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Mark D. Bennett BVSc (Hons), MACVSc (Veterinary Pathology), PhD

Supervisors

  • Dr Amanda O’Hara BSc (Hons), BVMS, MACVSc, Diplomate ACVP, Murdoch University
  • Dr Philip Nicholls BSc (Hons), BVSc, PhD, MRCPath MRCVS, Murdoch University
  • Dr Kris Warren BSc BVMS (Hons) PhD, Murdoch University

For more than a decade, community groups, scientific organizations and government agencies have collaborated to repopulate the endangered western barred bandicoot (Perameles bougainville). While initially successful, the unexpected discovery of a papillomatosis and carcinomatosis syndrome in captive and wild populations of P. bougainville exposed a dearth of knowledge regarding their diseases. Dr Bennett’s research addressed this issue through study of the clinical pathology, parasitology, immunology and virology of P. bougainville.

To facilitate the detection and understanding of diseases of P. bougainville, guidelines for interpreting haematology and clinical chemistry results were developed. This included calculating species-specific reference intervals for plasma aspartate transaminase activity (20–283 U/L), haemoglobin (122-165 g/L), haematocrit (0.36-0.49 L/L), total leukocyte (2.9-14.9 x109/L), monocyte (0-0.6 x109/L), eosinophil (0-0.9 x109/L) concentrations and total protein (47-63 g/L) estimated by refractometry. P. bougainville immunoglobulin was also fractionated from plasma and inoculated into sheep to derive antiserum for serological screening assays. 

Arthropods, helminths and protozoa parasitic on P. bougainville were catalogued and Eimeria kanyana n. sp. was formally described. The pathogenic and zoonotic potential of bacteria detected in ticks parasitic on P. bougainville was also considered.

The association between bandicoot papillomatosis carcinomatosis virus type 1 (BPCV1) and the western barred bandicoot papillomatosis and carcinomatosis syndrome was investigated using PCR, in situ hybridization and virus isolation. Optimized in situ hybridization techniques demonstrated BPCV1 DNA within keratinocyte and sebocyte nuclei, and BPCV1 mRNA within the cytoplasm. BPCV1 virions were isolated by ultracentifugation and visualized with negative stain transmission electron microscopy, revealing icosahedral, non-enveloped virions ~47 nm in diameter similar to viruses belonging to the Papillomaviridae and Polyomaviridae.

A novel virus, tentatively named bandicoot papillomatosis carcinomatosis virus type 2 (BPCV2) was discovered in papillomatous lesions from a southern brown bandicoot (Isoodon obesulus). It had a circular double-stranded DNA genome of 7277 bp, and encoded two papillomavirus-like structural proteins, L1 and L2, and two polyomavirus-like putative transforming proteins, large T antigen and small t antigen. DNA and RNA in situ hybridization confirmed the presence of BPCV2 nucleic acids within lesion biopsies. The discovery of the bandicoot papillomatosis carcinomatosis viruses has provoked reassessment of the established virus taxonomy paradigm, theories of virus-host co-speciation and bandicoot population management strategies.

This thesis is available in Murdoch University's Digital Thesis collection - http://wwwlib.murdoch.edu.au/adt/

Publications

  1. Bennett, M. D. (2003). Pseudophysaloptera enigmatica sp. nov. (Nematoda: Physalopteridae) from Dactylopsila trivirgata Gray, 1858 (Marsupialia: Petauridae) in Australia, with a review of the genus. Parasite. 10(2):205-210.
  2. Bennett, M. D., L. Woolford, A. J. O’Hara, P. K. Nicholls, K. Warren, R. P. Hobbs (2006). A new Eimeria species parasitic in western barred bandicoots Perameles bougainville (Marsupialia: Peramelidae) in Western Australia. Journal of Parasitology. 92(6):1292-1294.
  3. Bennett, M. D., L. Woolford, A. J. O’Hara, P. K. Nicholls, K. Warren, R. Swan, J. A. Friend (2007).Klossiella quimrensis (Apicomplexa: Klossiellidae) causes renal coccidiosis in western barred bandicoots Perameles bougainville (Marsupialia: Peramelidae) in Western Australia. Journal of Parasitology. 93(1):89-92.
  4. Bennett, M. D. (editor). Proceedings of the western barred bandicoot symposium 2007. Murdoch University, Murdoch, Western Australia, February 2007.
  5. Bennett, M. D., L. Woolford, A. J. O’Hara, P. K. Nicholls, K. Warren, K. L. Hulme-Moir, P. Clark (2007). Hematologic characteristics of captive western barred bandicoots Perameles bougainville (Marsupialia: Peramelidae) from Western Australia. Veterinary Clinical Pathology. 36(4):348-353.
  6. Bennett, M. D., L. Woolford, A. J. O’Hara, K. S. Warren, P. K. Nicholls (2008). In situ hybridization to detect bandicoot papillomatosis carcinomatosis virus type 1 in biopsies from endangered western barred bandicoots (Perameles bougainville). Journal of General Virology. 89:419-423.
  7. Bennett, M. D., L. Woolford, P. K. Nicholls, K. S. Warren and A. J. O’Hara (2008). Hepatic intranuclear glycogen inclusions in western barred bandicoots (Perameles bougainville). Journal of Veterinary Diagnostic Investigation. 20:376-379.
  8. Bennett, M. D., L. Woolford, A. J. O’Hara, P. K. Nicholls, K. S. Warren (2008). Clinical chemistry and tissue enzyme activities in western barred bandicoots (Perameles bougainville). Veterinary Clinical Pathology. 37(2):221-224.
  9. Bennett, M. D., L. Woolford, H. Stevens, M. Van Ranst, T. Oldfield, M. Slaven, A. J. O’Hara, K. S. Warren and P. K. Nicholls (2008). Genomic characterization of a novel virus found in papillomatous lesions from a southern brown bandicoot (Isoodon obesulus) in Western Australia. Virology. 376(1):173-182.
  10. Bennett, M. D. (2008). Western barred bandicoots in health and disease. PhD thesis. Murdoch University.
  11. Bennett, M. D., A. Reiss, H. Stevens, E. Heylen, M. Van Ranst, A. Wayne, M. Slaven, J. N. Mills, K. S. Warren, A. J. O’Hara and P. K. Nicholls (2010). The first complete papillomavirus genome characterized from a marsupial host: a novel isolate from Bettongia penicillata. Journal of Virology. 84(10):5448-5453.

Publications as a co-author

  1. Woolford, L., A. Rector, M. Van Ranst, A. Ducki, M. D. Bennett, P. K. Nicholls, K. S. Warren, R. A. Swan, G. E. Wilcox, and A. J. O’Hara (2007). 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. Journal of Virology. 81:13280-13290.
  2. Woolford, L., A. J. O’Hara, M. D. Bennett, M. Slaven, R. Swan, J. A. Friend, A. Ducki, C. Sims, S. Hill, P. K. Nicholls, and K. S. Warren (2008). Cutaneous papillomatosis and carcinomatosis in the western barred bandicoot (Perameles bougainville). Veterinary Pathology. 45(1):95-103.
  3. Woolford, L., M. Bennett, C. Sims, N. Thomas, J. Friend, P. Nicholls, K. Warren, and A. O’Hara (2009). Prevalence, emergence, and factors associated with a viral papillomatosis and carcinomatosis syndrome in wild, reintroduced, and captive western barred bandicoots (Perameles bougainville). EcoHealth.
  4. FitzGerald, L., M. Bennett, J. Ng, P. Nicholls, F. James, A. Eliott, U. Ryan. Morphological and molecular characterisation of a mixed Cryptosporidium muris/Cryptosporidium felis infection in a cat. [In preparation].

Scientific presentations

  • Parasitic infections of western barred bandicoots (Perameles bougainville)
  • Haematology and clinical chemistry of western barred bandicoots
  • Using in situ hybridization to detect genetic sequences of the putative Perameles bougainville papillomavirus type 1 in tissue biopsies
  • Isolation and attempted transmission and propagation of Perameles bougainville papillomavirus type 1

Western barred bandicoot symposium, Murdoch University, WA, February 2007.

  • Parasitic infections of western barred bandicoots (Perameles bougainville)

43rd International Symposium on Diseases of Zoo and Wild Animals, Edinburgh, UK, May 2007.

  • Papillomatosis in a southern brown bandicoot (Isoodon obesulus) in Western Australia

Wildlife Diseases Association – Australasian Chapter, Dryandra, WA, September 2007.

  • Papillomaviruses of marsupials

Invited speaker: Australian Society for Veterinary Pathology Conference, Fremantle, WA, October 2009.

Collaborators (positions held at July 2010) Dr. Lucy Woolford, Senior Clinical Training Scholar specialising in Anatomic Pathology at the Royal Veterinary College in London and the Animal Health Trust in Newmarket, England;
Professor Marc Van Ranst of the Laboratory of Clinical and Epidemiological Virology, Rega Institute for Medical Research, University of Leuven, Leuven, Belgium. 

Acknowledgements

This project is funded by the Australian Research Council in partnership with Murdoch University and the Western Australian Department of Conservation and Land Management (CALM – now DEC) through Linkage Project LP0455050. Our thanks go to Science Division CALM (DEC) officers for their work in establishing the western barred bandicoot captive breeding program. We are also grateful to June Butcher and the volunteers at Kanyana Wildlife Rehabilitation Centre for their care of the western barred bandicoots kept there; Dr. Jeff Short and the Useless Loop Community Biosphere Project Group for access to the Heirisson Prong western barred bandicoot population; Australian Customs and the crew of the ACV Corio Bay for assistance with fieldwork on Bernier and Dorre Islands; and the Narrogin CALM (DEC) officers, especially Howard Robinson, for help with fieldwork.

Photos provided by Mark Bennett

Department of Environment and ConservationAustralian Government | Australian Research CouncilKanyana Wild Life