemaeker, Rachel Scheuer, Oanh Vuong, Ron A.M. Fouchier, Thijs Kuiken
Since the emergence of the Goose/Guangdong H5 lineage in 1996 and spillover of highly pathogenic avian influenza (HPAI) from poultry to wild birds, outbreaks have become increasingly frequent in wild birds. The latest outbreak in the Netherlands occurred in the fall-winter of 2020–2021 and was linked to incursions of HPAI H5N8 virus. During the larger national outbreak, wild birds in rehabilitation center “Vogelklas Karel Schot (VKS)” in Rotterdam presented with clinical signs compatible with HPAI, including head shaking, torticollis, and abnormal gait. During an epidemiologic investigation at VKS, water samples from the pools in the enclosures and oropharyngeal and cloacal swabs from 128 birds of different species were analyzed for the presence of H5N8 virus. Forty-five birds and the pool water tested positive for the virus. The outbreak at VKS was likely introduced by one or more infected geese (Anser anser, Anser anser domesticus, Branta leucopsis), after which the virus spread via pool water and with the relocation of infected birds within the center. In principle, such outbreaks are preventable. Recent updates about HPAI to provide guidance to help avoid future incursions of HPAI into wildlife rescue centers are reported.
Protozoal and bacterial vector-borne infections are frequently diagnosed in domestic felids. However, with the exception of Mycoplasma haemofelis and Cytauxzoon felis, their occurrence in managed nondomestic felids housed in the United States is largely unknown. Following a case in February 2020 of fulminant cytauxzoonosis in an African lion (Panthera leo), EDTA–whole blood samples were collected opportunistically from February 2020 through June 2020 from 34 adult tigers (Panthera tigris) and eight adult African lions from the same sanctuary in eastern Tennessee as well as 14 adult tigers from a zoo in southern Oklahoma. Samples were analyzed for Cytauxzoon felis, Bartonella spp., hemotropic Mycoplasma, Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp., and Hepatozoon spp. DNA by PCR amplification. All animals were asymptomatic at the time of collection. None of the Oklahoma animals were positive for vector-borne organisms, but these pathogens were detected in tigers at the Tennessee facility, including Cytauxzoon felis (11.8%), “Candidatus Mycoplasma haemominutum” (5.9%), and Ehrlichia ewingii (2.9%). During the study period, two animals developed clinical signs of cytauxzoonosis and were assessed for vector-borne infections as part of their diagnostic evaluation. This study documents the presence of tick-borne diseases in managed nondomestic felids in the southeastern United States and underscores that ectoparasite control measures should be practiced to minimize exposure of carnivores in managed care.
Aspergillosis remains a difficult disease to diagnose antemortem in many species, especially avian species. In the present study, banked plasma samples from various avian species were examined for gliotoxin (GT), which is a recognized key virulence factor produced during the replication of Aspergillus species hyphae and a secondary metabolite bis(methyl)gliotoxin (bmGT). Initially, liquid chromatography–tandem mass spectrometry methods for detecting GT and bmGT were validated in a controlled model using sera obtained from rats experimentally infected with Aspergillus fumigatus. The minimum detection level for both measurements was determined to be 3 ng/ml, and the assay was found to be accurate and reliable. As proof of concept, GT was detected in 85.7% (30/35) of the samples obtained from birds with confirmed aspergillosis and in 60.7% (17/28) of samples from birds with probable infection but only in one of those from clinically normal birds (1/119). None of the birds were positive for bmGT. Repeated measures from birds under treatment suggests results may have prognostic value. Further studies are needed to implement quantitative methods and to determine the utility of this test in surveillance screening in addition to its use as a diagnostic test in birds with suspected aspergillosis.
Infection with systemic Isospora species (systemic isosporiasis [SI]) is common in passerine birds and may cause substantial mortality in zoological collections. Ten years of postmortem records of 26 species of captive, nonnative passerine birds maintained at the Zoological Society of London, London Zoo, plus seven free-ranging species found dead within the zoo, were reviewed to assess cause of death and occurrence of SI (presence of merozoites in tissue impression smears and/or polymerase chain reaction [PCR] testing for Isospora DNA). The records of 287 juveniles and adults were reviewed, of which 161 had SI test results. The most common cause of death was physical (trauma, predation, drowning, and hypothermia), diagnosed in 39.0% of cases. Virulent SI was considered the cause of death in only nine individuals from five species (3.1% of all cases, 5.6% of tested birds). However, merozoites were recorded in 36.0% of the 150 individuals examined cytologically (representing 18 of the 33 species), while 45.3% of 53 spleen samples (14 species) were positive for Isospora DNA. Test agreement for the 42 birds tested by both methods was 69.0%. Assuming that the PCR result was correct in these, 37.9% of the 161 birds (21 species) were positive for SI at the time of death. These figures might underestimate prevalence because of poor DNA preservation and low numbers of individuals of some species tested. Eight new 28S rDNA sequences and 12 new internal transcriber spacer 1/2 sequences were amplified. Sequences from individuals of the same host species clustered together, suggesting a single Isospora species, and there was no evidence of overlap among hosts. These results confirm that systemic infection with Isospora species in zoo passerines is generally of low pathogenicity and most likely coevolved with their hosts. Severe disease may occur, however, with overwhelming exposure, secondary to immunosuppression, or following coinfection with another pathogen.
Red pandas (Ailurus fulgens) are a globally endangered small carnivoran species and subjects of a robust ex situ conservation effort that includes animals housed in zoos. In 2018, red panda amdoparvovirus (RPAV) was discovered by metagenomics analyses of tissues from two geriatric red pandas, and in one case it was associated with significant lesions. Because RPAV was discovered in a single zoo cohort, it was unclear whether these infections represented a widely distributed, enzootic virus of red pandas or a localized ‘spillover’ from a different host species into this collection. The first goal of this study was to estimate the prevalence of RPAV in US zoos. The authors amplified RPAV from feces of 104 individual red pandas from 37 US zoos, and the virus was detected in 52/104 samples (50.0%). Next, to establish persistence of infection in individual animals, the authors tested serial samples in a single cohort over a 4.5-yr period, and virus was consistently shed by infected animals throughout the sampling period. Finally, full viral coding sequences were amplified and sequenced from three cases, and partial sequences of both the nonstructural and capsid genes were obtained for an additional 19 cases. RPAV is a genetically diverse but monophyletic viral species, and multiple viral lineages are present in US zoo–housed red pandas. The authors do not know how red pandas were originally infected, but RPAV is very common in red pandas in the United States, and infections are persistent—presumably for the lifetime of the animal.
Zahidah I. Zeid, Tom Hughes, Mei-Ho Lee, Jimmy Lee, Reza Tarmizi, Fernandes Opook, Alexter Japrin, Mohamed S.M. Azian, Muhammad Y. Wazlan, Nor A. Aziz, Ronald H.M. Tinggu, Emilly Sion, Siew T. Wong, Jum R.A. Sukor, Jonathan H. Epstein, Peter Daszak
The Bornean sun bear (Helarctos malayanus euryspilus) is the smallest subspecies of sun bear. Their numbers are declining, and more research is needed to better understand their health and biology. Forty-four bears housed at the Bornean Sun Bear Conservation Centre (BSBCC) in Sabah, Malaysia, were screened for known and novel viruses in November 2018. Ursid γ-herpesvirus type 1 (UrHV-1) is a herpesvirus that has been detected from swab samples of clinically healthy sun bears and biopsy samples of oral squamous cell carcinoma in sun bears. We detected an UrHV-1–related virus from throat and rectal swabs by molecular viral screening in samples from 15.9% of the sun bears at BSBCC. None of the bears with the UrHV-1–related virus in this study had oral lesions. There is no known report of UrHV-1 detection in the wild sun bear population, and its association with oral squamous cell carcinoma is not fully understood. Finding an UrHV-1–related virus in a rehabilitation center is a concern because conditions in captivity may contribute to spreading this virus, and there is the potential of introducing it into wild populations when a bear is released. This study demonstrates an urgent need to carry out similar surveillance for sun bears in captivity as well as those in the wild, to better understand the impact of captivity on the prevalence and spread of UrHV-1–related viruses. Positive bears also should be monitored for oral lesions to better understand whether there is a causal relationship.
Melinda A. Gorges, Kassandra M. Martinez, Nicholas F. Labriola, Brianne E. Phillips, Larry Shane Christian, Emile P. Chen, Lysa P. Posner, Gregory A. Lewbart, Daniel S. Dombrowski
The moon jellyfish (Aurelia aurita) is a scyphozoan frequently maintained in public and private aquaria. Little research has been conducted to investigate the effects of various drugs, such as anesthetics, in this species. Tricaine methanesulfonate (MS-222), a common immersion anesthetic for fish and amphibians, was evaluated in a managed population of moon jellyfish. Twenty-four clinically healthy jellyfish were assigned into three groups of eight for trials of 0.3 g/L MS-222 (low concentration [LC]), 0.6 g/L MS-222 (high concentration [HC]), and a saltwater control. The goal was to evaluate the effects of MS-222 administration on moon jellyfish movement and response to stimuli. Movement and response to stimuli were measured via rocking and probe stimulus tests and observations of bell contraction quality and body tone. These tests were performed at baseline and throughout both drug exposure and recovery periods. A threshold drug effect was defined based on systematic scoring criteria. Additionally, elastomer tags were administered to four of eight animals in each MS-222 group to evaluate response to tag placement after drug exposure. Threshold drug effect was achieved in six of eight individuals in the LC group and eight of eight individuals in the HC group. The LC group had median threshold and recovery times of 12.2 and 10.1 min, respectively, while the HC group had median threshold and recovery times of 4.0 and 19.9 min, respectively. The HC group had significantly faster time to threshold drug effect (P < 0.001) and longer recovery times (P= 0.005) than the LC group. In both the LC and HC tagged group, three of four jellyfish had no reaction to tag placement. All animals recovered uneventfully, and there were no mortalities. MS-222 at 0.3 and 0.6 g/L decreased movement and response to stimuli in moon jellyfish.
Alfaxalone, a neuroactive steroid with anesthetic properties, is considered safe when used alone or in combination with other drugs for anesthesia at recommended species doses, and its use has been studied in numerous species. The objective of this study was to assess the pharmacokinetics and pharmacodynamics of IM alfaxalone in Indian peafowl (Pavo cristatus; hereafter peafowl). Eight female peafowl from one zoological institution were used. A control blood sample was obtained before administration of either 10 mg/kg (n = 4) or 20 mg/kg (n = 4) alfaxalone. Blood was collected at 5, 10, 15, 30, and 60 min after alfaxalone injection, with monitoring of sedation score, heart rate, and respiratory rate at each time point. Four peahens receiving a 10 mg/ kg dose had subjectively smoother inductions and recoveries, although sedation level was generally scored as low, with no adverse reactions noted. They were considered fully recovered by the 60-min postinjection time point, although measurable alfaxalone plasma concentrations remained present. Four peahens receiving 20 mg/kg IM experienced adverse effects including seizure-like episodes and hypersensitivity to stimuli throughout the study. This dosing group experienced prolonged recoveries consistent with high plasma concentrations (>3,000 ng/ml). Based on these results, use of 20 mg/kg IM alfaxalone as the sole anesthetic agent is not recommended in this species. Further studies should determine whether alfaxalone in conjunction with other anesthetic or analgesic agents could provide better sedation and smoother induction and recovery for peafowl and to assess the pharmacokinetics and pharmacodynamics of alfaxalone in other avian species.
Chemical sedation of fish is often used during scientific research to reduce stress and risk of injury. Electric fish handling gloves (EFHG) have been proposed as a no-residue alternative to chemical immobilization. However, the impact of handling fish with EFHG on their physiology remains relatively poorly studied. Stress markers (plasma cortisol and lactate) and the impact on muscular integrity (plasma creatine kinase and histopathology of skeletal muscles) were assessed in brook trout (Salvelinus fontinalis) subjected to one of three immobilization methods (manual restraint, EFHG [10–16 mV], and buffered 60 mg/L tricaine methanesulfonate [MS-222] immersion anesthesia) for 1- and 5-min restraints. Plasma lactate levels were significantly higher 2 h postrestraint for all experimental groups (+242%; P < 0.001), but the magnitude of these increases was significantly lower in the MS-222 groups (P ≤ 0.0002). Plasma creatine kinase concentrations significantly increased 2 h postrestraint for the EFHG groups (+92%, P ≤ 0.0061), and this increase was significantly higher than in the MS-222 group for the 1-min restraint (P = 0.0031). Although there was no significant difference between the restraining methods tested at all time points for plasma cortisol, the EFHG and manual restraint groups showed a linear and statistically significant increase after the initial 5-min restraint, whereas all other groups presented an expected bell-shaped profile with a plasma peak 1 h after the initial restraint. One fish in the manual restraint group died during the 5-min restraint, and two fish from the EFHG group were euthanized at the end of this protocol because of buoyancy anomalies. All other fish from the 5-min restraint group developed cutaneous saprolegniasis 3 wk after the experiments. The evaluation of the safety of EFHG in brook trout showed similar impacts on the variables tested compared with manual restraint for 1- and 5-min immobilizations.
Effective management of articular injuries in avian species is a known and frequent challenge. Potential treatments include many domestic animal therapeutics, such as Adequan®, which is used widely in dogs and horses. However, clinical reports have described hemorrhagic diatheses in a variety of avian species treated with varying doses and administration frequency of Adequan. This study investigated the hypocoagulability associated with parenteral administration of Adequan in avian species. Following a pilot dosing study in domestic chickens (Gallus gallus), citrated plasma from Chilean flamingos (Phoenicopterus chilensis) (n = 42) was spiked with Adequan to represent three dosing regimens (1 mg/kg, 5 mg/kg, 10 mg/kg). The fibrinogen content of plasma samples was determined and thrombin-clotting times (TCTs) were compared for the untreated (control) and spiked flamingo samples. The TCT for control and 1-mg/kg spiked plasma were not significantly different; however, both 5 mg/kg and 10 mg/kg spiked samples demonstrated significantly prolonged TCT (P-value < 0.0001) indicating hypocoagulability. These results support that Adequan given parenterally at 1 mg/kg can be utilized safely in clinical case management as an adjunctive treatment for osteoarthritis in flamingos and potentially other avian species.
Thromboelastography (TEG) provides a global assessment of hemostasis and fibrinolysis and has broad applications to identify and monitor coagulation dysfunction in veterinary patients. Although alpacas are susceptible to a wide variety of coagulopathies, the assessment of TEG has not been reported in clinically healthy alpacas to date. The purpose of this study was to evaluate the analytical performance of recombinant human tissue factor (rhTF)– and kaolin-activated TEG and to establish reference intervals for TEG parameters (reaction [R] and clotting [K] times, angle [α], maximum amplitude [MA], and shear elastic modulus [G]) in healthy, adult alpacas. Kaolin and rhTF-activated TEG were performed using citrated whole blood samples from 20 clinically healthy, nonpregnant, adult Huacaya alpacas each after 30 min of sample storage at room temperature. Six individuals of a related species, dromedary camels, were also sampled for comparative purposes. All data were presented descriptively, assessed for normality, and compared using either independent-sample t tests or Mann-Whitney U tests, with P ≤ 0.05 considered significant. Reference intervals were calculated using a robust method and Box-Cox–transformed data. Mean TEG values (reference intervals) were determined for rhTF-activated TEG as follows: R 6.99 min (3.41–12.71), K 3.43 min (1.61–6.42), α 48.51° (27.21–67.38), MA 52.05 mm (21.53–65.92), and G 5.71 kdyn/cm2 (1.87–9.60), while mean values (reference intervals) for kaolin-activated TEG included R 7.72 min (4.48–11.43), K 4.24 min (2.03–9.20), α 45.06° (23.66–64.20), MA 52.18 mm (33.49–66.63), and G 5.78 kdyn/cm2 (NR–9.66). None of the measured TEG values differed significantly between activators, suggesting that activator choice may have a limited effect on TEG parameters in healthy alpacas. TEG results in alpacas were comparable to those of dromedary camels. These results will thus provide a useful starting point in the evaluation of hemostasis in adult camelids.
Objective disease markers in the southern white rhinoceros (Ceratotherium simum simum) are in high demand. In the field, such markers are typically needed to decide whether a captured white rhinoceros is fit to cope with quarantine, transport, or both. Captive white rhinoceros have a need for unbiased biomarkers for early detection of disease. Acute phase proteins, including haptoglobin, are proteins that significantly change their plasma concentration in response to tissue perturbation or inflammation, such as that occurring during infection or neoplastic disease. Acute phase proteins are well known diagnostic tools in both human and veterinary medicine. In this study, an ELISA with commercially available anti-human haptoglobin antibodies for quantification of haptoglobin in white rhinoceros serum was developed. The validity of the haptoglobin assay and haptoglobin as a biomarker of disease was investigated with the use of serum samples from both captive and free-ranging animals with a well-described health status. The assay was precise (intra-assay and interassay reproducibility were 5.0% and 13.1%, respectively) and reliably quantified white rhinoceros haptoglobin serum concentrations consuming low volumes of sample. The assay was sensitive to the presence of free hemoglobin in the sample at levels corresponding to a visibly hemolyzed sample. Haptoglobin was readily measurable, baseline levels (in white rhinoceros with no clinical signs of disease) did not differ between genders, and a significant increase was seen in captive as well as in free-ranging white rhinoceros with inflammatory disease. Thus, haptoglobin is a positive acute phase protein in southern white rhinoceros with potential for use as an objective marker of disease.
Elasmobranchs are popular display animals in public aquaria and zoos, but medical management gaps remain in the understanding of the pharmacokinetics of analgesics and pain management in these species. Meloxicam is a nonsteroidal anti-inflammatory drug that has been evaluated intravenously and intramuscularly in teleosts, but has yet to be studied in any elasmobranch species. The pharmacokinetics of meloxicam were determined in 17 yellow stingrays (Urobatis jamaicensis). All stingrays were determined to be healthy from complete physical examinations and baseline bloodwork performed prior to study inclusion. A single dose of 1 mg/kg meloxicam intramuscularly was administered to all rays, followed by a 2 mg/kg oral dose after an 8 wk washout period. Blood samples were collected from the mesopterygial vein at baseline and nine time points up to 96 h after administration of meloxicam. Plasma concentrations were determined using reversed-phase high-performance liquid chromatography. Pharmacokinetic analysis was performed using a noncompartmental technique. The mean peak plasma concentrations for intramuscular and oral meloxicam were 1.29 and 0.42 µg/ml, respectively. The mean terminal half-lives of meloxicam after intramuscular and oral administration were 5.75 and 15.46 h, respectively. Based on these findings, the recommended meloxicam dosage and frequency for yellow stingrays is 2 mg/kg orally once daily. Due to rapid elimination with the intramuscular administration, maintaining clinically relevant plasma concentrations may be difficult using this route. Further studies are needed to determine multidose pharmacokinetics of meloxicam in yellow stingrays, as well as single-dose and multidose pharmacokinetics in other elasmobranch species.
Katja Schönbächler, Helmut Segner, Bénédicte Amphimaque, Brian Friker, Andreas Hofer, Barbara Lange, Martina Stirn, Nikola Pantchev, Francesco C. Origgi, Stefan Hoby
The highly endangered European pond turtle (Emys orbicularis) was reintroduced in Switzerland in 2010. Up until 2019, no routine medical examinations have been carried out prior to its release or during recapture events. The aim of this study was to assess the health status of captive and free-living Emys orbicularis populations in Switzerland, taking into account the most important and frequently occurring health threats to freshwater turtles. A total of 141 European pond turtles, including captive (n = 89) and free-living (n = 52) individuals, underwent clinical examination (n = 136), choanal and cloacal swab collection for microbiology investigation (n = 140), blood sampling (n = 121), fecal examination for parasitology (n = 92), radiography (n = 84), and ultrasound (n = 46). Microbiology investigation included conventional PCR for herpesvirus, ranavirus, and Mycoplasma spp. Blood was used for the establishment of reference values for hematocrit, leukocyte count, and differential blood count as well as for biochemistry parameters tested with the VetScan VS2. An emydid Mycoplasma was detected in 40% (n = 56/140; 95%CI: 31.82–48.61%) of the turtles, including one individual with upper respiratory signs. Four animals positive for Mycoplasma arrived dead or were euthanized during the study period. Their necropsies revealed no evidence of respiratory disease. No ranavirus or herpesvirus was detected in any of the tested turtles. Two presumptively fatal infections with spirorchiid trematodes were reported during the study period. Endoparasites were detected in only 7.94% of the samples examined. This study provides comprehensive data on the current health status of the largest sample size of captive and free-living populations of Emys orbicularis ever assessed to date and serves as a baseline for future research investigations and management recommendations in this species.
Although the evaluation of hematologic and biochemical parameters is a well-established diagnostic tool in vertebrate medicine, comprehensive understanding of these parameters in invertebrate species is lacking. This study provides baseline hemocyte concentrations and biochemistry values for a population of managed Japanese spider crabs (JSC; Macrocheira kaempferi) housed at six different public aquariums. The methodology for obtaining diagnostic hemolymph samples is described. Distinct hemocyte types were identified, including hyaline cells, semigranulocytes, and granulocytes, with hyaline cells as the predominant type. Correlates to exam findings and environmental parameters were evaluated and included higher absolute semigranulocyte counts (r = 0.65, P = 0.020) and triglyceride levels (r = 0.44, P = 0.014) in JSC with exoskeletal lesions; higher total protein (mean = 5.93 g/dl, P = 0.028), cholesterol (median = 18.5 mg/dl, P = 0.018), triglyceride (median = 15.5 mg/dl, P = 0.002), and amylase (median = 243 U/L, P = 0.013) in nonmolting JSC compared with JSC that have previously molted since acquisition (total protein mean = 4.83 g/dl, cholesterol median = 14 mg/dl, triglyceride median = 6.4 mg/dl, and amylase median = 131 U/L); and lower relative and absolute granulocyte counts (mean = 8.83% P = 0.030, median = 1,162 cells/µl P = 0.006, respectively) and higher albumin (median = 1.35 g/dl, P = 0.031) in JSC housed at facilities that used ozone sterilization. The data presented serve as a foundation for understanding basic clinical parameters in JSC hemolymph, as well as the potential influence of environmental stressors on those parameters.
Reference intervals for clinical chemistry analytes are important for the interpretation of blood results, but reference intervals available in journal articles for specific flamingo collections are often based on a small number of individuals or are only available for a limited number of analytes. Differences in husbandry and nutritional conditions may impact reference intervals. The goal of this study was to establish reference intervals from a mixed population of greater (Phoenicopterus roseus) and American (Phoenicopterus ruber) flamingos when husbandry parameters are equal and to compare the results between the different species, sexes, and age groups in order to evaluate differences between these factors. Lithium-heparinized plasma samples from 93 animals were analyzed for the following: amylase, cholinesterase (CHE), bile acids (BA), total protein (TP), glutamate dehydrogenase (GLDH), aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), uric acid (UA), inorganic phosphorus (P), total calcium (Ca), sodium (Na), and potassium (K). The results show many differences between the species and a few between the different age groups but no differences between males and females of either species. Amylase (P= 0.0010), AST (P= 0.0090), CK (P= 0.0011), and Ca (P= 0.0217) differed significantly between greater and American flamingos. The age group significantly impacted blood levels of CK (P = 0.0299) and LDH (P = 0.0165) in greater flamingos and BA (P = 0.0495), P (P = 0.0422), and Ca (P = 0.0006) in American flamingos. The results show the importance of species-specific reference intervals and the consideration of age-related variations in order to facilitate correct interpretation of blood results.
Black-footed ferrets (Mustela nigripes) are an endangered species in North America that are highly sensitive to canine distemper virus (CDV) infections and any exposure could be devastating to species recovery. The U.S. Fish and Wildlife Service recovery program has safely used a recombinant DNA (rDNA) canarypox-vectored CDV vaccine, Purevax® Ferret Distemper (PFD), to vaccinate black-footed ferrets. Because of a PFD shortage in 2015, an rDNA vaccine labeled for use in dogs, Recombitek® CDV (rCDV), was chosen to vaccinate black-footed ferrets. Our goal was to compare the serum neutralizing (SN) titers after vaccination of 17 captive and 18 wild black-footed ferrets with rCDV or PFD, respectively, considering ≥1:128 as a protective titer. Both vaccines produced comparable 1 yr postvaccination protective titers in captive and wild black-footed ferrets. In wild black-footed ferrets, one PFD vaccination produced SN titers similar to two PFD vaccinations at 1 yr postvaccination. One year after vaccination with rCDV, SN titers in captive black-footed ferrets were higher than in wild ferrets. These results indicate rCDV may be an effective alternative CDV vaccine in captive black-footed ferrets and PFD should be prioritized for wild ferrets because one dose was effective for animals that can be difficult to recapture.
Renal disease is well documented in nondomestic felids and is monitored and diagnosed by serum concentration of blood urea nitrogen, creatinine, and phosphorous. Symmetric dimethylarginine (SDMA) has proven to be an earlier and more sensitive biomarker for the assessment of glomerular filtration rate. Although SDMA is commonly measured in nondomestic felids, information concerning the validity of the assay is lacking. The purpose of the study was to perform a method comparison between high-throughput immunoassay and the reference method, liquid chromatography–tandem mass spectrometry (LC-MS/MS), to quantify SDMA concentrations in tiger blood samples. Concentrations of SDMA were measured for 81 individual tiger samples. The SDMA immunoassay demonstrated excellent correlation to the LC-MS/MS reference method. A Passing and Bablok linear regression analysis had a slope of 1.03 (95% CI, 0.99–1.11), an intercept of 1.64 (95% CI, 0.46–2.34), and a Pearson R= 0.99. The mean bias was 1.53 µg/dl (95% CI, 0.63–2.42 µg/dl), and the limit of agreement was ±7.96 µg/dl. The degree of bias is within established acceptance criteria of 1–3 µg/dl for the immunoassay. Although this study provides good evidence of the utility of the immunoassay to measure SDMA in tiger serum and plasma, further assay validation is recommended.
Published research on the health status of wild bald eagles (Haliaeetus leucocephalus) in Eastern Canada is limited. This study examined the health status of 53 nestling bald eagles on Prince Edward Island in the summers of 2012 and 2013, including biochemistry reference intervals, a summary of hematological parameters, and background lead accumulation. Baseline lead levels of greater than 0.02 ppm were noted in nine nestlings, and the odds of lead accumulation decreased with increased weight and body condition score. Biochemical reference intervals were developed using a method outlined by the American Society of Veterinary Clinical Pathology. These intervals can vary between populations based on age and geographic area. These findings will therefore be useful in further research and clinical cases within this population, as well as further research into subclinical lead accumulation in growing raptors.
The Turtle Rescue Team (TRT) at North Carolina State University College of Veterinary Medicine currently sees over 500 chelonian cases a year. Each turtle receives a thorough physical examination as the main evaluation of its health because further diagnostics are cost prohibitive in a donation-based facility. Thus, there is a need to further understand how a few economical diagnostics can evaluate the prognosis of chelonians. The objectives were to evaluate lactate, PCV, total solids, glucose, temperature, and HR as prognostic indicators in multiple chelonian species and to determine how the change in lactate concentrations over a 24 h period affects prognosis. Eastern box turtles (Terrapene carolina carolina), yellow-bellied sliders (Trachemys scripta scripta), river cooters (Pseudemys concinna), painted turtles (Chrysemys picta), and common snapping turtles (Chelydra serpentina) were evaluated because of prevalence of presentation to the TRT, differences in habitat, and differences in physiology. Hyperlactatemia, an increased lactate over 24 h when the initial lactate concentration was greater than 5 mM/L, and anemia were determined to be significant prognostic indicators of mortality for all turtles, and hypoglycemia was a significant prognostic indicator of mortality for male turtles (P < 0.05).
María de Jesús Rovirosa-Hernández, Juan Francisco Rodríguez-Landa, Francisco García-Orduña, Eduardo A. Gazol-Patiño, Omar Lagunes-Merino, Paola Heman-Bozadas, Abril de los Ángeles Aguilar-Tirado
The determination of health status in spider monkeys (Ateles geoffroyi) that are held in managed care requires periodic evaluation. The present study obtained baseline data on hematologic values, body weight, and length in A. geoffroyi. A total of 118 individuals from three housing centers were included in the study and grouped into two categories by age (92 adults, 26 juveniles) and sex (46 males, 72 females). Body weight, red blood cell (RBC) counts, and hemoglobin counts were significantly higher in adult males than in adult females. No differences in length were found between sex and age groups. The present findings indicate that hematologic values by age and sex in A. geoffroyi are similar to those reported in other spider monkey populations in both managed care and the wild. These results will be useful as a reference for young individuals and adults in future studies of the health of this species.
André Luiz Mota da Costa, Emanuel Vitor Pereira Appolonio, Luiz Henrique Lima de Mattos, Gustavo dos Santos Rosa, Leandro Silva Reis, Beatriz Maccari Silva, Matheus Vasconcellos, Rodrigo Hidalgo Friciello Teixeira, Ana Liz Garcia Alves
Tapirs are endangered terrestrial mammals that inhabit several continents. They have anatomical similarities to horses, sharing a common ancestral lineage. This article reports the case of a 14-yr-old female lowland tapir (Tapirus terrestris) presented for intermittent lameness due to upward fixation of the patella causing extension of the limb in the caudal phase of the stride. Medial patellar desmotomy was performed under general anesthesia, correcting the problem. To date there are no reports of this condition or treatment recommendations in tapirs. An anatomical study including stifle dissection, advanced MRI, and CT was performed in a separate lowland tapir. According to the clinical case and the anatomical findings in the other lowland tapir, upward fixation of the patella may occur in the tapir, although the anatomy varies slightly from that of the horse. Because the lowland tapir does not have parapatellar cartilage or as large of a medial patellar ligament or medial trochlea of the distal femur compared to the horse, more severe disease secondary to complete or persistent upward fixation of the patella may not occur in tapirs. Rather, mild forms of the disease associated with intermittent upward fixation of the patella or delayed patellar release appear more likely in the tapir.
In 2019, two wild-caught adult female eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) received a preshipment examination and were individually swabbed for chytrid testing via quantitative polymerase chain reaction (qPCR). Physical examination was unremarkable. Both females tested positive for Batrachochytrium dendrobatidis (Bd) and negative for B. salamandrivorans (Bsal). A course of terbinafine hydrochloride 1% in alcohol was administered in a 0.005% treatment bath for 5 min once daily for 5 d. Both animals were individually retested 1, 3, and 4 wk after treatment using qPCR. All post-treatment samples were negative for Bd and Bsal. This report represents the first successful treatment with terbinafine hydrochloride 1% in alcohol to eliminate subclinical Bd infection in eastern hellbenders and underlines the importance of preshipment testing for chytrid in all amphibians being transferred to new facilities or released into the wild as a means to minimize risk of disease introduction via subclinically infected individuals.
Maya L. Iyer, Christine M. Molter, Joseph P. Flanagan, Kendra L. Bauer, Rob Bernardy, Daryl Hoffman, Lily Parkinson, Benjamin M. Brainard, Tierra Smiley Evans, Taylor Pursell, Paul D. Ling
Novel diagnostic and therapeutic methods were utilized in the successful management of severe elephant endotheliotropic herpesvirus hemorrhagic disease (EEHV-HD) in a 1.9-yr-old captive Asian elephant (Elephas maximus). High levels of EEHV1A viremia were detected for 12 d. In addition to established EEHV treatments, therapies included famciclovir-fortified elephant whole blood and plasma, mesenchymal stem cells harvested from elephant umbilical tissue, and aminocaproic acid. Testing conducted to examine the effects of EEHV infection on hemostasis suggested marked intravascular coagulation with decreased plasminogen activity and increased D-dimer concentrations. Thromboelastography was used to assess the efficacy of aminocaproic acid and demonstrated hypofibrinolysis on samples taken after drug administration, as compared with samples from healthy adult Asian elephants. A serological assay for a novel EEHV1A-specific antibody marker (E52) was developed due to lack of seroconversion to a previously established EEHV1A-specific antibody marker (ORFQ) and showed a sustained increase after EEHV-HD illness.
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