The first complete papillomavirus genome characterized from a marsupial host: a novel isolate from Bettongia penicillata.

The first fully sequenced papillomavirus (PV) of marsupials, tentatively named Bettongia penicillata papillomavirus type 1 (BpPV1), was detected in papillomas from a woylie (Bettongia penicillata ogilbyi). The circular, double-stranded DNA genome contains 7,737 bp and encodes 7 open reading frames (ORFs), E6, E7, E1, E2, E4, L2, and L1, in typical PV conformation. BpPV1 is a close-to-root PV with L1 and L2 ORFs most similar to European hedgehog PV and bandicoot papillomatosis carcinomatosis virus types 1 and 2 (BPCV1 and -2). It appears that the BPCVs arose by recombination between an ancient PV and an ancient polyomavirus more than 10 million years ago.

Clinical chemistry values and tissue enzyme activities in western barred bandicoots (Perameles bougainville).

BACKGROUND: The western barred bandicoot, Perameles bougainville, is an endangered Australian marsupial species whose survival is threatened by a papillomatosis and carcinomatosis syndrome. Investigations to characterize this syndrome would benefit from species-specific clinical chemistry data. OBJECTIVES: The purpose of this study was to determine plasma biochemical reference values and to determine enzyme activities in various tissues of P. bougainville. METHODS: Heparinized blood samples were collected by jugular venipuncture from 53 clinically healthy bandicoots of both sexes and at 3 geographic locations. Plasma was analyzed for routine clinical chemistry variables using an automated biochemistry analyzer. Tissues obtained following humane euthanasia of 3 bandicoots were analyzed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine kinase (CK), alpha-amylase (AML), and gamma-glutamyltransferase (GGT) activities. RESULTS: Significant differences in the results were found for animals based on geographic location and sex; hence, results were expressed as minimum and maximum values. A population reference interval was calculated for AST activity (20-283 U/L). ALT was found mainly in the liver, with lower levels in cardiac and skeletal muscle and kidneys. AST was detectable in many tissues, including the heart, liver, kidneys, and central nervous system; CK was found in skeletal and cardiac muscle and central nervous system; AML was found in the pancreas; and GGT was found mainly in kidneys with lower levels in the intestines and pancreas. CONCLUSIONS: These findings will facilitate the interpretation of clinical chemistry results from P. bougainville and thereby inform population management and clinical decision-making.

Genomic characterization of a novel virus found in papillomatous lesions from a southern brown bandicoot (Isoodon obesulus) in Western Australia.

The genome of a novel virus, tentatively named bandicoot papillomatosis carcinomatosis virus type 2 (BPCV2), obtained from multicentric papillomatous lesions from an adult male southern brown bandicoot (Isoodon obesulus) was sequenced in its entirety. BPCV2 had a circular double-stranded DNA genome consisting of 7277 bp and open reading frames encoding putative L1 and L2 structural proteins and putative large T antigen and small t antigen transforming proteins. These genomic features, intermediate between Papillomaviridae and Polyomaviridae are most similar to BPCV1, recently described from papillomas and carcinomas in the endangered western barred bandicoot (Perameles bougainville). This study also employed in situ hybridization to definitively demonstrate BPCV2 DNA within lesion biopsies. The discovery of BPCV2 provides evidence of virus-host co-speciation between BPCVs and marsupial bandicoots and has important implications for the phylogeny and taxonomy of circular double-stranded DNA viruses infecting vertebrates.

In situ hybridization to detect bandicoot papillomatosis carcinomatosis virus type 1 in biopsies from endangered western barred bandicoots (Perameles bougainville).

The western barred bandicoot (Perameles bougainville) is an endangered Australian marsupial species in which a papillomatosis and carcinomatosis syndrome occurs. Bandicoot papillomatosis carcinomatosis virus type 1 (BPCV1) is associated with the lesions of this progressively debilitating syndrome. Five digoxigenin-labelled DNA probes were generated for in situ hybridization (ISH) and the technique was optimized and performed on formalin-fixed paraffin-embedded (FFPE) biopsies. Staining of keratinocyte and sebocyte nuclei within lesions was achieved with all five probes. The sensitivity of ISH (76.9%) surpassed that of PCR (30.8%) for FFPE samples. The sensitivity of ISH varied from 81% (papillomas) and 70% (carcinoma in situ) to 29% (squamous cell carcinomas). The specificity of the test was confirmed using an irrelevant probe and papillomas from other species. These results strengthen the association between BPCV1 and the western barred bandicoot papillomatosis and carcinomatosis syndrome and give insight into the biology of the virus-host interaction.

Hematologic characteristics of captive western barred bandicoots (Perameles bougainville) from Western Australia.

BACKGROUND: The western barred bandicoot (Perameles bougainville) is an Australian marsupial species now considered endangered as a consequence of habitat destruction and predation. A recently discovered papillomatosis syndrome is hindering efforts to repopulate this species. Hematology reference intervals have been lacking for P bougainville, preventing optimal interpretation of hematology results from wart-affected and clinically normal animals. OBJECTIVES: The purpose of this study was to establish hematology reference values and describe morphologic characteristics of blood cells of healthy western barred bandicoots. METHODS: Fifty-nine whole blood samples were collected by jugular venipuncture into EDTA from 47 clinically healthy captive western barred bandicoots at 3 locations on the Western Australian mainland. A CBC was performed using an ADVIA-120 analyzer. Data were compared on the basis of geographic location, sex, age, and lactation status, and reference intervals were calculated. Blood cell morphology was evaluated using light microscopy, and transmission and scanning electron microscopy. RESULTS: Significant differences were found based on sex (RBC indices, fibrinogen), age (% polychromatophilic RBCs), and geographic location (RBC, neutrophil, and lymphocyte counts, MCHC, % polychromatophilic RBCs, fibrinogen). Combined reference intervals were calculated for hemoglobin concentration (122-165 g/L), HCT (0.36-0.49 L/L), and total WBC (2.9-14.9 x 10(9)/L), monocyte (0-0.6 x 10(9)/L), eosinophil (0-0.9 x 10(9)/L), and total plasma protein (47-63 g/L) concentrations. Leukocyte, erythrocyte, and platelet morphology were similar to those of other marsupial peramelid species. Nuclei in neutrophils, monocytes, and eosinophils occasionally had an annular configuration. CONCLUSIONS: Reference intervals and blood cell morphology obtained in this study will be useful for the evaluation of laboratory data from ill animals and assist with population health monitoring of western barred bandicoots.

A novel virus detected in papillomas and carcinomas of the endangered western barred bandicoot (Perameles bougainville) exhibits genomic features of both the Papillomaviridae and Polyomaviridae.

Conservation efforts to prevent the extinction of the endangered western barred bandicoot (Perameles bougainville) are currently hindered by a progressively debilitating cutaneous and mucocutaneous papillomatosis and carcinomatosis syndrome observed in captive and wild populations. In this study, we detected a novel virus, designated the bandicoot papillomatosis carcinomatosis virus type 1 (BPCV1), in lesional tissue from affected western barred bandicoots using multiply primed rolling-circle amplification and PCR with the cutaneotropic papillomavirus primer pairs FAP59/FAP64 and AR-L1F8/AR-L1R9. Sequencing of the BPCV1 genome revealed a novel prototype virus exhibiting genomic properties of both the Papillomaviridae and the Polyomaviridae. Papillomaviral properties included a large genome size ( approximately 7.3 kb) and the presence of open reading frames (ORFs) encoding canonical L1 and L2 structural proteins. The genomic organization in which structural and nonstructural proteins were encoded on different strands of the double-stranded genome and the presence of ORFs encoding the nonstructural proteins large T and small t antigens were, on the other hand, typical polyomaviral features. BPCV1 may represent the first member of a novel virus family, descended from a common ancestor of the papillomaviruses and polyomaviruses recognized today. Alternatively, it may represent the product of ancient recombination between members of these two virus families. The discovery of this virus could have implications for the current taxonomic classification of Papillomaviridae and Polyomaviridae and can provide further insight into the evolution of these ancient virus families.

Lymphangiosarcoma in two cats.

Lymphangiosarcoma is a malignant neoplasm of the lymphatic endothelium that is rare in cats. This report describes two cases of feline lymphangiosarcoma that originated in the distal limb, causing intractable lymphoedema and serosanguineous discharge with ecchymoses in local and distant sites. In association with the neoplasia, one cat had cortical bone lysis of multiple metacarpal bones of the affected limb and the other had severe immune-mediated haemolytic anaemia (IMHA). The disease in both cases affected young cats and progressed rapidly. Persistent distal limb lymphoedema with serosanguineous discharge is suggestive of lymphangiosarcoma especially when local or distal ecchymoses are evident.

Evaluation of lymphoma in a cat using 99mTc-sestamibi.

This report describes the evaluation of cutaneous lymphoma in a cat using 99mTc-sestamibi. A mass in the left pes with extension to the left popliteal lymph node was diagnosed as B-cell lymphoma. Thoracic and abdominal radiography and abdominal ultrasound were unremarkable. Scintigraphy using 99mTc-sestamibi confirmed the lesion was confined to the left hind leg, and the leg was amputated. The cat made a full recovery.

Gingival eruption cysts induced by cyclosporine administration to neonatal dogs.

OBJECTIVES: The purpose of this paper is to report the development of extensive, gingival eruption cyst formation in neonatal dogs during chronic administration of cyclosporine, which regressed after treatment ceased. MATERIALS AND METHODS: As part of an investigation of the efficacy of adenoviral-mediated gene therapy in a canine model of Duchenne Muscular Dystrophy (DMD), 26 Golden retriever pups were given intramuscular injections of adenoviral gene constructs (at 2-5 days old) either with (15 pups) or without (9 pups) concomitant immunosuppression using oral cyclosporine (from 1 to 3 days old). RESULTS: The daily administration of cyclosporine to Golden Retriever pups from 1 to 3 days of age resulted in the formation of eruption cysts over the incisors after approximately 3 weeks of treatment. The gingiva over the remaining teeth became swollen. However unlike the incisors, part of the crown erupted through the gingiva. Continuous treatment for 60-90 days was associated with persistence and expansion of the cysts over the incisor teeth. The cysts resolved within 1 month if cyclosporine administration was discontinued. Movement of incisors within the jaws was not appreciably affected when compared with age-matched controls. CONCLUSION: Chronic cyclosporine administration to neonatal dogs induces the formation of eruption cysts over the incisors, which is reversible once treatment is discontinued and does not affect intraosseous eruption.