Nicholas A. Crossland

Nannizziopsis guarroi infection in 2 Inland Bearded Dragons (Pogona vitticeps): clinical, cytologic, histologic, and ultrastructural aspects

Chrysosporium-related infections have been increasingly reported in reptiles over the last 2 decades. In this report, we describe clinical, cytologic, histopathologic, and ultrastructural aspects of Chrysosporium-related infection in 2 Inland Bearded Dragons (Pogona vitticeps). Case 1 was presented for an enlarging raised lesion over the left eye and multiple additional masses over the dorsum. Case 2 was submitted to necropsy by the referring veterinarian for suspected yellow fungus disease. Impression smears of the nodules in case 1 revealed granulomatous to pyogranulomatous inflammation and many septate, variably long, 4-10 μm wide, often undulated hyphae, and very rare conidia. Postmortem impression smears of the superficial lesions of case 2 contained large numbers of solitary conidia and arthroconidia and low numbers of hyphae with similar morphology to case 1. Histopathology of the 2 cases revealed severe, multifocal, chronic, ulcerative, nodular pyogranulomatous dermatitis, with myriad intralesional septate hyphae, and arthroconidia. Fungal culture and molecular sequencing in both cases indicated infection with Nannizziopsis guarroi.

Late Disseminated Lyme Disease: Associated Pathology and Spirochete Persistence Posttreatment in Rhesus Macaques

Nonhuman primates currently serve as the best experimental model for Lyme disease because of their close genetic homology with humans and demonstration of all three phases of disease after infection with Borrelia burgdorferi. We investigated the pathology associated with late disseminated Lyme disease (12 to 13 months after tick inoculation) in doxycycline-treated (28 days; 5 mg/kg, oral, twice daily) and untreated rhesus macaques. Minimal to moderate lymphoplasmacytic inflammation, with a predilection for perivascular spaces and collagenous tissues, was observed in multiple tissues, including the cerebral leptomeninges, brainstem, peripheral nerves from both fore and hind limbs, stifle synovium and perisynovial adipose tissue, urinary bladder, skeletal muscle, myocardium, and visceral pericardium. Indirect immunofluorescence assays that combined monoclonal (outer surface protein A) and polyclonal antibodies were performed on all tissue sections that contained inflammation. Rare morphologically intact spirochetes were observed in the brains of two treated rhesus macaques, the heart of one treated rhesus macaque, and adjacent to a peripheral nerve of an untreated animal. Borrelia antigen staining of probable spirochete cross sections was also observed in heart, skeletal muscle, and near peripheral nerves of treated and untreated animals. These findings support the notion that chronic Lyme disease symptoms can be attributable to residual inflammation in and around tissues that harbor a low burden of persistent host-adapted spirochetes and/or residual antigen.

EFFECTS OF PLANT PROTEIN DIETS ON THE HEALTH OF FARMED AMERICAN ALLIGATORS (ALLIGATOR MISSISSIPPIENSIS)

Abstract The objective of this prospective, blinded study was to compare plasma biochemical values and gross and histologic evaluation of kidney and liver from American alligators (Alligator mississippiensis) fed extruded diets with protein derived from animal or plant sources. Alligators in two treatment groups were fed an extruded diet with protein derived primarily from plant products for 7 (n = 20) or 10 (n = 20) mo prior to harvest. A control group (n = 20) was fed a commercial diet with protein derived from animal products for the duration of the study. Plasma biochemistry panels were obtained and gross and histologic examination of kidney and liver tissues was conducted for each animal. No differences were found between alligators fed diets with animal or plant protein in terms of either biochemistry profiles or gross or histologic examination of kidney and liver. Plant-based diets, fed for up to 10 mo, do not appear to have any ill effects on the kidney or liver of American alligators.

First report of Cylicospirura felineus in a feral domestic shorthair cat in North America

Case summary A feral domestic shorthair cat was euthanized owing to acute onset and progression of neurological signs attributed to ethylene glycol toxicity. At post-mortem examination two nodules were identified within the fundus of the stomach. Examination of the gastric nodules revealed an intact mucosal surface, each with multiple red slender nematodes extending through an individual central pore. Histopathological evaluation of the nodules highlighted unique reactive fibroplasia, mimicking feline gastrointestinal eosinophilic sclerosing fibroplasia (FGESF), encasing numerous nematodes with females possessing gravid uteri containing abundant larvated eggs. The latter findings were highly suggestive of the Cylicospirura genus, further supported by an en face evaluation of the buccal cavity, highlighting a distinctive trifid tooth appearance. Together, these findings are consistent with Cylicospirura felineus. PCR for the COX-1 gene was unsuccessful on formalin-fixed specimens, attributed to nucleic acid and protein crosslinking. Relevance and novel information This represents the first documented case of Cylicospirura species in a feral domestic shorthair cat in North America. This particular cat lived in the highly urban environment of New Orleans, Louisiana. Identification of this case demonstrates the potential for feral cats to serve as reservoir hosts and ultimately support transmission of Cylicospirura species into domesticated cat populations. Gastric cylicospiruriasis may present clinically as a firm abdominal mass, potentially with a history of chronic vomiting. The latter emphasizes the importance of differentiating this condition from a neoplastic process such as alimentary lymphoma and adenocarcinoma. Histologically, the unique thick anastomosing collagenous cords encasing nematodes represent a stereotypical response observed in a broad array of gastrointestinal inflammation in felines, including intralesional bacteria, fungal hyphae, foreign bodies and, in this case, gastric nematodes that closely resemble FGESF. Additionally, these unique histological lesions have previously been misinterpreted as neoplastic conditions, including sclerosing mast cell tumor and extraosseous osteosarcoma.

Pneumonia in a Captive Central Bearded Dragon With Concurrent Detection of Helodermatid Adenovirus 2 and a Novel Mycoplasma Species

A 4-year-old captive male central bearded dragon (Pogona vitticeps) was presented for recurrent episodic dyspnea and anorexia with occasional expulsion of oral mucoid discharge. Despite empirical antimicrobial therapy and supportive care, the animal died and was submitted for autopsy. Defining histologic features included heterophilic and lymphocytic interstitial pneumonia, with occasional amphophilic intranuclear inclusions and prominent type II pneumocyte hyperplasia. Transmission electron microscopy revealed intranuclear 80-nm, nonenveloped, hexagonal viral particles within pneumocytes. Helodermatid adenovirus 2 (HeAdV2) was determined as the etiologic agent through pan-adenoviral consensus polymerase (PCR) chain reaction and sequencing. Nucleic acid from a novel Mycoplasma sp. (provisionally called Mycoplasma pogonae) was identified by pan-generic PCR targeting the mycoplasma 16S ribosomal RNA gene with sequencing and phylogenetic analysis. As bacteria morphologically consistent with Mycoplasma sp. were not observed by special stains and transmission electron microscopy, the detection of M. pogonae nucleic acid is of indeterminate significance; however, M. pogonae and HeAdV2 coinfection may have exacerbated disease.

Investigation of a Cyprinid Herpesvirus 1 Disease Episode in a Group of Pond-Reared Koi

Fifteen adult koi (a variant of Common Carp Cyprinus carpio) simultaneously developed white cutaneous proliferations affecting up to 30% of their bodies. The onset of these lesions (in September 2014) was associated with their return to a remodeled backyard water garden after temporarily being maintained in a plastic swimming pool. A single water temperature taken during the outbreak read 21°C on November 17, 2014. The water garden had no extrinsic heat source, with average ambient temperatures ranging from 9.4 to 26.4°C during the outbreak (September 2014-January 2015). Representative skin biopsies were obtained from two fish; the histologic features included severe epidermal hyperplasia, dysplasia, keratinocyte apoptosis, decreased and haphazardly distributed goblet cells with the absence of club cells, keratinocyte hydropic degeneration, and moderate infiltration by lymphocytes and eosinophilic granular cells. Ultrastructural findings included intranuclear nonenveloped hexagonal nucleocapsids and abundant cytoplasmic-enveloped virions morphologically consistent with the Alloherpesviridae family. Polymerase chain reaction was performed on formalin-fixed, paraffin-embedded shavings from the two biopsied koi targeting the thymidine kinase gene of cyprinid herpesvirus 1 (CyHV-1). Together with the aforementioned findings, these results are consistent with an outbreak of CyHV-1 in a population of adult koi.

Aberrant Migration of Cuterebra Larvae in 2 Domestic Rabbits (Oryctolagus cuniculus)

Abstract This report describes 2 cases of cuterebriasis in an atypical host, the domestic rabbit (Oryctolagus cuniculus). Case 1 describes the clinical presentation, diagnosis and management of ophthalmomyiasis externa caused by a bot fly larva. Case 2 outlines the clinical syndromes, postmortem, and histopathological findings associated with Cuterebra larval migrans through the respiratory and central nervous systems. In both cases the presence of the parasite caused significant debilitating disease in the host animals, with fatal consequences in one patient.

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.