Splenic marginal zone lymphoma in an Asian small-clawed otter (Aonyx cinerea).

Severe splenomegaly was found during routine examination of a clinically normal 7-yr-old male Asian small clawed otter. The spleen and three enlarged splenic lymph nodes were immediately removed. The spleen weighed 310 g (approximately 8% of body weight). The spleen and resected lymph nodes were diffusely infiltrated by coalescing sheets of neoplastic lymphocytes that occasionally surrounded remnants of preexisting lymphoid follicles. Immunohistochemical confirmation of B lymphocyte origin and microscopic pattern were consistent with primary splenic marginal zone lymphoma (MZL) with metastasis to the splenic lymph nodes. The otter received no additional treatment and survived for 16 mo following splenectomy. Necropsy confirmed metastasis to multiple abdominal and extra-abdominal lymph nodes, liver, and kidney, and renal failure related to glomerulosclerosis. The prolonged survival in this otter is typical for MZL, an indolent form of B-cell lymphosarcoma that spreads slowly to the abdominal and extra-abdominal lymph nodes.

Cloacolithiasis and intestinal lymphosarcoma in an African black-footed penguin (Spheniscus demersus).

A 13-yr-old male African black-footed penguin (Spheniscus demersus) presented thrice over 7 mo with gastrointestinal obstruction secondary to cloacolithiasis. Clinical signs consistently resolved with cloacolith removal and supportive care. However, 10 mo after initial presentation, it presented with similar signs, plus significant weight loss. No cloacolith was found, and it subsequently died. Significant gross findings included bilateral cecal masses, colonic perforation, and marked secondary coelomitis, multifocal tan to pale hepatic nodules, and pale kidneys with miliary white foci. Histopathologic diagnoses were intestinal lymphosarcoma with hepatic and renal metastases, secondary intestinal rupture, and subacute severe bacterial coelomitis. To the authors' knowledge, this is the first full report of either cloacolithiasis or lymphosarcoma in a penguin.

Cytosolic phospholipase A2α blockade abrogates disease during the tissue-damage effector phase of experimental autoimmune encephalomyelitis by its action on APCs.

Cytosolic phospholipase A(2)α (cPLA(2)α) is the rate-limiting enzyme for release of arachidonic acid, which is converted primarily to PGs via the cyclooxygenase 1 and 2 pathways and to leukotrienes via the 5-lipoxygenase pathway. We used adoptive transfer and relapsing-remitting forms of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, in two different strains of mice (SJL or C57BL/6) to demonstrate that blockade of cPLA(2)α with a highly specific small-molecule inhibitor during the tissue-damage effector phase abrogates the clinical manifestation of disease. Using the adoptive transfer model in SJL mice, we demonstrated that the blockade of cPLA(2)α during the effector phase of disease was more efficacious in ameliorating the disease pathogenesis than the blockade of each of the downstream enzymes, cyclooxygenase-1/2 and 5-lipooxygenase. Similarly, blockade of cPLA(2)α was highly efficacious in ameliorating disease pathogenesis during the effector phase of EAE in the adoptive transfer model of EAE in C57BL/6 mice. Investigation of the mechanism of action indicates that cPLA(2)α inhibitors act on APCs to diminish their ability to induce Ag-specific effector T cell proliferation and proinflammatory cytokine production. Furthermore, cPLA(2)α inhibitors may prevent activation of CNS-resident microglia and may increase oligodendrocyte survival. Finally, in a relapsing-remitting model of EAE in SJL mice, therapeutic administration of a cPLA(2)α inhibitor, starting from the peak of disease or during remission, completely protected the mice from subsequent relapses.

Cutaneous and splenic mastocytosis in a juvenile Malayan tiger.

A male Malayan tiger cub developed well-circumscribed, erythematous, alopecic lesions on the face, torso, and paws when 1-wk-old. Biopsies of a torso lesion and a right front paw lesion at 1-mo-old confirmed cutaneous mast cell tumors (MCTs). MCTs on the paws grew into pendulous masses up to 6.5 cm in diameter by 3-mo-old, but those on the face and torso regressed. Fine-needle aspiration of the spleen at 3-mo-old revealed marked mast cell infiltration. The spleen and the right paw cutaneous MCT were removed; the paw MCT recurred within 7 d. A 12-bp tandem duplication, suggesting a somatic mutation, was identified in exon 8 of c-KIT in DNA extracted from the cutaneous MCT on the right paw and from one over the torso, but not from the spleen. Remaining MCTs on the paws regressed slowly following splenectomy and had completely regressed by 1-y-old. At 7-y-old, there was no recurrence of any mast cell disease. Mast cell disease in this tiger cub is similar to a report in a domestic kitten and to pediatric mastocytosis in humans, which commonly begins in infancy, improves by adolescence, and is associated with somatic c-kit mutations. To our knowledge, mastocytosis has not been reported previously in a juvenile exotic felid.

Pharmacokinetics of meloxicam following intravenous and oral administration in green iguanas (Iguana iguana).

OBJECTIVE: To determine pharmacokinetics of meloxicam in healthy green iguanas following PO and IV administration and assess potential toxicity. ANIMALS: 21 healthy green iguanas (Iguana iguana). PROCEDURES: To assess pharmacokinetics, 13 iguanas were administered a single dose (0.2 mg/kg) of meloxicam PO and, 14 days later, the same dose IV. To assess potential toxicity, 4 iguanas were given meloxicam at a dosage of 1 or 5 mg/kg, PO, every 24 hours for 12 days, and results of histologic examination were compared with results for another 4 iguanas given a single dose of meloxicam (0.2 mg/kg). RESULTS: There were no significant differences between PO and IV administration with regard to terminal half-life (mean ± SD, 12.96 ± 8.05 hours and 9.93 ± 4.92 hours, respectively), mean area under the curve to the last measured concentration (5.08 ± 1.62 µg•h/mL and 5.83 ± 2.49 µg•h/mL), volume of distribution (745 ± 475 mL/kg and 487 ± 266 mL/kg), or clearance (40.17 ± 10.35 mL/kg/h and 37.17 ± 16.08 mL/kg/h). Maximum plasma concentration was significantly greater following IV (0.63 ± 0.17 µg/mL) versus PO (0.19 ± 0.07 µg/mL) administration. Time from administration to maximum plasma concentration and mean residence time were significantly longer following PO versus IV administration. Daily administration of high doses (1 or 5 mg/kg) for 12 days did not induce any histologic changes in gastric, hepatic, or renal tissues. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that administration of meloxicam at a dose of 0.2 mg/kg IV or PO in green iguanas would result in plasma concentrations > 0.1 µg/mL for approximately 24 hours.

Diagnosis and Surgical Management of a Paratesticular Cyst in a Rehabilitating Juvenile Male Green Turtle (Chelonia mydas).

A juvenile green turtle (Chelonia mydas) undergoing rehabilitation for cold stunning exhibited an asymmetric bulging of the left caudal plastron and was diagnosed with a large intra-coelomic mass based on radiographical findings. Ultrasonography further identified a fluid-filled structure within the caudal coelom. Cytological evaluation of fluid obtained from the structure was consistent with a transudate, and thus, a cyst of unknown origin was suspected. Computed tomography imaging was pursued to further characterize the extent and location of the mass, which occupied ~50% of the total coelomic cavity volume. Conservative management with monitoring and occasional drainage of the mass did not result in improvements; thus, an exploratory laparoscopy for further investigation and surgical planning was elected. Intra-coelomic surgery was performed to remove a thick-walled cystic mass associated with the left gonad. Histopathology confirmed a paratesticular cyst continuous with, and possibly originating from, the epididymis. Post-surgical recurrence of the cyst was not appreciated, and the animal was successfully released 1 year after admission. Unrelated to the cyst, the turtle developed acute severe anemia on two occasions throughout rehabilitation that responded to modification of antimicrobial treatment and subsequent steroid administration. To the authors' knowledge, this is the first report of a paratesticular cyst in a reptile.

Epidemiological Investigation of a Mortality Event in a Translocated Gopher Tortoise (Gopherus polyphemus) Population in Northwest Florida.

Nokuse Plantation, a 22,055 ha private conservation preserve in northwest Florida, is a recipient site for gopher tortoises translocated from development sites in Florida. Since 2006, Nokuse has received over 5,000 tortoises from multiple development sites. During 2013-2015, 52 tortoises were found sick (n = 14) or dead (n = 38) in multiple soft-release enclosures in which tortoises consistently exhibited clinical signs, with additional sick (n = 5) and dead (n = 5) tortoises presenting similarly during 2016-2017. When found alive, tortoises behaved abnormally (e.g., frequently out of burrows during cold weather, pacing along enclosure fencing), appeared emaciated, were lethargic, and had developed redness under plastron scutes. Similar numbers of male (n = 28) and female (n = 32) tortoises were recovered along with two of unidentified sex, including mainly adults (n = 59) and three subadults. Physical examination, blood analysis, and other diagnostics were indicative of starvation and dehydration. Most sick tortoises provided with supportive care recovered. Necropsy findings generally confirmed starvation, with no evidence of infectious pathogens or contaminants. There were no apparent differences in quality of habitat, plant community, or soil or water among affected and unaffected enclosures. Botanical surveys indicated adequate forage quality and quantity, with no poisonous exotic or native plants detected. No land management practices changed prior to this event. Analysis of epidemiological data and demographic factors from before and during this mortality event identified initial density of tortoises in the enclosures as exerting the strongest influence on detection of tortoise morbidity and mortality. We believe that the stress associated with mixing tortoises from different populations and at higher densities during translocation impacted an individual tortoise's ability to obtain or absorb adequate nutrients from foraging, ultimately leading to a wasting condition consistent with starvation. Based on our findings, we recommend a maximum of 3 gopher tortoises per ha in soft-release enclosures for translocation, but further research is warranted to investigate the complexity of stress and social pressures associated with translocation.

Proventricular nematodiasis in wrinkled hornbills (Aceros corrugatus).

Three immature Sunda wrinkled hornbills (Aceros corrugatus) were diagnosed postmortem with proventricular spirurid nematodiasis. Concurrent severe disseminated larval granulomatosis in other visceral organs was considered contributory to mortality in each case. Clinical signs of nematodiasis were vague but generally consisted of weight loss, anorexia, and lethargy. Frequent antemortem fecal examinations were negative for spirurid eggs. In these present cases, based on routine histopathology, both prophylactic and empirically based therapeutic anthelmintic treatments had no evident benefit in the elimination of the proventricular nematodes. Spirurid nematodiasis may be an important cause of mortality in young hornbills.

Blockade of cytosolic phospholipase A2 alpha prevents experimental autoimmune encephalomyelitis and diminishes development of Th1 and Th17 responses.

Cytosolic phospholipase A2 alpha (cPLA2 alpha) is the rate-limiting enzyme for release of arachidonic acid, which is converted primarily to prostaglandins via the cyclooxygenase (COX) 1/2 pathways, and leukotrienes via the 5-lipoxygenase (LO) pathway. We utilized inhibitors of cPLA2 alpha, COX-1/2 and 5-LO to determine the potential roles of these enzymes in development of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Blocking cPLA2 alpha prevented EAE development and greatly reduced antigen-induced production of Th1-type cytokines and IL-17. Blocking COX-1/2 delayed onset and reduced severity of EAE, and reduced production of Th1-type cytokines, but not IL-17. Blocking 5-LO delayed onset and reduced cumulative severity of EAE, but did not reduce production of Th1-type cytokines or IL-17. Finally, blockade of cPLA2 alpha from the onset of clinical EAE reduced duration of EAE relapses. Therefore, cPLA2 alpha represents a potential therapeutic target for treatment of MS.

Ranavirus infection of free-ranging and captive box turtles and tortoises in the United States.

Iridoviruses of the genus Ranavirus are well known for causing mass mortality events of fish and amphibians with sporadic reports of infection in reptiles. This article describes five instances of Ranavirus infection in chelonians between 2003 and 2005 in Georgia, Florida, New York, and Pennsylvania, USA. Affected species included captive Burmese star tortoises (Geochelone platynota), a free-ranging gopher tortoise (Gopherus polyphemus), free-ranging eastern box turtles (Terrapene carolina carolina), and a Florida box turtle (Terrepene carolina bauri). Evidence for Ranavirus infection was also found in archived material from previously unexplained mass mortality events of eastern box turtles from Georgia in 1991 and from Texas in 1998. Consistent lesions in affected animals included necrotizing stomatitis and/or esophagitis, fibrinous and necrotizing splenitis, and multicentric fibrinoid vasculitis. Intracytoplasmic inclusion bodies were rarely observed in affected tissues. A portion of the major capsid protein (MCP) gene was sequenced from each case in 2003-2005 and found to be identical to each other and to Frog virus 3 (FV3) across 420 base pairs. Ranavirus infections were also documented in sympatric species of amphibians at two locations with infected chelonians. The fragment profiles of HindIII-digested whole genomic DNA of Ranavirus, isolated from a dead Burmese star tortoise and a southern leopard frog (Rana utricularia) found nearby, were similar. The box turtle isolate had a low molecular weight fragment that was not seen in the digestion profiles for the other isolates. These results suggest that certain amphibians and chelonians are infected with a similar virus and that different viruses exist among different chelonians. Amphibians may serve as a reservoir host for susceptible chelonians. This report also demonstrated that significant disease associated with Ranavirus infections are likely more widespread in chelonians than previously suspected.

Introduction and Establishment of Raccoon Rabies on Islands: Jekyll Island, Georgia, USA as a Case Study.

The introduction of rabies virus (RABV) to barrier islands, which are often popular tourist destinations with resource-rich habitats and connectivity and proximity to the mainland, is especially concerning because it can easily become endemic due to factors like dense rabies-vector populations (e.g., raccoons [ Procyon lotor]), high inter- and intraspecies contact rates, and anthropogenic activities such as supplemental feeding of feral cats ( Felis catus). In January 2013, a neurologic raccoon found on the Jekyll Island (JI), Georgia, US causeway tested positive for rabies. Mortality investigations of 29 raccoons have been conducted between December 2012-May 2017. The two most common diagnoses were RABV ( n=11) and canine distemper virus (CDV; n=8). Parvoviral enteritis was diagnosed in four raccoons but no coinfections were diagnosed. There was no apparent seasonality for rabies cases, but all CDV cases occurred in spring-fall. Most (64%) rabies submissions came from residential or recreational use areas located near feral cat feeding stations. Jekyll Island is a popular destination where tourists engage in numerous outdoor activities which facilitate human-wildlife interactions. Concerns regarding public and animal health highlight the importance of rabies surveillance, prevention, and control on islands. This is the first report of rabies on JI and emphasizes the importance of disease investigations because the assumption that neurologic raccoons have CDV, an endemic pathogen, can miss the establishment of novel pathogens such as RABV.

Evaluation of an endoscopic liver biopsy technique in green iguanas.

OBJECTIVE: To establish a safe and effective endoscopic technique for collection of liver biopsy specimens from lizards by use of a 2.7-mm rigid endoscope system that is commonly available in zoologic veterinary practice. DESIGN: Prospective study. ANIMALS: 11 subadult male green iguanas (Iguana iguana). PROCEDURES: Each lizard was anesthetized, and right-sided coelioscopic examination of the right liver lobe and gallbladder was performed. Three liver biopsy specimens were collected from each lizard by use of a 2.7-mm rigid endoscope and 1.7-mm (5-F) biopsy forceps. Biopsy samples were evaluated histologically for quality and crush artifact. Ten days following surgery, all iguanas were euthanatized and underwent full necropsy examination. RESULTS: For all 11 iguanas, the right liver lobe and gallbladder were successfully examined endoscopically, and 3 biopsy specimens of the liver were collected without complications. Mean +/- SD durations of anesthesia and surgery were 24 +/- 7 minutes and 6.8 +/- 1.0 minutes, respectively. At necropsy, there was no evidence of trauma or disease associated with the skin or muscle entry sites, liver, or any visceral structures in any iguana. All 33 biopsy specimens were considered acceptable for histologic interpretation; in most samples, the extent of crush artifact was considered minimal. CONCLUSIONS AND CLINICAL RELEVANCE: By use of a 2.7-mm rigid endoscope, liver biopsy procedures can be performed safely, swiftly, and easily in green iguanas. Biopsy specimens obtained by this technique are suitable for histologic examination. For evaluation of the liver and biopsy specimen collection in lizards, endoscopy is recommended.

Variation among pathologists in histologic grading of canine cutaneous mast cell tumors.

Ten veterinary pathologists at 1 veterinary institution independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors (MCTs). There was significant variation among pathologists in grading the MCTs (P < 0.001). The probability of assigning a low grade was significantly higher for the pathologists in this study who use a published reference for histologic grading of canine cutaneous MCTs that allows subcutaneous MCTs or MCTs with mitotic figures to be included in the low-grade category (P < 0.0001 and P < 0.0001, respectively).

Renal evaluation in the healthy green iguana (Iguana iguana): assessment of plasma biochemistry, glomerular filtration rate, and endoscopic biopsy.

Plasma biochemistry, iohexol clearance, endoscopic renal evaluation, and biopsy were performed in 23 clinically healthy 2-yr-old green iguanas (Iguana iguana). Mean (+/- SD) values for packed cell volume (30 +/- 3%), total protein (62 +/- 7 g/L, 6.2 +/- 0.7 g/dl), albumin (25 +/- 2 g/L, 2.5 +/- 0.2 g/dl), globulin (37 +/- 6 g/L, 3.7 +/- 0.6 g/ dl), total calcium (3.0 +/- 0.2 mmol/L, 12.0 +/- 0.7 mg/dl), ionized calcium (1.38 +/- 0.1 mmol/L), phosphorus (1.32 +/- 0.28 mmol/L, 4.1 +/- 0.9 mg/dl), uric acid (222 +/- 100 micromol/L, 3.8 +/- 1.7 mg/dl), sodium (148 +/- 3 mmol/L or mEq/ L), and potassium (2.6 +/- 0.4 mmol/L or mEq/L) were considered within normal limits. Values for urea were low (< 1.4 mmol/L, < 4 mg/dl) with 70% of samples below the detectable analyzer range. After the i.v. injection of 75 mg/ kg iohexol into the caudal (ventral coccygeal or tail) vein, serial blood collections were performed over 32 hr. Iohexol assays by high-performance liquid chromatography produced plasma iohexol clearance graphs for each lizard. A three-compartment model was used to fit area under the curve values and to obtain the glomerular filtration rate (GFR) using regression analysis. The mean GFR (SD) was 16.56 +/- 3.90 ml/kg/hr, with a 95% confidence interval of 14.78-18.34 ml/kg/hr. Bilateral endoscopic renal evaluation and biopsy provided tissue samples of excellent diagnostic quality, which correlated with tissue harvested at necropsy and evaluated histologically. None of the 23 animals demonstrated any adverse effects of iohexol clearance or endoscopy. Recommended diagnostics for the evaluation of renal function and disease in the green iguana include plasma biochemical profiles, iohexol clearance, endoscopic examination, and renal biopsy.

Comparison and verification of quantitative competitive reverse transcription polymerase chain reaction (QC-RT-PCR) and real time RT-PCR for avian leukosis virus subgroup J.

Avian leukosis virus subgroup J (ALV-J) infections cause significant economic losses because of increased mortality, tumor production, decreased production, and cost for eradication. Current quantification methods for ALV-J expressed by TCID(50) are difficult to determine because of the lack of cytopathic effect in cell cultures and non-specificity of currently available antigen-capture ELISA tests. In this study, a one-tube fluorescent probe based real time RT-PCR method was developed for quantification of ALV-J and compared with available quantification methods. Cell lysates with different TCID(50)s determined by cell culture and antigen capture ELISA (ag-ELISA) were used for one-tube real time RT-PCR using fluorogenic probe and quantitative competitive RT-PCR (QC-RT-PCR). The results of QC-RT-PCR and real time RT-PCR were highly correlated to the TCID(50)s determined by conventional culture methods. They were also very specific, sensitive, easy to perform, reproducible, and rapid compared with conventional methods. These RT-PCR based quantification methods of ALV-J viral RNA will be useful for virological and pathogenesis studies.

The effects of two endotracheal tube cuff inflation pressures on liquid aspiration and tracheal wall damage in horses.

OBJECTIVE: To investigate the effect of endotracheal tube cuff inflation pressure on the occurrence of liquid aspiration and tracheal wall damage. STUDY DESIGN: Prospective, randomized experimental study. ANIMALS: Ten healthy horses, weighing 535 +/- 55 kg. METHODS: Horses were anesthetized, orotracheally intubated, placed in dorsal recumbency, and maintained on isoflurane in oxygen with controlled ventilation for 175 +/- 15 minutes. The horses were randomly assigned to an endotracheal cuff pressure of 80-100 or 120 cm H2O. The cuff pressure was continuously monitored and maintained at a constant pressure. Methylene blue in saline was instilled proximal to the cuff. After euthanasia, the trachea was opened distal to the endotracheal tube tip to check for evidence of dye leaking past the cuff. The cervical trachea was then resected and opened longitudinally for gross and histologic examinations. RESULTS: No blue staining was found distal to the cuff in any horse. Visual examination of the tracheal mucosa revealed hyperemic and hemorrhagic lesions at the site of the cuff contact. Histologic changes included epithelium attenuation or erosion, submucosal neutrophilic infiltration, and submucosal hemorrhages. Lesions were absent or less extensive in the lower cuff pressure group as compared to the high cuff pressure group. CONCLUSIONS: The endotracheal tube cuff produced a seal sufficient to prevent leakage in both groups. Tracheal wall damage was more severe and occurred more frequently in the higher cuff pressure group. CLINICAL RELEVANCE: Tracheal mucosal damage induced by cuff inflation is pressure-dependent. Cuff pressure monitoring is recommended.

Tetraparesis in a cat with fibrocartilaginous emboli.

An 8-year-old cat, with a history of ataxia that progressed to tetraparesis over a 5-day period, was evaluated. A lesion was localized to the sixth cervical (C6) to second thoracic (T2) spinal cord segments based on physical and neurological examination findings. Blood work was unremarkable, as was survey radiography of the thoracic and abdominal cavities. Cerebrospinal fluid analysis showed moderate neutrophilic inflammation. A definitive diagnosis was not made until necropsy, at which time intravascular fibrocartilaginous embolization (FCE) of the cervical spinal cord was identified. This is only the third published report of FCE in the feline species and the first such case involving the cervical spinal cord.