CLINICAL SIGNS, ANTEMORTEM DIAGNOSTICS, AND PATHOLOGICAL FINDINGS ASSOCIATED WITH MYCOBACTERIUM AVIUM SUBSPECIES PARATUBERCULOSIS INFECTION IN MISHMI TAKIN (BUDORCAS TAXICOLOR TAXICOLOR) Adam D. Naylor, Douglas Richardson, Morag Sellar, Jane Harley, Adrian W. Philbey, Simon J. Girling 49(2), 412-419, (1 June 2018) https://doi.org/10.1638/2017-0215.1

Mycobacterium avium subspecies paratuberculosis (MAP) is a cause of contagious and typically fatal enteric disease, primarily affecting ruminant and pseudoruminant species. During a MAP outbreak in a captive collection, six of nine adult Mishmi takin (Budorcas taxicolor taxicolor) showed marked weight loss over 1-3 mo, followed by an acute deterioration. Fecal culture and microscopy failed to identify MAP shedding. Necropsy findings included grossly normal intestines and marked enlargement of mesenteric lymph nodes. Histological findings included multibacillary granulomatous enteritis, mesenteric lymphadenitis, and periportal hepatitis. MAP was confirmed by culture of intestinal and lymph node tissues from the index case. Results of antemortem serological testing using an indirect ELISA (ID SCREEN® Paratuberculosis Indirect) were corroborated by findings at necropsy or survival of the outbreak. Mishmi takin appear to show high MAP susceptibility and a rapid disease course compared with domestic ruminant species.

Case Series Mycobacterium avium ssp. Paratuberculosis (MAP) is etiologic agent of Johnne’s disease in

domestic ruminants. Cases here are first report of MAP in Mishmi takin 6/9 Mishmi takin developed disease consistent with MAP and were euthanized during disease

outbreak at UK zoo Weight loss seen in all affected takin Behavioral changes seen in all takin 1-3 wk prior to euthanasia- self isolation, hyporexia,

malaise, tensing abdomen Intermittent diarrhea seen in 2/6 cases

Diagnostics - No acid-fast bacteria seen on pooled fecal samples and all MAP cultures negative 5/9 of the herd were + on ELISA serologic testing

Necropsy of euthanized animals - poor BCS, enalrged pale tan mesenteric LN with yellow-white foci, intestines were grossly unremarkable

Histo- mesenteric LN had granulomatous inflammation, lymphocytic-plasmacytic inflammation in intestines and periportal hepatitis, with acid-fast bacilli in mesenteric LN

Positive cultures achieved off ileum and mesenteric LN Outbreak included MAP-like lesions in yaks, markhor, European bison, reindeer, red deer, takin

MAP infections confirmed with tissue culture or PCR Remaining 3 takin vaccinated with heat-inactivated MAP vaccine

1 died of other cause since vaccination, no histo evidence of MAPTakeaways

Case series reporting MAP in Mishmi takin. 6/6 affected takin showed weight loss. Diarrhea was present intermittently and late in disease process. No antemortem cultures or acid-fast fecal stains were +, but cultures and histo post-mortem confirmed MAP

6/6 affected animals showed serologic response antemortem, with clinical animals being more strongly positive

MAP should be considered for takin with weight loss, behavioral changes, diarrhea

INTERDIGITAL NECROBACILLOSIS ASSOCIATED WITH TRUEPERELLA PYOGENES IN GOITERED GAZELLES (GAZELLA SUBGUTTUROSA) Göksen Çeçen, Esra K. Buyukcangaz, Ülke G. Çalışkan, Taci I. Cangul, Ezgi Akdesir 49(2), 429-434, (1 June 2018) https://doi.org/10.1638/2017-0050.1

This study presents the clinical findings, treatment results, and gross pathology of Trueperella pyogenes–associated interdigital necrobacillosis in captive goitered gazelles (Gazella subgutturosa). Four male and two female gazelles presented with weight loss, front limb swelling, and persistent lameness despite antibiotic treatment. The animals were reluctant to bear weight on the affected limbs, and the diagnosis of interdigital necrobacillosis was made based on physical exam, bacteriologic evaluation, and radiographic imaging. In all cases, the interdigital skin and subcutaneous tissues were affected in both forelimbs. Exungulation (loss of the hoof) occurred in one female gazelle. Despite aggressive topical and systemic treatment, the two female gazelles died. Gross pathology detected abscesses in the heart, lung, and liver. T. pyogenes was isolated as the major pathogen. This is the first report of interdigital necrobacillosis caused by T. pyogenes in Gazella subgutturosa.Introduction:

Interdigital necrobacillosis (foot rot): contagious bacterial hoof disease that occurs in domestic and nondomestic ungulates

caused by complex of bacteria: Fusobacterium necrophorum, Dichelobacter nodosus, Trueperella pyogenes, Porphyromonas levii, Prevotella intermedia, and others

T. pyogenes: gram-positive, nonmotile, nonspore- forming, short, rod-shaped bacterium GOAL: present clinical findings, tx results, and gross pathology of foot rot in captive goitered

gazelles All 6 gazelle presented over 2 month period with weight loss, forelimb swelling and lameness One female had putrid necrotic tissue from coronary band to carpus; some respiratory signs DX: blood work normal, no bony changes on rads; culture: isolated Trueperella pyogenes Tx: debrided wounds PRN; started with IM lincomycin, but upgraded to IV vancomycin; banamine 2 cases got regional limb perfusion with some kind of crystalline penicillin One female had exungulation (hoof fell off); both females died (pneumonia/septic changes seen) Males recovered without recurrence All animals spent a lot of time in the outside yard during rainy season = predisposing factor

 Take Home: First report identifying T. pyogenes as a cause of interdigital necrobacillosis in goitered gazelles

Grøndahl, C., Andersen-Ranberg, E. U., Mosbacher, J. B., Stelvig, M., Hansen, L. H., & Schmidt, N. M. (2018). Immobilizing muskox (ovibos moschatus) under high arctic conditions. Journal of zoo and wildlife medicine, 49(4), 856-862.

Immobilizing and handling large, free-ranging animals without proper facilities in harsh environmental conditions poses significant challenges. During two field expeditions, a total of 29 female muskoxen (Ovibos moschatus) were immobilized in Northeast Greenland (74°N, 20°E). Fixed doses of immobilizing drugs were used regardless of animal size: 2 mg etorphine, 30 mg xylazine, 0.3 mg medetomidine, and 40 mg ketamine. Physiologic and behavioral monitoring was performed during the second expedition on 15 female muskoxen. The observed heart rates were 35–58 beats/min and respiratory rates were 25–30 breaths/min. Mean arterial pressures measured using oscillometry ranged between 117–142 mmHg. Pulse oximeter readings ranged from 91–98% with oxygen supplementation, nasal end-tidal carbon dioxide values were 24–42 mmHg, and rectal temperature ranged from 38.9–39.6°C. Induction time was

6–8 min, recovery time 2–6 min after reversal, and duration of anesthesia was 50–100 min. This anesthetic regime thus provided reliable immobilization with minimal pathophysiologic alterations.

- Introduction:o Muskoxen studied in NE Greenland, high Arctic, -9 deg C mean annual.

Low temps, high wind speeds, low wind-chill compromises equipment, physical-chemical properties of drugs, and manual dexterity of personnel.

o Free-ranging muskoxen immobilized 2013-2015, goals to develop logistic and drug protocols and evaluate physiologic parameters of animals during immobilization.

- M+M:o Zackenberg Valley, NE Greenland, 2013, 2015.

27 free-ranging female muskoxen. On foot and ski, moved herd towards an elevated area and circled toward

desired location. Darted with DanInject CO2 powered dart gun.

Kept warm by wrapping in bubble wrap with hand warmers.o Fixed doses – 2 mg etorphine, 30 mg xylazine, 0.3 mg medetomidine, 40 mg

ketamine. Approximately 0.01 mg/kg etorphine, 0.15 mg/kg xylazine, 1.5 mcg/kg

medetomidine, 0.2 mg/kg ketamine. Total volume of 1.5 mL in dart, supplemented with sterile water and

ethanol to prevent freezing. Reversed with naltrexone and atimapezole.o Monitored RR, HR, O2 sat, ETCO2, noninvasive BP, rectal temp. Monitors

wrapped in bubble wrap and hand warmers or warmed fluid bags to maximize normal function.

o Venipuncture – jugular vein, preheparinized syringe, EDTA and serum tubes. Arterial measurement not attempted due to need to remove hair and risk of

frostbite. Serum cortisol measured.o Parameters evaluated by calculating mean across muskoxen for each 10-min

interval post-darting, compared to previously reported ref ranges.- Results:

o All immobilizations were successful, no complications. Induction – uncoordination, dog sitting, relaxing into sternal recumbency. ~1/3 animals supplemented with drugs via pole syringe. 19/29 were partially reversed with atipamezole and doxapram, too deep. Full reversals were unremarkable, walking away 2-6 min after.

o Mean HR 35-58 BPM, RR 25-30 BPM, BP 117-142 mmHg, pulse ox 91-98%, mean and end-tidal CO2 24-42 mmHg, T 102-103.3 deg F.

o Mean cortisol 76nmol/L in 2013 and 108 nmol/L in 2015.- Discussion:

o This drug combo provides high likelihood of success with minimal risk of complications.

Induction times comparable to potent opioid anesthesia regimes.

Immobilization up to 100 minutes without supplementation. One bull was accidentally darted and was also adequately immobilized. Ability to deepen by supplementation or lighten with partial reversal

allowed for excellent manipulation of depth.o Physiologic parameters.

Most fell within previously reported reference intervals. HR low normal, likely due to alpha 2 agonist. BP slightly elevated, likely due to combo of alpha 2 agonist and ketamine

and etorphine in ruminants in general. BP slightly elevated vs those reported with butorphanol,

azaperone, medetomidine and opioid medetomidine-immobilized ungulates.

Rectal temps slightly elevated vs reference ranges. Small, increasing trend during anesthesia. Attributed to insulating properties of muskox fur. Avoidance of hyperthermia likely due to careful approach by the

team, animals did not become agitated or run or struggle before darted.

Supplement of oxygen at 1 L/min per 100 kg appeared to yield superior oxygen saturation vs values reported in studies without supplemental O2.

o Serum biochemistry and stress. Cortisol levels markedly lower than those reported for gunshot and

helicopter-immobilized muskoxen. Only moderately elevated. It is recommended to approach muskoxen on foot to reduce level of

physiologic stress.o Using balanced anesthesia technique allows for use of lower doses of potent

opioids and physiologic responses closer to normal range.

Martinez, M. E., Zimmerman, D., Seeley, K. E., Zhang, L., Bapodra, P., & Cianciolo, R. E. (2019). Systemic amyloidosis in a population of pronghorn antelope (Antilocapra americana). Journal of Zoo and Wildlife Medicine, 50(1), 147-158.

Abstract: Thirteen pronghorn antelope (Antilocapra americana) from a single captive herd at the Columbus Zoo and Aquarium underwent complete or partial necropsies between 1997 and 2016. Ten of the 13 animals had histologic evidence of amyloidosis resulting in a 77% prevalence. Histologic and ultrastructural changes were characterized in an attempt to determine the underlying cause of the amyloid. Amyloid detection was performed through histologic examination of hemotoxylin and eosin and Congo red–stained microscopic slides for all 13 animals. Transmission electron microscopy and mass spectrometry was performed on renal tissue from two animals. Pedigree analysis and retrospective investigation into the clinical histories was performed. Histologically, 9/10 animals had amyloid present in the kidneys, 8/10 in the liver, 9/10 in the spleen, 4/10 in the gastrointestinal tract, 3/10 in the adrenal

glands, and 2/10 in the thyroid glands. Transmission electron microscopy demonstrated glomerular deposits consistent with amyloid. Mass spectrometry performed on renal specimens from two animals revealed the presence of serum amyloid A. Eight of the 10 animals diagnosed with systemic amyloidosis had a clinical history of haemonchosis (elevated fecal strongyle count), 5/10 were diagnosed with pneumonia postmortem, and 7/10 had postmortem findings consistent with negative energy balance. Serum amyloid A, and β and γ globulin levels were evaluated in four cases of amyloidosis, and all were within normal ranges for healthy domestic cattle. It was possible that the herd's amyloidosis was associated with a hereditary defect that could be exacerbated by chronic inflammation. However, there was no significant association between the mean degree of relatedness and presence of amyloidosis. In conclusion, systemic amyloidosis in this captive population of pronghorn is common. It is likely reactive and secondary to underlying chronic inflammation caused by haemonchosis and/or pneumonia.

- Introduction:o Pronghorn antelope (Antilocapra americana) – W. North America, only extant member

of Antilocapridae.o Amyloidosis: Disorder of aberrant deposition of misfolded, insoluble fibrils in a beta

pleated sheet configuration. Most common sites of deposition – kidney, liver, spleen.

Adrenals, thyroid glands, nervous system, heart less common. Many precursor proteins can become misfolded and form amyloid deposits –

immunoglobulin fragments, transporters i.e. apolipoproteins, and acute phase proteins i.e. SAA.

Humans – immunoglobulin-derived amyloidosis most common. Domestic animals – reactive systemic amyloidosis from serum amyloid

A. Acute phase protein, produced by hepatocytes in response to proinflammatory cytokines i.e. IL1, IL6, TNFalpha, common sequela to chronic inflammation.

o Also familial amyloidosis i.e. Beagles, Shar Peis, Abyssinian and Siamese cats.

Dx via histology – eosinophilic homogenous proteinaceous material within vessel walls and/or extracellular matrix, demonstrates apple-green birefringence when Congo red-stained slides examined with polarized light.

Typing of the precursor protein can be further investigated. IHC, immune-electron microscopy, and/or mass spectrometry-based

approaches. Amyloidosis has been previously reported in captive wild ungulates – bongo,

gazelle, beira antelope, kudu.- M+M:

o 13 pronghorn antelope, necropsies between 1997-2016, histo slides for retrospective study.

o Sections with suspected amyloid were stained with Congo red. Two cases with renal samples stained with periodic acid-Schiff and Masson’s trichrome, and Jones’s methenamine silver. Evaluated by TEM. Mass spec to ID proteins (SAA).

o Pedigree analysis to determine degree of relatedness between affected animals.

o Haptoglobin, SAA, and immunoglobulin measurements were performed on four cryo-banked sera samples.

- Results/Discussion:o 13 antelopes, 5 mos – 14 yo, 8 M, 5 F. 10 histologic evidence of amyloidosis (77%), range

3-14y, 6M, 4 F. Clinical histories – abnormalities related to systemic amyloidosis included

azotemia (5/10), proteinuria (1/10), hypocalcemia (6/10), hyperphosphatemia (1/10), elevated BUN (1/10). Hypoproteinemia 6/10. Diet – ADF-16 wild herbivore pellets, alfalfa, browse when available.

o Nx findings: Gross – bilateral pale pink renal cortices with some cases with cortical cysts or

multifocal cortical atrophy and fibrosis characterized by 2mm diameter cortical depressions. Two cases, renal cortices pale yellow.

Histo – 9/10 amyloid both kidneys, 8/10 amyloid in liver, 4/10 amyloid in spleen, 3/10 amyloid in GIT, 3/10 in adrenals, 2/10 in thyroid glands.

Dx based on presence of glassy amorphous eosinophilic matrix demonstrating apple-green birefringence when Congo red stain viewed with polarized light.

Special stains consistent with other species – amyloid stained pale pink with PAS, mottled pale blue with Masson’s trichrome, did not take up silver with JMS.

4/9 animals with amyloid in kidneys only had amyloidosis in the medulla, not detectable in glomeruli.

TEM on two cases – marked expansion of the mesangial zones and mild to moderate expansion of the capillary walls by haphazardly arranged nonbranching fibrils.

Liver – amyloid present in space of Disse, most prominent in hepatic cords near portal and/or centrilobular vasculature.

Spleen – amyloid most often around and/or within periphery of periarteriolar lymphoid sheaths extending into the adjacent red pulp.

Adrenal – within zona fasciculate. Thyroid – within follicular basement membrane and interstituim. GI – all 4 cases also had forestomach microabscesses, but two of six with

microabscesses did not have GI amyloidosis.o Amyloidosis typing – SAA and haptoglobin within normal limits.o Investigating cause of pronghorn systemic amyloidosis:

Chronic inflammation – haemonchus with 1,000 EPG or higher 90% of individuals from 1997-2016 (21).

Two animals with amyloidosis without clinical haemonchosis had pneumonia.

5 animals had gross or histo diagnoses of bronchopneumonia, most commonly due to aspiration or bacteremia.

70% of pronghorn with amyloidosis had gross and/or histo evidence of negative energy balance.

Risk factors assessed for developing amyloidosis – clinical haemonchosis, pneumonia, negative energy balance, ruminal microabscesses.

Pedigree analysis demonstrated that 5 animals shared some degree of relatedness.

No statistically significant difference between mean degree of relatedness to each of the five founders for the animals with amyloidosis.

- Discussion:o Concluded that chronic inflammation secondary to haemonchosis and/or pneumonia

was the cause of systemic reactive amyloidosis in this herd (77%).o Some thought that fecal amyloid fibrils can transmit in a prion-like manner between

individuals. In cheetas, administered amyloid fibrils from fecal samples to mice and mice developed amyloidosis.

o Pronghorn antelope SAA and haptoglobin sera levels did not correlate with presence of amyloid histologically, lesions associated with negative energy balance or pneumonia, or clinical documentation of haemonchosis.

o Evaluation of pedigree did not show an obvious hereditary association.o There was a tendency for negative energy balance lesions, ruminal microabscesses,

pneumonia, statistically significant assoc with clinical haemonchosis and amyloidosis. Chronic inflammation with or without concurrent acute inflammation caused

systemic amyloidosis in this herd, leading to clinically significant renal disease.

Bartlett, S. L., Arheart, K. L., & Garner, M. M. (2019). Retrospective analysis of mortality in captive bongo (tragelaphus eurycerus), 1995–2015. Journal of Zoo and Wildlife Medicine, 50(2), 303-307.

Abstract: A retrospective histologic study was performed for 96 deceased bongo (Tragelaphus eurycerus) submitted to Northwest ZooPath from 1995 to 2015. Histologic data were assessed for associations with sex, age, cause of death/euthanasia, and affected organ systems. Female bongo lived significantly longer than males. Males were more likely to die from infectious causes (41.2%), whereas most females died from chronic noninfectious conditions (54.4%) and trauma/stress (28.1%). Of those that died from infectious disease, the respiratory tract was the most commonly affected organ system. The most common infectious agents included acid-fast bacteria and fungi. Chronic conditions included amyloidosis (31.0%), inanition/emaciation (23.8%), and neoplasia (21.4%). Of the 31 animals that died with amyloidosis, 58% appeared to be clinically affected, and amyloidosis was likely an underlying cause of death in 42% of the animals. The most commonly affected organs included the liver, kidneys, adrenal glands, and gastrointestinal tract. Also noteworthy was a high prevalence of adrenal gland hyperplasia and neoplasia, cystic thyroid glands, and aspiration pneumonia, which were not consistently associated with a prior anesthetic event or other obvious predisposing cause.

- Introduction:

o Bongo (Tragelaphus eurycerus) – forest-dwelling bovids found in central Africa. Two subspecies – mountain aka eastern bongo (T. e. isaaci), lowland aka

western bongo (T. e. eurycerus). Commonly displayed in zoos.

o Mortality has been attributed to mycobacterium infections, malignant catarrhal fever, amyloidosis, abomasal impaction, uterine leiomyoma, and osteofluorosis.

Placental tumor in stillborn fetus. Death due to complications assoc with failure of passive transfer. Familial goiter has been described.

o Aim – ID common causes of mortality in bongo at some US zoological facilities and to ID any associations with sex and age.

- M+M:o Retrospective histo study for 96 bongo submitted to NW zoo path from 1995-2015.

- Results:o 96 animals, 31 euthanized, 61 died, 4 not specified.

30 institutions represented. 34 males, 57 females, 5 unspecified. Females lived significantly longer than males.

o Most common causes of death were chronic dz (43.8%), trauma/stress (27.1%), and infectious disease (25%).

Females tended to die from chronic conditions (54.4%) and trauma/stress (28.1%). Males more likely to die from infectious causes (41.2%).

Adult and geriatric animals died with chronic conditions (44.1 and 88.9%, respectively).

Neonatal animals died from congenital/developmental problems, trauma/stress, infectious disease.

Most common cause of anesthetic death systemic response (shock with rhabdomyolysis).

For chronic causes of death – conditions affecting the urinary system, GI system, and systemic causes predominated.

Most common histo dx for urinary system – chronic interstitial nephritis, nephrosclerosis.

Chronic GI problems leading to death commonly secondary to amyloid deposition.

For trauma/stress, resp system most commonly affected, with 12 animals dying from aspiration pneumonia.

Respiratory system most commonly affected by infectious disease. Most common cause of pneumonia was infection with acid-fast

bacteria, presumed or confirmed to be Mycobacteria spp. Four animals euthanized due to exposure to animals that died with TB

lesions. Four animals died of systemic fungal infections including Mucorales, Cladosporium spp.

o Most common types of chronic conditions that contributed to cause of death included amyloid (31%), inanition/emaciation (23.8%), neoplasia (21.4%).

Although not always cause of death, neoplasia in 20 animals. Uterine neoplasia (leiomyomas), pheochrommocytomas, adrenal

medulla hyperplasia, adrenal cortical nodular hyperplasia, nerve sheath tumor in adrenal gland.

High prevalence of cystic thyroid glands.o Animals that died with amyloid significantly older.

No assoc between sex and amyloidosis. 32% animals had some degree of amyloid. 58% of those appeared clinically affected. Likely underlying cause of death in 42% animals.

Most severe in pancreatic duct, ovary, liver, omasum, adrenal glands and intestine.

- Discussion:o Bongo appear particularly susceptible to conditions affecting the respiratory tract.

Mycobacterium documented previously, acid-fast positive organisms found in this review. M. avium and M. kansasii identified in two cases.

Nocardia and rhodococcus equi other possible acid-fast staining organisms. 10% animals in this study had aspiration pneumonia. Occ assoc with anesthesia

or lumpy jaw, but also in adults with chronic weight loss with or without increased respiratory effort.

Cystic thyroid glands common (43/53 bongo), significance of lesion unknown.o Species predilection for amyloidosis.

Captive bighorn sheep (Ovis Canadensis) susceptible. Arabian gazelles (Gazella Arabica), all urinary diseases moderate to severe amyloidosis. Renal medullary amyloidosis with subsequent renal failure in a herd of Dorcas gazelles. Sand gazelle (Gazella leptoceros) and Speke’s gazelles (Gazella spekei) noted to die of systemic amyloidosis.

Amyloidosis in this study significant cause of mortality in 6.1%.o Amyloid in GIT appeared to be assoc with emaciation, malabsorption, dysbiosis,

gastrointestinal stasis and torsion. Amyloid in rumen could potentially predispose to aspiration.

o Strongly consider amyloidosis as a primary or secondary problem in bongo, particularly older animals.

HISTOPATHOLOGIC CHARACTERISTICS OF THYROID GLAND NEOPLASMS IN THOMSON'S GAZELLES (EUDORCUS THOMSONII)

French SJ, Garner MM, Kiupel M. Journal of Zoo and Wildlife Medicine. 2018 Mar;49(1):64-9.

Abstract: Published reports of neoplasms in Thomson’s gazelles (Eudorcas thomsonii) are very rare, but thyroid tumors were the most common neoplasm of this species, accounting for 12% of reported pathologies in a 1998–2012 retrospective study of cases submitted for histologic review of grossly enlarged thyroid glands. This report describes the histological and immunohistochemical characteristics of thyroid neoplasms in 10 Thomson’s gazelles from five different zoological collections. Neoplasms were submitted as biopsies from six gazelles or collected during necropsy from four gazelles. The most common clinical findings included a palpable mass on the ventral neck and progressive weight loss. Radiographic mineral density was detected in one of the neoplastic masses. Histologically, the neoplasms

were classified as microfollicular thyroid adenoma (n = 2), solid thyroid adenoma (n =2), papillary thyroid adenoma (n = 1), and solid thyroid carcinoma (n = 5). Neoplastic cells in all 10 neoplasms were positive for thyroid transcription factor 1 and thyroglobulin, but negative for calcitonin. While five cases had histologic features of malignancy, there was no evidence of metastatic disease either clinically (biopsies) or on necropsy. Numerous concurrent diseases, including cardiomyopathies and nephropathies, were present and led to choice for euthanasia in several cases. INTRODUCTION/METHODS

- Reports of neoplasms in Thomson’s gazelles rareo Thyroid tumors most common (12% of reported pathologies in a 1998–2012 study of 83

cases submitted for histologic review of grossly enlarged thyroid glands)- Report describes the histological and immunohistochemical characteristics of thyroid neoplasms

in 10 Thomson’s gazelles from five different zoological collections RESULTS/DISCUSSION

- Avg age w/ neoplasm = 12.8y; hx complaint: weight loss, palpable cervical mass- Microfollicular thyroid adenoma (n = 2), solid thyroid adenoma (n =2), papillary thyroid

adenoma (n =1), and solid thyroid carcinoma (n =5)- The most common clinical finding reported was a palpable SQ mass(es) on the ventral neck

o All positive for thyroid transcription factor 1 and thyroglobulin, negative for calcitonin Confirms thyroid follicular cell origin

o 5 malignant carcinomas, but no metastasis- Comorbidities (nephropathy, cardiomyopathy), rather than thyroid dz reason for euthanasia- Thyroid tumors of gazelles most analogous to disease in dogs

Take home: Thyroid neoplasms overrepresented in Thomson’s gazelles (suspect #1 neoplasm)

CAPTIVE MANAGEMENT OF WILD IMPALA (AEPYCEROS MELAMPUS) DURING INTENSIVE IMMOBILIZATION AND GENERAL ANESTHESIA STUDY TRIALS

Zeiler GE, Meyer LC. Journal of Zoo and Wildlife Medicine. 2017 Dec;48(4):1058-71.

Abstract: Immobilization and anesthesia of impala (Aepyceros melampus) has become a popular research theme. This demand is brought about by the increased need to immobilize and anesthetize impala and other medium-sized wild ungulates because of their increased value in game ranching and zoological collections. To improve our understanding of immobilization and general anesthesia in these species, it is paramount to be able to study them in a practical, safe research environment that does not cause harm or unnecessary stress to the animals. This study aimed to scrutinize our management and welfare practices and scientific methods of 15 wildcaught impala placed in captive confinement during a 16-wk intensive

research project. The scientific methods of the project were scrutinized to identify procedures that attributed to morbidity and mortality. Indicators of impala welfare during captivity were monitored by documenting serial physical (body weight, coat condition), physiological (biochemistry and hematology), and psychologic (behavioral) aspects. Two impala suffered irreparable femoral fractures during darting and were humanly euthanized. One impala suffered cardiovascular collapse during immobilization and could not be resuscitated. The procedure of chemical capture had a morbidity and mortality rate of 14.8% and 4.4%, respectively. The impala maintained acceptable physical and physiologic parameters, including stable body weights, well-groomed coats, and values for total serum protein, serum creatinine, and hematologic parameters that were within reference ranges for the species. There were improvements in the impalas’ psychological parameters, which included a decrease in the number of aggressive interactions (head butting and ramming) and an increase in the number of reciprocal allogrooming interactions. The monitored welfare indicators suggest adaptation to captivity. The study showed that impala could be successfully managed in captivity for 16 wk. However, scientific methods (namely darting) increased the risk of injury and caused fatalities.

Introduction/Methods: Impala are challenging in captivity. Prone to malasaption, anaorexia, injuries, myopathy Research needed into management and welfare to ID risks and appropriate practices Factors to be evaluated:

Physical state (BCS, wt, coat condition, lameness, presence of diarrhea, etc.) Physiologic state (biochemistry, hematology, cortisol) Psychological state (behavioral changes)

Evaluated welfare risks and outcomes for a group of wild-caught impala used in an anesthesia protocol research

All impala captured 6 times through 16wks of the study. Were kept in a boma with separated ‘home’ and ‘feeding’ areas

Impala initially administered zuclopenthixol during move from origin to boma Boma size exceeded recommended size for short term housing in S. Africa

Study determined rate of mortality and morbidity and risk factors Study collected serial data on physical, physiologic and psychologic welfare

Results/Discussion Boma and husbandry techniques caused no adverse effects. Experiment procedures (darting) led to 3 deaths – 1 cardiac arrest, 2 femur fractures

10 additional non-fatal adverse events (injuries/cardiovascular derangements) during darting Morbidity – 14.8%, Mortality - 4.4% Closer darting or a lower pressure of darts may have reduced adverse events

No physiologic derangements – All CBCs and Chem panels were within RI Cortisol remained unchanged over 16 wk period

Antagonistic interactions decreased in frequency over 16week period Individual and reciprocal allogrooming noted from week 9 onwards in the herd. Developed decreased reaction to investigators around week 9.

Takeaways: Results suggest adequate adaptation by the impala to the boma and husbandry practices. Characteristics of a successful Boma described by authors as:

Having adequate size for the number of animals housed (boma here was larger than S. African ‘Norms and Standards’ but smaller than zoo paddock recommendations)

Having space for a latrine separate from feeding, drinking, and resting areas Provision of shelter Internal partitions with high wall to allow sense of escape for impala when humans enter Interleading swing gates allowing confinement of Impala to one part of the enclosure High walls to prevent escaper and injury through jumping

Ample gates to allow access to both boma partitions from outside Allowance of adequate ventilation

Current capture methods for impala from boma led to significant adverse effects (fatalities and injuries from darting events)

DISTAL LIMB SWELLING AND PERIOSTEAL PRODUCTIVE REACTION IN PERIPARTURIENT SICHUAN TAKIN (BUDORCAS TAXICOLOR TIBETANA): FIVE CASES OF PRESUMPTIVE HYPERTROPHIC OSTEOPATHY

Journal of Zoo and Wildlife Medicine 50(2): 437–446, 2019

Marinkovich, Matt, Wisner, Erik R., and Brenner, Deena J.

Abstract: Hypertrophic osteopathy, a syndrome characterized by painful distal limb swelling and proliferative periosteal reaction, primarily involves the metacarpal and metatarsal bones and phalanges. Lesions are often bilaterally symmetric and typically affect all four limbs. Hypertrophic osteopathy is frequently associated with primary intrathoracic disease, though this condition has also been reported secondary to intra-abdominal and intrapelvic disease and associated with pregnancy in both people and horses. Over a 20-yr period, five adult female Sichuan takin (Budorcas taxicolor tibetana), with 13 total pregnancies, were evaluated because of lameness and distal limb swelling. These clinical signs were observed between 2 and 32 days (mean ¼ 19 days) prior to parturition (gestation period in takin approximately 200–240 days) and resolved in all animals following parturition. Lameness and limb swelling resolved between 8 and 168 days (median¼15 days) after parturition. Sixteen radiographic examinations, from four of the individuals, documented proliferative periosteal reaction, primarily of the metacarpal bones, metatarsal bones, and phalanges. The clinical progression, resolution of signs, and radiographic features in these cases are consistent with hypertrophic osteopathy, secondary to pregnancy. This is the first report describing presumptive hypertrophic osteopathy in takin.

Summary/key points:

Hypertrophic osteopathy (HO) - syndrome characterized by firm, often painful, slowly progressive soft tissue swelling of distal limbs that seldom affects articular or synovial tissues

o Clinical signs - lameness, stiff gait, reduced flexibility of distal limb joints, pyrexia lethargy

o Rads - proliferative periosteal reaction along diaphysiso metacarpal and metatarsal bones and phalanges, proximal long bones less commonly

involved o lesions often bilaterally symmetric and affect all four limbso frequently associated with primary intrathoracic diseaseo diagnosis – combination of rads, clinical signs, and identification of a primary lesion

(either intra- or extrathoracic) Case series - 5 adult female Sichuan takin diagnosed with presumptive HO secondary to

pregnancyo clinical signs ultimately resolved following parturition in all caseso elevated WBC count along with hyperfibrinogenemia was seen in 4 of 5 caseso no difference in calf size, gestation length, number of parturitions, and time between

pregnancies between dams with and without HO

vascular endothelial growth factor (VEGF) expression may be the result of 3 potential pathophys mechanisms: abnormal vascularization, hypoxia, and chronic inflammation

o VEGF plays a role in development and progression of pathophys changes associated with HO

o Pregnancy creates a proinflammatory environment

VALIDATION OF IMMUNOCRIT AS A DIAGNOSTIC TEST TO DETECT FAILURE OF PASSIVE TRANSFER OF IMMUNITY IN HAND-REARED NONDOMESTIC RUMINANT NEONATES

Journal of Zoo and Wildlife Medicine 50(3): 627–633, 2019

Thompson, Kimberly A., Lamberski, Nadine, Rayburn, Maire, and Chigerwe, Munashe

Abstract: Domestic and nondomestic ruminant neonates with failure of passive transfer of immunity (FPI) have an increased risk of morbidity and mortality. Hand-reared individuals often have a higher incidence of FPI compared with those that are dam-reared. Hand-reared, nondomestic ruminant neonates raised at zoological establishments are often fed a commercially available bovine colostrum replacer. Consequently, it is desirable to monitor for FPI using a single test that performs well across a wide range of species. The immunocrit test method has been reported to have acceptable diagnostic sensitivity for detecting FPI in dairy calves. Advantages of the immunocrit are that it is inexpensive, quick, easy to perform, non-species-specific, and only precipitates immunoglobulins. This study evaluated the ability of immunocrit to predict serum immunoglobulin levels and FPI via comparison with radial immunodiffusion (reference method). The study used 100 serum samples collected from hand-reared ruminant neonates from 29 nondomestic species, 12–120 hr old, that were fed a commercial bovine colostrum replacer at a zoological institution. Sensitivity, specificity, and likelihood ratios for FPI were determined using receiver operating characteristic (ROC) curves. The immunocrit cutoff point of 14 % maximized sensitivity at 88.6 % (95 % CI, 78.7–94.9 % ) and specificity at 83.3 % (95 % CI, 65.3–94.4 % ). The area under the ROC curve was 0.92 (95 % CI, 0.87–0.98; P , 0.001). This research demonstrates that the immunocrit test has acceptable performance in detecting FPI in a wide variety of nondomestic hand-reared ruminant neonates. Use of the immunocrit test will help improve the identification of FPI neonates and aid in the management of colostrum feeding practices of nondomestic ruminant species in captivity.

Summary/key point:

Failure of passive transfer of immunity (FPI)o negligible transfer of antibodies across ruminant cotyledonary placentao neonates rely on ingestion of colostrum and absorption of maternal antibodies across

the intestines during first 24–48 hr after birtho IgG measured to assess passive transfer

radial immunodiffusion (RID) or enzyme-linked immunosorbent assay (ELISA) used for direct measurement

passive transfer of immunity cutoff point of serum IgG levels is 1,000 mg/dl immunocrit method - precipitation of immunoglobulins with a known concentration of

ammonium sulfate solutiono not species-specifico results are semiquantitativeo can be performed inhouse quickly, affordably, and with minimal equipment and training

o this study found good accuracy in identifying those samples with FPIo Springbok found to have poor accuracy of only 50%

immunocrit recommendations may vary with species Radial immunodiffusion considered a species-specific reference test while the precipitation of

immunoglobulins by immunocrit has been shown to be non-species-specific

RETROSPECTIVE EVALUATION OF HISTOPATHOLOGIC FINDINGS IN CAPTIVE GAZELLE SPECIES

Kadie Anderson, Michael Garner, Nancy Stedman

J Zoo Wildl Med, 47 (1), 113-9 Mar 2016

Taxa: Mammalia → Artiodactyla → Bovidae → Antilopinae

Abstract: Capturing disease trends among different species has indisputable value to both veterinary clinicians and zoo managers for improving the welfare and management of zoo species. The causes of mortality for eight species of gazelle (addra gazelle, Nanger dama; dorcas gazelle, Gazella dorcas; Grant's gazelle, Nanger granti; sand gazelle, Gazella leptoceros; Saudi goitered gazelle, Gazella subgutturosa; Soemmerring's gazelle, Nanger soemmerringii; Thomson's gazelle, Eudorcas thomsonii; and Speke's gazelle, Gazella spekei) are presented from an 18-yr period (1996 2014). The leading cause of mortality for all species was trauma, followed by bronchopneumonia, and failure to thrive/maternal neglect. Nephritis and rumenitis/abomasitis/enteritis were common ancillary lesions across all species. On average, female gazelle lived twice as long as male gazelle, with an average overall adult survival

time of 9.3 yr. Dorcas, Thomson's and addra gazelle females had the longest average survival time (10-13 yr). Calves up to 6 mo of age died most frequently from failure of passive transfer or maternal neglect. Thyroid carcinoma was frequently identified in Thomson's gazelle. Sand and Speke's gazelle frequently died from systemic amyloidosis, and Saudi goitered gazelle were more likely to have renal amyloidosis. Hematuria syndrome was the second most common cause of death in Grant's gazelle. The majority of lesions identified in this study that cause or contribute to mortality are preventable with appropriate management. Knowledge of disease trends in captive gazelle populations can help guide veterinary care, management decisions, and collection management planning.

Methods: Retrospective analysis of histopathologic lesions in gazellees submitted to Northwest ZooPath (1996-2018)

Key Points:

- Overall, females lived twice as long as their male conspecifics- Causes of adult death: trauma (32.5%) >>>> bronchopneumonia (11.3%) >> maternal neglect

(8.4%) > other- Bronchopneumonia was commonly associated with an anesthetic event (45.4%)- Young calves commonly died from maternal neglect, failure of passive transfer and sepsis- Older calves commonly died of trauma- Rumenitis / enteritis was a common secondary finding (7.7%)- Thomson’s gazelles had a relatively high incidence of thyroid neoplasia (adenocarcinoma,

carcinoma, and adenoma)- Grant’s gazelles had a high incidence of hematuria syndrome with no identifiable etiologic agent

- Hematuria syndrome = hematuria without pyuria, submandibular edema, hypocalcemia, hypoproteinemia, hypoalbuminemia, hyperphosphatemia, +/- anemia

- Amyloidosis was relatively common (6.1%) in sand, Saudi, Soemmering’s, Grant’s, and Speke’s gazelle, presumed secondary to chronic inflammation

- National Zoo study in dorca’s gazelles found association with chronic Arcanobacterium infection

Conclusions: Captive gazelles most commonly die from trauma, though many other causes of death occur.

THE EFFECT OF A SLOW-RELEASE FORMULATION OF ZUCLOPENTHIXOL ACETATE (ACUNIL®) ON CAPTIVE BLUE WILDEBEEST (CONNOCHAETES TAURINUS) BEHAVIOR AND PHYSIOLOGICAL RESPONSE

Liesel L Laubscher, Louwrens C Hoffman, Neville I Pitts, Jacobus P Raath

J Zoo Wildl Med, 47 (2), 514-22 Jun 2016

Taxa: Mammalia → Artiodactyla → Bovidae →

Abstract: The study investigated the effect of a slow-release formulation of zuclopenthixol acetate (Acunil®) on blue wildebeest (Connochaetes taurinus) in captivity. Two groups of trials were conducted using either Acunil or a placebo (control). Animals (Acunil: n = 17; placebo: n = 12) were observed for a 12-hr period before the administration of Acunil or the placebo (pretreatment). After 24 hr, animals were administered Acunil (1.5 mg/kg) or a placebo (1.0-3.0 ml of sterile water) and observed again for 12 hr (posttreatment). During both treatments, animals were stimulated every 2 hr for 1 min by a person entering the enclosure (referred to as periods of stimulation). Behavioral observations and continuous heart rate, respiration rate, and motion measurements were taken throughout. Animals treated with Acunil spent more time lying with their heads folded back, eating and standing with their heads down, and less time being vigilant and exploring while walking around. Animals treated with the placebo also spent less time being vigilant and more time lying with heads up. Animals treated with Acunil groomed less while standing and performed less head shaking; no such changes were observed in the control group. Neither Acunil nor the placebo had any effect (P > 0.05) on heart rate. However, overall mean respiration rate was lowered (P = 0.02) when animals were treated with Acunil (pretreatment: 14.5 ± 0.82 breaths/min; posttreatment: 12.5 ± 0.83 breaths/min). Acunil also caused a lowered (P < 0.05) respiration rate during periods when animals were stimulated (pretreatment: 16.2 ± 0.87 breaths/min; posttreatment: 13.7 ± 0.87 breaths/min) and when animals were trotting and being vigilant. No such changes were observed with the placebo. Both placebo- and Acunil-treated animals spent more time being stationary during periods of stimulation. However, Acunil-treated animals also spent less time moving fast when they were stimulated.

Background:

- Long-acting neuroleptics (LAN): dopamine-blocking antipsychotics dissolved in oil to form a depot at injection site

- Clopixol-Acuphase: zuclopenthixol acetate dissolved in vegetable oil- Acunil: zuclopenthixol acetate dissolved in 72-hour slow-release polymer (new WildPharm

product)

Methods: Wildebeest vial signs, movement, and behaviors observed for 12 hours before and after treatment with Acunil 1.5 mg/kg or placebo IM.

Key Points:

- Acunil likely decreased stress in wildebeest because:- Lower respiratory rate (including when stationary, trotting, and vigilant)- Less grooming when standing (grooming = stress displacement behavior)- Less head-shaking- More time stationary- Less time moving quickly following stimulation- More time laying with head turned back- More time eating

- Control group also had more time stationary and less time moving slowly, likely from habituation

- No change in heart rate - No extrapyramidal signs nor adverse effects

Conclusions: Acunil, a new zuclopenthixol acetate formulation, was safe and was effective in decreasing stress in wildebeest.

BILIRUBIN CONCENTRATIONS IN CLINICALLY HEALTHY AND DISEASED CAPTIVE WATERBUCK (KOBUS ELLIPSIPRYMNUS) AT THE SAN DIEGO ZOO SAFARI PARK

Ryan A Sadler, Nadine Lamberski, Mary M Christopher

J Zoo Wildl Med, 47 (2), 523-30 Jun 2016

Taxa: Mammalia → Artiodactyla → Bovidae → Reducinae (subfamily)

Abstract: Captive waterbuck (Kobus ellipsiprymnus) that appear clinically healthy have been noted to have high serum bilirubin concentrations compared with other ruminants; however, questions remain about the physiologic factors affecting bilirubin concentration and its potential association with underlying disease and icteric serum or mucous membranes. Serum bilirubin concentrations of healthy and diseased waterbuck housed at the San Diego Zoo Safari Park from 1989 to 2012 were retrospectively analyzed to determine any link between icteric serum, total bilirubin concentration (tBili), and disease entities in this species. Total bilirubin and direct (dBili) bilirubin concentrations and the prevalence of icteric serum were compared by subspecies, age group, and health status; associations with complete blood count and biochemical results and clinical diagnosis were assessed. No significant differences were found in tBili or dBili between Ellipsen (n = 32) and Defassa (n = 29) subspecies or in juveniles (n = 22) versus adults (n = 39). Clinically healthy waterbuck (n = 40) had significantly higher tBili (mean ± 2SD, 7.9 ± 1.2 mg/dl; P < 0.001) and dBili (3.7 ± 1.0 mg/dl; P < 0.001) than did diseased waterbuck (n = 21; tBili: 4.9 ± 2.56 mg/dl; dBili: 2.2 ± 0.8 mg/dl). No waterbuck had icteric tissues on physical examination. Twelve (19.7%) waterbuck (six healthy, six diseased) had icteric serum. Few minor correlations were seen between tBili or dBili and clinical, laboratory, or necropsy evidence of disease, though an inverse correlation between dBili and blood glucose was noted. Of the 40 healthy animals, reference intervals were calculated for tBili (5.5-10.3 mg/dl), dBili (1.7-5.7 mg/dl), and indirect bilirubin (2.2-6.2 mg/dl). These results suggest healthy waterbuck have relatively high tBili and dBili compared with related species. Icteric serum may be seen in up to 15% of healthy animals in the absence of icteric tissues.

Background:

- Bilirubin: product of heme breakdown that is conjugated in the liver, then excreted in bile- Indirect bilirubin (iBili) = unconjugated bilirubin- Direct bilirubin (dBili) = conjugated bilirubin- Total bilirubin (tBili)) = iBili + dBili

- Differentials for increased bilirubin:- Increased hemolysis- Impared liver function- Cholestasis- Fasting (in horses and some ruminants; lipolysis interferes with bilirubin uptake from

hepatocytes, increasing unconjugated bilirubin)- Waterbuck are known to have very high tBili and dBili

Methods: Retrospective review of tBili and dBili in waterbuck at the San Diego Zoo Safari Park (n=61, 1989-2012).

Key Points:

- Healthy waterbuck had higher tBili and dBili than diseased waterbuck- High bilirubin levels did not correspond to icteric serum (15% of samples) nor icteric mucous

membranes (0% of animals)- Healthy waterbuck had higher albumin, glucose, and PCV, and lower total WBC counts- In healthy waterbuck, glucose and dBili were inversely related.- Diseased waterbuck had higher creatinine kinase (two individuals had capture myopathy)- Disease waterbuck with high tBili (>3) had lower albumin than diseased waterbuck with lower

tBili- Myoglobin contains one heme molecule, and may lead to hyperbilirubinemia in severe

myopathy cases- No differences between subspecies and age (between juveniles and adults)- Reference intervals were calculated (tBili = 5.5-10.3 g/dL; dBili = 1.7-5.7)- All neonates evaluated were diseased and had lower tBili than juveniles and adults

Conclusions: Because healthy waterbuck have higher bilirubin concentrations than diseased waterbuck, serum bilirubin concentrations are not an accurate indicator of liver dysfunction in this species.