Sirenia Flashcards

Question

What is unique about the sirenian immune system? Which species have heterophils? Which have neutrophils? Basophils are common in which species? What is the most sensitive marker of inflammation?

Answer 1

Unique features * Has been postulated that they may have a **unique immune system whereby lymphocyte proliferation occurs as a first response to a stressor** * **However, when exposed to stressors such as brevetoxins this defensive mechanism is significantly suppressed—high likelihood of secondary infections** * Slight polychromasia in manatees and few nRBC in dugongs are considered normal * Rouleaux formation also considered normal * **Florida manatees and dugongs have heterophils rather than neutrophils** * Antilean manatees have neutrophils * Manatees can have a significant number of basophils; basophils are uncommon in dugongs * **Leukocyte count is a fairly insensitive marker of inflammation – total count may only reach 25K even with severe inflammation** * Other markers such as A:G ratio or SAA may have higher diagnostic value

Answer 2

Husbandry * Housing (FL manatees) * Freshwater or saltwater * If housed in saltwater, will thrive when provided freshwater for drinking * Water temperature: 25-30C (77-86F) * Filtration systems – remove large food and fecal bioloads * Chlorine (\<1 ppm) and ozone can be used, but excess oxidant use 🡪 eye damage * Should have a medical pool with false bottom floor or ability to drain rapidly * Social * Often respond well to presence of other manatees * Relieve anxiety * Encourage anorexic animals to eat * Not territorial or aggressive * Can be housed in groups (3-6 animals) with enough room * Be careful that males have not reached sexual maturity

Answer 3

* Diet * Obligate herbivores * Nutritional content differs between natural aquatic vegetation and commercial lettuce * Diets of mixed greens (lettuces, spinach, cabbage, kale), herbivorous primate pellets, variety of fruits, vegetables, grasses, hays * Manatee-specific pellet diet developed * Offer food at multiple levels in water column * Submerged feeders * Floating food * Partially submerged feeders along enclosure walls * FL manatees spend 4-8 hr/day eating * Adults consume 5-10% of BW daily * Food should be available 24 hr/day * Obesity is common

Answer 4

Handling and Restraint * Manatees can thrust paddles dorsoventrally or side-to-side * Place animals on thick closed-cell foam in sternal recumbency * Put large piece of closed-cell foam across the caudal body and paddle 🡪 have several people lean on foam for added restraint * Do not try to restrain an animal on its back * High risk of injury * Many are calm when removed from water * Most diagnostics and husbandry can be performed with mild restraint

Answer 5

Vascular access * Brachial vascular bundle--a rete of veins and arteries in the interosseus space between the radius and ulna * Can be accessed from the medial or lateral sides of the pectoral flipper * Caudal tail ventral vertebral arch vascular bundle is also used, especially in younger animals where the pectoral vessels are small * Not generally used in awake animals due to potential danger from movement of the fluke * Spinal needles can be used to obtain IV access for drug and fluid administration in anesthetized adults in the caudal tail vascular bundle * Blood collection * Brachial vascular bundle * Interosseous space b/w radius and ulna * Medial or lateral approach to pectoral flipper * Flipper firmly restrained 🡪 elbow and carpal joints identified 🡪 radius and ulna palpated 🡪 locate space between then 🡪 middle portion of flipper midway between elbow and carpus is surgically scrubbed * 18-20G, 1-2in needle attached to extension line and syringe or vacutainer * Calves * 25-21G butterfly set with ¾-in needle * Lateral aspect is easier due to minimal abduction of pectoral flipper * Blood clots quickly 🡪 heparinize needle * Caudal vascular bundle * Ventral to the vertebral column in the paddle * Difficult to access safely in awake animals * Use only in anesthetized or very ill animals

Answer 6

Endotracheal intubation · Often need to be mask induced prior to intubation, sometimes will breath hold which can make this difficult · Can theoretically be intubated orally or nasally, though oral is difficult due to poor visualization of the caudal pharynx and larynx due to curved angle of the tongue, narrow buccal cavity and long soft palate · Nasal intubation is best done with rhinoscopic assistance o Sometimes heavy loads of nasopharyngeal trematodes and subsequent rhinitis can complicate intubation · Intubation of a single bronchus can occur due to short trachea

Answer 7

Pre-anesthetic considerations · PE and bloodwork prior to anesthesia if possible · Ideally fast for 24 hours - though they have a slow gut transit time, so GIT will not be completely empty after 24 hours Induction/maintenance protocols · Despite low metabolic rate, similar drug doses tend to be effective as in other mammals · Midaz and diazepam are commonly used as premeds · Long needles must used to get through the skin and dermis · Combining detomidine and butorphanol has led to marked respiratory depression, though lower doses have been used without adverse side effects · Epidural with lidocaine can be useful for surgical procedures · Banamine can be used but only for a few days due to GI upset · Table in west of various drug combos that have been used · Mechanical ventilation during anesthesia is preferable · In some cases, whether due to disease or patient positioning, the peak inspiratory pressure of the ventilator must be adjusted above 20 mmHg to achieve lung expansion, oxygenation, and carbon dioxide exchange. Monitoring · Standard techniques Recovery considerations · Do not place back in water until they are appropriately responding to stimuli and breathing voluntarily · Preferable to place them in shallow water first so they can breathe by pushing off the pool floor with their pectoral flippers

Answer 8

* Watercraft injuries * Common cause of death of FL manatees and *T. manatus* in Puerto Rico * 22% of reported carcasses annually * Injuries on dorsum * Blunt trauma – boat hulls * Sharp/shearing trauma – propeller blades and lower motor units * Death from blunt trauma more common * Severe internal hemorrhage with minimal external wounds * Skin, diaphragm, and lung lacerations * Skeletal damage (rib or spinal fractures) * Hemothorax * Pyothorax * Pneumothorax * Hemoabdomen * Kidney damage * Pneumothorax * Clinical signs: increased RR and effort, listing, excessive buoyancy * Confirm with radiographs/ultrasound or air on thoracocentesis * Flotation devices may be necessary * Thoracocentesis * Diagnostic and therapeutic * Serial thoracocentesis can be successful for resolution * Remove smaller amounts over multiple procedures * If you remove to much, there will be immediate refilling of potential space * Chest tubes – challenging to maintain * Most resolve with supportive care and time * Thoracocentesis may not be indicated if animal is eating and breathing normally with no respiratory distress * **Boat strike is one of the most common causes of morbidity/mortality** * Death can result from sharp penetrating trauma from propellers and/or blunt trauma from the boat hull * Significant sequelae include hydrothorax, SQ emphysema, pneumothorax, secondary infections resulting in sepsis, pyothorax, pleuritis and peritonitis * Diaphragmatic hernias can also occur * Lungs often injured due to dorsal anatomic location * Fractures and luxations also occur **Watercraft injury** * One of the largest cases of rescue and mortality * Presence of external wounds in an animal with abnormal buoyancy is highly suggestive of traumatic pneumothorax * Excess intestinal gas from enterocolitis or constipation can also mimic these signs * Radiography to confirm * May need to tap repeatedly, severe cases may require the use of a flotation jacket to equalize the animal in the water column * Chest drains have had mixed results * Animals not responding to treatment after 4-6 weeks may have a severe tear in the lung tissue which may require surgical intervention * Head injury, spinal injury, and abdominal ST damage are also common sequela of boats trike * Propeller wounds are characteristic and can also cause rib transection * Proper wound care is key

Answer 9

* Entanglements * Very common reason for rescues of FL manatees Fishing gear * Ingestion of lines, hooks, lures, and netting material is common * Can cause perforations and death * Can be difficult to scope or remove surgically * Trapping in fish nets and drainage pipes can lead to drowning Other Environmental Health Concerns * Entrapment in floodgates, canal locks, drainage pipes, etc. * Fishhook foreign bodies * 2013 UME of FL manatees, bottlenose dolphins, brown pelicans in Indian River Lagoon * Dramatic reduction of seagrass due to long-term nontoxic phytoplankton blooms * Impacts of natural disasters and ecotourism being evaluated

Answer 10

**Cold Stress Syndrome (CRC)** * Prolonged exposure to temperatures \< 20C (68F) * Common in manatees, may also occur in dugongs * One of the major threats to free-ranging FL manatee populations * Sub-adults more commonly affected than adults * \> surface area:volume * Lack maternal care * Orphan calves susceptible due to lower fat mass * Acute exposure 🡪 lethargy and fatal hypothermia * Chronic exposure * Emaciation, depletion of fat stores, serous fat atrophy, lymphoid depletion, epidermal hyperplasia, pustular dermatitis, enterocolitis, myocardial degeneration * Opportunistic infections and secondary diseases * Bacteremia, dehydration, hypoglycemia, renal failure * Clin path * Thromboembolic disease * Hypercoagulability and have longer PT and PTT, increased D-dimer and fibrinogen, thrombocytopenia * Severe acidemia, metabolic acidosis, abnormal electrolyte findings * Leukocytosis * Elevated LDH, CK, creatinine, serum amyloid A * Clinical signs * Skin lesions and poor body condition * Early skin lesions: epidermal bleaching of extremities and muzzle * Chronic skin lesions: hyperkeratosis, diffuse pustules, ulcerative lesions * Muzzle, head, and extremities most severely affected * GI stasis, constipation, absence of gut sounds or flatulence, pneumonia **Cold stress syndrome (CSS) (ZP)** * A range of clinical and acute and chronic disease processes that can occur in manatees following exposure to water temps below 20 C (68 F) * Emaciation is noted in most chronic cases of CSS * Marked epidermal hyperplasia characterized by irregular, verruciform, pale grey plaques that are often fissured are commonly noted * Multifocal cutaneous ulceration and/or cutaneous pustules or abscesses * Histologically there is moderate to severe lymphoid depletion in LN, spleen, gut and respiratory mucosa * Myocardial degeneration * Heterophilic and histiocytic colitis or enteritis * Chronic-active suppurative bronchopneumonia * Pancreatic zymogen granule depletion * Acute skin lesions—vasculitis and vascular necrosis

Answer 11

* Brevetoxin (lipophilic, neurotoxic; PbTx) – bind to voltage-gated Na channels so that inactivation is prevented (cannot perform another action potential) * **Brevetoxicosis (CRC)** * West and east coasts of FL (west \> east) * Transmission by inhalation or ingestion * Toxins remain stable in environment and within seagrass * Mortalities seen long after or remotely located from known algal bloom * Clinical signs: neurologic and either respiratory or GI depending on route of transmission * Clin path abnormalities * Heterophilic and eosinophilic leukocytosis * Hemoconcentration * Increased PTT * Electrolyte abnormalities * Elevated CK * Hyperglobulinemia * Hypocalcemia * Decreased BUN, GGT, and amylase * Thrombocytopenia * Necropsy findings * Congestion of nasopharynx, airways, meninges * Hemorrhage in liver, kidney, lungs, +/- intestinal * Many found dead prior to intervention * Treatment * Prevention of drowning (flotation devices, propping on foam) * Supportive care (fluid therapy, anti-inflammatories, parenteral antibiotics, atropine * Animals with neuro signs generally recover within 24 hours * Other sequelae * Necrotic enteritis, pneumatosis intestinalis (PI), pyothorax, septic arthritis, sloughing of large portions of flippers * Treatment * Correct dehydration, acidosis, electrolyte abnormalities * If severe, IV fluids * Gastric tube with water 2-3 times/day over 24-48 hours 🡪 introduction of dilute gruel * Provide gruel or water until normal food consumption and fecal production seen * Offer food immediately and at all times * Constipation * Mineral oil and water via gastric tube, warm water or saline enemas * Broad-spectrum parenteral antibiotics * Secondary fungal infections have been seen * **Harmful algal blooms (HABs) - ZP** * Brevetoxicosis, caused by dinoflagellate **Karenia brevis** * Florida manatees often exposed to brevetoxins through consumption of filter feeding organisms or toxins directly associated * Exposure can also occur through inhalation of toxin, though there is no indication that inhalation alone can produce lethal doses * Respiratory lesions include catarrhal inflammation of the nasal sinuses and pulmonary edema * The neurotoxin produces nonsuppurative meningitis * Hemolytic toxins cause hemorrhage and hemosiderosis * Brevetoxins have an affinity for lymphocytes, macrophages, and microglial cells and their pathogenesis may involve elevated circulating cytokines and toxic shock * Diagnosis usually based on history, necropsy, and/or brevetoxin analysis

Answer 12

DNA viruses * **Trichechus manatus latirostris papillomaviruses (TmPV)** * Causes a range of hyperplastic skin lesions in florida manatees * The disease is largely historical and associated cutaneous lesions have not been described in free ranging animals * Papillomatous lesions, no inclusion bodies yet identified * Manifestation of the disease may involve viral latency and immunologic suppression

Answer 13

* Bacteria (CRC) * Dermal and SC abscesses not uncommon * Septic metritis secondary to dystocia and fetal maceration * Enteritis/colitis * Pneumonia * Pyelonephritis * Mycotic dermatitis * Pleuritis * Lung abscessation * Fatal atypical mycobacterial infections infrequently reported Bacteria (ZP) * Mycobacteriosis * M marinum, M fortuitum, M kansasii and M chelonae have been reported in captive sirenian * Multiple coalescing caseous nodules in the lung, pleura, LN, testicles, and skin * Salmonella * Can cause gastroenteritis in young sirenian * Salmonella lohbruegge has been described in a juvenile dugong causing granulomas in the liver * The respiratory tract is often involved in sirenian when bacterial infections occur * Antibodies to lepto and brucella have been reported, but no evidence of clinical disease

Answer 14

Fungi * Dermal mycosis * Has been reported in small numbers * Various fungal species isolated * May result in a cobblestone like appearance on the skin

Answer 15

* Parasites (CRC) * Rarely of clinical significance * *Cochleotrema cochleotrema* * Nasopharyngeal trematode 🡪 rhinitis * *Moniligerum blairi* and *Nudacotyle undicola*, small intestinal trematodes 🡪 enteritis in FL manatees * *Cryptosporidium* spp. 🡪 weight loss, diarrhea, abdominal discomfort, lethargy in Antillean and Amazonian manatees * *Toxoplasma gondii* uncommon * Encephalitis, myocarditis, disseminated toxoplasmosis reported * Exposure in Amazonian and West Indian manatees

Answer 16

Miscellaneous Conditions * Cardiac diseases rare * Cardiomyopathy, AV valve myxomatous transformation reported * Early reports suggests dugongs were susceptible to capture myopathy, but recent studies to not support this * Nephrolithiasis with pyelonephritis in 2 manatees * Polycystic kidneys in 1 FL manatee * Neoplasia infrequently reported * Reproductive neoplasia recently described in 8 FL manatees

Answer 17

* GI disorders * Constipation, diarrhea, dysbiosis, enterocolitis, colic-type clinical signs (rolling, abdominal crunching) * Periodic in adults, common in calves * Diagnostics: radiographs +/- contrast, fecal cytology, fecal occult blood, cultures for enteric pathogens * Supportive GI meds * Simethicone for colic-type signs * Metronidazole for clostridial enteritis/colitis * Transfaunation for dysbiosis * Equine probiotics * Constipation * Enemas * Metoclopramide * Gastric tube with mineral oil in water 2-3 times/day

Answer 18

* Analgesics * NSAIDS: flunixin meglumine, ketoprofen, aspirin, opioids (butorphanol and tramadol) * Oral tramadol at 1 mg/kg SID – reduced behaviors associated with pain (resting on bottom, crunching)

Answer 19

Poor buoyancy control * Support within the water column * Improves appetite, lowers stress during respirations, avoids chronic M/S complications, prevents drowning * Can use neoprene * Human flotation devices, foam added to the “down” side * Some animals will not tolerate them or eat with them on * Oral diazepam to help with acclimation to flotation device * If severely ill, keep in very shallow water to ease respiration * Should be propped on foam but remain afloat rather than dry-docked to avoid gravity-dependent injury * Attach food to side of the pool or at the bottom

Answer 20

Reproduction * FL and Amazonian manatees and dugongs = seasonal breeders * FL manatees * Estrous cycle = 28-42 days * Sexual maturity: 3-5 yrs, but may be related to size rather than age * Gestation: 14 months * Calves are dependent for ~2 years * Transplacental transfer of immunoglobulins documented in *T. manatus* * Single offspring, twinning is rare * Calving interval: 2.5-3yrs * Axillary teats * Nurse for 3-5 minutes every 1-2 hours * Polygynous * Herds of males pursue females 🡪 sometimes mortality of females from myositis and exhaustion * Dugongs * Sexual maturity: 10 yrs * Gestation: 13mo Reproduction (F8) * Females mature between 3-5 years of age * Dugongs mature at 10 years * Gestation 12-14 months in manatees and 13 months in dugongs * Calves nurse 1-2 years * Dystocia has been reported and sometimes requires C section

Answer 21

Neonatology and Hand-Rearing * Orphaned calves * Dehydrated, hypoglycemic, sometimes hypothermic * Initial assessment: blood glucose, PE, rectal temp * Initial treatment * Hypoglycemia and dehydration corrected with fluid and dextrose supplementation * Glucose \< 40 mg/dl require dextrose supplementation * Mild cases: gastric intubation with dextrose and oral electrolytes * Severe cases: IV administration * IV administration into a single vessel is difficult in a calf * Manatee immunoglobulin G via gastric tube or IV admin. * Within 24-48 hr 🡪 whole body radiographs and fecal sampling * Broad-spectrum antibiotics commonly used prophylactically * Nutritional support * Survival rates poor in orphaned calves \< 30 kg on admission * GI complications with artificial milk implicated as reason for poor success * GI disorders: diarrhea, constipation, decreased appetite, leukocytes in fecal cytology, fecal occult blood, necrotic enterocolitis * SeaWorld started using new milk formula recipes 🡪 successfully raised and released orphans with fewer GI complications * Transfaunation * Often helpful after recovery from PI or after treatment with oral antibiotics * From a healthy adult manatee 🡪 gastric intubation of a fecal slurry * Sometimes used in calves that have persistent diarrhea * First 3-4 weeks in rehab is the most critical period for orphans

Answer 22

Neonatal, maternally dependent calves * Orphans present to rehab facilities not uncommonly * Calves may suffer from complications related to prematurity, inanition, poor adaption, watercraft injury, and infection such as omphalitis, which includes hypoglycemia, dehydration, constipation, enterocolitis, and pneumatosis intestinalis. * Congenital defects in rescued calves have included ectrodactyly of the pectoral flippers digits, atresia ani, and omphalocele or exteriorization of the intestinal tract at the umbilicus * *Pneumatosis intestinalis* * GI gas within the wall of the GI tract * Secondary to necrotic enterocolitis, GI obstruction, GI ischemia, sepsis * Interactions with mucosal integrity, intraluminal pressure, bacterial flora, and intraluminal gas play roles in development * Treatment * Oral aminoglycosides * Metronidazole * Pepto-Bismol * Changing diet to elemental human infant formula

Answer 23

Physical Examination * RR: 3-5 breaths/ 5 mintues * Examine ventrum when dry-docked * Place straps under the axilla and cranial to the peduncle to facilitate rolling * Body condition * Malnourished * Distinct neck due to loss of nuchal fat (“peanut-head”) * Prominent scapulae, hips, spine * Flat (thin) or conave (very thin) ventrum * Longitudinal skin folds (emaciated) ![]() * Oral exam with digital palpation of molars * Thoracic and abdominal auscultation * Normal sinus rhythm * HR: 40-60 bpm * Calves: 60-75 bpm * Percussion useful for GI tract * Gas and peristaltic intestinal sounds are normal * Hindgut fermentation produces large volumes of colonic gas * Absence is abnormal Normal stool is formed, but not dry * If no feces, perform rectal exam

Answer 24

* Fecal sampling * Fresh feces 🡪 cytology, occult blood, parasitology * Collected an uncontaminated sample with a lubricated small flexible tube placed a short distance into the rectum * Gastric samples * Pass a standard soft plastic foal or small equine gastric tube through nares or oral cavity into stomach * Urethral catheterization * Difficult, not routinely performed * Urine samples * Place a small, flat, collection receptacle under the urogenital opening of a dry-docked manatee (a Frisbee!!) * Can stimulate urination - apply pressure on abdomen cranial to the vulva in females, caudal to genital opening in males

Answer 25

* Radiography and Ultrasonography * Radiographs * Fractures, pneumothorax, pneumonia, GI disorders * DV views are easier than VD * Orthogonal views are difficult if not impossible * Ultrasound * Pregnancy, subcutaneous abscesses, kidneys, urinary bladders, echocardiography * Guiding thoracocentesis for pneumothorax or pleural fluid accumulation * MRI/CT * For small animals * May or may not require sedation * Endoscopy * Flexible * Gastric, colonic, repro tract evaluation * Slow GI transit time limits GI tract visualization * Rigid thoracoscopy * Confirm pyothorax * Evaluate pleural and lung surfaces * Doppler for monitoring HR * Electrocardiography * Described in both species of *T. manatus* * Prolonged PR and QT intervals * Differences between adults and calves * Echocardiography parameters established

Answer 26

Clinical Pathology * Standard: CBC, chem, fibrinogen, ESR * When possible: D-dimers, serum amyloid A * Neonates, cold-stress, or critically ill: blood glucose * Manatee hematology * Larger erythrocytes * Lower erythrocyte counts * Heterophils rather than neutrophils * Stain positive for myeloperoxidase * Total WBC counts slightly lower than most * Differential: heterophils = lymphocytes * Differences b/w age classes, free-ranging vs captive, genders * Markers of inflammation * Total WBC count may only modestly increase with illness * Not sensitive indicator * Serum amyloid A – highly sensitive in FL manatees * Thromboelastography * Thromboembolic disease and DIC described in FL manatees * DIC most common in trauma and cold-stress syndrome * D Dimers, Fibronigen, PT, PTT

Answer 27

Therapeutics * IV fluid support * Calves and extremely ill individuals * Needle inserted into the brachial vascular plexus or ventral tail complex * Fluid support via gastric tube * More common * Oral or nasal * Inserted to the depth of the distal tip of the pectoral flipper * Start with conservative volume 2-3 times/day (more frequently in calves) * 1-2 L for adults * 30-100ml for calves * 0.5-1.5 L for sub-adults Nutritional support * Gruel: mixed lettuces, spinach, water, +/- herbivorous/omnivorous primate pelleted diet (monkey chow) * Gradual introduction of food * 25% strength at first, increased over several days once feces and flatulence are produced * Provide access to solid food at all times * Continue nutritional supplementation via gastric tube until eating normal amounts and producing normal stool for several days Oral medication * Administered most effectively via gastric tube * Only use oral antibiotics conservatively due to hindgut fermentation Parenteral medication * Parenteral antibiotics typically used * IM – caudal epaxial muscles * SC – shoulder or caudal epaxial areas * Shoulder muscles are very small * Surgical scrub should be used * Adults: 1.5-3.5in, 18-20G needle * 1.5-2 in. for SC, 3.5 in. for IM * Calves: 1 in. 20-22G needle * Duration of injection therapy should be limited when possible * Injection-site abscesses, pain, muscle necrosis due to chronic SC/IM injections

Answer 28

Barratclough, A., Ball, R. L., Floyd, R. F., Reep, R. L., & Conner, B. J. (2017). Proposal and application of a novel disseminated intravascular coagulation scoring system in the Florida manatee (Trichechus manatus latirostris). *Journal of Zoo and Wildlife Medicine*, *48*(1), 152-158. **Abstract:** Disseminated intravascular coagulopathy (DIC) is an acquired disorder of hemostasis resulting in activation of the coagulation and fibrinolytic pathways. It is reported secondarily to multiple disease processes and can be associated with increased mortality. Previous research at Tampa’s Lowry Park Zoo (LPZ) demonstrated that Florida manatees (Trichechus manatus latirostris) with cold stress syndrome (CSS) demonstrated thromboembolic disease. The object of this retrospective study was to establish the presence and clinical relevance of DIC in Florida manatees admitted to LPZ for rehabilitation from 07 March 2010 to 15 August 2015. A coagulation panel, including prothrombin time, partial thromboplastin time, platelet count, fibrinogen level, and D-dimer level was used to diagnose DIC. There were 100 cases identified in the study period: 35 trauma, 43 CSS, 17 secondary to harmful algae blooms (HAB), and five miscellaneous. Manatees with CSS had the highest incidence of DIC with 24 of 43 cases (56%) affected, followed by trauma with 18 of 35 cases (52%) affected. None of the manatees with HAB were found to have DIC. Manatees that developed DIC during rehabilitation or when DIC progressed did not survive. Due to the clinical implications of DIC, identifying its presence and recognizing its severity could improve clinical outcomes by enabling more intensive treatment protocols. * Disseminated intravascular coagulation (DIC): Acquired disorder of hemostasis * Always secondary to another cause (e.g. sepsis, trauma, neoplasia, immune-mediated disorders) * Diagnosis achieved through combination of tests * Coagulation panel: PT, PTT, D-dimers, fibrinogen, platelet count * Human ER medicine has validated a scoring system * Low fibrinogen is possible due to consumption * Developed a modified scoring system based on humans, cats, dogs. Combined score ≥ 4 is associated with DIC * Cold stress syndrome (CSS) and trauma are likely associated with DIC (56% and 52% of cases, respectively) * DIC was not found in harmful algal bloom (brevetoxicosis) cases * Trauma had the highest mortality rates compared to CSS and HAB cases. Reduced survival in trauma manatees with DIC. * In CSS, DIC is common (56%) though survival probability is not related to DIC presence or absence * Can correct underlying problem with supportive care and resolution of hypothermia * If repeat coag panels, a change in DIC score was a better predictor of outcome * D dimers were best standalone indicator for long-term prognosis * \> 1,500 ng/mL = poor w/ only one survivor (3200 ng/mL) **Summary:** Over half of cold-stressed manatees had DIC but it was not associated with poorer long-term outcome. Over half of trauma cases had DIC and it was associated with reduced survival. There was no DIC in brevetoxicosis cases. D-dimers were the best single test for predicting outcome.

Answer 29

Martony, M. E., Isaza, R., Erlacher-Reid, C. D., Peterson, J., & Stacy, N. I. (2020). Esophageal measurement of core body temperature in the florida manatee (trichechus manatus latirostris). *Journal of Wildlife Diseases*, *56*(1), 27-33. ABSTRACT: Cold-stress syndrome (CSS) is one of the leading natural threats to free-ranging Florida manatees (Trichechus manatus latirostris). Cold water exposure below the species’ acceptable physiologic range is a frequent occurrence for manatees during cold weather months causing CSS induced systemic illness and significant annual mortality. Although CSS is a commonly presented condition at manatee rehabilitation facilities, the core body temperatures in CSS manatees are currently unknown due to the lack of clinically applicable and accurate temperature measurement methodologies**. Our objective was to establish a clinically applicable measurement methodology of core body temperature in manatees.** A novel, minimally invasive temperature technique to obtain esophageal temperature by placing a temperature sensor through an oro-gastric tube was compared to current oral and nasal methods in 20 clinically healthy manatees. **Results identified the esophageal measurement as the best performing and most precise temperature methodology.** The superior performance of esophageal temperature measurements differed significantly from both nasal and oral measurements, while nasal and oral measurements did not differ when compared with each other. **The esophageal measurements were consistent with manatee core body temperature, facilitating generation of a reference interval for core body temperature in healthy manatees (35.0– 35.8 C).** Four CSS medical cases were evaluated with the newly validated esophageal temperature method, facilitating diagnosis of hypothermia. The application of this temperature measurement technique to CSS manatees in field or rehabilitation settings will help in understanding CSS pathophysiology, improve medical assessments during rehabilitation, and contribute to conservation efforts. Intro * Cold stress syndrome (CSS) is one of the leading threats to wild Florida manatees * No accurate method for assessing core body temp in manatees exist * objective was to establish a clinically applicable measurement methodology of core body temperature in manatees M&M * n = 20 healthy manatees * **Measurement of esophageal temps via orogastric tube with a digisense thermometer. This was compared to oral and nasal measurements** * Temperatures from 4 manatees that were clinical for CSS were also measured via esophageal method and were compared to the healthy manatee measurements Results and discussion * The esophageal measurement was significantly different (higher) from oral and nasal * Also consistent with core body temperatures measured in research settings with more invasive techniques * Oral and nasal were not significantly different from each other * The reference interval for core body temperature as measured by esophageal temperature for the healthy manatees was 35.0–38.5 C (95-100.4 F) * The esophageal temperatures of all 4 CSS manatees were below this reference range by at least 1.2 deg C * The procedure was well tolerated by all manatees and temperatures were attained quickly and remained stable over a 2-minute period * Note that measurements were taken after 5 minutes out of water, so could be some environmental impact (warmer air compared to the water leading to increase core temp) on temperature though none was identified **Takeaway:** Esophageal temperature measurements is an accurate and non-invasive way to measure core body temperature in florida manatees

Answer 30

Journal of Zoo and Wildlife Medicine 48(1): 13–17, 2017 **IDENTIFICATION OF BACTERIA IN BLOOD CULTURES FROM CLINICALLY ILL CAPTIVE ANTILLEAN MANATEES (TRICHECHUS MANATUS MANATUS)** Mariana C. O. Silva, D.V.M., M.Sc., Fernanda F. L. Attademo, D.V.M., Ph.D., Augusto C. B. Freire, D.V.M., M.Sc., Glaucia P. Sousa, D.V.M., Fábia O. Luna, B.Sc., Ph.D., Débora C. V. Lima, D.V.M., M.Sc., Rinaldo A. Mota, D.V.M., Ph.D., Emiko S. Mendes, D.V.M., Ph.D., and Jean C. R. Silva, D.V.M., Ph.D. **Abstract:** Between September 2001 and March 2013, 62 bacterial cultures (37 aerobic and 25 anaerobic) were performed on 37 blood samples from 23 Antillean manatees (Trichechus manatus manatus) that were kept in captivity at the Brazilian National Center for Research and Conservation of Aquatic Mammals (CMA) in Pernambuco (CMA-PE) and Alagoas (CMA-AL), Brazil. All of the animals sampled exhibited clinical signs at the time of sampling including abscesses (n ¼ 8), debilitation and anorexia (n ¼ 22), and profound lethargy moribundity (n ¼ 7). The 4 animals with profound lethargy-moribundity died shortly after sampling of unknown causes. Bacteria were isolated from 15/37 (40.5%) and aerobic blood cultures from 13/23 animals (56.5%). None of the anaerobic cultures were positive. Aeromonas caviae, Aeromonas hydrophila, Aeromonas sp., Escherichia coli, Leclercia adecarboxylata, Pantoea agglomerans, Pseudomonas aeruginosa, Pseudomonas stutzeri, Pseudomonas sp., Sphingomonas paucimobilis, coagulase-negative Staphylococcus, and Staphylococcus epidermidis were each found in only one animal; Staphylococcus spp. was found in two; and Vibrio fluvialis in four. Thirteen samples had only one bacteria isolated, one sample had two bacteria, and one sample had three bacteria isolated. **Regarding sex, age group, and origin among the manatees examined, 54.5% (6/11) of the females, 58.3% (7/12) of the males, 40% (2/ 5) of the calves, 66.7% (8/12) of the juveniles, 50% (3/6) of the adults, 55.5% (10/18) at CMA-PE, and 60% (3/5) at CMA-AL were found to be positive for bacterial growth during at least one sampling time.** All Antillean manatees were clinically ill. **Regarding clinical signs, bacteria were found in 50% (11/22) of blood samples of the animals showing debilitation and anorexia, 1 of 8 (12.5%) of blood samples of the animals showing abscesses, and 3 of 7 (42.9%) of blood samples of the animals showing profound lethargy-moribundity**. * Antillean manatee – IUCN critically endangered * Debilitated manatees – blood culture * Staph – different virulence * Pseudomonas & vibrio – common in aquatic environments * Aeromonas – common in aquatic environments – produces opportunistic disease, pneumonia in dugongs * E. coli isolated in a few **Take Home:** Debilitated manatees are often septic – consider blood culture if one presents to you.

Answer 31

Ferrante, J. A., Hunter, M. E., & Wellehan, J. F. (2018). Development and validation of quantitative PCR assays to measure cytokine transcript levels in the Florida manatee (Trichechus manatus latirostris). *Journal of wildlife diseases*, *54*(2), 283-294. ABSTRACT: Cytokines have important roles in the mammalian response to viral and bacterial infections, trauma, and wound healing. **Because of early cytokine production after physiologic stresses, the regulation of messenger RNA (mRNA) transcripts can be used to assess immunologic responses before changes in protein production**. To detect and assess early immune changes in endangered Florida manatees (Trichechus manatus latirostris), **we developed and validated a panel of quantitative PCR assays to measure mRNA transcription levels for the cytokines interferon (IFN)-c; interleukin (IL)-2, -6, and -10; tumor necrosis factor-a; and the housekeeping genes glyceraldehyde3-phosphate dehydrogenase (GAPDH) and b-actin (reference genes).** Assays were successfully validated using blood samples from free-ranging, apparently healthy manatees from the east and west coasts of central Florida, US. **No cytokine or housekeeping gene transcription levels were significantly different among age classes or sexes. However, the transcription levels for GAPDH, IL-2, IL-6, and IFN-c were significantly higher (P,0.05) in manatees from the east coast of Florida than they were from those from the west coast.** We found IL-10 and b-actin to be consistent between sites and identified b-actin as a good candidate for use as a reference gene in future studies. Our assays can aid in the investigation of manatee immune response to physical trauma and novel or ongoing environmental stressors. * Our purpose was to optimize and validate a panel of real-time qPCR assays targeting cytokines associated with the immune response in manatees. * Collected blood from 30 free ranging Florida manatees during routine health assessments * Developed and validated a panel of quantitative PCR assays to measure mRNA transcription levels for the cytokines interferon (IFN)-c; interleukin (IL)-2, -6, and -10; tumor necrosis factor-a; and the housekeeping genes glyceraldehyde3-phosphate dehydrogenase (GAPDH) and b-actin (reference genes) * No cytokine or housekeeping gene transcription levels were significantly different among age classes or sexes. However, the transcription levels for GAPDH, IL-2, IL-6, and IFN-c were significantly higher (P,0.05) in manatees from the east coast of Florida than they were from those from the west coast. * The b-actin gene showed promise as a reference gene for manatee studies because it did not differ between sex, age class, or location. However, because of the observed difference in GAPDH expression between sites, its use as a reference gene may need to be restricted to site-specific studies. West (Gulf Coast) has more environmental pollutants (worse blooms and effects of oil)

Answer 32

Borges, J. C. G., Jung, L. M., Santos, S. S. D., Carvalho, V. L., Ramos, R. A. N., & Alves, L. C. (2017). Treatment of Pulmonicola cochleotrema infection with ivermectin-praziquantel combination in an antillean manatee (Trichechus manatus manatus). *Journal of Zoo and Wildlife Medicine*, *48*(1), 217-219. Abstract: The aim of this study was to report the use of an oral combination of ivermectin plus praziquantel in the treatment of a **Pulmonicola cochleotrema** in an Antillean manatee (Trichechus manatus manatus). A female manatee was found exhibiting **respiratory changes** **and the** **presence of parasites in the nares**. Based on clinical manifestations presented by the manatee, a symptomatic therapeutic protocol was employed, which included an anthelmintic treatment using a combination of **ivermectin plus praziquantel**. The parasites retrieved were identified as P. cochleotrema. The **fourth day** after the onset of the therapeutic protocol, the **clinical signs declined** and on the **seventh day posttreatment no clinical** **signs were observed**. This is the first time a therapeutic protocol of ivermectin plus praziquantel has been used in the treatment of P. cochleotrema in manatees. * All the fecal samples analyzed (before and after treatment) did not show the presence of helminths eggs * Asymptomatic or respiratory signs associated with rhinitis, pulmonary edema, pneumonia **Take** **home**: Successful treatment of Pulmonicola cochleotrema using ivermectin and praziquantel in manatee

Answer 33

EVALUATION OF THE PHARMACOKINETIC BEHAVIOR OF TULATHROMYCIN (DRAXXIN) IN FLORIDA MANATEES (TRICHECHUS MANATUS LATIROSTRIS) UNDERGOING MEDICAL REHABILITATION. Smith LN, Bublitz C, Nixon E, Yeatts J, Ball RL, Baynes RE. Journal of Zoo and Wildlife Medicine. 2021;52(3):880-885. Florida manatees (Trichechus manatus latirostris) frequently present to rehabilitation care facilities for various conditions, including boat strike trauma, cold stress syndrome, and brevetoxicosis. Throughout the course of treatment, antimicrobial use to treat respiratory disease is frequently warranted. To date, clinicians have extrapolated dosages based on established information available in bovine and equine medicine. The routes of administration, efficacy, and treatment intervals are considerations in dealing with critical wild animals. The use of tulathromycin, a triamilide antibiotic, has been studied in multiple domestic species of economic importance, including cattle, small ruminants, and swine, and has revealed efficacy against respiratory diseases. Given this information, this antibiotic has also been used in manatees with positive clinical outcomes. This study employed **sparse sampling and evaluated banked plasma samples at various time intervals post–tulathromycin administration obtained during the clinical treatment course of nine animals during their rehabilitation. Preliminary pharmacokinetic analysis following administration of a single dose** estimated a half-life of 33.75 h and volume of distribution per fraction absorbed (Vz/F = 4.29 L/kg). The pharmacokinetic behavior of tulathromycin in Florida manatees can be used to optimize dosage regimens in this species. **Background** * FL manatee down-listed from endangered to threatened in 2017; no PK studies in manatees * Tulathromycin: macrolide, high concentrations in lungs compared to plasma in pigs, mice, horse, and cattle * Accumulates in neutrophils and alveolar macs * Current tulathromycin dose routinely used in manatees: 2.5 mg/kg SQ q7d * Respiratory isolates in a study of Antillean manatees: G- (*E coli, Enterobacter sakasakii, Providencia rettgeri, Stenotrophomonas maltophilia*) **Key Points** * 1-3 retrospective samples from each time point, some different and some the same animals, animals were sick (brevetoxin, entanglement, boat strike, cold stress, orphaned calf) * Single dose 2.5 mg/kg SQ: rapid elimination and short half life (34 hr), fell below 0.1 ug/ml by 120h * Shorter than cattle (44-90h depending on age and route of admin) * Two doses 9 days apart: prolonged elimination phase and long half life (280 hr), fell below 0.1 ug/ml after 216h * Large volume of distribution supports hypothesis that despite dropping plasma levels, may concentrate in tissues, especially lungs * Unknown if tulathromycin accumulates in heterophils of manatees like it does in neutrophils of cattle **Conclusions** * Tulathromycin can reach clinically applicable PK values in the manatee (above 0.03 ug/ml for 5 days) * Multidose regimen (2.5 mg/kg q7-9d) is supported based on extended half life and plasma conc (9days)

Answer 34

Baseline urinalysis results in 32 healthy Antillean manatees (Trichechus manatus manatus) JAVMA 2021;258:416–424 OBJECTIVE **To describe results of analysis of free-catch urine samples collected from Antillean manatees (Trichechus manatus manatus) under human care in the Caribbean**. ANIMALS **32 Antillean manatees** in 5 Caribbean oceanaria and rescue centers. PROCEDURES Urine samples were obtained by opportunistic free catch during physical examination or through the use of operant conditioning procedures. Urinalyses consisted of macro- and microscopic evaluations, biochemical analyses with test strips, and refractometry. Results were compared for manatees grouped on the basis of age, sex, and habitat. RESULTS **Urine samples were typically clear, straw colored, and alkaline (mean pH, 8.0); had a urinoid odor and low specific gravity (mean, 1.010); and had results on qualitative test strips that were consistently negative for the presence of glucose, bilirubin, ketones, proteins, nitrites, RBCs, and WBCs.** Microscopically, the mean ± SD number of RBCs and WBCs/hpf was 0.5 ± 0.3 RBCs/hpf and 1.1 ± 1.5 WBCs/hpf. **The presence of some epithelial cells and crystals was typical. Spermatozoa were found in urine from 1 of 15 sexually mature males, and parasite larvae and eggs were found in urine from 2 manatees**. CONCLUSIONS AND CLINICAL RELEVANCE Results of the present study yielded the first compilation of baseline urinalysis values in healthy Antillean manatees under human care, which, when combined with physical examination and other diagnostic procedures, can help in monitoring the health of these animals. We encourage the use of free-catch urine collection methods, as used in the present study, for routine urinalyses of manatees under human care in zoos, aquaria, or rescue centers. Intro * Antillean manatees (Trichechus manatus manatus) are endangered,a euryhaline, herbivorous aquatic mammals that live in marine, estuarine, and freshwater ecosystems of the Caribbean * Our aim of the study reported here was to describe the macroscopic, microscopic, and biochemical characteristics of free-catch urine samples collected through the use of noninvasive medical operant conditioning in Antillean manatees under human care in the Caribbean. M&M * Thirty-two Antillean manatees (21 males (15 sexually mature), 11 females(8 sexually mature)) under human care at 5 oceanaria and rescue center facilities * Free catch UA * Gross examination, biochemical analysis and microscopic evaluation Results and discussion * The color of most urine samples was straw (22/32 [69%]), followed by pale yellow (6/32 [19%]) or colorless (4/32 [13%] * Consistent with other herbivorous species * the appearance of most samples was clear (21/32 [66%]), and the most common odor of samples was urinoid (strong urine smell; 25/32 [78%] * qualitative test strips that were consistently negative for the presence of glucose, bilirubin, ketones, proteins, nitrites, RBCs, and WBC * In contrast, Amazonian manatees have been reported8 to have various numbers of WBCs and RBCs and even suppurative exudate in their urine, suggesting the presence of genitourinary infections. * semiquantitative trace concentrations were detected for urobilinogen (0.2 mg/dL; n = 32/32), creatinine (0.1 mg/dL; 17/17), and microalbumin (10 mg/dL; 17/17 * the mean ± SD specific gravity measured by refractometry was 1.010 ± 0.007 and did not differ substantially between males (1.011 ± 0.008) and females (1.009 ± 0.005) * Our findings suggested that the capacity to concentrate urine is similar between Antillean manatees and Amazonian manatees * the mean ± SD urine pH was 8.0 ± 0.9 and did not differ substantially between males (7.9 ± 1.1) and females (8.0 ± 0.7) * **Marine mammals, such as common bottlenose dolphins, gray seals (Halichoerus grypus), and South American sea lions (Otaria byronia), tend to have acidic urine, and a previous study shows that Florida manatees also have acidic urine** * Qualitatively, **epithelial cells** were observed in urinary samples from 28 manatees, with transitional cells less commonly present (1/32 [3%]), compared with **squamous cells (**21/32 [66%]), which were more commonly identified in samples from females (11/11 [100%]) versus males (10/21 [48%]; * Urinary crystals were observed in samples from 13 manatees, with **calcium oxalate monohydrate crystals** most commonly identified among the affected samples (n = 4) * There were rare observations of mucus, erythrocyte casts, hyphae, sperm, or parasite larvae and eggs, and these were found only in samples from adult male manatees. Take home: free catch UA parameters established in healthy Antillean manatees

Answer 35

PRESENTATION AND MANAGEMENT OF DENTAL PAD FRACTURES IN TWO FLORIDA MANATEES (TRICHECHUS MANATUS LATIROSTRIS) Journal of Zoo and Wildlife Medicine 52(3): 1084–1089, 2021 Abstract: **The Florida manatee (Trichechus manatus latirostris) has well-developed keratinized dental pads at the most rostral aspect of their mouth to assist with mastication.** This unique development is thought to be an adaptive response to their highly abrasive diets that contain phytoliths and sediments that may accelerate dental wear**. In May 2013, two Florida manatees presented with multiple fractures in their inferior dental pads.** **The fractures were successfully managed with nutritional modifications, dental pad trimming, and vigilant monitoring through behavioral husbandry training.** Signs of spontaneous healing were observed as early as 60 days after initial presentation with subsequent full resolution. Although surgical intervention was planned, the spontaneous healing mitigated significant health risks associated with the procedure. **To the authors’ knowledge, these are the first reported cases of dental pad fractures and their spontaneous healing and resolution in manatees.** Intro * Manatees have well-developed keratinized dental pads at the front of their mandibles, rostral to the cheek-teeth * These heavily cornified pads provide a hard surface area to mechanically break down sea grasses during mastication and before transit to the cheek-teeth Animal 1 * 33 yo MI florida manatee * Presented with 2 fractures of the inferior dental pad during a routine session * Weekly trimming under behavioral restraint was implemented * Appeared visually healed at day 60 aside from a hairline fracture mark * Fully resolved 3 months later Animal 2 * 36 yo MI florida manatee * Lived in managed care, cohabitates with animal 1 * Inferior dental pad fracture observed during routine training session (2 weeks after animal 1) * By day 60 the pad was visually normal * No further fractures for more than 5 years Treatment and management * Food items were tailored to minimize pressure points during mastication (ie lettuce offered as leaves only with the based removed, monkey biscuits soaked etc) * Close examination of enrichment items was performed to ensure that the manatees couldn’t bite them and cause further damage * Because Animal 1 had fractures that were more severe and chronic, the loose part of the fracture did not wear down on its own and thus required dental pad trimming on a weekly basis to prevent a fulcrum * Although surgery to remove the loose tissue on Animal 1 was planned, both initial fractures spontaneously healed after 60 days, making surgical intervention no longer necessary Discussion * this case study demonstrates that dental pads can spontaneously heal without the need for surgical intervention. * It is unknown if there are genetic or nutritional risk factors involved or what the frequency of this clinical presentation may be in the wild, but the authors hypothesize that this may be underdiagnosed, and it warrants further exploration. * A possible etiology for the dental pad fractures is that the manatees may have been cribbing more frequently because of discomfort from an underlying tooth issue. * Cribbing was observed from both animals, which sparked the initial examination, but no other clinical or behavioral concerns were identified. * Because both animals presented with the condition very close in time, another possible etiology is that new enrichment items or some other husbandry issue were introduced and caused trauma to the dental pads.