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Fauna translocations play an integral role in the management of threatened wildlife, though we are limited by our understanding of how the host-parasite community changes during translocation. During this longitudinal field-based study, we monitored gastrointestinal, blood-borne and ectoparasite taxa infecting woylies (Bettongia penicillata) for up to 12 months following two fauna translocations to supplement existing wild woylie populations in three different sites (Dryandra, Walcott and Warrup East) within the south-west of Western Australia. We aimed to (a) identify changes in parasite community structure of both translocated and resident woylies following translocation; and (b) evaluate the efficacy of ivermectin treatment in translocated hosts. Destination site and time since translocation had the strongest effects on parasite prevalence and mean faecal egg counts following translocation. Ivermectin treatment did not significantly reduce parasite prevalence or mean faecal egg counts in treated hosts. Prior to translocation, parasite community composition differed significantly between woylies selected for translocation and resident woylies within each release site. Following translocation, the parasite communities of translocated and resident hosts converged to become more similar over time, with loss of parasite taxa and novel host-parasite associations emerging. This is the first study to examine changes to the broader parasite community in translocated and resident animals following translocation. The dominant site-specific response of parasites following translocation reinforces the importance of incorporating parasite studies to enhance our fundamental understanding of perturbations in host-parasite systems during translocation, in particular the site-level drivers of parasite dynamics. Keywords: Bettongia penicillata, Host-parasite relationship, Ivermectin, Polyparasitism, Wildlife, Woylie

Abstract Background Understanding how fauna translocation and antiparasitic drug treatment impact parasite community structure within a host is vital for optimising translocation outcomes. Trypanosoma spp. and piroplasms (Babesia and Theileria spp.) are known to infect Australian marsupials, including the woylie (Bettongia penicillata). However relatively little is known about these haemoparasites, or how they respond to management practices such as translocation. We monitored haemoparasites infecting woylies for up to 12 months during two fauna translocations to supplement existing woylie populations in three different sites (Dryandra, Walcott and Warrup East) within south-western Australia between 2014 and 2016, with the aim of investigating (i) how haemoparasite prevalence, Trypanosoma spp. richness and Trypanosoma spp. community composition varied over time and between different sites following translocation; and (ii) whether ivermectin treatment indirectly impacts haemoparasite prevalence. Using molecular methods, 1211 blood samples were screened for the presence of trypanosomes, and a subset of these samples (n = 264) were also tested for piroplasms. Results Trypanosomes and piroplasms were identified in 55% and 94% of blood samples, respectively. We identified five Trypanosoma species, two Theileria species, a single species of Babesia and a novel Bodo species. Trypanosoma spp. richness and the prevalence of haemoparasite co-infection increased after translocation. Prior to translocation, Trypanosoma spp. community composition differed significantly between translocated and resident woylies within Walcott and Warrup East, but not Dryandra. Six months later, there was a significant difference between translocated and resident woylies within Dryandra, but not Walcott or Warrup East. The response of haemoparasites to translocation was highly site-specific, with predominant changes to the haemoparasite community in translocated woylies occurring within the first few months following translocation. Ivermectin treatment had no significant effect on haemoparasite prevalence. Conclusions This study contributes to our understanding of haemoparasite dynamics in woylies following translocation. The highly site-specific and rapid response of haemoparasites to translocation highlights the need to better understand what drives these effects. Given that haemoparasite prevalence and composition of translocated and resident animals changed significantly following translocation, we propose that parasite monitoring should form an essential component of translocation protocols, and such protocols should endeavour to monitor translocated hosts and cohabiting species.

During monitoring of critically endangered woylie (Bettongia penicillata) populations within the south-west of Western Australia, an adult female woylie was euthanased after being found in extremely poor body condition with diffuse alopecia, debilitating skin lesions and severe ectoparasite infestation. Trypanosoma copemani G2 and Sarcocystis sp. were detected molecularly within tissue samples collected post-mortem. Potorostrongylus woyliei and Paraustrostrongylus sp. nematodes were present within the stomach and small intestine, respectively. Blood collected ante-mortem revealed the presence of moderate hypomagnesaemia, mild hypokalaemia, mild hyperglobulinaemia and mild hypoalbuminaemia. Diffuse megakaryocytic hypoplasia was evident within the bone marrow. We propose various hypotheses that may explain the presence of severe ectoparasite infection, skin disease and poor body condition in this woylie. Given the potential deleterious effects of parasite infection, the importance of monitoring parasites cannot be over-emphasised. Keywords: Bettongia penicillata, Fauna translocation, Paraustrostrongylus sp., Polyparasitism, Sarcocystis sp., Trypanosoma copemani

Vector-borne haemoprotozoans comprise a diverse group of eukaryote single-celled organisms transmitted by haematophagous (blood-feeding) invertebrates. They can cause debilitating diseases that impact wildlife, livestock, companion animals and humans. Recent research has shown that Australian wildlife host a diverse range of haemoprotozoan species; however, to date this work has primarily been confined to a few host species or isolated populations in rural habitats. There has been little investigation into the presence of these blood parasites in wildlife inhabiting urban and peri-urban areas. In this study, blood and tissue samples and ticks were collected from wildlife in New South Wales and Western Australia. Extracted DNA samples were screened with pan-specific molecular assays to determine the presence of haemoprotozoans using amplicon metabarcoding and Sanger sequencing approaches. In addition, light microscopy was performed on blood films. Eight haemoprotozoans were identified in the present study, which included species of Babesia, Hepatozoon, Theileria and Trypanosoma. Blood samples were collected from 134 animals; 70 black rats (Rattus), 18 common brush-tailed possums (Trichosurus vulpecula), two bush rats (Rattus fuscipes), 22 chuditch (Dasyurus geoffroii), 20 long-nosed bandicoots (Perameles nasuta), one quenda (Isoodon fusciventer) and one swamp rat (Rattus lutreolus). Molecular screening of DNA extracted from blood samples identified 52.2% (95% CI: 43.8–60.5%) of individuals were positive for at least one haemoprotozoan species, with 19.4% (95% CI: 13.4–26.7%) positive for more than one species. The present study provides the first sequences of Theileria cf. peramelis from black rats and long-nosed bandicoots. Babesia lohae was identified from brush-tailed possums. Two Hepatozoon genotypes were identified from black rats and bush rats. Black rats showed the highest haemoprotozoan diversity, with five species identified. No known human pathogens that have been described in the northern hemisphere were identified in the present study, and future work is required to understand the zoonotic potential of these microbes in Australia. This work represents the first large-scale body of research using molecular tools to investigate haemoprotozoans in animals at the urban-wildland interface. Further research is needed to investigate potential consequences of infection in wildlife, particularly effects of pathogen spillover from invasive black rats to native wildlife.