Detection and diagnosis of Panulirus argus virus 1 in captive spiny lobsters using qPCR in conjunction with histopathology and transmission electron microscopy.

Panulirus argus virus 1 (PaV1) is the first and only naturally occurring pathogenic virus described in the Caribbean spiny lobster, Panulirus argus. PaV1 infection in decapod species that commonly co-occur with P. argus, including the spotted spiny lobster Panulirus guttatus, has not been previously described. In 2016, 14 Caribbean and 5 spotted spiny lobsters were collected near Summerland Key, Florida, to supplement the resident population of the Audubon Aquarium of the Americas in New Orleans, Louisiana. After 5 months in quarantine, Caribbean and spotted spiny lobsters began to exhibit clinical signs of lethargy and dying in the molt. Initial histologic evaluation revealed intranuclear inclusion bodies in circulating hemocytes in the spongy connective tissue of the epidermis, suggesting a viral infection. Samples of hepatopancreas and hemolymph from deceased Caribbean and spotted spiny lobsters tested negative for white spot syndrome virus and positive for PaV1 using real-time quantitative polymerase chain reaction (qPCR). Intranuclear, eosinophilic to amphophilic, Cowdry type A inclusion bodies observed primarily within fixed phagocytes and circulating hemocytes in the hepatopancreas of freshly euthanized Caribbean spiny lobsters were consistent with PaV1 infection. Transmission electron microscopy revealed that hemocytes associated with hepatopancreatic tubules contained viral inclusions with location, size, and morphology consistent with previously described PaV1 infection. These findings highlight the significance of using molecular diagnostics in conjunction with histopathology and electron microscopy in the investigation and diagnosis of PaV1 in spiny lobsters. Further study is required to investigate the relationship of PaV1-associated mortality events and microscopic lesions in the spotted spiny lobster.

Neoplastic signatures: Comparative proteomics of canine hepatobiliary neuroendocrine tumors to normal niche tissue.

Hepatobiliary neuroendocrine neoplasms are rare cancers in humans and dogs. To date, no large-scale primary hepatobiliary neoplasm omics analyses exist in any species. This limits the development of diagnostic biomarkers and targeted therapeutics. Neuroendocrine cancers are a heterogenous group of neoplasms categorized by their tissue-of-origin. Because the anatomic niche of neuroendocrine neoplasms shapes tumor phenotype, we sought to compare the proteomes of 3 canine hepatobiliary neoplasms to normal hepatobiliary tissue and adrenal glands with the objective of identifying unique protein signatures. Protein was extracted from formalin-fixed paraffin-embedded samples and submitted for tandem mass spectroscopy. Thirty-two upregulated and 126 downregulated differentially expressed proteins were identified. Remarkably, 6 (19%) of the upregulated proteins are correlated to non-hepatobiliary neuroendocrine neoplasia and 16 (50%) are functionally annotated within the exosome cellular compartment key to neuroendocrine signaling. Twenty-six (21%) downregulated proteins are enriched in metabolic pathways consistent with alterations in cancer. These results suggests that characteristic neoplastic protein signatures can be gleaned from small data sets using a comparative proteomics approach.

Verminous meningoencephalomyelitis in a red kangaroo associated with Angiostrongylus cantonensis infection.

Angiostrongylus cantonensis is a zoonotic parasitic helminth that normally resides in the pulmonary arteries and the right ventricle of rats (Rattus sp.), the definitive host, where it causes little disease. Humans, dogs, opossums, and various zoo animals are "accidental" hosts. Here we report verminous meningoencephalomyelitis caused by A. cantonensis in a 9-mo-old male red kangaroo (Macropus rufus). The kangaroo was first presented lethargic, recumbent, and hypothermic, with severe muscle wasting. Within 3 wk, he progressed to non-ambulatory paraparesis and died. Gross examination revealed multifocal areas of dark-brown discoloration, malacia, and cavitation in the brain and the spinal cord. Histologically, there were several sections of nematodes surrounded by extensive areas of rarefaction, hemorrhage, spongiosis, neuronal necrosis, and gliosis. Based on size, morphology, and organ location, the nematodes were identified as subadult males and females. Interestingly, an eosinophilic response was largely absent, and the inflammatory response was minimal. A. cantonensis infection had not been reported previously in a red kangaroo in Louisiana or Mississippi, to our knowledge. Our case reaffirms the widespread presence of the helminth in the southeastern United States and indicates that A. cantonensis should be considered as a differential in macropods with neurologic clinical signs in regions where A. cantonensis is now endemic.

Locally Transmitted Trypanosoma cruzi in a Domestic Llama (Lama glama) in a Rural Area of Greater New Orleans, Louisiana, USA.

Trypanosoma cruzi-associated megaesophagus was diagnosed in a domestic Louisiana-born llama with no significant travel history. The llama resided in the same rural area of greater New Orleans, Louisiana, where the first human autochthonous case of Chagas disease was identified in the state. Venous blood from the llama tested positive for T. cruzi kinetoplastid DNA by conventional PCR. The cardiac evaluation was unremarkable, while thoracic radiographs revealed generalized megaesophagus. The llama received supportive care, but was ultimately humanely euthanized. The esophagus was severely distended throughout its length on necropsy, and histologic evaluation showed no microscopic changes in esophageal tissue and minimal to mild lymphoplasmacytic inflammation in cardiac tissue. T. cruzi DNA was detected by conventional PCR in the esophagus, small intestine, and blood despite no protozoan organisms being observed in multiple tissue sections examined. This report contributes to the growing body of evidence of local transmission of T. cruzi in the southern United States, and Chagas disease should be considered a differential diagnosis when evaluating llamas and other large animal species for esophageal dysfunction. There is little research describing megaesophagus or Chagas disease in llamas, and this report aims to increase awareness about this zoonotic disease that is becoming more frequently reported in the southern United States.

Pathologic and immunohistochemical findings in an outbreak of systemic toxoplasmosis in a mob of red kangaroos.

Toxoplasma gondii is a zoonotic protozoan pathogen that infects many endothermic vertebrates, including humans; the domestic cat and other felids serve as the definitive host. Macropodids are considered highly susceptible to toxoplasmosis. Here, we describe the clinical, pathologic, and immunohistochemical findings of an outbreak of systemic toxoplasmosis in a mob of 11 red kangaroos (Macropus rufus), with high morbidity (73%) and mortality (100%) rates. Affected animals had either severe and rapidly deteriorating clinical conditions or sudden death, which was correlated with widespread necrotizing lesions in multiple organs and intralesional T. gondii organisms identified via MIC3-specific immunohistochemistry and confirmed by REP529-specific rtPCR. Quantification of parasites demonstrated the highest parasite density in pulmonary parenchyma compared with other tissues. Our study highlights the continued importance of this severe condition in Australian marsupials.

Neurologic amebiasis caused by Balamuthia mandrillaris in an Indian flying fox (Pteropus giganteus).

A 4-5-month-old intact male Indian flying fox (Pteropus giganteus) was presented to the Baton Rouge Zoo's veterinary hospital with an acute onset of obtundation that was diagnosed with amebic encephalitis. Histologic examination revealed numerous amebic trophozoites within necrotic foci, affecting the occipital cerebrum and surrounding the mesencephalic aqueduct. The etiologic agent, Balamuthia mandrillaris, was determined by multiplex quantitative real-time polymerase chain reaction, immunohistochemistry, and indirect fluorescent antibody test. The current report documented a case of amebic encephalitis within the order Chiroptera.

Fungal myocarditis and pericardial effusion secondary to Inonotus tropicalis (phylum Basidiomycota) in a dog.

Fungal disease is a rare cause of pericardial effusion in dogs. This report describes the first case of fungal pericardial effusion and myocarditis secondary to the fungal organism Inonotus tropicalis. A 9-year-old female spayed French bulldog with a multi-year history of treatment with glucocorticoids for management of atopy was presented for exercise intolerance, ascites and weight loss. Physical examination and thoracic imaging revealed enlarged peripheral and cranial mediastinal lymph nodes, left ventricular thickening and cardiac tamponade secondary to pericardial effusion. Fine needle aspiration of the cranial mediastinal lymph node showed pyogranulomatous inflammation with short, thin and poorly septated hyphae. Culture of the aspirate yielded a fungal isolate identified as Inonotus tropicalis based on morphologic features and rRNA gene sequencing. Postmortem examination showed myocardial thickening with multifocal to coalescing, firm, white, ill-defined nodules. Histology confirmed the presence of disseminated fungal infection with extensive myocardial involvement. Inonotus tropicalis is an opportunistic poroid wood-decaying basidiomycete. Infection in this dog was likely the result of chronic immunosuppressive therapy.

Long-term management of ovarian neoplasia in two cockatiels (Nymphicus hollandicus).

Cockatiels (Nymphicus hollandicus) are commonly diagnosed with ovarian neoplasia. However, there is very little information regarding medical management of this disease condition and subsequent patient response. Long-term medical therapy of 2 cockatiels eventually diagnosed with ovarian neoplasia is described along with responses to the treatment regimens. Each bird had initial signs consistent with reproductive disease (chronic egg laying, ascites, and lethargy) and respiratory distress. The diagnosis of ovarian adenocarcinoma was confirmed on postmortem examination of both birds. The birds were conservatively managed by periodic coelomocentesis and gonadotropin-releasing hormone (GnRH) agonist administration for 9 and 25 months, respectively. A positive response to GnRH agonist therapy was documented in 1 of the 2 birds. These 2 cases demonstrate that periodic coelomocentesis with or without GnRH agonist therapy may be a viable option for the long-term management of ovarian neoplasia and reproductive-organ-associated ascites in cockatiels.

Moh's micrographic surgery for the management of a periocular mast cell tumor in a dog.

A 3-year-old neutered male boxer dog presented with a 6-month history of a waxing and waning mass of the left dorsotemporal eyelid margin. Cytology and biopsy confirmed a diagnosis of mast cell neoplasia. Systemic staging of the dog failed to reveal any evidence of metastatic neoplasia. Owing to the location of the tumor within the eyelid margin and the wide surgical margins recommended for excision of mast cell tumors, Mohs micrographic surgery (MMS) was chosen for its potential to conserve tissues while providing intraoperative confirmation the tumor was completely excised. Utilizing MMS horizontal sectioning technique, 100% of the surgical margins were assessed prior to closure of the surgical wound. This represents the first time a comprehensive MMS protocol was used in a veterinary patient under general anesthesia.

Cutaneous epitheliotropic lymphoma with dual CD3 and c-kit expression in a dog.

An 11-year-old 8.9-kg spayed female Boston Terrier was presented for evaluation of a mucocutaneous tumor on the right side of the upper lip that had been biopsied (punch biopsy) by the referring veterinarian. The histologic diagnosis was poorly differentiated round cell tumor involving the submucosa with patchy involvement of the mucosa. On presentation of the dog to Louisiana State University, the tumor was found to involve the mucosa and haired skin surface of the right upper lip. A fine-needle aspirate of the right mandibular lymph node contained atypical poorly differentiated round cells similar to those in the histologic sections. To further characterize the tumor, immunohistochemical analysis of the tumor on the lip was performed; tumor cells were strongly immunoreactive for both CD3 and c-kit in a cytoplasmic to membranous pattern, with CD3 expression having a more intense membranous component. The diagnosis was cutaneous epitheliotropic T-cell lymphoma with co-expression of CD3 and c-kit by neoplastic lymphocytes, an unusual finding. As receptor tyrosine kinases can be attractive targets for cancer treatment, expression of these molecular targets in tumors is a promising subject of future research.

Clenbuterol toxicosis in three Quarter Horse racehorses after administration of a compounded product.

CASE DESCRIPTION: 3 Quarter Horse racehorses were examined for suspected clenbuterol overdose 12 to 24 hours after administration by mouth of a compounded clenbuterol product. CLINICAL FINDINGS: All horses developed sinus tachycardia, muscle tremors, hyperhidrosis, and colic. Abnormalities on serum biochemical analysis included hyperglycemia, azotemia, and high creatine kinase activity. The presence of clenbuterol in the serum of all 3 horses and in the product administered was confirmed and quantified by use of liquid chromatography-electrospray tandem mass spectrometry. TREATMENT AND OUTCOME: Propranolol (0.01 mg/kg [0.005 mg/lb], IV) was administered to all 3 horses for antagonism of ß-adrenergic effects and caused a transient decrease in heart rate in all patients. All horses also received crystalloid fluids IV and other supportive treatment measures. Two horses were euthanatized (2 and 4 days after admission) because of complications. One horse recovered and was discharged 4 days after admission to the hospital. In the 2 nonsurviving horses, skeletal and cardiac muscle necrosis was evident at necropsy, and tissue clenbuterol concentrations were highest in the liver. CLINICAL RELEVANCE: Clenbuterol is a ß(2)-adrenergic receptor agonist licensed for veterinary use as a bronchodilator. At doses ≥ 10² µg/kg (4.5 µg/lb), in excess of those normally prescribed, ß-adrenergic stimulation by clenbuterol may cause sustained tachycardia, muscle tremors, hyperglycemia, and cardiac and skeletal muscle necrosis. Laminitis, acute renal failure, rhabdomyolysis, and cardiomyopathy were fatal complications associated with clenbuterol overdose in 2 horses in the present report. At the dose administered, propranolol was effective for short-term control of sinus tachycardia, but it did not alleviate all clinical signs in patients in the present report. These cases demonstrated the risks associated with the use of nonprescribed compounded medications for which the ingredients may be unknown.

Idiopathic solitary cutaneous xanthoma in a dog.

A 6-year-old female spayed Boxer mix dog was presented with multiple cutaneous masses, one of which was determined to be a xanthoma. Fine-needle aspirates of this mass revealed large round cells that were consistent with macrophages. These macrophages had lightly basophilic cytoplasm that was filled with many clear circular spaces that varied in size. The nuclei of these cells displayed mild anisokaryosis with condensed chromatin and lacked prominent nucleoli. The cytologic interpretation was lipid-laden histiocytic inflammation most consistent with a cutaneous xanthoma, which was confirmed histologically. Mild hypertriglyceridemia and persistent moderate hypercholesterolemia were present. After ruling out other causes of hyperlipidemia, we concluded that the dog likely had idiopathic hyperlipidemia with secondary xanthoma formation.

Intermittent claudication-like syndrome secondary to atherosclerosis in a yellow-naped Amazon parrot (Amazona ochrocephala auropalliata).

A 25-year-old yellow-naped Amazon parrot (Amazona ochrocephala auropalliata) was presented for nasal discharge and sneezing. Physical examination revealed poor feather quality, a mild serous nasal discharge, and a mass on the dorsal surface of the oral cavity. Cytologic examination of a mass aspirate as well as results of a choanal culture revealed squamous metaplasia of the salivary glands and bacterial rhinitis, respectively. Following resolution of the presenting conditions, the patient was presented for hind limb weakness and ataxia. The clinical signs were transient and generally resolved with rest but could be reproduced after stressful episodes, such as restraint for procedures or treatment. Test results from a complete blood count, biochemistry profile, whole-body radiographs, needle electromyography of the leg muscles, and an edrophonium challenge test were within reference limits. Based on the clinical signs and results of the diagnostic workup, the presumptive diagnosis was intermittent claudication, a condition caused by peripheral vascular disease and defined as intermittent weakness and pain in the legs induced by exercise and relieved by rest. Shortly after initiation of treatment with isoxsuprine, the bird died. Postmortem examination and histopathology revealed severe atherosclerotic lesions throughout the vascular system with stenotic lesions present in the abdominal aorta and femoral arteries. Electron microscopic examination of the great arteries was also performed and helped to further characterize the nature of the lesions. This case is the first report, to our knowledge, of an intermittent claudication-like syndrome associated with peripheral atherosclerosis in a psittacine bird. In addition, the distribution and some of the macroscopic and histopathologic features of the lesions differ from previous descriptions of atherosclerosis in psittacine birds.