Veterinary Science Collected Works - Theses
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ItemAn investigation into the role Toxoplasma gondii may play in the health of the southern brown bandicoot (Isoodon obesulus obesulus) and an assessment of environmental contamination with T. gondii( 2020)The southern brown bandicoot (Isoodon obesulus obesulus), a small, ground-dwelling marsupial, is listed as ‘endangered’ under the Australian Environment Protection and Biodiversity Conservation Act 1999. While many factors contributing to population decline are understood, in particular predation and loss of high-quality, connected habitat, there is a lack of knowledge about other threatening processes, including disease, which has contributed to declines in other small mammal species. Toxoplasma gondii, a protozoan intracellular parasite, excreted into the environment by cats, has been shown to cause clinical disease, including death, in many small and medium sized captive and free-ranging marsupials, including the eastern barred bandicoot (Perameles gunnii). Little is known of its effect on southern brown bandicoot populations. This study aimed to investigate the significance of T. gondii to the health of southern brown bandicoot populations on the northern hinterland of Western Port, Victoria and methods of predicting probability of infection with environmental T. gondii. A series of necropsies was performed on 33 southern brown bandicoots collected opportunistically over a five-year period. The causes of death were identified as motor vehicle trauma (22); predation (4); ejected pouch young (2); drowning (1); pyometra (1); possible toxicity and stomach bloating through lentil ingestion (1) and unknown (2). Real-time qPCR was performed on tissues from 30 cases, all of which were negative for the presence of T. gondii DNA. A range of helminths and ectoparasites were collected and identified, most of which had been previously been reported in this species. However, a metastrongyloid helminth species, found by histopathology in the lungs, is reported for the first time in a southern brown bandicoot. To assess environmental contamination with T. gondii, two sites with different cat densities were compared. Seroprevalence of antibodies to T. gondii (n=24) using the Modified Agglutination Technique was performed on trapped southern brown bandicoots. No evidence of infection with T. gondii was found at either site. Molecular (qPCR) methods were used to measure T. gondii oocyst presence in soil samples (n= 594) and prevalence of T. gondii in tissues of rabbits (n=118) and mice (n=267). All tests were negative across both sites except for the presence of T. gondii in one rabbit (prevalence 0.85%). These results suggest that rabbits and mice may have the potential to be reliable sentinel species and inform conservation management of the probability of infection with T. gondii in small marsupials. The results from this study are consistent with the concept that opportunistic necropsy is a valuable strategy for passive disease and cause of death surveillance in native wildlife. No evidence was found that T. gondii was impacting the health of southern brown bandicoots, however, further longitudinal health surveys are necessary to determine the true prevalence of disease and causes of mortality. Further studies are recommended to confirm the effectiveness of mice and rabbits as potential sentinel species in a range of ecosystems, including those environments which have higher levels of contamination with T. gondii.
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ItemMolecular epidemiological investigations of Enterocytozoon bieneusi( 2019)Enterocytozoon bieneusi is a microsporidian microorganism that causes intestinal disease in animals including humans. Although E. bieneusi has been discovered and investigated in numerous countries around the world, at the commencement of this PhD project, there was no record of E. bieneusi in animals in Australia and no information on the prevalence and genetic make-up of E. bieneusi populations in this country. Thus, the present project explored E. bieneusi of a number of species of wild and domestic animals as well as humans in parts of Australia using a molecular approach, in order to improve our knowledge of the epidemiology of this microsporidian. Nested-PCR-based sequencing of the internal transcribed spacer (ITS) and/or small subunit (SSU) of nuclear ribosomal DNA was used for the detection of E. bieneusi DNA in faecal samples and the genotypic characterisation of this microbe. In the first instance, three species of wild deer (Cervus elaphus, Dama dama and Rusa unicolor; n = 610) and three species of wild marsupials (Macropus giganteus, Vombatus ursinus and Wallabia bicolor; n = 1365) in Melbourne's water catchment areas were investigated (Chapters 2 and 3). Here, E. bieneusi was detected in 4.1% of sambar deer and 1.4% of marsupials. Phylogenetic analyses of sequence data showed that genotypes D, J, MWC_d1, MWC_d2 and Type IV (in sambar deer) and MWC_m1 and NCF2 (in marsupials) clustered with E. bieneusi genotypes recorded previously in humans, indicating the zoonotic potential of these genotypes. Studies of cattle on farms located near Tarago reservoir (n = 471; Chapter 4) in Victoria, and alpacas in the states of New South Wales, Queensland, South Australia, Victoria and Western Australia (n = 81; Chapter 5) revealed relatively high prevalences of E. bieneusi in both cattle (10.4%) and alpacas (9.9%). The phylogenetic analysis of ITS sequence data revealed six genotypes (BEB4, I and J, TAR_fc1, TAR_fc2 and TAR_fc3) in cattle and three (ALP1, APL3 and P) in alpacas, all recognised to have zoonotic potential, indicating that these herbivores might act as reservoirs for human infection. Subsequently, a study of companion animals (cats and dogs; n = 514) identified genotype D, which is commonly detected in humans (Chapter 6). However, surprisingly, an investigation of humans (n = 605) in the states of Queensland and Western Australia (Chapter 7) did not identify the genotypes previously found in cats, dogs and wildlife in the state of Victoria, although, surprisingly, a known genotype (i.e., ALP1) recorded in farmed alpacas (Chapter 5) was identified in humans for the first time (Chapter 7). This latter finding raises questions about the transmissibility of this genotype from alpaca to humans. Overall, this thesis has recorded E. bieneusi, for the first time, in a variety of animals in Australia, and provides a first glimpse of the epidemiology of this microsporidian in this country (Chapter 8). The findings of this thesis indicate that the animals studied here, including eastern grey kangaroos, swamp wallabies and wombats; sambar deer, cattle and alpacas; and cats and dogs, can all carry E. bieneusi genotypes that might infect humans via water, food and/or the environment. To provide further evidence to support this proposal and much deeper insights, well-designed large-scale (temporal and spatial) epidemiological studies are required. Population genomics of E. bieneusi using advanced next-generation sequencing and informatics tools could significantly support this endeavour. Other future investigations might also, for example, attempt to establish in vitro culture and in vivo systems for E. bieneusi for controlled studies of the biology of distinct genotypes of this microbe as well as approaches for assessing the viability and infectivity of E. bieneusi and anti-infectives against this microbe. Clearly, we are only beginning to understand some aspects of this enigmatic pathogen - E. bieneusi.