Felidae Flashcards

Question

What herpesvirus affects captive cheetahs? How is it transmitted? What are the clinical signs? What demographics are more suceptible? Can it be induced with modified live vaccine?

Answer 1

Journal of Zoo and Wildlife Medicine 51(1): 210–216, 2020 **FELINE HERPESVIRUS INFECTION IN FOUR CAPTIVE CHEETAHS (ACINONYX JUBATUS) POSTVACCINATION** Alexis Pennings, DVM, Kathryn Seeley, DVM, Dipl ACZM, MS, Ame´ lie Mathieu, DMV, MS, Abby Foust, DVM, Dipl ACVD, Michael M. Garner, DVM, Dipl ACVP, and Jan Ramer, DVM, Dipl ACZM. Abstract: Cheetahs (Acinonyx jubatus) are particularly susceptible to feline herpesvirus-1 (FHV-1). Recommendations for preventive health care in cheetahs include vaccination against FHV-1 using killed and modified live virus (MLV) vaccines. Although MLV vaccines tend to induce a more robust immune response than killed vaccines, they can induce disease. **This case series details an FHV-1 outbreak in four adult cheetahs following the use of MLV vaccine in one of them. All four cheetahs developed severe FHV-1 clinical signs and were euthanized.** Clinical signs included **depression, anorexia, nasal discharge, ocular discharge, sneezing, and ulcerative dermatitis.** Herpesvirus infection was diagnosed using history, clinical signs, polymerase chain reaction, and histologic evaluation. The timeline of events suggests the MLV vaccine was the inciting cause, although this was not conclusively proven. Outcome of this case suggests that when considering MLV vaccines for cheetahs, careful risk and benefit discussions are merited. Key Points: * Feline herpesvirus-1 – alphaherpes virus, endemic in managed cheetah, also found in semi-managed and free-range * Transmission between cheetah or other felids * Inactivated in environment within hours * Oronasal entry -\> clinical signs within 2-6d * Latency and recrudescence – viral reactivation common after stressful event, takes about 4-11 days before shedding, may last for 13 days * Clinical signs in cheetah – depression, inappetence, conjunctivitis, corneal ulceration, nasal discharge, sneezing, ulcerative dermatitis * Young cheetah more likely to have severe disease +/- death * MLV administration prior to developing disease has been reported multiple times in cheetah * Cheetah at the institution were historically vaccinated with killed vx; however – brought in female for breeding and based off program recommendations she received a killed booster then a MLV 20d later * Approx 1 month after MLV administered to female – first male became ill – pos on nasal swab for FHV-1; other two males then became ill. All 3 males euthanized because of deteriorated health and elderly * Female also broke with disease – attempted therapy for 15 months then euthanized * Two other females at the facility were also vaccinated with the same protocol at the same time and did not develop disease * Cell mediated immunity plays an important role for fighting FHV-1 so titers not the best way to monitor Take home: Using MLV for viruses in exotic felids is not the best idea.

Answer 2

**BIOLOGY (Fowler 8):** * The family Felidae consists of at least 36 wild cat species * Number of genera has Been variable but 12 genera currently recognized * Felis (black footed cat, sand cat) * Otocolobus (Pallas) * Lynx (lynx, bobcats) * Caracal * Leptailurus (serval) * Pardofelis * Prionailurus (Fishing cat) * Leopardus (Ocelot) * Puma (Puma, Jaguarundi) * Neofelis (Clouded leopard) * Panthera (Snow leopard, tiger, leopard, jaguar, lion) * Acinocyx (cheetah) * Occupy diverse habitat, distributed throughout the world (except Antarctica and Australia) * Among the most threatened groups of mammals (danger to humans/livestock, fur trade) * Nearly 50% of all felids in top 3 threatened categories, 7 Critically Endangered or Endangered. * Iberian lynx (Critically Endangered), may be first cat species to become extinct in modern times

Answer 3

**ANATOMY (Fowler 8):** * 28-30 teeth, formula: incisors (I) 3/3, canines (C) 1/1, premolars (P) 2-3/2, molars (M) 1/1. * Length wise groove in canines * The fourth upper premolars (carnassial teeth) are used to slice meat * More powerful bite relative to muscle mass than any other carnivore (except mustelids) * Due to carnivorous diet, have shorter digestive tract, smaller cecum, short large intestine. * Forelimbs used for locomotion and prey capture- need supination of paw. * Increased mobility of the elbow and wrist joints affects running. * Canids run faster as they have stiffer forelimbs. * The hindlimbs are propulsers, and reduction or loss of a clavicle increases stride length. * Cats are digitigrade, with five toes in the front and four in the back. * First digit on front foot is the dewclaw. * All felids have retractable claws * _Exception = cheetah_. Cheetahs lack the fleshy sheath protecting the claw * Previous thought: Key characteristic to separate big cats (Pantherinae) from small (Felinae) = * Presence of an elastic ligament in the hyoid apparatus below the tongue * Thought to allow the big cats to roar but not purr. * Conversely, bony hyoid of small cats was thought to allow them to purr but not roar. * New thought: Difference btwn roaring, nonpurring cats = long, fleshy, elastic vocal folds (larynx) * Found in big cats that resonate to produce a roar. * S_maller cats_ and _cheetahs_ have simpler vocal folds that only allow purring * Nondomestic felids have AB blood group system similar to domestic cats. * Cross-matching of donor and recipients important before transfusions **Unique Features (Terio Ch 10)** * Dental formula as domestic felids (13/3, C1/1, PM2-3/2, M1/1 in each arcade) * Zonary placentation with endotheliochorial maternal-fetal interface * Clouded leopards have unusually long canine teeth * Cheetah claws are semi-retractable, metacarpal and metatarsal pads are rigid * Large hearts in proportion to body size * Ectatic pancreatic ducts – variably sized, clear, fluids filled cystic structures within the pancreatic parenchyma * Black footed cats have enlarged adrenal glands * Prominent esophageal folds in cheetahs and some *Panthera* (stratified squamous) * Wolffian duct (mesonephric duct) – embryonal paired structure -\> epididymis, vas deferens, and seminal vesicles in males * In females, ducts regress in absence of anti-mullerian hormone but cysts can develop in remnant ducts * Mesonephric duct cysts in all felids; paraovarian cyst in female cheetahs common * No association between presence of cysts and fertility * Hepatic telangiectasia – uncommon – cheetahs, snow leopards, cougars, & lions * Sinusoidal dilation

Answer 4

**SPECIAL HOUSING REQUIREMENTS (Fowler 8):** * To reduce osteoarthritis and pad ulceration, do not house on concrete for long periods * Appropriate safety precautions must be designed into the enclosure and holding * Safety gates provide secondary containment if an animal escapes from the primary holding * Keepers may carry pepper spray and communication radios while working with large felids. * Escape drills should be held routinely. **FEEDING (Fowler 8):** * -Require much higher protein in their diet compared with any other mammal * -12% for domestic cats compared with 4% for domestic dogs. * -Most captive felids fed commercial meat-based complete diets * -Additional vitamin/mineral supplements (calcium/taurine) should not be necessary * -Techniques for proper thawing of frozen diets via https://www.aphis.usda.gov/animal\_welfare/downloads/big\_cat/handlemeat.pdf * Freezer: –30 to –18 o C (–22 to 0 o F) or lower * Refrigerator: 4 to 6 o C (40 to 43 o F) * Once removed from the freezer for thawing or thawed under refrigeration, meat must be used within the next 24 hours. * -Bones may be offered to promote oral health and to provide enrichment * -Diet should be analyzed for nutritional content, bacterial contamination, foreign material. * -Several cases of ethylene glycol toxicity from contaminated meat reported * -Source of diet should be checked to prevent spread of dz (spongiform encephalopathies)

Answer 5

**PREVENTATIVE MEDICINE (Fowler 8):** _Routine Health Examination:_ * -Many institutions perform examinations under anesthesia every 2 to 4 years * -freq. depends on animal’s age, life stage, health status and medical history, species * -Routine exam should include: BCS, body weight, PE, eval for ectoparasites, blood collection * -Recommended serologic tests: * -feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) * -Additional tests that may be necessary based on species, geographic location, or potential for disease exposure include those for * -*Toxoplasma gondii,* feline coronavirus (FCoV), canine heartworm (*Dirofilaria immitis*)*.* * *-*Serologic tests to monitor response to vaccination: * -feline parvovirus (FPV) esp. in _cheetahs_ * -rabies * -Vaccine titers for feline herpesvirus (FHV) or feline calicivirus (FCV) are predictive of protection, * -except for highly susceptible species such as _cheetahs_. * -If cheetah has low or negative titer to FPV or FHV, frequent vaccination should be considered. _Vaccination_: * -Vaccines recommended are divided into: * -Core vaccines (recommended for all felids) * -Noncore vaccines (optional, depending on disease risk, not generally recommended) * -Vaccine-associated sarcomas rarely reported in nondomestic felids. * -Specific information on length of protection from vaccination is lacking. * -Recommendations for vaccination freq. cannot be made * -Most institutions vaccinate adults every 1 to 3 years using the core vaccines. * -Core vaccines include: * -Rabies (killed, e.g., Imrab 3 or recombinant canarypox-vectored) * -Feline panleukopenia, calicivirus, herpesvirus (killed, e.g., Fel-O-Vax PCT Plus). * -Noncore vaccines: * -Should be considered only in species _at risk_ * -Canine distemper virus (CDV) (recombinant canarypox-vectored), FeLV (killed). _Pre-shipment Evaluation and Quarantine_: * -Quarantine should occur in an off-exhibit area away from other animals. Usually 30 days. * -Dedicated tools and equipment and PPE should be used, footbath.

Answer 6

**ANESTHESIA (Fowler 8):** * Smaller species require a higher dosage of anesthetics compared with larger species * Drug combinations: dissociative, α2-agonist, benzodiazepine, opioid, or a combination of these. * Telazol may be used safely in felids but should be _used with caution in tigers_. * Adverse reactions: death, neurologic dz (seizures, ataxia) anecdotally reported * Regurgitation/vomiting during induction/recovery may occur with α2-agonists * Recommend fasting for adult felids 12-24 hrs and water several hours prior to anesthesia * Venipuncture similar to domestic felids: medial/lateral saphenous, jugular, cephalic, femoral * Lateral tail veins (larger felids) located at 2 o’clock and 10 o’clock positions. **_Heard/West Chapter 45: Felidae Anesthesia_** **Anatomy/Physiology** * Small felids \<20kg, large \>20kg * Tigers have the most anesthetic complications * Telazol - sudden death (Wack 2003) - debatable? * Seizures, apneic episodes **Physical Restraint** * Hand injection if docile, juvenile * Squeeze cage/kennel/traps * Syringe pole * Remote darting - 3-5mL, hind limbs or shoulder **Manual restraint** * \<16kg, nets, gloves * Hand inject * Induction chamber **Behavior Training - ideal** **Oral Sedation** * Pre-med to decrease stress of darting * Not predictable **Injectable Agents** * Ketamine (not alone) * 11-44mg/kg; muscle rigidity, salivation, seizures, rough recoveries. Use w/ caution in renal failure cats bc excreted unchanged by kidneys * Useful supplementation and maintenance (0.2-2mg/kg IM or IV) * For south chinese tiges and bobcats- females require more ketamine * Ketamine + Medetomidine * Reliable sedation in most large felids * Smooth induction 5-15 min, good muscle relaxation * Lasts 45 minutes w/o gas * Good depth, low volume, analgesia, reversible partly * Disadvantage: vomiting during induction, CV changes, venoconstriction, low pulse ox readings, hypertension, bradycardia, bradyarrhythmias, decreased CO, mild to moderate respiratory depression, increased urinary output, sensitive to noise * Caution in heart disease cats * Ketamine 3mg/kg + Medetomidine 0.03mg/kg IM for large cats * Ketamine 2.5-4.5mg/kg + Medetomidine 0.05 -0.07mg/kg * Ketamine + Xylazine * Used more before medetomidine was available * Similar anesthesia and side effects as above * Ketamine 3-10 mg/kg and Xylazine 0.3-1.0 mg/kg * Alpha 2 + midazolam first. Then ketamine after sedated. Decrease likelihood of seizures. * Supplementation: Alpha 2 should not be used as supplement in prolonged procedures. Ketmaine better. Propofol useful to increase time of immobilization but has to be IV. Prolonged procedures should have intubation and gas anesthesia * Benzodiazepine (midazolam, diazepam) or butorphanol added to alpha 2 and ketamine combos * Ketamine + Midazolam * Small felids or debilitated large cats * Smooth anesthesia, good CV function * Diazepam - IV only * Ketamine 5-10mg/kg + Midazolam 0.1 - 0.3mg/kg * Butorphanol 0.1-0.4mg/kg or oxymorphone at 0.03 - 0.05mg/kg * Ketamine + Midazolam + Butorphanol * Torb provides additional sedation and analgesia * Low total volume - good for darts * Healthy small felids; debilitated, very young or geriatric large felids * Telazol * Short induction and reliable plane of anesthesia * High margin of safety and few CV effects * Recoveries prolonged and stormy * Pain at injection site, only partially reversible * Tigers: neurological conditions (seizures, ataxia) and death (2-4 days after anesthetic event) * 1.6-4.2mg/kg large cats; 11mg/kg small cats * Small volume * Can combine with ketamine to have better recoveries (decreased telazol) * Medetomidine + Butorphanol + Midazolam * Smooth, fast induction, adequate plane for 40 minutes (then sudden recoveries) * Cheetahs - 0.035mg/kg medetomidine + 0.15mg/kg midazolam and 0.2mg/kg butorphanol * Full reversible! Good for animals with kidney or liver disease * Hypertension, bradyarrhythmias (improve by reversing atipamezole early on) * Althesin * Saffan = alphaxalone and alphadolone acetate * Must be given IV, contains cremophor * Wide safety margin, little cumulative effect * Fatal anaphylactic reactions * Not used in U.S. but used in South Africa * Cheetahs 6-8ml/cat * Adequate for minor procedures * Propofol * IV, bolus (0.2-2mg/kg IV) or CRI (0.02-0,2mg/kg/min); one bolus q 5-10min * Good for animals with liver or kidney disease instead of ketamine **Isoflurane/sevoflurane/halothane** * Since already sedated, low levels needed (0.5-1% for iso, 0.5-2% for sevo) **Antagonist**s * Alpha-2: Atipamezole, tolazoline, yohimbine * Medetomidine (1mg): atipamezole (3-5mg); or 0.1-0.45mg/kg IM * High alpha 2 selectivity * Caution in neuro patients because atipamezole can raise ICP * Do not use yohimbine for medetomidine - not reliable (resedation, ataxia) * Use IM * Xylazine : Yohimbine (0.04 - 0.3mg/kg IM or slow IV) or Tolazoline (0.5-2mg/kg IM or IV) * Mixed alpha 1 and alpha 2 specificity * CNS excitement, salivation, muscle tremors * Don’t use atipamezole - excitement * Opioid: Naloxone, Naltrexone, Nalmefene * Naloxone (0.002-0.04mg/kg IM/IV) = pure mu antagonist * Naltrexone (0.05 - 0.25mg/kg IM/IV) = mu, delta, kappa antagonist; longer ating; can be given PO * Nalmefene (0.03mg/kg IV) = mu antagonist * Benzodiazepines: Flumazenil and Sarmazenil * Flumazenil (0.01-0/2mg/kg or 0.008 mg/ mg of diazepam or midazolam) * High cost * Does not improve recoveries in large felids; benzos benefit smooth revoery * Very short half life in cats **Intubation** * Small cats - 4-10mm * Large cats 16-24 * Cougars 10-14mm * Supplemental breaths if ETCO2 \>55mmHg or PaCO2 \> 60mmHg **Complications** * Vomiting * Alpha 2s * Hypertension and bradycardia * Alpha 2s * Reverse, don’t use atropine * Arrhythmias * Bradycardia * \<100bpm in cats \<7kg * \<80bpm in cats weight 10-20kg * 40-60bpm in large felids * Caused by deep anesthesia, hypothermia, cardiac issues * Lighten the anesthesia, warm the patient * Glycopyrrolate (0.005-0.01 mg/kg IV or IM) * Atropine (0.01-0.04mg/kg IV or IM) * Only treat if its due to non-medetomidine related causes * Tachycardia * Inadequate depth, pain, hypotension from hypovolemia * Hypotension * Fluid bolus 15-20ml/kg over 15 min, then 10-20ml/kg/hr * Dopamine, dobutamine * Cardiac Arrest * Hypoventilation * Hyperventilation (panting) * Respiratory arrest - lighten anesthesia, position in sternal, GV26, doxopram? * Seizures * Seen more in tigers with high dose of ketamine or telazol * Treat with 0.1-0.5mg/kg IV diazepam/midazolam; or propofol 0.2-1mg/kg IV * Electrolyte imbalances * Hyperkalemia * Tigers, cougars, cheetahs * Measure q15-30 min * 20% of animals * Metabolic acidosis * Treat if \>6mmol. Insulin in dextrose (0.1-1unit/kg in 500ml 5% dextrose) * Calcium gluconate - protects heart from arrhythmias * Reverse alpha 2s **Sedation** * Post op, decrease activity, decrease anxiety * Benzos orally (0.15-0.5mg/kg); also stimulate appetite * Long term PO at higher doses - hepatic failure * Discontinue if lethargic, vomiting, anorexic * Acetylpromazine 0.03-0.1mg/kg IM or 0.1-0.2 mg/kg PO -- mild to moderate sedation for up to 6-8 hrs in cheetahs * Lower doses to ‘take edge off’ in aggressive animals without causing sedation * Azaperone 3mg/kg to smooth telazol recoveries in lions * Perphenazine enanthate 3mg/kg IM for long lasting sedation up to 2 wks without side effects in cheetahs * Zuclopenthixol acetate not recommended in cheetahs - severe side effects (inappetence, ataxia, TEL prolapse, extrapyramidal reactions). Don’t use in any cats. **Pain Management** * Opioids * Side effects - salivation, mydriasis, hyperthermia * Buprenorphine used a lot in cats (partial u agonist) - shown to be superior to other opioids. Few side effects, long duration of action, oral/buccal absorption * 6hrs * 0.01-0.02mg/kg IV/IM/PO * Oxymorphone and hydromorphone (0.05-0.1mg/kg IV or IM) * Oxy less side effects than hydro * Fentanyl: short acting. Bolus 2-5mcg/kg + CRI 0.1-0.3mcg/kg/min. * Butorphanol (k receptor): only for mild pain, short duration. Use in combo with other drugs like nsaids or local blocks. Ceiling effect at 0.4mg/kg * Tramadol/fentanyl patches * NSAIDs * Up to 24 hrs of analgesia. Slow metabolism in cats due to lack of glucuronidase. * Side effects of GI ulceration, nephropathy, impaired coagulation. * Meloxicam (0.2-0.3mg/kg SC/IV pre op then 0.1mg/kg SC or PO for 3 days), carprofen, ketoprofen * Long term meloxicam: 0.03-0.1mg/kg PO 3x/week or up to every other day with close monitoring * Carprofen - single dose 2-4mg/kg IM,IV,PO * Ketoprofen: less cox 2 specific, 1.5-2mg/kg IM/IV/SC * Others * Ketamine - CRI during procedure at low doses (10-40mcg/kg/min) and titrated now to 3-5 at end and stopped before waking them up * Alpha 2 * Local Anesthesia * Local blocks: head (tooth trauma) * Small cats: 0.1-0.3ml of 2% lidocaine or 0.5% bupivacaine * Large cats: 1-3ml * (A) Infraorbital nerve- rostral part of upper dental arcade, palates nose and muzzle (infraorbital foramen) * Inferior alveolar nerve * (B) Mental foramen - incisors, chin * C Mandibular foramen - entire lower dental arcade * Ophthalmic * topical prilocaine - 2 to 3 hrs * Retrobulbar nerve block for enucleation * Ophthalmic block - lower risks * Ear * Auriculotemporal nerve - rostral caudal to vertical ear canal * Epidural injection * L7-S1 * Preservative free morphine 0.1-0.3mg/kg and bupivacaine 0.3mg/kg (not exceeding 6-9mL * 8-12 hrs of analgesia, anesthesia 1-4 hrs * If thoracic or front leg analgesia use morphine and saline * Brachial plexus block - foreleg distal to and including elbow joint * High brachial plexus block - proximal at spinal canal * Distal three point block - amputations, onychectomy, etc * Dorsomedial surface of carpus just proximal to joint (radial nerve) * Either side of carpal pad blocks palmar and dorsal branches of ulnar nerves and median nerve

Answer 7

**Fowler 9, Chap 76: Update on Field Anesthesia Protocols for Free-Ranging African Lions** Chemical immobilization is fundamental for conservation (capture/translocation, treatment) * **Cyclohexylamine dissociatives** (phencyclidine, ketamine, tiletamine)- TZ most common, combos w/ reversible/partially reversible Capture Technique * Capture at night (more active/willing to approach)- make all preparations before nightfall: * Two sites (capture site, processing site) 25-200m apart to reduce contact * Capture site: open, cleared of vegetation for unobstructed view, large enough for vehicle * Processing site: large enough to accommodate personnel, lions, equipment, well lit, surrounded by vehicle barrier * Capture with partially eviscerated carcass dragged behind vehicle to create trail and fasted to tree/stake * Play recorded lion/hyena/dying animal sounds * Dart from vehicle, determine depth with painful stimulus before approach, blindfold, hobble forelimbs for transport * Monitor CV/R function, temperature, depth frequently, observe complete recovery and protect from attack **Agent:** **Effect** **Adverse Effects** **Reversal** **Tiletamine + Zolezepam** **TZ** (Zoletil, Telazol) Cyclohexylamine + Benzodiazepine \**drug of choice\** Rapid induction Good muscle relaxation Gradual, predictable recovery (up to 4 hours- dose dependent)- *vulnerable* More potent than ketamine= less volume Wide safety margin Limited CV/R effects * Tonic convulsions* * Hyperthermia* **Tiletamine + Zolezepam + Medetomidine** **TZM** Cychlohexylamine + Benzodiazepine + Alpha-2 Agonist Reduces TZ dose up to 75% Smooth induction 3-10 minutes Good muscle relaxation Sufficient analgesia for minor procedures Spontaneous recovery after 1 hour -repeat IM TZM 1/3 induction dose PRN (could result in prolonged recovery) Limited stable CV/R depression Hg saturation \>90% ***Hyperthermia*** May retain palpebral reflex Partial: **Atipamezole** **-**reduces recovery time (8-30 min if given 45 min after induction) **Ketamine + (Me)Detomidine** Cyclohexylamine + Alpha-2 Agonist Induction 6-10 minutes Good for short-duration procedures (radio-collaring, disease surveillance, snare removal) Limited CV/R effects Hg saturation 85-90% Partial: **Atipamezole** **-**25-32min when given 1 hour after induction **Butorphanol + Medetomidine + Midazolam** **(BMM)** Opioid + Alpha-2 Agonist + Benzodiazepine Rapid induction 5-10 min 45 min stable anesthesia Sufficient for minor procedures- additional analgesia needed for surgery Spontaneous recovery after \>45 min - administer IM BMM 1/3 induction dose after 45 min and every 30 min - loud noise, painful stimulus- use hobbles/blindfold Mild bradycardia (\<50 bpm) Mild metabolic acidosis Normocapnia PaO2 ~ 87 mmHg **Complete: Naltrexone + Atipamezole + Flumazenil** - standing 4-8min - can be given at any time - no adverse effects seen when given IV - give additional atipamezole PRN (10x medetomidine dose) Helicopter Darting * Capable pilot, experienced vet * Marked sympathetic response- 2x+ TZM/BMM doses to be effective; TZ drug of choice (**hyperthermia**) * Approach with caution until depth confirmed

Answer 8

**Non-Infectious Diseases: Nutritional** * Cranial malformation * Young (\<2y) African lions * Incoordination, ataxia, head tilt, star-gazing, opisthotonus * Low serum and hepatic vitamin A * Unrewarding treatment but improvement with vit A in mild cases * Genetics? **Fowler 7 Ch 61 - Stargazing in Lions** * Caused by proliferation of cranial bones with resulting compression of brain tissue in young and adolescent captive African lions * No gender predilection * No reports in Asian lions * Rare historical reports in a 10-month old tiger and in leopards **Clinical Signs** * First observed in subadult lions (9 to 14 months) * Rare cases signs observed as early as 2 months of age * Early detection important in potential treatment during bone formation * Can affect one littermate while the others develop normally * Most often a combination of neurological signs * Neuroanatomic expression of a peripheral or central vestibular disorder * Slow progression of signs over weeks- best to take repeated video * Initial signs- * Ataxia, lack of coordination, and difficulty negotiating obstacles * Affected animal eats normally, but smaller amounts * Often smaller than unaffected littermates * Later signs- * Progressive ataxia, mild head tilt and cycling behavior (most often left-sided), stargazing, nystagmus, fine head tremor, staring glare, unresponsiveness to new objects. * Animal becomes lethargic and depressed * Ataxia worse when animal excited * Single cases- abnormal vocalization, hypersalivation, and tongue protusion * End stage- * Blindness, convulsitons, inability to stand, rolling over, recumbency, death **Diagnosis and Differential Diagnoses** * Complete PE, neuro exam * Bloodwork- * CBC/Chem, Vit A, FIV, coronavirus, feline leukemia * Screen for *Toxoplasma gondii, Ehrlichia canis*, Lyme, RMSF * Cerebrospinal fluid sampling- virology, micro, parasitology * PCR for canine distemper, protein and cell counts * Imaging (rads, U/S, MRI, CT) * Thickening of os tentorium cerebelli * Herniation of caudal folia through foramen magnum occipitalis * Compare max diameter of vitreous body of eye to thickest part of occipital bone and os tentorium cerbelli * Hyperintensity of spinal cord consistent with syringohydrom yelia and secondary enlargement of lateral ventricles from stasis of CSF * Liver biopsy for hepatic Vit A levels * Rule out- * Trauma, infection (coronavirus, FIV, canine distemper), otitis, space-occupying process (hematoma, abscess, neoplasia, congenital, pathologic bone) * Serum Vitamin A Concentrations in Lions * Liver is reservoir organ for vitamin A * Serum vitamin A level used for diagnosis, therapy, comparison between littermates * Congenital deficiency of lipoproteins needed for endogenous vitamin A transport therorized * Parenteral vitamin A supplementation is limited in the short-term * \*\*Serum vitamin A levels below 60 ug/L are critical for development of stargazing in growing lions * Values higher than 90 ug/L are adequate and should be maintained in captivity **Cause and Pathogenesis** * Exact cause is uncertain * First ascribed to vitamin B deficiency * Suggested that bony changes result from low vitamin A stores during growth * May not see changes when hypovitaminosis A occurs later in life * Vitamin A reported to stimulate osteoclast activity and resorb bone * Skeletal deformity from defective remodeling of membranous bone * Inadequate resorption of endosteal bone * Causes asynchrony between developing CNS and bones of skull and spinal column * Cranium- * Bones of caudal fossa severely affected * Cerebullum may herniate into foramen magnum * Other membranous bones may be affected- eg. Long bones * Endochondral bone does not appear to be directly influenced by vit A deficiency * Hydrocephalus caused by impaired absorption of CSF is another possibility * CSF absorption occurs in arachnoid villi located in tentorium cerebelli * Hypovitaminosis theory refuted in a study of 149 cases of affected lions and leopards in France * Primary genetic issue not discovered through pedigree analysis * Infectious viral cause proposed as cause **Treatment** * Glucocorticoids to reduce brain swelling and edema * Vitamin A supplementation * Start with parenteral since may have disorder of enteral vit A absorption * Some reports of improvement, but CNS signs often recur and worsen * Presumed that treatment started early when bones growing respond better * Suboccipital craniectomy and laminectomy- * Two cases reported to surgically decompress cranial fossa resulted in resolution of clinical signs * May be the only way to cure the condition * Individual in advanced state of disease should be euthanized immediately **Necropsy and Histopathology** * Severe thickening of osseum tentorium cerebelli and occipital bone * Subsequent herniation of caudal cerebellar folia through foramen magnum occipitalis * Increased thickness of all bones of the skull and mandibular bones * Dilation of lateral ventricles and syringomyelia of cervical spinal cord * Histo lesions of bones- * Thickened, poorly remodeled bone tissue caused by a shift from compact to cancellous bone * **Tentorium cerebelli**- * Thickening caused by growth of new periosteal woven bone containing retained cartilaginous cores * Histo of nervous system * Herniated cerebellar folia, compressed brainstem, and cervical spinal cord * Cerebellar lesions- * Thinning and rarefaction of the molecular layer, loss of Purkinje cells, granular cells associatd with proliferation of Bergmann’s glia, and disseminated punctate hemorrhages * Varying degrees of malacia and wallerian degeneration- * Dilated myelin sheaths, axonal swelling (spheroids), and digesting chambers associated with astrogliosis in white matter of compressed cerebellar folia, medulla oblongata, and tracts of cervical spinal cord * Edema and syringomyelia mostly in dorsal tracts of cervical spinal cord * Meningeal lymphoplasmacytic infiltration, fibroplasia, and hemorrhages in compressed regions **Prevention** * Careful evaluation and supervision of diet of African lions, esp. growing cubs for adequate vitamin A levels * Cases in this report received adequate vitamin A, so other factors must be considered * Pathologic alterations in vitamin A absorption or metabolism have been considered * Enzyme carotenase is absent in felids * Further studies of vitamin A digestion, absorption, and metabolism in lions needed * Daily liver feeding not recommended since could result in diarrhea or hypervitaminosis

Answer 9

**Non-Infectious Diseases: Toxic** * Mercury (monomethyl and inorganic) * Florida panthers, exposure via predation of piscivorous prey (raccoons) in the Everglades

Answer 10

**Non-Infectious Diseases: Congenital/Genetic** * Atrial septal defects and cryptorchidism – pumas and Florida panther – inbreeding * Multiple ocular colobomas (defect in one or more eye structures) – snow leopards * Unilateral or bilateral upper eyelid * Polycystic kidneys – Pallas’ cats * Adults, confined to kidneys, genetics?

Answer 11

**Non-Infectious Diseases: Age-related/Degenerative** * Spondylosis (spondylosis deformans, ankylosing spondylosis) * Asymptomatic or decreased activity, weakness, ataxia, paresis * Lumbar spine and lumbosacral junction most common

Answer 12

**Non-Infectious Diseases: Miscellaneous** * Veno-occlusive disease – cheetahs and snow leopards * Progressive collagen accumulation in perisinusoidal spaces 🡪 loss of hepatocytes 🡪 fibrosis (cirrhosis in chronic cases) * Primary lesion is subendothelial damage * Hepatic failure * Dietary – phytoestrogens or elevated vitamin A? * Amyloidosis – cheetahs and black footed cats * Black footed cat * Deposition in renal medulla, glomeruli, spleen, and GIT * Amyloid associated renal failure young (~4yr) * Not associated with chronic inflammation, genetics? * Cheetahs * Amyloid fibrils deposited in renal medulla -\> papillary necrosis * Deposits contain full serum amyloid A and fragmented AA protein * Concurrent inflammation (gastritis) \*\*Unlike black footed cats\*\* * Glomerulosclerosis – cheetahs * Glomerular basement membranes, thickened with sclerotic glomeruli, no glomerular inflammation * Parallels lesions in diabetic nephropathy of humans except cheetahs are not diabetic * Oxalate nephrosis – cougars, jaguar, leopards, cheetahs * Dead or acute renal failure with severe azotemia * Calcium crystals in urine, basement membrane can become mineralized * Birefringent crystals forming rosettes present in renal tubules * Ethylene glycol toxicosis suspected in some cases but never proven * No association between risk factors (concurrent renal disease or gastritis) and severity * Most severe cases in young cheetahs * Leukoencephalopathy – cheetahs * First diagnosed 1996, no new cases since 2005 * Older, ataxia, suspected blindness, disorientation, seizures * Suspect neurotoxin in food, drugs, or vaccines * Cavitation within the white matter and hydrocephalus * Myelopathy – cheetahs * 2.5 months to 12 yrs; rapid and fatal * Ataxia, paresis 🡪 dragging hindlimbs * Copper deficiency? * Endometrial hyperplasia * More common in melengesterol acetate contracepted animals * Develops earlier, increased risk with increased treatment duration (\>72months), age increased risk, parity decreased risk * Mild to marked expansion of endometrium with or without cysts * Severe cases with endometrial fibrosis or stromal mineralization

Answer 13

Zoo Path * Leukoencephalopathy – cheetahs * First diagnosed 1996, no new cases since 2005 * Older, ataxia, suspected blindness, disorientation, seizures * Suspect neurotoxin in food, drugs, or vaccines * Cavitation within the white matter and hydrocephalus * Myelopathy – cheetahs * 2.5 months to 12 yrs; rapid and fatal * Ataxia, paresis -\> dragging hindlimbs * Copper deficiency? **Fowler 6 Ch 32 - Neurologic Disorders in Cheetahs and Snow Leopards** * Captive diseases * Don’t forget CDV, spondylosis, IVDD in your differentials **NEUROLOGIC DISEASES IN CHEETAHS** **_Cheetah myelopathy_** * GENERAL * Spinal cord, degenerative lesions -\> ataxia and paresis * 60 cases; 25% of all deaths * Course: variable. Progressive ataxia to slower development including stabilization and acute relapsing episodes * CLINICAL SIGNS * Cubs mostly, adults too but not as common * Onset: peracute; related to stressful event * Herpesvirus (FHV-1) concurrent infection is typical * Pelvic limb paresis is first; proprioceptive problems, hypermetria, ataxia, recumbency * Disuse atrophy of hindlimbs * Variable course of disease * EPIDEMIOLOGY * 60 cases * Europe, United Arab Emirates * Captive born animals; same parents will have normal litters * No common denominator for housing, diet * Vaccination is common, but disease occurs before vaccination in many cases * Temporal association with Herpesvirus infection in cubs (NOT DEFINITIVE) * DIAGNOSTICS * Rads, myelography and MRI were WNL * CSF WNL * CBC/chem. WNL * Liver copper no difference b/w normal and ataxic animals * FIP, CDV, FPV, FCV, FeLV, FIV, Borna disease virus (BDV), EMC, tick borne encelphalitis, mucosal disease complex virus, Teschen-talfan disease virus, Listeria, Chlamydophila * FHV-1 and toxo positive in some cases and not others * “Classic” cheetah diseases on necropsies * Gastritis, glomerulonephritis/sclerosis, hepatic/renal amyloidosis, myelolipoma * NO correlation with myelopathy * PATHOLOGY * Lesions in spinal cord are rare * White matter affected exclusively: degeneration, ballooning of myelin * See text for greater detail * THERAPEUTIC TRIALS * No treatments recommended * Many tried * Supportive, some animals survive, recurrence is high * ETIOLOGY * NOT reported in North America; South Africa or Japan * NOT familial, but maybe genetic * Ddx: genetic, nutritional (esp copper), toxic, viral **_Cheetah leukoencephalopathy_** * North American cheetahs * NOT Europe or So Africa * Clinical signs: blindness, lack of response to environment, behavioral change, incoordination, convulsions * Started in 1996, peaked in 2001, declining now * N=70 animals * Adults \>10 y * Path: cerebral cortex, white matter * Unknown etiology, epidemiology suggests diet or medical management * Diagnosis: MRI, histopath * Irreversible **_Feline spongiform encephalopathy_** * Prion disease, spongiform encephalopathy related to BSE * In other exotic felids (ocelot, tiger, lion, cougar, puma) but higher prevalence in cheetahs * N=9 * Adults \>5y * UK, France * Clinical signs: chronic progressive ataxia starting in hindlimbs; progresses to other neurp signs * Diagnosis: histopath * Etiology: BSE from food **_Other neurologic disease observed_** * Vitamin A deficiency * ‘Coning’ of cerebellum * Ischemic necrosis of spinal cord **NEUROLOGIC DISEASES IN SNOW LEOPARDS** * 2 distinct spinal cord disorders * Finland, Switzerland, France * Etiologies unknown; genetic suspected * France (1 litter): * Cubs. Parents had successful litters previously. * Tremors/swaying gait, paresis. * Histo: myelin sheath dilation, axonal dilation * France/Switzerland/Finland (3 litters): * Cubs * swaying gait, hypermetria, hindlimb weakness. * No gross lesions. * Histo: loss of myelin.

Answer 14

**Fowler 7 Ch 60 - Aging in Large Felids** **Table 60-1-** good chart listing effects of anesthetic agents on aged felids **Table 60-2**- List of common complications in chronic renal failure and possible treatments **AGING** * Aging likely combination of evolution, genetics, metabolic damage, cellular senescence, and toxin accumulation * Phenotypic changes seen with aging- pelage, skin, or body condition * Internal physiologic changes- less obvious * Free-ranging animals- alterations frequently result in individual succumbing to disease, predation, or starvation * Captive animals- protected, but therefore see more degenerative age-related diseases * Jaguar study- increase in incidence and variability of disease processes w/aging **HEALTH CARE IN CAPTIVE FELIDS** * Vague signs of illness (e.g., reduced appetite, weight loss) may be noted in age-related diseases * Early detection/treatment will improve, possibly prolong, quality of life for certain conditions * Dental disease, osteoarthritis, chronic renal failure * AAHA guidelines for senior domestic cats can be applied to large felids * Routine PEs, sample collection, imaging should be performed as part of preventative medicine * Training for voluntary sampling helps avoids risk of anesthesia * Aged individuals more likely to have underlying chronic disease or degenerative organ dysfunction that may increase risk **COMMON DISEASES** * Renal system and urinary tract * Chronic renal disease significant cause of morbidity/mortality in old exotic felids * Common causes: chronic pyelonephritis, glomerulosclerosis, amyloidosis * Clinical signs: PU/PD, anorexia, poor pelage, salivation, oral ulceration, wt loss, emesis * Diagnosis: blood and urine analysis * Prognosis: poor long-term b/c changes usually irreversible * Treatment: slow down progression and alleviate clinical signs * Musculoskeletal * Osteoarthritis * Vertebral, elbow, stifle joints most common * Concrete substrate increase susceptibility * Degenerative spinal disease * Clinical signs: reduced activity, chronic intermittent hind limb paresis, ataxia * C/s show up 10-19y of age * Diagnosis: radiography (also CT/MRI, but not in textbook) * Intervertebral disc mineralization or herniation, spondylosis, spinal cord damage * Treatment to optimize quality of life * Pain control (NSAIDs), weight reduction, nutritional supplements, environmental manipulation * NSAIDs contraindications- renal or hepatic insufficiency, dehydration, hypotension, conditions affecting circulatory volume (such as congestive heart failure or ascites), moderate or severe pulmonary disease, and gastric ulceration * Side effects- GI ulcerations, nephropathies, and impaired coagulation * Reproductive * High prevalence of leiomyomas and leimyosarcomas * MGA or repeated sterile matings w/pseudopregnancies lead to uterine changes * Can lead to endometrial hyperplasia, uterine carcinomas, mammary neoplasia, pyometra, ovarian cysts * Mammary carcinoma rapidly metastasizes (lymph nodes, lung, liver) * Ovariectomy protects from mammary carcinoma but not benign tumors * Cardiovascular and respiratory disorders * Important if older animal is too be anesthetized, but nothing specific to large cats * Chronic bronchial disease, pulmonary neoplasia, myocardial and degenerative valvular disease and systemic diseases (e.g., hypertension w/hyperthyroidism or chronic renal disease) all reported * Neurologic disorders * Senescence can increase physiologic and behavioral susceptibility to environmental stressors * Behavior can change in older large felids, just like in older domestic cats * Beta-amyloid and abnormalities of tau phosphorylation seen in aged large felid brains * Leukoencephalopathy in cheetahs * Slowly progressive, degeneration and necrosis of cerebral cortical white matter * Most affected animals \>10y old * Irreversible, supportive treatment only * MRI may be helpful for antemortem diagnosis of neurologic conditions, but most often postmortem * Neoplasia * Common in older animals in general * Benign conditions can progress to malignancy (e.g., viral papillomatosis and SCC in snow leopards) * Nutritional disorders * Obesity- genetics, lack of physical acitivity, diet, and female contraception * Exacerbates musculoskeletal problems, predisposes to neoplasia, metabolic derangements (diabetes, hepatic lipidosis) * Protein, phosphate restrictions increase median survival time of cats with chronic renal disease * Dental disease * Significant cause of morbidity * Captive get calculus and abscesses (wild get attrition and fractures) * Other conditions * Thyroid adenocarcinoma, but no reports of hyperthyroidism * Inflammatory GI diseases * Chronic liver disease (e.g., neoplasia, cirrhosis, toxic degeneration) * Supportive treatment and prognosis is poor * Ocular disease frequent (due to e.g., chronic dz, neoplasia, degeneration, systemic dz) * Retinal detachment/hemorrhage from hypertension * Insular amyloidosis (maybe assoc. w/type II diabetes) * Oral eosinophilic granulomas in tigers 8-16 yrs old * Feline spongiform encephalopathy- develops over several years and pathology and clinical signs not present until animals are older

Answer 15

**Non-Infectious Diseases: Neoplastic** * Splenic myelolipomas – cheetahs * Mature adipose with hematopoietic cells, incidental and benign * Transitional cell carcinomas – fishing cats * Pheochromocytomas – clouded leopards * Squamous cell carcinomas – snow leopards * Associated with papillomavirus infection * Leiomyomas * Older females; predisposed to hydrometra and pyometra * Uterine and mammary carcinomas * Progestin-based contraceptives (MGA) **Common Neoplasms in Felids** * Biliary cyst * African and Asian lions * Single, multiple and mulilocular * Cysts in the kidney, mutations in *polycystic kidney disease 1* not identified * Majority do not have concurrent renal cysts * Biliary adenomas and adenocarcinoma less common; lions and leopards * Lymphoma * Older (\>14yr) African lions * T cell predominately * Not associated with FeLV or FIV in lions * Cheetah with FeLV and multicentric T cell lymphoma * Multiple myeloma * African lion, jaguar, snow leopard * Similar disease in domestic animals * Spreads to extramedullary sites * Leiomyomas and leiomyosarcoma * Common in female genital tract * More common in uterus than ovary or broad ligament * Leiomyomas can occur in cervix * Leiomyosarcoma less common than leiomyomas * Progestin contraception (long-term treatment) with MGA associated with mammary gland carcinogenesis * Non-significant association with leiomyosarcoma and MGA treatment * Transitional cell carcinoma * Fishing cats (10.8yrs old, no sex predilection) * Trigone of the urinary bladder * Treatment with COX-2 inhibitors may slow progression * Captive cats fed beef & little fish, increased saturated fatty acids, dietary? * Recommend decrease beef and increase fish in diets * Ovarian papillary cystadenocarcinoma * Jaguar * Significant mortality in females affecting breeding age and geriatric * Diagnosis is common at necropsy * Multiple cystic masses, bilateral, spread along uterine horns and abdominal carcinomatosis * Differential: peritoneal mesothelioma * No correlation with progestin (MGA) contraception * Pheochromocytoma * Clouded leopards; uncommon * Splenic myelolipoma * Cheetahs * Mammary gland carcinoma * Many species; jaguars overrepresented (genetics?) * Older animals, aggressive, metastasis * More commonly positive for progesterone receptors than estrogen * Increased incidence with long term (\>3 yr) MGA contraception * Concurrent exposure to estrogen and progestin as predisposing factor * Felids are seasonally polyestrus, induced ovulators; if not bred peaks of estrogen and progestin as a factor for carcinogenesis * May be associated with hypercalcemia or anemia * Endometrial hyperplasia * Spontaneous or MGA associated (most common in MGA treated animals)

Answer 16

**Treatment of Chronic Renal Failure in Nondomestic Felids (Fowler 6 Chapter 57)** * significant cause of morb/mort * caused by chronic pyelonephritis, glomerulosclerosis, amyloidosis * treatment aimed at slowing progression and improving quality of life _Diagnosis_ * clinical signs: PU/PD, decreased appetite, wt loss, poor pelage, ptyalism, oral ulceration, emesis, dehydration * work up: PE, CBC, chem, UA, kidney biopsy, fractional clearance, urine culture, +/- coag panel * clin path * mild nonregenerative anemia +/- neutrophilia * increased BUN, creat, P, chol * decreased Ca, K, alb, TP * normal USG or not * UPC 0.4-0.5, \>1 is significant proteinuria * fractional excretion of K, Ca, P, Na * these and UPC change before significant increases in BUN and creat _Treatment_ * directed against primary cause, if possible * discontinue nephrotoxic drugs * rule out other dz conditions * fluid therapy * more likely to drink if water is filled frequently and kept cool * ice cubes with meat juice or chicken broth in water * add water to diet (e.g., in whole prey) * training/conditioning for SQ fluids * nutritional support * decreased protein diet, but make sure adequate calories and protein * omega-3 polyunsaturated fatty acids à decrease BP, +/- renal protective effect * introduce commercial feline renal diets (nondomestics rarely will eat this as 100% of diet) * keep cat eating (goal: 70 kcal/kg/day) and monitor wt * try to stabilize wt, creat, alb * potassium supplementation * PU and tubular acidosis increase excretion, anorexia decreases intake * avoid hyperkalemia (rare if cat is PU) * phosphorus binding * decreased renal excretion * metastatic calcification when Ca-P exceeds 70 mg/dL * reducing dietary intake difficult * e.g., Epakitin, Ampojel * pulse antibiotics * 2-3wk quarterly for pyelonephritis _Proteinuria_ * monitor microalbuminuria/microproteinuria as well * ACE inhibitors à vasodilation in nephron à reduced hydrostatic pressure à less protein forced thru _Hypertension_ * see all the above * add beta blocker if ACE inhibitor not enough _Uremia_ * see all the above * metaclopramide for emesis * H2-blocker or PPI plus sucralfate for gastric erosion and small ulcers

Answer 17

* Herpesvirus * Mild signs – sneezing, nasal discharge, inappetence, ocular lesion * Vaccination with modified live vaccine may induce disease * Severe disease with corneal ulcers & keratitis in Pallas’ cats and cheetahs * Cheetahs develop unusual skin lesions due to *feline herpesvirus 1* * Ulcerations and plaque like proliferative lesions on face, forelimbs, or sites in contact with lacrimal and salivary glands * Ineffective immune responses may play a role in susceptibility * Biopsy needed to rule out neoplasia * Feline rhinotracheitis virus (Feline herpesvirus 1) * Highly contagious, shed in salvia, ocular, and nasal secretions, spread on fomites * High morbidity, low mortality, self-limiting, co-infections common * CS: ocular discharge, conjunctivitis, blepharospasm, sneezing, nasal discharge, keratitis, ulcerative dermatitis in cheetahs. * May become chronic carriers

Answer 18

* Parvovirus * Single stranded * Susceptible to both *feline panleukopenia virus* and *canine parvovirus 2* * Breeding or housing facilities particularly prone despite vaccination in late pregnancy using an inactivated vaccine * Hardy virus in cool, moist, shaded environments * Reduced litter size, severe lymphoid depletion with or without cerebellar or intestinal lesions (necrohemorrhagic enteritis) * Diagnosis: fecal electron microscopy, IHC, or VI * Disease via canine parvovirus may be milder than feline panleukopenia * Feline Panleukopenia virus (Parvovirus) * Highly contagious, shed in all secretions * CS: fever, depression, anorexia, dehydration, vomiting, diarrhea, causes panleukopenia * Virus resistant to inactivation, can survive \>1 year in environment

Answer 19

* Papillomavirus * Non-enveloped, circular, family *Papillomaviridae* * Most are species specific and specific tissue tropism * Development role of oral and cutaneous papillomas \> SCC and sarcoids * Basilar epithelial cells as primary site of infection * Oral papillomas – snow leopards, ventral tongue * Sharp line of demarcation between lesion and non-lesion * Rare inclusions; confirm disease via IHC, ISH, EM, or PCR * Squamous cell carcinoma * Common in slow leopards * Oral SCC may be due to papillomavirus * Sublingual tumors * Sarcoids * African lions – perioral masses; pumas – between lip and nasal planum

Answer 20

* *Canine distemper virus* * Bronchointerstitial pneumonia and non-suppurative encephalitis * Disease in wild felids due to unvaccinated domestic dogs * Exposure and infection is widespread in wild felids

Answer 21

* *Feline enteric coronavirus* * Feline infectious peritonitis (FIP) * High protein effusions, pyogranulomatous inflammation * Epidemic in captive cheetahs in 1983 (\>60% mortality) * Feline Coronavirus (coronavirus group 1) * Enteric coronavirus (infects intestines) & feline infectious peritonitis virus (FIP) * Highly contagious, shed in feces, fecal-oral route, * CS: (FIP) fever, vomiting, diarrhea, effusions with high protein content * Not considered directly transmissible from cat to cat, depends on virus mutation and low immunity.

Answer 22

* *Feline calicivirus* * Self-limiting infection; highly contagious * Vesicles and ulcer on tongue, oral and nasal mucosa * Disease occurred after vaccination with modified-live vaccines * Feline calicivirus (calicivirus) * Highly contagious, shed in salvia, ocular, and nasal secretions, spread on fomites * Sneezing, ocular and nasal discharge, oral ulcers of the gingiva and tongue * Animals that recover may shed virus for months to years

Answer 23

* Influenza type A virus * Avian influenza, *Orthomyxoviridae* family * Aerosolized respiratory secretions and feces * Clouded leopards, leopards, and tigers died of high-path AI in 2003-2004 after being fed infected chicken and quail * High fever, respiratory distress

Answer 24

* *Feline immunodeficiency virus* * Most extensively studied in African lions * Controversy if disease is associated with infection * One FIV-associated lymphoma case described * Different subtypes associated with different mortality rates * Feline immunodeficiency (Lentivirus) * Shed in saliva, transmission through bites, endemic in certain lion populations * CS: often asymptomatic, can include oral cavity disease, anemia, skin infections, weight loss, vomiting, diarrhea or neurologic disease. * Easily inactivated by common disinfectants * Feline leukemia virus (Retrovirus) * Shed in saliva, tears, urine, semen, vaginal fluids, and feces * Oronasal contact with saliva or urine most common mode of transmission, vertical transmission possible * Contact with or ingestion of feral cats (Cheetah, Iberian lynx, cougar) * CS: immunosuppression, anemia, chronic inflammatory conditions, enlarged lymph nodes, persistent fever, lymphoid or myeloid tumors, repro problems. * Some cats remain persistently viremic * Unstable outside host * *Feline leukemia virus* * Endemic in free-ranging wildcats * Increased contact with domestic cats? * Mortality in Iberian lynx, Florida panthers, and a mountain lion; lymphoma in a cheetah * Viremia linked to anemia, lymphopenia, lymphadenopathy, sepsis, death

Answer 25

* *Bluetongue* virus serotype 8 * Eurasian lynx fed stillborn or aborted fetuses with BT infections * Anemia, SQ hematomas, petechiation, pulmonary congestion, and edema * Biting midges transmission?

Answer 26

* *Mycobacterium bovis* * African lions, leopards, cheetah, Iberian lynx, and bobcat * Cheetah, leopards, lynx – spillover hosts * Lions – maintenance hosts * PPE during necropsy (seroconversion of staff and keepers) * Infection from prey * African animals - buffalo; lynx - cattle, boar, or fallow deer * Concurrent FIV infection plays no role in pathogenesis in lions or lynx * Increased susceptibility or recrudescence related to in-breeding in lions * Signs: Emaciation, decubital ulcers, cutaneous alopecia, corneal opacity, poorly healed bite wounds, elbow/hock hygromas * Acid-fast bacteria common in airways, lungs, and joints; rare in pulmonary macrophages * Non-tuberculous mycobacteria: oropharynx of Argentinian healthy pumas

Answer 27

* Anthrax (*Bacillus anthracis)* * Gram positive, endospore forming, OIE reportable, zoonotic * Ingestion of infected prey * Titers in lions correlated with high rainfall * Cheetahs highly susceptible to fatal disease; exposure in lions may be protective (high mortality after periods of low incidence) * Infection through skin defects, ingestion, or inhalation of spores * Death due to edema, shock, acute renal failure, and CNS anoxia * Mild and transient disease in felids – head and neck swelling * Differential: cytotoxic snake bite * Disposal: incinerated, rendered, or buried * Stain slides with Giemsa stain – basophilic blunt ended rods with well developed capsules * Positive silver stain * Culture bacteria from unstained blood, tissue, or exudates

Answer 28

* Heicobacter sp. * Stomach incidental finding; rare gastritis in most felids (except cheetah) * Cheetahs \>95% associated with lymphoplasmacytic gastritis * Stomach thickened, reddened, cobblestone, loss of rugae * Signs: weight loss to chronic vomiting * Develop secondary amyloidosis

Answer 29

* *Salmonella* sp. * Can be shed from asymptomatic cats * *S. typhimurium* seen in \>90% of cats (from contaminated feed) * Plague (*Yersinia pestis)* * Eating infected rodents * Intralesional Gram negative coccobacilli

Answer 30

**Infectious Diseases: Fungi** * Cryptococcosis (*Cryptococcus gattii)* * Described in cheetah; inhalation of basidiospores * Ulcerative skin nodules * Mucinous capsule; stains with Alcian blue and Mayer’s mucicarmine * Dermatophytosis (*Trichophyton mentagrophytes* or *Microsporum* spp.) * Alopecia, pyoderma, hyperkeratosis * Topical treatments may be effective * Recommended in cheetah – griseofulvin associated with lethargy, diarrhea, and bone marrow suppression

Answer 31

**Infectious Diseases: Parasites** * *Ollulanus tricuspis* * Gastric trichostrongylid * Pumas seropositive for FeLV – infection predisposition? * Signs: vomiting, inappetence, bloody or mucoid diarrhea, ill-thrift * Life cycle completed in stomach (small nematode size, 1mm long) * Differentiate from *Helicobacter*-associated gastritis in cheetahs * *Toxoplasma gondii* * Rarely associated with disease * Pallas’ cats uniquely susceptible to infection – high neonatal mortality * Neonates: necrotizing encephalitis, pneumonia, and/or hepatitis * Adults: encephalomyelitis, recrudescence of latent infections? * *_Cytauxzoon felis_* -protozoan that infects blood cells * Transmitted by lone star tick (*Amblyomma americanum*) * Natural reservoir = bobcat (carrier after developing mild/subclinical infection) * Disease may be fatal in untreated domestic cats, fatal infection in a white tiger * *_Babesia felis_* -originally ID in wild cats from Sudan, reported sporadically in other countries * South Africa only country where feline babesiosis is a significant clinical entity * Infection assumed to be tickborne (vector has not been identified) * Ascarids (*Toxocara* and *Toxascaris*) common in captive animals; often difficult to eliminate * Ova highly resistant in the environment. * Repeated testing and treatment of animals while in quarantine recommended * *Dirofilaria immitis* (heartworm) - diagnosed in a black-footed cat * -Prophylactic treatment is recommended in endemic areas.

Answer 32

**REPRODUCTION (Fowler 8):** * All felids have induced ovulations, but some have spontaneous ovulations. * Occurs freq. in clouded leopards, fishing cats, and margays * rare in cheetahs, tigrinas, ocelots * Photoperiod sensitivity: * Pallas’ cats- very sensitive * Tigers, clouded leopards, snow leopards moderately affected * Ocelots, tigrinas, margays, lions, leopards, fishing cats not influenced by photoperiod. * Clouded leopards and Pallas’ cats exhibit seasonality in gonadal activity * Margays, cheetahs, and oncillas cycle year round. * Suppressed ovarian activity and estrus occurs in cats housed in a group (e.g., **_cheetah_**s). * All cats have a zonal placentation **AZA WILDLIFE CONTRACEPTION CENTER (2012) RECOMMENDATIONS FOR FELID CONTRACEPTION:** * GnRH agonists- considered the safest reversible contraceptives * Dosages and duration of efficacy are not well established for all species * Caution in lions due to prolonged response with questionable reversibility * Side effects similar to those assoc. with gonadectomy (potential for weight gain). * ex: Suprelorin (deslorelin) implants (female or male) * ex: Lupron® Depot injection (female or male) * Ovariohysterectomy or ovariectomy (females) or castration (males)-permanent sterilization * Progestin contraceptives * associated with progressive uterine growth, infertility, infections, mammary/uterine cancer * If progestin is used, tx should be short term, started before any signs of proestrus. * Progestins should not be used in pregnant animals.

Answer 33

**Fowler 6 Ch 35 – Use of Analgesics in Exotic Felids** Edward Ramsay NSAIDS * Inhibit cyclooxygenase (COX) enzymes in arachidonic acid metabolism * COX-1 – homeostatic processes (inhibition produces side effects) * COX-2 – response to noxious stimuli and the creation of proinflammatory prostaglandins which is associated with pain * Aspirin * Only NSAID with safe chronic dosage in cats * Anti-inflammatory, antiplatelet, and analgesic effects – not great for severe pain * Cats lack the ability to bind glucuronides so they do not metabolize and excrete salicylates well * Must be given at less frequent intervals * Vomiting and gastric hemorrhage are common side effects * Ketoprofen * Propionic acid derivative – analgesic and anti-inflammatory properties * COX-1 preferential * Good analgesia but reduces platelet activity and isn’t recommended after surgery * Short term (5-day) use has been investigated, but longer-term use not studied * Carprofen * Propionic acid derivative * COX-2 preferential agent * Reports of a leopard dying with intestinal ulcers after 2 weeks of use * Meloxicam * Enolic acid compound – analgesic, anti-inflammatory, antipyretic activities * COX-2 preferential * Few adverse effects reported * Piroxicam * COX-1 preferential agent * Anti-inflammatory, analgesic, and some antineoplastic properties * Narrower therapeutic window in dogs, less info on cats Opioids * Historic reluctance to use this class based on dysphoria, mania, and excitement. Other side effects include respiratory depression, loss of thermoregulatory ability, sedation, vomiting, mydriasis. * Large cats appear more sensitive than smaller cats * Morphine, oxymorphone, hydomorphone, meperidine * Mu-receptor agonists * Morphine associated with dysphoria or excitement * Fentanyl * Transdermal delivery offer \>72 hours of relief – 6-8 hour lag time after application * Patches only use in select cases based on animal temperament * Buprenoprhine * Partial mu agonist * Long duration of action, transmucosal application * Butorphanol * Mu-receptor antagonist, kappa agonist * Good analgesia to visera, but poor muscular analgesia * Short acting \<2 hours * Tramadol * Not an opioid, but has affinity for mu receptors * Inhibits reuptake of serotonin and norepinephrine within the spinal cord * Some dose-dependent respiratory depression has been observed Treatment of Acute Pain * Injectable regimens often better as cats may not eat following surgery or injury * Opioids have faster onset of analgesia than NSAIDs * Preoperative administration provides better results than postoperative * Still should consider the potential renal effects of NSAIDs and renal blood flow * One study showed that ketoprofen provided better analgesia following OVH than buprenorphine or meperidine * Another buprenorphine was better than oxymorphone or ketoprofen * Another showed NSAIDs were better than butorphanol * Still consider local and regional analgesia Treatment of Chronic Pain * Asprirnig 10 mg/kg PO q72h for arthritis administration * Meloxicam has been used in a variety of felids * Tramadol appears effective, has some sedation and ataxia at higher doses * Etodolac – indole acetic acid derivative (COX-2 selective) – has been used for arthritis in tigers, but there is a report of gastric ulcers * Adjunctive therapies including weight reduction, nutraceuticals, rearranging the exhibit should also be considered Treat to Effect – not every cat will respond the same way * Buprenorphine, meloxicam appear good for acute pain * Meloxicam and tramadol appear good for chronic pain

Answer 34

Journal of Zoo and Wildlife Medicine 51(3): 652-656, 2020 **HISTOLOGIC EVIDENCE OF SPONTANEOUS OVULATION IN TIGERS (*PANTHERA TIGRIS*)** Megan L. Cabot, DVM, Edward C. Ramsay, DVM, Dipl. ACZM, Deborah Chaffins, and Mee-Ja M. Sula, DVM, Dipl ACVP **Abstract**: Spontaneous ovulation has been identified in several wild felid species and domestic cats previously thought to undergo only induced ovulation. Two studies have assessed ovulation patterns in tigers (*Panthera* *tigris*) and have not found evidence of spontaneous ovulation in this species. However, **uterine pathology typically associated with prolonged progesterone exposure has been identified in unbred tigers, suggesting spontaneous ovulation occurs**. Ovaries from 47 tigers were reviewed with histologic examination. The presence or lack of active corpora lutea was documented and compared with social housing conditions for each animal. Social housing categories were as follows: female housed alone, female housed with other females, and female housed with at least one male. Active corpora lutea were identified in 66% (10/15) of females housed alone, 85% (6/7) of females housed with other females, and 58% of females housed with a male. A chi-squared test **found the presence of active corpora lutea was independent of social housing condition.** These results offer strong evidence of spontaneous ovulation in tigers. This finding suggests that **a luteal control protocol with an early follicular inhibition agent may improve assisted reproduction efforts and supports spaying nonreproductive or** **postreproductive female tigers to reduce the rate of uterine infections**. Key points: * Multiple felid species (lions, clouded leopards, domestic cats, Pallas’ calls) have been found to exhibit spontaneous ovulation despite the classic view that felidae species were exclusively induced ovulators * Cheetah, tigrinas, and ocelots found to be only induced ovulations * Pyometra, which occurs secondary to prolonged progesterone influence) has been described in at least one tiger that was housed alone * This study paired histologic evaluation of tiger ovaries with information about their social housing situation for 1 year prior to tissue collection * Presence of active corpus lutea was found to be independent of social housing condition * Higher incidence of active corpora lutea in females housed with other females compared to those housed with males or housed alone * Female-female contact may promote reproductive cycling and spontaneous ovulation in female species * Follicular inhibition protocol before gonadotropin administration may improve ovarian response to stimulation protocols for artificial insemination, which currently has a low success rate in tigers (\<5%) * Non-breeding tigers at risk for uterine pathology such as pyometra à recommend prophylactic ovariectomy or OVH Take home: Evidence to support spontaneous ovulation in tigers as active corpora lutea were found to be independent of social housing condition. This finding may impact AI protocols and management of non-breeding individuals.

Answer 35

Chinnadurai, S. K., Kinsel, M. J., Adkesson, M. J., & Terio, K. (2017). Canine distemper in a vaccinated snow leopard (Panthera uncia). *Journal of Zoo and Wildlife Medicine*, *48*(4), 1200-1203. Abstract: A 6-yr-old male snow leopard (*Panthera uncia*) **presented with acute seizures, hyperthermia, and tachypnea. Because of a diagnosis of anuric renal failure, the animal was euthanized.** On histopathologic examination, **numerous intralesional intracytoplasmic and intranuclear inclusions were found in the lungs, lymph nodes, and stomach. Positive immunohistochemical staining for canine distemper virus (CDV)** was found in the lungs and, to a lesser extent, in the lymph nodes and brain. Molecular testing yielded a CDV H gene sequence that was closely related to CDV isolates concurrently found in wild raccoons from adjacent forested areas. **The leopard had been vaccinated once against CDV with the use of a recombinant canarypox-vectored live vaccine during a routine wellness examination 12 wk prior to death. Serial serum neutralization titers performed on banked serum collected between vaccination and death showed poor serologic response to the vaccine. This case demonstrates a probable failure of protection against naturally occurring CDV.** * Canine distemper virus – morbillivirus, family Paramyxoviridae. * Globally important dz of canidae, felidae, procyonidae, mustelidae, viverridae. * Dogs – GI, resp, CNS, skin dz. * Felids may have no resp or GI signs. * Current reocmmendations for nondomestic felids – vx with a recombinant canarypox-vectored CDV vaccine if risk of exposure is high. * In this report, animal had seizures 12 wks after routine exam/vaccines. * Looked at banked serum samples, this animal did not mount a serologic response to the vx in the 6 wks post vaccination. * Serologic titers do not represent the entire picture of vaccine-induced immunity as they exclude the role of CMI. * Infection suspected to have come from raccoons, other large cats in the collection that were not vaccinated had serologic evidence of exposure.

Answer 36

Lin, Y. W., & Wang, L. C. (2018). Animal training and acupuncture in a Bengal tiger (Panthera tigris tigris) with hind limb paraparesis. *Journal of Zoo and Wildlife Medicine*, *49*(2), 493-496. Abstract: An 18-yr-old, captive-born male Bengal tiger (*Panthera tigris tigris*) presented a 1-yr history of chronic and **progressive paraparesis** in both hind limbs. Lateral and hind limb radiographs were revealed normal except for **severe spondylosis deformans**, forming a bony bridge between the last lumbar and the first sacral vertebra and disc mineralization between the second and third lumbar vertebra**. Medical therapies were instituted, including corticosteroids, hydroacupuncture and electroacupuncture. Animal training allowed veterinarians to perform acupuncture** safely without having to anesthetize the animal. Animal training made intensive treatment possible. **Neither corticosteroids nor hydroacupuncture alone provided much clinical improvement. The tiger reacted positively after electroacupuncture was performed. The tiger began showing clinical improvement after three electroacupuncture treatment**s and could eventually walk on all four limbs at the end of the treatment. * Electroacupuncture – Activates pain inhibition system, improves nerve blood flow in patients suffering from spinal disorders. * Acupuncture points – spondylosis and mineralized disc locations: * Lumbar Baihui, left and right BL 23, BL 25, BL 32, and BL49. * LB – Facilitates repair of neuronal tissues assoc with pelvic limb paresis or paralysis. * BL 23, 25, 32, 49 – treat lumbar pain, lower extremity paralysis, lumbar sacral pain. * This case report – 2 weeks of training, blunt capped needles, blunt needles with tip cut off, then sharp needles. Showed some improvement on prednisolone 0.4 mg/kg PO SID, hydroacupuncture with injections of vit B1 and B12 performed into 6-7 points, no improvement. * After 6 months, tried electrostimulator with acupuncture point locator. * Performed right after the hydroacupuncture on tx days. * Performed every other day for 8 tx and then twice weekly for 12 tx. * Improved after first 3 treatments, was able to walk on all four limbs most * of the time after 2 mos. Takeaway: Acupuncture techniques combined with animal training was safe, effective alternative to sx in large captive felid with paraparesis.

Answer 37

**SARS-COV-2 INFECTION AND LONGITUDINAL FECAL SCREENING IN MALAYAN TIGERS (PANTHERA TIGRIS JACKSONI), AMUR TIGERS (PANTHERA TIGRIS ALTAICA ), AND AFRICAN LIONS (PANTHERA LEO KRUGERI) AT THE BRONX ZOO, NEW YORK, USA** Bartlett, Susan L., Diel, Diego G., Wang, Leyi, Zec, Stephanie, Laverack, Melissa, et al. _Abstract:_ Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) emerged as the cause of a global pandemic in 2019–2020. In March 2020, New York City became the epicenter in the United States for the pandemic. On 27 March 2020, a Malayan tiger (Panthera tigris jacksoni) at the Bronx Zoo in New York City developed a cough and wheezing with subsequent inappetence. Over the next week, an additional Malayan tiger and two Amur tigers (Panthera tigris altaica) in the same building and three lions (Panthera leo krugeri) in a separate building also became ill. The index case was anesthetized for diagnostic workup. Physical examination and bloodwork results were unremarkable. Thoracic radiography and ultrasonography revealed a bronchial pattern with peribronchial cuffing and mild lung consolidation with alveolar-interstitial syndrome, respectively. SARS- CoV-2 RNA was identified by real-time, reverse transcriptase PCR (rRT-PCR) on oropharyngeal and nasal swabs and tracheal wash fluid. Cytologic examination of tracheal wash fluid revealed necrosis, and viral RNA was detected in necrotic cells by in situ hybridization, confirming virus-associated tissue damage. SARS-CoV-2 was isolated from the tracheal wash fluid of the index case, as well as the feces from one Amur tiger and one lion. Fecal viral RNA shedding was confirmed in all seven clinical cases and an asymptomatic Amur tiger. Respiratory signs abated within 1–5 days for most animals, although they persisted intermittently for 16 days in the index case. Fecal RNA shedding persisted for as long as 35 days beyond cessation of respiratory signs. This case series describes the clinical presentation, diagnostic evaluation, and management of tigers and lions infected with SARS-CoV-2 and describes the duration of viral RNA fecal shedding in these cases. This report documents the first known natural transmission of SARS-CoV-2 from humans to nondomestic felids. *Journal of Zoo and Wildlife Medicine, 51(4) : 733-744* Key Points: * Signs of SARS-COV-2: dry to wet cough, sneezing +/- anorexia +/- mild vomiting * Most resolved within 5 days * Likely acquired from the keepers * Diagnosed on tiger with PCR of oronasal swab and tracheal cytology * Other tigers and lions diagnosed and monitored with fecal PCR * Fecal PCR shedding lasted for up to 35 days after resolution of clinical signs. * Conclusions: Tigers and lions caught SARS-CoV-2 from keepers and were diagnosed and monitored with fecal PCR. _Introduction:_ * SARS-COV-2- Shares 96% homology with a coronavirus isolated from bats (BatCoV RaTG13) * NYC epicenter of the pandemic around March 2020 _Case series:_ * Increase in staff members working at both of the exhibits that house the lions and the tigers (normally would have separate staffing) * See below for case information for each animal affected * All animals treated with clavamox BID x 14 days * Index case had persistent coughing at day 6- immobilized for treatment + diagnostics * Normal vitals, oxygen saturation * Tracheal wash- epithelial necrosis, mixed inflammation * rRT-PCR presumptive positive for all swabs- tracheal, oropharyngeal, nasal; genome sequenced- aligned with SARS-COV-2 obtained from humans; viral neutralization assay titer of 1:64 * Report of diagnostics + sample methodology: McAloose D, et al. From people to Panthera: natural SARS-CoV-2 in tigers and lions at the Bronx Zoo. 2020. mBio. 2020;11(5):e02220-2 * CXR- bronchial pattern with multifocal caudal peribronchial cuffing; thoracic ultrasound- small areas of consolidated lung * CBC/chem- normal WBC count, lymphocytes reactive * Treated with cefovecin SQ, LRS SQ, penicillin SQ, cerenia * Complete resolution of coughing by day 16 * Daily fecal collection on the 5 tigers + 3 lions started on day 8- all positive for passing SARS-CoV-2 RNA; asymptomatic tigers in the other exhibit were all negative * Index case shed viral RNA for 14 days * Asymptomatic tigers shed variably- 5 days up to 24 days * Prolonged shedding in two lions for more than 30 days * Genome sequencing of the viral RNA showed that the tigers and lions were infected with different SARS-CoV-2 genotypes - infected in unrelated transmission events * No keepers had clinical signs, but tested positive for COVID * Safety measures put in place- masks, distancing, gloves, eye protection, separate coveralls/diet prep/cleaning tools, gentle hosing only after cleaning with Rescue _Important points from discussion_: * Infection not definitely confirmed in all felids- did not want to immobilize to collect respiratory tract samples * Presumed infection based on clinical respiratory signs, fecal viral RNA shedding and virus isolation from feces of one lion and one tiger * Two domestic cats in NY state documented to have naturally acquired SARS-CoV-2 about the same time as the tiger/lion infections * Detection of RNA in feces via PCR indicates presence of noninfectious viral RNA fragments, intact viable virus or both (vs. virus isolation confirms the presence of viable virus) * In humans- RNA shedding in feces persists beyond 3 weeks and after cessation of clinical signs * Prolonged fecal viral RNA shedding, recovery of viable virus in the feces and the spontaneous recurrence of fecal RNA shedding after prolonged cessation of respiratory signs → evidence for active replication of virus in the GI tract * GI clinical signs in two of the cases - after respiratory signs stopped but still shedding RNA in feces * Currently no signs of long term complications for the felids

Answer 38

MAST CELL TUMORS IN CHEETAH (ACINONYX JUBATUS): A CASE SERIES Journal of Zoo and Wildlife Medicine 51(4): 1025–1034, 2020 Abstract: Mast cell tumors in nondomestic felids are rarely reported and their biological characteristics are not well described**. A retrospective review of the pathology records of 52 zoo-housed cheetahs (Acinonyx jubatus) identified five cases of mast cell tumor, involving four closely related individuals**. The age at initial presentation varied from 14 mo to 6 yr. Four cases presented as solitary or multiple cutaneous masses that were mostly slow growing, up to 20 mm diameter, and predominantly nonulcerated. The diagnosis was made by fine needle aspiration cytology of a lesion in one case and by excisional biopsy in the others. Histopathologically, the lesions resembled low- to intermediate-grade canine mast cell tumors, with variations in the degree of anisocytosis and anisokaryosis. **Surgical excision was incomplete for 80% of the cutaneous lesions, but local recurrence was not observed in any case. One animal with cutaneous lesions subsequently developed fatal visceral mastocytosis** involving the spleen, liver, and adrenal gland. There was no evidence of lymph node invasion or paraneoplastic gastrointestinal signs in any of the cases. Intro * Mast cell tumors (MCT) are uncommonly reported in nondomestic felids * Retrospective review of path records of 52 zoo-housed cheetahs from 1981-2016 Case reports * 5 cases of mast cell tumor identified, involving 4 closely related individuals * The age at initial presentation varied from 14 mo to 6 years * Slow growing cutaneous masses, either solitary or multiple * Predominantly nonulcerated * All initially diagnosed by FNA * All low to intermediate grade based on histopath * Surgical excision was incomplete for 80% of the cutaneous lesions, but local recurrence was not observed in any case * One animal with cutaneous lesions subsequently developed fatal visceral mastocytosis involving the spleen, liver, and adrenal gland. * There was no evidence of lymph node invasion or paraneoplastic gastrointestinal signs in any of the cases. Discussion * Mast cells have previously been described as a feature of chronic eosinophilic dermatitis resulting from herpesvirus infection in cheetahs. However, the features in the cases reported here do not support that diagnosis. * No epidermal hyperplasia or herpesviral inclusions were found * Grading scheme is variable, but subjectively , the histological features of the tumors more closely resembled those seen in dog MCT, rather than those in domestic cat MCT * 4/5 were incompletely excised, supporting the fact that the tumors often extend beyond the visible mass * However, recurrence was still not observed up to 6 years post op and/or on necropsy * The occurrence of visceral lesions in case 1 many years after incomplete excision of a cutaneous MCT may have represented metastatic spread from the original cutaneous nodules or de novo visceral mastocytosis (the latter is suspected) * Possible genetic component due to the relatedness of these animals

Answer 39

Journal of Zoo and Wildlife Medicine 51(3): 720-724, 2020 **HISTOPLASMOSIS IN NONDOMESTIC FELIDS: A REVIEW OF SIX CASES** Anthony J. Cerreta, DVM, Denae N. LoBato, DVM, PhD, Dipl ACVP, Edward C. Ramsay, DVM, Dipl ACZM, and Andrew C. Cushing, BVSc, MRCVS, Cert AVP (ZM), Dipl ACZM **Abstract**: **One adult leopard (*Panthera pardus*) and five adult tigers** (*Panthera tigris*) presented with a range of **nonspecific clinical signs**, including lethargy (6/6), mobility deficits (4/6), and hyporexia (3/6). Hematology and biochemistry revealed a **hyperproteinemia characterized by hyperglobulinemia** (4/6), hepatocellular enzyme activity increases (3/6), azotemia (3/6), leukocytosis (2/6), hyperbilirubinemia (2/6), or a combination of conditions. Further diagnostics and management varied with the presenting signs and clinicopathological findings, including supportive care, diagnostic imaging, and blastomyces urine antigen analyses. **Two animals died, and four were euthanatized**. Postmortem findings included **granulomatous pneumonia (6/6),** fibrinous pleural effusion (3/ 6), pericardial effusion (2/6), and diffuse icterus (1/6). Histopathology revealed round to oval structures with a thin clear wall and **purple inclusions within cells of the mononuclear phagocyte system, consistent with *Histoplasma* *capsulatum*,** in each animal. **Disseminated histoplasmosis was found in five cases**, with organisms present in the **lung (5/5), liver (3/5), lymph nodes** (3/5), spleen (2/5), bone marrow (2/5), thyroid (1/5), tongue (1/ 5), kidney (1/5), or a combination of organs. One tiger was found to have pulmonary histoplasmosis without evidence of disseminated infection. On the basis of clinical and pathological findings, histoplasmosis was diagnosed. This case series illustrates the **difficulties in antemortem diagnosis of histoplasmosis** on the basis of complete blood count, serum biochemistry profile, and antigen testing and underscores that histoplasmosis should be considered a **differential diagnosis in any felid presenting with nonspecific clinical signs in endemic areas.** Key Points: * Soil-borne, free-living, dimorphic fungus; highest prevalence in midwestern and southern states (Ohio, Missouri, and Mississippi river) * Second most common systemic fungal disease in domestic cats – chronic disease and nonspecific * Previously reported in a tiger and two snow leopards – died shortly after onset of signs * Five of the cases occurred in 2019; all housed with dirt and turf substrates – suspect environmental * Leopard: 10yr FI, leukocytosis, azotemia, hyperproteinemia, and elevated liver enzymes * Mild peritoneal effusion, fluid cytology consistent with *H. capsulatum*; died during recovery * Severe diffuse pyogranulomatous hepatitis, splenitis, and lymphadenitis * Tigers: MC, 10-20yrs; neutrophilic leukocytosis, hyperproteinemia, azotemia, and increased LEs * One died post-anesthesia, remaining 4 euthanized * Multifocal chronic pneumonia in all cases * *H. capsulatum* within the lungs, highlighted with PAS stain * All demonstrated lethargy with onset from 1 week to 1 month; majority also had hyporexia Take home: Lethargy and hyporexia, leukocytosis with hyperproteinemia and elevated liver enzymes, narrow based budding yeast, difficult to diagnose antemortem, significant morbidity and mortality

Answer 40

Anderson, K. M., Garner, M. M., Clyde, V. L., Volle, K. A., Ialeggio, D. M., Reid, S. W., ... & Wolf, K. N. (2018). **Gastric dilatation and enterotoxemia in ten captive felids.** *Journal of the American Veterinary Medical Association*, *253*(7), 918-925. **Abstract**: CASE DESCRIPTION: 10 large felids at 8 facilities were determined or suspected to have developed gastric dilatation with or without enterotoxemia over a 20-year period. Four felids were found dead with no premonitory signs. CLINICAL FINDINGS: 4 felids (2 male snow leopards [*Uncia uncia*], 1 male Amur tiger [*Panthera tigris altaica*], and 1 male Sumatran tiger [*Panthera tigris sumatrae*]) were found dead or died before they could be evaluated. Six felids had hematemesis (1 male and 1 female African lion [*Panthera leo*] and 1 male jaguar [*Panthera onca*]) or abdominal distention and signs of lethargy with or without vomiting (1 male African lion, 1 male Malayan tiger [*Panthera tigris jacksoni*], and 1 female Sumatran tiger). Gastric dilatation was radiographically and surgically confirmed in the male Malayan and female Sumatran tigers and the jaguar. TREATMENT AND OUTCOME: In 3 felids with an antemortem diagnosis, the gastric dilatation resolved with decompressive laparotomy but then recurred in 1 felid, which subsequently died. Three others died at various points during hospitalization. Although *Clostridium perfringens* type A was recovered from 3 of the 5 felids for which microbial culture was performed, and 2 felids had a recent increase in the amount fed, no single factor was definitively identified that might have incited or contributed to the gastric dilatation. CLINICAL RELEVANCE: Gastric dilatation was a life-threatening condition in the large felids of this report, causing sudden death or clinical signs of hematemesis, abdominal distention, or vomiting. Even with rapid diagnosis and surgical decompression, the prognosis was poor. Research is needed into the factors that con-tribute to this emergent condition in large felids so that preventive measures might be taken. * 10 felids (lion, leopard, tiger, and jaguar) diagnosed with gastric dilatation +/- volvulus * 4 felids found dead without premonitory signs * 2 snow leopards had increased 20% diet * Amur tiger possible clostridium A * 6 felids had hematemesis, other signs abdominal distension, lethargy, vomiting * Decompression unsuccessful and exploratory laparotomies performed in 5 felids * One lion passed acutely before evaluation * Most developed life-threatening complications, 3 felids survived, 2 died in surgery, 1 was euthanized * Recurrent gastric dilation occurred with one surviving tiger which died from complications (assumed adipose tissue + serosal fibrosis 🡪 delayed gastric emptying) * All 3 lions showed histological or gross evidence of gastric dilatation that spontaneously resolved, reason lions are prone unknown * Possible predisposing factors * C perfringens may have contributed to two tigers at same facility * Residual bromine in exhibit water features for jaguar (once eliminated vomiting stopped) * Gastric FB: trichobezoar in lion, bones in tiger * Food volume increase in snow leopards found deceased **Take** **home**: Gastric dilatation is life threatening condition that requires surgical decompression, prognosis poor

Answer 41

McDermid, K. R., Snyman, A., Verreynne, F. J., Carroll, J. P., Penzhorn, B. L., & Yabsley, M. J. (2017). Surveillance for viral and parasitic pathogens in a vulnerable African Lion (Panthera Leo) population in the Northern Tuli Game Reserve, Botswana. *Journal of wildlife diseases*, *53*(1), 54-61. **Abstract:** African lion (Panthera leo) numbers are decreasing rapidly and populations are becoming smaller and more fragmented. Infectious diseases are one of numerous issues threatening free-ranging lion populations, and low-density populations are particularly at risk. We collected data on the prevalence and diversity of viral and parasitic pathogens in a small lion population in eastern Botswana. During 2012 and 2014, blood samples were collected from 59% (n¼13) of the adult-subadult lions in the Northern Tuli Game Reserve in eastern Botswana. **One lion had antibodies to feline panleukopenia virus, two had antibodies to canine distemper virus, and two had feline calicivirus antibodies. Ten of the 13 had antibodies to feline immunodeficiency virus and 11 had feline herpesvirus antibodies**. All lions were negative for antibodies to feline coronavirus. Blood samples from all lions were negative for Trypanosoma, Anaplasma, Theileria, and Ehrlichia spp. by molecular testing; however, all lions were positive for Babesia spp. by reverse line blot hybridization assay. **Sequencing of amplicons from four lions revealed four groups of Babesia spp. including several genetic variants of Babesia felis, Babesia lengau, and Babesia canis and a group of novel Babesia sequences which were only 96% similar to other Babesia spp**. Six lions were infested with four species of ticks (Rhipicentor nuttalli, Rhipicephalus simus, Rhipicephalus sulcatus, and Rhipicephalus appendiculatus). These data provide the first health assessment of this population and can be used to identify management and conservation strategies to decrease the impact of pathogens on this population. This is particularly important as there is an initiative to incorporate this population into a larger metapopulation of lions from adjacent South Africa and Zimbabwe. * Interaction between Babesia & CDV important in 1994 distemper outbreak * All lions positive for Babesia * Host to B. felis, B. leo, B. lengau, and B. gibsoni * Antibodies to FCoV, FCV, and FPV rare or absent * Cub survival during FCV outbreaks are lower * High prevalence of FIV & FHV consistent with other studies * FHV not often associated with clinical disease however **Take-Home:** Lions – Babesia is common

Answer 42

Quintard, B., Greunz, E. M., Lefaux, B., Lemberger, K., & Leclerc, A. (2017). Squamous cell carcinoma in two snow leopards (uncia uncia) with unusual auricular presentation. *Journal of Zoo and Wildlife Medicine*, *48*(2), 578-580. **Abstract**: Squamous cell carcinoma (SCC) is well documented in snow leopards (Uncia uncia) and most common locations are oral, facial, or pedal. These two cases illustrate an unusual auricular presentation, which is more often reported in white domestic cats. The animals were aged and presented clinical signs of otitis such as head shaking and ear scratching. Clinical examinations showed auricular canal masses with chronic purulent otitis. In both cases, clinical deterioration led to euthanasia and histology of the ear canal was consistent with SCC and showed numerous vascular emboli. **These cases illustrate an unreported aggressive localization for SCC in snow leopards, which should be included in the differential diagnosis of otitis in this species. Auricular SCC may be underdiagnosed as the ear canal is infrequently sampled for histopathology**. This auricular localization should be considered when metastases are found upon necropsy without internal primary tumor. * Case Reports * Two cases, each presenting with otitis externa, masses or polyps observed * Initially cultured and treated with antibiotics – both declined later and were euthanized and metastasis was seen on necropsy * Papules or nodules in feline ears * Granulomas (infectious or tick associated), neoplasia –SCC most common - others include mast cell, basal cell, fibrosarcoma * Squamous cell carcinomas * Third most common tumor after mammary carcinoma and lymphoma * Main tumor in snow leopards – oral, facial, and pedal distribution common * Secondary to prolonged UV exposure in light colored animals * Pinnal SCC are known to be aggressive in many species * Associated with papillomavirus in snow leopards * Treatment * Topical imiquiomod – side effects & daily topical application makes it difficult * Surgery, radioation, electrochemotherapy **Take Home:** Sample weird lesions on the ears of snow leopards

Answer 43

Lara, V. M., Benassi, J. C., Bisetto, S. P., De Oliveira, T. M., Ferreira, H. L., Júnior, J. P. A., & Carregaro, A. B. (2017). **Molecular detection of infectious pathogens of the upper respiratory tract in captive nondomestic felids.** *Journal of Zoo and Wildlife Medicine*, *48*(2), 529-531. **Abstract:** Upper respiratory tract disease is a complex infectious disease process with multiple pathogens involved. Identification of infectious agents in wild animals is of great importance for wildlife conservation. The aim of this study was to evaluate the molecular detection of feline herpesvirus type 1, feline calicivirus (FCV), Bordetella bronchiseptica, Chlamydophila felis, and Mycoplasma felis using ocular and nasal swabs in three species of captive nondomestic felids. **Mycoplasma felis was detected in two ocular samples of Puma concolor and in one nasal sample of one Panthera onca**. FCV was detected in association with M. felis in one P. concolor. The other pathogens tested were not detected. To the authors’ knowledge, this is the first report of M. felis in nondomestic felids from Brazil. * Feline respiratory diseases * Feline Herpesvirus * Reported in wild lions, leopards, and puma * Feline calicivirus * Found in both wild and domestic species * Shed virus for longer than 30 days * Chlamydophila felis * Bordetella bronchiseptica * Mycoplasma felis * Considered commensal but can become pathogenic **Take Home:** Mycoplasma felis is likely commensal in multiple felid species

Answer 44

Bauer, K. L., Sander, S. J., Steeil, J. C., Walsh, T. F., & Neiffer, D. L. (2017). **Calcinosis circumscripta in a cohort of related juvenile african lions (panthera leo).** *Journal of Zoo and Wildlife Medicine*, *48*(3), 813-817. **Abstract:** Three juvenile, genetically related African lions (Panthera leo) were evaluated for discrete domeshaped subcutaneous masses present over the proximal lateral metatarsal-tarsal area. The lesions measured 3–8 cm in diameter, were ﬂuctuant to ﬁrm, nonulcerated, and attached to underlying structures. On radiographic evaluation, the lesions were characterized by well-circumscribed punctate mineralizations in the soft tissue surrounded by soft tissue swelling without evidence of adjacent bony involvement. On cut surface, the lesions were made of numerous loculi containing 2–5-mm round-to-ovoid, white-to-gray, ﬁrm structures interspersed with ﬁbrous tissue and pockets of serosanguinous ﬂuid. Hematology, serum biochemistry, serum thyroid screening (including total thyroxine, total triiodothyronine, free thyroxine, and free triiodothyronine), and serum vitamin D panels (including parathyroid hormone, ionized calcium, and 25-hydroxyvitamin D) were unremarkable. Histopathologic evaluation of the lesions was consistent with calcinosis circumscripta with ﬁbroplasia, chronic inﬂammation, and seroma formation. An additional two genetically related lions were considered suspect for calcinosis circumscripta based on presentation, exam ﬁndings, and similarity to the conﬁrmed cases. All masses self-regressed and were not associated with additional clinical signs other than initial lameness in two cases. * Calcinosis circumscripta background * syndrome of ectopic deposition of calcium salts in soft tissue structures * Occur most commonly on hind feet or tongue in animals * Different types classiﬁed  dystrophic, metastatic, iatrogenic, and idiopathic * Dystrophic calcinosis circumscripta * If normal Ca and P levels - suspected response to localized tissue damage * Metastatic calcinosis circumscripta - renal failure or vitamin D toxicosis * Often associated with hypercalcemia and/or hyperphosphatemia * Iatrogenic calcinosis circumscripta - has been associated with SC medroxyprogesterone and polydioxanone use in dogs * Idiopathic calcinosis circumscripta - no tissue damage or metabolic defect found * Calcinosis circumscripta discussion * May regress on own or require surgery depending on location, size, etc * All cases here regressed on own * Treated with NSAID’s * All lions were related in the case series * All lions were \<2yr old * Cause undetermined * Can be genetic in humans * locations of lesions were sites susceptible to repetitive trauma by play ﬁghting * Normal Ca and P levels combined with suspected inciting tissue damage suggests diagnosis of dystrophic calcinosis circumscripta

Answer 45

Cannizzo, S. A., Stinner, M., & Kennedy-Stoskopf, S. (2017). **Prevalence of cystinuria in servals (Leptailurus serval) in the United States.** *Journal of Zoo and Wildlife Medicine*, *48*(4), 1102-1107. Abstract**: Cystinuria is a condition caused by defects in amino acid transport within the kidneys and small intestines.** It has been reported in humans, dogs, domestic cats, ferrets, nondomestic canids, and nondomestic felids, including servals (*Leptailurus serval*). Genetic mutations have been identified in dogs, humans, and domestic cats. **Cystinuria usually follows an autosomal recessive inheritance, although it can be autosomal dominant and sex linked.** The primary objective of this study was to screen urine samples dried on filter paper from captive servals in the United States for cystinuria by using the cyanide-nitroprusside screening test. A second objective was to determine whether cystinuria is inheritable in servals. Servals were initially recruited for the study by survey. Owners and institutions interested in participating were sent a second survey and filter paper for collecting urine samples. Samples were collected from 25 servals**. One additional serval with confirmed cystine urolithiasis was added for a total sample size of 26 servals. Twenty-seven percent (7/26) were positive, 54% (14/26) were weakly positive, and 19% (5/26) were negative.** Sex, reproductive status, and urine collection method had no significant association with test results. **This condition is likely underreported in servals and should be ruled out in any serval with nonspecific signs of illness; neurologic signs such as lethargy, ataxia, or seizures; ptyalism; or signs of lower urinary tract disease such as dysuria, hematuria, stranguria, pollakiuria, or urethral obstructions.** **Key Points** * **Cystinuria – defect in transport of dibasic amino acids i.e. cysteine, ornithine, lysine, arginine in the epithelium of the proximal renal tubules and small intestines.** * **Healthy animals, these AA should be completely reabsorbed by proximal tubules of the kidneys.** * **Defects in AA transport system excrete high concentrations of these substances in urine.** * **Cysteine can precipitate into crystals and uroliths resulting in crystalluria, stranguria, dysuria, hematuria, urinary tract obstruction, renal failure.** * Primary objective of this study was to determine the prevalence of cystinuria in servals in the US and investigate lineages for evidence of inheritance. * Methods * Cyanide-nitroprusside test performed on urine on filter paper. * 25 samples collected. * 27% positive, 54% weak positive, 19 % negative. * No significant differences among test results and sex, repro status, or urine collection method. * The amount of protein, water, and salt in the diet can affect excretion and solubility of cysteine in urine. * Only one serval that tested positive had a history of uroliths, no other histories suggested cytinuria. * Cystinuria should be ruled out in any serval with dysuria, hematuria, stranguria, pollakiuria, or urethral obstructions and should also be a differential for neuro signs i.e. lethargy, ataxia, seizures, ptyalism. * **Tx to consider include increasing water consumption and using potassium citrate to alkalinize urine.** * Four servals with the same parents had results consistent with an autosomal recessive pattern of inheritance. * One positive, two weak positive, one negative. * Data insufficient to conclude definitive inheritance.

Answer 46

Whitten, C., Vogelnest, L., D'Arcy, R., Thomson, P., & Phalen, D. (2019). **A retrospective study of reported disorders of the oral cavity in large felids in australian zoos.** *Journal of Zoo and Wildlife Medicine*, *50*(1), 16-22. Abstract: Disorders of the oral cavity are conditions reported by veterinarians that impact the health and welfare of large felids in human care. There have been no studies documenting the prevalence of these conditions and species affected in Australian zoos. **A review of the medical records of lions (Panthera leo), tigers (Panthera tigris), cheetahs (Acinonyx jubatus), jaguars (Puma onca), snow leopards (Panthera uncia), Persian leopards (Panthera pardus saxicolor), and cougars (Puma concolor) from 10 Australian zoos and an online survey of zoo professionals from Australian and New Zealand zoos was performed to determine the recorded prevalence of disorders of the oral cavity in these species.** **Preliminary assessments were also made to determine if there was an association between the occurrence of tooth fractures and diet, feeding practices, species, sex, and age of the animal. The study also examined associations of these conditions with behavior, such as fighting, and husbandry practices, such as the provision of enrichment items.** The review found that tooth fractures were common in tigers and lions greater than 8 yr of age. Animal caregivers attributed this to animals chewing on large, hard pieces of bone in some instances, but this could not be verified. Instances of bones being lodged between canine teeth were observed and appeared to be related to the feeding of bones of inappropriate size. Based on these findings, it is recommended that guidelines for bone size fed be developed and that animals over the age of 8 yr receive regular dental examinations under general anesthesia. * **Disorders of the oral cavity identified: tooth fractures, focal palatine erosion (FPE), periodontal disease, gingivitis of unknown etiology (1 case), bones lodged on canine teeth** * **Tooth fractures most commonly in aged (\> 8 yo) tiers and lions compared to other species and ages.** * **Canines \> premolars \> incisors \> molars** * Most commonly associated with fighting, not associated with chewing on large bones * 50% required intervention: root canal \> extraction \> crown, all by dental specialist * Bones lodged in mouth - all cases between max and mand canines * 52% resolved on own; 48% required general anesthesia and removal * Appears that bones cut too short are more problematic than long bones. * Abnormal tooth wear, failure of tooth eruption, malpositioning of teeth not seen in this study * Calculus, gingivitis were rare, no odontoclastic resorption * Lower rate of Focal Palatine Erosion in Cheetahs in AU zoos **Takeaways** * Dental fractures most common oral cavity disorder in lions and tigers, most frequently older than 8 yo, most commonly canines * Suspect improved diet and dental prophylactic procedures contribute to reduced records of calculus, gingivitis * Bones cut short may predispose to bone lodging on canines.

Answer 47

Cushing, A. C., Ramsay, E. C., Newman, S. J., Hespel, A. M., & Sula, M. J. M. (2019). **Hypergammaglobulinemia and myeloma in five tigers (panthera tigris): clinicopathological findings.** Journal of Zoo and Wildlife Medicine, 50(1), 219-224. Five adult tigers (Panthera tigris) presented with a range of clinical signs, including paresis (2/5), lameness (2/5), ataxia (3/5), anorexia (5/5), and lethargy (5/5). Each tiger demonstrated elevated plasma globulin levels (7.8–14.8 g/dl; [reference interval 2–5.1 g/dl]) on routine biochemistry, confirmed as a monoclonal gammopathy using protein electrophoresis. Serum gammaglobulin concentration ranged from 5 to 7.5 g/dl, or 45.1–63.4% of total protein concentration. Azotemia was present in three tigers. Diagnostics and management varied with the presenting signs but included magnetic resonance imaging, radiography, chemotherapy, supportive care, and euthanasia. In each case, necropsy revealed a neoplastic plasma cell proliferation in the bone marrow and one or more extramedullary sites. Lytic lesions in the thoraco-lumbar spine were found in three animals, and one lesion was associated with spinal cord compression. Splenomegaly was present in 4/5 cases. Histopathology confirmed a plasma cell neoplasm in each case, and immunohistochemistry staining with multiple myeloma oncogene 1 (MUM1) was positive in each case. CD20 staining was performed in two cases and was positive in one. CD3 staining was performed in the same two cases, and was negative in each. Based on the clinical, gross, microscopic, and immunohistochemical findings, myeloma was diagnosed in all five tigers. * Key Points: * Monoclonal gammopathies – proliferation of single clone of B lymphocytes. * Assoc with plasma cell neoplasias in cats – lymphoma, myeloma, chronic lymphocytic leukemia, myeloma related dz (MRD). * Differentiation of plasma cell from other round cells with IHC multiple myeloma oncogene 1 (MUM1). * Hypercalcemia, azotemia, cytopenias, Bence-Jones proteinuria all assoc with poor prognosis in dogs. * Tx – Chemo can be attempted, not curative, MST 2 yrs. * Other reports in non-domestic felids – lion, jaguar, snow leopard * Case reports: 5 unrelated tigers diagnosed with myeloma at necropsy. * Antemortem dx myeloma – clinical findings, osteolytic lesions, monoclonal gammopathy, Bence-Jones proteinuria, bone marrow plasmacytosis. * Initially, marked serum hypergammaglobulinemia. * Ddx chronic inflammatory proceses. * Histo confirmed MUM1 positive proliferation of malignant plasma cells. * Osteolytic lesions in appendicular skeleton are rare in domestic cats, may be more common in noncomestic felids. * Factors assoc with poor prognosis – azotemia, Bence-Jones proteinuria. * Hypercalcemia, cytopenias, leukemia not seen in this series. Takeaway: Any adult nondomestic felid with vague CS and/or peripheral neuro deficits, this is a differential. Histo at necropsy should include use of MUM1 IHC staining for dx.

Answer 48

Kilburn, J. J., Backues, K. A., Kain, S., & Pucket, J. (2019). **Lip-to-lid transposition and husbandry management for severe bilateral eyelid coloboma in three snow leopards (panthera uncia).** *Journal of Zoo and Wildlife Medicine*, *50*(3), 688-695. _Abstract_: Three sibling snow leopard (Panthera uncia) cubs were evaluated shortly after birth and found to have bilateral eyelid colobomas involving .50% of the upper central and lateral lid margins. The cubs also had iris to cornea persistent pupillary membranes and developed varying degrees of keratitis caused by trichiasis and exposure. No fundic abnormalities were noted. Given the severity of the defects an extensive surgical correction was required. Prior to surgical correction, the cubs were separated from the dam daily and a human-animal socialization plan was enacted to prepare the cubs for the intensive postoperative care that would be required. Bilateral lip commissure to eyelid transposition surgeries were performed on one cub at a time at 2 wk intervals starting at 3 mo of age. Postoperative care was labor and time intensive and each cub was maintained at the zoo’s veterinary hospital for 11–21 days with daily supervised visits with the cub’s siblings. All cubs were successfully reintroduced together and with the dam at the end of each hospitalization. The surgical procedure yielded fully functional eyelids and an excellent cosmetic appearance in all three cubs. The planned consistent human interaction with the cubs before and during the postoperative period made the procedure and its intensive aftercare possible and was crucial to the successful outcome of the surgeries. Journal of Zoo and Wildlife Medicine 50(3): 688–695, 2019 Key Points * Eyelid agenesis = eyelid coloboma = congenital abnormality where a section of the eyelid does not form normally during development * Usually involve the upper lateral eyelid margin * Can also be associated with persistent pupillary membranes, retinal dysplasia, retinal/optic nerve colobomas * Size of the eyelid coloboma correlates with degree of clinical signs- trichiasis, exposure keratitis, KCS, corneal vascularization, ulcers * Treatment depends on the size of the defect, severity of clinical signs, animal temperament * Surgical correction for larger colobomas- simple primary closure, two-stage procedures, rotational grafting procedures such as the lip commissure to eyelid transposition (lip to lid transposition) * Lip to lid transposition- tries to provide a fully functional eyelid margin by rotating and sliding the oral commissure of the mouth up to become the new lateral canthus of the eye * Provides as many normal tissue layers as possible- skin, mucosa, mucocutaneous junctions and may include the parotid duct * End result- new eyelid margin, protect the cornea, sometimes ability to blink * Previous cases documented in snow leopards- no etiology proven, surgery has been performed with variable results * **Case Reports**: three snow leopards born in one litter, all diagnosed with varying degrees of severe eyelid colobomas * Plan enacted to separate the cubs from the dam intermittently to provide socialization with humans to allow for less stress associated with medical treatments pre-and post-surgery (started at 1 month old) * All cubs had 50-60% of the lid affected * Surgeries performed between 3-4 months of age (4.9-8kg) * Graft taken from lip commissure- partial thickness incision, 8-10mm of oral mucosa left attached to the lip graft, dissected posteriorly, rotated to align with the recipient site, sutured to the bulbar conjunctiva * Post-op care: * Nails trimmed and nail caps placed (to protect the suture), e-collars * Hospitalized for between 11-21 days, human companionship for at least 10 hrs/day * Also allowed socialization time with other cubs and dam * Complications- corneal opacity from suture rubbing, suture dehiscience, minor cosmetic defects **Take home points:** * Success of these surgeries due to pre-operative socialization and prep and extensive post-op, early diagnosis * Eyelid colobomas reported in snow leopards, can be successfully corrected with a lid-to-lip transposition

Answer 49

Kinney, M. E. (2020). **Single port laparoscopic falciform ligament collection in a cheetah (acinonyx jubatus).** *Journal of Zoo and Wildlife Medicine*, *50*(4), 997-999. Abstract: **A single incision laparoscopic system (SILS) was used to remove the falciform ligament of an adult male cheetah (Acinonyx jubatus) during routine diagnostic liver biopsy. Adipose tissue isolated from the falciform ligament was used to establish a mesenchymal stem cell culture. T**he use of a SILS port for liver biopsy and falciform ligament collection allowed for a large amount of fat to be collected from a small surgical incision and rapid postoperative recovery. This case expands the use of the single incision laparoscopy surgical technique beyond reproductive sterilization procedures in large cats. * Falciform ligament - remnant of embryologic ventral mesogastrium (conveys umbilical vein from umbilicus to liver in fetus) * Evaluated as a source of mesenchymal stem cells in humans * 4 yo male cheetah undergoing routine diagnostic liver biopsy, convenience falciform ligament harvesting for a mesenchymal stem cell culture * Anesthesia: ketamine 1.1 mg/kg, medetomidine 0.021 mg/kg, butorphanol 0.2 mg/kg, midazolam 0.1 mg/kg IM, maint with isoflurane, intubated, ventilator, perioperative meloxicam, famotidine, ampicillin sodium IV, maropitant SQ * Single Incision Laparoscopy System (SILS) on ventral midline * Allowed introduction of apparatus under direct visualization and pneumoabdomen (8 mmHg CO2) to be established before cannulas advanced into abdomen * Allowed removal of falciform ligament without extending incision (tissue \> 10 mm) * Would not need to drop ovaries in abdomen during ovariectomy prior to incision enlargement for ovary removal from the body * Limitations: technically challenging due to limited triangulation, minimal space external to body for maneuvering surgeon’s hands - can use articulating hand instruments, angled endoscopes, and hand instruments of variable lengths * Falciform ligament removed with sealer/divider to cauterize vasculature prior to cutting * Prominent vessel on midline at caudal aspect with multiple smaller branching vessels - origin not described in cheetahs **Takeaways** * SILS port insertion advantageous when tissue \> 10 mm is being removed from abdomen * Single lap systems should be considered for zoo patients if equipment and skill set are available

Answer 50

Journal of Zoo and Wildlife Medicine 51(2): 426–432, 2020 **CONSERVATION CHALLENGES: THE LIMITATIONS OF ANTEMORTEM TUBERCULOSIS TESTING IN CAPTIVE ASIATIC LIONS (*****PANTHERA LEO PERSICA*****)** Fieke Marije Molenaar, DVM, MSc, D Zoo Med, MRCVS, Paul D. Burr, BSc (Vet Sci) Hons, BVM&S, PhD, MRCVS, Benjamin M.C. Swift, BSc, M Res, PhD, Catherine E.D. Rees, BA, PhD, and Nicholas Masters, MA, VetMB, MSc, Dip ECZM, MRCVS – Reviewed by MM **Abstract**: Genetic diversity of captive wild animals can be enhanced by moving those individuals with valuable genes between collections and through introduction of a new pair from a range country. This requires movement of animals, which is inherent with disease risks, such as the introduction of pathogenic Mycobacterium sp. (MTBC) into a zoological collection. Decisions need to be made based on the outcome of perimovement disease screening using an array of tests, the majority of which are unvalidated in the species. **A pair of endangered Asiatic lions (*****Panthera leo persica*****) imported from India to the United Kingdom were screened for MTBC using the comparative intradermal tuberculosis (TB) test, the feline interferon-γ blood test, and the experimental bacteriophage assay.** Reactions on all three tests prompted screening of the three resident Asiatic lions using the same tests, all of which were negative for MTBC. Based on these test results, the decision had to be made to exclude the genetically valuable pair from the current collection. MTBC could not be identiﬁed using further tests, including culture and PCR on a bronchoalveolar lavage, on feces, or on postmortem tissues. **This case series highlights the usefulness of a control group when interpreting unvalidated test results for detection of MTBC, the value of training big cats for conscious blood sampling, and the practical implications of placing the comparative intradermal TB test in the eyelids, when dealing with a species that requires a general anesthetic for most hands-on interventions**. * TB in big cats * Spillover of bovine TB into cape buffalo from 1960-1980 * Transmission to carnivores through inhalation and ingestion – mongoose, baboons, cheetah, leopards, and African lions * TB diagnostics * Single intradermal TB test in the cervical area, antibody detection, culture, PCR, bacteriophage assay * Two Asiatic lions brought to UK (from Indian zoo) for their genetics, underwent extensive quaranatine & testing. ID test avian and bovine PPD in eyelids – resulted in significant swelling * Other tests submitted – interferon gamma release assay, cultures, PCR – live bacteria in MTBC in blood of both lions. Lions had gametes harvested for AI prior to euthanasia * A single intradermal TB test may result in some false positives but can be a useful tool in detecting disease initially * Serological testing useful in lions, but the disease process relies on cell-mediated response Take home: Diagnosis of TB in carnivores remains challenging antemortem, but using controls when evaluating suspect cases with the intradermal testing, interferon gamma

Answer 51

Smith, C. K., Seddighi, R., Zhu, X., Tepe, A. J., Ramsay, E. C., & Cushing, A. C. (2018). **Use of plethysmographic variability index and perfusion index to evaluate changes in arterial blood pressure in anesthetized tigers (Panthera tigris).** *American journal of veterinary research*, *79*(8), 845-851. OBJECTIVE To investigate use of the plethysmographic variability index (PVI) and perfusion index (PI) for evaluating changes in arterial blood pressure in anesthetized tigers (Panthera tigris). ANIMALS 8 adult tigers. PROCEDURES **Each tiger was anesthetized once with a combination of ketamine, midazolam, medetomidine, and isoflurane.** Anesthetic monitoring included assessment of PI, PVI, direct blood pressure measurements, anesthetic gas concentrations, esophageal temperature, and results of capnography and ECG. **Mean arterial blood pressure (MAP) was maintained for at least 20 minutes at each of the following blood pressure conditions: hypotensive (MAP = 50 ± 5 mm Hg), normotensive (MAP = 70 ± 5 mm Hg), and hypertensive (MAP = 90 ± 5 mm Hg).** Arterial blood gas analysis was performed at the beginning of anesthesia and at each blood pressure condition. RESULTS Mean ± SD PI values were 1.82 ± 2.38%, 1.17 ± 0.77%, and 1.71 ± 1.51% and mean PVI values were 16.00 ± 5.07%, 10.44 ± 3.55%, and 8.17 ± 3.49% for hypotensive, normotensive, and hypertensive conditions, respectively. The PI values did not differ significantly among blood pressure conditions. The PVI value for the hypotensive condition differed significantly from values for the normotensive and hypertensive conditions. The PVI values were significantly correlated with MAP (r = −0.657). The OR of hypotension to nonhypotension for PVI values ≥ 18% was 43.6. CONCLUSIONS AND CLINICAL RELEVANCE PVI was a clinically applicable variable determined by use of noninvasive methods in anesthetized tigers. Values of PVI ≥ 18% may indicate hypotension. **Background** * Dynamic indices: hemodynamic variables that have respiratory-induced changes * Changes in pulse pressure, systolic pressure, or stroke volume with changes in intrathoracic pressure during respiration are used to evaluate circulatory volume and guide fluid therapy in humans * Spontaneous inspiration: intrathoracic pressure decreases - increase in venous return to heart - increased stroke volume - increased cardiac output - increased PI and PVI * Positive pressure ventilation: increased intrathoracic pressure impedes venous return to the heart, decreasing stroke volume * Photoplethysmogram - waveform on pulse ox display * Indirect measure of blood volume or pulsatile strength at target tissue * Indices (PI and PVI) may be useful surrogates for blood pressure measurement * PI - indicator of peripheral blood flow; influenced by stroke volume and blood flow * % derived from relationship between pulsatile and nonpulsatile blood flow * PI = (AC/DC)x100; AC: pulsatile infrared signal; DC: nonpulsatile infrared signal * PVI - calculated from changes in PI by alterations in blood flow during the respiratory cycle * PVI = ([PImax - PImin]/PImax)x100 **Key Points** * Isoflurane used to induce hypotension; dobutamine used to achieve normotension and hypertension * PI did not significantly differ between blood pressure categories * No significant correlation between PI and MAP or PI and PVI * PVI for hypotension was significantly greater than for normotension and hypertension * Did not differ significantly between normotension and hypertension * Significant negative correlation between PVI and MAP * PVI _\>_ 18% significantly more frequently hypotensive * Same PVI cutoff for hypotension in dogs and children * No significant difference in heart rate or PaO2 between blood pressure categories **Conclusions** * PVI and MAP were significantly correlated in tigers * PVI for hypotension was significantly higher than normotension and hypertension in tigers * PVI _\>_ 18% significantly associated with odds of hypotension (MAP \< 60 mmHg) in tigers

Answer 52

**Performance of two portable blood glucose meters for measuring blood glucose concentration in tigers (Panthera tigris) and lions (Panthera leo)** JAVMA 2019 254(3) 399-408 **Abstract**: OBJECTIVE To investigate the performance of a portable blood glucose meter (PBGM) designed for use in humans (hPBGM) and a PBGM designed for use in dogs and cats (vPBGM) when measuring blood glucose (BG) concentration in tigers (Panthera tigris) and lions (Panthera leo). DESIGN Method comparison and diagnostic accuracy study. SAMPLES 53 blood samples from tigers (n = 27) and lions (26). PROCEDURES **BG concentration was measured with 2 identical hPBGMs, 2 identical vPBGMs, and a reference laboratory analyzer**. Bland-Altman bias plots and PassingBablok regression analysis were used to assess agreement. Sensitivity, specificity, and positive and negative predictive values with corresponding 95% confidence intervals were calculated for use in assessing diagnostic accuracy of the investigated PBGMs. RESULTS Bias (95% limits of agreement) was –4.3 mg/dL (–46.3 to 37.6 mg/dL) for the hPBGM, –9.3 mg/dL (–64.6 to 46.0 mg/dL) for the vPBGM on canine setting, and 2.3 mg/dL (–47.9 to 52.6 mg/dL) for the vPBGM on feline setting. The **hPBGM had better overall repeatability** (coefficient of variation, 3.73%) than the vPBGM on canine (9.29%) or feline (9.44%) setting. Total error for the hPBGM, vPBGM on canine setting, and vPBGM on feline setting was 11.8%, 27.7%, and 20.9%, respectively. **None of the PBGMs complied with the maximum allowable total error suggested by current guidelines when measuring BG in tigers and lions with hypo-, normo-, or hyperglycemia**. CONCLUSIONS AND CLINICAL RELEVANCE **Results indicated that the PBGMs evaluated were inadequate for measuring BG concentration in tigers and lions.** **Summary**: * vPBGM on canine setting had worst precision, agreement, and clinical reliability for detecting hypoglycemia in tigers and lions, but had acceptable sensitivity and specificity for detecting hyperglycemia in both species * hPBGM had ~6% better repeatability than vPBGM on canine (9.29%) or feline (9.44%) settings * clinical use of hPBGM in tigers and lions would not be recommended because 95% of results from hPBGM could differ from true BG concentration by ~ ± 40mg/dL and because sensitivity for detecting hypo- and hyperglycemia was 57.1% and 62.5% * overall PBGMs did not have good agreement with reference laboratory analyzer

Answer 53

Lamglait, B., & Vandenbunder-Beltrame, M. (2017). **Evaluation of symmetric dimethylarginine as an early biomarker of chronic kidney disease in captive cheetahs (Acinonyx jubatus). ** Journal of Zoo and Wildlife Medicine, 48(3), 874-877. Abstract: Symmetric dimethylarginine (SDMA) has been shown to be a valuable biomarker for early detection of chronic kidney disease (CKD) in canine and feline patients. Recognition of early (subclinical) kidney disease would be of value in cheetahs (Acinonyx jubatus) as prevalence of CKD is relatively high in this species in captivity. Fifty-eight banked serum and plasma samples from seven adult cheetahs that died of CKD were analyzed for creatinine, urea, and SDMA.** A marked increase in SDMA was noted on five of the tested cheetahs earlier than the rise of serum creatinine and urea (estimated 8–35 mo; mean 21.4 mo; median 22 mo). SDMA appears as an early biomarker to evaluate renal function for the diagnosis of CKD in cheetahs regardless of the cause of this disease.** Key Points: **SDMA – Symmetric dimethylarginine** * Derived from intranuclear methylation of L-arginine by methyltransferases and released into circulation after proteolysis. * Excreted primarily by renal clearance. * Accurate biomarker for estimating glomerular filtration rate (GFR) in humans.** Cheetahs prone to renal disease. o **Glomerular dz in > 80% NA and Eu captive cheetahs on necropsy.** Serum levels of creatinine and urea are not sensitive to early renal damage. Endogenous creatinine clearance previously measured in cheetahs under anesthesia is considered reliable estimate of GFR and early detector of CKD. o Limitations including need for serial, precisely timed blood draws under anesthesia and variability in normal reference range. SDMA exclusively eliminated by glomerular filtration.  Increases as GFR decreases with age in cats.  Performed on whole blood, serum, and plasma.  Stable molecule that remains unchanged even after holding samples for over 7 days at both 4 deg C and room temp.  Correlates well with GFR and serum creatinine in dogs with progressive renal dz. ** Increases detected 6-40 months before increases in urea and creatinine.**

Answer 54

Waugh, L., Lyon, S., Cole, G. A., D'Agostino, J., Cross, J., Strong-Townsend, M., ... & Brandão, J. (2018). **Retrospective analysis and validation of serum symmetric dimethylarginine (SDMA) concentrations in cheetahs (Acinonyx jubatus).** Journal of Zoo and Wildlife Medicine, 49(3), 623-631. Abstract: Kidney disease is common among captive cheetahs (Acinonyx jubatus). Serum creatinine is the most common measurement to estimate glomerular filtration rate (GFR) because of the ease of its clinical use, but it is a crude estimate that only increases after significant disease is already present and is affected by extrarenal factors. Symmetric dimethylarginine (SDMA) is a renal biomarker in humans, dogs, and cats that correlates with serum creatinine and GFR and appears to be an earlier and more specific biomarker for kidney disease. Ninety-two banked serum samples from 11 cheetahs housed at the Oklahoma City Zoo from 1992 to 2012 were retrospectively analyzed. Histopathology results were available for 10/11 cheetahs, and all 10 had histologic renal lesions. **General categories of renal lesions included glomerulosclerosis (7/10; 70%), amyloidosis (7/10; 70%), inflammatory (9/10; 90%), and oxalate nephrosis (2/10; 20%).** SDMA immunoassay and mass spectrometry were measured for validation and compared with creatinine to assess for correlation. Serum creatinine concentrations were determined by enzymatic colorimetric methods. **SDMA immunoassay was validated in cheetahs and correlated well with serum creatinine (R21⁄40.687; P , 0.0001). SDMA and serum creatinine measured from freeze-thawed stored samples show high correlation in individual cheetahs (R2 1⁄4 0.972; P , 0.0001).** These data support that SDMA could be a promising renal biomarker in cheetahs. Further research is warranted to investigate whether SDMA might be an earlier indicator of kidney disease in cheetahs and whether this assay can be extended to other nondomestic carnivores. Key Points: * BUN and Cr requires 75% nonfunctioning nephrons and affected by extrarenal factors (body mass, age) in dogs * SDMA an early indicator of kidney disease that is unaffected by extrarenal factors * Banked serum 11 cheetahs used, all had kidney disease on necropsy (glomerulosclerosis and renal amyloidosis) * SDMA immunoassay was highly correlated with SDMA measured by liquid chromatography-mass spectrometry 🡪 can use to evaluate SDMA in cheetahs * Creatinine and SDMA positively correlated o SDMA predicted reduction in GFR 9.8mo earlier than creatinine * Could not establish reference intervals (all had kidney disease) Take home: SDMA is valid in cheetahs and promising tool for diagnosing kidney disease

Answer 55

Rodriguez, K. T., Gompf, R. E., Smith, C. K., Price, J. M., & Cushing, A. C. (2018). Echocardiographic parameters in eight adult tigers (panthera tigris) during two phases of an anesthetic protocol. Journal of Zoo and Wildlife Medicine, 49(4), 875-880. Abstract: Eight adult tigers (Panthera tigris) underwent a complete echocardiographic examination following sedation with medetomidine, midazolam, and induction of general anesthesia using ketamine and isoﬂurane (phase 1). Atipamezole was used to antagonize medetomidine (phase 2) and a second echocardiographic examination was performed. Physiologic tricuspid and pulmonic regurgitations were common ﬁndings in the sample population and one tiger was excluded from ﬁnal analyses due to the ﬁnding of a ventricular septal defect. Measurements and mean arterial pressure were assessed for statistically signiﬁcant differences between the two examination phases as well as gender and weight. **There was a statistically signiﬁcant difference between interventricular septum thickness at end systole, ejection fraction, and mean arterial pressure between anesthetic phases while fractional shortening and left ventricular internal dimension at end-systole approached, but did not reach, statistical signiﬁcance between phases.** Weight was found to be a statistically signiﬁcant predictor of stroke volume and left ventricular internal dimension at end-diastole. The echocardiographic measurements obtained during this study can be used as guidelines for future examinations in adult tigers. The effects of medetomidine on these measurements and systolic function should be taken into account when performing echocardiograms and monitoring anesthetic events. * Tricuspid and pulmonic regurgitation were common finding (86%) and 100%) o TR - similar to dogs, more than cheetahs (44%) and leopards (33%) * MAP higher in phase 1 – vasoconstriction from medetomidine Take Home: Alpha 2 agonists affect heart measurements during echo – consider reversal

Answer 56

Miller, M. A., Buss, P., Sylvester, T. T., Lyashchenko, K. P., deKlerk-Lorist, L. M., Bengis, R., ... & van Helden, P. (2019). **Mycobacterium bovis in free-ranging lions (panthera leo)—evaluation of serological and tuberculin skin tests for detection of infection and disease. ** Journal of Zoo and Wildlife Medicine, 50(1), 7-15. Abstract: Bovine tuberculosis (bTB), caused by Mycobacterium bovis infection, causes morbidity and mortality in free-ranging lions in bTB-endemic areas of South Africa. However, the only currently used diagnostic test is the tuberculin skin test (TST). This test is logistically challenging to perform because it requires immobilization of lions twice in a 72-hr period. **Blood-based diagnostic tests, such as serological assays, have been previously reported for M. bovis detection in lion populations, and have the advantage of only requiring a single immobilization.** In addition, serological assays can be used for retrospective testing. Therefore, the aim of this study was to test free-ranging lions with the STAT-PAKt (Chembio Diagnostics Systems, Medford, NY 11763, USA) and DPPt VetTB (Chembio Diagnostics Systems) serological assays and compare those results with the tuberculin skin test. The serological assays were also used to determine prevalence in bTB-endemic and uninfected lion populations. **The results showed that the serological assays could distinguish between M. bovis culture-positive and -negative lions. In addition, antigen-specific humoral responses were present in lions that had clinical signs of bTB disease or were shedding M. bovis antemortem.** Although the seroprevalence of M. bovis infection in Kruger National Park lions was similar to that obtained from antemortem mycobacterial culture (4.8 and 3.3%, respectively), it was less than that estimated by the TST (72%). **These findings support the hypothesis that assays based on cell-mediated immune responses are more sensitive than serology is in detecting M. bovis infection in lions. However, serological assays can have a role in bTB disease detection in lions and are especially useful for retrospective studies.** Intro * Bovine TB (Mycobacterium bovis) in South African buffalo populations and other species has resulting in infection and mortality in resident lion prides. * Clinical disease – pulmonary and bone lesions, emaciation and death. * Techniques for detecting M. bovis infection and dz in lions: * Tuberculin skin test – only available antemortem test, commonly used. * Logistically challenging in free-ranging lions. * Cytokine release and gene expression assays recently described in lions, but requires fresh samples and transport. * Preliminary results with serologic tests suggest an association between humoral responses, positive culture results, and presence of lesions in lions. Results: * 52% KNP lions confirmed as infected by culture. * Most were seropositive by STAT-PAK and DPP VetTB, TST reactive. * 8/11 with gross disease on necropsy were reactive with STAT-PAK. * Calculated sensitivities/spec: * STAT-PAK 62.5% sens; 100% spec. * TST 72.7% sens; 80% spec. Key points: * Detection of antibodies may be useful in a population with unsuspected exposure. * STAT-PAK and DPP assays may have greater value in detecting lions with chronic or advanced bTB. * Estimated prevalence in KNP lions 72% based on TST. * Assays based on cell-mediated immune responses more sensitive vs serology for infection. **Takeaway: Serologic tests for bTB can be used in free-ranging lions, main advantage vs the skin test is a single immobilization.**

Answer 57

Broughton, H. M., Govender, D., Shikwambana, P., Chappell, P., & Jolles, A. (2017). **Bridging gaps between zoo and wildlife medicine: establishing reference intervals for free-ranging african lions (panthera leo). ** Journal of Zoo and Wildlife Medicine, 48(2), 298-311. Abstract: The International Species Information System has set forth an extensive database of reference intervals for zoologic species, allowing veterinarians and game park officials to distinguish normal health parameters from underlying disease processes in captive wildlife. However, several recent studies comparing reference values from captive and free-ranging animals have found significant variation between populations, necessitating the development of separate reference intervals in free-ranging wildlife to aid in the interpretation of health data. Thus, this study characterizes reference intervals for six biochemical analytes, eleven hematologic or immune parameters, and three hormones using samples from 219 free-ranging African lions (Panthera leo) captured in Kruger National Park, South Africa. Using the original sample population, exclusion criteria based on physical examination were applied to yield a final reference population of 52 clinically normal lions. Reference intervals were then generated via 90% confidence intervals on log-transformed data using parametric bootstrapping techniques. In addition to the generation of reference intervals, linear mixed-effect models and generalized linear mixed-effect models were used to model associations of each focal parameter with the following independent variables: age, sex, and body condition score. **Age and sex were statistically significant drivers for changes in hepatic enzymes, renal values, hematologic parameters, and leptin, a hormone related to body fat stores**. Body condition was positively correlated with changes in monocyte counts. Given the large variation in reference values taken from captive versus free-ranging lions, it is our hope that this study will serve as a baseline for future clinical evaluations and biomedical research targeting free-ranging African lions. * Variability between captive reference ranges and free-ranging reference ranges is common * Age associated with decreases in WBC and ALP and increases in BUN * Males – higher PCV and leptin, lower ALT and BUN than in females * Monocytes increase with body condition * Higher BUN in wild population o Wild populations drink once a week and have very high protein diets and protracted periods of starvation Take Home: Wild lion parameters more narrow than Species 360 one – associations with age and sex

Answer 58

Schlanser, J. R., Harrison, T. M., Wise, A., & Kiupel, M. (2019). **Differentiating enteropathy-associated t-cell lymphoma type 2 from inflammatory bowel disease in a snow leopard (uncia uncia). ** Journal of Zoo and Wildlife Medicine, 50(2), 474-477. Abstract: After a history of intermittent vomiting, endoscopic biopsies of stomach and duodenum were collected from a 13-yr-old male snow leopard (Uncia uncia). On microscopic examination, monomorphic small lymphocytes expanded the duodenal mucosa and occasionally formed intraepithelial nests. Immunohistochemistry of the infiltrating small lymphocytes in the mucosa and within the epithelium had strong, perimembranous labeling for CD3e, with few CD79a-positive lymphocytes located at the base of the villi. **Polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) of feline T-cell receptor gamma (TCRG) detected a monoclonal cell population. The sequence of the PCR product was 100% homologous with the feline TCRG gene. By histology, immunophenotyping, and PARR testing, a final diagnosis of enteropathy-associated T-cell lymphoma, small cell type, was made.** Homology in the nucleotide sequence between U. uncia and the domestic cat (Felis catus) indicates that feline PARR testing for TCRG may be diagnostic in snow leopards. 13 year old male snow leopard (see abstract) Treatment in this case: chlorambucil (9 mg/kg q48 hrs), Vitamin B, prednisone (0.5 mg/kg q48 hrs). Initial response to clinical signs, but euthanasia performed after 9 months due to returns of clinical signs and repeat biopsies showing persistence of lymphoma in the same locations T-cell lymphoma = lymphoid invasion into the submucosa, tunica muscularis, serosa * Enteropathy-associated T-cell lymphoma (EATL) type 2- usually T-cell restricted to the mucosa * Can be very difficult to differentiate from IBD with just histology (both diseases present as small lymphocytes infiltrating the intestinal villa) * Immunophenotyping- CD3e (T cells) and CD79a (B cells) has been used in some exotic felids for lymphoma diagnosis * **PARR testing on TCRG (for T-cell clonality) used in addition to histology * PARR had not been previously described in large cats ** EATL type 2 diagnosed in this case- most common form of lymphoma in cats (prevalence of up to 70%) Most previously reported cases of T-cell lymphoma in large cats were characterized as multicentric large cell type → intestinal T cell lymphoma has not been commonly reported * Possible that EATL has been underdiagnosed due to diagnosing based on histology alone * Use of PARR testing may improve diagnostic accuracy * CD3e-positive T cells should raise concern for EATL type 2, PARR testing should be then performed

Answer 59

Metrione, L. C., Sim, R. R., Cavanaugh, B., Coty, T., & McCain, S. (2019). **Novel pregnancy diagnosis and altrenogest supplementation lead to successful pregnancy in a black-footed cat (felis nigripes). ** Journal of Zoo and Wildlife Medicine, 50(2), 508-510. Abstract: This is the first time fecal prostaglandin F2a metabolite (PGFM) analysis and altrenogest were used to determine and maintain a pregnancy in a zoo-housed black-footed cat (BFC; Felis nigripes). The established pair had not produced offspring during the year since their arrival at Birmingham Zoo. Fecal samples were collected daily and analyzed by enzyme immunoassay for progestagen, estrogen metabolite, and PGFM concentrations. After a 4-mo separation, the pair was reintroduced, and breeding was reinitiated. Two pregnancies were suggested by sustained, elevated progestagen concentrations (mean .3 3 baseline) but were confirmed by elevated PGFM concentrations (mean 4–25 3 baseline) beginning about 1 mo after presumed or observed breeding. The first pregnancy was lost after ;51 days, and altrenogest was administered (0.088 mg/kg/day) in order to help sustain the second pregnancy, which went to term, but the kitten did not survive. Ultrasonographic and radiographic evaluations of pregnancy were utilized during the second pregnancy to provide information on litter size. Black-footed cat endemic to grasslands of southern Africa; vulnerable IUCN * Not seasonal breeders, will ovulate spontaneously * Nonpregnant luteal phase (13.2 +/- 1.1d) * Full gestation (63-68d) * Elevated fecal estrogen followed by sustained elevated progestagen confirmed estrus followed by ovulation - progesterone 3x above baseline for 51 days * Fecal PGF2a metabolite (PGFM) distinguishes pregnancy from pseudopregnancy in other felids * Increased 10-350x above baseline in pregnant only (not pseudopregnancy) This female - above baseline 28d after initial progestagen increase and remained 4x higher for 24d * PGFM and progestagen both declined to baseline within a day of each other with no evidence of a kitten - dam lost the pregnancy 2nd breeding - viable fetus seen on ultrasound 30 d later * Treated with altrenogest (progestin) 0.088 mg/kg/d starting d31 to sustain pregnancy * PGFM increased 5x above baseline 32d post-breeding then 25x 48-71d * Weaned off altrenogest over 5 d to be fully off by d60 * Parturition d72 * Kitten necropsy: lymphoid follicular depletion on histo, no etiology identified * Possible exogenous progesterone supplementation started too late? Takeaways * First known use of altrenogest to supplement pregnancy of black footed cat * Validation of PGFM assay for pregnancy diagnosis in black footed cat * Breeding and ovulation stimulated by reintroduction after 4 mo of black footed cat pair * Ultrasound and rads useful as ancillary diagnostics to evaluate pregnancy

Answer 60

Hespel, A. M., Leon, J., & Duval, A. (2019). **Radiologic measurement and determination of kidney-to-vertebra ratios as a method of estimating renal size in servals (leptailurus serval). ** Journal of Zoo and Wildlife Medicine, 50(3), 589-593. Abstract: Numerous conditions, from acute to chronic renal changes, have the potential to alter the renal size on radiographs. However, current literature does not provide any radiographic reference range for the normal renal size in the serval (Leptailurus serval). This retrospective study includes 13 healthy adult servals, seen between 2004 and 2017 at the UTCVM Veterinary Medical Center, deemed healthy based on physical examination and blood work. Orthogonal radiographs were anonymized, calibrated, and evaluated blindly by three observers of different experience levels. Renal and individual lumbar vertebral body lengths were measured on both orthogonal views, and ratios of kidney length to each lumbar vertebral length were calculated on both views. On the ventrodorsal projection, the kidney length to L5 vertebra ratio had the lowest standard deviation (SD) with a mean of 1.82, SD of 0.27, and 95% confidence interval (CI) (1.63–2.00). On the lateral view, the kidney length to L3 vertebra ratio had the lowest SD with a mean of 2.03, SD of 0.25, and 95% CI(1.86–2.20). Statistically, all three observers were shown to be in agreement, indicating reproducibility of the measurements among observers with different levels of expertise. * Serval’s native to Africa ; savannahs and streams o Only member of the genus Leptailurus o Nocturnal and solitary. Most commonly owned non-domestic felid. * Suffer from similar diseases to the domestic cat including CKD which is a significant cause of morbidity and mortality in geriatric cats. * Rads are commonly used and noninvasive and may provide info on renal size, shape, opacity and margination which may vary with disease processes. * GOAL - determine normal renal radiographic reference range and ratio for renal length. Results/discussion: * Average renal length 6.61+/-0.96 cm * Av renal length is about 2x L3 on lateral and 1.8x L5 on VD o Note that in domestic cats you kidney should be 2.4-3x length of L2  L3 and L5 were used because they had the lowest SD  Serval vertebra are longer than domestic cats * Renal disease has been reported in panthera, Acinonyx, and Felis * Urolithiasis is amount the most common with Ca (Ca ox and Ca phos) most common followed by struvites * Servals reportedly suffer from cystinuria (27% prevalence) * Can cause urinary obstruction which in domestic cats can result in increased renal size * CKD is very common and often reported to be secondary to amyloidosis, glomerulosclerosis, and pyelonephritis. Prognosis is poor. * In domestic cats CKD commonly associated with decreased renal size TAKE HOME: Normal radiographic renal length 2x L3 on lateral and 1.8x L5 on VD. Renal disease is common in large cats. CKD, often due to amyloidosis, glomerulosclerosis, and pyelonephritis, is a cause of significant morbidity and mortality. Urolithiasis has been reported in several non domestic felids and most commonly is composed of Ca with struvite being the second most common.

Answer 61

Boucher, C. J., Venter, I. J., Tordiffe, A. S., & Kirberger, R. M. (2019). Intraocular pressure, tear production, and ocular biometry in healthy adult cheetahs (acinonyx jubatus). Journal of Zoo and Wildlife Medicine, 50(3), 634-643. Abstract: Schirmer tear test (STT), intraocular pressure (IOP), and ocular biometry were measured in 58 healthy cheetahs (Acinonyx jubatus) between the ages of 2 and 14 yr in two captive groups. During routine health examination under immobilization, each animal underwent ophthalmic examination including STT, IOP, slit-lamp biomicroscopy, indirect ophthalmoscopy, and ocular ultrasonography. Mean ± SD values for STT and IOP were 13.58 ± 5.29 mm/min and 31.40 ± 5.01 mm Hg, respectively. The time between immobilization and examination significantly influenced both STT and IOP measurements. For IOP measurements, a significant group effect was also demonstrated, possibly due to the different anesthetic drug combinations used. Mean ± SD values for biometry were as follows: globe length (32.73 ± 1.74 mm), lens thickness (7.25 ± 1.09 mm), anterior chamber (8.54 ± 1.21 mm), and posterior segment (16.31 ± 2.06 mm). Surprisingly, all of the biometry measurements differed significantly between the two groups. Establishing STT, IOP, and biometric reference values is important for evaluating the ocular health status of the species. These reference values will assist veterinarians working with cheetahs to perform a full and informed ophthalmic examination. * All immobilized- group 1 with ketamine, medetomidine; second group with telazol, medetomidine then intubated and maintained on isoflurane or propofol IV o Tear production (STT), IOP (using a tonovet on the “d” setting) evaluated immediately as soon as they could get hands on the animal, ocular ultrasonography performed o Significant difference between STT values depending on time after immobilization (decreased with increased time) o Significant difference between left and right eyes with right having a lower value o Pressures differed between the two groups – 32 vs 29 in the propofol group. Discussion: * Immobilization affects measurements of tear production and IOP. o IOP lower in the propofol/supplemental iso group. * Tear film = plays important role in health of the conjunctiva and cornea, maintenance of corneal clarity; deficiency of tear production → corneal and conjunctival lesions * STT reading decreased with time, likely impacted by dust/wind/humidity and side of recumbency o Recumbency position may influence the STT values o Tear production in cheetah lower than reported in domestic cats, dogs, african lions

Answer 62

Girling, Simon J., et al. USE OF CLINDAMYCIN IN PALLAS'CATS [OTOCOLOBUS (FELIS) **MANUL] TO REDUCE JUVENILE TOXOPLASMOSIS-ASSOCIATED MORTALITY RATES.** Journal of Zoo and Wildlife Medicine 51.1 (2020): 39-45. Abstract: Pallas' cat [Otocolobus (Felis) manul] experiences a high mortality rate from toxoplasmosis. During the period 2006–2016, the overall mortality rate for this species from all causes during the first year of life was 71.59% in European Association of Zoos and Aquaria institutions, with the most significant infectious cause from systemic toxoplasmosis (20.6%) as confirmed by postmortem examination and histopathology. Clindamycin was used starting in 2014 in two collections that had previously experienced 100% mortality rates by toxoplasmosis in kittens less than one year of age, covering key Toxoplasma gondii exposure periods for kittens (n = 17) as a prophylactic measure. This protocol resulted in a 67.03% (95% confidence interval 41.76–78.61%) reduction in the first year mortality rate over a two-year period to 5.88% in those animals treated. Key Points: Toxoplasma gondii. * Felids definitive host, only spp that pass oocysts. * Most felid infections via ingestion of cysts from IM host tissues, or direct ingestion of oocysts. * Adult felids typically subclinical. * Immune system suppresses infection, shedding typically occurs once in the hosts life. * Fatal infection from replication of organism in tissues, resulting in granulomatous inflammation and necrosis. * Lesions most common in lungs, liver, heart, spleen, pancreas, mesenteric LN. Palas cat – Indigenous to central Asia. * Far greater susceptiblility to T. gondii than other felids and chronic seropositivity with ongoing veritical transmission being frequently reported in captive animals. * Low seropositivity < 13% in the wild. * Reported to cause high mortality rates in managed care ~60%. * Kittens around weaning, when exposure is greatest and immune system not fully developed. * Losses also late in gestation or after parturition with infection of kittens postnatally via felid feces, prey items, in utero. * Domestic cats – Lactational transmission reported experimentally, * Once clinical, treatment generally unsuccessful. * Most common infectious dz assoc with mortality in annual mort records of European Assoc Zoo and Aquaria = toxo (20.6%) kittens. * Prophylactic medical management regime with clindamycin formulated and tested at two institutions experiencing high kitten mortality. M+M: * Pregnant females at the two collections that were seropositive for toxo were treated with clindamycin 10-12.5 mg/kg PO q12h x 7-10 days preparturition until 3-4 wks postparturition. * Once eating solid foods, ~3-4wks, kittens administered clindamycin at same dose for 3-4 wk to cover exposure period where previous losses had been reported in both collections. * Clindamycin capsules opened and powder spread onto food items, target fed to individuals. Results: * 2012-2013 period without clindamycin protocol – 9 born, 9 died. * 2014-2016 period with clindamycin protocol – 17 born, 1 died. Discussion: * Toxo considered primary pathogen in Pallas’ cats because of their evolutionarily naïve immune systems. * **Other dz that impact Pallas cats – Trypanosoma manulis, Cytauxzoon felis, Mycoplasma sp, FHV-1, FIV, FeLV, influenza A.** Periods of high exposure risk for toxo: * Late term gestation and around weaning when maternal immunity is waning, permanent immunity has not yet fully developed, and kittens are exposed to additional sources of infection i.e. consumption of wild rodents. * Pallas cats also known to die beacause of either immunosuppression or failure to develop immunity through lack of exposure in early life. Historically used combo tx with sulfadiazine and pyrimethamine for tx of domestic felids. * Pyrimethamine may exacerbate negative effects of sulfonamide drugs, leads to anorexia, leukopenia, anemia, hypersalivation. * Pyrimethamine alone assoc with myelosuppression in felids, contraindicated. * Diclazuril also suggested as preventative therapy for toxo in Pallas cats. * Combination with clindamycin may have better success. * Formulation of diclazuril in Eu predominantly sold as in-feed mix for poultry and farmed rabbits. Hard to titrate dose for kittens. Clindamycin = tx of choice for clinical toxoplasmosis in the domestic cat. MOA - 50s ribosome * Long term administration associated with dysbiosis, vomiting, diarrhea, predisposition to Clostridium spp and yeast in GIT. * Generally well tolerated unless liver or kidney dz in domestic cats. * Clindamycin hindered recovery for a single case of ocular toxo in domestic felids, possibly by inhibition of natural phagocyte function. Other Treatment Options * TMS has also been used as a prophylactic or preemptive therapy but serious side effects in felids. * Ponazuril shown to reduce mortality rates of toxo in mice. Also toltrazuril metabolite. * Both have GI side effects. Excreted metabolite may be toxic to plants, survives for long periods in the environment. Management * Reduction of exposure risk – Freezing, defrosting fed foods to reduce viability of bradyzoites present in prey items. * Consider housing animals indoors to avoid hunting wild rodents and infection via that route. * Deaths have occurred in completely indoor-housed cats later moved to outdoor enclosures.

Answer 63

Flaminio, K. P., Christensen, J. M., Alshahrani, S. M., & Mohammed, S. M. (2019). **Pharmacokinetics after subcutaneous administration of a single dose of cefovecin sodium in African lions (Panthera leo). ** American journal of veterinary research, 80(3), 230-234. OBJECTIVE: To determine the pharmacokinetics of cefovecin sodium after SC administration of a single dose to African lions (Panthera leo). ANIMALS: 3 adult (9 to 10 years old; 1 male and 2 females) and 3 juvenile (2 years old; 1 male and 2 females) African lions. PROCEDURES: A crossover study was conducted. A single dose of cefovecin was administered SC at 4 mg/kg (half dose) and 8 mg/kg (full dose) to African lions. Blood samples were collected daily for 14 days after cefovecin injection. Plasma drug concentrations were determined by use of high-performance liquid chromatography with UV detection. RESULTS: **Cefovecin had first-order elimination kinetics for doses of 4 and 8 mg/kg. Mean ± SD maximum plasma concentration was 9.73 ± 1.01 μg/mL and 18.35 ± 0.94 μg/mL after doses of 4 and 8 mg/kg, respectively. Time to maximum plasma concentration was approximately 4 hours for both doses. Mean elimination half-life was approximately 111 and 115 hours after doses of 4 and 8 mg/kg, respectively.** CONCLUSIONS AND CLINICAL RELEVANCE: Cefovecin was detected in lion plasma for **336 hours after administration at both 4 and 8 mg/kg** at concentrations greater than the reported minimum inhibitory concentration (0.06 μg/mL) for common bacterial organisms in domestic cats. These results indicated that cefovecin administered at 4 mg/kg SC reached therapeutic concentrations for an extended period in African lions. Key Points: Cefovecin sodium lasts 5 days and up to 7 days in dogs and cats, respectively - high protein binding and high bioavailability after SC administration - Protein binding means not consistent across spp, PK/PD studies for each need to be undertaken Dilution to higher concentration caused no adverse effects (better for larger animals) Cmax substantially lower for African lions after administration of 4 and 8 mg/kg compared with the value for tigers, domestic cats, and dogs - Reached the Cmax more quickly in lions (4 hours) than in dogs (6.2 hours) and tigers (22.3 hours) but more slowly than in domestic cats (1 to 2 hours) - Cmax of cefovecin in lions higher than the minimum effective concentration extrapolated from other species for both doses - Reached concentrations that would be therapeutic against bacterial organisms by 4 hours after SC administration - Tigers and lions related phylogenetically but also have different PK results with meloxicam - can’t assume based on genetic relation - Higher AUC and lower clearance = lower dose acceptable (4mg/kg) in lions - 4mg/kg q14d sufficient, also safe to increase to 8mg/kg bc of linear PK Take home points: SC administration of cefovecin to African lions appeared to be safe and resulted in effective plasma concentrations with a pharmacokinetic profile more similar to those of domestic dogs and cats than to those of other exotic species.

Answer 64

Cushing, A. C., Ramsay, E. C., Steeil, J., & Cox, S. (2017). **Pharmacokinetic parameters of cefovecin sodium (Convenia) in captive tigers (Panthera tigris). ** Journal of Zoo and Wildlife Medicine, 48(4), 1188-1192. Abstract: **A single 8 mg/kg dose of Cefovecin (Convenia®) was administered intramuscularly in the hindlimb of eight anesthetized captive tigers (Panthera tigris) **and serial blood samples were collected over the next 56 days to determine pharmacokinetic characteristics. High-performance liquid chromatography was utilized to determine plasma levels, and demonstrated a mean terminal half-life of 227.8 ± 29.3 hr. Cefovecin, >1 μg/ml, was detected in plasma levels up to 56 days post injection. Protein binding of tiger plasma for cefovecin was 98%. The long duration of therapeutic plasma concentrations and lack of adverse effects make cefovecin a useful antimicrobial drug for tigers. Key Points: - Cefovecin sodium (Convenia) – 3rd generation, long-acting injectable cephalosporin. -- In dogs, long elimination half-life 5.5 days, cats 7 days. -- Recommended dose 8 mg/kg. - No adverse effects noted. - Highest conc at 90 min postadministration. - First decline noted 2 days later. - Absorption slower than in domestic cats. - Plasma concentrations maintained for at least 49 days.

Answer 65

Whitehouse-Tedd, K. M., Smith, L., Budd, J. A., & Lloyd, C. G. (2017). **Suspected adverse reactions to oral administration of a praziquantel-pyrantel combination in captive cheetahs (Acinonyx jubatus). ** Journal of the American Veterinary Medical Association, 251(10), 1188-1195. Abstract: OBJECTIVE To characterize adverse reactions to oral administration of a combination of praziquantel and pyrantel embonate or pyrantel pamoate, with or without oxantel embonate, in captive cheetahs (Acinonyx jubatus). DESIGN Retrospective case series and case-control study. ANIMALS 16 captive cheetahs with signs of adverse reaction to oral administration of praziquantel and pyrantel, with or without oxantel embonate (affected group), and 27 cheetahs without such reactions (unaffected group), all from 3 independent facilities. PROCEDURES Medical records and postmortem findings for affected cheetahs were reviewed and compared with those of unaffected animals. Anthelmintic doses administered, age, and sex of cheetahs were compared between groups. RESULTS 3 reactions in affected cheetahs were fatal, whereas the remainder ranged from mild to severe. Postmortem examination failed to reveal any disease processes or conditions to explain the deaths. No differences in anthelmintic dose were identified between affected and unaffected cheetahs for all facilities combined, and no correlation existed between dose and reaction severity. No association with sex was detected, but affected cheetahs were significantly younger than unaffected cheetahs. This difference was not significant after controlling for facility. CONCLUSIONS AND CLINICAL RELEVANCE **Cheetahs were concluded to have had an adverse reaction to the praziquantel-pyrantel combination because of temporal proximity of onset of clinical signs to dose administration, similarity of signs to those reported for toxicosis in other species for these drugs, and a lack of other disease process or environmental explanatory factors. A highly cautious approach to the use of this drug combination is recommended for cheetahs.** Key Points: - Signs in these cheetahs suggestive of toxicity -- Within 4 hours of administration -- Neurologic – ataxia, seizures, eventually pulmonary hemorrhage & respiratory distress -- No consistent drug overdose document -- Cheetahs with fatlity had evidence of gastritis – may have enhanced absorption of these drugs - Praziquantel -- Toxicity infrequently reported – weakness, vomiting, depression, ataxia - Pyrantel -- Poor GI absorption due to salt (pamoate or embonate) -- Tachycardia, hypersalivation, diarrhea, vomiting, tremors, convulsions, excitation, ataxia Take-Home: Gastritis may enhance absorption of drugs which could predispose to adverse events. Careful with praziquantel and pyrantel administration.

Answer 66

Newkirk, K. M., Beard, L. K., Sun, X., & Ramsay, E. C. (2017). **Investigation of enrofloxacin-associated retinal toxicity in nondomestic felids. ** Journal of Zoo and Wildlife Medicine, 48(2), 518-520. Abstract: Enrofloxacin is known to cause retinal toxicity in domestic cats. The hallmark lesion of enrofloxacin-associated retinal toxicity in domestic cats is thinning of the outer nuclear layer of the retina. Enrofloxacin is commonly used to treat bacterial infections in nondomestic felids because of its action against a wide spectrum of bacteria and the ability for it to be given orally. No previous studies have investigated the potential retinal toxicity of enrofloxacin in nondomestic felids. **This retrospective study evaluated 81 eyes from 14 lions (Panthera leo) and 33 tigers (Panthera tigris) that had been enucleated or collected postmortem. **The thickness of the outer nuclear retina was assessed in two separate sites in each eye by counting the rows of nuclei and by using digital image analysis software to determine the area of the nuclei at each site. Medical records were reviewed to determine the enrofloxacin dose for each cat. **Cats that had not received enrofloxacin (n = 11) were compared with treated animals (n = 36). The outer nuclear layer thickness or area in treated versus untreated cats was not significantly different. Additionally, no clinical blindness was reported in any of the cats. This study showed no evidence of enrofloxacin-associated thinning of the outer nuclear layer in the lions and tigers evaluated, suggesting that enrofloxacin can be used safely in these animals.** Key Points: - Domestic cats – enro assoc with acute blindness 2-12 days after oral administration. -- Affects photoreceptor outer segments (rods and cones). -- Tapetal hyperreflectivity and attenuation of retinal vessels on fundic exam. -- Thinning of outer nuclear layer. Inner nuclear layer and ganglion cell layers unaffected. - UT tigers and lions – enro dose 5 mg/kg PO q24h commonly used. Necropsy review. -- No clinical blindness reported in any cats. -- No evidence of enro-associated thinning of the outer nuclear layer in any cats. - Mechanism of this toxicity not fully understood. ABCG2 transporter gene expressed in retinal capillary endothelial cells, plays a role in maintaining blood-retinal barrier and protecting the retina from phototoxic compounds. Also involved in transporting enro in the body, may play role. - Administration of enro to nondomestic felids within the dose recommended for oral administration in domestic cats for 14 days is not associated with thinning of the outer nuclear layer.

Answer 67

Huckins, G. L., Chinnadurai, S. K., Ivančić, M., Bergmann, J., Balko, J. A., Aitken-Palmer, C., ... & Cook, J. L. (2018). **Osteochondral autograft transfer for treatment of stifle osteochondritis dissecans in two related snow leopards (panthera uncia). ** Journal of Zoo and Wildlife Medicine, 49(3), 788-793. Two related female snow leopards (Panthera uncia) were diagnosed with **osteochondritis dissecans of the lateral femoral condyle via radiographs and computed tomography**. Lesions were either bilateral (Patient 1) or unilateral (Patient 2). **Surgical treatment was performed using an osteochondral autograft transfer system following cartilage flap removal.** After a 6-wk recovery period, both animals had a markedly improved gait with mild pelvic limb stiffness. Patient 1 developed intermittent lameness 9 mo postsurgery and Patient 2 had mild lameness 3 yr postsurgery. Because of recurrent lameness, both animals were treated with platelet-rich plasma intra-articular injections and oral gabapentin (Patient 1) or oral meloxicam (Patient 2). Full incorporation of autografts was confirmed with computed tomography (both patients) and arthroscopy (Patient 1 only). Osteochondritis dissecans has been previously reported in snow leopards and these cases represent the first reported use of osteochondral autograft repair and platelet-rich plasma to treat the condition. Key Points: - Osteochondrosis – dysfunctional endochondral ossification; thickening and necrosis of cartilage, fissures, formation of cartilaginous flap. -- OCD when flap detaches from underlying subchondral bone. -- Multifactorial – trauma, nutrition, hormones, blood supply, genetics. -- OCD in large felids has been tx with cartilage flap removal and debridement. - This report – snow leopards treated for OCD with osteochondral autograft transfer. -- Osteochondral Autograft Transfer System. -- Osteochondral autograft removed from lateral femoral sulcus terminalis and proximal aspect of the trochlear ridge, trimmed to size and inserted into recipient socket. -- Both patients did well, some intermittent lameness, both also tx with PRP. Takeaway: An osteochondral autograft transfer procedure may provide effective tx of stifle OCD lesions in snow leopards.

Answer 68

Knollinger, A. M., McDonald, J. E., Carpenter, N. A., & Crook, E. K. (2018). **Use of equine amniotic membrane free-island grafts for treatment of a midstromal corneal ulcer and descemetocele in a snow leopard (Panthera uncia). ** Journal of the American Veterinary Medical Association, 253(12), 1623-1629. CASE DESCRIPTION A 7-year-old sexually intact female snow leopard (Panthera uncia) was examined because of blepharospasm, periocular discharge, ventral deviation of the upper eyelid cilia, third eyelid prolapse, and corneal opacity of the right eye. CLINICAL FINDINGS An ophthalmic examination performed with the patient anesthetized revealed a 3 X 3-mm ulcer that extended approximately 60% of the depth of the right cornea and was accompanied by perilesional and intralesional cellular infiltrates and active vascularization. The upper eyelid of the right eye also had a previously repaired coloboma resulting in trichiasis. TREATMENT AND OUTCOME Surgical intervention was elected after 5 weeks of medical management including topical administration of autologous serum and topical, subconjunctival, and systemic administration of antimicrobials failed to yield any improvement in the ulcer. Equine amniotic membrane free-island graft placement and eyelid revision surgeries were performed. Two and a half weeks later, a descemetocele was diagnosed ventrolateral to the original ulcer, and a second equine amniotic membrane free-island grafting procedure was performed. Both grafts healed without further intervention. CLINICAL RELEVANCE Equine amniotic membrane free-island grafts were used to successfully repair a corneal ulcer and descemetocele in a snow leopard. The grafting procedure spared the affected globe and resulted in satisfactory cosmesis and functional vision. This procedure should be considered as an option for corneal repair in nondomestic species for which postoperative care and medical treatment options are limited Key Points: - 7-year-old FI snow leopard developed a corneal ulcer -- No improvement with 5 weeks of medical management -- Surgery - midstromal keratectomy and equine amniotic membrane free-island graft - 2.5 wk later – focal dehiscence with descemetocele with prominent perilesional cellular infiltrate and keratomalacia, secondary infection - second equine amniotic membrane free-island graft placed - Free-island graft -- provides tectonic support within corneal defect without use of a pedicle -- removes risk for graft retraction caused by a pedicle attachment -- disadvantage - lack of active vascularization directly to site of stromal loss - Amniotic membrane -- anti-immunogenic, anti-angiogenic, and antimicrobial properties -- ability to facilitate migration, differentiation, and adhesion of corneal epithelial cells; modulate corneal stromal scarring; and decrease corneal surface inflammation -- disadvantages - extensive and complicated procedures required for harvesting and preparing tissue, need to maintain tissue in appropriate ultracold storage (–80°C)

Answer 69

**Unusual splenic B-cell lymphoma in two related Sumatran tigers (Panthera tigris sumatrae)** JAVMA 2020;257:1288–1293 CASE DESCRIPTION A 14-year-old 120-kg (264-lb) sexually intact male Sumatran tiger (Panthera tigris sumatrae) and its 10-year-old 130-kg (286-lb) sexually intact male offspring were housed separately and evaluated independently after experiencing weeks of ongoing malaise, weight loss, and anorexia. CLINICAL FINDINGS Both animals were immobilized and anesthetized for physical examinations and diagnostic testing. Complete blood counts revealed leukopenia and anemia in both tigers. Splenomegaly was identified on abdominal ultrasonography. **Cytologic examination and immunohistochemical staining of splenic samples confirmed intermediate to large B-cell lymphoma; no evidence of lymphoma in surrounding organs was noted. ** TREATMENT AND OUTCOME The sire was treated with lomustine and prednisolone. This tiger was euthanized 21 months after initiation of treatment because of chronic progressive renal disease. The male offspring was treated with l-asparaginase but did not respond to the treatment. A splenectomy was performed, and malaise and anorexia resolved. No further chemotherapy was administered, and the male offspring was instead maintained on a low dose of prednisolone. Thirty-two months after diagnosis, the male offspring was still considered to be in remission. CLINICAL RELEVANCE **To our knowledge, this was the first known report of the diagnosis and management of a splenic B-cell lymphoma in a tiger.** Both tigers achieved positive clinical responses and long-term survival by means of different treatment modalities. The finding of such an unusual neoplasm in a male tiger and its male offspring was noteworthy, raising the possibility of a genetic predisposition for this lymphoma type Case report - 14 yo MI Sumatran tiger (Panthera tigris sumatrae) and his 10-year-old MI offspring were housed separately and evaluated independently after experiencing weeks of ongoing malaise, weight loss, and anorexia. - No abnormalities on PE - CBC revealed anemia and leukopenia in both animals - AUS showed splenomegaly and cytology with IHC showed intermediate to large B cell lymphoma with no evidence of mets to surrounding organs - The sire was treated with lomustine PO q 6 weeks and pred 1mg/kg PO SID - AUS 4 months later revealed a reduction in the size of the spleen and an improved PCV - During the next year the tiger developed hydronephrosis secondary to ureteroliths and was eventually euthanized due to renal failure, however lymphoma was confirmed in the kidneys, mesenteric LN’s and spleen on necropsy - The offspring presented for similar clinical signs 4 years after the sire’s initial presentation - Treated with L-spar and pred (2mg/kg PO SID) - 2 months later, still lethargic and hyporexic, decided to do a splenectomy - Marked clinical improvement within 1 week - Still in remission 32 months later Discussion - **Reports on malignant lymphomas in exotic felids indicate primarily a T-cell origin, few reports of B cell** - Both tigers had splenic marginal zone lymphoma - Both tigers were negative for FeLV and FIV - Possible genetic predisposition due to relatedness of the tigers - 2 different treatments, lomustine showed clinical improvement but disease still present on necropsy and may have contributed to renal failure. L spar + pred + splenectomy yielded a long remission

Answer 70

Yong, J. A. A., Lewis, D. D., Citino, S. B., Cunningham, M. W., Cross, A. R., Farese, J. P., & Pablo, L. S. (2018). **Surgical management of appendicular long-bone fractures in free-ranging florida panthers (puma concolor coryi): six cases (2000–2014). ** Journal of zoo and wildlife medicine, 49(1), 162-171. Abstract: The clinical outcomes of six free-ranging Florida panthers (Puma concolor coryi) that underwent surgical stabilization of appendicular long-bone fractures (three femoral fractures, one tibial and one tibial and fibular fracture and two radial and ulnar fractures) were evaluated. These panthers presented to the University of Florida from 2000-2014. Estimated age of the panthers ranged from 0.5 to 4.5 yr, and weights ranged from 22 to 65 kg. Causes of injuries were vehicular collision (n = 4) and capture related ( n = 2). All panthers underwent open reduction and fracture stabilization. Fixation failure necessitated three subsequent surgeries in one panther. Five panthers survived the immediate postoperative period, and all of these panthers' fractures obtained radiographic union (range, 8-36 [mean, 22] wk). The five surviving panthers underwent convalescence for 7-14 mo at White Oak Conservation Center before being released back into the wild; however, one panther was killed when hit by a car 3 days after release. The remaining four panthers were tracked for up to 106 mo in the wild and successfully integrated back into the native population. **Surgical stabilization of appendicular long-bone fractures in free-ranging Florida panthers can be successful, but must take into account the stress that a large, undomesticated felid will place on the stabilized limb during convalescence as well as the difficulties involved in rehabilitating a wild panther in captivity.** Key Points: - 6 panthers – 3 femoral fx, 1 tibial and fibular fx, 2 r/u fx. - Repaired with variety of internal fixation methods – plates, ILN, cerclage, ex fix. - Time to radiographic union in surviving individuals 8-36 wks. - Open, contaminated fx site shown to be a high predictive risk factor for infection and nonunion. - One panther required several surgical revisions, had catastrophic failure of fixation. -- Massive mechanical demands imposed on implants. -- Locking plate thought to provide improved stability, allowed limited disruption of developing callus during surgical revision. **Takeaway: Robust fixation recommended when stabilizing appendicular long-bone fxs in large felids.**

Answer 71

Ayala, I., Laredo, F., & Alberca, F. (2018). **Benign idiopathic esophageal stricture in a lion (panthera leo): dilation by an achalasia balloon.** Journal of Zoo and Wildlife Medicine, 49(1), 193-195. Abstract: A 1-yr old female lion (Panthera leo) was referred with a 10-mo history of dysphagia for solid food (meat), episodic regurgitation, and poor weight gain. Esophagoscopy confirmed an esophagitis (midesophagus) and a stricture estimated to be of 13 mm diameter. This was subsequently dilated using a 20-mm-diameter balloon for 2 min followed by a 35-mm achalasia balloon for 3 min. The etiology remains undetermined in spite of a thorough history. The animal progressed satisfactorily, reaching 124 kg after 1 yr and has had no further signs. To the authors’ knowledge, neither idiopathic esophageal stricture nor dilation using an achalasia balloon has been reported in Panthera spp. * Survey neck and thoracic radiographs did not show any abnormalities * Esophagoscopy confirmed mid esophageal stricture + esophagitis * Balloon dilation: applying progressive radial force when inflating o No complications, no further signs 1 year out * FB trauma (bones), gastroenteritis, drug induced suspected etiologies Take home: Idiopathic esophageal stricture successfully treated with achalasia balloon dilation

Answer 72

Shimamura, S., Shiota, Y., Takagi, N., Habara, T., Hirata, S., Komai, H., ... & Shimada, T. (2017). Ebstein anomaly in the tsushima leopard cat (prionailurus bengalensis euptilurus). Journal of Zoo and Wildlife Medicine, 48(2), 586-589. Abstract: Ebstein anomaly is a rare congenital heart disease that has been described in domestic dogs, a meerkat, a pygmy goat, and a lion. An 11-mo-old Tsushima leopard cat presented to Osaka Prefecture University Veterinary Hospital for diagnosis and treatment of right-sided congestive heart failure. Echocardiography showed a dilated right atrium and ventricle with an enlarged tricuspid valve annulus and apical displacement of the tricuspid valve leaﬂets. The cat was diagnosed with Ebstein anomaly. To the best of our knowledge, this is the ﬁrst report of this type of congenital heart disease in a Tsushima leopard cat. Case of Ebstein anomaly in a Tsushima leopard cat bred in the Kyoto City Zoo - 11-mo-old intact male Tsushima leopard cat - Poor appetite with mild weight loss, exercise intolerance, ascites, and jaundice - CBC/chem - elevated Tbili, ALT, AST, and GGT - Contrast rads – barium aspiration - Treated with furosemide, monoammonium glycyrrhizinate, and ursodeoxycholic ester - Improved appetite but no improvement in ascites or exercise intolerance - Echo a month later showed R atrium and ventricular enlargement 🡪 suspect RCHF - Treated with furosemide, benazepril, pimobendane - 92 days later – referred - grade 4/6 regurgitant systolic murmur ausculted - CBC/chem repeated 🡪 tbili elevated, otherwise WNL - ECG 🡪 mildly increased P wave - Rads – cardiomegaly, no ascites - Echo 🡪 tricuspid valve dysplasia similar to Ebstein anomaly - Pimo dose decreased but meds continued Ebstein anomaly and tricuspid valve dysplasia - Congenital anomaly - Ebstein anomaly - delamination of septal and inferior leaﬂets incomplete - valve oriﬁce lies apically (displaced posteriorly, with an anterior leaﬂet that occasionally obstructs the right ventricular outlet because the point of distal insertion is abnormal) - adherence of these leaﬂets to the underlying myocardium - tricuspid valve dysplasia - valve dysplastic and normally delaminated, may be stenotic and/or incompetent

Answer 73

Rashid, I., Akbar, H., Gharbi, M., Riaz, F., Islam, S., Saleem, M. B., ... & Ashraf, K. (2017). **First report of trypanosoma evansi infection (surra) in a puma (felis concolor) of lahore zoo, pakistan. ** Journal of Zoo and Wildlife Medicine, 48(3), 918-921. Abstract: The blood protozoan Trypanosoma evansi, which is transmitted by biting flies, is frequently neglected due to subclinical infections. This report describes a case of trypanosomiasis due to T. evansi in a 9-yr-old male puma (Felis concolor) housed at the Lahore Zoo in Pakistan. **Early in January 2015, this male puma presented with chronic lethargy, weight loss, incoordination, hyperthermia, anorexia, sunken eyes, and unthriftiness. Microscopic examination of Giemsa-stained blood smears showed numerous Trypanosoma parasites. The puma was treated with diminazene aceturate subcutaneously twice. A few days later, a blood smear examination showed absence of trypanosomes. Five months later the cat presented with acute epistaxis and died. Postmortem examination showed emaciation, pale liver and kidneys, and hemorrhages on the spleen. Examination of a blood smear taken at the time of death showed numerous Trypanosoma parasites. **PCR testing confirmed the presence of Trypanosoma DNA. DNA sequencing of two amplicons confirmed the presence of Trypanosoma in the blood smears with a 98–99% identity with the previously identified GenBank sequences. A phylogenetic tree was then constructed. Further studies are needed to improve our knowledge about the epidemiology and pathogenesis of T. evansi infection in wild animal species. o Trypanosoma evansi is the most widely distributed pathogenic salivarian trypanosome in animals.  Zoonotic, causes surra. Tsetse flies important potential carrier that can spread dz.  Affects mainly camels, but also horses, cattle, sheep, dogs.  Reported in wild Asiatic black bears.  Little or no pathogenic effect in wildlife, but CS may occur when the host is stressed.

Answer 74

Eustace, R., Rubin, J., Thompson, K. A., Snowdon, K., Sikarskie, J. G., Monahan, C., & Smedley, R. C. (2017). **Diagnosis and treatment of a unilateral renal cystadenoma in an african lion (panthera leo). ** Journal of Zoo and Wildlife Medicine, 48(3), 906-909. Abstract: A renal tubular cystadenoma was diagnosed in a 14-yr-old male African lion (Panthera leo). During a routine health evaluation, a left renal mass was identified via physical examination, radiographs, and abdominal ultrasonography. The mass was 30 x 15 cm in size and had a thin capsule with central hypoechoic fluid, suggestive of a perirenal cyst. An exploratory celiotomy with partial nephrectomy was performed without complications. Histologically, the tumor was characterized by a thick fibrous capsule surrounding multiple, variable-sized cysts that markedly compressed the adjacent fibrotic and atrophied renal cortex. Immunohistochemical labeling for Aquaporin-1 and Tamm-Horsfall protein was consistent with a renal tubular cystadenoma of proximal tubule origin. **Renal cystadenomas are an uncommon benign epithelial neoplasm.** There are only two documented case reports in domestic cats. This report represents the first documentation, to the authors’ knowledge, of a renal cystadenoma in a lion. * Abdominal mass found on exam – cystic structure with thick wall found on US * 60 mL of transudate fluid drawn not consistent with uroabdomen * Due to chronic kidney issues, a partial nephrectomy was performed * Seroma occurred at incision site – treated with convenia and it resolved * Renal neoplasms in cats o Lymphoma is the most common o Renal cystadenomas are uncommon (2 cases in domestic cats) * Other differentials – perirenal pseudocysts, PKD, abscesses, hematomas, granulomas

Answer 75

**Carnivore protoparvovirus 1 (parvoviruses) at the domestic–wild carnivore interface in India. ** Shetty, B.D., Zachariah, A., Farver, T.B., Smith, B., Goldstein, T. and Mazet, J.A. Journal of Zoo and Wildlife Medicine, 2020;50(4):1016-1020. Abstract: Carnivore protoparvovirus 1 (CP1, earlier called Feline panleukopenia virus) variants such as canine parvovirus (CPV) and feline parvovirus (FPV) are significant, emerging, multihost pathogens of domestic and wild carnivores. **The diversity of CP1 variants was studied between 2008 and 2014 in Wayanad, India, where flagship wildlife species such as tigers (Panthera tigris) and leopards (Panthera pardus) coexist alongside domestic carnivores, including dogs (Canis lupus familiaris) and cats (Felis catus). Using polymerase chain reaction, FPV and CPV sequences were obtained from the heart blood of a necropsied leopard individual for the first time in the world and from rectal swabs of three sympatric and clinically ill domestic dogs**. CP1 amplicons were also detected in a tiger. Cross-species transmission possibilities were identified, as the closest relatives to the leopard FPV sequence were found in domestic cats from a neighboring state. Carnivore protoparvovirus 1 - previously Feline panleukopenia virus o Single stranded DNA, 4 variants (feline parvovirus, canine, raccoon, and mink enteritis virus) o Cross-species transmission: CPV-2a from domestic dogs to captive large felids (Siberian tiger) o Outbreaks in captive wild carnivores and non-human primates o Leukopenia, fever, depression, dehydration, diarrhea, rarely myocarditis, cerebellar hypoplasia, reproductive losses Positive on PCR: 1/11 tigers, ⅓ leopards, 3/3 domestic dogs, 0/41 large carnivore feces, 0/2 dhole, 0/3 leopard cats o Tiger - kidney o Leopard - heart blood and brain: feline parvovirus o Domestic dogs - rectal swabs: canine parvovirus (100% identical) o Leopard FPV and dog CPV shared 97% genetic identity o Leopard FPV was closest in GenBank to domestic cat FPV amplified in neighboring state **FPV-positive leopard necropsy findings: massive pulmonary hemorrhages interspaced with congested lung nodules with lung flukes (Paragonimous westermani), secondary bacterial infections** Takeaways: * First genetic sequences identified in free-ranging leopards * First report of CP1 PCR amplicons in free-ranging tigers * Genetic proximity of leopard and domestic cat FPV supports multihost dynamics of CP1 variants

Answer 76

**Tick paralysis in a free-ranging bobcat (Lynx rufus)** J Am Vet Med Assoc 2020;256:362–364 CASE DESCRIPTION **A free-ranging male bobcat (Lynx rufus) was evaluated because of signs of pelvic limb paralysis. ** CLINICAL FINDINGS Physical examination of the anesthetized animal revealed tick infestation, normal mentation, and a lack of evidence of traumatic injuries. Radiography revealed no clinically relevant abnormalities. Hematologic analysis results were generally unremarkable, and serologic tests for exposure to feline coronavirus, FeLV, FIV, and Toxoplasma gondii were negative. Results of PCR assays for flea- and common tick-borne organisms other than Bartonella clarridgeiae were negative. TREATMENT AND OUTCOME **Ticks were manually removed, and the patient received supportive care and fipronil treatment. The bobcat made a full recovery within 72 hours after treatment for ticks, and a presumptive diagnosis of tick paralysis was made. Identified tick species included Dermacenter variabilis, Amblyomma americanum, and Ixodes scapularis. ** CLINICAL RELEVANCE To the authors’ knowledge, tick paralysis has not previously been reported in felids outside Australia. This disease should be considered a differential diagnosis in felids, including exotic cats, with signs of neuromuscular disease of unknown etiopathogenesis. Case report ● ~15 month old free ranging MI bobcat (Lynx rufuus) presented for pelvic limb paralysis ● Anesthetized for evaluation, no traumatic injuries found ● Rads and CBC/Chem unremarkable ● FeLV/FIV and toxo negative ● Flea and tick panel negative except bartonella were negative ● 20 ticks were seen and easily reoved from the animal ● Supportive care – methylprednisolone, LRS, ccfa, ivermectin, buprenorphine, and fipronil ● Patient recovered fully after 72 hours – presumed diagnosis of tick parlysis ● Identified tick species included **Dermacenter variabilis**, Amblyomma americanum, and Ixodes scapularis ● First report of tick paralysis in a felid outside of Australia **● Mechanism: The neurotoxin in the salivary glands of the tick blocks the release of transmitters from motor nerve terminals, resulting in a flaccid paralysis that is frequently first observed as hind limb incoordination**

Answer 77

**FELINE CORONAVIRUS AND FELINE INFECTIOUS PERITONITIS IN NONDOMESTIC FELID SPECIES** Journal of Zoo and Wildlife Medicine 52(1): 14–27, 2021 Abstract: Feline coronavirus (FCoV) is reported worldwide and known to cause disease in domestic and nondomestic felid species. Although FCoV often results in mild to inapparent disease, a small subset of cats succumb to the fatal, systemic disease feline infectious peritonitis (FIP). An outbreak of FIP in Cheetahs (Acinonyx jubatus) in a zoological collection demonstrated the devastating effect of FCoV introduction into a naıve group of animals. In addition to cheetahs, FIP has been described in European wildcats (Felis silvestris), a tiger (Panthera tigris), a mountain lion (Puma concolor), and lion (Panthera leo). This paper reviews the reported cases of FIP in nondomestic felid species and highlights the surveys of FCoV in populations of nondomestic felids. **Intro * Cats are commonly infected with feline coronavirus (FCoV), which can result in the systemic disease feline infectious peritonitis (FIP) * Classified as an alphacoronavirus (species alphacoronavirus-1), FCoV can be grouped into two biotypes: the benign feline enteric coronavirus or the lethal feline infectious peritonitis virus (FIPV). o Also 2 serotypes, type I or type II feline coronavirus * Clinically, cats present with two classic forms of the disease, effusive or noneffusive (‘‘wet’’ or ‘‘dry’’).** Discussion * Credible reports of FIP exist in seven nondomestic felid species, and 15 species have demonstrated seroprevalence. * Antigen shedding, however, is less commonly observed * **Males possibly are at a higher risk compared with females, as observed in domestic cats. ** * The circulation of FCoV in free-ranging species may be ameliorated by the solitary lifestyle of some of these species. * The presence of the virus in the environment is considered relatively short-lived, so transmission is primarily through direct or indirect interactions with either conspecifics or other felids. * In some cases, domestic cats may be a source of prey for larger nondomestic felids. Whether this could pass the virus is unknown but could potentially account for some antibody responses or antigen shedding in the feces * Although antibody testing is useful for detecting previous exposure, even among nondomestic species, a seronegative test does not always predict a lack of viral shedding FIP and FCoV in nondomestic species Cheetah o Serologic response to FCoV first reported in cheetahs in 1979 o Outbreak of FIP at a zoo in Oregon in 1982—nearly 60% of the cheetahs involved in the outbreak succumbed to FIP o Additional cases of FIP have also been described in a litter of three wild-caught, approximately 6-mo-old cheetahs, concurrently infected with Toxoplasma gondii o **Although FIP has undeniably been considered a lethal disease in domestic cats, many hypotheses have been considered regarding what made cheetahs especially susceptible to FIP, including genetic control of the immune response, particularly the major histocompatibility complex homogeneity (MHC)** o Sporadic outbreaks occur, but few devastating losses since the initial outbreak o Discordant results between serology and fecal shedding findings o The role of persistent infections in cheetahs in the development of ulcerative colitis and other long-term sequelae remains less clear and a potential area of needed study European wildcat o In 1993, Watt and colleagues124 reported on an outbreak of FIP in European wildcats in a closed colony that occurred over nearly 14 yr and solely affected males. o Numerous clinical signs were noted, along with multisystemic organ involvement, including the liver, kidneys, intestine, brain, lungs, heart, and lymphoid tissues. o In 2 cats, respiratory signs were noted Mountain lion o A case of FIP has previously been described in a young, male, free-ranging mountain lion in California, with lesions in the small and large intestines, heart, lungs, and brain o Interestingly, the kidneys appeared histologically normal, but viral RNA was evident o This case report is unique in demonstrating FIP in a free ranging animal o Seroprevalence has increased in mountain lions since that time Tiger o FIP has previously been reported in an 8-yr-old, captive male tiger in Romania o Serum amyloid A deposition was also found, which can be due to inflammation but amyloidosis has also been reported in felid species o Seroprevalence studies show ~40% prevalence with no sex or age predilection Lion o First reported in 2 lions in 1970 o more recently, an ocular manifestation of FIP has been described in a 15-yr-old, captive male lion with evidence of blindness o In free-ranging lions in East Africa, 57% of serum samples were considered to demonstrate an antibody response via IFA Serval o Juan-Salle´ s and colleagues54 report two cases of FIP in a group of four servals housed together Lynx species o Several reports in bobcats o Iberian lynx seroprevalence ~25% Leopard o FIP has been reported in leopards in Germany Wild Asian leopard cat o Between 2005 and 2006, 35 out of a total of 1,453 rectal swab samples obtained from Asian leopard cats in wildlife markets were positive for a novel coronavirus, of which the spike protein was found to be phylogenetically similar to that of other alphacoronaviruses, however later evidence suggests it’s closer to the delta coronavirus family Snow leopard o Seroprevalence has been documented but no clinical disease Ocelot o One seropositive wild caught ocelot without clinical disease has been reported Jungle cat o Not confirmed/reported Jaguar o Nott confirmed/reported but FCoV shedding has been documented Caracal o Antibody response has been documented

Answer 78

**Mortality review for the north american snow leopard (Panthera uncia) zoo population from January 1999 to December 2019. ** Womble M, Georoff TA, Helmick K, Carpenter NA, Joslin J, Tupa L, Tetzloff J, McAloose D. Journal of Zoo and Wildlife Medicine. 2021;52(1):145-156 The objective of this 20-yr retrospective study was to review and summarize causes of mortality in the North American (NA) snow leopard population to inform and enhance animal health and husbandry practices. Pathology reports were requested from all NA zoological institutions housing snow leopards that died between 01 January 1999 and 31 December 2019. Data were reviewed and cause of death (COD) and concurrent diseases were summarized and compared by age group, organ system, and disease process. The 241 snow leopards in this report include 109 males, 130 females, and two of undetermined sex. Among them were 116 geriatric snow leopards (>15 yr), 72 adults (15–3 yr), 16 juveniles (3 yr to 2 mo), 32 neonates (2 mo to 0 days), and five fetuses (<0 days). Overall, noninfectious diseases were the most common COD across all age groups (73%). In adult and geriatric snow leopards, chronic renal disease (CRD) (38.8%) and malignant neoplasia (19.7%), including oral squamous cell carcinoma (6.4%), were a common COD. In juveniles and neonates, perinatal death and congenital diseases, including ocular coloboma (15.6%), were a common COD. Individuals with CRD were 13.5 and 4.36 times more likely to have veno-occlusive disease and cardiac fibrosis, respectively. Snow leopards with urolithiasis were 5.27 times more likely to have CRD. Infectious (14.1%) and inflammatory diseases (8.7%) for which no specific etiology was identified were less common overall and more common in juveniles and neonates (25% and 21%, respectively). Neoplasms not previously reported in snow leopards or that are generally uncommon in the veterinary literature included transitional cell carcinoma of the urinary bladder (n = 7) and mesothelioma (n = 1). Background Snow leopard (Panthera uncia) - vulnerable IUCN - Loss of natural prey, habitat fragmentation, climate change, illegal wildlife trade, human-wildlife conflict - Diseases: ocular coloboma, oral and cutaneous papilloma and associated SCC Key Points - 20 yr retrospective ~240 cats, majority geriatric or adult, all captive born, age 0-22.5 yr - Cause of Death, By disease category **Noninfectious most common overall (73%)** - Geriatric and adults: degenerative (DJD esp hips) and neoplastic - Juvenile: anesthesia complication, congenital, endocrine, trauma - Neonates: maternal neglect, rejection, and trauma -- Congenital (30%) -- Multiple Ocular Coloboma most common: eyelid agenesis, PPM, iris colobomas, posterior ocular dysplasia, microphthalmia - **Neoplasia: 58% of geriatric and adult** -- Digestive system (including oral cavity and liver) most common site -- **Oral SCC** most commonly diagnosed malignant tumor ---*Panthera uncia papilloma virus 1 (PuPV-1) confirmed via IHC in one SCC (transformation from cutaneous papilloma) -- Urinary system: transitional cell carcinoma -- Variety of malignant neoplasms not primary COD: oral and sublingual SCC, TCC, thyroid gland adenocarcinoma, endocrine cell carcinoma, cutaneous SCC, renal carcinoma, lymphoma -- Benign tumors: thyroid gland adenoma, meningioma, sublingual papilloma, testicular seminoma, mesothelioma Infectious: bacterial, fungal, viral > parasitic - Bacterial pneumonia and septicemia especially younger - Fungal: systemic, secondary especially older -- Disseminated Scopulariopsis brumptii, phaeohyphomycosis, blastomycosis previously reported - Viral: uncommon, all age groups -- Herpesvirus, feline coronavirus, feline calicivirus, CDV, EEE, FIV - Parasites: enteric, one case of Dirofilaria immitis - single worm contributed to death Inflammatory: - Geriatric: GI (pancreatitis, hepatitis, enteritis) - Adults, juveniles: multisystem - Neonates: respiratory (pneumonia) **Co-morbidities with CRD: veno-occlusive disease, cardiac fibrosis, urolithiasis,** enteritis, parathyroid gland hyperplasia - Urolithiasis: calcium-containing (Ca-Phosphate apatite, Ca carbonate, Ca oxalate) - Veno-occlusive disease: subendothelial fibrosis with occlusion of central or sublobular hepatic veins *monitor liver enzymes along with cardiac and renal function screening - Cardiac fibrosis: suspected from uremic toxins + hypertension **Recommend regular oral exams for papillomas and SCC** Conclusions - Noninfectious disease was more often a cause of death or euthanasia in older animals - Complications of maternal neglect or trauma or infectious disease more common in younger animals - Relationship identified between chronic renal disease and urolithiasis, cardiac fibrosis, and veno-occlusive disease

Answer 79

Journal of Zoo and Wildlife Medicine 52(1): 337–342, 2021 **FIRST REPORT OF ENTEROCYTOZOON BIENEUSI FROM AN AFRICAN LION (PANTHERA LEO) IN A ZOO IN THE REPUBLIC OF KOREA ** Sunwoo Hwang, Jeongho Kim, DVM, PhD, Yu-Jin Park, Dong-Hun Jang, Seung-Uk Shin, Hyung- Chul Cho, and Kyoung-Seong Choi, DVM, PhD ABSTRACT: **Enterocytozoon bieneusi is the most common species of microsporidia that infects humans and animals worldwide**. However, no information is available on E. bieneusi infection among zoo animals in the Republic of Korea (ROK). Here, we investigated the prevalence of E. bieneusi among animals kept in zoos and the zoonotic potential of the E. bieneusi identified. **E. bieneusi was detected only in one African lion (Panthera leo) with diarrhea, using PCR and sequencing analysis of the internal transcribed spacer (ITS) of the rRNA gene. A phylogenetic analysis based on the ITS gene showed that the lion isolate was classified into a novel genotype KPL belonging to Group 2. **The KPL genotype identified in this study differed from genotype I in 6 nucleotides and from genotype I-like in 3 nucleotides, respectively, indicating that Group 2 has the capacity to infect a wide range of hosts. **This is the first report of the presence of E. bieneusi in an African lion housed in a zoo in the ROK.** Further investigation is necessary to study E. bieneusi infection among zoo animals in various regions and to determine the transmission route, in order to control E. bieneusi infection. Study Design: Prospective cross-sectional study (a total of 19 fecal samples from nine wildlife species were screened) Goal: * The objective of this study was to investigate the prevalence of E. bieneusi among zoo-housed animals and to assess the zoonotic potential of E. bieneusi when identified Key Points: * Among the 17 species of microsporidians pathogenic toward humans, Enterocytozoon bieneusi is most commonly found in the GI tract of many animals and causes chronic diarrhea and wasting syndrome o Mainly transmitted through the fecal-oral route via spores from apparently healthy animals * The internal transcribed spacer (ITS) region of the ribosomal RNA gene is used to discriminate the genotyping of E. bieneusi, host specificity, and zoonotic risk o Group 1 has zoonotic potential and poses a major threat to humans, o Groups 2 are mostly host-adapted genotypes associated with specific hosts (ruminants, nonhuman primates, horses, and dogs) and wastewater, suggesting little to no major impact on public health * Of all animals examined, E. bieneusi was detected only in one African lion (Panthera leo) with diarrhea, using PCR of ITS gene o Cryptosporidium parvum and G. duodenalis were not found in this lion * Possible sources of infection include contaminated water and/or food or rodents infected with E. bieneusi * Finding suggests that Group 2 can infect a broad range of hosts and that attention should be focused on the route of transmission and control strategies of E. bieneusi TLDR: * In this study, E. bieneusi infection was detected in an African lion for the first time

Answer 80

Journal of Wildlife Diseases, 57(1): 27-39, 2021 **LEPTOSPIRA PREVALENCE AND ITS ASSOCIATION WITH RENAL PATHOLOGY IN MOUNTAIN LIONS (PUMA CONCOLOR) AND BOBCATS (LYNX RUFUS) IN CALIFORNIA, USA** Mary H. Straub, Jaime L. Rudd, Leslie W. Woods, Deana L. Clifford, and Janet E. Foley ABSTRACT: Leptospirosis is reported infrequently in wild and domestic felids. We estimated the prevalence of Leptospira spp. infection and exposure using real-time PCR and serology, respectively, in 136 mountain lions (Puma concolor) and 39 bobcats (Lynx rufus) that died or were euthanized between 2009 and 2017 from several regions of California, US. Felids were classified as Leptospira-positive if they were test-positive using real-time PCR targeting the LipL32 gene of pathogenic Leptospira spp. or microscopic agglutination test for six serovars of Leptospira spp. **The overall Leptospira spp. prevalence was 46% (63/136) for mountain lions and 28% (11/39) for bobcats.** The most common serovar detected in both felid species was **Leptospira interrogans serovar Pomona**. Age class and geographic location were significantly associated with Leptospira spp. in mountain lions, but not in bobcats. **Interstitial nephritis, predominately lymphocytic, was diagnosed in 39% (41/106) of mountain lions and 16% (4/25) of bobcats evaluated histologically and was significantly associated with being Leptospira spp.-positive in both species.** Our findings suggest that Leptospira spp. infection is common and widespread in California’s wild felids and may have clinical impacts on renal and overall health of individuals. Key Points: - Mountain lion (Puma concolor) and bobcats (Lynx rufus) – population declines due to habitat loss/fragmentation and disease (notoedric mange). Suspect leptospirosis causing morbidity/mortality. -- Opportunistically surveyed sero (MAT) and PCR prevalence of lepto in mountain lion and bobcat (depopulation or injury), and evaluated renal samples for histopathology - Leptospirosis – zoonotic spirochete leading to renal and hepatic dysfunction - Mountain lions: ~50% prevalence of leptospirosis -- Juvenile and mountain lions from southern California had significantly lower seroprevalence -- Interstitial nephritis (primarily lymphocytic) diagnosed in ~40% of individuals --- Increased risk of nephritis if lepto positive (PCR and serology) --- Leptospira interrogans serovar Pomona most common - Bobcats: ~ 30% prevalence of leptospirosis -- Leptospira positive was significantly associated with nephritis on histopath (interstitial nephritis, primarily lymphocytic) --- Leptospira interrogans serovar Pomona most common --- No association with age nor geographic location Take home: - Considerable Leptospira spp PCR and seroprevalence and evidence for Leptospira associated nephritis (interstitial, lymphocytic) among bobcats and mountain lions in California. - Leptospira interrogans serovar Pomona most common. References: None

Answer 81

JZWM 2021 52(3) 918-925 **A RETROSPECTIVE STUDY OF BRAIN LESIONS IN CAPTIVE NONDOMESTIC FELIDS** Abstract: This retrospective study identified and characterized brain lesions in captive nondomestic felids from a large cat sanctuary. Necropsy reports from January 2002 through December 2018 were examined, and gross images and microscopic slides were reviewed from individual cats, where available. In total, 255 cats met the following inclusion criteria: complete necropsy report available, brain examined grossly or microscopically, and age of .1 mon. **Of the 255 cats, 49 cats (19%) were determined to have brain lesions.** Eleven different felid species, as well as one captive-bred hybrid (liger), were included in the study, with tigers (Panthera tigris) (55%) and lions (Panthera leo) (18%) being the most common species. **Lesions were grouped into six etiologic categories: neoplastic (32%), vascular (26%), inflammatory or infectious (20%), congenital (9%), idiopathic (7%), and metabolic (6%).** Not included in these categorized lesions were previously undescribed amphophilic globules in the cerebral cortex of many cats with and without other brain lesions; these were in 95% of lion and 93% of tiger brains where the cerebral cortex was available for histologic examination. These globules were not associated with clinical disease. The histopathologic and gross brain changes documented in this study provide insight into specific diseases and pathologic processes that affect the brains of captive large cat populations. Other than in the abstract: * Male and female evenly distributed * Average age 14.3 years old (1-23 ) * Primary brain neoplasia – meningioma 86% o With or without clinical signs * Ischemic and infarctive lesions = most common vascular lesions * Inflammatory – encephalitis and meningitis – Blastomyces dermatitis most common * Congenital/developmental = chiari-like malformation (lion, tier and bobcat) o Syringomyelia and hydrocephalus– associated with that malformation * Degenerative: gliosis and satelliosis – nonspecific findings, but severe enough to be considered * Metabolic lesions – renal disease -- > uremic encephalopathy * **Undescribed – amphophilic globules present in cerebral cortex of 95% lions, 93% tigers. **Also in liger, caracal, serval and cougars. No visible connection to neurons or glia o **Amphophilic with H&E stain, and positive (blue) with Luxol fast blue stain** Discussion * Lymphoma not identified (2nd CNS neoplasia in cats) * Hypertensive encephalopathy 🡪 neurologic disease o **Unique lesion: meningeal eosinophilic fibrinoid vasculitis with hemorrhage and cerebellar herniation in a 20 yo FI leopard** Take home: amphophilic globules on cerebral cortex not associated with clinical disease.

Answer 82

**BIOMARKERS OF GASTROINTESTINAL DISEASE IN CHEETAHS (ACINONYX JUBATUS). ** Fox L, Haefele H, Uelmen J, Hoppes S, Swenson J, Tolbert MK, Suchodolski JS, Steiner JM. Journal of Zoo and Wildlife Medicine. 2021;52(3):886-892. Gastrointestinal disease is a common clinical problem in captive cheetahs (Acinonyx jubatus). It is reported that gastritis affects the vast majority of the captive population of cheetahs. Pancreatitis and acute and chronic enteritis have also been reported. These issues pose significant long-term health and welfare implications for cheetahs. Cobalamin, folate, methylmalonic acid (MMA), gastrin, feline pancreatic-specific lipase immunoreactivity (fPLI), and feline trypsin-like immunoreactivity (fTLI) immunoassays are important biomarkers of gastrointestinal disease in domestic cats. **The goal of this study was to determine if these immunoassays validated in domestic cats could be used clinically in cheetahs, by establishing reference intervals (RI) for these biomarkers in cheetahs. A cohort of 40 clinically healthy cheetahs was selected from three zoological institutions on the basis of being free of clinical gastrointestinal disease and extra-gastrointestinal disease that could affect biomarkers, as well as having banked frozen serum.** Cheetah biomarker RI, with domestic cat RI for comparison in parentheses, are as follows: cobalamin 470–618 pg/ml (290–1500 pg/ml), folate 2.2–15.7 ng/ml (9.7–21.6 ng/ml), MMA 365–450 nM/L (139–897 nM/L), fPLI 0.5–1.2 µg/L (0–4 µg/L), and gastrin 30–50 pg/ml (<10–39.5 pg/ml). This study shows that RI for gastrointestinal biomarkers can be notably different, even between species that are as closely related as the domestic cat and the cheetah. Additionally, it was found that the fTLI assay does not cross-immunoreact with cheetahs. In conclusion, this study emphasizes the importance of developing species-specific RI for biomarker assays and using caution when extrapolating RI from other species. Background Cheetahs - IUCN ‘vulnerable’, M&M predominantly chronic degenerative disease (not infx) - Glomerulosclerosis, veno-occlusive disease, chronic lymphoplasmacytic gastritis, systemic amyloidosis - Higher cortisol concentrations and larger adrenal cortices compared to wild cheetahs suggests chronic stress as a factor Reported GI diseases: pancreatitis, enteritis, astrovirus, coronavirus, Clostridium - 80% of captive cheetahs have histologic changes of gastritis - Helicobacter spp. suspected to contribute - wild cheetahs have similar infx but only 3% gastritis **Folate (vitamin B9):** water-soluble, co-factor for amino acid and vitamin synthesis, role in DNA replication, repair, and methylation, deficiency in cats causes poor appetite, growth, and bone marrow changes - **Reduced with proximal small intestinal disease** - Reduced storage with hepatic disease and reduced excretion with renal disease - Increased with small intestinal dysbiosis (bacteria synthesize folate) **Cobalamin (vitamin B12):** water-soluble, synthesized by microbiota in herbivores, carnivores depend on dietary intake - Role in DNA synthesis - deficiency affects rapidly dividing cells (GI, hematopoietic), can cause GI disease, central and peripheral neuropathies, immunodeficiencies, and myelopathies - **Reduced with ileum disease (distal SI) from destruction of ileal cobalamin receptors** - Reduced in EPI from carrier protein deficiency (intrinsic factor) and pancreatic protease deficiency that would normally digest cobalamin-binding protein (R protein) - Reduced with small intestinal dysbiosis Methylmalonic Acid (MMA) - more formation when cells are deficient in cobalamin - marker of cellular cobalamin deficiency - (Cobalamin-dependent reactions occur intracellularly so MMA is a better indicator than serum cobalamin) - Excreted through kidneys so interpret cautiously with kidney disease Gastrin - peptide hormone produced by G cells in stomach and proximal duodenum, stimulates gastric acid production, has trophic efects on gastric mucosa - Mildly increased with gastritis or IBD in dogs and CKD in cats - Marked increases suggest gastrinomas - Increased after PPI administration fPLI - measures pancreatic lipase - Elevation indicates acinar cell damage most likely from pancreatitis - Chronic pancreatitis can lead to acute panc, DM, or EPI - False negatives possible in cats with mild or chronic panc from fibrosis and atrophy that doesn’t cause leakage of pancreatic enzymes TLI - trypsin and trypsinogen from pancreatic acinar cells - Severely decreased with EPI (CS: impaired nutrient absorption, soft, voluminous stools, weight loss, vitamin deficiencies) Key Points - Retrospective from 3 zoos, some had gastric biopsies opportunistically - cheetahs with mod-severe or worse gastritis were excluded - No correlation of any biomarker with sex - Decreasing fPLI correlated with increasing age but low fPLI is not clinically relevant - No statistical correlation between cobalamin and MMA - Couldn’t establish RI for fTLI because all were lower than assay range - lack of species cross-immunoreactivity for this assay - 6 cheetahs with mod-severe gastritis had high serum gastrin concentrations Conclusions - fTLI assay does not cross-immunoreact with cheetahs - MMA did not correlate with cobalamin - Decreasing fPLI with increasing age in cheetahs - Cheetahs with gastritis may have high serum gastrin - RI for GI panels established in cheetahs

Answer 83

Mathieu, A., & Garner, M. M. (2021). **A RETROSPECTIVE STUDY OF NEOPLASIA IN NONDOMESTIC FELIDS IN HUMAN CARE, WITH A COMPARATIVE LITERATURE REVIEW. ** Journal of Zoo and Wildlife Medicine, 52(2), 413-426. Abstract: This retrospective study of neoplasia in nondomestic felids in human care presents the cases diagnosed at Northwest ZooPath (NWZP), Monroe, Washington, from 1998 to 2017 in conjunction with a scoping literature review. The 554 neoplasms identified in 20 species in the NWZP archive were combined with the 984 neoplasms identified in those same species in the published literature. Some of the cases identified in the literature were from the NWZP archive. **Based on this review, mammary adenocarcinoma (183/1,483, 12.3%), lymphoma (89/1,483, 6.0%), squamous cell carcinoma (85/1,483, 5.7%), pheochromocytoma (57/1,483, 3.8%), and thyroid adenoma (57/1,483, 3.8%) are the most frequently reported neoplasms in nondomestic felids in human care.** *Apparent species predilections for neoplasia include mammary adenocarcinoma in tigers, jaguars, lions, and jungle cats; lymphoma in lions and tigers; squamous cell carcinoma in snow leopards; pheochromocytoma in clouded leopards; ovarian adenocarcinoma in jaguars; cholangiocarcinoma in lions and tigers; multiple myeloma in tigers; bronchoalveolar adenocarcinoma in cougars and lions; hemangiosarcoma, hepatocellular carcinoma, and gastrointestinal adenocarcinoma in lions; mesothelioma in clouded leopards, lions, and tigers; myelolipoma and cutaneous mast cell tumor in cheetahs; soft tissue sarcomas in tigers; and transitional cell carcinoma of the urinary bladder in fishing cats.* Key Points: **Mammary adenocarcinoma, lymphoma, SCC, pheochromocytoma, thyroid adenoma most frequently reported neoplasms in nondomestic felids in human care.** Not previously reported in the literature: - African wildcat mammary adenocarcinoma. - Arabian sand cat parathyroid carcinoma and thyroid carcinoma. - Black-footed cat lacrimal gland adenocarcinoma. - Pallas’s cat – Hemangiosarcoma, TCC of urinary bladder. - Serval – Mammary adenocarcinoma, cutaneous/subcutaneous lipomas most common. By Species - Bobcats – Endocrine/neuroendocrine neoplasms most commonly reported. Lymphoma and thyroid adenoma most commonly reported neoplasias. - Cheetahs – Splenic myelolipoma most common followed by hepatic myelolipoma, MCT, and reproductive leiomyoma. - Clouded leopards – Pheochromocytoma most common, followed by mesothelioma, lymphoma, repro leiomyoma. - Cougar (puma, mtn lion) – Thyroid adenoma most common followed by thyroid carcinoma, mammary adenocarcinoma, cholangioadenoma, lymphoma, bronchoalveolar adenocarcinoma. - Fishing cat – Transitional cell carcinoma of the urinary bladder most common. - Jaguar – Ovarian adenocarcinoma most common followed by mammary adenocarcinoma, uncharacterized repro neoplasia, repro leiomyoma. - Jungle cat – Mammary adenocarcinoma most common. - Leopard – Reproductive leiomyoma is the most common followed by thyroid carcinoma, cholangiocarcinoma, parathyroid adenoma, mammary adenocarcinoma, and cutaneous/subcutaneous lipoma. - Leopard cat (Prionailurus bengalensis) – Lymphoma most common. - Lions – Lymphoma most common, followed by mammary adenocarcinoma, repro leiomyoma, mesothelioma, bronchoalveolar adenocarcinoma, soft tissue sarcoma. - Lynx – SCC most common (integumentary most common but also had GI and resp forms). - Ocelot – Oral SCC, TCC of urinary bladder, and metastatic carcinoma of undetermined lineage most common. - Snow leopard – Oral and cutaneous papillomas most common followed by SCC, thyroid adenoma, seminoma. Digestive neoplasms most common overall. - Tiger – Mammary adenocarcinoma followed by repro leiomyomas, soft tissue sarcomas, thyroid adenomas, lymphoma, SCC. **Mammary neoplasms are the most frequently reported neoplasms in nondomestic felids in human care.** - Tigers, jaguars, lions, jungle cats. - Malignancy rate in nondomestic felids is high (97%). - Prolonged exposure to progestin contraceptive (MGA) linked to mammary adenocarcinomas in nondomestic felids, with resolution of hyperplastic lesions after contraceptive was removed. - AZA recommends use of GnRH agonists, early ovariohysterectomy, or ovariectomy and warns against the long term use of MGA. **Lymphoma second most common.** - Bobcats, lions, tigers. - CS – Intermittent inappetence to anorexia, lethargy, wt loss, splenomegaly, lymphadenopathy. - Often multicentric in lions and tigers, most commonly involving the spleen, liver, LN. - Lymphomas in lions never involved the GIT. o Prognosis is grave, but remission can be achieved. **SCC (GI or integumentary) third most common.** - Snow leopards, lynx, tigers. - Cutaneous papillomas reported here for the first time in cheetahs. **Pheochromocytoma and thyroid adenoma equal for 4th most common**. - Pheo – Clouded leopards, was often an incidental finding. - Functional pheos can result in systemic hypertension, malignant variants can met or locally invade the vena cava. - It has been proposed that hypertension assoc with pheos could lead to diffuse follicular telogenization and development of the clouded leopard alopecia syndrome. Thyroid adenomas most common in tigers, usually incidental. Other species predilections: o Jaguar ovarian adenocarcinoma. o Lion and tiger cholangiocarcinoma. o Tiger multiple myeloma. o Cougar and lion bronchoalveolar adenocarcinoma. o Lion hemangiosarc, hepatocellular carcinoma and GI adenocarcinoma. o Clouded leopard, lion, and tiger mesothelioma. o Cheetah myelolipoma and cutaneous MCT. o Tiger soft tissue sarcomas. o Fishing cat TCC. Most commonly reported neoplasms in Felinae – Hemolymphatic, endocrine/neuroendocrine, and integumentary. Most commonly reported neoplasms in Pantherinae – Repro, endocrine/neuroendocrine, mammary.

Answer 84

Journal of Wildlife Diseases, 57(4), 2021, pp. 784–798 **PSEUDORABIES (AUJESZKY’S DISEASE) IS AN UNDERDIAGNOSED CAUSE OF DEATH IN THE FLORIDA PANTHER (PUMA CONCOLOR CORYI)** ABSTRACT: Feral swine (Sus scrofa), an important prey species for the endangered Florida panther (Puma concolor coryi), is the natural host for pseudorabies virus (PRV). Prior to this study, PRV had been detected in just three panthers. **To determine the effect of PRV on the panther population, we prospectively necropsied 199 panthers and retrospectively reviewed necropsy and laboratory findings, reexamined histology, and tested archived tissues using real-time PCR from 46 undiagnosed panther mortalities. **Seven additional infections (two prospective, five retrospective) were detected for a total of 10 confirmed panther mortalities due to PRV. To further evaluate the effect of PRV, we categorized radio-collared (n = 168) and uncollared panther mortalities (n = 367) sampled from 1981 to 2018 based on the likelihood of PRV infection as confirmed, probable, suspected, possible, or unlikely/negative. Of 168 radio-collared panthers necropsied, PRV was the cause of death for between eight (confirmed; 4.8%) and 32 (combined confirmed, probable, suspected, and possible categories; 19.0%) panthers. **The number of radio-collared panther mortalities due to PRV was estimated to be 15 (95% empirical limits: 12–19), representing 8.9% (confidence interval: 4.6–13.2%) of mortalities. Gross necropsy findings in 10 confirmed cases were nonspecific. Microscopic changes included slight to mild perivascular cuffing and gliosis (primarily in the brain stem), lymphoplasmacytic meningoencephalitis (cerebral cortex), and intranuclear inclusion bodies (adrenal medulla).** The PRV glycoprotein C gene sequences from three positive panthers grouped with the sequence from a Florida feral swine. Our findings indicate that PRV may be an important and underdiagnosed cause of death in Florida panthers Study Design: Retrospective and prospective cross sectional epidemiological study Goal: Describe the epizootiology of PRV in the panther population and determine the importance of PRV infection as a cause of death in Florida panthers Key Points: - Swine are the definitive hosts of suid alphaherpesvirus 1 (pseudorabies virus, PRV), which causes pseudorabies (Aujeszky’s disease) in swine and other mammals -- Infections with PRV in carnivores, ruminants, and rodents are typically fatal -- The PRV Eradication Program eliminated PRV from US domestic swine in 2004 - The seroprevalence of PRV in feral swine in Florida varies but can reach 79% -- In feral swine, most PRV infections are latent/subclinical -- Stressors such as concomitant disease, farrowing, and breeding activity can reactivate infection, leading to viral shedding and subsequent risk of transmission -- Transmission to panthers probably occurs by ingestion and is supported by the apparent point source cause of death in what appeared to be a family group of three panthers - An estimated 9% of radio-collared panther mortalities were due to PRV -- Third leading diagnosed cause of death, behind intraspecific aggression and vehicular collision -- Telemetry data, gross, and microscopic pathology indicated acute death (<48h) - Infection with PRV in felids is often focal and unilateral and the diagnosis may be missed - Low sensitivity in positive controls underscores the difficulty in diagnosing PRV -- A negative real-time PCR alone does not necessarily rule out infection - Confirmed cases were especially concentrated in and around game enclosures that contained high densities of feral swine - Management to increase white-tailed deer (Odocoileus virginianus) density could decrease the risk of PRV to panthers -- Additionally, feral swine management such as vaccination, harvest management targeting higher risk older swine, reduction of swine densities, and restriction of the use of dogs for hunting feral swine in panther range may decrease the prevalence of PRV or of viral shedding -- These management actions would be most practical, and likely more effective, in closed feral swine populations TLDR: - Feral swine = important prey species for the endangered Florida panther and natural host for suid alphaherpesvirus 1 (pseudorabies virus, PRV) - PRV causes pseudorabies (Aujeszky’s disease) which is fatal in carnivores, ruminants, and rodents - PRV may be an important and underdiagnosed cause of death in Florida panthers

Answer 85

Anis, E., Needle, D. B., Stevens, B., Yan, L., & Wilkes, R. P. (2020). **Genetic characteristics of canine distemper viruses circulating in wildlife in the united states. ** Journal of Zoo and Wildlife Medicine, 50(4), 790-797. Abstract: Canine distemper virus (CDV) is a highly contagious disease of wild and domestic mammals. Maintenance of CDV among wildlife plays an important role in the disease epidemiology. **Wild animals, including raccoons (Procyon lotor) and gray foxes (Urocyon cinereoargenteus), serve as reservoirs of CDV** and hamper the control of the disease. Recently, we discovered that at least **three different CDV lineages (America-3 [Edomex], America-4, and America-5] that are genetically different from the available vaccine strains are circulating in domestic dogs in the United States**. Because wildlife serve as a reservoir for the virus, it is important to determine if wildlife play a role in the maintenance and spread of these lineages. To determine the genetic characteristics of circulating strains of CDV in wildlife in various geographic regions in the United States, we studied the nucleotide sequences of the hemagglutinin (H) gene of 25 CDV strains detected in nondomestic species. The species included were free-ranging wildlife: three fishers (Martes pennanti), six foxes, one skunk (Mephitis mephitis), 10 raccoons, two wolves (Canis lupus), and one mink (Neovison vison). Strains from two species in managed care, one sloth (Choloepus didactylus) and one red panda (Ailurus fulgens), were also evaluated. **Phylogenetic analysis of the H genes indicated that in addition to America-3, America-4, and America-5 lineages, there are at least two other lineages circulating in US wildlife.** One of these includes CDV nucleotide sequences that grouped with that of a single CDV isolate previously detected in a raccoon from Rhode Island in 2012. The other lineage is independent and genetically distinct from other CDV strains included in the analysis. **Additional genetically variable strains were detected, mainly in raccoons, suggesting that this species may be the host responsible for the genetic variability of newly detected strains in the domestic dog population.** Background: - CDV – Morbillivirus, Paramyxoviriae, ssRNA virus. - Detected in all families of terrestrial carnivores – Canidae, Felidae, Mustelidae, Ursidae, Vivveridae, Hyaenidae, Procyonidae. Multihost nature, unlike some other morbilliviruses i.e. measles. - Respiratory, CNS, GI signs. - High mortality in juveniles. - Transmission via direct contact, air, bodily secretions, +/- fomites. - Mutations affecting the viral signaling lymphocytic activation molecule SLAM receptor binding region affects the ability of the virus to infect new species. - Maintenance within wildlife reservoirs between outbreaks hampers disease eradication and plays important role in epidemiology in domestic dogs. - Six CDV sequences that were detected in fox, raccoon, red panda in SE grouped with America-5 lineage. Reported in domestic dogs. -- The red panda was a zoo animal, three other RPs died of distemper 1 month prior. -- Was vaccinated once with RECMBITEK vaccine, need multiple boosters for protection. -- Captive sloth from TN where another outbreak took place with American 4 lineage. -- Three nucleotide sequences grouped with American-3 (Edomex) lineage. -- Multiple CDV strains simultaneously circulating in domestic dogs and multiple wildlife species, adds to complexity of understanding epidemiology of CDV. -- Likely that raccoons are the source of ‘new’ genetic lineages. Takeaway: Recent rise in CDV cases in wildlife and detection of the same lineages circulating in domestic dogs suggests currently available vaccines are insufficient to keep these strains confined to reservoir populations. CDV distribution and epidemiology is complex.

Answer 86

**Glomerular filtration rate determined by measuring serum clearance of a single dose of inulin and serum symmetric dimethylarginine concentration in clinically normal cheetahs (Acinonyx jubatus).** Sanchez, C.R., Hayek, L.A.C., Carlin, E.P., Brown, S.A., Citino, S., Marker, L., Jones, K.L. and Murray, S. American journal of veterinary research, 2020;81(4):375-380. OBJECTIVE To establish a reference interval for glomerular filtration rate (GFR) determined by measuring serum clearance of a single IV dose of inulin in clinically normal cheetahs (Acinonyx jubatus) and compare serum symmetric dimethylarginine (SDMA) concentration in cheetahs with GFR. ANIMALS 33 cheetahs housed at 3 institutions. PROCEDURES A single bolus of inulin (3,000 mg/m2) was administered IV, and 5 serial blood samples were collected and analyzed for serum inulin concentration with the anthrone technique. The GFR was estimated with a modified slope-intercept method for the slow component of the serum concentration-versus-time curve. Blood urea nitrogen and serum creatinine concentrations were measured in samples obtained immediately prior to inulin administration, and serum SDMA concentration was measured in stored samples. RESULTS Mean ± SD measured GFR was 1.58 ± 0.39 mL/min/kg, and the calculated reference interval was 0.84 to 2.37 mL/min/kg. **There were significant negative correlations between GFR and serum creatinine concentration (r = −0.499), BUN concentration (r = −0.592), and age (r = −0.463). Serum SDMA concentration was not significantly correlated with GFR (r = 0.385), BUN concentration (r = −0.281), or serum creatinine concentration (r = 0.165).** CONCLUSIONS AND CLINICAL RELEVANCE A reference interval for GFR in clinically normal cheetahs was obtained. Further evaluation of animals with renal disease is needed to determine whether measuring serum clearance of a single IV dose of inulin is a reliable diagnostic test for early detection of renal disease in cheetahs. Background - Cheetah - vulnerable IUCN, CKD: one of leading causes of death in captive populations - Glomerulosclerosis and nephrosclerosis in up to 90% of study populations - AKI and CKD cause of death in 25% of 1 study pop - Postulated risk factors: reduced genetic diversity, diet, age, chronic stress, renal medullary fibrosis - SDMA: >90% elimination by glomerular filtration, earlier elevations with CKD in 1 cheetah study - Reference interval has not been established Key Points - Measured GFR with exogenous inulin (IV under GA): RR 0.84-2.37 mL/min/kg - No adverse effects reported - Significant negative correlation between GFR, serum crea, serum BUN, and age - Serum crea higher in females than males - 24 stored serum samples: SDMA was not significantly correlated with GFR, crea, BUN, or increase with age - Past studies: 92 stored samples, high correlation between SDMA and serum crea in cheetahs with renal disease Conclusions - GFR measurement via inulin injection, reference range established for cheetahs. - GFR by inulin had high correlation with BUN, creatinine, and age. - Despite past studies, serum SDMA in this study did not significantly correlate with GFR, crea, BUN, or age.

Answer 87

**EVALUATION OF TWO DOSES OF BUTORPHANOL- MEDETOMIDINE-MIDAZOLAM FOR THE IMMOBILIZATION OF WILD VERSUS CAPTIVE BLACK- FOOTED CATS (FELIS NIGRIPES).** JZWM 2020 51(3) 497-506. Eggers, Birgit, Tordiffe, Adrian S. W., Lamberski, Nadine, Lawrenz, Arne, Sliwa, Alexander, et al. The efficacy, safety, physiologic effects, and reversibility of butorphanol-medetomidine-midazolam (BMM) immobilization were evaluated in black-footed cats (Felis nigripes) and compared between captive and wild animals. Nine captive and 14 wild black-footed cats were hand injected into an accessible hind limb muscle group with the BMM combination. **The captive cats (captive group) received a lower dose of the combination (butorphanol, 0.25 6+/- 0.03 mg/kg; medetomidine, 0.06 +/- 0.01 mg/kg; midazolam, 0.13 +/- 0.02 mg/kg), whereas the wild cats received a higher dose (butorphanol, 0.53 +/- 0.11 mg/kg, medetomidine, 0.13 +/- 0.03 mg/kg, midazolam, 0.27 +/- 0.05 mg/kg).** Two capture methods were required to restrain the wild cats; previously collared cats were tracked and excavated out of their burrows during daylight hours (excavated group), whereas uncollared cats were randomly located using spotlights and pursued by a vehicle at night (pursued group). Inductions were rapid and no spontaneous arousals occurred. **Mean arterial blood pressure in all cats was within normal limits for domestic cats**. Initial rectal temperatures varied greatly among the groups, but decreased in all groups as the immobilization progressed. In the pursued animals, heart rates and respiratory rates were initially elevated. **All cats had moderate hypoxemia, hypocapnia, and metabolic acidosis**. Intramuscular administration of naltrexone, atipamezole, and flumazenil resulted in rapid, uncomplicated recoveries. BMM is thus a safe, effective immobilizing drug combination for both captive and wild black-footed cats, but higher doses are required in wild animals. The capture methods exerted a greater influence on the physiology of the immobilized animals than did the doses of the drugs used. Although this drug combination can be used safely to immobilize black-footed cats, supplemental oxygen should always be available for use, especially in pursued animals due to hypoxia. Background, study design - Smallest wild felid species of the southern African, predisposed to renal amyloidosis, no published immobilization protocols - N = 9 captive, 14 wild: wild group immobilized by either excavation from burrows or pursued - Captive BFC received 0.2mg/kg butorphanol + 0.05mg/kg medetomidine + 0.1mg/kg midazolam, wild BFC doses were doubled. Reversed with atipamezole, flumazenil, naltrexone Key points - **Rapid induction with no sudden arousals (occurs in cheetahs), rapid uncomplicated recovery with reversals, longer immobilizations or more invasive may require higher doses** - Higher flumazenil dose required for wild cats (higher required from pilot study for all groups) - No vomition occurred (common effect of medetomidine in domestic cats), possibly aided by reduced medetomidine effects from butorphanol (domestic cats) or antiemetic effects of midazolam (people) - **Blood pressure within normal limits for domestic cats, no difference between groups. No hypotension important as BFC predisposed to renal amyloidosis**. - HR higher in pursued cats over entire immobilization than captive cats - RR higher than normal range for domestic cats and anesthetized servals - ALL had hypoxemia, hypocapnia, metabolic acidosis, worse in pursued cats from hyperelactemia, likely from increased O2 utilization with exertion and peripheral vasoconstriction (medetomidine) - Hyperlactemia correlated with distance pursued - ALL hyperglycemic but not pathologic, secondary to fight-or-flight and ↓insulin release by medetomidine Take home: BMM rapid induction and recovery with wild (double doses) and captive BFC. All hypoxemic, hypocapnic, metabolic acidodic, worse in pursued cats

Answer 88

**SARS-CoV-2 Infection And Longitudinal Fecal Screening In Malayan Tigers (Panthera tigris jacksoni), Amur Tigers (Panthera tigris altaica), And African Lions (Panthera leo krugeri) At The Bronx Zoo, New York, USA. ** Bartlett, S.L., Diel, D.G., Wang, L., Zec, S., Laverack, M., Martins, M., Caserta, L.C., Killian, M.L., Terio, K., Olmstead, C. and Delaney, M.A. Journal of Zoo and Wildlife Medicine, 2020;51(4):733-744. Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) emerged as the cause of a global pandemic in 2019–2020. In March 2020, New York City became the epicenter in the United States for the pandemic. On 27 March 2020, a Malayan tiger (Panthera tigris jacksoni) at the Bronx Zoo in New York City developed a cough and wheezing with subsequent inappetence. Over the next week, an additional Malayan tiger and two Amur tigers (Panthera tigris altaica) in the same building and three lions (Panthera leo krugeri) in a separate building also became ill. The index case was anesthetized for diagnostic workup. Physical examination and bloodwork results were unremarkable. Thoracic radiography and ultrasonography revealed a bronchial pattern with peribronchial cuffing and mild lung consolidation with alveolar-interstitial syndrome, respectively. SARS-CoV-2 RNA was identified by real-time, reverse transcriptase PCR (rRT-PCR) on oropharyngeal and nasal swabs and tracheal wash fluid. Cytologic examination of tracheal wash fluid revealed necrosis, and viral RNA was detected in necrotic cells by in situ hybridization, confirming virus-associated tissue damage. SARS-CoV-2 was isolated from the tracheal wash fluid of the index case, as well as the feces from one Amur tiger and one lion. Fecal viral RNA shedding was confirmed in all seven clinical cases and an asymptomatic Amur tiger. Respiratory signs abated within 1–5 days for most animals, although they persisted intermittently for 16 days in the index case. Fecal RNA shedding persisted for as long as 35 days beyond cessation of respiratory signs. This case series describes the clinical presentation, diagnostic evaluation, and management of tigers and lions infected with SARS-CoV-2 and describes the duration of viral RNA fecal shedding in these cases. This report documents the first known natural transmission of SARS-CoV-2 from humans to nondomestic felids. Background - SARS-CoV-2 - emergence from live wild animal market, 96% homology with BatCoV RaTG13 - SARS-CoV-like viruses isolated from or detected in Himalayan palm civets and raccoon dogs - SARS-CoV-1 infection demonstrated in Chinese ferret badgers, ferrets, cynomolgus macaques, rhesus macaques, and African green monkeys - SARS-CoV-2 shedding in 2 naturally exposed dogs in Hong Kong - Natural infection with respiratory signs in farmed mink in Netherlands and Denmark - Only other confirmed infection in nondomestic felid: puma at a zoo in South Africa Key Points - Dry-to-wet cough, wheezing, decreased appetite, possibly vomiting, epistaxis - Short course of disease: complete resolution in 1-16 days - Rads: Generalized bronchial pattern with multifocal caudal peribronchial cuffing - TFAST: B-lines (consolidated peripheral lung and coalescing B-lines) - Tracheal wash: tracheal epithelial necrosis - RNA detected by rt-PCR of tracheal wash, oropharyngeal swab, and nasal swab, virus isolation from tracheal wash - Immunofluorescent staining for antigen, in situ hybridization - Seroconversion demonstrated on virus neutralization assay * Fecal rRT-PCR: shedding duration varied greatly by individual (5-30+ days past end of CS) - Virus isolation on feces (confirms viable virus present - source of infection) - Experimentally domestic cats stop shedding within 1 week - 1 Lion stopped shedding for 2 weeks then started again - Tigers and lions were infected by different viral genotypes suggesting unrelated transmission events (different keeper exposures) - Transmission likely occurred during times keepers were asymptomatically shedding virus - Transmission from fomites/aerosol possible: tigers housed individually but rotated through shared enclosures - No long term complications noted yet Conclusions - Confirmed susceptibility of tigers and lions to SARS-CoV-2 - Initial infection route for tigers and lions appears to be from different keepers who were asymptomatically shedding - Clinical course generally short (5 days), fecal shedding up to 35 days after end of respiratory signs - Fecal viral RNA shedding suggests viral replication in the GI tract - 1 asymptomatic shedding tiger - PPE should be used to minimize anthropozoonotic transmission of coronavirus to Felidae and other susceptible taxa - SARS-CoV-2 is an OIE reportable disease, testing should be coordinated with state regulatory agencies

Answer 89

**CONTRAST-ENHANCED COMPUTED TOMOGRAPHIC FINDINGS OF APPARENTLY CLINICALLY NORMAL CHEETAH (ACINONYX JUBATUS) LIVERS** Jania and Kinney, DVM, DACZM et al. - Journal of Zoo and Wildlife Medicine 52(1): 320–326, 2021 Abstract: Hepatic veno-occlusive disease (VOD) is a significant cause of morbidity and mortality in captive cheetahs (Acinonyx jubatus), and the appearance of this disease in humans by computed tomography (CT) has been well described. **Contrast-enhanced CT abdominal scans of cheetahs without evidence of hepatic disease (n = 5) were reviewed retrospectively to describe the normal appearance of cheetah livers as an aid to antemortem VOD diagnosis.** Despite having no clinical signs, clinical pathology abnormalities, or hepatic biopsy histopathology supportive of VOD, **all five cheetahs had at least one VOD consistent finding on CT**. The results of this study suggest that given the progressive and potentially subclinical nature of VOD, CT could serve as a noninvasive screening tool and be used to monitor disease progression. Background: Captive cheetahs prone to hepatic veno-occlusive disease – up to 83% prevalence in North America and primary cause of death in 30% of animals in one study, and incidental finding in others - VOD – microvascular deposition of fibrous material impairing normal flow of hepatic sinusoids - Causes hepatic venous outflow obstruction 🡪 intrahepatic portal hypertension and hepatic failure - Definitive etiology unknown, but may be caused by diet, toxins, infectious disease, and stress - Analogous to SOS (sinusoidal obstruction syndrome in people) - Biopsy gold standard for most hepatic diseases - CT used as noninvasive evaluation of liver and can help to r/o disease - Unlike US – CT is not user dependent, the entire liver can be imaged in large animals, and dynamic scans can be performed for information on vascular and perfusion patterns Details: CT of 5 cheetahs revealed degree of contrast enhancement, gallbladder wall thickness (1.4 mm), biliary tree appearance, hepatic and portal vasculature branching patterns consistent with canine publications - Several had findings consistent with human VOD: -- 3 had varying degrees of heterogenous contrast enhancement with ill-defined, irregularly shaped areas of poor enhancement (not normal in dogs) -- All 5 had varying degrees of periportal hypoattenuation vasculature -- Not described in vet med – humans associated with hepatic congestion or periportal inflammation - One had “nutmeg” liver appearance - R lateral lobe extended further caudally in all animals and one had majority of liver positioned to the right - 4 main tributaries made up extrahepatic portal vein: cranial and caudal mesenteric veins, the splenic vein, and gastroduodenal veins - R and L branch of portal v.: R supplies R lateral & caudate lobes, left supplying R medial, quadrate and L lobes - Caudal vena cava had 2 main branches: R supplying quadrate and R liver lobes and L supplying L hepatic lobes - VOD can be focal or diffuse and false-negative biopsies are possible Take-away: - All 5 cheetahs in this study had CT findings consistent in people with VOD despite lack of clinical signs, clin path abnormalities, or consistent hepatic biopsies. - CT, if validated, could be a noninvasive screening tool for subclinical or suspected cases of VOD.

Answer 90

JZWM 2021 52(3) 1079-1083 **EXOCRINE PANCREATIC INSUFFICIENCY-LIKE SYNDROME IN FOUR CAPTIVE TIGERS (PANTHERA TIGRIS)** Abstract: **Exocrine pancreatic insufficiency (EPI) is a condition characterized by a decreased synthesis and secretion of pancreatic enzymes, which results in weight loss, poor hair coat, and diarrhea. The diagnostic test of choice for EPI in domestic cats is feline serum trypsin-like immunoreactivity (fTLI).** This paper details four tigers (Panthera tigris) with clinical signs compatible with EPI. On the basis of domestic cat reference ranges, fTLI assays for all four clinically affected tigers were diagnostic for EPI (median 1.0 lg/L; range 0.5–1.2 lg/L). All four tigers had a rapid clinical response to pancreatic enzyme supplementation. Serum from 10 clinically healthy tigers was submitted for the fTLI assay, for comparative purposes. The **healthy tigers’ fTLI assays were also within range for a diagnosis of EPI in domestic cats (median 3.1 lg/L; range 1.9–4.5 lg/L); however, clinically affected tigers had significantly lower serum fTLI concentrations than healthy tigers (P 1⁄4 0.0058). Serum cobalamin was below the detection limit in both the affected and healthy tigers (,150 ng/L).** Measuring fTLI appears to be a useful tool in the diagnosis of EPI-like syndrome in tigers. As in other species, EPI-like syndrome in tigers may also be associated with cobalamin deficiency. Key Points: - EPI = sequela to chronic pancreatitis -- > decreased secretion of pancreatic enzymes - Steatorrhea, undigested food, increased appetite, poor haircoat - Antemortem- pancreatic function testing: fTLI - Tx: exogenous enzyme supplementation and cobalamin if needed Case series: 1 female 3 males with unformed, malodorous stools - CBC normal in 2 animals, 2 had leukocytosis and neutrophilia - Rads unremarkable, U/S – 2 animals with increased echogenicity of S.I muscularis - Endoscopy – no gross abnormalities – biopsies = lymphoplasmacytic gastritis - fTLI, cobalamin and folated tested, compared to 10 clinically healthy tigers - Affected – significantly lower fTLI, and folate were significantly hiher. Cobalamin below detection levels (also for healthy animals) - Pancreatic enzymes started – imporved coat condition and stool quality within a week - One animal euthanized with vertebral osteomyelitis (unrelated) - Histo: pancreatic lipomatosis with slight chronic pancreatitis Discussion - fTLI highly specific for pancreatic function - EPI – developed until >90% loss of acinar function - fTLI – measures trypsinogen and trypsin which is increase during pancreatic inflammation - Hypocobalaminemia is common in domestic cats -- Absorption is dependent to intrinsic factor produced by pancreas -- Can cause primary GI inflammation - Serum folate were significantly greater in affected tigers -- Due to intestinal dysbiosis and increase folate synthesis, increased folate absorption form lowered duodenal pH or decreased activation to a useable form because of hypocobalaminemmia Take home: fTLI and pancreatic supplementation can be done in tigers.

Answer 91

**CYTOKINE-RELEASE ASSAY FOR THE DETECTION OF MYCOBACTERIUM BOVIS INFECTION IN CHEETAH (ACINONYX JUBATUS)** JZWM 52(4), 2021 Abstract: The lack of species-specific assays for the diagnosis of infectious diseases, such as bovine tuberculosis, poses a threat to the management of wildlife populations, especially for vulnerable species such as cheetah (Acinonyx jubatus). **The aim of this study was to identify and develop a cell-mediated immunological cytokine-release assay that could distinguish between Mycobacterium bovis–infected and uninfected cheetahs using commercially available feline cytokine ELISA and domestic cat (Felis catus) recombinant proteins**. Antibodies against domestic cat cytokines, tumour necrosis factor alpha (TNF-a), interleukin-1 beta (IL-1b), and interferon gamma (IFN-c), were screened for cross-reactivity with plasma cytokines from cheetah whole blood stimulated using QuantiFERONt-TB Gold Plus (QFT) tubes. Evidence of cytokine production in response to QFT mitogen stimulation was observed in all four ELISA assays. However only the Mabtech Cat IFN-c ELISABasic kit could distinguish between M. bovis–infected (n = 1) and uninfected (n =1) cheetahs and was therefore selected for further evaluation. A preliminary cheetah specific cutoff value (11 pg/ml) for detecting M. bovis infection using the Mabtech Cat IFN-c release assay was calculated using a M. bovis uninfected cheetah cohort. **Although this study only included one confirmed M. bovis culture-positive and one M. bovis culture-negative cheetah, the Mabtech Cat IFN-c release assay demonstrated its potential for diagnostic application in this species.** Key Points: - As far of conservation efforts, translocations occur to help manage fragmented populations, and there is risk associated with movement b/t regions and countries= threat of introducing new pathogens, such as Mycobacterium bovis into naive pops - Wild felids can be infected with M. bovis from consuming infected or being in contact with infected animals - Currently the tuberculin skin test (TST) is only wildly available antemortem bTB test for wild felids, but poses logistical challenges when used in free-ranging animals since they have to be held for 72 hours and requires second immobilization– need other options - **Studies show that interferon gamma, tumour necrosis factor alpha, and interleukin 12 are important cytokines involved in host’s natural response to Mycobacterium tuberculosis infections**, and in domestic cats, these cytokines have been detected in mitogen-simulated and mycobacterial antigen-stimulated whole blood from M bovis-infected animals- diagnostic potential - This study supports the use of CRAs to measure immune activation in cheetahs, using plasma from QFT Mit-stimulated whole blood- however could not differentiate the antigen-specific TNF alpha and IL-1𝛃 conc from M. bovis cheetah vs uninfected cheetah - Previous studies show combined or parallel measurement of more than one cytokine can provide additional insights into disease state as opposed to measuring only one cytokine - Validation not possible - Proof-of-concept principle - goal of study - **The Mabtech Cat IFN-Ɣ ELISA kit, QFT stimulation platform, was able to detect immune activation and M. bovis sensitization in cheetahs, demonstrating its potential for diagnostic applications in cheetah and other felid species.**

Answer 92

J Zoo Wildl Med 2021 52(4):1224-1228, **DURATION OF ANTIGEN SHEDDING AND DEVELOPMENT OF ANTIBODY TITERS IN MALAYAN TIGERS (PANTHERA TIGRIS JACKSONI) NATURALLY INFECTED WITH SARS-CoV-2** Cushing AC, Sawatzki K, Heather NG, Puryear WB, Kelly N, et al. ABSTRACT: Natural infection of three captive Malayan tigers (Panthera tigris jacksoni) with SARS-CoV-2 caused mild to moderate symptoms of lethargy, anorexia, and coughing. Each tiger was longitudinally sampled opportunistically via consciously obtained oral, nasal, and/or fecal samples during and after resolution of clinical signs, until 2 wk of negative results were obtained. **Persistent shedding of SARS-CoV-2 genetic material was detected via reverse transcription–polymerase chain reaction in feces up to 29 d after initial onset of clinical signs, but not in nasal or oral samples.** Tigers became resistant to behavioral training to obtain nasal samples but tolerated longitudinal oral sampling. Serum was obtained from two tigers, and antibody titers revealed a robust antibody response within 9 d of onset of clinical signs, which was sustained for at least 3 mon. The tigers were infected despite the use of masks and gloves by husbandry personnel. No known cause of the outbreak was identified, despite extensive investigational efforts by the regional health department. No forward cross-species transmission was observed in primates housed in nearby enclosures. The increasing regularity of reports of SARS-CoV-2 infection in nondomestic felids warrants further investigations into shedding and immunity Study Design: Case series (n=3 individually housed adult Malayan tigers) Goal: - Document infection and protracted fecal shedding of SARS-CoV-2 in captive tigers - Suggest noninvasive fecal samples are a suitable for SARS-CoV-2 screening in captive nondomestic felids Key Points: - Nondomestic felid species are susceptible to natural infection and clinical signs associated with SARS-CoV-2 -- Clinical signs: coughing, lethargy, and anorexia resolving within ~2 weeks -- Anthropozoonotic infection has been documented from mink, but has not yet been documented in nondomestic felids - Tigers became less amenable to nasal swabs, oral swabs were more easily tolerated, and fecal samples were simple to collect - Fecal shedding of SARS-CoV-2 can be especially prolonged, though it is not clear if individuals are infectious during these periods -- Fecal RNA shedding persists for up to 35 d in lions -- Duration of tiger fecal shedding (up to 29) was less than in lions (35 d) - Fecal matter has an elevated risk of aerosolization during cleaning with high pressure hoses and could present a novel transmission route in zoological settings -- There was no evidence of infection in gibbons and gorillas, despite movement of staff between tiger and primate exhibits - It is unknown if the initial human-to-tiger transmission was via direct contact, indirect contact, or fomite -- The most likely source is those with direct tiger contact. However, all staff with direct or close contact wore masks and gloves -- One potential source was from a staff worker via preparation of diets; this staff person demonstrated respiratory signs 48 h before tigers showed signs -- This person tested negative via PCR within 7 d of clinical signs, and negative for antibodies during the Knox County investigation - The possibility of infection from members of the public remains viable TLDR: - Natural infection of 3 Malayan tigers with SARS-CoV-2 caused mild to moderate symptoms of lethargy, anorexia, and coughing - Persistent shedding of SARS-CoV-2 was detected in feces up to 29 d after onset of clinical signs - Fecal samples were simple to collect - Antibody titers revealed a robust response within 9 d of clinical signs, which lasted ~3 months - Tigers were infected despite staff wearing PPE; no known cause of the outbreak was identified

Answer 93

J Zoo Wildl Med 52(4), 1280-1285 **SUCCESSFUL SURGICAL CORRECTION OF PHIMOSIS IN AMUR LEOPARDS (PANTHERA PARDUS ORIENTALIS) USING A CARBON DIOXIDE LASER: CASE SERIES** Kane LP, Langan JN, Adkesson MJ ABSTRACT: **Three juvenile (<1 yr of age), genetically related Amur leopards (Panthera pardus orientalis) were diagnosed with phimosis.** In all cases, no clinical signs were identified, and phimosis was detected during routine examination. **Surgical enlargement of the preputial orifice was performed successfully using carbon dioxide laser with a 0.25-mm tip and an 8-watt continuous wave setting.** A linear incision on the ventral aspect of the preputial opening allowed for complete preputial extrusion followed by placement of simple-interrupted sutures on both sides of the incision. No postoperative complications were observed, and there was no evidence of phimosis recurrence at subsequent examinations. Preputioplasty with carbon dioxide laser was used to successfully manage Amur leopards with phimosis. Given relatedness of these cases, a genetic predisposition for phimosis development in Amur leopards must be considered. Goal: Describe phimosis in three 2-10 mo, male, related Amur leopards as well as successful laser preputioplasty Key Points: - Amur leopards (Panthera pardus orientalis) are 1 of 9 extant subspecies of the common leopard - They’re found only in SE Russia and northern China, with < 100 free-ranging individuals - Phimosis = inability to extrude penis from prepuce, occurs 2° to a preputial orifice narrowing - In domestic felids, it is described as congenital or acquired due to trauma, excessive grooming, or complications from a lower UTI - Phimosis can occur shortly after birth due to excessive perineal cleaning by the dam or aberrant prepuce suckling behavior from littermates - Animals may be asymptomatic or have pollakiuria, stranguria, urine dribbling, excessive preputial licking, preputial distention secondary to urine retention, and vocalization - Phimosis in these cases is presumed to be congenital - Preputial trauma at an early age from dam or littermates cannot be excluded, but no such behavior was observed directly or via nest box cameras TLDR: - Apparent congenital phimosis in Amur leopards represents a previously unrecognized reproductive concern for this critically endangered subspecies - Laser preputioplasty in all three cases was curative - It provides an excellent prognosis, low potential for complications, and successful reproduction post-operatively

Answer 94

**PERFORMANCE OF THE TUBERCULIN SKIN TEST IN MYCOBACTERIUM BOVIS–EXPOSED AND –UNEXPOSED AFRICAN LIONS (PANTHERA LEO) ** Viljoen et al. Journal of Wildlife Diseases, 55(3), 2019, pp. 537–543 ABSTRACT: Lion (Panthera leo) populations, classified as vulnerable under the International Union for Conservation of Nature red list of threatened species, are facing a variety of threats, including tuberculosis (TB) caused by Mycobacterium bovis. The lack of knowledge on pathogenesis and diagnosis of TB, the prolonged course of the disease, the existence of subclinical infection, and nonspecific clinical signs hamper management of TB in both free-ranging and captive lion populations. Early and accurate antemortem diagnosis of M. bovis infections is important for disease management. In this study, we **investigate the suitability of the single intradermal cervical test (SICT), developed with free-ranging Kruger National Park (KNP) lions exposed to M. bovis, for use in other lion populations. Using the recommended interpretation, the specificity of the SICT was low in disease-free captive lions, leading to false-positive diagnoses in 54% of individuals in the present study. Alternative interpretations of the tuberculin skin test are proposed that significantly reduce false-positive diagnosis in the sampled captive lions without significantly affecting diagnoses in the KNP lions**; these changes may facilitate screening for M. bovis infection regardless of the exposure status of the lion population being investigated. Tuberculosis (TB) due to Mycobacterium bovis may have impact on lion populations, leading to subclinical infections, chronic disease with non-specific signs, and death in advanced stages of disease - **Gold standard test = mycobacterial culture post-mortem** Antemortem tests in lions: - Tracheobronchial lavage (BAL) sample for culture - Detection of antigen-specific immune responses - **Single intradermal cervical test (SICT)**- Keet et al. (2010) reported high specificity (81%) and sensitivity (87%) in infected lions in KNP with a cut-off of PPDb skin thickness response > 2 mm -- This cut-off may not be good for testing lions in other settings - **Single intradermal comparative cervical test (SICCT) is a skin test with injection of both bovine and avian purified protein derivatives (PPDs) and measurement of changes of skin fold thickness at each site - Accounts for cross reactivity to nontuberculosis mycobacteria (NTM)** Study details: Aim is to investigate the performance of TST in different populations of 36 wild (exposed) and 26 captive (control) lions to calculate cut-off values for other populations distinct from original KNP group: - SICT using previous cut-off values showed 78% of wild and 28% captive lions were test positive -- Suggests low specificity (likely false positives) since captive animals were never exposed - **With the “sensitive” interpretation of SICCT 12% captive animals were test positive, but with “specific” interpretation of SICCT no captive animals were positive: -- Hence, the specific interpretation (using PPDb >2 mm and 2 mm greater than PPDa) of SICCT is recommended to reduce false-positive results** - Response to PPDa injection site of captive lions was greater than PPDb and compared to wild lions -- Suggests greater exposure to NTM than wild lions - BAL cultures and antigen-specific gene expression assays (QFT-GEA) were used for comparison of TST -- BAL was insensitive for exposed lions (only one tested positive) -- QFT-GEA was sensitive - high proportion of exposed and low proportion of unexposed lions positive -- Likely detecting antigen-specific immune sensitization in KNP lions Take-away: - Diagnostic interpretation of SICT is appropriate for diagnosis of M. bovis infection in KNP lions, but lacks specificity for lions from other populations and alternative interpretations to reduce false positives are proposed. - Recommend that the specific interpretation of SICCT to be used when screening lions for M. bovis infection, especially in low prevalence populations.

Answer 95

**TIGER (PANTHERA TIGRIS) AND DOMESTIC CAT (FELIS CATUS) IMMUNE RESPONSES TO CANARYPOX-VECTORED CANINE DISTEMPER VACCINATION** Michael McEntire, Edward C. Ramsay, Stephen Kania, Peter Prestia, Eman Anis, Andrew C. Cushing, Rebecca P. Wilkes; JZWM 50(4) Abstract **Two methods for delivering a canarypox-vectored canine distemper vaccine to tigers (Panthera tigris) and domestic cats (Felis catus) were investigated. Eight tigers were divided randomly into two vaccination groups: subcutaneous injection or topical tonsillar application.** Each tiger received 2 ml of canine distemper virus (CDV) vaccine (Merial Ferret Distemper Vaccine). Blood was collected from tigers on days 0, 21, 35 or 37, and 112 post–initial vaccination (PIV). **Domestic cats were divided randomly into four treatment groups: saline injection (negative controls), low- and high-dose oral, and subcutaneous vaccinates.** Blood was collected from domestic cats on days 0, 7, 21, and 28 and 165 or 208 PIV. Sera were tested for CDV antibodies by virus neutralization. All individuals were seronegative at the beginning of the study. *One tiger vaccinated subcutaneously developed a titer of 32 by day 35, which reduced to 16 by day 112. Another tiger vaccinated by tonsillar application developed a titer of 8 on day 112*. All other tigers remained seronegative. Cats that received saline injection or oral vaccination remained seronegative at each sampling time. *Domestic cats vaccinated subcutaneously developed titers ranging from 4 to >128 by day 28, and those re-bled at day 166 had titers of 16 or 64.* **The disparity in response between domestic cats and tigers may be due to species differences or it may represent a dose-dependent effect. Subcutaneous vaccination with canarypox-vectored Purevax Ferret Distemper® is safe and elicits persistent antibody titers in domestic cats vaccinated parenterally.** Key Points: - Identification of CDV infection in wild Amur tigers (Panthera tigris tigris), a species listed as endangered by the International Union for the Conservation of Nature, poses an important conservation threat to this species' endangered population - Vaccination of tigers with a canarypox-vectored recombinant canine distemper vaccine has not caused vaccine-induced illness, which has previously been seen with live attenuated distemper vaccines. -- However, canarypox-vectored vaccines have been unable to produce a humoral response sufficient to be protective - A live attenuated CDV vaccine produced a profound antibody response in tigers, but a single pregnant tiger vaccinated with that product produced a litter of kittens with congenital heart defect - The signaling lymphocyte activation molecule (SLAM) that has been identified as a receptor for CDV is highly conserved across different felid species - This study was performed to determine the safety and efficacy of a monovalent canarypox-vectored recombinant vaccine administered parenterally and orally to tigers. Domestic cats were evaluated to determine if they would be a suitable model for future CDV vaccination studies - All tigers had negative titers against CDV prior to vaccination, and only two tigers developed titers following vaccination - All domestic cats had negative titers against CDV prior to vaccination, and all cats that received PFD orally or saline continued to have negative antibody titers at all sampling times - All cats vaccinated SC developed titers, ranging from 4 to >128 (geometric group mean = 34.30), on day 28. - The larger serologic responses in parenterally vaccinated domestic cats compared to the responses in the subcutaneously vaccinated tigers suggest tigers may be a dose-dependent effect to this vaccine, as the domestic cats received approximately 10 times the quantity of vaccine/kg as the tigers - Further work is needed to determine if a higher dose of PFD will produce sufficient humoral and cell-mediated responses to suggest protection

Answer 96

DENTAL DISEASES AND OTHER ORAL PATHOLOGIES OF CAPTIVE JAGUARS (PANTHERA ONCA) FROM BELIZE, CENTRAL AMERICA - Lindsey A. Schneider et al. Journal of Zoo and Wildlife Medicine 51(4): 856–867, 2020 Abstract: Dental and oral diseases are prevalent in many mammalian species including wild felids. Determining the dental and oral health status of captive animal populations can help establish preventive and therapeutic strategies, leading to improved welfare and conservation efforts. The aim of this study was to assess the prevalence of periodontal disease, endodontic disease, tooth resorption, and other clinically relevant dental and maxillofacial abnormalities in a population of captive jaguars (Panthera onca) using clinical, radiographic, and histopathological findings. Fifteen jaguars, ranging from young adult to geriatric, kept at a private zoo in Belize, Central America, had a detailed oral examination under general anesthesia between January 2015 and March 2019. **Periodontitis was present in 3.8% (16/423) of examined teeth and 53.8% (7/13)** of jaguars that underwent periodontal probing. **Endodontic disease secondary to dentoalveolar trauma was found in 21.0% (89/423) of teeth in 73.3% (11/15) of animals. Tooth resorption, which has not been previously documented in jaguars, affected 1.4% (6/423) of teeth in 13.3% (2/15) of jaguars**. Other abnormalities included metallic foreign material (gunshot) identified radiographically in 33.3% (5/15) of jaguars and nontraumatizing malocclusion in 9.1% (1/11) of jaguars that had occlusion evaluated. **Much of the oral pathology identified in captive jaguars is suspected to arise from capture and/or captivity-associated behaviors, as suggested by gunshot around the oral cavity, fractures of rostral teeth (canine and incisor teeth), and abrasions consistent with cage-biting on canine teeth**. Anesthetized oral examination—including full-mouth intraoral radiographs, periodontal probing, and charting—is recommended for jaguars with clinical signs of oral pain, as well as for routine systemic evaluation. Jaguar (Panthera onca) dental formula: I3/3; C1/1; P3/2; M1/1 per side = 30 teeth - Periodontal disease, endodontic disease, and tooth resorption are significant problems that contribute to morbidity in captive jaguars Study details: - Endodontic disease secondary to trauma was the most common pathology in this study (particularly fractured canine and incisors and abrasions to distal aspects of canine teeth) 🡪 suspect due to captive behavior -- High failure rate of root canal therapy (50%) in this population - 13 animals had periodontal probing – periodontitis in ~54% - High frequency of gingival recession at canine teeth with distance between the cemento-enamel junction (CEJ) gingival margins is >1 mm (22/50 probed; 61.5% of animals) -- Also seen in captive tigers – may normal anatomy vs. represents extrusion of tooth vs. alveolar bone loss -- Probing depths of >3mm were considered attachment loss - Pathologic tooth resorption in 13% (2/15 animals) – reported in other nondomestic felids - Significant bacteria noted associated with lesion - Malocclusion and developmental dental abnormalities infrequently observed - Two individuals missing left maxillary second premolar tooth -- Second maxillary premolars have been suggested to be disappearing from wild felids over course of evolution (present in mountain lion, but lost in bobcat) - Age, diet and other environmental factors such as frequency of periodontal treatment should be considered when assessing risk of periodontal disease in captive felids - Endodontic disease secondary to trauma appears to be related to captive behaviors -- Previous study found minimal endodontal disease in wild felids Take-away: Anesthetized oral exam including intraoral rads, periodontal probing and charting is recommended for routine systemic evaluation of jaguars.

Answer 97

**Comparison of blood symmetric dimethylarginine and creatinine as endogenous markers of kidney function in captive tigers** JZWM, 52(2):628-637 Abstract: Chronic kidney disease (CKD) is a common cause of morbidity and mortality in captive tigers (Panthera tigris). Blood creatinine (Cr) and blood urea nitrogen measurements are inexpensive and common biomarkers used to evaluate renal function. However, several limitations have been reported regarding their sensitivity and interindividual variability. **Symmetric dimethylarginine (SDMA) has been suggested to be a more sensitive biomarker that is less affected by extrarenal factors and has a strong correlation with glomerular filtration rate and blood Cr in several species.** This project aimed to identify the usefulness of SDMA as an endogenous marker of kidney function in captive tigers. The hypothesis of this study is that increased circulating SDMA is positively associated with increased blood Cr. SDMA and Cr were measured in 65 banked samples (serum and plasma) from 30 individual captive tigers. The samples were collected over a 38-y period and stored at =21C. SDMA and Cr concentrations were determined using the commercially available SDMA test and enzymatic colorimetric methods, respectively. **SDMA had a significant positive association with Cr (for every 1 unit increase of log SDMA, Cr increased by 82%, P = 0.0002). Age and subspecies influenced Cr but not SDMA concentrations.** In one animal, blood SDMA increased above the ZIMS reported range. approximately 3.6 mo before Cr increased. SDMAis currently indicated for the diagnosis of CKD in domestic felids and seems also promising in nondomestic felids. Further prospective studies might improve the understanding of the performance of this biomarker. Basics - The most common pathological findings reported in tigers with renal disease include interstitial nephritis, pyelitis, intratubular concrements, and tubular degeneration. - Creatinine (Cr) and blood urea nitrogen (BUN) are inexpensive and common biomarkers used to evaluate kidney function. - In the early stages of kidney injury, significant changes in GFR result in minimally detectable alterations in blood Cr. - In advanced kidney disease small changes in GFR have a significant effect on blood Cr concentration. -- Extrarenal factors, such as endogenous production by muscle, dietary protein intake, age, and sex, can affect blood Cr levels. -- BUN is also affected by extrarenal factors, such as dietary protein intake, dehydration, reduced cardiac output, and other pathologies (e.g., gastrointestinal hemorrhage, pyrexia). - SDMA is a product of intracellular protein methylation and appears to be eliminated primarily by the kidney. -- SDMA is more sensitive to decline in GFR and allows for diagnosis of kidney disease earlier. -- SDMA may be more clinically reliable and specific than Cr because it appears to be less affected by extrarenal factors. - Plasma and serum concentrations of SDMA were highly correlated with GFR (in humans, domestic dogs and cats, and cheetahs. Details: - Age and subspecies influenced Cr but not SDMA concentrations. -- Cr increased by 4% each year as age increased. -- Sumatran tigers demonstrated 41% higher blood Cr concentration than Amur tiger Cr. -- Neither age nor subspecies was associated with SDMA concentration. - Historical Cr results did not change substantially over time when compared with recent Cr results. - SDMA had a significant positive association with Cr (for every 1 unit increase in log SDMA, Cr increased 82%). - In one of seven tigers, blood SDMA increased above the ZIMS range approximately 3.6 mo before an increase in Cr. - The 65 measured blood SDMA and Cr results ranged from 5 to 56 lg/dl and from 0.5 to 11.4mg/dl. Take Home: This study indicates that SDMA should be indicated for use as a supplemental tool in the diagnosis of CKD in domestic felids and seems also promising in nondomestic felids. This is the first study of SDMA in captive tigers.

Answer 98

**Keratometry, biometry, and prediction of intraocular lens power in adult tigers (Panthera tigris)** Cameron D. Owens, Tammy M. Michau, Justin Boorstein, Elizabeth R. Wynn, and Richard J. McMullen Jr Taxonomy: Mammalia→ Carnivora→ Felidae→ Panthera tigris Abstract: OBJECTIVE To calculate the necessary pseudophakic intraocular lens (IOL) power to approximate emmetropia in adult tigers. ANIMALS 17 clinically normal adult tigers. PROCEDURES 33 eyes of 17 clinically normal adult tigers underwent routine ophthalmic examination and B-scan ultrasonography while anesthetized for unrelated procedures. Specific ultrasound data (globe measurements and corneal curvature) and estimated postoperative IOL positions were utilized to calculate predicted IOL power by use of Retzlaff and Binkhorst theoretical formulas. Applanation tonometry and refraction were also performed. RESULTS Mean ± SD axial globe length was 29.36 ± 0.82 mm, preoperative anterior chamber depth was 7.00 ± 0.74 mm, and crystalline lens thickness was 8.72 ± 0.56 mm. Mean net refractive error (n = 33 eyes) was +0.27 ± 0.30 diopters (D). By use of the Retzlaff formula, mean predicted IOL power for the postoperative anterior chamber depth (PACD), PACD – 2 mm, and PACD + 2 mm was 43.72 ± 4.84 D, 37.62 ± 4.19 D, and 51.57 ± 5.72 D, respectively. By use of the Binkhorst equation, these values were 45.11 ± 4.91 D, 38.84 ± 4.25 D, and 53.18 ± 5.81 D, respectively. **Mean intraocular pressure for all eyes was 14.7 ± 2.69 mm Hg.** CLINICAL RELEVANCE The calculated tiger IOL was lower than reported values for adult domestic felids. Further studies evaluating actual PACD and pseudophakic refraction would help determine the appropriate IOL power to achieve emmetropia in this species. Key Points: - **Cataracts inherited or acquired, causes of acquired include nutritional deficiencies, certain toxins, trauma, inflammation, endocrine disorders (eg DM) and senility** - **Deficiencies in taurine, arginine, phenylalanine, tryptophan and histidine are possible causes of development of cataracts and strabismus in tiger cubs fed particular artificial milk replacers** - There are now more tigers in managed care than the wild - Gold standard of cataract treatment is surgical removal of affected lens, removal of lens without placement of artificial lens leaves the eyes severely hyperopic (farsighted) due to role of the lens in optical refraction. - Objective was determine the mean intraocular pressure obtained via applanation tonometry, mean phakic refraction and mean ocular biometric data of tiger eye (axial globe length AGL, anterior chamber depth ACD, crystalline lens thickness CLT and corneal curvature to calculate an estimated pseudophakic IOL power to achieve emmetropia in the tiger Discussion: - Tigers had greeted AGL than domestic cats and dogs, but shorter AGL than horses - Mean ACD was greater in tigers than in cats, dogs and horses - Mean corneal curvature was less than in domestic cats and dogs, but greater than in horses. - Corneal curvature has an important and direct effect on the strength of an IOL necessary to refract light into the retina - No significant difference in ocular dimensions based on weight, age, and sex. - Calculations in present study suggest tigers may required a higher IOL power than has been reported for other species but lower power requirement than reported for domestic cats for achieving emmetria- not similar to cats - Tigers and cats have similar lens thicknesses, but other variables such as the PACD and AGL and corneal curvatures were very different - Tigers lens rests more anteriorly than domestic cats - Yet to be determined whether these formulas are appropriate for estimation of IOL power in tigers - The Retzlaff formula consistently produced an IOL power lower than did the Binkhorst formula - Concern for overcorrection resulting in myopia can occur with these formulas - Based on the formula used, would need to place more anteriorly or most posteriorly. - **IOP’s can be affected by anesthesia, ketamine can spuriously increase, alpha-2’s can decrease. However this is representative of how would read in tigers clinically as dangerous to do awake.**

Answer 99

**Complications associated with total hip arthroplasty in four large nondomestic felids** AJVR 2022.1: Lauren P. Kane, multiple DACZM CASE DESCRIPTION A 9-year-old 37-kg sexually intact male snow leopard (Panthera uncia) with no history of lameness but radiographic evidence of right femoral subluxation and flattening of both femoral heads, 2 juvenile (< 1 year old) 25-kg sexually intact male cheetahs (Acinoynx jubatus) with unilateral hind limb lameness resulting from trauma, and an 11-yearold 110-kg sexually intact female Amur tiger (Panthera tigris altaica) with a 2-year history of left hip joint osteoarthritis were examined. CLINICAL FINDINGS No clinically relevant clinical findings other than hip joint problems were identified. All 4 felids underwent staged bilateral (snow leopard) or unilateral (cheetahs and tiger [Panthera tigris]) total hip arthroplasty (THA). TREATMENT AND OUTCOME In the snow leopard, both femoral THA components were found to be luxated 1 year after surgery. Treatment consisted of autogenous corticocancellous rib graft augmentation of the dorsal acetabular rims and synthetic suture capsulorrhaphies. The snow leopard lived for an additional 4 years with no additional THA-related complications. In the other 3 animals, catastrophic complications (luxation in the cheetahs and femoral fracture in the tiger) occurred shortly after THA. The THA implants were removed, and excision arthroplasty was performed. Long-term outcomes were good in all 3. CLINICAL RELEVANCE Findings underscore the challenges associated with THA in large nondomestic felids. Given the high risk for early catastrophic failure as a result of luxation or fracture, plans must be made and resources must be available in case revision surgery or implant removal with excision arthroplasty becomes necessary. Key points: - DJD is the progressive destruction of 1 or more joint component -- **Hip joints most commonly affected appendicular joints in felids** -- Best described in snow leopards, but also cheetahs and tigers -- Surgical techniques: THA - goal to mitigate pain while improving hind limb function -- Pelvic and femoral osteotomy -- Femoral head & neck ostectomy – excisional arthroplasty to relieve pain secondary to abnormal bony contact in end-stage DJD or salvage after failed THA This report describes 4 large felids that underwent THA for treatment of hip DJD secondary to presumed hip dysplasia, avascular necrosis, trauma, and primary osteoarthritis: - All THA’s failed; including a custom-designed, patient specific THA implant for the Amur tiger - **Domestic cats apply greater peak vertical force in the forelimbs vs. Large nondomestic felids that apply greater forces in their hind limbs.** - **Large felids have heavy hind limb bones, a relatively small volume of muscle in the hip region, large psoas muscles, and extreme hip joint ranges of motion** - Difficult to control post-op activities - THA instrumentation, implants, and surgical techniques must account for anatomic and biomechanical differences between large nondomestic and domestic felids - Implant removal and excision arthroplasty salvage procedures resulted in acceptable outcomes - **Peri-op analgesia varied: Peri-incisional liposomal bupivacaine, morphine epidermal, fentanyl transdermal patch, NSAIDs, opiates, gabapentin, amantadine and anxiolytics (Trazadone)** - **Opioids have been the mainstay of periop pain management in companion animals** - dampen afferent nociceptive transmission in PNS and CNS. - Tramadol, a synthetic opioid, increases activity and quality of life in geriatric cats w OA

Answer 100

**YERSINIA PSEUDOTUBERCULOSIS INFECTION IN LIONS (PANTHERA LEO) AT A ZOOLOGICAL PARK.** Cabot ML, Watanabe TT, Womble M, Harrison TM. Journal of Zoo and Wildlife Medicine. 2022;53(3):593-599. Two co-housed 17-yr-old male lions (Panthera leo) and one 15-yr-old female lion in an adjacent enclosure developed acute lethargy, depression, anorexia, and ataxia at a zoological park in central North Carolina, United States. One of the male lions and the female lion were found dead 5 and 4 d after the onset of clinical signs, respectively. The other male lion recovered without any clinical treatment. A third male lion housed with the female lion never developed clinical signs. Postmortem examination, microscopic evaluation, and bacterial culture of the liver from both deceased lions confirmed systemic Yersinia pseudotuberculosis infection. Susceptibility testing revealed resistance to amoxicillin–clavulanic acid and cefazolin. To investigate the extent of the outbreak, qualitative real-time polymerase chain reaction (PCR) for Y. pseudotuberculosis was performed on feces and substrate from 15 enclosures housing nondomestic felids and canids, resulting in a positive sample from one enclosure housing four asymptomatic lions. This enclosure was adjacent to housing of the deceased female lion. Enrofloxacin was administered to all animals in the vicinity at doses of 5 mg/kg in nondomestic felids and 10 mg/kg in nondomestic canids, orally q24h for 14 d. Repeated fecal PCR performed 1 wk after completing the antibiotic treatment protocol found no positive samples. The source of the infection was not identified despite PCR testing of environmental samples from all enclosures in the vicinity, the remains of a prey item fed out prior to the outbreak, and a single dead rodent found on grounds. No further clinical cases have occurred within the following year. Background - Yersinia pseudotuberculosis - gram negative facultative anaerobic coccobacillus, family Enterobacteriaceae - ZOONOTIC, ubiquitous Key Points - 3 clinical, 2 mortalities, 4 asymptomatic positive cases in lions - Clinical and morts were geriatric - After extended cold weather and rain - psychrophilic, opportunistic bacteria - CS - lethargy, anorexia, nausea, abdominal discomfort, ataxia/hind-end weakness - BW - mild lymphocytosis, hypokalemia, hypochloremia, high ALT, high tbili - Gross - severe, diffuse interstitial pneumonia, multifocal, embolic, suppurative hepatitis - Histo - Microabscesses in liver, lungs, spleen, kidney; myriad gram-negative coccobacilli - Culture: Y pseudotuberculosis, pan-susceptible or resistant to Clavamox and cefazolin - Positive fecal PCR in asymptomatic animals - Treated with enrofloxacin - no further cases or positives - Unable to find source - negative PCR of substrate/water, shared raw prey item, and dead rat on property Conclusions - Lions are susceptible to Y pseudotuberculosis - causes disseminated abscesses (‘pseudotuberculous lesions’) especially in the liver and spleen with rapid clinical course and mortality - May be asymptomatic carriers/shedders - May require fluoroquinolones to treat and stop mortalities - Fecal PCR is a viable screening tool

Answer 101

Journal of Zoo and Wildlife Medicine 53(4): 744–754, 2022 **DETERMINATION OF SPECIFIC ENTEROPATHOGEN PRESENCE IN CAPTIVE CHEETAHS (ACINONYX JUBATUS) FED VARIOUS DIETS USING FLUORESCENCE IN SITU HYBRIDIZATION** Melanie Peel, DVM, Rodrigo Garces Torres, DVM, EMCV (FS), Benjamın Alcantar Hernandez, MVZ, Curtis Eng, DVM, and Brian B. Oakley, PhD– Rev by AJC Abstract: Chronic enteropathies pose an important difficulty in the captive management of cheetahs (Acinonyx jubatus) because of suspected multifactorial pathogenesis and the complex nature of enteric microbiota dynamics. **Enterobacteriaceae, Campylobacter spp., Clostridium perfringens, Helicobacter spp., and Salmonella spp. are enteropathogens of interest because of their zoonotic potential and suspected contribution to enteropathies**. This study aimed to determine the presence of these enteropathogens of interest in fecal samples from cheetahs (N = 48) fed different diets from three different institutions and to investigate the associations between diet, fecal score, and specific enteropathogen presence. Fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes were used to visualize and quantify putative enteropathogens in each sample concurrent with selective culturing for Salmonella and Clostridium perfringens. From FISH counts, carcass-fed animals had greater numbers of Enterobacteriaceae compared with animals fed low-fat dog food, although this trend was not statistically significant (P = 0.088). Furthermore, no significant associations were found between fecal score and bacterial load. **Abundance of Campylobacter spp., Clostridium perfringens, or Helicobacter spp. as measured by FISH were not correlated with diet or fecal score**. On the basis of these data, in agreement with published literature, it is concluded that **these microbes may be commensals in the cheetah gastrointestinal tract** and do not appear to be a primary cause of abnormal fecal scores. Key Points: - Chronic gastritis can be associated with Helicobacter spp though disease is multifactorial - Many presumed enteropathogens may be normal commensals - More enteropathogens were isolated from managed cheetahs than free ranging (C. perfringens most common) - Objective: assess for presence of enteropathogens in cheetahs with different diets and correlate this to fecal scores - FISH diagnostic determined to be a reliable diagnostic for entropathogens in this study - Abundance of specific bacteria and Enterobacteriaceae was not significantly different by fecal score or diet - Enterobacteriaceae increased in carcass fed cheetahs (not significantly different) - Helicobacter spp., C, perfringens, and Campylobacter spp. found in the majority of samples and were not correlated with fecal score or diet. Salmonella not detected. - These bacteria should be considered commensals/opportunistic pathogens - Carcass fed animals had a significantly higher prevalence of soft feces Take Home: FISH was a reliable method to detect Helicobacter, C. perfringens, and Campylobacter in healthy, and there was no correlation to diet or fecal score.

Answer 102

Journal of the American Veterinary Medical Association, 260(13), 18. (2022). **Blood concentration of symmetric dimethylarginine correlates with kidney damage as assessed with a proposed histologic grading system for chronic kidney disease in tigers (Panthera tigris)**. Andrews, B. J., Cushing, A. C., Murphy, R. E., Wilson, E. M., & Sula, M. M. – reviewed by ZCR Abstract OBJECTIVE To determine the utility of blood symmetric dimethylarginine (SDMA) concentration measurement as a diagnostic tool for chronic kidney disease (CKD) in tigers (Panthera tigris) by comparing results for SDMA with those for traditional renal biomarkers and investigating correlations between these biomarkers and histopathologic kidney changes in tigers with CKD. SAMPLE Blood, urine, and kidney samples from 35 tigers with CKD from 2 sanctuaries. PROCEDURES Blood (serum or plasma) and urine samples were collected antemortem. Necropsy, including gross and histologic assessment, was performed for tigers that died or were euthanized for quality-of-life reasons. Results for CKD biomarkers in blood (BUN, creatinine, phosphorus, and SDMA concentrations) and urine (protein concentration, urine protein-to-creatinine ratio, and urine specific gravity) were evaluated for correlation with histologic kidney damage scored with an objective grading scale defined by percentage of inflammation, fibrosis, and tubular atrophy. RESULTS **Symmetric dimethylarginine had the strongest significant correlation (ρ = 0.667) with histologic kidney damage score, followed by urine specific gravity (ρ = –0.639), blood creatinine concentration (ρ = 0.624), and BUN (ρ = 0.588)**. No significant correlation with kidney score was identified for blood phosphorus concentration, urine protein concentration, or the urine protein-to-creatinine ratio. CLINICAL RELEVANCE We recommend SDMA be prioritized as a renal biomarker in tigers, with SDMA results considered in addition to those of other traditional renal biomarkers when assessing kidney function in tigers. Additionally, the grading scale we developed could be replicated across patients and pathologists for more consistent postmortem assessment of CKD in tigers. Key Points - Renal disease historically diagnosed via high BUN, Cr, P or UPC alone or in combination with UA findings - Symmetric dimethylarginine (SDMA) is a product of protein metabolism, product of methylation of arginine protein residues, > 90% of SDMA excreted by kidneys -- Showing promise in other species – marker of 25-40% decrease of GFR in domestic cats, versus 75% for creatinine -- Validated in domestic cats and cheetahs -- SDMA also has less extrarenal influences. Cr influenced by muscle mass, and BUN influenced by hepatic function and nutritional status. - MC histologic renal lesion in tigers reported is interstitial nephritis in 36% of tigers M&M - 2016-2020: plasma and serum, urine collected from 86 tigers at Tiger Haven and In-Sync Exotics for renal biomarker analysis - Necropsy on 33 tigers: histologic grading scale determined based off of tubulointerstitial nephritis, manifested as interstitial inflammation, fibrosis and tubular atrophy. Scores defined as mild, moderate or severe. - Age range 7-35 years - SDMA was the biomarker with the greatest correlation to histologic kidney score, followed by creatinine, BUN, and USG (negative correlation). Blood P, UPC and urine protein concentration did not. - SDMA increase correlated with all 3 categories of histologic changes: inflammation, fibrosis and tubular atrophy Take home message: SDMA is correlated more strongly with histologic kidney score than other renal biomarkers measured, indicating that it should serve as a reliable way to identify renal disease earlier on antemortem.

Answer 103

Zoo Biology. 2022;41:328–339 **Dietary lutein supplementation improves immunity and antioxidant status of captive Indian leopards (Panthera fusca)** Sharad M. Durge | Asit Das | Subodh K. Saha | Amit Pande | Dimpal Thakuria | Ankur Saxena | Yogesh Bhardvaj | Ashok K. Verma – rev by AJC Abstract: Indian leopards kept in zoos are fed solely on carabeef on bone (CBB) diets. Carabeef contains lesser or no carotenoids. Hence, the **captive Indian leopard diets are suspected to be deficient in carotenoids while their wild counterparts acquire these pigments from their natural prey**. Lutein is a vital carotenoid that plays its role as an antioxidant and immunomodulator. This experiment investigates the effect of lutein supplementation on antioxidant status, immunity, and stress in captive Panthera fusca fed CBB diets. Nine leopards were used based on 3 × 3 replicated Latin square designs in the experiment. Groups CON, LUT20, and LUT40 were supplemented with 0, 20, and 40 ppm of lutein, respectively. Each experiment comprised of 10 days of wash‐out period, 11 days of adaptation, and 4 days of collection. **Digestibility of crude protein (CP) was higher (p < .01) in groups LUT20 and LUT40. Serum concentration of protein, globulin, urea (p < .05), total carotenoids, total antioxidant capacity (TAC), catalase (CAT) activity, and lymphocyte transformation test (LTT) index were higher (p < .001) in groups LUT20 and LUT40**. Activity of superoxide dismutase (SOD) and serum concentration of immunoglobulin were higher (p < .001) in group LUT20. Serum concentration of malonaldehyde (MDA) and fecal concentration of cortisol decreased (p < .001) in groups LUT20 and LUT40. Serum concentration of total immunoglobulin (μg/ml) and LTT were higher in group LUT20. Fecal concentration of cortisol (ng/g) was lower in LUT20 and LUT40. T**he study concludes that supplementation of lutein at 20 ppm would improve antioxidant status and immunity and alleviate stress in captive Indian leopards.** Key Points: - Mammals lack the ability of de novo carotenoid synthesis - Ungulates, rodents, primates and birds accumulate carotenoids naturally through vegetative feeding - Lutein associated with anti-inflammatory, neuroprotective and antiangiogenic activities - Felids are good accumulators of carotenoids in their bodies - Lutein acts as an antioxidant by trapping free radicals and quenching singlet oxygen - Zoo diet of Indian leopards typically contains carabeef on bone – minimal carotenoids - Digestibility of crude protein and organic matter higher in groups with lutein (LUT20 and LUT 40) - Intake and absorption of carotenoids increased in LUT20 and LUT 40 (higher in LUT40). - Intake and absorption of Ca and P not different - Serum concentration of total carotenoids increased with increased carotenoid content - Concentration of serum Igs higher in LUT20 than LUT40 despite higher concentration of lutein in LUT40, thus supplementation at a dose > 20 ppm may not be beneficial - Reduced lipid peroxidation due to lutein supplementation in LUT20 and LUT40 - Total protein and urea higher in LUT20 and LUT40, otherwise no other differences in CBC/Chem values. - Supplementation of lutein promoted protein digestion - Increased globulins in LUT20 and LUT40 - Lutein did not affect the palatability of the diet - Intake of DM, organic matter, CP and gross energy not influenced by lutein supplementation - Supplementation of lutein decreased the fecal cortisol concentration Take-Home Message: - The digestibility of crude protein, serum concentration of total protein and globulin were higher in lutein supplemented groups than controls. - There was increased concentrations of serum carotenoids and total antioxidant activity in a dose dependent manner and fecal cortisol decreased with lutein supplementation. - Lutein increased serum concentration of immunoglobulin though no additional benefit seen using a higher lutein concentration (40 vs 20 mg/kg DM). - Supplementing lutein at higher levels may act as a prooxidant thus recommended 20 mg/kg DM.

Answer 104

Journal of Zoo and Wildlife Medicine, 53(4) : 844-854 2023 **MELANOCYTIC NEOPLASIA IN PANTHERA SPECIES: CLINICAL PRESENTATIONS, PATHOLOGIC FINDINGS AND RESPONSES TO TREATMENT** Ready, Zachary C., LoBato, Denae, LaDouceur, Elise, Garner, Michael M., and Cushing, Andrew C. Abstract: Neoplasia is a common cause of morbidity and mortality in captive nondomestic felids. Seven tigers (Panthera tigris), two African lions (Panthera leo), and two snow leopards (Panthera uncia) were diagnosed with melanocytic neoplasia (10 malignant melanomas, two benign melanocytomas) over a 20-yr period. **Animals were 10–19 yr old and 5/7 tigers were phenotypically white. **Malignant melanoma tumor location included skin (n=4), oral mucosa (n = 2), nasal planum (n = 1), iris/uvea (n = 2), and lip margin (n = 1); melanocytomas were found in skin (n = 2). Metastasis to regional lymph nodes was seen at diagnosis in 3/7 melanoma cases. Thoracic radiography (n = 6) and/or computed tomography (n = 2) did not detect pulmonary metastasis at diagnosis but were useful for detection later in the disease course. Median survival time (MST) for all cases ranged from 1 mon – 40 mon. Seven cases with malignant melanoma underwent treatment, which included surgery, radiation therapy, and administration of the canine melanoma vaccine (Onceptt) or a combination of these treatments; MST was 5– 40 mon for these cases. **While multimodal therapy may provide an improved survival time, the majority of animals with malignant melanoma invariably died from neoplastic disease**. Necropsy confirmed metastasis of malignant melanoma in 7/9 animals; sites included lung, liver, lymph node, kidney, mesentery, pleural cavity, heart, stomach, spleen, and adrenal gland. This case series describes the clinical and histologic findings of melanocytic neoplasia in nondomestic felids as well as multimodal treatment strategies incorporating the canine melanoma vaccine. Key points: - Previous neoplasia in Panthera -- Came most frequently from reproductive, endocrine/neuroendocrine, integument systems -- Mammary adenocarcinoma is the most common neoplasm -- Integumentary neoplasms are the 3rd most common with melanocytic neoplasia previously reported in tigers and lions - Melanocytic neoplasms -- Benign – melanocytoma -- Malignant – melanoma -- Both locally invasive and can metastasize to lymph nodes and lungs -- Treatment is primarily surgery, combined with radiation therapy -- Adding radiation therapy or chemotherapy may increase survival times -- Immunotherapy (Oncept canine melanoma vaccine by Merial) has recently become popular in dogs and seems to be safe for cats -- Believed to improve survival by inducing tumor-specific antibodies, cytotoxic T cells, and antitumor immune responses against melanomas Summary of cases - All Panthera were between 10 – 19 years old (4 female, 7 male) - classified as geriatric - 5/7 tigers were phenotypically white -- The majority of equine melanocytic neoplasms occur in grey or white horses with possible genetic cause -- Other explanation may be lack of protection to UV - Incisional biopsy diagnosed 9/11 cases - 9 animals had malignant melanoma only, 1 snow leopard only had a melanocytoma, and 1 tiger had both a melanoma and melanocytoma - Primary tumor sites included: -- skin of the eyelid, shoulder, brachium, pinna -- oral mucosa -- iris/uvea -- lip margin - The tumors were poorly pigmented in 3 tigers, 2 of white were white phenotype - Ocular tumors appeared as dark brown discolorations of the iris - 3/7 had pre-mortem diagnoses of metastasis to regional lymph nodes via FNA 7 cases underwent treatment - All received surgery, no correlation between completeness of excision and survival time -- Survival time ranged from 5 – 40 months (MST 8 months) - 4 received melanoma vaccines -- Survival time ranged from 8 – 40 months (MST 13.5 months) -- No adverse effects noted -4 underwent radiation therapy - 3/7 animals had regrowth of primary tumor after treatment - MST of treatment groups should be interpreted with caution due to mixture of euthanasia vs spontaneous death as well as small sample size Necropsy revealed metastasis of 7/9 animals with melanoma - Most common sites: lymph node (5/9), lung (4/9), liver (4/9), mesentery (4/9) - Mitotic activity was the best marker for histologically distinguishing malignancy - Majority of malignant melanomas had >4 mitotic activity per 10 high-power (X400) fields (1 malignant case had mitotic count of 1 per 10 hpf) - May need to stage every affected nondomestic felid regardless of mitotic activity Take-Home Message: - Both benign melanocytomas and malignant melanomas were seen in Panthera species including tigers, African lions, and snow leopards - Sites of tumors included skin, iris/uvea, oral mucosa, and lip margin - 5/7 tigers with malignant melanomas were the white phenotype - Treatments included combinations of surgery, radiation therapy, and melanoma vaccines - Melanoma vaccines deserve further study in treatment of melanocytic neoplasms in nondomestic felids

Answer 105

JZWM 2023 54(1):82-93 **DISEASE PROCESSES IDENTIFIED IN CAPTIVE ARABIAN SAND CATS (FELIS MARGARITA HARRISONI)** de Vargas AP, Jamorabon RE, Islam A, Kinne J ABSTRACT: The objective of this retrospective study is to identify common and significant causes of mortality and disease processes in the Arabian sand cat (Felis margarita harrisoni) captive population at Al Ain Zoo (Abu Dhabi, United Arab Emirates). Complete postmortem records of 25 Arabian sand cats, dead between 2009 and 2022, were reviewed retrospectively. A complete postmortem examination was done in all cases, and information was recorded in the Al Ain Zoo database and files. Out of 25 animals dead, 11 were adults (4-12 yr) and 12 were classified as geriatric animals (>12 yr), with only two neonatal (0-4 mon) deaths and no recorded deaths in juveniles (4 mon to 4 yr). Interestingly, but also expected because of the age range, 24% of the cases had concurrent pathologies at the time of death. **As expected in adult and geriatric felines, more than half of the cases (60%) developed nephropathies that were either one of the most important contributors or the main cause of death of the animal**. Different neoplastic lesions were described in four cases and reported for the first time in this subspecies: benign peripheral nerve sheath tumor, hepatobiliary carcinoma, and two different thyroid neoplasia. A vasculoproliferative disorder of the liver, peliosis hepatis, was described in one of the cases. **Additionally, in at least four cases, hyperthyroidism was strongly suspected in connection with thyroid neoplasia and hyperplasia, clinical signs, and other observed postmortem lesions**. Traumatic causes of death also were reported in six cases, including the only two neonates recorded dead. This information will contribute to Arabian sand cat improved veterinary care by identifying common pathologies in this species, potentially allowing earlier diagnosis and, ultimately, improving their management and husbandry in the captive breeding populations. Background: - Little information about Arabian sand cat pathology - Current conservation threats: -- Hybridization with domestic cats -- Transfer of known pathogens from domestic cats -- Both observed as a result of rapid urbanization Key Points: - 15/25 cats developed chronic renal insufficiency or related nephropathies -- Either the main cause or death or one of the main contributors -- In all cases, antemortem clinical signs as well as blood or urine markers were present -- Risk of CKD is likely due to genetic, age-related, environmental, and individual factors - 6/25 cats died of traumatic injuries -- Intraspecific aggression occurred most commonly in older male animals - 4/25 cat had suspected hyperthyroidism (but not confirmed antemortem via TT4 or fT4) -- All four had concurrent renal pathology; two cases had concurrent HCM - 4/25 cats had neoplasia -- Hepatobiliary and thyroid carcinoma; uterine leiomyoma; peripheral nerve sheath tumor - 3/25 cats died of Toxoplasma gondii -- All three were male adult animals -- Infection source unknown; suspect feral domestic cats - 1/25 died of Pasteurella multocida bronchopneumonia (considered an opportunistic infection) TLDR: CKD is common and often life-limiting in Arabian sand cats

Answer 106

JZWM 2023 54(1) 143-151 **UTILITY OF URINE ANTIGEN TESTING FOR DIAGNOSIS OF BLASTOMYCOSIS IN NONDOMESTIC FELIDS** Abstract: Blastomycosis caused by the fungus Blastomyces dermatitidis has been reported to cause disease in numerous species of nondomestic felids. Diagnosis of blastomycosis in domestic species often relies on the combination of clinical signs, radiographic findings, and commercial urinary antigen testing. **In this report, the sensitivity, specificity, and positive and negative predictive values for urine blastomyces antigen testing for use in nondomestic felids were examined and compared with findings on postmortem examination. The study showed a sensitivity of 100%, specificity of 91.86%, positive predictive value of 50%, and negative predictive value of 100% for urine antigen testing**. Furthermore, radiographic and hematologic findings were compared with those of animals diagnosed with blastomycosis. **Radiographic evidence consistent with blastomycosis was found in those animals diagnosed via urine antigen testing, but no significant differences in plasma biochemistry parameters between diseased and nondiseased animals were found. This study provides evidence that a positive blastomycosis antigenuria test result should be combined with other diagnostic methods to confirm the presence of infection with B. dermatitidis, whereas a negative antigenuria test result is 100% effective in predicting the absence of disease**. Intro - Blastomycosis is caused by the dimorphic fungus Blastomyces dermatitidis - Inhalation of fungal conidia can lead to pulmonic disease as well as the potential for systemic spread - direct inoculation via cutaneous routes can lead to infection, although this route is considered less common. - Generally not considered contagious or zoonotic - Currently, antemortem diagnosis of blastomycosis is based on cytology or histo - Culture not advised due to risk to lab personnel - In the absence of organism identification, diagnosis relies on a combination of clinical signs, radiographic findings, and commercial urinary antigen testing - Retrospective review of records of nondomestic felids tested for B. dermatitidis by using the urinary antigen test Objective: comparing these results to necropsy, radiographic imaging, and clinical findings to calculate the sensitivity, specificity, and positive and negative predictive values of the commercial urine antigen test. Results - Review of medical and necropsy records from 93 nondomestic felids yielded 7 animals considered true positive, 7 considered false positive, and 79 considered true negative - No animals were identified as false negative - Of these 93 animals, 60 (63.8%) were tigers (P. tigris) and 15 (16%) were lions (P. leo). Remaining were various other species - All animals considered true positive were tigers, six of the seven false positive animals were tigers, and one false positive was a lion - sensitivity of 100%, specificity of 91.86%, positive predictive value of 50%, and negative predictive value of 100% for urine antigen testing. - Of the seven true positive animals, five had detectable antigenuria within 7 d of postmortem examination. - Of the seven false positive animals, four had detectable antigenuria within 7 d of postmortem examination. - Radiographic evidence consistent with blastomycosis was found in the 5 true positives that had radiographs performed - Few differences in hematology parameters and no differences in biochemistry parameters between those animals identified as true positive versus false positive: clinical pathology alone should not be used as a means of diagnosing or justification for starting treatment for blastomycosis. -- In domestic species hypoalbuminemia is considered the most consistent abnormality, with mild hypercalcemia noted in approximately 10% of cases -- Of the 14 animals in this report in which blastomyces antigenuria was detected, no significant changes in bloodwork results were noted compared with standard reference values for these species - Possible cross-reaction of urine antigen test with histo (or other fungal dz) - Contact with the environment can lead to false positives–cystocentesis preferred **Takeaway–urine antigen blasto test in nondomestic felids has a low PPV but high NPV–a positive blastomycosis antigenuria test result should be combined with other diagnostic methods to confirm the presence of infection with B. dermatitidis, whereas a negative antigenuria test result is 100% effective in predicting the absence of disease**.